Naomi Lubick
Posted: 05/23/2011; Environmental Health Perspectives. 2011;119(5):a214-a217. © 2011 National Institute of Environmental Health Sciences
Abstract and Introduction
Introduction
When a team of researchers from Sweden first started measuring chemicals in a river near Patancheru, India, they found shocking concentrations of drugs flowing downstream—for example, levels of the potent antibiotic ciprofloxacin greater than those found in the blood of humans taking the drug. A major source of these drugs was treated wastewater from pharmaceutical manufacturing plants that was discharged into the river and surrounding environs, as Joakim Larsson and his colleagues from the University of Gothenburg reported several years ago.[1] An update published in PLoS ONE [2] now links the drugs with downstream development of microbes with genetic resistance to multiple antibiotics typically used to treat human illness.
The researchers found snippets of genetic material in bacteria from river sediments downstream of the treatment plant that conferred resistance not only to ciprofloxacin, a fluoroquinolone, but also to betalactams, aminoglycosides, sulfonamides, and other classes of antibiotics. Several genes that provide resistance to ciprofloxacin and have the ability to transfer between different bacteria were extremely common at some of the sampling sites.[2]
What if the bacteria in Patancheru could develop ways to survive the daily onslaught of ciprofloxacin, most likely over the course of years in their river environment, and ended up passing on their new genetic resistance to pathogenic bacteria that could be a threat to human health? Although Larsson's team has yet to catalog antibiotic resistance in the local population, people in the region are continually exposed to resistant microbes as they use the river water for agriculture and everyday home life. "This is a huge scary experiment in nature," Larsson says.
Just how isolated these kinds of drug "hot spots" are remains unknown, although researchers have pressed for global monitoring of antibiotic use and resistance for the past several decades, across disciplines as diverse as clinical medicine and ecotoxicity. Bringing together these fields reflects the breadth of challenges in tracking antibiotic resistance, but new technologies and ideas hold promise for the near future.
Overcoming a Lack of Coordination
"Misuse of antibiotics is obviously what creates the basic factors that produce drug resistance," says Mario Raviglione, director of the World Health Organization (WHO) department charged with tuberculosis control; this is true in both the developing and developed worlds. And despite educational campaigns by the U.S. Centers for Disease Control and Prevention (CDC)[3] and others aimed at improving clinicians' use of antibiotics, overprescribing remains a problem for multiple reasons.[4] Moreover, patient compliance—for example, taking the full course of prescribed antibiotics—can be lax, which leads to the evolution of more antibiotic-resistant pathogens.
Agricultural use of human drugs adds to the threat of drug resistance. After World War II, antibiotics started to be used for purposes such as growth promotion in livestock. Since then, antibiotics—and in some cases, the genes for resistance to multiple drugs—have been found on industrial cattle, swine, and shrimp farms,[5,6,7,8] measured on chicken skins in grocery stores,[9] and even detected in apple orchards sprayed with drugs originally intended for human use.[10]
For World Health Day in April 2011 the WHO chose the theme of the global spread of antibiotic resistance, marking a little over a decade since the organization first called for patient and doctor guidelines to protect antibiotics from becoming obsolete.[11] A document issued by the WHO in 2001 put forth a series of recommendations for patients and the general community, prescribers and dispensers, hospitals, agricultural enterprises, national governments and health systems, and drug developers and promoters.[12] However, in general "very few countries, if any, have made a comprehensive effort to do any of the measures included in the older guidelines," says Raviglione, who led preparations for World Health Day 2011. "Why are countries not picking them up? Lack of resources? Their health systems are not strong enough? The cost of drugs?"
On 7 April 2011 the organization released updated policy guidance for countries to curb the spread of antibiotic resistance in health care settings.[13] Some of this guidance is aimed at doctors and hospitals, while some is geared toward policy makers and legislators. The simple package of policy recommendations is intended to be easy for countries to adopt, Raviglione says. The WHO-level focus on antibiotic resistance issues also gives health ministers around the world a platform from which to call for funding and research attention at home.
But individual countries cannot handle this issue acting alone. During its 2009 presidency of the European Union, the Swedish government highlighted antibiotic resistance, focusing the conversation on solutions across Europe.[14] For instance, a September 2009 meeting targeted industry, policy makers, and others focused on finding incentives for creating new drugs.[15] Meanwhile, in the United States, the U.S. Food and Drug Administration (FDA) in 2010 issued draft guidance urging the judicious use of medically important antibiotics in livestock.[16] The 2011–2015 Strategic Plan of the National Antimicrobial Resistance Monitoring System, a collaboration between the FDA, CDC, U.S. Department of Agriculture, and state and local health departments, includes efforts to strengthen sampling, reporting, and international and domestic collaborative efforts.[17]
Bills aimed at addressing antibiotic resistance were introduced in 2009 in the House of Representatives[18] and the Senate,[19] but foundered. This year, Louise Slaughter (D–NY) tried again in the House, introducing H.R. 965, the Preservation of Antibiotics for Medical Treatment Act of 2011[20] on March 9. Another bill in the House, H.R. 6331, the Generating Antibiotic Incentives Now Act of 2010,[21] would create incentives to bring new antibiotics to market by speeding up the approval process. These bills have lingered in committee, even as the problem continues to grow globally.
Traveling Wild
The antibiotic drug hot spot in India is not alone: researchers have traced drugs flowing downstream from manufacturers in China and Cuba[22] and from wastewater treatment plants in the United States.[23,24] They also have identified antimicrobial resistance genetic material in treated waste effluent and tap water in Michigan and Ohio,[25,26] and researchers in Sweden recently documented multidrug-resistant Escherichia coli in the waste of migrating birds in the Arctic.[27]
Given the widespread presence of antibiotics in the natural environment,[28] it should not be a surprise that resistance is growing in wild bacteria and microbes—but is that resistance transferable to pathogens relevant to human health? Eventually the answer will most certainly be yes: Bacteria, microbes, and even fungi under stress from high concentrations of drugs might have the ability to replicate DNA snippets and possibly pass them on to other microbial species in the environment, says Dave Ussery, an associate professor of microbial genomics at the Technical University of Denmark. Transported in the integrons, plasmids, and other cellular genetic modules that confer resistance, these snippets can be traded like baseball cards among microbes.[29]
The locations and impacts of reservoirs of antibiotic resistance in the wild remain enigmatic,[30] and not only for humans. Antibiotic drugs and any acquired resistance to them might, for example, affect how microbes communicate with each other through "quorum sensing," a protein signaling feedback loop key to microbial population dynamics.[31]
But until recently, truly wild settings have garnered less interest from researchers, policy makers, and media than agricultural exposures—for example, farmers who handle pigs treated with multiple drugs and then end up with resistant strains on their skin,[32] including methicillin-resistant Staphylococcus aureus.
Researchers from the U.S. Geological Survey led by Dana Kolpin also are looking at animal waste for how much residual concentration of antibiotic it might carry and the potential impacts on microbial life if the manure is spread on agricultural fields or released accidentally. Results of Kolpin's research are not yet available, but a small-scale study by another group tracking resistance genes from biosolids and manure at two soil sites has shown the genes—with resistance for tetracycline and sulfanomides—transfer at different rates depending on soil type.[33]
Kink in the Pipeline
Another worry underlying the issue of resistance is the fact that pharmaceutical companies are not discovering new antibiotics. At least three reasons explain why the pipeline is so empty, says Ingrid Petersson, director of science relations at pharmaceutical company AstraZeneca. First, finding new pathways in microbes or pinpointing proteins to create new antibiotics is difficult, in part because of what could be considered an embarrassment of possibilities with too many unknowns. Second, she says, the regulatory environment is complicated: getting a drug through approval processes takes a long time and costs a lot of money, among other factors. And third, low prices for existing antibiotics—many of which are generics—do not encourage companies to invest in new drugs.
"Existing, older antibiotics are cheaper, which makes it difficult to achieve realistic prices for new antibiotics—prices which would provide a viable return on investment," explains Colin Mackay, director of communication and partnerships for the European Federation of the Pharmaceutical Industries and Associations, a trade organization. "Furthermore, antibiotics are only used acutely, perhaps only for a week to ten days at a time. This adds to the difficulty in making a return on investment. By comparison, treatments for chronic illnesses, say for cardiovascular or musculoskeletal conditions, are used long term, perhaps for the rest of the patient's life."
"Drug development … is one of the most critical things that we are facing," says Otto Cars, the chairman of ReAct (Action on Antibiotic Resistance), an independent think tank, and a professor of infectious diseases at Uppsala University. For gut flora that can shift readily from animal to human hosts—including Gram-negative[34] enterics such as Salmonella, Campylobacter, Klebsiella, E. coli, and Shigella—horizontal gene transfer is "moving rapidly now," he says. "These kinds of infections that Gram-negative bacteria are causing are already untreatable; even in the rich part of the world, there are totally resistant strains," he says. "The drug pipeline is particularly empty for that space."
Nevertheless, some major companies are looking into new antibiotics. For example, AstraZeneca is looking for solutions for multidrug-resistant tuberculosis.[35,36] Companies are investing money, sometimes by acquiring smaller companies that have begun the research or joining in efforts with nonprofits[37] and academic researchers. Petersson notes that the Trans Atlantic Task Force on Antimicrobial Resistance,[38] formed between the European Union and the United States as part of the 2009 EU–US Summit Declaration,[39] will present suggestions at this year's summit meeting for areas of cooperation, including incentives for industry to pursue new drug development.
The WHO, the CDC, and other national, international, and nonprofit organizations are pursuing alternative business models. Government funding, as when a federal agency invests in vaccine research, may be one option, Cars suggests. So-called advanced market commitments—where governments pledge to purchase drugs, thus guaranteeing a market—is another option. In its new policy guidance,[13] the WHO calls for global and national commitments to develop drugs and share information on the national costs of inaction, as well as "push" and "pull" incentives to reduce the inherent risks in the initial phases of research and development and to offset the risks of an uncertain market, respectively.
Crossing Boundaries
Heeding calls[40-42] for better management of the drugs currently available to doctors will require much more attention to trends of resistance. In particular, monitoring is now lacking. "If anything, we don't know enough about developing countries to understand the situation—what resistant bacteria are there? In Europe and the U.S., systems of surveillance are in place, but not in most of Africa or Asia," Raviglione says, referring to health systems, although the same holds true for environmental surveys.
Europe and the United States use far more antibiotics by volume as well as newer antibiotics compared with less affluent countries that typically use fewer and older generations of drugs, Raviglione says. That would indicate such countries might not yet have resistance to latest-generation drugs the same way Europe and the United States do. But there remains concern about the outsourcing of drug production to developing countries, especially India and China, where lax enforcement of regulations could increase the likelihood of unchecked environmental releases of active pharmaceutical ingredients—hence studies such as Larsson's work in Patancheru. The potential for impacts of manufacturing newer antibiotics in developing countries, with possible unwanted environmental releases, has not yet been studied, say the scientists contacted for this story.
Making data internationally available so that teams are tracking the same genes and species in different countries may be one avenue of attack on the issue of antibiotic resistance. Julian Davies, a professor emeritus of microbiology and immunology at the University of British Columbia, and David Graham, an environmental engineer at Newcastle University, have been working on a proposal to bring together members of the medical, environmental, and microbial research communities to address antibiotic resistance issues. If funded, their efforts could result in local centers on every continent working to monitor antibiotic resistance, look for simple solutions to preventing the spread of antibiotic-resistant microbes inside hospitals, and communicate about antibiotic resistance issues at an international scale.
The interdisciplinary breadth needed to address the many issues at hand have led to miscommunications stemming from vocabulary, Graham says. Take the word transmission, he explains: "To a physician or engineer, transmission means migration between individuals at larger scales, whereas to a microbiologist transmission is something at the micro-scale between the bacteria themselves. It took us [the diverse members of the team working on the proposal] awhile to agree upon a common language."
But microbes have few such communication barriers, and they quickly find ways to communicate resistance, as plane transit brings countries—and antibiotic resistance hot spots—closer together. Some of the global aspects of the problem can be illustrated by last year's description of NDM-1, a protein present on plasmids that confers antibiotic resistance to multiple antibiotics.[43] The mechanism of resistance traveled from hospitals in India to the United Kingdom via patients who had visited the subcontinent, presumably for cheap medical treatments, and then returned home. Furthermore, NDM-1 has appeared in tap water and wastewater outside of hospital settings in New Delhi, according to another recent report in The Lancet Infectious Diseases,[44] heightening concerns for local transmission in an urban environment.
Tools for the Trade
Ussery and his colleagues, while working to figure out antibiotic resistance transfer at a basic level, are also developing field tools for tracking antibiotic resistance genes and the microbes that carry them. The team includes computer scientists searching for simple algorithms that will let researchers make fast identifications of resistance gene sequences and resistant microbial species.
Ussery envisions a device on every doctor's desk that could take a swab from a patient and sequence the DNA from that sample. This device, connected to the Internet or with databases preloaded, could use the sequence to identify the microbes present to the genus or even species level, then spot genes they might carry for resistance to certain drugs. The test could potentially even predict the effectiveness of specific drugs in individual patients. That could guide doctors in prescribing treatment or hospitals in determining when to quarantine a patient with a particularly virulent resistant strain of an infectious disease.
"Some machines are now doing single-molecule sequencing [of bacteria] from one cell. The technology is almost there," Ussery says, citing manufacturer Oxford Nanopore's machines that can sequence a microbe from one cell as being close to market-ready.
For tracking antibiotic resistance in the field, these techniques could prove to be a massive boon, but they will have to become much less expensive than current market costs of the machines, comments Davies. The newest so-called third-generation apparatuses for identifying genetic codes in the blink of an eye may cut the cost of reading an entire genome to a tenth or less of current costs, but machines will still cost hundreds of thousands of dollars, he says, backed up by the price tag of Pacific Biosciences' first entry to the market late last year.
Searching for Simple Answers
Even a very expensive diagnostic tool would save a lot of lives, Cars says, and it would also save antibiotics for clinical use: rapid tests used on a daily basis could lead to hospital practices that save time and money while conserving antibiotics for treatment.
But new technologies are not the only tool that will be necessary to address all the threads that intertwine the problem of spreading antibiotic resistance, Cars and others say. Tracking genes in hospitals and the wild, putting policies in place to limit use of antibiotics outside of absolute necessity in a clinical setting—the wish list goes on.
And yet, in some ways, simple solutions seem to be in the offing. U.S. congressmen recently visited Denmark to learn more about that country's successful transition away from antibiotic use for growth promotion in swine.Contrary to stories about how Denmark's agricultural sector crashed after that use of antibiotics completely stopped five years ago, Frank Møller Aarestrup of the Technical University of Denmark and colleagues report the country today continues to increase its exports of pigs without the help of antibiotics, depending solely on improved animal husbandry techniques.[45]
This anecdote underscores the power of seemingly simple policy decisions to make immediate changes with promising consequences. Absent such courageous steps, "it's almost just a matter of time" before antibiotic resistance is transferred to a pathogen that matters for human health, Ussery says.
"It's an enormously frustrating situation," says Davies, who says he feels he and his colleagues talk a lot about the problem without making much headway at the larger scale. In the end, he says, solutions can only come from convincing the general public and lawmakers that the time to act is now.
References
1.Larsson DGJ, et al. Effluent from drug manufactures contains extremely high levels of pharmaceuticals. J Hazard Mat 148(3):751–755 (2007); doi:10.1016/j.jhazmat.2007.07.008.
2.Kristiansson E, et al. Pyrosequencing of antibiotic-contaminated river sediments reveals high levels of resistance and gene transfer elements. PLoS ONE 6(2):e17038 (2011); doi:10.1371/journal.pone.0017038.
3.CDC. Get Smart for Healthcare [website]. Atlanta, GA:U.S. Centers for Disease Control and Prevention (updated 25 Mar 2011). Available: http://tinyurl.com/6jyh49q [accessed 6 Apr 2011].
4.Kuehlein T, et al. Antibiotic prescribing in general practice—the rhythm of the week: a cross-sectional study. J Antimicrob Chemother 65(12):2666–2668 (2010); doi: 10.1093/jac/dkq364.
5.Smith TC, et al. Methicillin-resistant Staphylococcus aureus (MRSA) Strain ST398 is present in Midwestern U.S. swine and swine workers. PLoS ONE 4(1):e4258; doi:10.1371/journal.pone.0004258.
6.Schmidt CW. Swine CAFOs & novel H1N1 flu: separating facts from fears. Environ Health Perspect 117(9):A394–A401 (2009); doi:10.1289/ehp.117–a394.
7.Chapin A, et al. Airborne multidrug-resistant bacteria isolated from a concentrated swine feeding operation. Environ Health Perspect 113(2):137–142; doi:10.1289/ehp.7473.
8.Xuan Le T, et al, Antibiotic resistance in bacteria from shrimp farming in mangrove areas. Sci Total Environ 349(1–3):95–105 (2005); doi:10.1016/j.scitotenv.2005.01.006.
9.Vincent C, et al. Food reservoir for Escherichia coli causing urinary tract infections. Emerg Infect Dis 16(1):88–95 (2010); doi:10.3201/eid1601.091118.
10.Re: U.S. EPA Specific Exemption to the Michigan Department of Agriculture for the Use of Gentamicin, Formulated as the Unregistered Product Agry-Gent 10W, on Apples to Control Fire Blight. Letter from Lois Rossi to Brian Verhougstraete, 24 April 2008. Available: http://tinyurl.com/3e8pcw4 [accessed 6 Apr 2011].
11.WHO. World Health Organization Report on Infectious Diseases 2000. Overcoming Antimicrobial Resistance. A Message From the Director-General, World Health Organization. Geneva, Switzerland:World Health Organization (2000). Available: http://tinyurl.com/69ugd4k [accessed 6 Apr 2011],
12.WHO. WHO Global Strategy for Containment of Antimicrobial Resistance. Executive Summary. Geneva, Switzerland:World Health Organization (2001). Available: http://tinyurl.com/42xdzdx [accessed 6 Apr 2011].
13.WHO. World Health Day 2011: Policy Briefs [website]. Geneva, Switzerland:World Health Organization (2011). Available: http://tinyurl.com/3oypba8 [accessed 6 Apr 2011].
14.Cars O. Antibiotic Resistance Is a Global Public Health Threat That Can Only Be Solved By a World Working Together. Health–EU Newsletter. Brussels, Belgium:Community Public Health Programme, European Union (19 Nov 2009). Available: http://tinyurl.com/44nnyrw [accessed 6 Apr 2011].
15.The Swedish National Board of Health and Welfare. Antimicrobial Resistance—Inspiration and Exchange of Experience from EU Presidencies. Stockholm, Sweden:The Swedish National Board of Health and Welfare (Socialstyrelsen) (2010). Available: http://tinyurl.com/3bbmu5w [accessed 6 Apr 2011].
16.FDA. Draft Guidance #209. The Judicious Use of Medically Important Antimicrobial Drugs in Food-Producing Animals. Rockville, MD:Center for Veterinary Medicine, Food and Drug Administration, U.S. Department of Health and Human Services (28 June 2010). Available: http://tinyurl.com/352ks4e [accessed 6 Apr 2011].
17.FDA. The National Antimicrobial Resistance Monitoring System (NARMS) Strategic Plan 2011–2015. FDA-2010-N-0620. Silver Spring, MD:U.S. Food and Drug Administration, U.S. Department of Health and Human Services (updated 25 Mar 2011).
18.H.R. 1549. Preservation of Antibiotics for Human Treatment Act of 2009. Available: http://tinyurl.com/dm8by3 [accessed 6 Apr 2011].
19.S. 619. Preservation of Antibiotics for Medical Treatment Act of 2009. Available: http://tinyurl.com/4yc2852 [accessed 6 Apr 2011].
20.H.R. 965. Preservation of Antibiotics for Human Treatment Act of 2011. Available: http://tinyurl.com/6hlwzkg [accessed 6 Apr 2011].
21.H.R. 6331. Generating Antibiotic Incentives Now (GAIN) Act of 2010. Available: http://tinyurl.com/3cqfavy [accessed 6 Apr 2011].
22.Graham DW, et al. Antibiotic resistance gene abundances associated with waste discharges to the Almendares River near Havana, Cuba. Environ Sci Technol 45(2):418–424 (2011); doi:10.1021/es102473z.
23.Phillips PJ, et al. Pharmaceutical formulation facilities as sources of opioids and other pharmaceuticals to wastewater treatment plant effluents. Environ Sci Technol 44(13):4910–4916 (2010); doi:10.1021/es100356f.
24.USGS. Manufacturing Facilities Release Pharmaceuticals to the Environment [website]. Washington, DC:Toxic Substances Hydrology Program, U.S. Geological Survey, U.S. Department of the Interior (updated 12 Nov 2010). Available: http://tinyurl.com/3kxjg8x [accessed 6 Apr 2011].
25.Xi C, et al. Prevalence of antibiotic resistance in drinking water treatment and distribution systems. Appl Environ Microbiol 75(17):5714–5718 (2009); doi:10.1128/AEM.00382-09.
26.Zhang Y, et al. Wastewater treatment contributes to selective increase of antibiotic resistance among Acinetobacter spp. Sci Total Environ 407(12):3702–3706 (2009); doi:10.1016/j.scitotenv.2009.02.013.
27.Sjölund M, et al. Dissemination of multidrug-resistant bacteria into the Arctic. Emerg Infect Dis 14(1):70–72 (2008 ); doi:10.3201/eid1401.070704.
28.Rosenblatt-Farrell N. The landscape of antibiotic resistance. Environ Health Perspect 117(6):A244–A250 (2009); doi:10.1289/ehp.117-a244.
29.Aarestrup FM, ed. Antimicrobial Resistance in Bacteria of Animal Origin. Washington, DC:ASM Press (2006).
30.Allen HK, et al. Call of the wild: antibiotic resistance genes in natural environments. Nature Rev Microbiol 8(4):251–259 (2010); doi:10.1038/nrmicro2312.
31.Yim G, et al. The truth about antibiotics. Int J Med Microbiol 296(2–3)163–170 (2006); doi:10.1016/j.ijmm.2006.01.039.
32.Silbergeld EK, et al. Industrial food animal production, antimicrobial resistance, and human health. Annu Rev Public Health 29:151–169 (2008); doi:10.1146/annurev.publhealth.29.020907.090904.
33.Munir M, Xagoraraki I. Levels of antibiotic resistance genes in manure, biosolids, and fertilized soil. J Environ Qual 40(1):248–255 (2011); doi:10.2134/jeq2010.0209.
34.Gram-negative bacteria, with their thinner cell walls, tend to be more likely to develop resistance compared with thicker-walled Gram-positive bacteria.
35.AstraZeneca. Dedicated TB Research [website]. Södertälje, Sweden:AstraZeneca Global (2011). Available: http://tinyurl.com/679gx84 [accessed 6 Apr 2011].
36.AstraZeneca. Bacterial Resistance to Antibiotics is a Global Health Threat. Creative Collaboration is the Key to Successfully Meeting the Challenge [website]. Södertälje, Sweden:AstraZeneca Global (updated 1 Nov 2010). Available: http://tinyurl.com/3hr9tp4 [accessed 6 Apr 2011].
37.Resources for the Future Launches Global Antibiotic Resistance Initiative [press release]. Seattle, WA:Bill & Melinda Gates Foundation (7 Jan 2009). Available: http://tinyurl.com/6cvood4 [accessed 6 Apr 2011].
38.ECDC. Trans Atlantic Task Force on Antimicrobial Resistance—TATFAR [website]. Stockholm, Sweden:European Centre for Disease Prevention and Control (2005-2011). Available: http://tinyurl.com/5sdxlpm [accessed 6 Apr 2011].
39.Swedish Presidency of the European Union [website]. 2009 EU-U.S. Summit Declaration. Stockholm, Sweden:Communications Secretariat for Sweden's EU Presidency, Prime Minister's Office of Sweden (2009). Available: http://tinyurl.com/3u9ndv5 [accessed 6 Apr 2011].
40.ECDC. European Antibiotic Awareness Day [website]. Stockholm, Sweden:European Centre for Disease Prevention and Control (2005-2011). Available: http://tinyurl.com/5to66ko [accessed 6 Apr 2011].
41.ReAct. Antibiotic Awareness Day—Not Only in Europe [press release]. Uppsala, Sweden:ReAct, Uppsala University (22 Dec 2010). Available: http://tinyurl.com/6zmjau9 [accessed 6 Apr 2011].
42.ECDC. ECDC Marks World Health Day 2011 with Situation Update on Antimicrobial Resistance in Europe [press release]. Stockholm, Sweden:European Centre for Disease Prevention and Control (19 Apr 2011). Available: http://tinyurl.com/ykpjtlz [accessed 19 Apr 2011].
43.Kumarasamy KK, et al. Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study. Lancet Infect Dis 10(9):597–602 (2010); doi:10.1016/S1473-3099(10)70143-2.
44.Walsh TR, et al. Dissemination of NDM-1 positive bacteria in the New Delhi environment and its implications for human health: an environmental point prevalence study. Lancet Infect Dis; doi:10.1016/S1473-3099(11)70059-7 [online 6 Apr 2011].
45.Aarestrup FM, et al. Changes in the use of antimicrobials and the effects on productivity of swine farms in Denmark. J Am Vet Med Assoc 71(7):726–733 (2010); doi:10.2460/ajvr.71.7.726.
jueves, 9 de junio de 2011
Be on Alert for 'Super Toxic' Bug in Travelers, CDC Says
E coli Infection 'Very Rare' but Deadly
Nancy A. Melville
June 3, 2011 — As health officials in Germany continued to seek the source of a uniquely toxic enterohemorrhagic Escherichia coli outbreak that has claimed the lives of at least 18 people, the Centers for Disease Control and Prevention (CDC) issued a notice to healthcare providers to be on alert for the Shiga toxin–producing E coli O104:H4 (STEC O104:H4) infections among travelers returning from Germany.
While there are some reports of the outbreak stabilizing, the World Health Organization (WHO) confirms that a total of 1823 cases of STEC O104:H4 have been reported, including 520 cases of hemolytic uremic syndrome (HUS), a potentially life-threatening complication of the infection that can cause kidney failure. Twelve HUS cases were fatal, and 6 deaths were reported among non-HUS cases.
The number of countries reporting cases of the STEC O104:H4 poisoning had increased to 11 on Friday. However, all but 1 of the deaths since the outbreak emerged in May have occurred in Germany. The 18th death was reportedly in Sweden and involved a person who had recently returned from Germany.
Symptoms of the strain, which European authorities have called "super-toxic," are notably severe, including stomach cramps, bloody diarrhea, vomiting, and fever. However, fever is not usually high.
Rare but Not Unfamiliar
Four suspected cases of the infection have been reported in the United States, all involving people who had recently traveled to Hamburg, Germany, said Chris Braden, MD, Director of Foodborne, Waterborne, and Environmental Diseases for the CDC, at a press briefing today.
Three of the 4 cases in the United States involved HUS and the patients were hospitalized. "The fourth case did not develop HUS but had bloody diarrhea, and we know there was a Shiga toxin–producing organism involved," he said.
Dr. Braden described the STEC O104:H4 strain as "very rare" but not entirely unfamiliar.
"The CDC is not aware of any confirmed cases of this infection ever reported in the US. However, we have become aware of similar strains in other countries in the past," he said.
The strain attacks the body in a manner unlike other strains of Shiga-producing E coli.
"The strain is different in its genetic markers and in the way it attaches to the lining of the intestine," Dr. Braden added.
Most Victims Female
In addition to causing particularly severe symptoms, the strain is unusual in that most of its victims appear to be women and people over the age of 20.
"It is true that in Germany 60% of the enterohemorrhagic E coli cases and 71% of the HUS cases are female," the WHO confirmed.
The unusual patterns underscore that little is known about the strain's unusual nature, but they could also suggest the source that may be somehow related to the adult female demographic, Dr. Braden said.
"We have a lot to learn about this particular organism. It's possible this organism had a predilection for adults over children, but it's also possible the type of food or produce is being eaten [more] by adult women than others."
Dr. Braden also noted that the duration time from exposure to onset of symptoms is also unique, with incubation times of more than a week to up to 12 days, compared to as little as 5 days commonly seen with other E coli infections.
Most patients' symptoms resolve within 5 to 7 days. However, HUS can develop a week after diarrhea begins.
"The classic triad of findings in HUS is acute renal damage, microangiopathic hemolytic anemia (evidence of schistocytes and helmet cells on peripheral blood smear), and thrombocytopenia," the CDC explained in a statement.
Prime Suspects
After backtracking on an earlier suggestion that the source of the E coli strain could be linked to organic Spanish cucumbers, German officials maintain that cucumbers, tomatoes, and lettuce are top suspects.
In addition, the Robert Koch Institute, Germany's national disease control agency, has advised consumers, particularly those in the northern Germany region around Hamburg, to avoid those vegetables.
The Institute has said that the number of cases appeared to have peaked around May 21 or 22. However, officials cautioned that communication delays may have slowed the reporting of new cases since then. In the meantime, German officials are not letting their guard down.
"We are dealing here in fact with the biggest epidemic caused by bacteria in recent decades," Reinhard Brunkhorst, president of the German Nephrology Society, told reporters in Hamburg this week.
In treating suspected STEC cases, some research has shown that administering antibiotics may in fact increase their risk of developing HUS, but the CDC recommends that clinicians ultimately determine treatment according to each individual patient.
"There may be indications for antibiotics in patients with severe intestinal inflammation if perforation is of concern," the agency said. "Of note, isolates of STEC O104:H4 from patients in Germany have demonstrated resistance to multiple antibiotics."
Detection Guidelines
The agency issued the following additional guidelines for detecting and characterizing STEC infections:
•All stools submitted for testing from patients with acute community-acquired diarrhea should be cultured for STEC O157:H7. These stools should be simultaneously assayed for non-O157 STEC with a test that detects the Shiga toxins or the genes encoding these toxins.
•Clinical laboratories should report and send E coli O157:H7 isolates and Shiga toxin–positive samples to state or local public health laboratories as soon as possible for additional characterization.
•Specimens or enrichment broths in which Shiga toxin or STEC are detected, but from which O157:H7 STEC isolates are not recovered, should be forwarded as soon as possible to a state or local public health laboratory so that non-O157:H7 STEC can be isolated.
•It is often difficult to isolate STEC in stool by the time a patient presents with HUS. Immunomagnetic separation (IMS) has been shown to increase recovery of STEC from HUS patients. For any patient with HUS without a culture-confirmed STEC infection, stool can be sent to a public health laboratory that performs IMS or to the CDC (through a state public health laboratory). In addition, serum can be sent to CDC (through a state public health laboratory) for serologic testing of common STEC serogroups.
martes, 10 de mayo de 2011
The 10 Most Prescribed Drugs
Daniel J. DeNoon
April 20, 2011 — The 10 most prescribed drugs in the U.S. aren't the drugs on which we spend the most, according to a report from the IMS Institute for Healthcare Informatics.
The institute is the public face of IMS, a pharmaceutical market intelligence firm. Its latest report provides a wealth of data on U.S. prescription drug use.
Continuing a major trend, IMS finds that 78% of the nearly 4 billion U.S. prescriptions written in 2010 were for generic drugs (both unbranded and those still sold under a brand name). In order of number of prescriptions written in 2010, the 10 most-prescribed drugs in the U.S. are:
•Hydrocodone (combined with acetaminophen) -- 131.2 million prescriptions
•Generic Zocor (simvastatin), a cholesterol-lowering statin drug -- 94.1 million prescriptions
•Lisinopril (brand names include Prinivil and Zestril), a blood pressure drug -- 87.4 million prescriptions
•Generic Synthroid (levothyroxine sodium), synthetic thyroid hormone -- 70.5 million prescriptions
•Generic Norvasc (amlodipine besylate), an angina/blood pressure drug -- 57.2 million prescriptions
•Generic Prilosec (omeprazole), an antiacid drug -- 53.4 million prescriptions (does not include over-the-counter sales)
•Azithromycin (brand names include Z-Pak and Zithromax), an antibiotic -- 52.6 million prescriptions
•Amoxicillin (various brand names), an antibiotic -- 52.3 million prescriptions
•Generic Glucophage (metformin), a diabetes drug -- 48.3 million prescriptions
•Hydrochlorothiazide (various brand names), a water pill used to lower blood pressure -- 47.8 million prescriptions.
The 10 Best-Selling Drugs
It shouldn't be a surprise that these generic drugs are not the ones bringing in the big bucks for pharmaceutical companies. The drugs on which we spend the most money are those that are still new enough to be protected against generic competition.
The IMS reports that Americans spent $307 billion on prescription drugs in 2010. The 10 drugs on which we spent the most were:
•Lipitor, a cholesterol-lowering statin drug -- $7.2 billion
•Nexium, an antacid drug -- $6.3 billion
•Plavix, a blood thinner -- $6.1 billion
•Advair Diskus, an asthma inhaler -- $4.7 billion
•Abilify, an antipsychotic drug -- $4.6 billion
•Seroquel, an antipsychotic drug -- $4.4 billion
•Singulair, an oral asthma drug -- $4.1 billion
•Crestor, a cholesterol-lowering statin drug -- $3.8 billion
•Actos, a diabetes drug -- $3.5 billion
•Epogen, an injectable anemia drug -- $3.3 billion
U.S. Prescription Drug Use: 2010 Factoids
Who's paying for all these drugs? Commercial insurance helped pay for 63% of prescriptions, down from 66% five years ago. Federal government spending through Medicare Part D covered 22% of prescriptions.
For Americans covered by insurance, Medicare, or Medicaid, the average co-payment for a prescription was $10.73 -- down a bit from 2009 due to increased use of generic drugs. The average co-payment for branded drugs for which generic alternatives were available jumped 6% to $22.73.
Other facts from the 2010 IMS report:
•Doctor visits were down 4.2% since 2009.
•Patients filled more than half of their prescriptions -- 54% -- at chain drugstores, possibly because of discounts on generic drugs.
•Brands that lost their protection from generic competition led to $12.6 billion less spending in 2010 than in 2009.
•The price increase for drugs without generic competition led to $16.6 billion more spending in 2010 than in 2009.
•Drug companies offered $4.5 billion in rebates to assist patients with the high cost of brand name drugs for which there was no generic alternative.
SOURCE:
IMS Institute for Healthcare Informatics: "The Use of Medicines in the United States: Review of 2010," April 2011.
April 20, 2011 — The 10 most prescribed drugs in the U.S. aren't the drugs on which we spend the most, according to a report from the IMS Institute for Healthcare Informatics.
The institute is the public face of IMS, a pharmaceutical market intelligence firm. Its latest report provides a wealth of data on U.S. prescription drug use.
Continuing a major trend, IMS finds that 78% of the nearly 4 billion U.S. prescriptions written in 2010 were for generic drugs (both unbranded and those still sold under a brand name). In order of number of prescriptions written in 2010, the 10 most-prescribed drugs in the U.S. are:
•Hydrocodone (combined with acetaminophen) -- 131.2 million prescriptions
•Generic Zocor (simvastatin), a cholesterol-lowering statin drug -- 94.1 million prescriptions
•Lisinopril (brand names include Prinivil and Zestril), a blood pressure drug -- 87.4 million prescriptions
•Generic Synthroid (levothyroxine sodium), synthetic thyroid hormone -- 70.5 million prescriptions
•Generic Norvasc (amlodipine besylate), an angina/blood pressure drug -- 57.2 million prescriptions
•Generic Prilosec (omeprazole), an antiacid drug -- 53.4 million prescriptions (does not include over-the-counter sales)
•Azithromycin (brand names include Z-Pak and Zithromax), an antibiotic -- 52.6 million prescriptions
•Amoxicillin (various brand names), an antibiotic -- 52.3 million prescriptions
•Generic Glucophage (metformin), a diabetes drug -- 48.3 million prescriptions
•Hydrochlorothiazide (various brand names), a water pill used to lower blood pressure -- 47.8 million prescriptions.
The 10 Best-Selling Drugs
It shouldn't be a surprise that these generic drugs are not the ones bringing in the big bucks for pharmaceutical companies. The drugs on which we spend the most money are those that are still new enough to be protected against generic competition.
The IMS reports that Americans spent $307 billion on prescription drugs in 2010. The 10 drugs on which we spent the most were:
•Lipitor, a cholesterol-lowering statin drug -- $7.2 billion
•Nexium, an antacid drug -- $6.3 billion
•Plavix, a blood thinner -- $6.1 billion
•Advair Diskus, an asthma inhaler -- $4.7 billion
•Abilify, an antipsychotic drug -- $4.6 billion
•Seroquel, an antipsychotic drug -- $4.4 billion
•Singulair, an oral asthma drug -- $4.1 billion
•Crestor, a cholesterol-lowering statin drug -- $3.8 billion
•Actos, a diabetes drug -- $3.5 billion
•Epogen, an injectable anemia drug -- $3.3 billion
U.S. Prescription Drug Use: 2010 Factoids
Who's paying for all these drugs? Commercial insurance helped pay for 63% of prescriptions, down from 66% five years ago. Federal government spending through Medicare Part D covered 22% of prescriptions.
For Americans covered by insurance, Medicare, or Medicaid, the average co-payment for a prescription was $10.73 -- down a bit from 2009 due to increased use of generic drugs. The average co-payment for branded drugs for which generic alternatives were available jumped 6% to $22.73.
Other facts from the 2010 IMS report:
•Doctor visits were down 4.2% since 2009.
•Patients filled more than half of their prescriptions -- 54% -- at chain drugstores, possibly because of discounts on generic drugs.
•Brands that lost their protection from generic competition led to $12.6 billion less spending in 2010 than in 2009.
•The price increase for drugs without generic competition led to $16.6 billion more spending in 2010 than in 2009.
•Drug companies offered $4.5 billion in rebates to assist patients with the high cost of brand name drugs for which there was no generic alternative.
SOURCE:
IMS Institute for Healthcare Informatics: "The Use of Medicines in the United States: Review of 2010," April 2011.
viernes, 6 de mayo de 2011
NDM-1: A Local Clone Emerges with Worldwide Aspirations
Andrea Marra
Abstract
Are bacteria always going to outsmart us? With the emergence of the metallo-β-lactamase blaNDM-1 gene, it certainly seems so. Whereas at one time bacterial clones resided in hospitals or long-term care facilities, it is now apparent that they have the capability of thriving in the community and quickly spreading across countries and continents with few impediments, thanks to accessible, rapid global travel. Thus, under conditions favoring the organism (promiscuous or inappropriate antibiotic use and poor infection control procedures), what was at one time a local problem can rapidly become a worldwide health crisis. Given that the discovery and development of a new antibiotic can take a decade or more, multiply resistant pathogens can have ample time to wreak havoc before a successful novel agent comes to market. At one time a single drug, penicillin, was enough to raise expectations that new antibiotics were unnecessary; we have since seen that bacteria can generate stable resistance to every antibiotic in rapid fashion, with no detrimental effects on their pathogenicity.
Introduction
The antibiotic resistance determinant bla NDM-1 is a prime example of the ability of bacteria to accumulate resistance determinants, maintain infectivity and fitness and spread rapidly around the world. NDM-1 (New Delhi Metallo-β-lactamase-1) is a novel plasmid-borne metallo-β-lactamase (MBL) that has so far been isolated only in Enterobacteriaceae.[1] The identification of this resistance determinant was first reported in 2008;[2] by 2010,[1] it had spread from its base in New Delhi, India across the Indian subcontinent to Pakistan and Bangladesh and also in Australia and throughout the US, the UK, France and Canada.[1–7]
The bla NDM-1 gene was isolated from Klebsiella pneumoniae and Escherichia coli cultures from the same patient suffering from a urinary tract infection (UTI); the organisms were found to be resistant to all antibiotic classes with the exception of colistin.[2] Whereas NDM-1 shares the ability to hydrolyze carbapenems with other MBLs, several features make it especially worrisome. First, that this gene was found in K. pneumoniae and E. coli isolated from the same patient raises the possibility of facile horizontal resistance gene transfer in vivo; second, that it was found in Enterobacteriaceae suggests the opportunity for broad community dissemination; third, elevated MICs to many antibiotic classes indicates potential for widespread treatment failure as these organisms spread; and finally, the gene's location on mobile genetic elements allows rapid spread between pathogens with no effective antibiotics to counteract it.[1–3,8]
The generations of β-lactam antibiotics came about due to the steady evolution of bacterial resistance mechanisms that disable them. β-lactam agents (cephalosporins, carbapenems, clavulanate-type β-lactamase inhibitors, monobactams and penems) bind penicillin-binding proteins, which are involved in bacterial cell wall peptidoglycan biosynthesis; resistance to them is achieved by their efflux out of the cell, alteration of the penicillin-binding protein target, decreased porin production to limit cell entry or hydrolysis of the β-lactam itself.[9–14] One count estimates the number of different β-lactamases produced by disparate organisms to be over 950,[11] with four major classes: penicillinases, cephalosporinases (known as AmpC-type), extended-spectrum β-lactamases (ESBLs) and carbapenemases, with the latter two moving to the forefront in recent years (Table 1).[10,11,15–17] Until approximately a decade ago, ESBL-producing isolates were confined to hospitals, with K. pneumoniae being the main culprit. At present, however, community-acquired infections are of major concern, particularly UTIs, caused by ESBL-positive K. pneumoniae as well as E. coli, with different so-called CTX-M ESBLs predominating in different countries.[17,18] The number, diversity and rate of occurrence of ESBLs targeting penicillins and cephalosporins has led to more frequent use of carbapenems, which in turn has resulted in increased carbapenem resistance due to both serine carbapenemases and MBLs. For example, in India, 70–90% of Enterobacteriaceae are ESBL-producers; as a result, carbapenem use to treat these infections has increased, and so has carbapenem resistance.[1,18] In addition, in the UK at least, greater than 80% of ESBL-producing E. coli from bloodstream infections are fluoroquinolone-resistant, and greater than 40% are gentamicin-resistant, thus presenting more challenges to clinicians.[8] The serine carbapenemases found in Enterobacteriaceae, particularly in K. pneumoniae, are now endemic worldwide, with the plasmid-borne K. pneumoniae carbapenemases being the most prevalent.[11] The rapidly emerging MBLs are of concern because of their broad β-lactam resistance coverage and their transmissibility: most of the MBLs within clinical Enterobacteriaceae isolates are contained within gene cassettes on integrons.[11,15] The integron itself encodes an integrase gene, a recombination site and a promoter for expression of the gene contained therein, and often resistance elements for antiseptics (qac) and sulfonamide (sul).[15] Structures such as integrons allow facile transmission within the same cell, either into the chromosome or onto plasmids – once these elements are located on plasmids, transfer between cells becomes a matter of opportunity and pressure as antibiotic use encourages selection of resistance.
The bla NDM-1 gene is contained within such structures and perhaps not surprisingly has spread around the world rapidly from its likely origins on the Indian subcontinent. Although Yong et al. [2] have demonstrated that NDM-1 is not as robust as other MBLs (such as VIM-2 and IMP-1) in terms of binding to and hydrolysis of cephalosporins and penicillins, its real threat is its plasmid-borne location and the ease with which it is able to spread.[1–3,7] The K. pneumoniae isolate bearing bla NDM-1 carries three antibiotic resistance regions:[2] one region encodes NDM-1, plus an efflux pump and the bla DHA AmpC gene; the second encodes a rifampicin resistance gene (arr2), a novel erythromycin esterase gene (ereC), the gene CM1A7 (encoding chloramphenicol resistance), and qac/sul; and the third region contains yet another AmpC gene, bla CMY4.[2]
Sequencing of the original patient isolate revealed that the bla NDM-1 gene encoded a 27.5 kDa protein and had a lower G+C content than surrounding DNA, suggesting a non-Klebsiella origin. Kumarasamy et al. [1] molecularly characterized and studied the epidemiology of the NDM-1 isolates. For their analysis, the authors examined carbapenem-resistant Enterobacteriaceae from different sites on the Indian subcontinent and in the UK, analyzing the isolates by MIC, pulsed-field gel electrophoresis and PCR. The clinical isolates from India were primarily from two cities, Chennai and Haryana, in addition to isolates collected from three other cities in India and eight cities in Pakistan; the movement of this gene via patient travel was also investigated as isolates in the UK were screened for its presence. The majority of infections from which the NDM-1 Enterobacteriaceae were isolated were community-acquired UTIs, pneumonia and bloodstream infections.
The antibiograms of the NDM-1-producing isolates from the disparate sites told a disturbing story. In addition to being carbapenem-resistant, all the isolates were found to be nonsusceptible to several antibiotic classes. Since only a small number of isolates (all from the UK) remained susceptible to aztreonam (which is not hydrolyzed by MBLs including NDM-1), it became clear that the rest contained genes encoding ESBLs and/or the β-lactamase AmpC; indeed, sequence analysis revealed that the majority of the isolates carried bla CTX-M-15 and bla CMY-4. The UK isolates were also resistant to tobramycin, minocycline and amikacin, and most were resistant to ciprofloxacin and gentamicin as well. A total of 89% of the UK isolates were susceptible to colistin, and 64% were susceptible to tigecycline.[1]
The isolates from Chennai and Haryana followed a similar pattern: nearly all were resistant to all β-lactams (including aztreonam), fluoroquinolones, aminoglycosides and minocycline, with greater than 50% susceptible to tigecycline and all susceptible to colistin. Interestingly, all of the Haryana isolates were clonal, perhaps suggesting an increased fitness of this particular isolate. Most worrisome is the single K. pneumoniae isolate from Chennai that manifested resistance to all of the antibiotic classes mentioned above, with a MIC of colistin of 32 µg/ml and of tigecycline of 8 µg/ml. Though panresistant isolates such as this are rare, they have been reported in Greece[19,20] and significant spread would mean a return to the preantibiotic era.
As noted above, the presence of bla NDM-1 does not in itself result in the broad β-lactam resistance that is observed in all of these isolates; the major concern with this resistance determinant is the piggy-backing of this gene onto plasmids or transposons or integrons, mobile genetic elements that allow easy transfer between organisms (in the case of plasmids) or between plasmids and chromosomes (transposons and integrons). Indeed, when all of the Chennai, Haryana and UK isolates were analyzed for the presence of plasmids and the location of bla NDM-1, they were all found to harbor multiple plasmids (50–500 kb in size, in some cases as many as eight) and the gene was plasmid-borne in all of them. In addition, the bla NDM-1 gene was also in the chromosomes of three of the UK isolates and on more than one plasmid in others, reinforcing the idea that this gene is capable of movement within the same cell.[1]
The analysis by Kumarasamy et al. included transfer of plasmids from isolates to an E. coli laboratory strain.[1] The results suggest high transmissibility of bla NDM-1-carrying plasmids between Enterobacteriaceae; surely selective pressure through inappropriate or nonprescription use of antibiotics (and especially carbapenems) in India[1,21] as well as Greece[22] could select for this resistance element and significantly increase the numbers of bla NDM-1 isolates from patients. As this antibiotic resistance element is not confined to hospitals or long-term care facilities, but was for the most part isolated from community-acquired infections, infection control measures would be extremely difficult to implement.
Conclusion & Future Perspective
The carbapenems were developed to combat the increasing prevalence of β-lactamase enzymes in Gram-negative organisms and have become first-line therapy against serious infections. The wide-ranging and rapid emergence of a resistance element such as bla NDM-1 is cause for concern and has justifiably elicited coverage in the scientific and medical communities. The MBLs present many challenges to clinicians as well as drug developers: they have an incredibly broad range of activity against β-lactams, they are resistant to β-lactamase inhibitors, and they are commonly linked to aminoglycoside resistance genes. The isolates studied so far appear to remain susceptible to tigecycline and colistin, but both drugs have clinical limitations – tigecycline achieves low serum and urine levels and thus is not routinely prescribed for bacteremia or UTI; colistin has good efficacy in general but is weak against respiratory tract infections.[8,9,15,23] Ironically, the NDM-1 isolates were predominantly from patients suffering from UTIs, respiratory tract infections and bloodstream infections. Since its emergence in the UK in 2008, carbapenem-resistant Enterobacteriaceae carrying the bla NDM-1 gene have become the predominant Enterobacteriaceae carbapenem resistance determinant – from approximately 5% in 2008 to 44% in 2009.[1] In that time period, 37 bla NDM-1- producing Enterobacteriaceae were isolated from 25 sites across England, Scotland and Northern Ireland. More than half of the 29 patients involved had traveled to India or Pakistan within the previous year, and nearly half had been admitted to a hospital while there. The isolates were determined to be K. pneumoniae, E. coli, Enterobacter spp., Citrobacter freundii, Morganella morganii, and Providencia spp..[23] The isolation of this gene in Enterobacteriaceae – common flora – on mobile genetic elements, from patients in varied clinical settings around the world should herald a worldwide call for the development of global guidelines involving testing, treatment and infection control measures to contain what could become a serious global health issue.
References
1.Kumarasamy KK, Toleman MA, Walsh TR et al.: Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study. Lancet Infect. Dis. 10(9), 597–602 (2010).
2.Yong D, Toleman MA, Giske CG et al.: Characterization of a new metallo-β-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob. Agents Chemother. 3(12), 5046–5054 (2009).
3.Deshpande P, Rodrigues C, Shetty A et al.: New Delhi metallo-β-lactamase (NDM-1) in Enterobacteriaceae: treatment options with carbapenems compromised. J. Assoc. Phys. India 58, 147–149 (2010).
4.Poirel L, Lagrutta E, Taylor P, Pham J, Nordmann P: Emergence of metallo-β-lactamase NDM-1-producing multidrug-resistant Escherichia coli in Australia. Antimicrob. Agents Chemother. 54(11), 4914–4916 (2010).
5.Centers for Disease Control and Prevention: Detection of Enterobacteriaceae isolates carrying metallo-β-lactamase–United States, 2010. MMWR Morb. Mortal Wkly Rep. 59, 750 (2010).
6.Poirel L, Ros A, Carricajo A et al.: Extremely drug-resistant Citrobacter freundii identified in a patient returning from India and producing NDM-1 and other carbapenemases. Antimicrob. Agents Chemother. 55(1), 447–448 (2011).
7.Webster PC: Global action urged in response to new breed of drug-resistant bacteria. Can. Med. Assoc. J. 182(15), 1602–1603 (2010).
8.Livermore D: Has the era of untreatable infections arrived? J. Antimicrob. Chemother. 64(Suppl. 1),I29–I36 (2009).
9.Llarrull LI, Testero SA, Fisher JF, Mobashery S: The future of the β-lactams. Curr. Opin. Microbiol. 13, 551–557 (2010).
10.Thomson KS: Extended-spectrum-β-lactamase, AmpC, and carbapenemase issues. J. Clin. Microbiol. 48(4), 1019–1025 (2010).
11.Bush K: Alarming β-lactamase-mediated resistance in multidrug-resistant Enterobacteriaceae. Curr. Opin. Microbiol. 13, 558–564 (2010).
12.Garcia-Fernandez A, Miriagou V, Papagiannitis CC et al.: An ertapenem-resistant extended-spectrum-β-lactamase-producing Klebsiella pneumonia clone carries a novel OmpK36 porin variant. Antimicrob. Agents Chemother. 54(10), 4178–4184 (2010).
13.Kitchel B, Rasheed JK, Endiami A et al.: Genetic factors associated with elevated carbapenem resistance in KPC-producing Klebsiella pneumoniae. Antimicrob. Agents Chemother. 54(10), 4201–4207 (2010).
14.Mammeri H, Guillon H, Eb F, Nordmann P: Phenotypic and biochemical comparison of the carbapenem-hydrolyzing activities of five plasmid-borne AmpC β-lactamases. Antimicrob. Agents Chemother. 54(11), 4556–4560 (2010).
15.Walsh TR, Toleman MA, Poirel L, Nordmann P: Metallo-β-lactamases: the quiet before the storm? Clin. Microbiol. Rev. 18(2), 306–325 (2005).
16.Nordmann P, Cuzon G, Naas T: The real threat of Klebsiella pneumoniae carbapenemase-producing bacteria. Lancet 9, 228–236 (2009).
17.Hawkey PM, Jones AM: The changing epidemiology of resistance. J. Antimicrob. Chemother. 64(Suppl. 1),I3–I10 (2009).
18.Hawkey PM: Prevalence and clonality of extended-spectrum β-lactamases in Asia. Clin. Microbiol. Infect. 14(Suppl. 1), 159–165 (2008).
19.Souli M, Kontopidou FV, Koratzanis E et al.: In vitro activity of tigecycline against multiple-drug-resistant, including pan-resistant, Gram-negative and Gram-positive clinical isolates from Greek hospitals. Antimicrob. Agents Chemother. 50(9), 3166–3169 (2006).
20.Antoniadou A, Kontopidou F, Poulakou G et al.: Colistin-resistant isolates of Klebsiella pneumoniae emerging in intensive care unit patients: first report of a multiclonal cluster. J. Antimicrob. Chemother. 59, 786–790 (2007).
21.Krishna B: New Delhi metallo-β-lactamases: a wake-up call for microbiologists. Indian J. Med. Microbiol. 28, 265–266 (2010).
22.Plachouras D, Kavatha D, Antoniadou A et al.: Dispensing of antibiotics without prescription in Greece, 2008: another link in the antibiotic resistance chain. Euro Surveill. 15(7), 19488 (2010).
23.Health Protection Agency: Current news: multi-resistant hospital bacteria linked to India and Pakistan. Health Protection Report 3(26), 3–4 (2009).
24.Bertini A, Poirel L, Bernabeu S et al.: Multicopy blaOXA-58 gene as a source of high-level resistance to carbapenems in Acinetobacter baumannii. Antimicrob. Agents Chemother. 51(7), 2324–2328 (2007).
Andrea Marra
Rib-X Pharmaceuticals, Inc., 300 George Street, Suite 301, New Haven, CT 06511, USA. Tel.: +1 203 848 3349 Fax: +1 203 624 5627 amarra@rib-x.com
Acknowledgements
The author would like to acknowledge the expert technical advice and guidance of Tom Gootz in the preparation of this manuscript.
Financial & competing interests disclosure
The author has no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.
No writing assistance was utilized in the production of this manuscript.
Future Microbiology. 2011;6(2):137-141. © 2011 Future Medicine Ltd
Abstract
Are bacteria always going to outsmart us? With the emergence of the metallo-β-lactamase blaNDM-1 gene, it certainly seems so. Whereas at one time bacterial clones resided in hospitals or long-term care facilities, it is now apparent that they have the capability of thriving in the community and quickly spreading across countries and continents with few impediments, thanks to accessible, rapid global travel. Thus, under conditions favoring the organism (promiscuous or inappropriate antibiotic use and poor infection control procedures), what was at one time a local problem can rapidly become a worldwide health crisis. Given that the discovery and development of a new antibiotic can take a decade or more, multiply resistant pathogens can have ample time to wreak havoc before a successful novel agent comes to market. At one time a single drug, penicillin, was enough to raise expectations that new antibiotics were unnecessary; we have since seen that bacteria can generate stable resistance to every antibiotic in rapid fashion, with no detrimental effects on their pathogenicity.
Introduction
The antibiotic resistance determinant bla NDM-1 is a prime example of the ability of bacteria to accumulate resistance determinants, maintain infectivity and fitness and spread rapidly around the world. NDM-1 (New Delhi Metallo-β-lactamase-1) is a novel plasmid-borne metallo-β-lactamase (MBL) that has so far been isolated only in Enterobacteriaceae.[1] The identification of this resistance determinant was first reported in 2008;[2] by 2010,[1] it had spread from its base in New Delhi, India across the Indian subcontinent to Pakistan and Bangladesh and also in Australia and throughout the US, the UK, France and Canada.[1–7]
The bla NDM-1 gene was isolated from Klebsiella pneumoniae and Escherichia coli cultures from the same patient suffering from a urinary tract infection (UTI); the organisms were found to be resistant to all antibiotic classes with the exception of colistin.[2] Whereas NDM-1 shares the ability to hydrolyze carbapenems with other MBLs, several features make it especially worrisome. First, that this gene was found in K. pneumoniae and E. coli isolated from the same patient raises the possibility of facile horizontal resistance gene transfer in vivo; second, that it was found in Enterobacteriaceae suggests the opportunity for broad community dissemination; third, elevated MICs to many antibiotic classes indicates potential for widespread treatment failure as these organisms spread; and finally, the gene's location on mobile genetic elements allows rapid spread between pathogens with no effective antibiotics to counteract it.[1–3,8]
The generations of β-lactam antibiotics came about due to the steady evolution of bacterial resistance mechanisms that disable them. β-lactam agents (cephalosporins, carbapenems, clavulanate-type β-lactamase inhibitors, monobactams and penems) bind penicillin-binding proteins, which are involved in bacterial cell wall peptidoglycan biosynthesis; resistance to them is achieved by their efflux out of the cell, alteration of the penicillin-binding protein target, decreased porin production to limit cell entry or hydrolysis of the β-lactam itself.[9–14] One count estimates the number of different β-lactamases produced by disparate organisms to be over 950,[11] with four major classes: penicillinases, cephalosporinases (known as AmpC-type), extended-spectrum β-lactamases (ESBLs) and carbapenemases, with the latter two moving to the forefront in recent years (Table 1).[10,11,15–17] Until approximately a decade ago, ESBL-producing isolates were confined to hospitals, with K. pneumoniae being the main culprit. At present, however, community-acquired infections are of major concern, particularly UTIs, caused by ESBL-positive K. pneumoniae as well as E. coli, with different so-called CTX-M ESBLs predominating in different countries.[17,18] The number, diversity and rate of occurrence of ESBLs targeting penicillins and cephalosporins has led to more frequent use of carbapenems, which in turn has resulted in increased carbapenem resistance due to both serine carbapenemases and MBLs. For example, in India, 70–90% of Enterobacteriaceae are ESBL-producers; as a result, carbapenem use to treat these infections has increased, and so has carbapenem resistance.[1,18] In addition, in the UK at least, greater than 80% of ESBL-producing E. coli from bloodstream infections are fluoroquinolone-resistant, and greater than 40% are gentamicin-resistant, thus presenting more challenges to clinicians.[8] The serine carbapenemases found in Enterobacteriaceae, particularly in K. pneumoniae, are now endemic worldwide, with the plasmid-borne K. pneumoniae carbapenemases being the most prevalent.[11] The rapidly emerging MBLs are of concern because of their broad β-lactam resistance coverage and their transmissibility: most of the MBLs within clinical Enterobacteriaceae isolates are contained within gene cassettes on integrons.[11,15] The integron itself encodes an integrase gene, a recombination site and a promoter for expression of the gene contained therein, and often resistance elements for antiseptics (qac) and sulfonamide (sul).[15] Structures such as integrons allow facile transmission within the same cell, either into the chromosome or onto plasmids – once these elements are located on plasmids, transfer between cells becomes a matter of opportunity and pressure as antibiotic use encourages selection of resistance.
The bla NDM-1 gene is contained within such structures and perhaps not surprisingly has spread around the world rapidly from its likely origins on the Indian subcontinent. Although Yong et al. [2] have demonstrated that NDM-1 is not as robust as other MBLs (such as VIM-2 and IMP-1) in terms of binding to and hydrolysis of cephalosporins and penicillins, its real threat is its plasmid-borne location and the ease with which it is able to spread.[1–3,7] The K. pneumoniae isolate bearing bla NDM-1 carries three antibiotic resistance regions:[2] one region encodes NDM-1, plus an efflux pump and the bla DHA AmpC gene; the second encodes a rifampicin resistance gene (arr2), a novel erythromycin esterase gene (ereC), the gene CM1A7 (encoding chloramphenicol resistance), and qac/sul; and the third region contains yet another AmpC gene, bla CMY4.[2]
Sequencing of the original patient isolate revealed that the bla NDM-1 gene encoded a 27.5 kDa protein and had a lower G+C content than surrounding DNA, suggesting a non-Klebsiella origin. Kumarasamy et al. [1] molecularly characterized and studied the epidemiology of the NDM-1 isolates. For their analysis, the authors examined carbapenem-resistant Enterobacteriaceae from different sites on the Indian subcontinent and in the UK, analyzing the isolates by MIC, pulsed-field gel electrophoresis and PCR. The clinical isolates from India were primarily from two cities, Chennai and Haryana, in addition to isolates collected from three other cities in India and eight cities in Pakistan; the movement of this gene via patient travel was also investigated as isolates in the UK were screened for its presence. The majority of infections from which the NDM-1 Enterobacteriaceae were isolated were community-acquired UTIs, pneumonia and bloodstream infections.
The antibiograms of the NDM-1-producing isolates from the disparate sites told a disturbing story. In addition to being carbapenem-resistant, all the isolates were found to be nonsusceptible to several antibiotic classes. Since only a small number of isolates (all from the UK) remained susceptible to aztreonam (which is not hydrolyzed by MBLs including NDM-1), it became clear that the rest contained genes encoding ESBLs and/or the β-lactamase AmpC; indeed, sequence analysis revealed that the majority of the isolates carried bla CTX-M-15 and bla CMY-4. The UK isolates were also resistant to tobramycin, minocycline and amikacin, and most were resistant to ciprofloxacin and gentamicin as well. A total of 89% of the UK isolates were susceptible to colistin, and 64% were susceptible to tigecycline.[1]
The isolates from Chennai and Haryana followed a similar pattern: nearly all were resistant to all β-lactams (including aztreonam), fluoroquinolones, aminoglycosides and minocycline, with greater than 50% susceptible to tigecycline and all susceptible to colistin. Interestingly, all of the Haryana isolates were clonal, perhaps suggesting an increased fitness of this particular isolate. Most worrisome is the single K. pneumoniae isolate from Chennai that manifested resistance to all of the antibiotic classes mentioned above, with a MIC of colistin of 32 µg/ml and of tigecycline of 8 µg/ml. Though panresistant isolates such as this are rare, they have been reported in Greece[19,20] and significant spread would mean a return to the preantibiotic era.
As noted above, the presence of bla NDM-1 does not in itself result in the broad β-lactam resistance that is observed in all of these isolates; the major concern with this resistance determinant is the piggy-backing of this gene onto plasmids or transposons or integrons, mobile genetic elements that allow easy transfer between organisms (in the case of plasmids) or between plasmids and chromosomes (transposons and integrons). Indeed, when all of the Chennai, Haryana and UK isolates were analyzed for the presence of plasmids and the location of bla NDM-1, they were all found to harbor multiple plasmids (50–500 kb in size, in some cases as many as eight) and the gene was plasmid-borne in all of them. In addition, the bla NDM-1 gene was also in the chromosomes of three of the UK isolates and on more than one plasmid in others, reinforcing the idea that this gene is capable of movement within the same cell.[1]
The analysis by Kumarasamy et al. included transfer of plasmids from isolates to an E. coli laboratory strain.[1] The results suggest high transmissibility of bla NDM-1-carrying plasmids between Enterobacteriaceae; surely selective pressure through inappropriate or nonprescription use of antibiotics (and especially carbapenems) in India[1,21] as well as Greece[22] could select for this resistance element and significantly increase the numbers of bla NDM-1 isolates from patients. As this antibiotic resistance element is not confined to hospitals or long-term care facilities, but was for the most part isolated from community-acquired infections, infection control measures would be extremely difficult to implement.
Conclusion & Future Perspective
The carbapenems were developed to combat the increasing prevalence of β-lactamase enzymes in Gram-negative organisms and have become first-line therapy against serious infections. The wide-ranging and rapid emergence of a resistance element such as bla NDM-1 is cause for concern and has justifiably elicited coverage in the scientific and medical communities. The MBLs present many challenges to clinicians as well as drug developers: they have an incredibly broad range of activity against β-lactams, they are resistant to β-lactamase inhibitors, and they are commonly linked to aminoglycoside resistance genes. The isolates studied so far appear to remain susceptible to tigecycline and colistin, but both drugs have clinical limitations – tigecycline achieves low serum and urine levels and thus is not routinely prescribed for bacteremia or UTI; colistin has good efficacy in general but is weak against respiratory tract infections.[8,9,15,23] Ironically, the NDM-1 isolates were predominantly from patients suffering from UTIs, respiratory tract infections and bloodstream infections. Since its emergence in the UK in 2008, carbapenem-resistant Enterobacteriaceae carrying the bla NDM-1 gene have become the predominant Enterobacteriaceae carbapenem resistance determinant – from approximately 5% in 2008 to 44% in 2009.[1] In that time period, 37 bla NDM-1- producing Enterobacteriaceae were isolated from 25 sites across England, Scotland and Northern Ireland. More than half of the 29 patients involved had traveled to India or Pakistan within the previous year, and nearly half had been admitted to a hospital while there. The isolates were determined to be K. pneumoniae, E. coli, Enterobacter spp., Citrobacter freundii, Morganella morganii, and Providencia spp..[23] The isolation of this gene in Enterobacteriaceae – common flora – on mobile genetic elements, from patients in varied clinical settings around the world should herald a worldwide call for the development of global guidelines involving testing, treatment and infection control measures to contain what could become a serious global health issue.
References
1.Kumarasamy KK, Toleman MA, Walsh TR et al.: Emergence of a new antibiotic resistance mechanism in India, Pakistan, and the UK: a molecular, biological, and epidemiological study. Lancet Infect. Dis. 10(9), 597–602 (2010).
2.Yong D, Toleman MA, Giske CG et al.: Characterization of a new metallo-β-lactamase gene, blaNDM-1, and a novel erythromycin esterase gene carried on a unique genetic structure in Klebsiella pneumoniae sequence type 14 from India. Antimicrob. Agents Chemother. 3(12), 5046–5054 (2009).
3.Deshpande P, Rodrigues C, Shetty A et al.: New Delhi metallo-β-lactamase (NDM-1) in Enterobacteriaceae: treatment options with carbapenems compromised. J. Assoc. Phys. India 58, 147–149 (2010).
4.Poirel L, Lagrutta E, Taylor P, Pham J, Nordmann P: Emergence of metallo-β-lactamase NDM-1-producing multidrug-resistant Escherichia coli in Australia. Antimicrob. Agents Chemother. 54(11), 4914–4916 (2010).
5.Centers for Disease Control and Prevention: Detection of Enterobacteriaceae isolates carrying metallo-β-lactamase–United States, 2010. MMWR Morb. Mortal Wkly Rep. 59, 750 (2010).
6.Poirel L, Ros A, Carricajo A et al.: Extremely drug-resistant Citrobacter freundii identified in a patient returning from India and producing NDM-1 and other carbapenemases. Antimicrob. Agents Chemother. 55(1), 447–448 (2011).
7.Webster PC: Global action urged in response to new breed of drug-resistant bacteria. Can. Med. Assoc. J. 182(15), 1602–1603 (2010).
8.Livermore D: Has the era of untreatable infections arrived? J. Antimicrob. Chemother. 64(Suppl. 1),I29–I36 (2009).
9.Llarrull LI, Testero SA, Fisher JF, Mobashery S: The future of the β-lactams. Curr. Opin. Microbiol. 13, 551–557 (2010).
10.Thomson KS: Extended-spectrum-β-lactamase, AmpC, and carbapenemase issues. J. Clin. Microbiol. 48(4), 1019–1025 (2010).
11.Bush K: Alarming β-lactamase-mediated resistance in multidrug-resistant Enterobacteriaceae. Curr. Opin. Microbiol. 13, 558–564 (2010).
12.Garcia-Fernandez A, Miriagou V, Papagiannitis CC et al.: An ertapenem-resistant extended-spectrum-β-lactamase-producing Klebsiella pneumonia clone carries a novel OmpK36 porin variant. Antimicrob. Agents Chemother. 54(10), 4178–4184 (2010).
13.Kitchel B, Rasheed JK, Endiami A et al.: Genetic factors associated with elevated carbapenem resistance in KPC-producing Klebsiella pneumoniae. Antimicrob. Agents Chemother. 54(10), 4201–4207 (2010).
14.Mammeri H, Guillon H, Eb F, Nordmann P: Phenotypic and biochemical comparison of the carbapenem-hydrolyzing activities of five plasmid-borne AmpC β-lactamases. Antimicrob. Agents Chemother. 54(11), 4556–4560 (2010).
15.Walsh TR, Toleman MA, Poirel L, Nordmann P: Metallo-β-lactamases: the quiet before the storm? Clin. Microbiol. Rev. 18(2), 306–325 (2005).
16.Nordmann P, Cuzon G, Naas T: The real threat of Klebsiella pneumoniae carbapenemase-producing bacteria. Lancet 9, 228–236 (2009).
17.Hawkey PM, Jones AM: The changing epidemiology of resistance. J. Antimicrob. Chemother. 64(Suppl. 1),I3–I10 (2009).
18.Hawkey PM: Prevalence and clonality of extended-spectrum β-lactamases in Asia. Clin. Microbiol. Infect. 14(Suppl. 1), 159–165 (2008).
19.Souli M, Kontopidou FV, Koratzanis E et al.: In vitro activity of tigecycline against multiple-drug-resistant, including pan-resistant, Gram-negative and Gram-positive clinical isolates from Greek hospitals. Antimicrob. Agents Chemother. 50(9), 3166–3169 (2006).
20.Antoniadou A, Kontopidou F, Poulakou G et al.: Colistin-resistant isolates of Klebsiella pneumoniae emerging in intensive care unit patients: first report of a multiclonal cluster. J. Antimicrob. Chemother. 59, 786–790 (2007).
21.Krishna B: New Delhi metallo-β-lactamases: a wake-up call for microbiologists. Indian J. Med. Microbiol. 28, 265–266 (2010).
22.Plachouras D, Kavatha D, Antoniadou A et al.: Dispensing of antibiotics without prescription in Greece, 2008: another link in the antibiotic resistance chain. Euro Surveill. 15(7), 19488 (2010).
23.Health Protection Agency: Current news: multi-resistant hospital bacteria linked to India and Pakistan. Health Protection Report 3(26), 3–4 (2009).
24.Bertini A, Poirel L, Bernabeu S et al.: Multicopy blaOXA-58 gene as a source of high-level resistance to carbapenems in Acinetobacter baumannii. Antimicrob. Agents Chemother. 51(7), 2324–2328 (2007).
Andrea Marra
Rib-X Pharmaceuticals, Inc., 300 George Street, Suite 301, New Haven, CT 06511, USA. Tel.: +1 203 848 3349 Fax: +1 203 624 5627 amarra@rib-x.com
Acknowledgements
The author would like to acknowledge the expert technical advice and guidance of Tom Gootz in the preparation of this manuscript.
Financial & competing interests disclosure
The author has no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.
No writing assistance was utilized in the production of this manuscript.
Future Microbiology. 2011;6(2):137-141. © 2011 Future Medicine Ltd
REGLAS IMPRESCINDIBLES PARA UTILIZAR ANTIMICROBIANOS DE MANERA RACIONAL EN UNIDADES DE TERAPIA INTENSIVA
•Iniciar el tratamiento antimicrobiano precoz y de espectro adecuado a la epidemiología local en pacientes con infecciones graves, previa toma de cultivos
•Utilizar dosis y vías adecuadas a cada condición clínica y a las diferentes comorbilidades
•Conocer la distribución de patógenos prevalentes en la UTI y sus patrones de sensibilidad actualizados.
•Drenar/remover adecuadamente las colecciones o focos supurados/infecciosos (ej, catéteres vasculares)
•No tratar a los pacientes que solo estén colonizados con patógenos resistentes; proceder a su aislamiento adecuado.
•Minimizar la presión antimicrobiana promotora de resistencia bacteriana
•Utilizar con preferencia antimicrobianos con bajo potencial de generar resistencia
•No mantener tratamientos que hayan sido indicados sin criterios adecuados, como por ejemplo, persistencia de leucocitosis, infiltrados pulmonares o fiebre de bajo grado.
•Considerar desintensificar el tratamiento antimicrobiano acorde a la situación clínica y a la documentación microbiológica, reduciendo el espectro según sensibilidad.
•Establecer normas de trabajo en conjunto con los Servicios de Infectología, que incluyan la adaptación del presente consenso a la realidad local, con la participación de los efectores.
*Adaptado del Consenso SADI-SATI-INE-ADECI para el manejo racional de la antibioticoterapia en la Unidad de Terapia Intensiva.
•Utilizar dosis y vías adecuadas a cada condición clínica y a las diferentes comorbilidades
•Conocer la distribución de patógenos prevalentes en la UTI y sus patrones de sensibilidad actualizados.
•Drenar/remover adecuadamente las colecciones o focos supurados/infecciosos (ej, catéteres vasculares)
•No tratar a los pacientes que solo estén colonizados con patógenos resistentes; proceder a su aislamiento adecuado.
•Minimizar la presión antimicrobiana promotora de resistencia bacteriana
•Utilizar con preferencia antimicrobianos con bajo potencial de generar resistencia
•No mantener tratamientos que hayan sido indicados sin criterios adecuados, como por ejemplo, persistencia de leucocitosis, infiltrados pulmonares o fiebre de bajo grado.
•Considerar desintensificar el tratamiento antimicrobiano acorde a la situación clínica y a la documentación microbiológica, reduciendo el espectro según sensibilidad.
•Establecer normas de trabajo en conjunto con los Servicios de Infectología, que incluyan la adaptación del presente consenso a la realidad local, con la participación de los efectores.
*Adaptado del Consenso SADI-SATI-INE-ADECI para el manejo racional de la antibioticoterapia en la Unidad de Terapia Intensiva.
El ‘blindaje’ de las bacterias a los antibióticos, al descubierto
29 de abril de 2011 – Fuente: Science
Investigadores de la Universidad de Penn State, en Estados Unidos, han sido capaces de describir con detalle el mecanismo químico por el que una determinada cepa de bacteria ha evolucionado hasta hacerse resistente a los antibióticos, según un artículo. El profesor Squire Booker, autor del estudio, ha continuado una investigación hace unos años y, según explica, puede suponer “un paso clave” para el desarrollo de nuevos fármacos para combatir la resistencia que presentan algunas “superbacterias”, como las que a menudo se encuentran en las infecciones.
El equipo comenzó el estudio de una proteína producida por una “superbacteria”, después de que hace varios años diferentes estudios genéticos desvelasen que el Staphylococcus sciuri (un patógeno bacteriano no humano) había desarrollado un nuevo gen llamado ‘Cfr’. Según observaron, la proteína creada por este gen juega un papel clave en uno de los mecanismos de la bacteria para hacerse resistente a antibióticos.
Sucesivos estudios mostraron que el mismo gen se había cruzado con una cepa de Staphylococcus aureus, una bacteria que forma parte de la flora de las fosas nasales y la piel y causa algunas de las resistencias bacterianas más comunes.
Dado que este gen a menudo se encuentra dentro de un elemento móvil de ADN, puede pasar fácilmente de un patógeno no humano a otras especies de bacterias que sí infectan a los humanos.
“El gen, que se ha encontrado en cepas de Staphylococcus aureus en Estados Unidos, México, Brasil, España, Italia e Irlanda, hace que las bacterias se vuelvan resistentes a siete tipos de antibióticos”, explicó Booker.
Esto muestra que estas bacterias tienen “una ventaja evolutiva distinta” gracias a este gen, si bien hasta ahora no se tenía una imagen “clara” de lo que sucedía a nivel molecular.
Para resolver el misterio químico de cómo las bacterias “burlan” a tantos antibióticos, Booker y su equipo analizaron el proceso de metilación de la proteína Cfr, por el cual las enzimas añaden una pequeña etiqueta molecular en los nucleótidos (unidades estructurales de ARN y ADN).
Cuando esta etiqueta molecular se añade por una proteína llamada ‘RlmN’, se favorece la síntesis de proteínas que las bacterias necesitan para sobrevivir.
Sin embargo, ahora han observado que la proteína Cfr realiza una función idéntica a la proteína RlmN, añadiendo la etiqueta molecular en una ubicación diferente del mismo nucleótido, lo que representa “un mecanismo químico realmente nuevo en la metilación”.
Según explica Booker, el siguiente paso será utilizar esta información para diseñar nuevos compuestos que actúen junto con los antibióticos clásicos. “Ya conocemos el mecanismo específico por el cual las células bacterianas evitan algunos antibióticos, de ahí que podamos empezar a pensar cómo interrumpir el proceso, para que los antibióticos clásicos hagan su trabajo”, concluyó.
Investigadores de la Universidad de Penn State, en Estados Unidos, han sido capaces de describir con detalle el mecanismo químico por el que una determinada cepa de bacteria ha evolucionado hasta hacerse resistente a los antibióticos, según un artículo. El profesor Squire Booker, autor del estudio, ha continuado una investigación hace unos años y, según explica, puede suponer “un paso clave” para el desarrollo de nuevos fármacos para combatir la resistencia que presentan algunas “superbacterias”, como las que a menudo se encuentran en las infecciones.
El equipo comenzó el estudio de una proteína producida por una “superbacteria”, después de que hace varios años diferentes estudios genéticos desvelasen que el Staphylococcus sciuri (un patógeno bacteriano no humano) había desarrollado un nuevo gen llamado ‘Cfr’. Según observaron, la proteína creada por este gen juega un papel clave en uno de los mecanismos de la bacteria para hacerse resistente a antibióticos.
Sucesivos estudios mostraron que el mismo gen se había cruzado con una cepa de Staphylococcus aureus, una bacteria que forma parte de la flora de las fosas nasales y la piel y causa algunas de las resistencias bacterianas más comunes.
Dado que este gen a menudo se encuentra dentro de un elemento móvil de ADN, puede pasar fácilmente de un patógeno no humano a otras especies de bacterias que sí infectan a los humanos.
“El gen, que se ha encontrado en cepas de Staphylococcus aureus en Estados Unidos, México, Brasil, España, Italia e Irlanda, hace que las bacterias se vuelvan resistentes a siete tipos de antibióticos”, explicó Booker.
Esto muestra que estas bacterias tienen “una ventaja evolutiva distinta” gracias a este gen, si bien hasta ahora no se tenía una imagen “clara” de lo que sucedía a nivel molecular.
Para resolver el misterio químico de cómo las bacterias “burlan” a tantos antibióticos, Booker y su equipo analizaron el proceso de metilación de la proteína Cfr, por el cual las enzimas añaden una pequeña etiqueta molecular en los nucleótidos (unidades estructurales de ARN y ADN).
Cuando esta etiqueta molecular se añade por una proteína llamada ‘RlmN’, se favorece la síntesis de proteínas que las bacterias necesitan para sobrevivir.
Sin embargo, ahora han observado que la proteína Cfr realiza una función idéntica a la proteína RlmN, añadiendo la etiqueta molecular en una ubicación diferente del mismo nucleótido, lo que representa “un mecanismo químico realmente nuevo en la metilación”.
Según explica Booker, el siguiente paso será utilizar esta información para diseñar nuevos compuestos que actúen junto con los antibióticos clásicos. “Ya conocemos el mecanismo específico por el cual las células bacterianas evitan algunos antibióticos, de ahí que podamos empezar a pensar cómo interrumpir el proceso, para que los antibióticos clásicos hagan su trabajo”, concluyó.
martes, 15 de marzo de 2011
Resurgence of Colistin: A Review of Resistance, Toxicity, Pharmacodynamics, and Dosing
Lauren M. Lim, Pharm.D.; Neang Ly, B.S.; Dana Anderson, Pharm.D.; Jenny C. Yang, Pharm.D.; Laurie Macander, Pharm.D.; Anthony Jarkowski, III, Pharm.D.; Alan Forrest, Pharm.D.; Jurgen B. Bulitta, Ph.D.; Brian T. Tsuji, Pharm.D.
Posted: 12/09/2010; Pharmacotherapy. 2010;30(12):1279-1291. © 2010 Pharmacotherapy Publications
Abstract and Introduction
Abstract
Colistin is a polymyxin antibiotic that was discovered in the late 1940s for the treatment of gram-negative infections. After several years of clinical use, its popularity diminished because of reports of significant nephrotoxicity and neurotoxicity. Recently, the antibiotic has resurfaced as a last-line treatment option for multidrug-resistant organisms such as Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae. The need for antibiotics with coverage of these gram-negative pathogens is critical because of their high morbidity and mortality, making colistin a very important treatment option. Unfortunately, however, resistance to colistin has been documented among all three of these organisms in case reports. Although the exact mechanism causing colistin resistance has not been defined, it is hypothesized that the PmrA-PmrB and PhoP-PhoQ genetic regulatory systems may play a role. Colistin dosages must be optimized, as colistin is a last-line treatment option; in addition, suboptimal doses have been linked to the development of resistance. The lack of pharmacokinetic and pharmacodynamic studies and no universal harmonization of dose units, however, have made it difficult to derive optimal dosing regimens and specific dosing guidelines for colistin. In critically ill patients who may have multiorgan failure, renal insufficiency may alter colistin pharmacokinetics. Therefore, dosage alterations in this patient population are imperative to achieve maximal efficacy and minimal toxicity. With regard to colistin toxicity, most studies show that nephrotoxicity is reversible and less frequent than once thought, and neurotoxicity is rare. Further research is needed to fully understand the impact that the two regulatory systems have on resistance, as well as the dosages of colistin needed to inhibit and overcome these developing patterns.
Introduction
Antimicrobial resistance has become a worldwide health care crisis with many pathogens showing limited or no susceptibility to currently available antimicrobial treatments. Gram-negative infections are of even more concern because of the lack of effective treatments and the limited number of antibiotics in development to treat these potentially lethal pathogens.[1–5] No new antibiotics with activity against multidrug-resistant (MDR) gram-negative bacteria are expected to be released within the next 5 years. This emphasizes the need for last-line options, such as colistin, in cases where pathogens are resistant to all other antibiotics.
Colistin, a polymyxin antibiotic (polymyxin E), was first discovered in the 1940s but was not used clinically until the late 1950s. Historically, colistin was used to combat infections caused by problematic gram-negative bacteria. Reports of nephrotoxicity and neurotoxicity, however, deterred physicians from using the antibiotic, especially with the emergence of other antibiotics (e.g., aminoglycosides) that were less toxic. Between the 1970s and 1990s, colistin was not used often, and the number of studies analyzing its use and pharmacology was minimal.[5]
Recently, the lack of treatment options for MDR bacteria such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae, has led to the reemergence of colistin as an antimicrobial therapy. Because such a large gap exists between the years that colistin was used clinically, available pharmacokinetic and pharmacodynamic data are very limited. Thus, information regarding colistin toxicities and optimum dosing is not well defined, and no universal dosing for the antibiotic exists. In addition, reports have begun to surface of colistin resistance among the organisms that the drug is currently being used to treat.[1, 2] This increased rate of resistance has emphasized the need to provide adequate, effective dosing with minimal toxicity. To review the pharmacology, resistance, toxicities, pharmacodynamics, and dosing considerations associated with colistin, we performed a search of the MEDLINE database for journal articles published from 1945–May 2010.
Dosage Forms, Elimination, Mechanism of Action, and Spectrum of Activity
Colistin is available in two forms, colistin sulfate and colistimethate sodium, administered topically and parenterally, respectively. Both forms can be inhaled. It is extremely important to note that the two forms are not interchangeable. Colistin sulfate is cationic and stable, whereas colistimethate sodium is anionic and not stable in vitro or in vivo.[6, 7] Colistimethate sodium is the form that is safer to administer parenterally because of its lower rate of toxicity.[8] As a prodrug, colistimethate sodium is readily hydrolyzed to form partially sulfomethylated derivatives, as well as colistin sulfate, the active form of the drug.[8] This hydrolysis of colistimethate sodium to colistin is a very important step in providing the drug's antimicrobial activity. Until colistin is formed, colistimethate sodium by itself has been shown to display little to no antibacterial activity and is considered an inactive prodrug of colistin.[8]
Colistimethate sodium is eliminated mainly by the renal route, with a fraction of the dose being converted to active colistin in vivo. Colistin undergoes extensive renal tubular reabsorption and therefore is mainly cleared by nonrenal mechanisms.[9, 10] The mechanism behind colistin's bactericidal ability is thought to be indistinguishable from that of polymyxin B, the standard of the polymyxins.[11] Colistin is polycationic and has both hydrophilic and lipophilic moieties. These interact electrostatically with the outer membrane of gram-negative bacteria and competitively displace divalent cations from the membrane lipids, specifically calcium and magnesium.[12] This disrupts the outer membrane and releases lipopolysaccharides.[13] Change in the permeability of the bacterial membrane leads to leakage of the cell content and subsequently cell lysis and death.[2–4] Colistin also has the ability to bind and neutralize the lipopolysaccharide molecule of bacteria, giving it antiendotoxin activity.[2] Colistin has a narrow antibacterial spectrum of activity, with susceptibility mostly against common gram-negative isolates. Most significantly, it displays in vitro activity against MDR gram-negative pathogens such as A. baumannii, P.aeruginosa, and K. pneumoniae. Colistin also has activity against other isolates, such as Enterobacteriaceae,Stenotrophomonas maltophilia,Escherichia coli, Salmonella species, Shigella species, Haemophilus influenzae, Bordetellapertussis, and Legionella pneumophila.[2]
Reports of Colistin Resistance
As mentioned earlier, in the case of MDR gramnegative organisms such as A. baumannii, P.aeruginosa, and K. pneumoniae, the need for alternative treatments has led to the reemergence of colistin. Although colistin has been shown to be effective for the treatment of a wide variety of infections,[3, 14] its use for treating infections caused by these three gram-negative organisms has been impeded by occurrences of colistin resistance. Development of resistance to colistin is a serious concern. As colistin is the last line of defense against these virulent pathogens, resistance to this antibiotic may have devastating effects if no other treatment options are available to combat the infection. Cases of colistin resistance, as well as the mechanisms behind its development, are discussed in the following sections.
Acinetobacter baumannii
The increasing prevalence of A. baumannii infections coupled with its escalating resistance to available treatments and the lack of drug development to cover this pathogen has made it one of the most difficult gram-negative infections to treat and control.[1, 15] The Clinical and Laboratory Standards Institute susceptibility breakpoint for A. baumannii is 2 mg/L or lower, and the resistance breakpoint is 8 mg/L or higher.[16] Although colistin is often considered a reliable agent to treat A. baumannii, reports of resistant strains to this antibiotic are on the rise.[17–20] Recent studies have shown varying rates of resistance as well as the occurrence of hetero-resistant strains (Table 1).[15, 17–29]
In one study, 265 strains of Acinetobacter were collected from two Korean hospitals.[18] Of those 265 isolates, 214 (81%) were determined to be A.baumannii. With use of the broth microdilution method, 27.9% of these isolates were found to be resistant to colistin (minimum inhibitory concentration [MIC] > 16 mg/L) with a minimum concentration required to inhibit 90% of bacteria (MIC90) of 32 mg/L. The A. baumannii strains were further classified into three subgroups based on phylogenetic clustering. In subgroups II and III, most isolates were colistin resistant (64.8% and 88.9%, respectively) but typically remained susceptible to other conventional antibiotics (carbapenems, β-lactams, and ciprofloxacin). In contrast, the A. baumannii isolates in subgroup I had a much lower rate of resistance to colistin (7%), with an increased resistance profile to other antimicrobials.
Similar results were seen in a second study, which compared 17 colistin-resistant isolates of A. baumannii to 17 susceptible strains.[21] Like the previous study, this study also used the broth microdilution method for MICs. Generally, the strains resistant to colistin had increased susceptibility to conventional antibiotics, with substantial decreases in the MICs (up to 16 times lower). The colistin-resistant strains were also found to have a decreased ability to form biofilms, which is associated with diminished antibiotic susceptibility in A. baumannii.[21]
In several studies, colistin also exhibited heteroresistance.[15, 19, 22, 23] Heteroresistance occurs when subpopulations within the strain exhibit reduced susceptibility although the overall MIC is not altered. This makes detection of resistant subpopulations impossible with MIC alone. In one study of 16 A. baumannii isolates, all 16 strains were initially susceptible to colistin with an MIC of 2 mg/L or lower.[15] However, when concentrations of colistin up to 32 times MIC were used, significant regrowth of A.baumannii was noted at 24 hours. By the conclusion of the study, 15 of 16 isolates exhibited heteroresistance. Other studies had similar outcomes, with some heteroresistant strains showing regrowth within 6 hours, regardless of the dosing schedule initiated.[30–32]
A recent study of clinical isolates from the Western Pacific region showed 1 (3.3%) of 30 isolates to be resistant to colistin and 7 isolates (23%) to be colistin heteroresistant.[19] Although lower than that of previously reported cases of colistin resistance or heteroresistance, these findings still emphasize not only the need for adequate dosing, but also the potential use of combination therapy to eradicate these resistant subpopulations.
Pseudomonas aeruginosa
The high mortality rate associated with P.aeruginosa is in part related to its multiple mechanisms of resistance, with some clinical isolates showing panresistance to all United States Food and Drug Administration–approved antibiotics.[1] It has a Clinical and Laboratory Standards Institute susceptibility breakpoint of 2 mg/L or lower, and a resistance breakpoint of 4 mg/L or higher. Infections and resistance due to P. aeruginosa is of even more concern in patients with cystic fibrosis, as it is the most common colonizing pathogen in the lungs and has higher rates of resistance in this population.[25] Although colistin is usually regarded as salvage therapy and is sometimes the only therapeutic option to treat P. aeruginosa, cases of isolates resistant to colistin have emerged (Table 1).
In 385 strains of P. aeruginosa isolates from 57 adults with cystic fibrosis, only 34.9% of nonmucoid and 51.9% of mucoid strains were susceptible to colistin (MIC < 0.5 mg/L).[25] Furthermore, the MIC distribution pattern in this study showed two populations of MICs, which may be indicative of emerging resistance.
A second study of 23 clinical isolates from patients with cystic fibrosis found 11 of these strains to be resistant to colistin, with MICs exceeding 128 mg/L.[24] Also, cases of colistinresistant P. aeruginosa were seen in six children with cystic fibrosis after they received aerosolized colistin for a mean duration of 3.1 years.[26] This rise in colistin resistance by P. aeruginosa is beginning to surface in the cystic fibrosis population, possibly secondary to the widespread use of inhaled colistin in these patients. Because P. aeruginosa plays a large role in the lung destruction and eventual respiratory failure seen with cystic fibrosis, continued development of resistance would be detrimental.
Klebsiella pneumoniae
The need for alternative antimicrobials to treat K. pneumoniae has risen with the increased prevalence of K. pneumoniae carbapenemases-, extended-spectrum β-lactamase (ESBL)-, and metallo-β-lactamase (MBL)-producing strains of this bacteria. Although reports of colistin resistance with this pathogen are sparse, they are significant, as current and future treatment options for ESBL- and MBL-producing K.pneumoniae are limited.
In one study, 18 colistin-resistant (MIC > 8 mg/L) K. pneumoniae isolates were obtained from 13 patients over a 16-month period in an intensive care unit in Greece.[27] All of these patients had a long duration of both colistin treatment (median 27 days) and hospitalization (median 69 days), which likely contributed to the development of resistance. Most recently, 6.8% (15 of 221 isolates)[29] and 27.3% (6 of 22 isolates)[28] of collected K. pneumoniae isolates were found to be colistin resistant in South Korea and Australia, respectively.
Mechanisms of Resistance
Resistance to colistin can develop through adaptive or mutational mechanisms, with almost complete cross-resistance existing between colistin and other polymyxins.[4] A variety of gene mutations cause resistance to colistin by altering the outer membrane of gram-negative bacteria, which is colistin's site of action. Although data on the precise mechanism of resistance are scant and appear to be dependent on specific bacteria, the PmrA-PmrB and PhoP-PhoQ regulatory systems play important roles in its development (Table 2).[33–49] Two-component regulatory systems, such as PmrA-PmrB and PhoP-PhoQ, allow bacteria to respond to environmental changes by modifying the expression of genes. When these regulatory systems interact with one another, they have been shown to have even more profound effects.[33]
One mechanism of resistance involves changes in the structure of the bacteria's negatively charged surface lipopolysaccharides and lipid A. These modifications occur as a result of the activation of the PmrA-PmrB system, which is regulated by the PhoP-PhoQ system, but can also act independently in mildly acidic conditions or with high concentrations of iron.[34, 35] The PmrAPmrB system regulates two loci, PmrE and PmrHFIJKLM, which are responsible for the changes in lipid A and are essential for polymyxin resistance.[34, 36–40] When activated, the PmrA-PmrB system adds ethanolamine to the phosphate groups of the lipopolysaccharides and lipid A and also inserts aminoarabinose at the 4' phosphate of lipid A.[34, 35, 41, 42] These changes lower the overall charge of the lipopolysaccharide, thereby reducing the binding affinity of the cationic polymyxins.[43]
Environmental pH and magnesium concentrations are two environmental factors that appear to greatly affect the expression of the bacteria's genes and the subsequent development of resistance. In one study of Salmonella enterica grown in 10 mM magnesium chloride at a pH of 5.8, the organisms were approximately 100,000 times more resistant to polymyxin B than strains grown at a pH of 7.7.[41] This increase is attributed to increased activation of the PmrA-PmrB system at slightly acidic pH values and micromolar magnesium concentrations. It may be possible to monitor or correct pH and magnesium levels in order to help prevent resistance due to these environmental factors, but more information is needed before this can be determined. Because studies examining environmental pH and magnesium compared with the rate of resistance are limited, it remains unclear what steps clinicians should take. It is clear, however, that the effects of pH and magnesium require an active PmrA-PmrB system. The PmrA null mutants have failed to exhibit polymyxin resistance.[44]
Low magnesium concentrations also lead to the development of resistance by activating PhoP and PmrA, which not only modifies the bacteria's lipopolysaccharides but also increases the expression of a gene that has been shown to be a major factor in the development of resistance—the OprH gene.[33, 35, 37, 44] The OprH, PhoP, and PhoQ genes form an operon that is controlled by both PhoP and magnesium concentrations and contributes to polymyxin resistance.[35, 45] The OprH gene, which lies immediately downstream from the PhoP-PhoQ regulatory system, encodes an outer membrane protein, OprH, that has enhanced expression in low magnesium level conditions.[36] These OprH proteins occupy membrane magnesium sites and reduce the binding sites for colistin, therefore contributing to resistance.[35, 45–47]
The presence of exogenous polyamines (spermidine, spermine, putrescine, and cadaverine) has also been shown to induce the expression of the OprH-PhoP-PhoQ operon, resulting in increased MICs of not only polymyxins, but also aminoglycosides, quinolones, and fluorescent dyes against P.aeruginosa, regardless of the presence of cations.[48] Although OprH is presumed to play a role in resistance, it has been proven that its presence is not necessary for resistance to occur, since OprHdeficient strains of P. aeruginosa remain polymyxin resistant.[33] Similarly, studies have shown that although PhoP is essential for the transcription of the OprH-PhoP-PhoQ operon, PhoP-null strains of P. aeruginosa retain polymyxin resistance.[45] This is significant as it exemplifies the independent role that the PmrAPmrB system plays in polymyxin resistance, as well as the potential for other unidentified mechanisms of resistance.
Recently, the morphology and topography of colistin-resistant bacteria have been found to differ from that of colistin-susceptible cells, which could give us further insight into the genetic mechanisms leading to colistin resistance.[49] An atomic force microscopy study was performed of both colistin-resistant and colistin-susceptible strains at different growth phases.[49] Compared with spherically shaped colistin-resistant bacteria at early and mid-logarithmic phases, susceptible cells were found to be rod shaped with pili present at all phases. The number and length of pili for colistin-resistant cells were greatly reduced, which the authors note could be the reason colistinresistant cells are unable to form a biofilm. In addition, colistin-resistant cells had a greater topographic variability and finer surface texture. In the stationary phase, elongated worm-like cells were more prevalent in the susceptible group versus the resistant group, which showed more heterogeneity among the cells in this phase. Of interest, levels of bacterial outer membrane damage after treatment with colistin were similar for both susceptible and resistant cells, showing the ability of colistin-resistant cells to maintain interaction with the outer membrane.[21, 49] Based on these findings, it is evident that specific studies examining the genetic mechanisms behind these morphologic and topographic differences need to be performed, so that we may better understand the resistance associated with colistin.
Toxicity
Early use of colistin was linked to multiple reports of nephrotoxicity and neurotoxicity.[2] It was from this fear of toxicity that its use was halted shortly thereafter.[2–4, 50–52] Reports over the past decade, however, have shown that the toxicity associated with the polymyxins is much less than originally believed.[5, 50, 53–56] The results from earlier reports were most likely a result of a lack of pharmacokinetic, pharmacodynamic, and toxicity studies.[2] Also, incorrect dosing, which may have resulted directly from confusing dosage forms and units, and the presence of other nephrotoxic drugs or conditions may have contributed to these toxicities.[57] Several studies have since examined the safety of colistin, and their results give us further insight into the toxicities associated with the antibiotic (Table 3).[51, 53–56, 58–62]
Nephrotoxicity
Several studies have proven satisfactory safety profiles with intravenous colistimethate sodium 160 mg 3 times/day in patients with normal renal function.[58, 63, 64] The authors of two studies found that no serious adverse effects occurred with this dosage regimen in the cystic fibrosis populations that they studied, and that there were no notable changes in renal function.[58, 63] Furthermore, colistin has recently been found to have a more favorable toxicity profile compared with the aminoglycosides, which were originally used in place of colistin because of their suspected decrease in toxic effects.[55, 65] Another group found an observable decrease in renal function in patients receiving aminoglycosides, which was further worsened by coadministration of colistin.[55] Colistin used as monotherapy or in combination with nonnephrotoxic antibiotics, however, did not appear to cause renal damage. Two additional studies concluded that colistin was generally well tolerated in critically ill patients.[53, 59] In one of the studies,[59] serum creatinine level slightly increased by 0.25 mg/dl from baseline during treatment, but it is important to note that the study was performed in patients with decreased renal function.
Although this increase in creatinine level is of concern, no serious adverse effects occurred, and there were no data suggesting renal toxicity. Although the nephrotoxicity associated with colistin is not as toxic as originally thought, it is still an adverse effect that must be considered when administering the antibiotic. A few recent studies have given us more insight into colistin-induced renal impairment. One group of authors examined the occurrence of acute renal failure with use of the RIFLE—risk, injury, failure, loss, and end-stage kidney disease—criteria, by completing a retrospective review of patients (aged ≥ 18 yrs) who received intravenous colistimethate sodium (≥ 72 hrs) between January 2003 and December 2007.[60] Among the 66 patients, they found that the peak serum creatinine level during colistimethate sodium treatment met the RIFLE criteria for nephrotoxicity in 45% of patients, and that 21% of patients stopped colistimethate sodium therapy due to nephrotoxicity. The probability of renal toxicity increased in proportion to the overall total colistimethate sodium dose, and patients who received colistimethate sodium for longer than 14 days were 3.7 times more likely to experience nephrotoxicity. These findings are consistent with those of another group, who similarly found the change in serum creatinine level to be correlated with the cumulative dose of colistimethate sodium administered in their prospective cohort study.[61] Also, in the first study mentioned, the authors found that serum creatinine levels returned to baseline within 1 month after cessation of colistin,[60] suggesting reversibility of nephrotoxicity after discon- tinuation. A third group of authors performed a case-control study to evaluate the occurrence of nephrotoxicity and to analyze the characteristics and risk factors of patients who develop nephrotoxicity. [62] Hypoalbuminemia and concomitant use of a nonsteroidal antiinflammatory drug were the only statistically significant independent risk factors.
Neurotoxicity
Adverse effects such as paresthesias, visual alterations, ataxia, and neuromuscular blockade are possible with polymyxins as a class. These neurologic effects, however, are usually reversible after the cessation of treatment and usually occur in patients receiving prolonged treatment. Cases of neurotoxicity due to colistin have been mild and rare, with the current rate of neurotoxicity estimated to be 0–7%.[66] Little to no data exist on colistin alone causing neurotoxicity in patients. One study showed that in 21 patients treated with colistin for ventilator-associated pneumonia, there were no reports of neuromuscular blockade after patients were evaluated with an electro-physiologic study to detect the presence of neuromuscular transmission blockade and critical illness polyneuropathy.[67] Colistin was concluded to be a safe treatment alternative. Another study that included 17 patients who received colistin for more than 4 weeks of treatment, found that one patient appeared to develop neuropathy believed to be caused by colistin.[59] The authors concluded that despite this one finding, colistin was a safe and efficacious alternative therapy.
Summary
The toxicities associated with colistin have been found to be better correlated with the total cumulative amount of colistimethate sodium administered versus single or daily doses, and may occur more frequently in patients with hypoalbuminemia and concurrent nonsteroidal antiinflammatory drug use. Taking these risk factors into consideration when dosing can help to prevent adverse events. Although recent studies have found colistin-induced serum creatinine level increases, this adverse effect has been found to be reversible. Overall, many studies have shown that colistin is generally well tolerated, with less nephrotoxicity and neurotoxicity than was once thought.
Optimization of Colistin Dosing
Lack of Universal Dose Unit
We refer the reader to excellent review articles that extensively address this contemporary issue.[2, 5, 68] The limited data on colistin's pharmacokinetic and pharmacodynamic properties create immense confusion in assessing optimal dosing regimens that maximize antibacterial activity and minimize toxicity.[2, 5] Before attempting to determine optimal dosages for colistin, a universal dose unit measurement is needed when referring to the amount of drug being administered. The uncertainty related to dosing colistin is because some products use milligrams, whereas others use international units (IU). It has been established that there are approximately 12,500 IU per 1 mg of colistimethate sodium.[5] For example an average dose of colistimethate sodium is 2 million IU, which corresponds to 160 mg of drug.
To add further confusion, some products use milligrams of "colistin base activity" rather than milligrams of colistimethate sodium. It must be emphasized again that colistimethate sodium and colistin cannot be used interchangeably, especially when dosing. There are approximately 2.67 mg of colistimethate sodium per 1 milligram of colistin base.[5] To continue the example above, 2 million IU equals 160 mg of colistimethate sodium, which is equivalent to approximately 60 mg of colistin base. As can be seen by this complexity, the use of a unified dosage form and unit would greatly benefit the discussion of colistin dosing.
Discrepancies between Recommended Dosage Regimens
Once the proper dosage form and unit are established, the optimal dosage for patients must be decided. Because colistin is an older drug, there is little to no information on pharmacokinetics, pharmacodynamics, and toxicity to establish a safe and effective dosage regimen. Therefore, current dosage regimens are primarily derived from manufacturers' package inserts. The manufacturer of Colomycin (Xellia Pharmaceuticals, Copenhagen, Denmark) recommends that patients weighing more than 60 kg receive 1–2 million IU 3 times/day, equivalent to colistimethate sodium 80–160 mg 3 times/day, with a recommended daily upper limit of 6 million IU, or 480 mg of colistimethate sodium.[69] The manufacturer of Coly-Mycin M (Parkedale Pharmaceuticals, Inc., Rochester, MN) recommends 2.5–5 mg/kg/day colistin base activity in 2–4 divided doses, equivalent to colistimethate sodium 6.67–13.3 mg/kg/day or 83,375–166,250 IU/kg/day.[70] The recommended maximum daily dose of colistin is 10 million IU, or 800 mg of colistimethate sodium, from the manufacturer of Coly-Mycin M, which is approximately double the recommended daily dose from the manufacturer of Colomycin. Table 4 provides a comparison of the two products.
It should be of concern that the manufacturer of Coly-Mycin M recommends approximately double the dose of that recommended by the manufacturer of Colomycin. This lack of uniformity between manufacturers could lead to underdosing, inevitably leading to treatment failure and development of resistance.[68] More information is needed before we can begin to determine which regimens provide the best outcomes with acceptable safety.
Use of Pharmacodynamics to Guide Optimal Dosing
To fully optimize regimen selection of colistimethate sodium and colistin, it is important to appreciate the pharmacodynamics of colistin. Colistin is a rapidly bactericidal antimicrobial that possesses a significant postantibiotic effect against P. aeruginosa, A. baumannii, and K.pneumoniae. [24] The colistin area under the concentration-time curve (AUC):MIC ratio has been found to be the parameter best associated with efficacy.[71, 72] Researchers used neutropenic murine thigh and lung models to determine the pharmacokinetic-pharmacodynamic index of colistin that best correlates with efficacy against P.aeruginosa, and to determine the index target values needed for specific antibacterial effects.[71] For both the thigh and lung models, the unbound AUC:MIC (fAUC:MIC) ratio was the pharmacokinetic-pharmacodynamic index that had the strongest relationship to bacterial burden, with R2 values equaling 87% and 89% for the two models, respectively. The time where free drug concentration above the MIC (fT>MIC) was also closely correlated with efficacy, with R2 values equaling 84% and 88% for the thigh and lung models, respectively. The researchers noted, however, that fAUC:MIC ratio is still the pharmacokinetic-pharmacodynamic index most likely to be associated with colistin's activity because the scatter for the fT>MIC was relatively large in the 20–30% range, and because concentration-dependent killing has been seen with colistin in vitro. In the lung infection model, fAUC:MIC ratio target ranges for the three different strains of bacteria were 15.6–22.7, 27.6–36.1, and 53.3–66.7 for 1-log, 2-log, and 3- log bacterial kill, respectively. For the thigh infection model, target values ranged from 12.2–16.7, 36.9–45.9, and 105–141 for 1-log, 2-log, and 3-log kills, respectively. Although pharmacodynamic similarities among the two different sites of infection were seen, the observed differences emphasize the dosing alterations that may be needed based on site and type of infection.
Particularly in severe infections such as endocarditis, infections of prostheses, and ventilator-associated pneumonia, a high density of bacteria is known to exist, which may impact colistin pharmacodynamics.[73] Recently, the antibacterial activity of colistin was shown to be attenuated when facing a higher bacterial density. Investigators determined the extent and rate of killing by colistin to be greatly decreased at high compared with low inocula.[73] Against a genetically characterized clinical isolate of P. aeruginosa(PAO1), colistin killing was 23-fold slower at 109 and 6-fold slower at 108 compared with 106 colonyforming units, and 32-fold higher concentrations were required at 109 versus 106 colony-forming units. Although animal and in vivo studies are needed to further assess this inoculum effect, this study highlights the fact that higher colistin doses may be needed to treat sequestered, deep-seated infections with high bacterial densities.
When optimizing regimens for colistin, frequency of dosing is another important aspect to determine. One group of researchers evaluated the antibacterial activity and emergence of resistance that occurred with three different dosing intervals: 8, 12, and 24 hours.[72] Three different dosage regimens were used: 0.23 mg every 8 hours (0.30-mg loading dose), 0.39 mg every 12 hours (0.45-mg loading dose), and 0.89 mg every 24 hours (0.90-mg loading dose). The every-8-hour regimen simulated the expected maximum concentration ([Cmax] 3 mg/L) and minimum concentration (0.75 mg/L) at steady-state concentrations when colistin is given according to the manufacturer's recommendations. The every-12-hour and every-24-hour regimens were designed to provide higher target Cmax values (4.5 and 9.0 mg/L, respectively). The researchers found overall bacterial killing and regrowth to be similar among the three regimens. However, emergence of resistance increased as dosing interval increased, and the 8-hour regimen was the most effective at minimizing resistance. Concentrations remained above the MIC for approximately 80%, 72%, and 53% of the 72-hour treatment period for the 8-, 12-, and 24-hour dosage regimens, respectively. As colistin resistance continues to rise, these findings are important to keep in mind. In addition, other infection-specific considerations that can alter pharmacodynamics must be taken into account when dosing colistin and colistimethate sodium.
Dosing in Critically Ill Patients
In critically ill patients with multiorgan dysfunction and severe infections due to MDR organisms, treatment options are especially limited. Colistin remains a necessary last-line option for these patients. What is most concerning, however, is the lack of clinical guidelines and the presence of unclear dosing recommendations in this patient population.
Recent evidence has shown that the pharmacokinetics of colistimethate sodium and colistin in critically ill patients differ from those previously found among patients with cystic fibrosis.[68] In critically ill patients, who may have multiorgan failure, sepsis, or a wider range of renal impairment, the differences are important to take into account. Although the half-life of colistin is approximately 4 hours in patients with cystic fibrosis, it is longer in critically ill patients.[68] The half-life is 14.4 hours in critically ill patients, and the rate of formation of colistin from colistimethate sodium is different from previously published data.[74] In addition, larger volumes of distribution and lower concentrations of the antibiotic have been seen in critically ill patients with sepsis.[75] These differences have the ability to impact the effects of colistimethate sodium and colistin, which could require alterations in the dosage regimen. Although pharmacokinetic and pharmacodynamic data in this patient population are scarce, some studies have given us further insight into changes that may be needed when dosing critically ill patients.
One group of authors completed a population analysis to examine the pharmacokinetics of colistin after the administration of intravenous doses of colistimethate sodium in critically ill patients.[74] Patients received 3 million IU (240 mg) of colistimethate sodium intravenously every 8 hours or 160 mg every 8 hours if creatinine clearance was less than 50 ml/minute. The predicted plasma Cmax was 0.60 mg/L after the first dose and 2.3 mg/L at steady state. The authors found that after the first few doses of the regimen, colistin concentrations were below the Clinical and Laboratory Standards Institute MIC breakpoint of 2 mg/L for P. aeruginosa and Enterobacteriaceae. In addition, at steady state, plasma concentrations were below the MIC breakpoints for many of the cases. These results are of particular concern in critically ill patients, for whom a delay in appropriate treatment or suboptimal efficacy of the current regimen can lead to resistance and ultimately increased mortality. The authors speculated that a loading dose of colistimethate sodium is warranted. At 3 million IU every 8 hours, it would take 2–3 days before the steady-state concentration is achieved. Thus, the authors suggest that a colistimethate sodium loading dose of 9 or 12 million IU along with a 4.5 million IU maintenance dose every 12 concentration at a faster rate and with less frequent administration.
Similarly, another group assessed the steady-state serum concentrations of colistin after intravenous administration of colistimethate sodium 225 mg every 8 hours in 14 patients.[75] The average Cmax was found to be 2.93 mg/L, which the authors noted would most likely lead to suboptimal Cmax:MIC ratios for strains with higher MICs (e.g., A. baumannii and P.aeruginosa). The researchers concluded that higher doses of colistimethate sodium be considered. Based on these two studies, it is evident that further investigations using higher colistimethate sodium doses must be performed in critically ill patients to determine whether there is improved efficacy without increased toxicity.
The pharmacokinetic parameters of colistimethate sodium and colistin were examined in patients with stage 5 kidney disease or severe liver disease compared with healthy subjects.[76] Clearance of colistimethate sodium was found to be lower in the group of patients with kidney disease, and Cmax, half-life, and AUC were higher. In addition, conversion of colistimethate sodium to colistin and overall colistin exposure were increased in these patients, and clearance of colistin was decreased. Potentially, these results would have led to the neurotoxicity that occurred in the kidney disease group, as 3 of 10 patients in this group experienced paresthesias (which resolved in 24–48 hrs), compared with no patients in the liver disease group. Previously, a regimen of 2.5 mg/kg every 48 hours in patients receiving renal replacement therapy was suggested,[77] but this regimen has been found to be inadequate in some cases.[78] This study in particular highlights the fact that dosing may need to be altered in patients with renal failure.
In critically ill patients, for whom colistin's half-life appears to be longer, the potential for a longer dosing interval may be an option. Some studies, however, have found that as the interval between colistin doses becomes more extended, the prevalence of resistance increases.[72] This potentially serious consequence should be considered when deciding whether or not to use extended-interval dosing.
Overall, these data suggest that colistin pharmacokinetics are severely altered in critically ill patients. To maximize the AUC:MIC ratio, the predictive pharmacodynamic parameter of colistin, higher doses of colistimethate sodium and alterations in the dosing interval may be warranted. Because of colistin's toxicities, however, these may not be achievable. In these instances, combination therapy should be considered for optimal therapy and prevention of resistance.
Conclusion
Colistin has proved to be an important alternative for MDR gram-negative infections. However, reports of colistin-resistant strains have created a potentially dangerous scenario since it is the last line of defense. Colistin resistance is largely attributed to the PmrA-PmrB and PhoP-PhoQ regulatory systems and their responses to environmental changes. The activation of the PmrA-PmrB and PhoP-PhoQ regulatory systems produces resistance by activating a variety of genes that lower the negative charge of the outer membrane and decrease the number of binding sites for the cationic polymyxins. Although studies have begun to reveal the mechanisms behind colistin resistance, further research is needed to fully understand the impact that the two regulatory systems have on resistance, as well as the dosages of colistin needed to inhibit and overcome these developing patterns.
The development of colistin resistance has also been linked to inadequate dosing. This highlights the importance of dose optimization, especially in critically ill patients with MDR bacterial infections. Although higher doses appear beneficial, the lack of pharmacodynamic and pharmacokinetic data regarding colistin makes determination of appropriate dosing difficult. Colistin remains an essential alternative for most MDR gram-negative infections; however, cases of resistant strains should be a cause of much concern. Therefore, newer agents and colistin combination therapy are avenues that should be considered to optimize therapeutic regimens in the fight against evolving and highly resistant gram-negative infections.
References
1. Talbot GH, Bradley J, Edwards JE Jr, Gilbert D, Scheld M, Bartlett JG. Bad bugs need drugs: an update on the development pipeline from the antimicrobial availability task force of the Infectious Diseases Society of America. Clin Infect Dis 2006;42:657–68. (Erratum in Clin Infect Dis 2006;42: 1065.
2. Li J, Nation RL, Milne RW, Turnidge JD, Coulthard K. Evaluation of colistin as an agent against multi-resistant gram-negative bacteria. Int J Antimicrob Agents 2005;25:11–25.
3. Falagas ME, Kasiakou SK, Tsiodras S, Michalopoulos A. The use of intravenous and aerosolized polymyxins for the treatment of infections in critically ill patients: a review of the recent literature. Clin Med Res 2006;4:138–46.
4. Falagas ME, Kasiakou SK. Colistin: the revival of polymyxins for the management of multidrug-resistant gram-negative bacterial infections. Clin Infect Dis 2005;40:1333–41.
5. Li J, Nation RL, Turnidge JD, et al. Colistin: the re-emerging antibiotic for multidrug-resistant gram-negative bacterial infections. Lancet Infect Dis 2006;6:589–601.
6. Li J, Coulthard K, Milne R, et al. Steady-state pharmacokinetics of intravenous colistin methanesulphonate in patients with cystic fibrosis. J Antimicrob Chemother 2003;52:987–92.
7. Li J, Milne RW, Nation RL, Turnidge JD, Coulthard K. Stability of colistin and colistin methanesulfonate in aqueous media and plasma as determined by high-performance liquid chromatography. Antimicrob Agents Chemother 2003;47: 1364–70.
8. Bergen PJ, Li J, Rayner CR, Nation RL. Colistin methanesulfonate is an inactive prodrug of colistin against Pseudomonasaeruginosa. Antimicrob Agents Chemother 2006;50:1953–8.
9. Rodriguez CH, Pautaso J, Bombicino K, Vay C, Famiglietti A. Sensitivity to colistin: evaluation of cut-off points available in disk diffusion test [in Spanish]. Rev Argent Microbiol 2004;36:125–9.
10. Li J, Milne RW, Nation RL, Turnidge JD, Smeaton TC, Coulthard K. Use of high-performance liquid chromatography to study the pharmacokinetics of colistin sulfate in rats following intravenous administration. Antimicrob Agents Chemother 2003;47:1766–70.
11. Storm DR, Rosenthal KS, Swanson PE. Polymyxin and related peptide antibiotics. Annu Rev Biochem 1977;46:723–63.
12. Dixon RA, Chopra I. Leakage of periplasmic proteins from Escherichia coli mediated by polymyxin B nonapeptide. Antimicrob Agents Chemother 1986;29:781–8.
13. Peterson AA, Hancock RE, McGroarty EJ. Binding of polycationic antibiotics and polyamines to lipopolysaccharides of Pseudomonas aeruginosa. J Bacteriol 1985;164:1256–61.
14. Kasiakou SK, Rafailidis PI, Liaropoulos K, Falagas ME. Cure of post-traumatic recurrent multiresistant gram-negative rod meningitis with intraventricular colistin. J Infect 2005;50: 348–52.
15. Li J, Rayner CR, Nation RL, et al. Heteroresistance to colistin in multidrug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 2006;50:2946–50.
16. Galani I, Kontopidou F, Souli M, et al. Colistin susceptibility testing by Etest and disk diffusion methods. Int J Antimicrob Agents 2008;31:434–9.
17. Arroyo LA, Garcia-Curiel A, Pachon-Ibanez ME, et al. Reliability of the E-test method for detection of colistin resistance in clinical isolates of Acinetobacter baumannii. J Clin Microbiol 2005;43:903–5.
18. Ko KS, Suh JY, Kwon KT, et al. High rates of resistance to colistin and polymyxin B in subgroups of Acinetobacterbaumannii isolates from Korea. J Antimicrob Chemother 2007;60:1163–7.
19. Yau W, Owen RJ, Poudyal A, et al. Colistin heteroresistance in multidrug-resistant Acinetobacter baumannii clinical isolates from the western pacific region in the SENTRY antimicrobial surveillance programme. J Infect 2009;58:138–44.
20. Doi Y, Husain S, Potoski B, et al. Extensively drug-resistant Acinetobacter baumannii. Emerg Infect Dis 2009;15:980–2.
21. Li J, Nation RL, Owen RJ, Wong S, Spelman D, Franklin C. Antibiograms of multidrug-resistant clinical Acinetobacterbaumannii: promising therapeutic options for treatment of infection with colistin-resistant strains. Clin Infect Dis 2007;45:594–8.
22. Hawley JS, Murray CK, Griffith ME, et al. Susceptibility of Acinetobacter strains isolated from deployed U.S. military personnel. Antimicrob Agents Chemother 2007;51:376–8.
23. Hernan RC, Karina B, Gabriela G, Marcela N, Carlos V, Angela F. Selection of colistin-resistant Acinetobacter baumannii isolates in postneurosurgical meningitis in an intensive care unit with high presence of heteroresistance to colistin. Diagn Microbiol Infect Dis 2009;65:188–91.
24. Li J, Turnidge J, Milne R, Nation RL, Coulthard K. In vitro pharmacodynamic properties of colistin and colistin methanesulfonate against Pseudomonas aeruginosa isolates from patients with cystic fibrosis. Antimicrob Agents Chemother 2001;45:781–5.
25. Schulin T. In vitro activity of the aerosolized agents colistin and tobramycin and five intravenous agents against Pseudomonasaeruginosa isolated from cystic fibrosis patients in southwestern Germany. J Antimicrob Chemother 2002;49:403–6.
26. Denton M, Kerr K, Mooney L, et al. Transmission of colistin-resistant Pseudomonas aeruginosa between patients attending a pediatric cystic fibrosis center. Pediatr Pulmonol 2002;34: 257–61.
27. Antoniadou A, Kontopidou F, Poulakou G, et al. Colistin-resistant isolates of Klebsiella pneumoniae emerging in intensive care unit patients: first report of a multiclonal cluster. J Antimicrob Chemother 2007;59:786–90.
28. Poudyal A, Howden B, Bell J, et al. In vitro pharmacodynamics of colistin against multidrug-resistant Klebsiella pneumoniae. J Antimicrob Chemother 2008;62:1311–18.
29. Suh J-Y, Son J, Chung D, et al. Nonclonal emergence of colistin-resistant Klebsiella pneumoniae isolates from blood samples in South Korea. Antimicrob Agents Chemother 2010;54:560–2.
30. Owen RJ, Li J, Nation RL, Spelman D. In vitro pharmaco-dynamics of colistin against Acinetobacter baumannii clinical isolates. J Antimicrob Chemother 2007;59:473–7.
31. Tan TY, Ng LS, Tan E, Huang G. In vitro effect of minocycline and colistin combinations on imipenem-resistant Acinetobacterbaumannii clinical isolates. J Antimicrob Chemother 2007;60:421–3.
32. Tan CH, Li J, Nation RL. Activity of colistin against heteroresistant Acinetobacter baumannii and emergence of resistance in an in vitro pharmacokinetic/pharmacodynamic model. Antimicrob Agents Chemother 2007;51:3413–15.
33. Macfarlane EL, Kwasnicka A, Ochs MM, Hancock RE. PhoP-PhoQ homologues in Pseudomonas aeruginosa regulate expression of the outer-membrane protein OprH and polymyxin B resistance. Mol Microbiol 1999;34:305–16.
34. Gunn JS, Ryan SS, Van Velkinburgh JC, Ernst RK, Miller SI. Genetic and functional analysis of a PmrA-PmrB-regulated locus necessary for lipopolysaccharide modification, antimicrobial peptide resistance, and oral virulence of Salmonella enterica Serovar Typhimurium. Infect Immun 2000;68:6139–46.
35. Soncini FC, Groisman EA. Two-component regulatory systems can interact to process multiple environmental signals. J Bacteriol 1996;27:6796–801.
36. McPhee JB, Bains M, Winsor G, et al. Contribution of the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems to Mg2+-induced gene regulation in Pseudomonasaeruginosa. J Bacteriol 2006;188:3995–4006.
37. McPhee JB, Lewenza S, Hancock RE. Cationic antimicrobial peptides activate a two-component regulatory system, PmrAPmrB, that regulates resistance to polymyxin B and cationic antimicrobial peptides in Pseudomonas aeruginosa. Mol Microbiol 2003;50:205–17.
38. Moskowitz SM, Ernst RK, Miller SI. PmrAB, a two-component regulatory system of Pseudomonas aeruginosa that modulates resistance to cationic antimicrobial peptides and addition of aminoarabinose to lipid A. J Bacteriol 2004;186:575–9.
39. Trent MS, Ribeiro AA, Lin S, Cotter RJ, Raetz CR. An inner membrane enzyme in Salmonella and Escherichia coli that transfers 4-amino-4-deoxy-L-arabinose to lipid A: induction on polymyxin-resistant mutants and role of a novel lipid-linked donor. J Biol Chem 2001;276:43122–31.
40. Zhang L, Dhillon P, Yan H, Farmer S, Hancock RE. Interactions of bacterial cationic peptide antibiotics with outer and cytoplasmic membranes of Pseudomonas aeruginosa. Antimicrob Agents Chemother 2000;44:3317–21.
41. Perez JC, Groisman EA. Acid pH activation of the PmrA/PmrB two-component regulatory system of Salmonella enterica. Mol Microbiol 2007;63:283–93.
42. Gunn JS, Miller SI. PhoP-PhoQ activates transcription of pmrAB, encoding a two-component regulatory system involved in Salmonella typhimurium antimicrobial peptide resistance. J Bacteriol 1996;178:6857–64.
43. Gunn JS, Lim KB, Krueger J, et al. PmrA-PmrB-regulated genes necessary for 4-aminoarabinose lipid A modification and polymyxin resistance. Mol Microbiol 1998;27:1171–82.
44. Groisman EA, Kayser J, Soncini FC. Regulation of polymyxin resistance and adaptation to low-Mg2+ environments. J Bacteriol 1997;179:7040–5.
45. Macfarlane EL, Kwasnicka A, Hancock RE. Role of Pseudomonas aeruginosa PhoP-phoQ in resistance to antimicrobial cationic peptides and aminoglycosides. Microbiology 2000;146(pt 10):2543–54.
46. Nicas TI, Hancock RE. Alteration of susceptibility to EDTA, polymyxin B and gentamicin in Pseudomonas aeruginosa by divalent cation regulation of outer membrane protein H1. J Gen Microbiol 1983;129:509–17.
47. Nicas TI, Hancock RE. Outer membrane protein H1 of Pseudomonas aeruginosa: involvement in adaptive and mutational resistance to ethylenediaminetetraacetate, polymyxin B, and gentamicin. J Bacteriol 1980;143:872–8.
48. Kwon DH, Lu CD. Polyamines induce resistance to cationic peptide, aminoglycoside, and quinolone antibiotics in Pseudomonas aeruginosa PAO1. Antimicrob Agents Chemother 2006;50:1615–22.
49. Soon RL, Nation RL, Hartley PG, Larson I, Li J. Atomic force microscopy investigation of the morphology and topography of colistin-heteroresistant Acinetobacter baumannii strains as a function of growth phase and in response to colistin treatment. Antimicrob Agents Chemother 2009;53:4979–86.
50. Falagas ME, Kasiakou SK. Toxicity of polymyxins: a systematic review of the evidence from old and recent studies [online exclusive article]. Crit Care 2006;10:R27. Available from http://ccforum.com/content/10/1/R27.
51. Kasiakou SK, Michalopoulos A, Soteriades ES, Samonis G, Sermaides GJ, Falagas ME. Combination therapy with intravenous colistin for management of infections due to multidrug-resistant gram-negative bacteria in patients without cystic fibrosis. Antimicrob Agents Chemother 2005;49: 3136–46.
52. Falagas ME, Bliziotis IA, Kasiakou SK, Samonis G, Athanassopoulou P, Michalopoulos A. Outcome of infections due to pandrug-resistant (PDR) gram-negative bacteria [online exclusive article]. BMC Infect Dis 2005;5:24. Available from http://www.biomedcentral.com/1471-2334/5/24.
53. Reina R, Estenssoro E, Saenz G, et al. Safety and efficacy of colistin in Acinetobacter and Pseudomonas infections: a prospective cohort study. Intensive Care Med 2005;31:1058–65.
54. Cheng C-Y, Sheng W-H, Wang J-T, et al. Safety and efficacy of intravenous colistin (colistin methanesulphonate) for severe multidrug-resistant gram-negative bacterial infections. Int J Antimicrob Agents 2010;25:297–300.
55. Al-Aloul M, Miller H, Alapati S, Stockton PA, Ledson MJ, Walshaw MJ. Renal impairment in cystic fibrosis patients due to repeated intravenous aminoglycoside use. Pediatr Pulmonol 2005;39:15–20.
56. Santamaria C, Mykietiuk A, Temporiti E. Nephrotoxicity associated with the use of intravenous colistin. Scand J Infect Dis 2009;41:767–9.
57. Li J, Rayner CR, Nation RL. Colistin-associated acute renal failure: revisited. South Med J 2005;98:1229–30.
58. Conway SP, Etherington C, Munday J, Goldman MH, Strong JJ, Wootton M. Safety and tolerability of bolus intravenous colistin in acute respiratory exacerbations in adults with cystic fibrosis. Ann Pharmacother 2000;34:1238–42.
59. Falagas ME, Rizos M, Bliziotis IA, Rellos K, Kasiakou SK, Michalopoulos A. Toxicity after prolonged (more than four weeks) administration of intravenous colistin [online exclusive article]. BMC Infect Dis 2005;5:1. Available from http://www.biomedcentral.com/1471-2334/5/1.
60. Hartzell J, Neff R, Ake J. Nephrotoxicity associated with intravenous colistin (colistimethate sodium) treatment at a tertiary care medical center. Clin Infect Dis 2009;48:1724–8.
61. Falgas M, Fragoulis K, Kasiakou S, et al. Nephrotoxicity of intravenous colistin: a prospective evaluation. Int J Antimicrob Agents 2005;26:504–7.
62. Kim J, Lee K-H, Yoo S, et al. Clinical characteristics and risk factors of colistin-induced nephrotoxicity. Int J Antimicrob Agents 2009;34:434–8.
63. Ledson MJ, Gallagher MJ, Cowperthwaite C, Convery RP, Walshaw MJ. Four years' experience of intravenous Colomycin in an adult cystic fibrosis unit. Eur Respir J 1998;12:592–4.
64. Conway SP, Pond MN, Watson A, Etherington C, Robey HL, Goldman MH. Intravenous colistin sulphomethate in acute respiratory exacerbations in adult patients with cystic fibrosis. Thorax 1997;52:987–93.
65. Assadamongkol K, Tapaneya-Olarn W, Chatasingh S. Urinary N-acetyl-beta-D-glucosaminidase (NAG) in aminoglycoside nephrotoxicity. J Med Assoc Thai 1989;72(suppl 1):42–6.
66. Molina J, Cordero E, Pachon J. New Information about the polymyxin/colistin class of antibiotics. Expert Opin Pharmacother 2009;10:2811–28.
67. Garnacho-Montero J, Ortiz-Leyba C, Jimenez-Jimenez FJ, et al. Treatment of multidrug-resistant Acinetobacter baumannii ventilator-associated pneumonia (VAP) with intravenous colistin: a comparison with imipenem-susceptible VAP. Clin Infect Dis 2003;36:1111–18.
68. Nation R, Li J. Colistin in the 21st century. Curr Opin Infect Dis 2009 2009;22:535–43.
69. Forest Laboratories Ltd. Colomycin (colistimethate sodium) injection summary of product characteristics. Bexley, Kent, United Kingdom; 2010.
70. Monarch Pharmaceuticals. Coly-Mycin M (colistimethate) parenteral package insert. Bristol, TN; 2005.
71. Dudhani RV, Turnidge JD, Coulthard K, et al. Elucidation of the pharmacokinetic/pharmacodynamic determinant of colistin activity against Pseudomonas aeruginosa in murine thigh and lung infection models. Antimicrob Agents Chemother 2010;54:1117–24.
72. Bergen PJ, Li J, Nation RL, Turnidge JD, Coulthard K, Milne RW. Comparison of once-, twice- and thrice-daily dosing of colistin on antibacterial effect and emergence of resistance: studies with Pseudomonas aeruginosa in an in vitro pharmaco-dynamic model. J Antimicrob Chemother 2008;61:636–42.
73. Bulitta JB, Yang JC, Yohonn L, et al. Attenuation of colistin bactericidal activity by high inoculum of Pseudomonasaeruginosa characterized by a new mechanism-based population pharmacodynamic model. Antimicrob Agents Chemother 2010;54:2051–62.
74. Plachouras D, Karvanen M, Friberg L, et al. Population pharmacokinetic analysis of colistin methanesulfonate and colistin after intravenous administration in critically ill patients with infections caused by gram-negative bacteria. Antimicrob Agents Chemother 2009;53:3430–6.
75. Markou N, Markantonis S, Dimitrakis E, et al. Colistin serum concentrations after intravenous administration in critically ill patients with serious multidrug-resistant, gram-negative bacilli infections: a prospective, open-label, uncontrolled study. Clin Ther 2008;30:143–51.
76. Haas C, Kaufman D, Forrest A, et al. Colistin pharmaco-kinetics in the presence of renal and liver disease. Presented at the Society of Critical Care Medicine 39th critical care congress, Miami, FL, January 9–13, 2010.
77. Trotman R, Williamson J, Shoemaker M, et al. Antibiotic dosing in critically ill adult patients receiving continuous renal replacement therapy. Clin Infect Dis 2005;41:1159–66.
78. Li J, Milne RW, Nation RL, Turnidge JD, Smeaton TC, Coulthard K. Pharmacokinetics of colistin methanesulphonate and colistin in rats following an intravenous dose of colistin methanesulphonate. J Antimicrob Chemother 2004;53:837–40.
Posted: 12/09/2010; Pharmacotherapy. 2010;30(12):1279-1291. © 2010 Pharmacotherapy Publications
Abstract and Introduction
Abstract
Colistin is a polymyxin antibiotic that was discovered in the late 1940s for the treatment of gram-negative infections. After several years of clinical use, its popularity diminished because of reports of significant nephrotoxicity and neurotoxicity. Recently, the antibiotic has resurfaced as a last-line treatment option for multidrug-resistant organisms such as Pseudomonas aeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae. The need for antibiotics with coverage of these gram-negative pathogens is critical because of their high morbidity and mortality, making colistin a very important treatment option. Unfortunately, however, resistance to colistin has been documented among all three of these organisms in case reports. Although the exact mechanism causing colistin resistance has not been defined, it is hypothesized that the PmrA-PmrB and PhoP-PhoQ genetic regulatory systems may play a role. Colistin dosages must be optimized, as colistin is a last-line treatment option; in addition, suboptimal doses have been linked to the development of resistance. The lack of pharmacokinetic and pharmacodynamic studies and no universal harmonization of dose units, however, have made it difficult to derive optimal dosing regimens and specific dosing guidelines for colistin. In critically ill patients who may have multiorgan failure, renal insufficiency may alter colistin pharmacokinetics. Therefore, dosage alterations in this patient population are imperative to achieve maximal efficacy and minimal toxicity. With regard to colistin toxicity, most studies show that nephrotoxicity is reversible and less frequent than once thought, and neurotoxicity is rare. Further research is needed to fully understand the impact that the two regulatory systems have on resistance, as well as the dosages of colistin needed to inhibit and overcome these developing patterns.
Introduction
Antimicrobial resistance has become a worldwide health care crisis with many pathogens showing limited or no susceptibility to currently available antimicrobial treatments. Gram-negative infections are of even more concern because of the lack of effective treatments and the limited number of antibiotics in development to treat these potentially lethal pathogens.[1–5] No new antibiotics with activity against multidrug-resistant (MDR) gram-negative bacteria are expected to be released within the next 5 years. This emphasizes the need for last-line options, such as colistin, in cases where pathogens are resistant to all other antibiotics.
Colistin, a polymyxin antibiotic (polymyxin E), was first discovered in the 1940s but was not used clinically until the late 1950s. Historically, colistin was used to combat infections caused by problematic gram-negative bacteria. Reports of nephrotoxicity and neurotoxicity, however, deterred physicians from using the antibiotic, especially with the emergence of other antibiotics (e.g., aminoglycosides) that were less toxic. Between the 1970s and 1990s, colistin was not used often, and the number of studies analyzing its use and pharmacology was minimal.[5]
Recently, the lack of treatment options for MDR bacteria such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae, has led to the reemergence of colistin as an antimicrobial therapy. Because such a large gap exists between the years that colistin was used clinically, available pharmacokinetic and pharmacodynamic data are very limited. Thus, information regarding colistin toxicities and optimum dosing is not well defined, and no universal dosing for the antibiotic exists. In addition, reports have begun to surface of colistin resistance among the organisms that the drug is currently being used to treat.[1, 2] This increased rate of resistance has emphasized the need to provide adequate, effective dosing with minimal toxicity. To review the pharmacology, resistance, toxicities, pharmacodynamics, and dosing considerations associated with colistin, we performed a search of the MEDLINE database for journal articles published from 1945–May 2010.
Dosage Forms, Elimination, Mechanism of Action, and Spectrum of Activity
Colistin is available in two forms, colistin sulfate and colistimethate sodium, administered topically and parenterally, respectively. Both forms can be inhaled. It is extremely important to note that the two forms are not interchangeable. Colistin sulfate is cationic and stable, whereas colistimethate sodium is anionic and not stable in vitro or in vivo.[6, 7] Colistimethate sodium is the form that is safer to administer parenterally because of its lower rate of toxicity.[8] As a prodrug, colistimethate sodium is readily hydrolyzed to form partially sulfomethylated derivatives, as well as colistin sulfate, the active form of the drug.[8] This hydrolysis of colistimethate sodium to colistin is a very important step in providing the drug's antimicrobial activity. Until colistin is formed, colistimethate sodium by itself has been shown to display little to no antibacterial activity and is considered an inactive prodrug of colistin.[8]
Colistimethate sodium is eliminated mainly by the renal route, with a fraction of the dose being converted to active colistin in vivo. Colistin undergoes extensive renal tubular reabsorption and therefore is mainly cleared by nonrenal mechanisms.[9, 10] The mechanism behind colistin's bactericidal ability is thought to be indistinguishable from that of polymyxin B, the standard of the polymyxins.[11] Colistin is polycationic and has both hydrophilic and lipophilic moieties. These interact electrostatically with the outer membrane of gram-negative bacteria and competitively displace divalent cations from the membrane lipids, specifically calcium and magnesium.[12] This disrupts the outer membrane and releases lipopolysaccharides.[13] Change in the permeability of the bacterial membrane leads to leakage of the cell content and subsequently cell lysis and death.[2–4] Colistin also has the ability to bind and neutralize the lipopolysaccharide molecule of bacteria, giving it antiendotoxin activity.[2] Colistin has a narrow antibacterial spectrum of activity, with susceptibility mostly against common gram-negative isolates. Most significantly, it displays in vitro activity against MDR gram-negative pathogens such as A. baumannii, P.aeruginosa, and K. pneumoniae. Colistin also has activity against other isolates, such as Enterobacteriaceae,Stenotrophomonas maltophilia,Escherichia coli, Salmonella species, Shigella species, Haemophilus influenzae, Bordetellapertussis, and Legionella pneumophila.[2]
Reports of Colistin Resistance
As mentioned earlier, in the case of MDR gramnegative organisms such as A. baumannii, P.aeruginosa, and K. pneumoniae, the need for alternative treatments has led to the reemergence of colistin. Although colistin has been shown to be effective for the treatment of a wide variety of infections,[3, 14] its use for treating infections caused by these three gram-negative organisms has been impeded by occurrences of colistin resistance. Development of resistance to colistin is a serious concern. As colistin is the last line of defense against these virulent pathogens, resistance to this antibiotic may have devastating effects if no other treatment options are available to combat the infection. Cases of colistin resistance, as well as the mechanisms behind its development, are discussed in the following sections.
Acinetobacter baumannii
The increasing prevalence of A. baumannii infections coupled with its escalating resistance to available treatments and the lack of drug development to cover this pathogen has made it one of the most difficult gram-negative infections to treat and control.[1, 15] The Clinical and Laboratory Standards Institute susceptibility breakpoint for A. baumannii is 2 mg/L or lower, and the resistance breakpoint is 8 mg/L or higher.[16] Although colistin is often considered a reliable agent to treat A. baumannii, reports of resistant strains to this antibiotic are on the rise.[17–20] Recent studies have shown varying rates of resistance as well as the occurrence of hetero-resistant strains (Table 1).[15, 17–29]
In one study, 265 strains of Acinetobacter were collected from two Korean hospitals.[18] Of those 265 isolates, 214 (81%) were determined to be A.baumannii. With use of the broth microdilution method, 27.9% of these isolates were found to be resistant to colistin (minimum inhibitory concentration [MIC] > 16 mg/L) with a minimum concentration required to inhibit 90% of bacteria (MIC90) of 32 mg/L. The A. baumannii strains were further classified into three subgroups based on phylogenetic clustering. In subgroups II and III, most isolates were colistin resistant (64.8% and 88.9%, respectively) but typically remained susceptible to other conventional antibiotics (carbapenems, β-lactams, and ciprofloxacin). In contrast, the A. baumannii isolates in subgroup I had a much lower rate of resistance to colistin (7%), with an increased resistance profile to other antimicrobials.
Similar results were seen in a second study, which compared 17 colistin-resistant isolates of A. baumannii to 17 susceptible strains.[21] Like the previous study, this study also used the broth microdilution method for MICs. Generally, the strains resistant to colistin had increased susceptibility to conventional antibiotics, with substantial decreases in the MICs (up to 16 times lower). The colistin-resistant strains were also found to have a decreased ability to form biofilms, which is associated with diminished antibiotic susceptibility in A. baumannii.[21]
In several studies, colistin also exhibited heteroresistance.[15, 19, 22, 23] Heteroresistance occurs when subpopulations within the strain exhibit reduced susceptibility although the overall MIC is not altered. This makes detection of resistant subpopulations impossible with MIC alone. In one study of 16 A. baumannii isolates, all 16 strains were initially susceptible to colistin with an MIC of 2 mg/L or lower.[15] However, when concentrations of colistin up to 32 times MIC were used, significant regrowth of A.baumannii was noted at 24 hours. By the conclusion of the study, 15 of 16 isolates exhibited heteroresistance. Other studies had similar outcomes, with some heteroresistant strains showing regrowth within 6 hours, regardless of the dosing schedule initiated.[30–32]
A recent study of clinical isolates from the Western Pacific region showed 1 (3.3%) of 30 isolates to be resistant to colistin and 7 isolates (23%) to be colistin heteroresistant.[19] Although lower than that of previously reported cases of colistin resistance or heteroresistance, these findings still emphasize not only the need for adequate dosing, but also the potential use of combination therapy to eradicate these resistant subpopulations.
Pseudomonas aeruginosa
The high mortality rate associated with P.aeruginosa is in part related to its multiple mechanisms of resistance, with some clinical isolates showing panresistance to all United States Food and Drug Administration–approved antibiotics.[1] It has a Clinical and Laboratory Standards Institute susceptibility breakpoint of 2 mg/L or lower, and a resistance breakpoint of 4 mg/L or higher. Infections and resistance due to P. aeruginosa is of even more concern in patients with cystic fibrosis, as it is the most common colonizing pathogen in the lungs and has higher rates of resistance in this population.[25] Although colistin is usually regarded as salvage therapy and is sometimes the only therapeutic option to treat P. aeruginosa, cases of isolates resistant to colistin have emerged (Table 1).
In 385 strains of P. aeruginosa isolates from 57 adults with cystic fibrosis, only 34.9% of nonmucoid and 51.9% of mucoid strains were susceptible to colistin (MIC < 0.5 mg/L).[25] Furthermore, the MIC distribution pattern in this study showed two populations of MICs, which may be indicative of emerging resistance.
A second study of 23 clinical isolates from patients with cystic fibrosis found 11 of these strains to be resistant to colistin, with MICs exceeding 128 mg/L.[24] Also, cases of colistinresistant P. aeruginosa were seen in six children with cystic fibrosis after they received aerosolized colistin for a mean duration of 3.1 years.[26] This rise in colistin resistance by P. aeruginosa is beginning to surface in the cystic fibrosis population, possibly secondary to the widespread use of inhaled colistin in these patients. Because P. aeruginosa plays a large role in the lung destruction and eventual respiratory failure seen with cystic fibrosis, continued development of resistance would be detrimental.
Klebsiella pneumoniae
The need for alternative antimicrobials to treat K. pneumoniae has risen with the increased prevalence of K. pneumoniae carbapenemases-, extended-spectrum β-lactamase (ESBL)-, and metallo-β-lactamase (MBL)-producing strains of this bacteria. Although reports of colistin resistance with this pathogen are sparse, they are significant, as current and future treatment options for ESBL- and MBL-producing K.pneumoniae are limited.
In one study, 18 colistin-resistant (MIC > 8 mg/L) K. pneumoniae isolates were obtained from 13 patients over a 16-month period in an intensive care unit in Greece.[27] All of these patients had a long duration of both colistin treatment (median 27 days) and hospitalization (median 69 days), which likely contributed to the development of resistance. Most recently, 6.8% (15 of 221 isolates)[29] and 27.3% (6 of 22 isolates)[28] of collected K. pneumoniae isolates were found to be colistin resistant in South Korea and Australia, respectively.
Mechanisms of Resistance
Resistance to colistin can develop through adaptive or mutational mechanisms, with almost complete cross-resistance existing between colistin and other polymyxins.[4] A variety of gene mutations cause resistance to colistin by altering the outer membrane of gram-negative bacteria, which is colistin's site of action. Although data on the precise mechanism of resistance are scant and appear to be dependent on specific bacteria, the PmrA-PmrB and PhoP-PhoQ regulatory systems play important roles in its development (Table 2).[33–49] Two-component regulatory systems, such as PmrA-PmrB and PhoP-PhoQ, allow bacteria to respond to environmental changes by modifying the expression of genes. When these regulatory systems interact with one another, they have been shown to have even more profound effects.[33]
One mechanism of resistance involves changes in the structure of the bacteria's negatively charged surface lipopolysaccharides and lipid A. These modifications occur as a result of the activation of the PmrA-PmrB system, which is regulated by the PhoP-PhoQ system, but can also act independently in mildly acidic conditions or with high concentrations of iron.[34, 35] The PmrAPmrB system regulates two loci, PmrE and PmrHFIJKLM, which are responsible for the changes in lipid A and are essential for polymyxin resistance.[34, 36–40] When activated, the PmrA-PmrB system adds ethanolamine to the phosphate groups of the lipopolysaccharides and lipid A and also inserts aminoarabinose at the 4' phosphate of lipid A.[34, 35, 41, 42] These changes lower the overall charge of the lipopolysaccharide, thereby reducing the binding affinity of the cationic polymyxins.[43]
Environmental pH and magnesium concentrations are two environmental factors that appear to greatly affect the expression of the bacteria's genes and the subsequent development of resistance. In one study of Salmonella enterica grown in 10 mM magnesium chloride at a pH of 5.8, the organisms were approximately 100,000 times more resistant to polymyxin B than strains grown at a pH of 7.7.[41] This increase is attributed to increased activation of the PmrA-PmrB system at slightly acidic pH values and micromolar magnesium concentrations. It may be possible to monitor or correct pH and magnesium levels in order to help prevent resistance due to these environmental factors, but more information is needed before this can be determined. Because studies examining environmental pH and magnesium compared with the rate of resistance are limited, it remains unclear what steps clinicians should take. It is clear, however, that the effects of pH and magnesium require an active PmrA-PmrB system. The PmrA null mutants have failed to exhibit polymyxin resistance.[44]
Low magnesium concentrations also lead to the development of resistance by activating PhoP and PmrA, which not only modifies the bacteria's lipopolysaccharides but also increases the expression of a gene that has been shown to be a major factor in the development of resistance—the OprH gene.[33, 35, 37, 44] The OprH, PhoP, and PhoQ genes form an operon that is controlled by both PhoP and magnesium concentrations and contributes to polymyxin resistance.[35, 45] The OprH gene, which lies immediately downstream from the PhoP-PhoQ regulatory system, encodes an outer membrane protein, OprH, that has enhanced expression in low magnesium level conditions.[36] These OprH proteins occupy membrane magnesium sites and reduce the binding sites for colistin, therefore contributing to resistance.[35, 45–47]
The presence of exogenous polyamines (spermidine, spermine, putrescine, and cadaverine) has also been shown to induce the expression of the OprH-PhoP-PhoQ operon, resulting in increased MICs of not only polymyxins, but also aminoglycosides, quinolones, and fluorescent dyes against P.aeruginosa, regardless of the presence of cations.[48] Although OprH is presumed to play a role in resistance, it has been proven that its presence is not necessary for resistance to occur, since OprHdeficient strains of P. aeruginosa remain polymyxin resistant.[33] Similarly, studies have shown that although PhoP is essential for the transcription of the OprH-PhoP-PhoQ operon, PhoP-null strains of P. aeruginosa retain polymyxin resistance.[45] This is significant as it exemplifies the independent role that the PmrAPmrB system plays in polymyxin resistance, as well as the potential for other unidentified mechanisms of resistance.
Recently, the morphology and topography of colistin-resistant bacteria have been found to differ from that of colistin-susceptible cells, which could give us further insight into the genetic mechanisms leading to colistin resistance.[49] An atomic force microscopy study was performed of both colistin-resistant and colistin-susceptible strains at different growth phases.[49] Compared with spherically shaped colistin-resistant bacteria at early and mid-logarithmic phases, susceptible cells were found to be rod shaped with pili present at all phases. The number and length of pili for colistin-resistant cells were greatly reduced, which the authors note could be the reason colistinresistant cells are unable to form a biofilm. In addition, colistin-resistant cells had a greater topographic variability and finer surface texture. In the stationary phase, elongated worm-like cells were more prevalent in the susceptible group versus the resistant group, which showed more heterogeneity among the cells in this phase. Of interest, levels of bacterial outer membrane damage after treatment with colistin were similar for both susceptible and resistant cells, showing the ability of colistin-resistant cells to maintain interaction with the outer membrane.[21, 49] Based on these findings, it is evident that specific studies examining the genetic mechanisms behind these morphologic and topographic differences need to be performed, so that we may better understand the resistance associated with colistin.
Toxicity
Early use of colistin was linked to multiple reports of nephrotoxicity and neurotoxicity.[2] It was from this fear of toxicity that its use was halted shortly thereafter.[2–4, 50–52] Reports over the past decade, however, have shown that the toxicity associated with the polymyxins is much less than originally believed.[5, 50, 53–56] The results from earlier reports were most likely a result of a lack of pharmacokinetic, pharmacodynamic, and toxicity studies.[2] Also, incorrect dosing, which may have resulted directly from confusing dosage forms and units, and the presence of other nephrotoxic drugs or conditions may have contributed to these toxicities.[57] Several studies have since examined the safety of colistin, and their results give us further insight into the toxicities associated with the antibiotic (Table 3).[51, 53–56, 58–62]
Nephrotoxicity
Several studies have proven satisfactory safety profiles with intravenous colistimethate sodium 160 mg 3 times/day in patients with normal renal function.[58, 63, 64] The authors of two studies found that no serious adverse effects occurred with this dosage regimen in the cystic fibrosis populations that they studied, and that there were no notable changes in renal function.[58, 63] Furthermore, colistin has recently been found to have a more favorable toxicity profile compared with the aminoglycosides, which were originally used in place of colistin because of their suspected decrease in toxic effects.[55, 65] Another group found an observable decrease in renal function in patients receiving aminoglycosides, which was further worsened by coadministration of colistin.[55] Colistin used as monotherapy or in combination with nonnephrotoxic antibiotics, however, did not appear to cause renal damage. Two additional studies concluded that colistin was generally well tolerated in critically ill patients.[53, 59] In one of the studies,[59] serum creatinine level slightly increased by 0.25 mg/dl from baseline during treatment, but it is important to note that the study was performed in patients with decreased renal function.
Although this increase in creatinine level is of concern, no serious adverse effects occurred, and there were no data suggesting renal toxicity. Although the nephrotoxicity associated with colistin is not as toxic as originally thought, it is still an adverse effect that must be considered when administering the antibiotic. A few recent studies have given us more insight into colistin-induced renal impairment. One group of authors examined the occurrence of acute renal failure with use of the RIFLE—risk, injury, failure, loss, and end-stage kidney disease—criteria, by completing a retrospective review of patients (aged ≥ 18 yrs) who received intravenous colistimethate sodium (≥ 72 hrs) between January 2003 and December 2007.[60] Among the 66 patients, they found that the peak serum creatinine level during colistimethate sodium treatment met the RIFLE criteria for nephrotoxicity in 45% of patients, and that 21% of patients stopped colistimethate sodium therapy due to nephrotoxicity. The probability of renal toxicity increased in proportion to the overall total colistimethate sodium dose, and patients who received colistimethate sodium for longer than 14 days were 3.7 times more likely to experience nephrotoxicity. These findings are consistent with those of another group, who similarly found the change in serum creatinine level to be correlated with the cumulative dose of colistimethate sodium administered in their prospective cohort study.[61] Also, in the first study mentioned, the authors found that serum creatinine levels returned to baseline within 1 month after cessation of colistin,[60] suggesting reversibility of nephrotoxicity after discon- tinuation. A third group of authors performed a case-control study to evaluate the occurrence of nephrotoxicity and to analyze the characteristics and risk factors of patients who develop nephrotoxicity. [62] Hypoalbuminemia and concomitant use of a nonsteroidal antiinflammatory drug were the only statistically significant independent risk factors.
Neurotoxicity
Adverse effects such as paresthesias, visual alterations, ataxia, and neuromuscular blockade are possible with polymyxins as a class. These neurologic effects, however, are usually reversible after the cessation of treatment and usually occur in patients receiving prolonged treatment. Cases of neurotoxicity due to colistin have been mild and rare, with the current rate of neurotoxicity estimated to be 0–7%.[66] Little to no data exist on colistin alone causing neurotoxicity in patients. One study showed that in 21 patients treated with colistin for ventilator-associated pneumonia, there were no reports of neuromuscular blockade after patients were evaluated with an electro-physiologic study to detect the presence of neuromuscular transmission blockade and critical illness polyneuropathy.[67] Colistin was concluded to be a safe treatment alternative. Another study that included 17 patients who received colistin for more than 4 weeks of treatment, found that one patient appeared to develop neuropathy believed to be caused by colistin.[59] The authors concluded that despite this one finding, colistin was a safe and efficacious alternative therapy.
Summary
The toxicities associated with colistin have been found to be better correlated with the total cumulative amount of colistimethate sodium administered versus single or daily doses, and may occur more frequently in patients with hypoalbuminemia and concurrent nonsteroidal antiinflammatory drug use. Taking these risk factors into consideration when dosing can help to prevent adverse events. Although recent studies have found colistin-induced serum creatinine level increases, this adverse effect has been found to be reversible. Overall, many studies have shown that colistin is generally well tolerated, with less nephrotoxicity and neurotoxicity than was once thought.
Optimization of Colistin Dosing
Lack of Universal Dose Unit
We refer the reader to excellent review articles that extensively address this contemporary issue.[2, 5, 68] The limited data on colistin's pharmacokinetic and pharmacodynamic properties create immense confusion in assessing optimal dosing regimens that maximize antibacterial activity and minimize toxicity.[2, 5] Before attempting to determine optimal dosages for colistin, a universal dose unit measurement is needed when referring to the amount of drug being administered. The uncertainty related to dosing colistin is because some products use milligrams, whereas others use international units (IU). It has been established that there are approximately 12,500 IU per 1 mg of colistimethate sodium.[5] For example an average dose of colistimethate sodium is 2 million IU, which corresponds to 160 mg of drug.
To add further confusion, some products use milligrams of "colistin base activity" rather than milligrams of colistimethate sodium. It must be emphasized again that colistimethate sodium and colistin cannot be used interchangeably, especially when dosing. There are approximately 2.67 mg of colistimethate sodium per 1 milligram of colistin base.[5] To continue the example above, 2 million IU equals 160 mg of colistimethate sodium, which is equivalent to approximately 60 mg of colistin base. As can be seen by this complexity, the use of a unified dosage form and unit would greatly benefit the discussion of colistin dosing.
Discrepancies between Recommended Dosage Regimens
Once the proper dosage form and unit are established, the optimal dosage for patients must be decided. Because colistin is an older drug, there is little to no information on pharmacokinetics, pharmacodynamics, and toxicity to establish a safe and effective dosage regimen. Therefore, current dosage regimens are primarily derived from manufacturers' package inserts. The manufacturer of Colomycin (Xellia Pharmaceuticals, Copenhagen, Denmark) recommends that patients weighing more than 60 kg receive 1–2 million IU 3 times/day, equivalent to colistimethate sodium 80–160 mg 3 times/day, with a recommended daily upper limit of 6 million IU, or 480 mg of colistimethate sodium.[69] The manufacturer of Coly-Mycin M (Parkedale Pharmaceuticals, Inc., Rochester, MN) recommends 2.5–5 mg/kg/day colistin base activity in 2–4 divided doses, equivalent to colistimethate sodium 6.67–13.3 mg/kg/day or 83,375–166,250 IU/kg/day.[70] The recommended maximum daily dose of colistin is 10 million IU, or 800 mg of colistimethate sodium, from the manufacturer of Coly-Mycin M, which is approximately double the recommended daily dose from the manufacturer of Colomycin. Table 4 provides a comparison of the two products.
It should be of concern that the manufacturer of Coly-Mycin M recommends approximately double the dose of that recommended by the manufacturer of Colomycin. This lack of uniformity between manufacturers could lead to underdosing, inevitably leading to treatment failure and development of resistance.[68] More information is needed before we can begin to determine which regimens provide the best outcomes with acceptable safety.
Use of Pharmacodynamics to Guide Optimal Dosing
To fully optimize regimen selection of colistimethate sodium and colistin, it is important to appreciate the pharmacodynamics of colistin. Colistin is a rapidly bactericidal antimicrobial that possesses a significant postantibiotic effect against P. aeruginosa, A. baumannii, and K.pneumoniae. [24] The colistin area under the concentration-time curve (AUC):MIC ratio has been found to be the parameter best associated with efficacy.[71, 72] Researchers used neutropenic murine thigh and lung models to determine the pharmacokinetic-pharmacodynamic index of colistin that best correlates with efficacy against P.aeruginosa, and to determine the index target values needed for specific antibacterial effects.[71] For both the thigh and lung models, the unbound AUC:MIC (fAUC:MIC) ratio was the pharmacokinetic-pharmacodynamic index that had the strongest relationship to bacterial burden, with R2 values equaling 87% and 89% for the two models, respectively. The time where free drug concentration above the MIC (fT>MIC) was also closely correlated with efficacy, with R2 values equaling 84% and 88% for the thigh and lung models, respectively. The researchers noted, however, that fAUC:MIC ratio is still the pharmacokinetic-pharmacodynamic index most likely to be associated with colistin's activity because the scatter for the fT>MIC was relatively large in the 20–30% range, and because concentration-dependent killing has been seen with colistin in vitro. In the lung infection model, fAUC:MIC ratio target ranges for the three different strains of bacteria were 15.6–22.7, 27.6–36.1, and 53.3–66.7 for 1-log, 2-log, and 3- log bacterial kill, respectively. For the thigh infection model, target values ranged from 12.2–16.7, 36.9–45.9, and 105–141 for 1-log, 2-log, and 3-log kills, respectively. Although pharmacodynamic similarities among the two different sites of infection were seen, the observed differences emphasize the dosing alterations that may be needed based on site and type of infection.
Particularly in severe infections such as endocarditis, infections of prostheses, and ventilator-associated pneumonia, a high density of bacteria is known to exist, which may impact colistin pharmacodynamics.[73] Recently, the antibacterial activity of colistin was shown to be attenuated when facing a higher bacterial density. Investigators determined the extent and rate of killing by colistin to be greatly decreased at high compared with low inocula.[73] Against a genetically characterized clinical isolate of P. aeruginosa(PAO1), colistin killing was 23-fold slower at 109 and 6-fold slower at 108 compared with 106 colonyforming units, and 32-fold higher concentrations were required at 109 versus 106 colony-forming units. Although animal and in vivo studies are needed to further assess this inoculum effect, this study highlights the fact that higher colistin doses may be needed to treat sequestered, deep-seated infections with high bacterial densities.
When optimizing regimens for colistin, frequency of dosing is another important aspect to determine. One group of researchers evaluated the antibacterial activity and emergence of resistance that occurred with three different dosing intervals: 8, 12, and 24 hours.[72] Three different dosage regimens were used: 0.23 mg every 8 hours (0.30-mg loading dose), 0.39 mg every 12 hours (0.45-mg loading dose), and 0.89 mg every 24 hours (0.90-mg loading dose). The every-8-hour regimen simulated the expected maximum concentration ([Cmax] 3 mg/L) and minimum concentration (0.75 mg/L) at steady-state concentrations when colistin is given according to the manufacturer's recommendations. The every-12-hour and every-24-hour regimens were designed to provide higher target Cmax values (4.5 and 9.0 mg/L, respectively). The researchers found overall bacterial killing and regrowth to be similar among the three regimens. However, emergence of resistance increased as dosing interval increased, and the 8-hour regimen was the most effective at minimizing resistance. Concentrations remained above the MIC for approximately 80%, 72%, and 53% of the 72-hour treatment period for the 8-, 12-, and 24-hour dosage regimens, respectively. As colistin resistance continues to rise, these findings are important to keep in mind. In addition, other infection-specific considerations that can alter pharmacodynamics must be taken into account when dosing colistin and colistimethate sodium.
Dosing in Critically Ill Patients
In critically ill patients with multiorgan dysfunction and severe infections due to MDR organisms, treatment options are especially limited. Colistin remains a necessary last-line option for these patients. What is most concerning, however, is the lack of clinical guidelines and the presence of unclear dosing recommendations in this patient population.
Recent evidence has shown that the pharmacokinetics of colistimethate sodium and colistin in critically ill patients differ from those previously found among patients with cystic fibrosis.[68] In critically ill patients, who may have multiorgan failure, sepsis, or a wider range of renal impairment, the differences are important to take into account. Although the half-life of colistin is approximately 4 hours in patients with cystic fibrosis, it is longer in critically ill patients.[68] The half-life is 14.4 hours in critically ill patients, and the rate of formation of colistin from colistimethate sodium is different from previously published data.[74] In addition, larger volumes of distribution and lower concentrations of the antibiotic have been seen in critically ill patients with sepsis.[75] These differences have the ability to impact the effects of colistimethate sodium and colistin, which could require alterations in the dosage regimen. Although pharmacokinetic and pharmacodynamic data in this patient population are scarce, some studies have given us further insight into changes that may be needed when dosing critically ill patients.
One group of authors completed a population analysis to examine the pharmacokinetics of colistin after the administration of intravenous doses of colistimethate sodium in critically ill patients.[74] Patients received 3 million IU (240 mg) of colistimethate sodium intravenously every 8 hours or 160 mg every 8 hours if creatinine clearance was less than 50 ml/minute. The predicted plasma Cmax was 0.60 mg/L after the first dose and 2.3 mg/L at steady state. The authors found that after the first few doses of the regimen, colistin concentrations were below the Clinical and Laboratory Standards Institute MIC breakpoint of 2 mg/L for P. aeruginosa and Enterobacteriaceae. In addition, at steady state, plasma concentrations were below the MIC breakpoints for many of the cases. These results are of particular concern in critically ill patients, for whom a delay in appropriate treatment or suboptimal efficacy of the current regimen can lead to resistance and ultimately increased mortality. The authors speculated that a loading dose of colistimethate sodium is warranted. At 3 million IU every 8 hours, it would take 2–3 days before the steady-state concentration is achieved. Thus, the authors suggest that a colistimethate sodium loading dose of 9 or 12 million IU along with a 4.5 million IU maintenance dose every 12 concentration at a faster rate and with less frequent administration.
Similarly, another group assessed the steady-state serum concentrations of colistin after intravenous administration of colistimethate sodium 225 mg every 8 hours in 14 patients.[75] The average Cmax was found to be 2.93 mg/L, which the authors noted would most likely lead to suboptimal Cmax:MIC ratios for strains with higher MICs (e.g., A. baumannii and P.aeruginosa). The researchers concluded that higher doses of colistimethate sodium be considered. Based on these two studies, it is evident that further investigations using higher colistimethate sodium doses must be performed in critically ill patients to determine whether there is improved efficacy without increased toxicity.
The pharmacokinetic parameters of colistimethate sodium and colistin were examined in patients with stage 5 kidney disease or severe liver disease compared with healthy subjects.[76] Clearance of colistimethate sodium was found to be lower in the group of patients with kidney disease, and Cmax, half-life, and AUC were higher. In addition, conversion of colistimethate sodium to colistin and overall colistin exposure were increased in these patients, and clearance of colistin was decreased. Potentially, these results would have led to the neurotoxicity that occurred in the kidney disease group, as 3 of 10 patients in this group experienced paresthesias (which resolved in 24–48 hrs), compared with no patients in the liver disease group. Previously, a regimen of 2.5 mg/kg every 48 hours in patients receiving renal replacement therapy was suggested,[77] but this regimen has been found to be inadequate in some cases.[78] This study in particular highlights the fact that dosing may need to be altered in patients with renal failure.
In critically ill patients, for whom colistin's half-life appears to be longer, the potential for a longer dosing interval may be an option. Some studies, however, have found that as the interval between colistin doses becomes more extended, the prevalence of resistance increases.[72] This potentially serious consequence should be considered when deciding whether or not to use extended-interval dosing.
Overall, these data suggest that colistin pharmacokinetics are severely altered in critically ill patients. To maximize the AUC:MIC ratio, the predictive pharmacodynamic parameter of colistin, higher doses of colistimethate sodium and alterations in the dosing interval may be warranted. Because of colistin's toxicities, however, these may not be achievable. In these instances, combination therapy should be considered for optimal therapy and prevention of resistance.
Conclusion
Colistin has proved to be an important alternative for MDR gram-negative infections. However, reports of colistin-resistant strains have created a potentially dangerous scenario since it is the last line of defense. Colistin resistance is largely attributed to the PmrA-PmrB and PhoP-PhoQ regulatory systems and their responses to environmental changes. The activation of the PmrA-PmrB and PhoP-PhoQ regulatory systems produces resistance by activating a variety of genes that lower the negative charge of the outer membrane and decrease the number of binding sites for the cationic polymyxins. Although studies have begun to reveal the mechanisms behind colistin resistance, further research is needed to fully understand the impact that the two regulatory systems have on resistance, as well as the dosages of colistin needed to inhibit and overcome these developing patterns.
The development of colistin resistance has also been linked to inadequate dosing. This highlights the importance of dose optimization, especially in critically ill patients with MDR bacterial infections. Although higher doses appear beneficial, the lack of pharmacodynamic and pharmacokinetic data regarding colistin makes determination of appropriate dosing difficult. Colistin remains an essential alternative for most MDR gram-negative infections; however, cases of resistant strains should be a cause of much concern. Therefore, newer agents and colistin combination therapy are avenues that should be considered to optimize therapeutic regimens in the fight against evolving and highly resistant gram-negative infections.
References
1. Talbot GH, Bradley J, Edwards JE Jr, Gilbert D, Scheld M, Bartlett JG. Bad bugs need drugs: an update on the development pipeline from the antimicrobial availability task force of the Infectious Diseases Society of America. Clin Infect Dis 2006;42:657–68. (Erratum in Clin Infect Dis 2006;42: 1065.
2. Li J, Nation RL, Milne RW, Turnidge JD, Coulthard K. Evaluation of colistin as an agent against multi-resistant gram-negative bacteria. Int J Antimicrob Agents 2005;25:11–25.
3. Falagas ME, Kasiakou SK, Tsiodras S, Michalopoulos A. The use of intravenous and aerosolized polymyxins for the treatment of infections in critically ill patients: a review of the recent literature. Clin Med Res 2006;4:138–46.
4. Falagas ME, Kasiakou SK. Colistin: the revival of polymyxins for the management of multidrug-resistant gram-negative bacterial infections. Clin Infect Dis 2005;40:1333–41.
5. Li J, Nation RL, Turnidge JD, et al. Colistin: the re-emerging antibiotic for multidrug-resistant gram-negative bacterial infections. Lancet Infect Dis 2006;6:589–601.
6. Li J, Coulthard K, Milne R, et al. Steady-state pharmacokinetics of intravenous colistin methanesulphonate in patients with cystic fibrosis. J Antimicrob Chemother 2003;52:987–92.
7. Li J, Milne RW, Nation RL, Turnidge JD, Coulthard K. Stability of colistin and colistin methanesulfonate in aqueous media and plasma as determined by high-performance liquid chromatography. Antimicrob Agents Chemother 2003;47: 1364–70.
8. Bergen PJ, Li J, Rayner CR, Nation RL. Colistin methanesulfonate is an inactive prodrug of colistin against Pseudomonasaeruginosa. Antimicrob Agents Chemother 2006;50:1953–8.
9. Rodriguez CH, Pautaso J, Bombicino K, Vay C, Famiglietti A. Sensitivity to colistin: evaluation of cut-off points available in disk diffusion test [in Spanish]. Rev Argent Microbiol 2004;36:125–9.
10. Li J, Milne RW, Nation RL, Turnidge JD, Smeaton TC, Coulthard K. Use of high-performance liquid chromatography to study the pharmacokinetics of colistin sulfate in rats following intravenous administration. Antimicrob Agents Chemother 2003;47:1766–70.
11. Storm DR, Rosenthal KS, Swanson PE. Polymyxin and related peptide antibiotics. Annu Rev Biochem 1977;46:723–63.
12. Dixon RA, Chopra I. Leakage of periplasmic proteins from Escherichia coli mediated by polymyxin B nonapeptide. Antimicrob Agents Chemother 1986;29:781–8.
13. Peterson AA, Hancock RE, McGroarty EJ. Binding of polycationic antibiotics and polyamines to lipopolysaccharides of Pseudomonas aeruginosa. J Bacteriol 1985;164:1256–61.
14. Kasiakou SK, Rafailidis PI, Liaropoulos K, Falagas ME. Cure of post-traumatic recurrent multiresistant gram-negative rod meningitis with intraventricular colistin. J Infect 2005;50: 348–52.
15. Li J, Rayner CR, Nation RL, et al. Heteroresistance to colistin in multidrug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 2006;50:2946–50.
16. Galani I, Kontopidou F, Souli M, et al. Colistin susceptibility testing by Etest and disk diffusion methods. Int J Antimicrob Agents 2008;31:434–9.
17. Arroyo LA, Garcia-Curiel A, Pachon-Ibanez ME, et al. Reliability of the E-test method for detection of colistin resistance in clinical isolates of Acinetobacter baumannii. J Clin Microbiol 2005;43:903–5.
18. Ko KS, Suh JY, Kwon KT, et al. High rates of resistance to colistin and polymyxin B in subgroups of Acinetobacterbaumannii isolates from Korea. J Antimicrob Chemother 2007;60:1163–7.
19. Yau W, Owen RJ, Poudyal A, et al. Colistin heteroresistance in multidrug-resistant Acinetobacter baumannii clinical isolates from the western pacific region in the SENTRY antimicrobial surveillance programme. J Infect 2009;58:138–44.
20. Doi Y, Husain S, Potoski B, et al. Extensively drug-resistant Acinetobacter baumannii. Emerg Infect Dis 2009;15:980–2.
21. Li J, Nation RL, Owen RJ, Wong S, Spelman D, Franklin C. Antibiograms of multidrug-resistant clinical Acinetobacterbaumannii: promising therapeutic options for treatment of infection with colistin-resistant strains. Clin Infect Dis 2007;45:594–8.
22. Hawley JS, Murray CK, Griffith ME, et al. Susceptibility of Acinetobacter strains isolated from deployed U.S. military personnel. Antimicrob Agents Chemother 2007;51:376–8.
23. Hernan RC, Karina B, Gabriela G, Marcela N, Carlos V, Angela F. Selection of colistin-resistant Acinetobacter baumannii isolates in postneurosurgical meningitis in an intensive care unit with high presence of heteroresistance to colistin. Diagn Microbiol Infect Dis 2009;65:188–91.
24. Li J, Turnidge J, Milne R, Nation RL, Coulthard K. In vitro pharmacodynamic properties of colistin and colistin methanesulfonate against Pseudomonas aeruginosa isolates from patients with cystic fibrosis. Antimicrob Agents Chemother 2001;45:781–5.
25. Schulin T. In vitro activity of the aerosolized agents colistin and tobramycin and five intravenous agents against Pseudomonasaeruginosa isolated from cystic fibrosis patients in southwestern Germany. J Antimicrob Chemother 2002;49:403–6.
26. Denton M, Kerr K, Mooney L, et al. Transmission of colistin-resistant Pseudomonas aeruginosa between patients attending a pediatric cystic fibrosis center. Pediatr Pulmonol 2002;34: 257–61.
27. Antoniadou A, Kontopidou F, Poulakou G, et al. Colistin-resistant isolates of Klebsiella pneumoniae emerging in intensive care unit patients: first report of a multiclonal cluster. J Antimicrob Chemother 2007;59:786–90.
28. Poudyal A, Howden B, Bell J, et al. In vitro pharmacodynamics of colistin against multidrug-resistant Klebsiella pneumoniae. J Antimicrob Chemother 2008;62:1311–18.
29. Suh J-Y, Son J, Chung D, et al. Nonclonal emergence of colistin-resistant Klebsiella pneumoniae isolates from blood samples in South Korea. Antimicrob Agents Chemother 2010;54:560–2.
30. Owen RJ, Li J, Nation RL, Spelman D. In vitro pharmaco-dynamics of colistin against Acinetobacter baumannii clinical isolates. J Antimicrob Chemother 2007;59:473–7.
31. Tan TY, Ng LS, Tan E, Huang G. In vitro effect of minocycline and colistin combinations on imipenem-resistant Acinetobacterbaumannii clinical isolates. J Antimicrob Chemother 2007;60:421–3.
32. Tan CH, Li J, Nation RL. Activity of colistin against heteroresistant Acinetobacter baumannii and emergence of resistance in an in vitro pharmacokinetic/pharmacodynamic model. Antimicrob Agents Chemother 2007;51:3413–15.
33. Macfarlane EL, Kwasnicka A, Ochs MM, Hancock RE. PhoP-PhoQ homologues in Pseudomonas aeruginosa regulate expression of the outer-membrane protein OprH and polymyxin B resistance. Mol Microbiol 1999;34:305–16.
34. Gunn JS, Ryan SS, Van Velkinburgh JC, Ernst RK, Miller SI. Genetic and functional analysis of a PmrA-PmrB-regulated locus necessary for lipopolysaccharide modification, antimicrobial peptide resistance, and oral virulence of Salmonella enterica Serovar Typhimurium. Infect Immun 2000;68:6139–46.
35. Soncini FC, Groisman EA. Two-component regulatory systems can interact to process multiple environmental signals. J Bacteriol 1996;27:6796–801.
36. McPhee JB, Bains M, Winsor G, et al. Contribution of the PhoP-PhoQ and PmrA-PmrB two-component regulatory systems to Mg2+-induced gene regulation in Pseudomonasaeruginosa. J Bacteriol 2006;188:3995–4006.
37. McPhee JB, Lewenza S, Hancock RE. Cationic antimicrobial peptides activate a two-component regulatory system, PmrAPmrB, that regulates resistance to polymyxin B and cationic antimicrobial peptides in Pseudomonas aeruginosa. Mol Microbiol 2003;50:205–17.
38. Moskowitz SM, Ernst RK, Miller SI. PmrAB, a two-component regulatory system of Pseudomonas aeruginosa that modulates resistance to cationic antimicrobial peptides and addition of aminoarabinose to lipid A. J Bacteriol 2004;186:575–9.
39. Trent MS, Ribeiro AA, Lin S, Cotter RJ, Raetz CR. An inner membrane enzyme in Salmonella and Escherichia coli that transfers 4-amino-4-deoxy-L-arabinose to lipid A: induction on polymyxin-resistant mutants and role of a novel lipid-linked donor. J Biol Chem 2001;276:43122–31.
40. Zhang L, Dhillon P, Yan H, Farmer S, Hancock RE. Interactions of bacterial cationic peptide antibiotics with outer and cytoplasmic membranes of Pseudomonas aeruginosa. Antimicrob Agents Chemother 2000;44:3317–21.
41. Perez JC, Groisman EA. Acid pH activation of the PmrA/PmrB two-component regulatory system of Salmonella enterica. Mol Microbiol 2007;63:283–93.
42. Gunn JS, Miller SI. PhoP-PhoQ activates transcription of pmrAB, encoding a two-component regulatory system involved in Salmonella typhimurium antimicrobial peptide resistance. J Bacteriol 1996;178:6857–64.
43. Gunn JS, Lim KB, Krueger J, et al. PmrA-PmrB-regulated genes necessary for 4-aminoarabinose lipid A modification and polymyxin resistance. Mol Microbiol 1998;27:1171–82.
44. Groisman EA, Kayser J, Soncini FC. Regulation of polymyxin resistance and adaptation to low-Mg2+ environments. J Bacteriol 1997;179:7040–5.
45. Macfarlane EL, Kwasnicka A, Hancock RE. Role of Pseudomonas aeruginosa PhoP-phoQ in resistance to antimicrobial cationic peptides and aminoglycosides. Microbiology 2000;146(pt 10):2543–54.
46. Nicas TI, Hancock RE. Alteration of susceptibility to EDTA, polymyxin B and gentamicin in Pseudomonas aeruginosa by divalent cation regulation of outer membrane protein H1. J Gen Microbiol 1983;129:509–17.
47. Nicas TI, Hancock RE. Outer membrane protein H1 of Pseudomonas aeruginosa: involvement in adaptive and mutational resistance to ethylenediaminetetraacetate, polymyxin B, and gentamicin. J Bacteriol 1980;143:872–8.
48. Kwon DH, Lu CD. Polyamines induce resistance to cationic peptide, aminoglycoside, and quinolone antibiotics in Pseudomonas aeruginosa PAO1. Antimicrob Agents Chemother 2006;50:1615–22.
49. Soon RL, Nation RL, Hartley PG, Larson I, Li J. Atomic force microscopy investigation of the morphology and topography of colistin-heteroresistant Acinetobacter baumannii strains as a function of growth phase and in response to colistin treatment. Antimicrob Agents Chemother 2009;53:4979–86.
50. Falagas ME, Kasiakou SK. Toxicity of polymyxins: a systematic review of the evidence from old and recent studies [online exclusive article]. Crit Care 2006;10:R27. Available from http://ccforum.com/content/10/1/R27.
51. Kasiakou SK, Michalopoulos A, Soteriades ES, Samonis G, Sermaides GJ, Falagas ME. Combination therapy with intravenous colistin for management of infections due to multidrug-resistant gram-negative bacteria in patients without cystic fibrosis. Antimicrob Agents Chemother 2005;49: 3136–46.
52. Falagas ME, Bliziotis IA, Kasiakou SK, Samonis G, Athanassopoulou P, Michalopoulos A. Outcome of infections due to pandrug-resistant (PDR) gram-negative bacteria [online exclusive article]. BMC Infect Dis 2005;5:24. Available from http://www.biomedcentral.com/1471-2334/5/24.
53. Reina R, Estenssoro E, Saenz G, et al. Safety and efficacy of colistin in Acinetobacter and Pseudomonas infections: a prospective cohort study. Intensive Care Med 2005;31:1058–65.
54. Cheng C-Y, Sheng W-H, Wang J-T, et al. Safety and efficacy of intravenous colistin (colistin methanesulphonate) for severe multidrug-resistant gram-negative bacterial infections. Int J Antimicrob Agents 2010;25:297–300.
55. Al-Aloul M, Miller H, Alapati S, Stockton PA, Ledson MJ, Walshaw MJ. Renal impairment in cystic fibrosis patients due to repeated intravenous aminoglycoside use. Pediatr Pulmonol 2005;39:15–20.
56. Santamaria C, Mykietiuk A, Temporiti E. Nephrotoxicity associated with the use of intravenous colistin. Scand J Infect Dis 2009;41:767–9.
57. Li J, Rayner CR, Nation RL. Colistin-associated acute renal failure: revisited. South Med J 2005;98:1229–30.
58. Conway SP, Etherington C, Munday J, Goldman MH, Strong JJ, Wootton M. Safety and tolerability of bolus intravenous colistin in acute respiratory exacerbations in adults with cystic fibrosis. Ann Pharmacother 2000;34:1238–42.
59. Falagas ME, Rizos M, Bliziotis IA, Rellos K, Kasiakou SK, Michalopoulos A. Toxicity after prolonged (more than four weeks) administration of intravenous colistin [online exclusive article]. BMC Infect Dis 2005;5:1. Available from http://www.biomedcentral.com/1471-2334/5/1.
60. Hartzell J, Neff R, Ake J. Nephrotoxicity associated with intravenous colistin (colistimethate sodium) treatment at a tertiary care medical center. Clin Infect Dis 2009;48:1724–8.
61. Falgas M, Fragoulis K, Kasiakou S, et al. Nephrotoxicity of intravenous colistin: a prospective evaluation. Int J Antimicrob Agents 2005;26:504–7.
62. Kim J, Lee K-H, Yoo S, et al. Clinical characteristics and risk factors of colistin-induced nephrotoxicity. Int J Antimicrob Agents 2009;34:434–8.
63. Ledson MJ, Gallagher MJ, Cowperthwaite C, Convery RP, Walshaw MJ. Four years' experience of intravenous Colomycin in an adult cystic fibrosis unit. Eur Respir J 1998;12:592–4.
64. Conway SP, Pond MN, Watson A, Etherington C, Robey HL, Goldman MH. Intravenous colistin sulphomethate in acute respiratory exacerbations in adult patients with cystic fibrosis. Thorax 1997;52:987–93.
65. Assadamongkol K, Tapaneya-Olarn W, Chatasingh S. Urinary N-acetyl-beta-D-glucosaminidase (NAG) in aminoglycoside nephrotoxicity. J Med Assoc Thai 1989;72(suppl 1):42–6.
66. Molina J, Cordero E, Pachon J. New Information about the polymyxin/colistin class of antibiotics. Expert Opin Pharmacother 2009;10:2811–28.
67. Garnacho-Montero J, Ortiz-Leyba C, Jimenez-Jimenez FJ, et al. Treatment of multidrug-resistant Acinetobacter baumannii ventilator-associated pneumonia (VAP) with intravenous colistin: a comparison with imipenem-susceptible VAP. Clin Infect Dis 2003;36:1111–18.
68. Nation R, Li J. Colistin in the 21st century. Curr Opin Infect Dis 2009 2009;22:535–43.
69. Forest Laboratories Ltd. Colomycin (colistimethate sodium) injection summary of product characteristics. Bexley, Kent, United Kingdom; 2010.
70. Monarch Pharmaceuticals. Coly-Mycin M (colistimethate) parenteral package insert. Bristol, TN; 2005.
71. Dudhani RV, Turnidge JD, Coulthard K, et al. Elucidation of the pharmacokinetic/pharmacodynamic determinant of colistin activity against Pseudomonas aeruginosa in murine thigh and lung infection models. Antimicrob Agents Chemother 2010;54:1117–24.
72. Bergen PJ, Li J, Nation RL, Turnidge JD, Coulthard K, Milne RW. Comparison of once-, twice- and thrice-daily dosing of colistin on antibacterial effect and emergence of resistance: studies with Pseudomonas aeruginosa in an in vitro pharmaco-dynamic model. J Antimicrob Chemother 2008;61:636–42.
73. Bulitta JB, Yang JC, Yohonn L, et al. Attenuation of colistin bactericidal activity by high inoculum of Pseudomonasaeruginosa characterized by a new mechanism-based population pharmacodynamic model. Antimicrob Agents Chemother 2010;54:2051–62.
74. Plachouras D, Karvanen M, Friberg L, et al. Population pharmacokinetic analysis of colistin methanesulfonate and colistin after intravenous administration in critically ill patients with infections caused by gram-negative bacteria. Antimicrob Agents Chemother 2009;53:3430–6.
75. Markou N, Markantonis S, Dimitrakis E, et al. Colistin serum concentrations after intravenous administration in critically ill patients with serious multidrug-resistant, gram-negative bacilli infections: a prospective, open-label, uncontrolled study. Clin Ther 2008;30:143–51.
76. Haas C, Kaufman D, Forrest A, et al. Colistin pharmaco-kinetics in the presence of renal and liver disease. Presented at the Society of Critical Care Medicine 39th critical care congress, Miami, FL, January 9–13, 2010.
77. Trotman R, Williamson J, Shoemaker M, et al. Antibiotic dosing in critically ill adult patients receiving continuous renal replacement therapy. Clin Infect Dis 2005;41:1159–66.
78. Li J, Milne RW, Nation RL, Turnidge JD, Smeaton TC, Coulthard K. Pharmacokinetics of colistin methanesulphonate and colistin in rats following an intravenous dose of colistin methanesulphonate. J Antimicrob Chemother 2004;53:837–40.
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