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* Bacteria growing less susceptible to common antiseptic

Chlorhexidine gluconate (CHG) has been increasingly used in hospitals in light of recent evidence that daily antiseptic baths for patients in intensive care units (ICUs) may prevent infections and stop the spread of healthcare-associated infections. The impact of this expanded use on the effectiveness of the disinfectant is not yet known. “Hospitals are appropriately using chlorhexidine to reduce infections and control the spread of antibiotic-resistant organisms,” said Nuntra Suwantarat, MD, lead author. “However, our findings are a clear signal that we must continue to monitor bacteria for emerging antiseptic resistance as these antibacterial washes become more widely used in hospitals.” In the study, investigators compared bacterial resistance between cultures from patients in eight ICUs receiving daily antiseptic washes to patients in 30 non-ICUs who did not bathe daily with CHG. Bacterial cultures obtained from patients with regular antiseptic baths showed reduced susceptibility to CHG when compared with those from patients who did not have antiseptic baths. Regardless of unit protocol, 69 percent of all bacteria showed reduced CHG susceptibility, a trend that requires vigilant monitoring. “The good news is that most bacteria remain vulnerable to CHG, despite the reduced susceptibility. Daily baths with a CHG solution remain effective against life-threatening bloodstream infections,” said Suwantarat. The investigators caution that the clinical implications of their findings remain unclear. For example, antibiotic susceptibility tests are commonly used to determine whether patients will respond to antibiotic treatment. A similar correlation between antiseptic susceptibility and response to an antiseptic are not as well defined. Identifying particular bacteria and settings in which these bacteria will not respond to antiseptic agents used in hospitals is an important next step.

http://www.sciencedaily.com/  Science Daily

http://www.sciencedaily.com/releases/2014/08/140813103940.htm  Original web page at Science Daily

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Harnessing the power of bacteria’s sophisticated immune system

Bacteria’s ability to destroy viruses has long puzzled scientists, but researchers at the Johns Hopkins Bloomberg School of Public Health say they now have a clear picture of the bacterial immune system and say its unique shape is likely why bacteria can so quickly recognize and destroy their assailants. The researchers drew what they say is the first-ever picture of the molecular machinery, known as Cascade, which stands guard inside bacterial cells. To their surprise, they found it contains a two-strand, unencumbered structure that resembles a ladder, freeing it to do its work faster than a standard double-helix would allow. The findings, published online Aug. 14 in the journal Science, may also provide clues about the spread of antibiotic resistance, which occurs when bacteria adapt to the point where antibiotics no longer work in people who need them to treat infections, since similar processes are in play. The World Health Organization (WHO) considers antibiotic resistance a major threat to public health around the world. “If you understand what something looks like, you can figure out what it does,” says study leader Scott Bailey, PhD, an associate professor in the BloombergSchool’s Department of Biochemistry and Molecular Biology. “And here we found a structure that nobody’s ever seen before, a structure that could explain why Cascade is so good at what it does.” For their study, Bailey and his colleagues used something called X-ray crystallography to draw the picture of Cascade, a key component of bacteria’s sophisticated immune system known as CRISPR, an acronym for Clustered Regularly Interspaced Short Palindromic Repeats. Cascade uses the information housed in sequences of RNA as shorthand to identify foreign invaders and kill them. Much of the human immune system is well understood, but until recently scientists didn’t realize the level of complexity associated with the immune system of single-cell life forms, including bacteria. Scientists first identified CRISPR several years ago when trying to understand why bacterial cultures used to make yogurt succumbed to viral infections. Researchers subsequently discovered they could harness the CRISPR bacterial immune system to edit DNA and repair damaged genes. One group, for example, was able to remove viral DNA from human cells infected with HIV. Bailey’s work is focused on how Cascade is able to help bacteria fight off viruses called bacteriophages. The Cascade system uses short strands of bacterial RNA to scan the bacteriophage DNA to see if it is foreign or self. If foreign, the cell launches an attack that chews up the invading bacteriophage.

To “see” how this happens, Bailey and his team converted Cascade into a crystalized form. Technicians at the National Synchrotron Light Source at Brookhaven National Laboratory in Upton, NY, and the Stanford Synchrotron Radiation Lightsource then trained high-powered X-rays on the crystals. The X-rays provided computational data to the BloombergSchool scientists allowing them to draw Cascade, an 11-protein machine that only operates if each part is in perfect working order. What they saw was unexpected. Instead of the RNA and DNA wrapping around each other to form what is known as a double-helix structure, in Cascade the DNA and RNA are more like parallel lines, forming something of a ladder. Bailey says that if RNA had to wrap itself around DNA to recognize an invader — and then unwrap itself to look at the next strand — the process would take too much time to ward off infection. With a ladder structure, RNA can quickly scan DNA. In the new study, Bailey says his team determined that the RNA scans the DNA in a manner similar to how humans scan text for a key word. They break long stretches of characters into smaller bite-sized segments, much like words themselves, so they can be spotted more easily. Since the CRISPR-Cas system naturally acts as a barrier to the exchange of genetic information between bacteria and bacteriophages, its function can offer clues to how antibiotic resistance develops and ideas for how to keep it from happening. “We’re finding more pieces to the puzzle,” Bailey says. “This gives us a better understanding of how these machines find their targets, which may help us harness the CRISPR system as a tool for therapy or manipulation of DNA in a lab setting. And it all started when someone wanted to make yogurt more cheaply.”

http://www.sciencedaily.com/  Science Daily

http://www.sciencedaily.com/releases/2014/08/140814191350.htm  Original web page at Science Daily

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* Highly drug-resistant, virulent strain of Pseudomonas aeruginosa arises in Ohio

A team of clinician researchers has discovered a highly virulent, multidrug resistant form of the pathogen, Pseudomonas aeruginosa, in patient samples in Ohio. Their investigation suggests that the particular genetic element involved, which is still rare in the United   States, has been spreading heretofore unnoticed, and that surveillance is urgently needed. The research is published ahead of print in Antimicrobial Agents and Chemotherapy. The P. aeruginosa contained a gene for a drug resistant enzyme called a metallo beta-lactamase. Beta-lactamases enable broad-spectrum resistance to beta-lactam antibiotics, including carbapenems, cephalosporins, and penicillins, because they can break the four atom beta-lactam ring, a critical component of these antibiotics’ structure. The initial isolate of metallo-beta-lactamase-producing P. aeruginosa was identified in March, 2012, in a foot wound of a 69-year-old man with type 2 diabetes living in a long-term care facility. During 2012-2013, the investigators identified this highly antibiotic-resistant infection in six other patients. One of the seven patients subsequently died of the infection. The cases are linked epidemiologically via admission to a community hospital and residence in long-term care facilities in Northeast Ohio. The one exception was a patient from Qatar who was transferred into a tertiary medical center in Ohio, says lead author Federico Perez, of the Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Cleveland, Ohio, USA. The investigators subsequently found that the metallo beta-lactamase was contained within an integron, a genetic element that can jump from one species of bacterium to another, can reside on plasmids or within the chromosomes, and is known for being able to contain multiple antibiotic resistance genes. This particular metallo beta-lactamase, verona integron-encoded metallo beta-lactamse (VIM), is widespread globally, if not in the US. “VIM enzymes confer resistance to imipenem and all other beta-lactams,” says Perez. “They are not inhibited by metallo beta-lactamase inhibitors.” “Alarmingly, the extensively drug-resistant phenotype expressed by some of these isolates precluded any reliable antibiotic treatment since they even displayed intermediate resistance to colistin, an ‘agent of last resort’,” the researchers write. “Patients who were affected had multiple comorbidities, endured prolonged colonization, required long-term care and, in one instance had a lethal outcome from a bloodstream infection.”

On top of everything else, genomic sequencing and assembly showed that the integron was part of a novel 35 kilobase region that included a transposon (another mobile genetic element) and the so-called Salmonella Genomic Island 2 (SGI2). That indicated that a recombination event had occurred between Salmonella and P. aeruginosa, contributing even more resistance genes to the latter. “This is the first description of genetic exchange of a large mobile element — the Salmonella Genome Island — and resistance genes between P. aerugenosa and Salmonella, says Perez. “This movement of genetic material creates concern that metallo beta-lactamases will disseminate rapidly in these enteric pathogens that are also very invasive. We are also concerned about the possibility of enhanced virulence.”

http://www.sciencedaily.com/  Science Daily

http://www.sciencedaily.com/releases/2014/08/140811170209.htm  Original web page at Science Daily

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* Contamination hits cell work

Mycoplasma infestations are widespread and costing laboratories millions of dollars in lost research. John Hogenesch saw the anguished look on his technician’s face and knew instantly that her experiments had gone haywire; he also had a pretty good idea of why. “Check your culture for Mycoplasma contamination,” he advised. The bacterium is notorious for infecting cell cultures, and had indeed compromised her experiments. In fact, the problem is widespread. Hogenesch, a genome biologist at the University of Pennsylvania in Philadelphia and his colleague Anthony Olarerin-George have found that more than one-tenth of gene-expression studies, many published in leading journals, show evidence of Mycoplasma contamination. The infestations are undermining research findings and wasting huge amounts of money, Hogenesch says. He should know. His lab quickly overcame an infestation last year, but a previous plague cost it some US$100,000 and a year of research. Mycoplasma takes hold quickly, he says. “All it takes is one person not to check, and — bam — you have it.” The bacterium often comes from lab workers, and is not killed by the antibiotics typically used to rid cell cultures of contaminants. And unlike many other microorganisms, which turn the growth medium turbid, Mycoplasma leaves no visible signs of its presence. Mycoplasma is a long-standing problem. The bacterium plagued early cultures of HeLa cells, a widely used human cell line established in the 1950s. Surveys of individual collections have long served as a warning: a 1993 study, for instance, found Mycoplasma in 15% of 20,000 cultures from US Food and Drug Administration labs. Companies and centres that distribute cells and reagents are now fastidious in screening for it. To get a global perspective on the problem, Hogenesch and Olarerin-George looked for stretches of Mycoplasma DNA in RNA-sequence data from more than 9,000 samples — collected during experiments done between 2012 and 2013 to measure gene expression in cultured mammalian cells. A total of 11% of the samples were found to contain Mycoplasma DNA at levels indicative of contamination.

Some of the studies with the highest levels were published in leading journals such as Cell, Nature and Proceedings of the National Academy of Sciences, says Hogenesch. Contamination does not necessarily invalidate those findings, but the bacterium can influence the expression of hundreds of genes and hinders cell growth by competing for nutrients. In one particularly contaminated data set, of individual lymphoma cells, Hogenesh and Olarerin-George identified 61 genes whose levels were altered by Mycoplasma. Their work is available on the bioRχiv preprint server, and they plan to submit their findings to a peer-reviewed journal soon. If more than one-tenth of cell cultures are contaminated, the costs in wasted time and resources, such as repeating experiments and replacing cells, could run into hundreds of millions of dollars, says Hogenesch. In 2013, for example, the US National Institutes of Health spent about $3 billion on research that uses cell lines, and Hogenesch estimates that about one-third of his lab costs go on tissue culture. Contamination is a result of “sloppy cell-culture work”, says Hans Drexler, a physician–scientist at the German Collection of Microorganisms and Cell Cultures in Braunschweig. “Mycoplasma don’t fall from the sky,” he says. “They are introduced into the cell culture by people.” He also is not surprised that so many cell cultures are contaminated. A similar percentage of the human leukaemia and lymphoma cell lines his lab received from other researchers between 2010 and 2013 tested positive for Mycoplasma, he says. But this is an improvement: one-quarter of such cultures were tainted in the early 1990s, he found. Drexler believes that Mycoplasma contamination persists because of a “black market” in cell lines — researchers often share cultures in violation of materials-transfer agreements. He estimates that 10% of the cell lines he receives from these sources are contaminated, compared with none from official suppliers. To solve the problem, he urges labs to spend the extra money on cell lines from reputable sources, and test those they have for contamination. “There’s no magic twenty-first-century bullet that’s going to kill these things,” Hogenesch says. “We have to be continuously vigilant, clean up the cultures that have them, and destroy the bacteria altogether.”

Nature 511, 518 (31 July 2014) doi:10.1038/511518a

http://www.nature.com/news/index.html  Nature

http://www.nature.com/news/contamination-hits-cell-work-1.15632  Original web page at Nature

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C. difficile vaccine proves safe, 100 percent effective in animal models

An experimental vaccine protected 100 percent of animal models against the highly infectious and virulent bacterium, Clostridium difficile, which causes an intestinal disease that kills approximately 30,000 Americans annually. The research is published ahead of print in Infection and Immunity. In the study, the vaccine protected the mice and non-human primates against the purified toxins produced by C. difficile, as well as from an orogastric spore infection, a laboratory model that mimics the human disease, after only two immunizations. “Animals that received two immunizations did not get sick or show signs of C. difficile-associated disease,” says corresponding author Michele Kutzler, of Drexel University College of Medicine, Philadelphia. “While our research was conducted in animal models, the results are very translatable to the clinic,” says Kutzler. “In some cases, patients who acquire C. difficile can develop serious complications including severe diarrhea, toxic megacolon, bowel perforation, multi-organ failure, and death. Once fully developed, our DNA vaccine could prevent the deadly effects of C. difficile infection when administered to hospital patients at risk of acquiring C. difficile.” The protection following just two immunizations is especially important since the time window in humans between colonization with C. difficile and the onset of disease symptoms can be a mere 10-14 days, says Kutzler. The vaccine protects against the bacterial toxins by mustering anti-toxin neutralizing antibodies, says Kutzler. The cost of fighting the half million C. difficile infections that occur annually in the US is estimated to be nearly $10 billion, most of which could be saved by a successful preventive vaccine, says Kutzler. Morbidity and mortality have risen over the last decade, likely due to increased prevalence of relapsing disease, and hypervirulent strains, she adds. Treating the disease is especially difficult, as the bacterial spores persist in the hospital environment, where most infections occur. There is no standard, effective treatment for recurrent disease, but a small number of experimental fecal transplants for C. difficile have had a very high success rate, with no adverse reactions. “Since our vaccine was safe, effective after only two immunizations, and performed exceptionally well, we feel that this success warrants further studies using human patients,” says Kutzler.

http://www.sciencedaily.com/  Science Daily

http://www.sciencedaily.com/releases/2014/07/140731145933.htm  Original web page at Science Daily

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A cure for the plague of frogs?

One of the worst scourges of frogs and their kin is Batrachochytrium dendrobatidis (Bd), a deadly fungus that infects nearly half of amphibian species, eats away their skin, and causes heart attacks. Now, a study shows that one kind of frog can learn to avoid the widespread fungus and that two species become resistant with repeated exposures. Although preliminary, the findings suggest that there may be a way to help protect more vulnerable amphibians. “If frogs exposed and cured are therefore ‘vaccinated’ against Bd, then this study has big implications for mitigation of Bd outbreaks in the future and restoration of frog populations that are in captive holding now,” writes biologist Kelly Zamudio of Cornell University, who was not involved in the study, in an e-mail. Amphibians suffer from many threats, including pollution and habitat destruction. It isn’t easy to pin an extinction on Bd, but researchers suspect the fungus is behind the mysterious decline of many species. To study how well frogs can resist the fungus, Jason Rohr, an ecologist at the University of South Florida, Tampa, and colleagues set up experiments with two species that are easy to catch in Florida, the oak toad (Bufo quercicus) and the Cuban tree frog (Osteopilus septentrionalis). First, they investigated whether oak toads could learn to avoid the fungus. After setting up a small chamber in their laboratory, they added the fungus to one side. When oak toads were first put into the chamber, they spent equal amount of time on either side. Then, Rohr and his colleagues removed the toads and killed the fungus with moderate heat. When the same toads were placed back into the chamber, they were less likely to spend time on the side with the fungus, suggesting that they can learn to avoid it. Rohr has additional experiments under way to determine how the toads detect the fungus, but he suspects that contact is painful as the fungus attacks. “It can’t feel good to have your skin digested by an enzyme.” Other trials with both species showed that with each exposure to the fungus, the toad’s and frog’s immune systems strengthened their response. The animals roughly doubled the number of immune cells by the fourth time they encountered the fungus. And a greater proportion of the amphibians survived: Although only 20% made it through the first exposure, more than half escaped their fourth bout with the fungus, the team reports in this week’s issue of Nature.

This finding isn’t relevant to oak toads and Cuban tree frogs in the wild, because the weather is hot enough where they live to usually kill the fungus. But it could help explain how some species in colder environments, such as mountain lakes, have managed to survive. An intriguing finding is that exposure to dead Bd can also increase resistance, which raises the prospect of immunizing highly sensitive species. “That is exciting stuff,” says biologist Karen Lips of the University of Maryland, College Park, although she describes the level of protection as moderate. Rohr hopes to try adding large amounts of dead Bd to ponds to test whether it will help the survival of susceptible frogs. Several important questions remain. It’s not clear whether the resistance is permanent, whether tadpoles can become resistant, or whether it occurs in other species and would be strong enough to save them. And Erica Bree Rosenblum of the University of California, Berkeley, points to logistical challenges of trying to use the discovery, such as through widespread treatment of endangered populations. “I tend to be skeptical about whether it will be easy to translate results from lab studies into direct conservation application.”

http://www.sciencemag.org/  Science Magazine

http://news.sciencemag.org/biology/2014/07/cure-plague-frogs?rss=1 Science Magazine

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* Livestock gut microbes contributing to greenhouse gas emission

“Increased to levels unprecedented” is how the Intergovernmental Panel on Climate Change (IPCC) described the rise of carbon dioxide, methane and nitrous oxide emissions in their report on the physical science basis of climate change in 2013. According to the US Environmental Protection Agency (EPA), the atmospheric concentration of methane, a greenhouse gas some 28 times more potent than carbon dioxide has been steadily growing since the 18th century and has now increased by 50 percent compared to pre-industrial levels, exceeding 1,800 parts per billion. The EPA attributes one-fifth of methane emissions to livestock such as cattle, sheep and other ruminants. In fact, ruminant livestock are the single largest source of methane emissions, and in a country like New Zealand (NZ), where the sheep outnumber people 7 to 1, that’s a big deal. However, not all ruminants are equal when it comes to greenhouse gas emissions. It turns out that the amount of methane produced varies substantially across individual animals of the same ruminant species. To find out why this is so, a team of researchers led by the US Department of Energy Joint Genome Institute (DOE JGI) deployed high throughput DNA sequencing and specialized analysis techniques to explore the contents of the rumens of sheep in collaboration with NZ’s AgResearch Limited to see what role ruminant “microbiomes” (the microbes living in the rumen) play in this process. The study was published online June 6, 2014 in Genome Research. “We wanted to understand why some sheep produce a lot and some produce little methane,” said DOE JGI Director Eddy Rubin. “The study shows that it is purely the microbiota responsible for the difference.”

To learn why the amount of methane that ruminants produce varies, the researchers took advantage of a large sheep screening and breeding program in NZ that aims to breed low methane-emitting ruminants without impacting other traits such as reproduction and wool and meat quality. The team measured the methane yields from a cohort of 22 sheep, and from this group, they selected four sheep with the lowest methane emissions, four sheep with the highest emissions and two sheep with intermediate emission levels. Rumen metagenome DNA samples collected on two occasions from the 10 sheep were sequenced at the DOE JGI, generating 50 billion bases of data each. “The deep sequencing study contributes to this breeding program by defining the microbial contribution to the methane trait, which can be used in addition to methane measurements to assist in animal selection,” said senior scientist Graeme Attwood of AgResearch Limited, a senior author on the paper. The team then checked to see if there was a correlation between the proportions of methanogens in the eight sheep with the highest and lowest recorded methane emissions. In sheep with low methane emissions, they found elevated levels of one particular species of methanogen (Methanosphaera) while sheep with high methane emissions had elevated levels of another group of methanogens (Methanobrevibacter gottschalkii). Exploring further, the team then identified a methane-producing pathway and three variants of a gene encoding an important methane-forming reaction that were involved in elevated methane yields. While the overall changes to the methane-producing microbial community structure and methanogen abundance across sheep were rather subtle, the team reported that the expression levels of genes involved in methane production varied more substantially across sheep, suggesting differential gene regulation, perhaps controlled by hydrogen concentration in the rumen or by variations in the dwell time of their feed.

“It’s not so much the actual composition of the microbiome that determines emission — which conventional wisdom would suggest — but mostly transcriptional regulation within the existing microbes that makes the difference, which is a concept that is relatively new in metagenomic studies,” Rubin said. The team’s findings suggest new possible targets for mitigating methane emissions at the microbiome level. Screening and breeding for low-methane producing sheep is still underway, and importantly, low-methane lines then need to be tested for stability of the trait, as well as the absence of any impacts on fertility or meat or wool production. Moreover, as Attwood notes, “there needs to be an incentive for farmers to incorporate low methane animals into their flocks, that is, achieving better performance with the low methane animals or being able to claim carbon credits. If everything went well you could expect introduction of the low methane trait to begin in three years, and for there to be slow but incremental changes to the sheep industry in subsequent years.”

http://www.sciencedaily.com/  Science daily

July 22, 2014

http://www.sciencedaily.com/releases/2014/06/140617121946.htm  Original web page at Science Daily

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Amber discovery indicates Lyme disease is older than human race

The findings were made by researchers from Oregon State University, who studied 15-20 million-year-old amber from the Dominican Republic that offer the oldest fossil evidence ever found of Borrelia, a type of spirochete-like bacteria that to this day causes Lyme disease. They were published in the journal Historical Biology. In a related study, published in Cretaceous Research, OSU scientists announced the first fossil record of Rickettsial-like cells, a bacteria that can cause various types of spotted fever. Those fossils from Myanmar were found in ticks about 100 million years old. As summer arrives and millions of people head for the outdoors, it’s worth considering that these tick-borne diseases may be far more common than has been historically appreciated, and they’ve been around for a long, long time. “Ticks and the bacteria they carry are very opportunistic,” said George Poinar, Jr., a professor emeritus in the Department of Integrative Biology of the OSU College of Science, and one of the world’s leading experts on plant and animal life forms found preserved in amber. “They are very efficient at maintaining populations of microbes in their tissues, and can infect mammals, birds, reptiles and other animals. “In the United States, Europe and Asia, ticks are a more important insect vector of disease than mosquitos,” Poinar said. “They can carry bacteria that cause a wide range of diseases, affect many different animal species, and often are not even understood or recognized by doctors. “It’s likely that many ailments in human history for which doctors had no explanation have been caused by tick-borne disease.” Lyme disease is a perfect example. It can cause problems with joints, the heart and central nervous system, but researchers didn’t even know it existed until 1975. If recognized early and treated with antibiotics, it can be cured. But it’s often mistaken for other health conditions. And surging deer populations in many areas are causing a rapid increase in Lyme disease — the confirmed and probable cases of Lyme disease in Nova Scotia nearly tripled in 2013 over the previous year.

The new research shows these problems with tick-borne disease have been around for millions of years. Bacteria are an ancient group that date back about 3.6 billion years, almost as old as the planet itself. As soft-bodied organisms they are rarely preserved in the fossil record, but an exception is amber, which begins as a free-flowing tree sap that traps and preserves material in exquisite detail as it slowly turns into a semi-precious mineral. A series of four ticks from Dominican amber were analyzed in this study, revealing a large population of spirochete-like cells that most closely resemble those of the present-day Borrelia species. In a separate report, Poinar found cells that resemble Rickettsia bacteria, the cause of Rocky Mountain spotted fever and related illnesses. This is the oldest fossil evidence of ticks associated with such bacteria. In 30 years of studying diseases revealed in the fossil record, Poinar has documented the ancient presence of such diseases as malaria, leishmania, and others. Evidence suggests that dinosaurs could have been infected with Rickettsial pathogens. Humans have probably been getting diseases, including Lyme disease, from tick-borne bacteria as long as there have been humans, Poinar said. The oldest documented case is the Tyrolean iceman, a 5,300-year-old mummy found in a glacier in the Italian Alps. “Before he was frozen in the glacier, the iceman was probably already in misery from Lyme disease,” Poinar said. “He had a lot of health problems and was really a mess.”

http://www.sciencedaily.com/ Science Daily

July 8, 2014

http://www.sciencedaily.com/releases/2014/05/140529142538.htm  Original web page at Science Daily

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* Antibiotic resistance genes are essentially everywhere

The largest metagenomic search for antibiotic resistance genes in the DNA sequences of microbial communities from around the globe has found that bacteria carrying those vexing genes turn up everywhere in nature that scientists look for them. The findings reported in the Cell Press journal Current Biology on May 8 add to evidence showing just how common and abundant those resistance genes really are in natural environments. This big-picture, ecological view on a growing healthcare concern emphasizes the important relationship between antibiotic resistance in the clinic and environmental microbiology, the researchers say. “While the environment is known to harbor antibiotic-resistant strains of bacteria, as proven by many preceding studies, we did not really know the extent of their abundance,” says Joseph Nesme of the Université de Lyon in France. “The fact that we were able to detect antibiotic resistance genes at relatively important abundance in every environment tested is certainly our most striking result.” The researchers, including Nesme and senior author of the study Pascal Simonet, took advantage of the ever-growing reams of existing next-generation sequencing data that are freely available in public repositories together with information about antibiotic resistance genes found in pathogens infecting patients in the clinic. “Our strategy was simply to use all these pre-existing data and combine them to answer more precisely the question of antibiotic resistance prevalence in the environment,” Nesme says.

The scientists’ analyses detected antibiotic resistance gene determinants in all 71 environments represented in the public data, including soil, oceans, and human feces. Samples collected from soil contained the most diverse pool of resistance genes, the authors found. The most common types of resistance uncovered were efflux pumps and other genes conferring resistance to vancomycin, tetracycline, or beta-lactam antibiotics, which are in common use in veterinary and human healthcare. All this, and Simonet says they know that today’s technologies are still unable to capture all of the diversity present in the environment. In other words, we’re still missing part of the picture. There is a very good reason microbes would be armed with antibiotic resistance genes, the researchers explain. After all, most antibiotics used in medicine are isolated from soil microorganisms, such as bacteria or fungi, in the first place. That means that the resistance genes were available long before humans put antibiotic drugs into use. Bacteria lacking them to start with can simply borrow them (via horizontal transfer of genes) from those that are better equipped. Nesme and Simonet say the new findings should come as a plea for a broader ecological perspective on the antibiotic resistance problem. “It is only with more knowledge on antibiotic resistance dissemination — from the environment to pathogens in the clinic and leading to antibiotic treatment failure rates — that we will be able to produce more sustainable antibiotic drugs,” Nesme says.

http://www.sciencedaily.com/  Science Daily

June 10, 2014

http://www.sciencedaily.com/releases/2014/05/140508121347.htm Original web page at Science Daily

 

 

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* Humans and companion animals harbor the same types of MRSA infections

“Our study demonstrates that humans and companion animals readily exchange and share MRSA bacteria from the same population,” says senior author Mark Holmes, senior lecturer in preventive veterinary medicine at the University of Cambridge in England. MRSA naturally lives on the skin and also causes difficult-to-treat infections in humans and animals. “It also furthers the ‘one health’ view of infectious diseases that the pathogens infecting both humans and animals are intrinsically linked, and provides evidence that antibiotic usage in animal medicine is shaping the population of a major human pathogen.” Holmes and colleagues sequenced the genomes of 46 MRSA samples from cats and dogs, collected between August 2003 and August 2007 from two large veterinary hospitals and several smaller veterinary practices throughout the United Kingdom. The samples were found to be similar to those associated with MRSA strains in humans, with most coming from wound infections or skin and soft tissue infections. Additional samples were from the animals’ urine; cerebrospinal fluid; nasal wash or discharge; and bloodstream, heart valve or joint infections. Comparing the samples to a global collection of human MRSA samples sequenced as part of other studies and evaluating the evolution of the bacteria, the investigators found that all animal infections fell in the same family: Epidemic MRSA 15 (EMRSA-15) (sequence type ST22), a common strain of MRSA first detected in the United Kingdom in the 1990s that spread throughout Europe. The bacteria were interspersed throughout the EMRSA-15 genetic family tree. Nearly all samples were genetically similar to human bacteria, and their place in the family tree showed that the companion animal bacteria most likely originated in humans.

Researchers also observed that samples from the same veterinary hospitals clustered together genetically, suggesting that as in human hospitals, MRSA can be readily transmitted in veterinary hospital settings. “It’s a reminder that constant vigilance and high levels of hygiene are just as important when treating cats and dogs as with humans,” Holmes says. Analysis of the genomes showed very little genetic discrimination between bacteria samples from humans and animals, indicating that the MRSA from cats and dogs had not undergone extensive adaptation to the companion animals, suggesting this type of MRSA has a broad host range. But the animal MRSA were significantly less likely than those from humans to have resistance to the antibiotic erythromycin, used rarely in English veterinary practices. Instead, these MRSA from animals were more likely to contain mutations making them resistant to the antibiotic clindamycin, used widely in veterinary medicine in the United Kingdom. “MRSA infection in cats and dogs is still extremely rare,” Holmes says. “There is very little risk of owners getting ill from their pets.” In addition, he says, healthy pets are not likely to pick up MRSA from their human companions but if a pet already is ill or its health is severely compromised, MRSA patients should inform their pets’ veterinarians.

http://www.sciencedaily.com/  Science Daily

June 10, 2014

http://www.sciencedaily.com/releases/2014/05/140513092533.htm  Original web page at Science Daily

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Vet study reveals Salmonella’s hideout strategy

A study led by researchers in the University of Pennsylvania’s School of Veterinary Medicine now reveals how some Salmonella bacteria hide from the immune system, allowing them to persist and cause systemic infection. The findings could help researchers craft a more effective vaccine against Salmonella. Igor Brodsky, an assistant professor in Penn Vet’s Department of Pathobiology, was senior author on the paper, which was published in the Journal of Experimental Medicine. Co-authors included Penn Vet postdoctoral associates Meghan A. Wynosky-Dolfi and Patrick J. Doonan, Ph.D. students Naomi H. Philip, Erin E. Zwack and Amber M. Riblett and department colleague Bruce D. Freedman. The Penn researchers collaborated with Till Strowig and Richard A. Flavell of Yale University School of Medicine and Maya C. Poffenberger, Daina Avizonis and Russell G. Jones of McGill University. “Many of the same signals that are present in harmless bacteria are also present in pathogenic bacteria,” Brodsky said. “One of the big unanswered questions is how does the innate immune system distinguish between the two? And, conversely, how have pathogenic bacteria evolved to get around the immune response?” The Penn study addresses both questions, focusing on a component of the innate immune response called the inflammasome. Consisting of a complex of proteins that triggers the release of signaling molecules, the inflammasome serves to recruit other components of the immune system that can fight off the pathogen. “We hypothesized that during the systemic phase of disease, Salmonella would have some way of avoiding inflammasome activation,” Brodsky said. To identify the mechanism by which the bacteria might do this, Brodsky’s team made a library of Salmonella mutants, looking for those that might be involved in the evasion strategy. Among the 18 genes they pinpointed were four that had been previously noted to have a role in enabling Salmonella strains to cause long-term, chronic infections.

“That was interesting because it suggested that at least a subset of those genes that might be important for long-term infection might be involved in evading or suppressing the inflammasome response,” Brodsky said. They trained their attention on one of these four, the gene that encodes the enzyme aconitase. Aconitase, which converts citrate to isocitrate, is a key component in the metabolic process known as the citric acid or Krebs cycle. This cycle is used by all oxygen-breathing organisms to convert sugar into energy and to produce important molecules for cell growth. When the aconitase gene was mutated, the inflammasome known as NLRP3 was highly activated, leading researchers to believe that the normal version of aconitase might do the opposite, inhibiting the inflammasome. Moreover, when the researchers infected mice with a strain of Salmonella that had a mutated version of aconitase, the rodents were able to clear the infection, likely due to the inflammasome being activated. This infection led to increased levels of inflammation in the mice’s tissues. The Penn-led team also wanted to see whether other components of the citric acid cycle might be involved in inflammasome activation. They found that mutating Salmonella genes that encode two other players in the cycle, the enzymes isocitrate dehydrogenase and isocitrate lyase, also led to higher activation of the NLRP3 inflammasome. In their normal state, these enzymes break down citrate. Thus the study’s results point to the possibility that the immune system may activate the inflammasome in response to the presence of citrate or some byproduct of citrate. Supporting this idea, the researchers found that Salmonella strains lacking the enzyme citrate synthase, which produces citrate, led to a reduced inflammatory response. “We think bacteria might be exporting citrate because it would otherwise prevent the bacteria from growing,” Brodsky said. “It’s possible that the export of citrate might be triggering the inflammatory response. Our work fits into this emerging idea that bacterial metabolites might be recognized by various components of the immune system for the purpose of either negatively or positively regulating immune responses.” The scientists believe that it’s possible that host cells put together two pieces of information to trigger an immune response, first recognizing signaling of a Toll-like receptor, which responds to structures that are common across many microbes, and then sensing bacterial products, like elevated levels of citrate, being produced inside the cell itself. Brodsky and colleagues are now working to develop a chicken vaccine based on an attenuated strain of Salmonella that would trigger both “arms” of the inflammatory response, possibly involving an aconitase mutant. Such a vaccine would ideally more closely replicate a natural infection, protecting the animals against infection. “We get Salmonella from chickens that are chronically infected,” Brodsky said, “so, if you could prevent or limit chronic infection of chickens, that would be a nice way to limit Salmonella in the food supply.”

http://www.sciencedaily.com/  Science Daily

June 10, 2014

http://www.sciencedaily.com/releases/2014/05/140515163834.htm  Original web page at Science Daily

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* WHO warns against ‘post-antibiotic’ era

Methicillin-resistant Staphylococcus aureus (MRSA) is now invulnerable to many antibiotics. The ‘post-antibiotic’ era is near, according to a report released today by the World Health Organization (WHO). The decreasing effectiveness of antibiotics and other antimicrobial agents is a global problem, and a surveillance system should be established to monitor it, the group says. There is nothing hopeful in the WHO’s report, which pulls together data from 129 member states to show extensive resistance to antimicrobial agents in every region of the world. Overuse of antibiotics in agriculture — to promote livestock growth — and in hospitals quickly leads to proliferation of drug-resistant bacteria, which then spread via human travel and poor sanitation practices. “A post-antibiotic era — in which common infections and minor injuries can kill — far from being an apocalyptic fantasy, is instead a very real possibility for the twenty-first century,” writes Keiji Fukuda, WHO assistant director-general for health security, in a foreword to the report. Perhaps the most worrying trend is the spread of resistance to carbapenems, the ‘antibiotics of last resort’, says Timothy Walsh, a medical microbiologist at Cardiff University, UK, who was an adviser for the report. “That’s taken us by surprise,” he says. “All of us are rather like rabbits in front of the headlights in how quickly this has taken off.” The report finds that, in some areas of the world, more than half the infections caused by one major category of bacteria — Gram-negative, which includes Escherichia coli and Klebsiella pneumoniae — involve species resistant to carbapenem drugs. There are few if any replacements for carbapenems in development, says Elizabeth Jungman, director of drug safety and innovation at the Pew Charitable Trusts in Washington DC. Companies lack economic incentives to develop new antibiotics, she says, and researchers have found it difficult to find new ways to get Gram-negative bacteria to take up antibiotics.

Ultimately, the report’s most surprising finding may be the lack of global data on antimicrobial resistance. “Despite the fact we’ve known the potential of this going cataclysmic for ten years, as a global unit we haven’t managed to get our act together,” says Walsh. Just 22 of the 129 WHO member states that contributed to the report had data on the nine antibiotic–bacteria pairs of greatest concern. Although the report calls for the establishment of a global monitoring network, it is unlikely that any extra money is forthcoming. “It’s a huge problem and I’m not sure the resources are available,” says Keith Klugman, an epidemiologist at the Bill and Melinda Gates Foundation in Seattle, Washington.

Nature doi:10.1038/nature.2014.15135

http://www.nature.com/news/index.html  Nature

May 27, 2014

http://www.nature.com/news/who-warns-against-post-antibiotic-era-1.15135 Original web page at Nature

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Cougars’ diverse diet helped them survive the mass extinction that wiped out the saber-tooth cat, American lion

Cougars may have survived the mass extinction that took place about 12,000 years ago because they were not particular about what they ate, unlike their more finicky cousins — the saber-tooth cat and American lion. Both perished along with the woolly mammoth and many of the other supersized mammals that walked Earth during the late Pleistocene. That is the conclusion of a new analysis of the microscopic wear marks on the teeth of cougars, saber-tooth cats and American lions described in the April 23 issue of the journal Biology Letters. “Before the Late Pleistocene extinction six species of large cats roamed the plains and forests of North America. Only two — the cougar and jaguar — survived. The goal of our study was to examine the possibility that dietary factors can explain the cougar’s survival,” said Larisa R.G. DeSantis, assistant professor of earth and environmental sciences at VanderbiltUniversity, who co-authored the study with Ryan Haupt at the University of Wyoming. For their investigation, DeSantis and Haupt employed a new technique called dental microwear texture analysis. DMTA uses a confocal microscope to produce a three-dimensional image of the surface of a tooth. The image is then analyzed for microscopic wear patterns. The analysis of the teeth of modern carnivores, including hyenas, cheetahs and lions has established that the meals an animal consumes during the last few weeks of its life leave telltale marks. Chowing down on red meat, for example, produces small parallel scratches while chomping on bones adds larger, deeper pits. The researchers analyzed the teeth of 50 fossil and modern cougars, and compared them with the teeth of saber-tooth cats and American lions excavated from the La Brea Tar Pits in Los Angeles and the teeth of modern African carnivores including cheetahs, lions and hyenas.

Previously, DeSantis and others found that the dental wear patterns of the extinct American lions closely resembled those of modern cheetahs, which are extremely finicky eaters that mostly consume tender meat and rarely gnaw on bones. Saber-tooth cats were instead similar to African lions and chewed on both flesh and bone. Among the La Brea cougars the researchers found significantly greater variation between individuals than they did in the other large cats, including saber-toothed cats. Some of the cougars show wear patterns similar to those of the finicky eaters but on others they found wear patterns closer to those of modern hyenas, which consume almost the entire body of their prey, bones included. “This suggests that the Pleistocene cougars had a ‘more generalized’ dietary behavior,” DeSantis said. “Specifically, they likely killed and often fully consumed their prey, more so than the large cats that went extinct.” This is consistent with the dietary behavior and dental wear patterns of modern cougars, which are opportunistic predators and scavengers of abandoned carrion and fully consume the carcasses of small and medium-sized prey, a “variable dietary behavior that may have actually been a key to their survival.”

http://www.sciencedaily.com/  Science Daily

May 13, 2014

http://www.sciencedaily.com/releases/2014/04/140422202047.htm  Original web page at Science Daily

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* Cow manure harbors diverse new antibiotic resistance genes

Manure from dairy cows, which is commonly used as a farm soil fertilizer, contains a surprising number of newly identified antibiotic resistance genes from the cows’ gut bacteria. The findings, reported in mBio® the online open-access journal of the American Society for Microbiology, hints that cow manure is a potential source of new types of antibiotic resistance genes that transfer to bacteria in the soils where food is grown. Thousands of antibiotic resistance (AR) genes have already been identified, but the vast majority of them don’t pose a problem when found in harmless bacteria. The real worry is when these genes appear in the types of pathogenic bacteria that cause food-borne illnesses or hospital infections. “Since there is a connection between AR genes found in environmental bacteria and bacteria in hospitals, we wanted to know what kind of bacteria are released into the environment via this route,” of manure fertilization, says Fabienne Wichmann, lead study author and former postdoctoral researcher at Yale University in New Haven, Connecticut. Farmers use raw or composted cow manure on some vegetable crops, which could lead to a scenario where residual manure bacteria might cling to produce and they or their genes might move to the human ecosystem. “Is this a route for movement of these genes from the barn to the table?” asks Jo Handelsman, senior study author and microbiologist at Yale. The first step toward an answer was surveying which AR genes are present in cow manure. Handelsman’s team used a powerful screening-plus-sequencing approach to identify 80 unique and functional AR genes. The genes made a laboratory strain of Escherichia coli bacteria resistant to one of four types of antibiotics — beta-lactams (like penicillin), aminoglycosides (like kanamycin), tetracycline, or chloramphenicol.

Roughly 75% of the 80 AR genes had sequences that were only distantly related to AR genes already discovered. The team also found an entire new family of AR genes that confer resistance to chloramphenicol antibiotics, which are commonly used to treat respiratory infections in livestock. “The diversity of genes we found is remarkable in itself considering the small set of five manure samples,” says Handelsman, who is also a Howard Hughes Medical Institute Professor. “But also, these are evolutionarily distant from the genes we already have in the genetic databases, which largely represent AR genes we see in the clinic.” That might signal good news that AR genes from cow gut bacteria are not currently causing problems for human patients. But, Wichmann points out, another possibility is that “cow manure harbors an unprecedented reservoir of AR genes” that could be next to move into humans. “This is just the first in a sequence of studies — starting in the barn, moving to the soil and food on the table and then ending up in the clinic — to find out whether these genes have the potential to move in that direction,” says Handelsman. AR genes can enter the human ecosystem by two routes — either the bacteria that contain them colonize humans, or the genes are transferred through a process called horizontal gene transfer to other bacteria that colonize humans. Research has already shown that bacteria are transferred from farm animals to their human caretakers. Gene transfer enables genes to jump between microorganisms that are not related, and it occurs in most environments that host bacteria. Some manure bacteria might be pathogenic to humans, so if they acquire antibiotic resistance, they could pose a problem. Alternatively, benign bacteria in manure might transfer resistance genes to pathogens at any point along the path — in manure, soil, food, or humans. “We’re hoping this study will open up a larger field of surveillance, to start looking at new types of resistance before they show up in the clinic,” says Handelsman.

http://www.sciencedaily.com/ Science Daily

May 13, 2014

http://www.sciencedaily.com/releases/2014/04/140422084928.htm Original web page at Science Daily

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* New light shed on key bacterial immune system

New insights into a surprisingly flexible immune system present in bacteria for combating viruses and other foreign DNA invaders have been revealed by researchers from New Zealand’s University of Otago and the Netherlands. A team led by Dr Peter Fineran of the Department of Microbiology and Immunology are studying the genetic basis of adaptive immunity in bacteria that cause potato ‘soft rot’ and in E. coli bacteria. Through their recent collaboration they have found that these bacterial immune systems are much more robust and responsive than previously thought. Their latest findings, which appear in the leading US journal PNAS, have implications for improving our understanding of bacterial evolution, including the spread of antibiotic resistance genes. The researchers are investigating an adaptive immune system, termed CRISPR-Cas, which is found in half of all bacterial species and in almost all single-celled microbes in the archaea domain. CRISPR-Cas’s role in providing immunity was only discovered in the past decade. The system creates a genetic memory of specific past infections by viruses and plasmids, which are small mobile DNA molecules that can move between organisms. Dr Fineran says the system steals samples of the invader’s genetic material and stores them in a memory bank so it can immediately recognize future exposures and neutralize the attack. It can store up to 600 samples and can also pass on these memories to subsequent generations of bacteria. It had been thought that the system had an Achilles heel because invaders that had acquired too many mutations could no longer be recognized as they did not match the stored sample closely enough.

“What we have now discovered is that while the viruses and plasmids can evade direct recognition by acquiring multiple mutations, the system is primed to quickly generate a new immunity by grabbing a new sample of the mutated genetic material.” “It’s a remarkably flexible and robust immune system for such simple single-celled organisms.” Dr Fineran says the system reflected the ancient and continuing co-evolutionary arms race between bacteria on one side, and viruses and plasmids on the other. Viral infections of bacteria also exert a powerful yet invisible effect on the entire planet, says Dr Fineran. “Their silent but vast and ongoing war underpins everything from how global nutrient cycles — which rely on bacteria to produce half of Earth’s biomass — operate, to how human pathogens evolve,” he says. “For example, the bacteria that cause cholera and diphtheria have been infected by viruses that provide genes coding for toxins, which converted these bacteria into significant human pathogens.” Plasmids are also key players in moving antibiotic resistance genes between different bacterial species. “So, discovering more about exactly how bacterial immune systems combat plasmid transfer and acquisition is of considerable interest,” he says.

http://www.sciencedaily.com/ Science Daily

April 29, 2014

http://www.sciencedaily.com/releases/2014/04/140407153917.htm  Original web page at Science Daily

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Spread of antibiotic resistance understood by unravelling bacterial secretion system

The system that allows the sharing of genetic material between bacteria — and therefore the spread of antibiotic resistance — has been uncovered by a team of scientists at Birkbeck, University of London and UCL. The study, published in Nature, reveals the mechanism of bacterial type IV secretion, which bacteria use to move substances across their cell wall. As type IV secretion can distribute genetic material between bacteria, notably antibiotic resistance genes, the mechanism is directly responsible for the spread of antibiotic resistance in hospital settings. It also plays a crucial role in secreting toxins in infections — causing ulcers, whooping cough, or severe forms of pneumonia such as Legionnaires’ disease. The work, led by Professor Waksman at the Institute of Structural and Molecular Biology (a joint Birkbeck/UCL Institute) and funded by the Wellcome Trust, revealed that the type IV secretion system differs substantially from other bacterial secretion systems, in both its molecular structure and the mechanism for secretion. Professor Waksman said: “This work is a veritable tour de force. The entire complex is absolutely huge and its structure is unprecedented. It is the type of work which is ground-breaking and will provide an entirely new direction to the field. Next, we need to understand how bacteria use this structure to get a movie of how antibiotics resistance genes are moved around.”

Using electron microscopy the team were able to reconstruct the system as observed in the bacteria E. coli. They saw that the mechanism consists of two separate complexes, one in the outer membrane of the cell, and the other in the inner membrane, which are connected by a stalk-like structure that crosses the periplasm — the space between the two membranes. The complexes at both the inner and outer membranes form pores in the membrane, via which substances can be secreted. Understanding the structure of the secretion system will help scientists uncover the mechanism by which it moves substances across the inner and outer membranes. It could eventually help scientists develop new tools for the genetic modification of human cells, as the bacteria could act as a carrier for genetic material, which could then be secreted into cells. Professor Waksman said: “Understanding bacteria’s secretion system could help design new compounds able to stop the secretion process, thereby stopping the spread of antibiotics resistance genes. Given that antibiotics resistance has become so widespread and represents a grave threat to human health, the work could have a considerable impact for future research in the field of antimicrobials.”

http://www.sciencedaily.com/  Science Daily

April 1, 2014

http://www.sciencedaily.com/releases/2014/03/140309150544.htm  Original web page at Science Daily

 

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Essential factor for Lyme disease transmission identified

Borrelia burgdorferi, the bacterium that causes Lyme disease, hitchhikes in ticks for dissemination to mammalian hosts–including humans. An article in the 19 December issue of PLOS Pathogens identifies HrpA, an RNA helicase, as a crucial player in the transmission from ticks to mammals. George Chaconas, from the University of Calgary, Canada, and a member of the university’s Snyder Institute for Chronic Diseases, and colleagues had previously identified HrpA as a modulator of B. burgdorferi protein expression. For this study, Chaconas’ group joined forces with Justin Radolf and Melissa Caimano from the University of Connecticut Health Center, USA, to analyze the molecular function of the HrpA protein and further explore its role in the bacterium’s complicated life cycle, in particular for transmission of the pathogen. Its DNA sequence suggests that HrpA is an RNA helicase, a protein that can harvest energy from the cell’s stores, use it to unwind RNA, and so regulate translation of RNA into protein. Most bacteria have several putative helicases, including one from the HrpA family, but nothing was known about the actual HrpA function from other species. HrpA is the only putative RNA helicase in B. burgdorferi, and the scientists found that it indeed possesses the multiple activities characteristic of a helicase: it can bind to RNA and use its ATPase activity to harvest energy, which in turn is used to unwind the RNA strand. They also showed that these activities are involved in the regulation of target RNAs.

When the scientists tested whether mutant B. burgdorferi that lacked the hrpA gene could infect mice, they found that the mutant bacteria could not. For this experiment, the scientists injected normal or mutant bacteria directly into mice, and subsequently tested mouse blood, skin, bladder, or joint tissue for the presence of bacteria. Normal bacteria could be recovered from all tissues after a week and up to 4 weeks post injection, but mutant bacteria were undetectable even after one week, suggesting that they were unable to survive or multiply in the mammalian host. HrpA-deficient bacteria were also unable to infect mice using the natural route, i.e. via a bite from an infected tick. This was not because the mutant bacteria were unable to grow or survive in the ticks. Rather the mutants could not exit the tick midgut or enter the salivary glands, where Borrelia needs to be for successful transmission during feeding; even right after the engorged ticks fell off, mutant bacteria were not detectable in the mouse skin around the attachment site. The authors say, “We now know that HrpA is involved in both parts of the B. burgdorferi lifecycle: animal infection and tick transmission, making it a very important protein in B. burgdorferi gene regulation and establishing gene regulation through an RNA helicase as an important regulatory pathway in the Lyme spirochete.”

Science Daily
January 21, 2014

Original web page at Science Daily

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New staph vaccine shows promise in rabbits

Staph infections pose one of the most serious microbial threats to people. Yet efforts to prevent them with vaccines have consistently failed, even as the bacterium responsible becomes increasingly resistant to antibiotics. Now, an experimental vaccine with novel ingredients has proven highly protective in rabbits against staph pneumonia, one of the most dangerous outcomes of the bacterial infection. The inventor hopes to take it into human studies soon. Staphylococcus aureus is thought to colonize the noses of about one-third of humans at any given time, but it also causes about half a million hospitalizations and 20,000 deaths each year in the United States. Strains of the bacterium that are resistant to most antibiotics, particularly varieties dubbed MRSA (methicillin-resistant S. aureus), have become a major danger, highlighting the need for a vaccine against the microbe. However, a series of vaccines has suffered dramatic failures in the past few years. A candidate from NABI Biopharmaceuticals of Rockville, Maryland, flopped in two trials among dialysis patients in the United States, and a Merck vaccine was abandoned in 2012 after millions of dollars were spent testing it. Microbiologist Patrick Schlievert of the University of Iowa in Iowa City believes that pharmaceutical companies have been going about a vaccine the wrong way. Previous staph vaccines attempted to offer protection by including proteins or carbohydrates contained in the natural capsule that surrounds the staph bacterium’s cell wall and enables it to elude the immune system. This approach succeeded for vaccines against other bacteria like Haemophilus influenzae type B and Streptococcus pneumoniae, but did not work with S. aureus. Other vaccines that failed included single proteins from the cell wall of the S. aureus bacterium.

In an article published online this month in The Journal of Infectious Diseases, Schlievert and colleagues took a new approach. The strategy involves an entirely different class of substances taken from the S. aureus bacterium. These included so-called superantigens and cytolysins—proteins that are produced by S. aureus‘s internal structures and play major roles in the illnesses caused by the bacterium. The researchers vaccinated 88 rabbits, split into several groups, with different combinations of the substances. All but two of the animals survived when S. aureus organisms were sprayed into their lungs. Of 88 unvaccinated rabbits exposed to the same bacteria, only one survived. In another experiment, the researchers found evidence that typical vaccines may actually make those immunized more vulnerable to a staph infection. They vaccinated five rabbits with neutralized proteins from the cell surface of S. aureus—the type of substances used in most previous and current vaccine candidates. All five of those animals died within 6 hours of exposure to a common MRSA strain; in contrast, five nonvaccinated animals survived at least 4 days postexposure. The studies’ counterintuitive findings are in agreement with prior research by Schlievert indicating that cell-surface antigens, which are frequently used to create bacterial vaccines, are a poor choice for preventives aimed at S. aureus because of the organism’s particular way of causing disease, he says.

“One of the ways staph works is by creating aggregations of bacteria that block blood vessels and airwaves,” Schlievert says. Consequently, vaccines that stimulate the creation of antibodies to cell-surface antigens may be more dangerous than no vaccine at all, he says, because in binding with those antigens, they create molecular complexes that intensify the clumping phenomenon. Schlievert also believes that his results are more relevant to humans than other animal studies with staph vaccines. While other researchers have tested S. aureus vaccines in mice and nonhuman primates, he contends that the immune system of the rabbit is more similar to that of humans, at least when it comes to how it responds to this bacterium. Staph researchers say that Schlievert’s new study is provocative but not entirely convincing. “He shows that superantigens are more protective than surface antigens—in rabbits undergoing experimental infection,” says Robert Daum, pediatrician and microbiologist at the University of Chicago in Illinois. It remains to be seen whether the same holds true in humans, he says. “Lots of people have been using surface antigens in vaccines for a long time. It hasn’t been a great success story, but there are few data that suggest the vaccines make it worse.”

Daum considers the suggestion that rabbits were better than mice as experimental animals to be intriguing. It is true that mice are not especially good models in S. aureus, he says, but on the other hand the rodents are inexpensive and well-studied. “If he’s right that rabbits are a better model, it will roil the field, because they are much more expensive and difficult to work with. To my mind, rabbits haven’t been studied enough yet to conclude that they are better.” Most of the data to this effect comes from Schlievert’s group. Daum argues that failures in previous vaccines have shown that defeating S. aureus requires more basic science before new vaccine antigens are trotted out. Too little is understood about the human immune response to S. aureus, which can cause deadly infections, yet often lives in and on us without causing disease at all. “We don’t want to eliminate all staph. We want to eliminate the nasty ones that cause disease. So we need to understand the immunology, and we’re not there yet,” Daum says. Schlievert says that unpublished data from his group show that his superantigen-based vaccines also prevent skin infections in rabbits. He next plans to ask the Food and Drug Administration to allow him to conduct basic human safety studies of the vaccine.

ScienceNow
January 21, 2014

Original web page at ScienceNow

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Quantitative approaches provide new perspective on development of antibiotic resistance

Using quantitative models of bacterial growth, a team of UC San Diego biophysicists has discovered the bizarre way by which antibiotic resistance allows bacteria to multiply in the presence of antibiotics, a growing health problem in hospitals and nursing homes across the United States. Two months ago, the Centers for Disease Control and Prevention issued a sobering report estimating that antibiotic-resistant bacteria last year caused more than two million illnesses and approximately 23,000 deaths in the United States. Treating these infections, the report said, added $20 billion last year to our already overburdened health care system. Many approaches are now being employed by public health officials to limit the spread of antibiotic resistance in bacteria — such as limiting the use of antibiotics in livestock, controlling prescriptions of antibiotics and developing new drugs against bacteria already resistant to conventional drug treatments. But understanding how bacteria grow and evolve drug resistance could also help stop its spread by allowing scientists to target the process of evolution itself. “Understanding how bacteria harboring antibiotic resistance grow in the presence of antibiotics is critical for predicting the spread and evolution of drug resistance,” the UC San Diego scientists say in an article published in the November 29 issue of the journal Science.

In their study, the researchers found that the expression of antibiotic resistance genes in strains of the model bacterium E. coli depends on a complex relationship between the bacterial colony’s growth status and the effectiveness of the resistance mechanism. “In the course of developing complete resistance to a drug, a strain of bacteria often first acquires a mechanism with very limited efficacy,” says Terry Hwa, a professor of physics and biology who headed the research effort. “While much effort has been spent elucidating individually how a drug inhibits bacterial growth and how a resistance mechanism neutralizes the action of a drug, little is known previously about how the two play off of each other during the critical phase where drug resistance evolves towards full strength.” According to Hwa, the interaction between drug and drug-resistance is complex because the degree of drug resistance expressed in a bacterium depends on its state of growth, which in turn depends on the efficacy of drug, with the latter depending on the expression of drug resistance itself. For a class of common drugs, the researchers realized that this chain of circular relations acted effectively to promote the efficacy of drug resistance for an intermediate range of drug doses.

The use of predictive quantitative models was instrumental in guiding the researchers to formulate critical experiments to dissect this complexity. In their experiments, E. coli cells possessing varying degrees of resistance to an antibiotic were grown in carefully controlled environments kept at different drug doses in “microfluidic” devices — which permitted the researchers to manipulate tiny amounts of fluid and allowed them to continuously observe the individual cells. Hwa and his team found a range of drug doses for which genetically identical bacterial cells exhibited drastically different behaviors: while a substantial fraction of cells stopped growing despite carrying the resistance gene, other cells continued to grow at a high rate. This phenomenon, called “growth bistability,” occurred as quantitatively predicted by the researchers’ mathematical models, in terms of both the dependence on the drug dose, which is set by the environment, and on the degree of drug resistance a strain possesses, which is set by the genetic makeup of the strain and is subject to change during evolution. “Exposing this behavior generates insight into the evolution of drug resistance,” says Hwa. “With this model we can chart how resistance is picked up and evaluate quantitatively the efficacy of a drug.” However, this model has only been established for one class of drugs and one class of drug-resistance mechanisms. Hwa believes it is important to establish such predictive models for all the common drugs in pathogenic bacterial species.

“My hope,” he adds, “is to get the message out to drug companies and hospitals that there is an informative, quantitative way to look at the action of a drug on bacteria and at the consequences of using a drug on bacteria as they try to pick up resistance, and that this approach can be incorporated in both the design and evaluation of drug efficacy in clinically relevant settings. “Hwa says the principle of interaction between drug and drug-resistance is important to understand not only for the evolution of antibiotics, but also for the emergence of drug resistance in other diseases. A prominent example is the rapid emergence of cancer lines resistant to drug treatment, which underlies most failures in cancer drug therapies. While there are obviously numerous differences between the evolution of drug resistance in bacteria and in cancer, Hwa noted that the connection between the two was sufficient to motivate the Physical Science-Oncology program of the National Cancer Institute to co-sponsor this study.

Science Daily
January 7, 2014

Original web page at Science Daily

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Bacteria recycle broken DNA: Modern bacteria can add DNA from creatures long-dead to its own

From a bacteria’s perspective the environment is one big DNA waste yard. Researchers have now shown that bacteria can take up small as well as large pieces of old DNA from this scrapheap and include it in their own genome. This discovery may have major consequences – both in connection with resistance to antibiotics in hospitals and in our perception of the evolution of life itself. Our surroundings contain large amounts of strongly fragmented and damaged DNA, which is being degraded. Some of it may be thousands of years old. Laboratory experiments with microbes and various kinds of DNA have shown that bacteria take up very short and damaged DNA from the environment and passively integrate it in their own genome. Furthermore this mechanism has also been shown to work with a modern bacteria’s uptake of 43,000 years old mammoth DNA. The results are published now in the scientific journal Proceedings of the National Academy of Sciences (PNAS). The discovery of this second-hand use of old or fragmented DNA may have major future consequences. Postdoc Søren Overballe-Petersen from the Centre for GeoGenetics at the Natural History Museum of Denmark is first author on the paper and he says about the findings: “It is well-known that bacteria can take up long intact pieces of DNA but so far the assumption has been that short DNA fragments were biologically inactive. Now we have shown that this assumption was wrong. As long as you have just a tiny amount of DNA left over there is a possibility that bacteria can re-use the DNA. One consequence of this is in hospitals that have persistent problems with antibiotic resistance. In some cases they will have to start considering how to eliminate DNA remnants. So far focus has been on killing living pathogen bacteria but this is no longer enough in the cases where other bacteria afterwards can use the DNA fragments which contain the antibiotic resistance.”

The research group’s results reveal that the large reservoir of fragments and damaged DNA in the surroundings preserve the potential to change the bacteria’s genomes even after thousands of years. This is the first time a process has been described which allows cells to acquire genetic sequences from a long gone past. We call this phenomenon Anachronistic Evolution – or Second-hand Evolution. Professor Eske Willerslev from the Centre for GeoGenetics at the Natural History Museum of Denmark is the leader of the project. He says: “That DNA from dead organisms drives the evolution of living cells is in contradiction with common belief of what drives the evolution of life itself.” Furthermore old DNA is not limited to only returning microbes to earlier states. Damaged DNA can also create new combinations of already functional sequences. You can compare it to a bunch of bacteria which poke around a trash pile looking for fragments they can use. Occasionally they hit some ‘second-hand gold’, which they can use right away. At other times they run the risk of cutting themselves up. It goes both ways. This discovery has a number of consequences partially because there is a potential risk for people when pathogen bacteria or multi-resistant bacteria exchange small fragments of ‘dangerous’ DNA e.g. at hospitals, in biological waste and in waste water. In the grand perspective the bacteria’s uptake of short DNA represents a fundamental evolutionary process that only needs a growing cell consuming DNA pieces. A process that possibly is a kind of original type of gene-transfer or DNA-sharing between bacteria. The results show how genetic evolution can happen in jerks in small units. The meaning of this is great for our understanding of how microorganisms have exchanged genes through the history of life. The new results also support the theories about gene-transfer as a decisive factor in life’s early evolution. Søren Overballe-Petersen explains: “This is one of the most exciting perspectives of our discovery. Computer simulations have shown that even early bacteria on Earth had the ability to share DNA – but it was hard to see how it could happen. Now we suggest how the first bacteria exchanged DNA. It is not even a mechanism developed to this specific purpose but rather as a common process, which is a consequence of living and dying.”

Science Daily
December 10, 1013

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New vaccine against lung diseases in goats and sheep

An intranasal spray was developed using local isolated bacterium in Malaysia and it was found to provide better protection against infections by Mannheimia haemolytica bacterium than imported vaccines. Universiti Putra Malaysia has launched a new vaccine against lung or pneumonic diseases in goats and sheep that was developed and patented by its scientists. The soft launch of “STVac7,” the first intranasal spray vaccine for goats and sheep, was officiated by the Deputy Minister of Science, Technology and Innovation, Datuk Dr Abu Bakar Mohamad Diah, in a brief ceremony on 24 Oct., 2013. The vaccine was developed and tested from 1998 to 2005 by UPM scientists led by Prof Dr Mohd Zamri Saad of the Faculty of Veterinary Medicine. The other scientist involved is Dr Md. Sabri Mohd Yusoff. The patent for the STVac7 vaccine has been commercialised to Fed Tech Sdn Bhd, for RM4 million and the company has appointed Bio-Angle Vacs Sdn Bhd to manage the production and marketing of the vaccine, said the Vice Chancellor of UPM, Prof Datuk Dr Mohd Fauzi Hj Ramlan, when speaking at the ceremony. He said he is confident that the vaccine could penetrate the local and foreign market, since in Malaysia alone, there were more than 600,000 goats and sheep at present and the figure is expected to increase to 1 million by 2015, a growth rate of 12.1% as projected by the Veterinary Services Department.

The company’s collaboration with UPM would ensure that it would produce the STVac7 vaccines under Good Manufacturing Practices (GMP) since it would be using the facilities at FTU.GMP@Biotech (or FTU), a service centre under the Faculty of Biotechnology and Biomolecular Science of UPM. Dr Fauzi said FTU which was established in 1999 is now equipped with the facilities for upstream and downstream production of biotechnological products, including packaging and bottling. The FTU services centre which cost RM14 million to develop, is now serving a horde of local companies like Malaysia Agriculture HighTech, Stella Gen, Johor BioMicrobe, Pascal Biotech, MVP, One Biotech for the development and production of their biotechnology products. He said Bio-Angle Vacs Sdn Bhd and FTU were currently developing the standard operating procedure (SOP) for the mass production of the STVAC 7 vaccine using the GMP facilities of the service centre as well as product registration before marketing. “Actual production for the commercialisation of the product with the GMP certification will begin in July 2014 using the Original Equipment Concept,” Dr Fauzi said.

Meanwhile, replying to questions, Prof Dr Zamri said the STVac7 is a vaccine against mannheimiosis or respiratory diseases of goats and sheep caused by bacteria. It was developed and produced using sophisticated recombinant technology which, unlike the imported vaccines, has been demonstrated to provide protection against bacterium infection in the small ruminants like goats and sheep. “The current available vaccines against this disease are imported vaccines prepared using foreign strains. They are given via intramuscular injections. The vaccines were found to be ineffective and expensive. “Therefore, STVac7 was developed using local isolated bacterium that was found to be able to provide protection against infections by Mannheimia haemolytica bacterium A2, A7 and A9. This was proven to be better protection,” said Prof Zamri who graduated with his Doctor of Veterinary Medicine degree from UPM before obtaining his PhD from Liverpool. “The product is cheap and effective. If used according to the suggested protocol, it can reduce mortality due to this disease by more than 90%,” he added. Prof Zamri said the pneumonic diseases brought about by the bacterium usually caused a mortality rate of 30% during the rainy season and the goats and sheep farmers could benefit from the STVac7. “The product itself is ready for the market since laboratory and field tests have been completed,” he said, adding that they started their research in 1998 which ended in 2005.

Science Daily
December 10, 2013

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Smelly microbes help hyenas to communicate

Hyenas such as this one at Masai Mara National Reserve in Kenya recognize each other’s smell — or perhaps the smell of each other’s microbiome. The hordes of microbes that inhabit every nook and cranny of every animal are not just passive hitchhikers: they actively shape their hosts’ well-being and even behaviour. Now, researchers have found evidence that bacteria living in the scent glands of hyenas help to produce the smells that the animals use to identify group members and tell when females are ready to mate. Kevin Theis, a microbial ecologist at Michigan State University in East Lansing, had been studying hyena scent communication for several years when, after he gave a talk on the subject, someone asked him what part the bacteria might play. “I just said, ‘I don’t know’,” he says. He started investigating. He found that for 40 years, scientists had wondered whether smelly bacteria were involved in animals’ chemical communication. But experiments to determine which bacteria were present had been inconclusive, because the microbes had to be grown in culture, which is not possible with all bacteria. However, next-generation genetic sequencing would enable Theis to identify the microbes in a sample without having to grow them in a dish. Using this technique, Theis and his colleagues last year published a study that identified more types of bacterium living in the hyenas’ scent glands than the 15 previous studies of mammal scent glands combined. In both spotted hyenas (Crocuta crocuta) and striped hyenas (Hyaena hyaena), most of the bacteria were of a kind that ferments nutrients exuded by the skin and produces odours. “The diversity of the bacteria is enough to potentially explain the origin of these signals,” says Theis.

Now, they have found that the structure of the bacterial communities varied depending on the scent profiles of the sour, musky-smelling ‘pastes’ that the animals left on grass stalks to communicate with members of their clan. In addition, in the spotted hyenas, both the bacterial and scent profiles varied between males and females, and with the reproductive state of females — all attributes that hyenas are known to be able to infer from scent pastes. The work is published this week in Proceedings of the National Academy of Sciences. Although this work is specific to hyenas, Theis thinks that bacteria probably also have a role in chemical communication in other animals. Jerry Wells, who studies host–microbe interactions at Wageningen University in the Netherlands, agrees. He says that the work is “a striking example of how microbiota might communicate information about their hosts, and how symbiosis of microbiota can be linked to chemical signalling and animal behaviour”. And he adds that the whiff of microbes could communicate much more: “Who knows, perhaps microbiota can reveal other information about their hosts, such as health, diet, stress status and social-association patterns.”

Nature
November 26, 2013

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Gene variants in immune system pathways correlated with composition of microbes of human body

Human genes in immunity-related pathways are likely associated with the composition of an individual’s microbiome, which refers to the bacteria and other microbes that live in and on the body, scientists reported Oct. 24, at the American Society of Human Genetics 2013 annual meeting in Boston. “These genes are significantly enriched in inflammatory and immune pathways and form an interaction network highly enriched with immunity-related functions,” said Ran Blekhman, Ph.D., Assistant Professor, Department of Genetics, Cell Biology, and Development at the University of Minnesota, Minneapolis. The study is the first genome-wide and microbiome-wide investigation to identify the interactions between human genetic variation and the composition of the microbes that inhabit the human body. The skin, genital areas, mouth, and other areas of the human body, especially the intestines, are colonized by trillions of bacteria and other microorganisms. “Shifts in the composition of the species of the microbes have been associated with multiple chronic conditions, such as diabetes, inflammatory bowel disease and obesity,” noted Dr. Blekhman. Dr. Blekhman and his collaborators found evidence of genetic influences on microbiome composition at 15 body sites of 93 people surveyed. “We found in our study that genetic variation correlated with the microbiome at two levels,” he said.

At the individual level, the mathematical procedure known as principal component analysis demonstrated that genetic variation correlated with the overall structure of a person’s microbiome. At the species level, potential correlations between host genetic variation and the abundance of a single bacterial species were identified, said Dr. Blekhman, who conducted much of the research while a scientist in the lab of Andrew G. Clark, Ph.D., the Jacob Gould Schurman Professor of Population Genetics in the Department of Molecular Biology and Genetics at Cornell University, Ithaca, NY. Dr. Clark is the senior author of the abstract. To identify the bacterial species that inhabited each human body site, the researchers mined sequence data from the Human Microbiome Project (HMP), an international program to genetically catalog the microbial residents of the human body. Using a systems-level association approach, the researchers showed that variation in genes related to immune system pathways was correlated with microbiome composition in the 15 host body sites. To shed light on the evolutionary history of the symbiosis between humans and their microbiomes, the researchers analyzed sequencing data from the 1000 Genomes Project, which is designed to provide a comprehensive resource on human genetic variation.

They found that the genes in the pathways linked to the composition of an individual’s microbiome vary significantly across populations. “Moreover, many of those genes have been shown in recent studies to be under selective pressure,” said Dr. Blekhman. “The results highlight the role of host immunity in determining bacteria levels across the body and support a possible role for the microbiome in driving the evolution of bacteria-associated host genes,” he added. Dr. Blekhman is currently investigating the combined role of host genetics and the microbiome in influencing an individual’s susceptibility to such diseases as colon cancer. His goal is to unravel the interaction between host genomic variation and the gut microbiome in colon cancer incidence, evolution and therapeutic response.

Science Daily
November 12, 2013

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How a ubiquitous herpesvirus sometimes leads to cancer

You might not know it, but most of us are infected with the herpesvirus known as Epstein-Barr virus (EBV). For most of us, the virus will lead at worst to a case of infectious mononucleosis, but sometimes, and especially in some parts of the world, those viruses are found in association with cancer. Now, researchers reporting in the Cell Press journal Cell Reports on October 10 have found that the difference between a relatively harmless infection and a cancer-causing one lies at least partly in the viral strain itself. The results offer some of the first evidence for the existence of distinct EBV subtypes with very different public health risks. The researchers say that vaccination or other strategies for preventing EBV infection will need to be designed with these most pathogenic, cancer-causing strains in mind. “EBV is an important but neglected pathogen,” said Henri-Jacques Delecluse of the German Cancer Research Centre in Heidelberg, Germany. “We have made an important step in recognizing that EBV is actually a family of viruses that have different properties, some of which are very likely to cause disease. So, the consequences of being infected with EBV might be different, depending on the strain one carries.”

Delecluse and his colleagues made the discovery by sequencing the DNA of a viral strain dubbed M81 isolated from a Chinese patient with nasopharyngeal carcinoma (NPC). Their analyses revealed that M81 is highly similar to other viruses isolated from NPCs and profoundly different from Western strains in terms of its ability to infect and replicate within cells. The M81 strain can infect epithelial cells and multiply spontaneously at a very high level in all cells it infects, including B lymphocytes, the cells in which the viruses hide, the researchers report. It remains to be seen exactly how infected epithelial cells become cancerous. “Our results have made me radically change my strategy to address the problem of EBV-associated diseases,” Delecluse said. “The current view is that the virus is essentially the same all over the world and that local conditions explain the different consequences of EBV infection. We now show that the type of EBV also plays an important role. By concentrating on the potentially pathogenic EBV strains, we will soon better understand how EBV causes diseases, and this will also help in designing prevention strategies.”

Science Daily
October 29, 2013

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Badgers ultimately responsible for around half of TB in cattle

Badgers are ultimately responsible for roughly half of tuberculosis (TB) in cattle in areas with high TB prevalence, according to new estimates based on data from a previous badger culling trial. However, only around six per cent of infected cattle catch TB from badgers, with onward transmission between cattle herds accounting for the remainder, the study suggests. The findings are published in the journal PLOS Currents: Outbreaks. The role of badgers in spreading bovine TB has been debated intensely as part of discussions about whether badgers should be culled to control the disease. The Randomised Badger Culling Trial, which ran from 1998 to 2005, found evidence that culling could reduce TB in herds inside culled areas, while increasing TB in areas nearby. Mathematical models based on data from the trial were previously used to calculate an estimate of the proportion of TB in cattle that could ultimately be attributed to transmission from badgers. The new paper, by scientists at Imperial College London, provides a more detailed analysis. It estimates that badgers ultimately account for 52 per cent of cattle TB in areas where prevalence in cattle is high. There is considerable uncertainty around this estimate, but the authors say that 38 per cent is a robust minimum value for the estimate. Professor Christl Donnelly, from the Medical Research Council Centre for Outbreak Analysis and Modelling at Imperial, said: “These findings confirm that badgers do play a large role in the spread of bovine TB. These figures should inform the debate, even if they don’t point to a single way forward.” The mathematical model suggested that 5.7 per cent (the 95% confidence interval for this estimate is 0.9% to 25%). of transmission to cattle herds is from badgers to cattle, with the rest of the contribution from badgers resulting from onward transmission between cattle herds.

Science Daily
October 29, 2013

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Alternative to antibiotics: Plasmas attack bacterial cells on several levels

As they destroy bacteria very efficiently, plasmas constitute an alternative to chemical disinfectants and potentially to antibiotics, as well. How they achieve this effect has been investigated by biologists, plasma physicists and chemists at the Ruhr-Universität (RUB). Cold atmospheric-pressure plasmas attack the prokaryote’s cell envelope, proteins and DNA. “This is too great a challenge for the repair mechanisms and the stress response systems of bacteria,” says Junior Professor Dr Julia Bandow, Head of the Junior Research Group Microbial Antibiotic Research at the RUB. “In order to develop plasmas for specific applications, for example for treating chronic wounds or for root canal disinfection, it is important to understand how they affect cells. Thus, undesirable side effects may be avoided right from the start.” The team reports in the Journal of the Royal Society Interface. Depending on their specific composition, plasmas may contain different components, for example ions, radicals or light in the ultraviolet spectrum, so-called UV photons. Until now, scientists have not understood which components of the complex mixture contribute to which extent to the antibacterial effect. Julia Bandow’s team has analysed the effect of UV photons and reactive particles, namely radicals and ozone, on both the cellular level and on the level of single biomolecules, namely DNA and proteins. On the cellular level, the reactive particles alone were most effective: they destroyed the cell envelope. On the molecular level, both plasma components were effective. Both UV radiation and reactive particles damaged the DNA; in addition, the reactive particles inactivated proteins. Atmospheric-pressure plasmas are already being used as surgical tools, for example in nasal and intestinal polyp extraction. Their properties as disinfectants may also be of interest with regard to medical applications. “In ten years, bacteria might have developed resistance against all antibiotics that are available to us today,” says Julia Bandow. Without antibiotics, surgery would become impossible due to high infection rates.

Science Daily
October 15, 2013

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Bacteria from lean cage-mates help mice stay slim

Gut bacteria from lean mice can invade the guts of obesity-prone cage-mates and help their new hosts to fight weight gain. Researchers led by Jeffrey Gordon, a biologist at Washington University in St. Louis, Missouri, set out to find direct evidence that gut bacteria have a role in obesity. The team took gut bacteria from four sets of human twins in which one of each pair was lean and one was obese, and introduced the microbes into mice bred to be germ-free. Mice given bacteria from a lean twin stayed slim, whereas those given bacteria from an obese twin quickly gained weight, even though all the mice ate about the same amount of food. The team wondered whether the gut microbiota of either group of mice would be influenced by mice with one type living in close quarters with animals harbouring the other type. So the scientists took mice with the ‘lean’ microbiota and placed them in a cage with mice with the ‘obese’ type before those mice had a chance to start putting on weight. “We knew the mice would readily exchange their microbes,” Gordon says — that is, eat each other’s faeces. Sure enough, the populations of bacteria in the obese-type mice changed to match those of their lean cage-mates, and their bodies remained lean, the team writes in Science.

The bacterial invasion travelled only in that direction, however: the bacteria of the obese mice could not colonize the lean neighbour. This makes sense, says Gordon, who found in earlier work that the population of gut bacteria in obese people is less diverse than that in lean people, leaving unfilled niches in the microbiota. The bacteria from the lean mice seem to be able to find those vacancies, he says. But this left him wondering: if the bacteria of lean people are so good at setting up shop in the guts of the obese, “why don’t we have an epidemic of leanness in America?” So the team fed the mice a more human diet, turning foods such as breakfast cereal and pizza into pellets for the mice. When the animals were fed a diet low in saturated fat and high in fruit and vegetables, the transfer of gut microbes from mice with the lean type to those with the obese type still occurred; however, when the mice were given a high-fat, low-vegetable diet this did not happen, and mice with the obese-type bacteria gained weight. “There’s an intricate relationship between our diet and how our gut bugs work,” says Gordon. “You have to have the right ingredients.” Patrice Cani, who studies the interaction between gut bacteria and metabolism at the Catholic University of Leuven in Belgium, is impressed that the authors of the study were able to demonstrate causality between gut microbiota and a physical feature such as body type. And he says there is much more to be learned about the interaction of gut bacteria and diet from the work. “This paper is like a bank of information,” he says. “We can keep going back for a deeper look.”

Nature
October 1, 2013

Original web page at Nature

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Copper destroys highly infectious norovirus

Scientists from the University of Southampton have discovered that copper and copper alloys rapidly destroy norovirus — the highly-infectious sickness bug. Worldwide, norovirus is responsible for more than 267 million cases of acute gastroenteritis every year. In the UK, norovirus costs the National Health Service at least £100 million per year, in times of high incidence, and up to 3,000 people admitted to hospital per year in England. The virus, for which there is no specific treatment or vaccine, can be contracted from contaminated food or water, person-to-person contact, and contact with contaminated surfaces, meaning surfaces made from copper could effectively shut down one avenue of infection. The study, which was designed to simulate fingertip-touch contamination of surfaces, showed norovirus was rapidly destroyed on copper and its alloys, with those containing more than 60 per cent copper proving particularly effective. Copper alloys have previously been shown to be effective antimicrobial surfaces against a range of bacteria and fungi. The Southampton research reported rapid inactivation of murine norovirus on alloys, containing over 60 per cent copper, at room temperature but no reduction of infectivity on stainless steel dry surfaces in simulated wet fomite and dry touch contamination. The rate of inactivation was initially very rapid and proportional to the copper content of alloy tested. Viral inactivation was not as rapid on brass as previously observed for bacteria but copper-nickel alloy was very effective.

One of the targets of copper toxicity was the viral genome and a reduced number of the gene for a viral encoded protein, VPg (viral-protein-genome-linked), which is essential for infectivity, was observed following contact with copper and brass dry surfaces. Lead author Sarah Warnes, from the Centre for Biological Sciences at the University of Southampton, says: “The use of antimicrobial surfaces containing copper in clinical and community environments, such as cruise ships and care facilities, could help to reduce the spread of this highly infectious and costly pathogen. “Copper alloys, although they provide a constant killing surface, should always be used in conjunction with regular and efficient cleaning and decontamination regimes using non-chelating reagents that could inhibit the copper ion activity.” Co-author Professor Bill Keevil, from the University’s Institute for Life Sciences, adds: “Although the virus was identified over 40 years ago, the lack of methods to assess infectivity has hampered the study of the human pathogen. “The virus can remain infectious on solid surfaces and is also resistant to many cleaning solutions. That means it can spread to people who touch these surfaces, causing further infections and maintaining the cycle of infection. Copper surfaces, like door handles and taps, can disrupt the cycle and lower the risk of outbreaks.” The study ‘Inactivation of norovirus on dry copper alloy surfaces’ is published in the latest issue of the journal PLOS ONE. Previous laboratory studies by the University of Southampton have described the rapid death of bacterial, fungal and viral pathogens such as MRSA on copper alloy surfaces and also prevention of antibiotic resistance horizontal gene transfer between pathogens.

Science Daily
October 1, 2013

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Vaccinating cattle against the E. coli O157 bacterium could cut the number of human cases of the disease by 85%, according to scientists

The bacteria, which cause severe gastrointestinal illness and even death in humans, are spread by consuming contaminated food and water, or by contact with livestock feces in the environment. Cattle are the main reservoir for the bacterium. The vaccines that are available for cattle are rarely used, but could be significant. The research was lead by a team of researchers at the University of Glasgow in collaboration with the University of Edinburgh, the Royal Veterinary College, Scotland’s Rural College, Health Protection Scotland, and the Scottish E. coli O157/VTEC Reference Laboratory. The study, published in the online journal PNAS, used veterinary, human and molecular data to examine the risks of E. coli O157 transmission from cattle to humans, and to estimate the impact of vaccinating cattle. The risk of E. coli O157 infection is particularly significant when the cattle are ‘super-shedding’ — excreting extremely high numbers of bacteria in their feces for a limited period of time. Vaccines against the bacteria exist that can reduce super-shedding. As a consequence, the researchers predict that vaccinating cattle could reduce human cases by nearly 85 percent, far higher than the 50 percent predicted by studies simply looking at the efficacy of current vaccines in cattle.

These figures provide strong support for the adoption of vaccines by the livestock industry, and work is now underway to establish the economic basis for such a program of vaccination. In addition, research is continuing in Scotland by the same collaborative grouping to develop even more effective vaccines that would further reduce the impact on human disease. Lead author, Dr Louise Matthews, Senior Research Fellow in the Institute of Biodiversity, Animal Health and Comparative Medicine, said: “E. coli O157 is a serious gastrointestinal illness. The economic impact is also serious — for instance studies in the US suggest that healthcare, lost productivity and food product recalls due to E. coli O157 can cost hundreds of millions of dollars each year. “Treating cattle in order to reduce the number of human cases certainly makes sense from a human health perspective and, while more work is needed to calculate the cost of a vaccination program, the public health justification must be taken seriously.” In Scotland, an average of 235 culture positive cases of E. coli O157 infection per year (i.e. people who had the organism in their stools) were notified to Health Protection Scotland from 2008 to 2012. The vaccines that are available currently have poor take-up: one version in the US is not fully licensed because medicines for veterinary use must show that animal health is improved. This is problematic because E. coli O157 does not harm cattle and assessing the impact of treatment involves coordination between human and veterinary health practitioners.

Science Daily
October 1, 2013

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Food-borne illnesses are not always home-grown

Scotland’s highland cows have been badmouthed as promoters of salmonella outbreaks, but the bacteria may have come from abroad instead. Scottish cows have a bum rap. For decades, the local cattle have been prime suspects behind the country’s outbreaks of drug-resistant, food-borne illnesses. But research now suggests that humans and imported foods are the real culprits. A team of researchers compared the genome sequences of nearly 400 samples of diarrhoea-causing Salmonella enterica collected from people and livestock in Scotland. They found that bacterial strains infecting humans were largely distinct from those found in local cattle, but had close ties to strains that had been isolated in other countries. The results suggest that mass epidemics may spark from a complicated intermingling of bacteria between animals and humans and from exchanges between different countries, the authors say. Their findings are published in Science. “There is a pervading wisdom that local animals are a predominant source of pathogens and resistance,” says study co-author Stuart Reid, a veterinary epidemiologist at the Royal Veterinary College in Hatfield, UK. But as his team’s findings show, that may not always be the case. “It’s only if we can treat this as an international issue that we’re going to get to the bottom of it,” he says.

Reid and his colleagues focused on Scottish outbreaks because of the country’s ample collection of bacterial samples obtained from both humans and livestock. The collection was started 23 years ago, when global epidemics of drug-resistant salmonella infections began to arise. Livestock was assumed to be the source of the epidemics because animals naturally harbour the bacteria. To find out whether this was really the case, the team used whole-genome sequencing to trace the tiny evolutionary steps of the collected bacterial strains. They analysed 142 samples isolated from Scottish patients and 120 from local animals, mostly cows, then compared them with 111 strains collected from people and animals in other countries. The team found that strains infecting Scottish patients were different from those in local livestock. And they noted only a few instances in which strains isolated from local livestock had spread to humans. But they also found that strains could spread from humans to animals. “It’s occurring at a low frequency, but in both directions,” explains lead author Alison Mather, an epidemiologist at the Wellcome Trust Sanger Institute near Cambridge, UK.

When they looked at the strains’ antimicrobial resistance, the researchers found that bacteria from humans had more diverse collections of resistance genes than those in local livestock. This indicates that local livestock cannot be the sole source of the resistance genes found in the strains found in humans. The authors therefore suggest that local livestock are not the source of drug-resistant human salmonella outbreaks in Scotland. Rather, they say, foreign strains carried by other humans and in imported food probably entered the country and infected animals and humans separately, then continued to evolve and acquire resistance separately. The authors stress that the study does not imply that antimicrobial resistance developed on farms is less concerning than previously thought, including resistance stemming from the controversial practice of giving antibiotics in feed to promote animal growth. “We’re not saying it’s not as bad, we’re just saying that there are other sources that need to be considered,” says Reid. Though local animals were not a main source of these pathogens, he explains that it does not eliminate the possibility that resistance genes from local farms and foreign farms played a role. Mark Woolhouse, an epidemiologist at the University of Edinburgh, UK, says that the study clarifies how pathogens and drug-resistance genes spread. “It’s not just multi-bug, multi-drug,” he says, “but multi-country.”

Scotland imports most of its red meat, but the authors say that the country does not have adequate surveillance in place to determine whether imported food is a source of new pathogens. Both Woolhouse and the authors call for Scotland and other countries to boost the monitoring of their food supply. Lance Price, a genomic epidemiologist at the George Washington University in Washington DC, says that it is not surprising that Scottish cattle are not the source of Scottish outbreaks, because the epidemics were international. He notes, however, that, to eliminate the possibility of a domestically derived outbreak, the authors should have analysed more strains from poultry and pigs, which also carry S. enterica. “But it underscores that this is a global issue,” he says. “Meat sale and meat trade across borders is making it harder to control antibiotic-resistant pathogens at a local scale.” Mark Woolhouse, an epidemiologist at the University of Edinburgh, UK, says that the study clarifies how pathogens and drug-resistance genes spread. “It’s not just multi-bug, multi-drug,” he says, “but multi-country.” Scotland imports most of its red meat, but the authors say that the country does not have adequate surveillance in place to determine whether imported food is a source of new pathogens. Both Woolhouse and the authors call for Scotland and other countries to boost the monitoring of their food supply. Lance Price, a genomic epidemiologist at the George Washington University in Washington DC, says that it is not surprising that Scottish cattle are not the source of Scottish outbreaks, because the epidemics were international. He notes, however, that, to eliminate the possibility of a domestically derived outbreak, the authors should have analysed more strains from poultry and pigs, which also carry S. enterica. “But it underscores that this is a global issue,” he says. “Meat sale and meat trade across borders is making it harder to control antibiotic-resistant pathogens at a local scale.”

Nature
October 1, 2013

Original web page at Nature