Meet the Microbiologist

How can the humble zebrafish teach us about the human microbiome? John Rawls discusses the benefits of using animal models Take the MTM Listener Survey  Julie’s Biggest Takeaways:   Zebrafish and other model animals provide opportunities to understand host-microbe interactions. Zebrafish are particularly useful for imaging studies, due to their translucent skin and the ease of in vivo microscopy. This allows zebrafish to be used to in studies of spatial architecture or longitudinal studies (imaging the same fish specimen over time) in ways that other model organisms can’t be.   Zebrafish get their first microbes from their mother, just like mammals! The chorion, a protective coating that surrounds the zebrafish embryo, is seeded with microbes from passing through the cloaca of the female zebrafish. Surface-sterilizing this chorion allows researchers to generate germ-free animals that are very useful for microbiome studies.   A gut epithelial transcription factor is regulated by a signal from the gut microbiota, and this signaling interaction is conserved among all vertebrates. The transcription factor itself, HNF4, is found in both complex and simple animals, like the sea sponge, and may serve a long-conserved function in regulating interactions between animals and their microbiota.   Enteroendocrine cells release hormones based on specific chemical cues, but they can also interact with the nervous system. This makes them an important part of the gut-brain system, and the power of in vivo imaging has made zebrafish a great model for better understanding their function. Specific members of the microbiome specifically stimulate these EECs, sending signals up the vagus nerve to the brain.   Featured Quotes:   “We know that the zebrafish functionality of its intestine is very similar to what one encounters in the mouse or human intestine and we and others have been able to translate our findings from zebrafish studies into human biology.”   On genomic studies that have found similar transcription profiles in zebrafish, stickleback fish, mice, and humans: “This suggested that there is a core transcriptome that gut epithelial cell use in different vertebrate species that haven’t shared an ancestor in 420 million years!”   Comparing fish and mouse: “Genes regulated by microbiota in these respective hosts display a lot of overlap. Many of the same signaling pathways and metabolic processes are affected by microbiotas in different hosts in similar ways.”   “There’s been a lot of interesting research documenting the role of the intestinal microbiome in promoting harvest of dietary nutrients we consume. Much of that literature has been focused on the events that occur in the distal intestine, in the colon, where recalcitrant carbohydrates and proteins that make it that far, many of which we are unable to digest, are made available to the colonic microbiome, members of which are able to digest and degrade them to things such as short chain fatty acids, which we can consume.”   “Eventually, we’ll have some strong candidates in terms of specific bacterial strains or communities or factors or pharmacologic agents that could be used to affect dietary fat absorption or metabolism. We’re still a long ways away from that.”   “One of the fascinating things about developmental biology is that the only way you get a viable animal is if the different tissues and the different cells within the body are coordinating amongst themselves for energy, for nutrients, for oxygen, et cetera. As you’re building an animal and as you’re sustaining an animal, the different tissues have to cooperate. When that doesn’t happen, when tissues or cells become selfish or don’t play by the rules, you get things like cancer and other diseases as well...when I began learning about the field of microbiome science and some of the work that was coming out from that field, it sounded to me like the microbiome was going to be a really important part of that. Not only can we think of the microbiome as a ‘microbial organ,’ as it is sometimes called, and therefore worthy of consideration within the context of developmental biology, but also the influence of the microbiome on any one tissue is going to modify its need and its ability to cooperate within the integrated system.”   Links for this Episode:   John Rawls’ lab website More amazing zebrafish images from the Rawls lab Duke University Microbiome Center Genome Research article on HNF4 regulation Cell Host and Microbe article on microbial influence on fatty acid absorption  

Gemma Reguera discusses her studies of Geobacter pili, which transfers electrons to iron oxide and other minerals, and can be used for new biotech applications. Host: Julie Wolf  Subscribe (free) on Apple Podcasts, Google Podcasts, Android, RSS, or by email. Also available on the ASM Podcast Network app. Julie’s Biggest Takeaways: Geobacter sulferreducans, a bacterium that “breathes” rust, is the lab representative of the genus Geobacter that dump electrons onto rust. These specialized microbes use minerals like manganese oxide and iron oxide (also known as rust) for respiration in both terrestrial and aquatic sediments. Although many species are strict anaerobes, a few species can grow under microaerophilic conditions, in which the bacteria will respire the oxygen to eliminate its toxic effects on the cell. Iron oxide respiration relies on the Geobacter pili, a simple structure composed of a single peptide repeat. The pili concentrate on one side of the bacterial cell, where they connect the cell with the iron oxide to release the electrons that have been accumulating. The pili immediately depolymerize and retract, shedding the mineral before returning into the cell. Mass-producing pilin subunits in E. coli took a bit of trouble shooting, but now Reguera and her colleagues can make them on a much larger scale, which bodes well for expanding tests into electronic applications. Commercialization grants address the “valley of death,” the chasm between the technologies developed at the bench and the scale of production necessary for industrialization. Geobacter can bind and reduce many minerals using their pili, including uranium and other toxic heavy metals like lead and cobalt. Using Geobacter pili in agricultural soils or aquaculture waters may help remove these contaminants and improve the health of these ecosystems. Featured Quotes: “I remember when I started as a microbiology student, I think I underappreciated the role that electrons and the movement of electrons play in microbiology.” “There is absolutely not a single process in living organisms that is not energized by the movement of electrons.” “The Earth didn’t have oxygen for the first 2 billion years, if not longer - and there was life! On Earth! Those early organisms were really great at finding minerals, metals, just about anything other than oxygen to dump their electrons, continue to grow, and to colonize the Earth.” “When you start comparing the structure and the amino acid composition of this subunit to any other known bacterial pilins, you really see 2 remarkable changes: one of them is the pilin of Geobacter is very small. the second is that little stick has aromatic amino acids. When the sticks come together to make the filament, they cluster very close to each other and create like a staircase for the electrons to move fast. It’s like a magic combination in which you have the right structural reduction and the right amino acids to really fit like a puzzle to create paths for electrons.” “What has always motivated me is learning something new.” Links for This Episode: Gemma Reguera lab website Gemma Reguera interview on “People Behind the Science” HOM: Thirty-Second Annual Meeting of the Society of American Bacteriologists HOM: Barney Cohen: An Appreciation (Bacteriological Reviews memorial)

Why do Legionnaire’s Disease outbreaks occur mostly in the summer? What is the connection of the Flint change in water source and Legionella outbreaks in the area? Michele Swanson discusses her work on Legionella pneumophila and her path from busy undergraduate to ASM President. Julie’s Biggest Takeaways: Legionella pneumophila is a waterborne microbe that lives in fresh water and can colonize water systems of the built environment. Colonization of cooling systems, like those used in air conditioning systems, can lead to contaminated water droplets that can cause disease. Legionella are very adaptable to different environment, but scientists don’t have great models to determine the exact preferences of the bacterium. After Flint switched water sources from lake to the Flint river, a chemical that prevents corrosion was omitted from the water treatment. This led to lead in the water, which was detected in pediatric patients. An increase of legionella cases in the two years also occurred, and the question was whether the outbreak was related to the shift in water chemistry. Michele joined a team of water engineers, epidemiologists and sociologists to answer this question, and the team found an association between low chlorine levels and high risk of legionella disease. Across the globe, more than 80% of disease is associated with L. pneumophila serogroup 1. The serogroup is based on the bacterial lipopolysaccharide (LPS) structure, which in this strain is very hydrophobic and may allow this serogroup to withstand a higher degree of desiccation than other strains. A urine-based diagnostic test works well, but only to detect serogroup 1. The strain isolated from patients of the Flint outbreak were serogroup 6, as were Legionella isolated from the homes of Flint residents. Featured Quotes: “Amoeba are very good at digesting bacteria, eating them for food, but Legionella, because it’s been under this severe selective pressure of the amoeba, they’ve evolved tools to allow them not only to survive within the amoeba but to replicate within the vacuole of the amoeba.” “We now have equipment that throws water into the air and gives [Legionella] a new opportunity to be ingested by a macrophage. It can then deploy the same tricks it uses to grow inside amoeba to grow inside the macrophage.” “[Human infection] is a tragedy for the patient, but also for the microbe...humans are a dead-end for the bacterium.” “I was really delighted to be recruited to work with this interdisciplinary team on a public health crisis here in my home state. It has opened my eyes to a much more complex pathway and I just feel really privileged in this stage in my career to be able to turn my attention to these larger public health issues.” “People want to hear encouragement; we have a tendency to compare ourselves to those who are 5-10 years ahead of us. Encouragement really is valuable.” Links for this Episode: Michele Swanson at the University of Michigan mBio: Prevalence of infection-competent Legionella pneumophila within premise plumbing within southeast Michigan PNAS: Assessment of the Legionnaire’s disease outbreak in Flint, Michigan Microbial Sciences blog post: Examining Flint: New research highlights lack of Legionella public policy ASM membership  

How is Toxoplasma gondii, a protozoan that causes neuro-invasive disease, transmitted as a foodborne pathogen? Why are cats important in transmitting Toxoplasma infection? Anita Koshy answer these questions and talks about her research on the latest Meet the Microbiologist. Julie’s Biggest Takeaways: The primary host for T. gondii is cats, in which the protozoan can undergo sexual reproduction. Why cats? No one knows, in part because there isn’t a good in vitro system to study cat epithelial cell interactions with T. gondii. Most warm-blooded animals, including birds, can be infected with Toxoplasma. Intermediate hosts can pass Toxoplasma from one to another if one eat these tissue cysts, explaining why Toxoplasma can be a foodborne pathogen. In healthy individuals, the immune response clears most fast-growing cells (tachyzoites) but some protozoans convert to a slow-growing cell form (bradyzoites). In people, these bradyzoites form cysts predominantly in the brain, the heart and the skeletal muscle. Some serological studies suggest a tie between Toxoplasma infection and brain disorders, but these are less definitive than causative studies in mice. Populations with high Toxoplasma or low Toxoplasma prevalence don’t see a correlative incidence of disorders such as schizophrenia or Alzheimer’s disease. Featured Quotes: “When we talk about neuroinfectious diseases, we’re talking about the diseases that cause symptoms. Those that can get into the central nervous system and those that cause symptomatic disease are the same.” “A parasite is sitting there dormant or maybe reactivating every so often and the immune system comes in and deals with that reactivation. But when you lack an immune system, all of a sudden when that parasite reactivates, there is no longer this immune system that will come in and clear it out.” “What we don’t know is whether reactivation occurs preferentially in the brain. There is evidence from HIV patients of inflammation of the heart or inflammation of the skeletal tissue - but those weren’t the symptoms that presented, which were of the brain. Did reactivation happen in the brain, or did it occur elsewhere and the parasite was able to travel to the brain and there’s no longer an immune system to clear it out?” Links for this Episode: Koshy Lab Site Sea Otter Infection with Toxoplasma Rats Infected with T. gondii Lose Their Aversion to Cat Urine HOM Tidbit: The History of Toxoplasma gondii Bill Hutchinson obituary  

Jennifer Gardy talks about whole-genome sequencing as a technique to address public health issues using genomic epidemiology. She talks about her research on TB and new DNA sequencing technologies, including her vision for microbial genetic sequencing as one piece of the puzzle in the future of public health. Julie’s Biggest Takeaways: Whole-genome sequencing technologies are replacing older DNA technologies to identify relatedness between microbial isolates. The genome sequences help to identify epidemiological questions such as the origins of an outbreak. A pathogen’s genome being passed person-to-person accrues small changes, similar to children playing telephone - except those children are scattered around the room, and you have to logically deduce the order in which the information was passed. DNA sequencing has moved forward faster than the upstream genomic preparation and downstream sequence analysis areas; Gardy expects advances in these ‘bookend’ areas to be breakthroughs of the future. The Ebola and Zika outbreaks were test cases for portable DNA sequencing technologies, but informative based on the different disease presentation: Ebola patients have high viral loads and thus a lot of genomic material, but Zika patients have much lower viral loads and it was much harder to get samples. Based on pathogen characteristics, DNA sequencing can identify the end of an outbreak. Gardy used sequencing to find that patients with TB, which can take years to develop into fulminant disease, had been infected years previous, and was able to see that transmission was no longer ongoing. Featured Quotes: “Genomics is really cool because instead of interviewing people about what happened in an outbreak, we’re interviewing the pathogen!” “[Working at BCCDC] is a really nice ecosystem, where you can really see the results of your research changing public care policy and practice in real time, and that is incredibly rewarding.” “The only prediction you can make about DNA sequencing is there’s always going to be something new and different.” “Depending on your use-case, sometimes you need to go after the whole genome and other times a targeted approach is more than enough.” “I’m excited to see how this [microbial DNA sequencing] work fits in into an overall public health landscape. It’s cool to sequence genomes and make some reports about transmission networks, but that’s just one small part of a very big public health system that is trying to keep populations healthy. It requires so many different people, from nurses and doctors on the frontline to policy makers behind the scenes to social scientists who are interacting with patients or care providers to people that are understanding the economics of these things... when you start to see how these different pieces of the puzzle fit together, I think there’s a lot of opportunities in the future for making microbial genomics just one piece of a large interdisciplinary puzzle of people that are working together across different fields to address a disease from multiple different angles.” Links for This Episode: Jennifer Gardy’s website Jennifer Gardy at UBC Nanopore Minion Alan Alda Center for Communicating Science Banff Science Communications HOM Tidbit: Albrecht Kossel, a Biographical Sketch

See the full shownotes at: asm.org/mtm Julie Pfeiffer tells the story of how she serendipitously found a role for the gut microbiota during polio virus infection, and how she and her lab discovered an important role for bacterial glycans in viral biology. She also talks about viral fitness strategies, and how RNA viruses and DNA viruses benefit from making different amounts of errors when copying their genomes. Julie's biggest takeaways: Determining the exact nature of the glycans that play these roles has been difficult because they are very complex. Aspects of lipopolysaccharide, chitin, and peptidoglycan are all sufficient to bind the viral capsid, but because of their structural complexity, it’s difficult to pinpoint the exact molecular interaction. Bacterial glycan interactions with viruses benefit the virus in two ways: the virus can be delivered to a host cell it will infect, and the viral capsid is stabilized. Whether there is a benefit to the bacterium during these interactions is unknown, but is an active area of research in Julie’s lab. Many viruses can be inactivated at body temperature or even room temperature if they prematurely release their genetic material. Polio viruses are simply a protein shell surrounding an RNA genome, and the capsid can ‘breathe,’ slightly changing its conformation. Sometimes, the genome is accidentally released, resulting in a viral dead end. Julie showed that bacterial glycans will lock the capsids into a conformation and prevent genome release from happening until the virus encounters a host cell. Julie is a proponent of clear communication, including with those working in similar fields, which she learned from her experience as a postdoctoral fellow. She and a postdoc in a different institution, Marco Vignuzzi, independently isolated a polio virus mutant that made fewer in genome replication. Both showed that the virus had a defect during mouse infection, indicating that the ability to introduce errors during genome replication is beneficial to viral fitness. Julie and Marco finally met at a viral evolution conference, after which they became close friends. Featured Quotes (in order of appearance): “I get more excited about a surprising result because it probably means there’s some interesting underlying biology that couldn’t be anticipated!” “We’ve done many gross experiments, so buyer beware; you’ve got to know what you’re getting into [with a fecal-oral pathogen].” “The infectious unit may be more complicated than we think!” “Communicating with people you know working on similar things can be mutually beneficial for everyone: you both get credit; nobody gets scooped. It’s win-win for sure.” “The truth is most enteric viral infections are self limiting in most healthy individuals so you’re much better off trudging through a day or two of gastrointestinal illness than blowing up your microbiota.” Links for this episode Julie Pfeiffer website at UT Southwestern Medical Center Back-to-back Science publications from Golovkina and Pfeiffer PLOS Pathogens: The importance of model systems: Why we study a virus on the brink of global eradication Viruses and Cells Gordon conference (donate here) HOM Tidbit: Michael Underwood’s A Treatise on the Diseases of Children

Pete Greenberg tells how bacteria can communicate based on cell density, a phenomenon he helped name quorum sensing. He talks about therapeutics based on quorum-sensing discoveries, and how studying bacterial interactions can be used to test ecological principles like cooperation and social cheating. Julie's biggest takeaways: Quorum sensing can be likened to an old-fashioned smoking room, where a few cigar smokers don’t affect the air quality, but as more smokers enter the room, it becomes beneficial to the group to open the window: a changed behavior that benefits the group environment. Differentiating waste molecules from signaling molecules is important to define specific quorum sensing. The experimental evidence that shows that molecules serve as quorum sensing signals that allow bacteria to respond at high density comes from social engineering experiments to identify ‘cheaters.’ Quorum sensing results in changes in gene expression that benefit the community but not necessarily individual cells. An example is antibiotics, which when made by a single cell aren’t at a high enough concentration to kill competitor microbes. As a group, all cells working together can produce a cloud of antibiotic that may be able to protect from competitors. The ability of microbes to receive or ‘eavesdrop’ on the signals produced may be cooperative, but is more likely competitive, giving the eavesdropper a competitive advantage by informing them about another species’ presence. If you knock out quorum sensing, you get abnormal biofilms, but it doesn’t ablate biofilms completely. Although a self-described disinterested high-school student, Greenberg signed up for a weekend field trip to get out of a test on a Friday. It was looking at animals in the intertidal bay of the Pacific Northwest that inspired him to be a biologist! Greenberg also credits his broad biology undergraduate training for preparing him to apply socioecology concepts to bacteria. Quorum sensing was originally called ‘auto induction.’ In the early 1990s, Greenberg was writing a minireview for the Journal of Bacteriology and wanted to think of a catchy title. As Greenberg remembers, coauthor Steve Winans explained the concept to his family, and his brother-in-law said “it’s like the bacteria need a quorum” - the birth of the term ‘quorum sensing.’ Featured Quotes (in order of appearance): “So-called ‘cheaters’ don’t respond to the signal, they’ve lost the ability to respond to the signal. The product that’s useful for the common good any more. They don’t pay the cost of cooperation but they can benefit by the cooperative activity of everyone else in the community...there’s a fitness advantage for cheaters in this environment.” “It’s a real case of convergent evolution. It’s important that the bacteria can do this, and these two really distinct types of [gram-positive and gram-negative] bacteria have evolved completely different mechanisms to perform quorum sensing.” “I think of bacteria as a way to study what is called ‘Darwin’s dilemma.’ If a cheater emerges among a population, it will have a fitness advantage over the population of cooperators. It should take over the population and ultimately cause the tragedy of the commons, where there are too many cheaters and not enough cooperators and the whole system collapses. Darwin’s dilemma is: how is cooperation stabilized? We know it exists and it seems like it shouldn’t - we can use bacteria to get at the rules.” “I got interested in [quorum sensing] because it was so cool!” “I had this idea, as we began to unravel quorum sensing in these marine luminescent bacteria, that any idea in biology that’s a good idea will occur more than once - but I didn’t have any evidence of that. For 15 years, my lab and essentially one other lab, Mike Silverman’s lab, were the only labs working on this. It was really the early 90s when our group and other groups started to realie that lots of bacteria do this. It’s one of those fantastic oddesies. It’s luck - luck and hard work, I guess. Hard work by the people in my lab as I sit around as watch!” “It’s funny how a term can catch on and sort of crystallize a field! But somehow, it seemed to do that. I’ve gotten really into trying to think of catchy terms since then, and the latest one is ‘sociomicrobiology,’ which I introduced with Matt Parsek about 12 years ago and there’s a burgeoning field called sociomicrobiology. I’m trying to think of another term now, before I retire!” Links for this episode   Pete Greenberg lab at the University of Washington Pete Greenberg 2004 PNAS bio Journal of Bacteriology minireview: Quorum Sensing in Bacteria HOM: Woody Hastings memoriam ASM Podcasts     Send your stories about our guests and/or your comments to jwolf@asmusa.org.

Charlie Rice gives the history of learning to grow hepatitis C virus in culture, from pitfalls to hurdles and successes along the 20-year journey. He also talks about yellow fever virus, its vaccine, and the importance of curiosity-driven research

How are new diseases detected in a clinical microbiology lab? Melissa Miller talks about the time it takes to develop a test for a new disease (hint: it’s getting shorter). She also shares her definition of ‘point-of-care’ diagnostics and explains the major trends for clinical microbiology labs. Host: Julie Wolf  Subscribe (free) on iPhone, Android, RSS, or by email. You can also listen on your mobile device with the ASM Podcast app. Julie's biggest takeaways: Antibiograms are vital to understand the resistance characteristics of locally circulating disease strains. These help make empirical decisions for antibiotic therapy regimens before the susceptibility test results are available. New diseases require new diagnostic tests. How to determine how well new tests work once they’re developed? Clinical microbiologists look to the sensitivity (how well does a test detect if a patient has a disease) and specificity (how often is the test negative if the patients doesn’t have it) of the test. Having access to positive controls (that is, samples from a patient known to have the disease) can prove difficult in some settings, such as in North Carolina, where no Zika patients were admitted while the Zika virus test was being developed. When the HIV epidemic was beginning, it took several years after the HIV virus was identified to sequence its genome and use this for molecular testing. In 2002-2003, it took just over a month to get the SARS genome sequence for use in developing assays. It’s even quicker today; within a week, we can have sequences from viruses around the world. Defining ‘point-of-care’ testing took an entire hour at a recent American Academy of Microbiology colloquium! Melissa’s take: It’s a test that can be done at or near to where the patient is. Point-of-care tests are exciting but can also pose challenges. A recent example is false-positive pertussis tests that were shown to be due to pertussis vaccine being administered nearby. Ensuring the tests are used safely and accurately will best serve healthcare workers and patients alike. Molecular diagnostics have two trends: one trend simplifies existing technologies into point-of-care tests. The other trend adds complexity, by applying next-generation sequencing techniques in a reproducible manner. Featured Quotes (in order of appearance): “Laboratorians are often in the basement or in a setting where they aren’t visible to the healthcare team, but they’re very vital to taking care of the patient.”   “When you’re using laboratory-developed tests, the way it works in one laboratory may be very different from how it works in another laboratory.”   “The ultimate goal [of point-of-care testing] is to get a result that’s actionable. We don’t need to do tests that aren’t going to result in a clinically actionable decision.”   “In many ways, the technology is ahead of where our quality assurance protocols are.”   “I think it’s going to be very important in going ahead that we continue to have laboratorians involved in developing these point-of-care programs and consulting to these sites, helping to make sure that there are policies and procedures that ensure quality results for their patients.”   “It’s one thing to do it in a research setting; we’ve collaborated with a number of folks using next generation sequencing. But to then move it to the clinical lab and have it be reproducible and have the quality at the level you need for a clinical lab is a completely different challenge.” Links for this episode Melissa Miller University of North Carolina Website Division of Clinical Laboratory Science at University of North Carolina Searchable List of Clinical Laboratory Science Programs AAM Colloquium Report on Point-of-Care Testing CPEP Program Career Blog: Tips on becoming a clinical microbiology laboratory director HOM Tidbid: Papagrigorakis 2006 International Journal of Infectious Diseases report HOM Tidbit: Shapiro reply to Papagrigorakis report Send your stories about our guests and/or your comments to jwolf@asmusa.org.  

Veterinarian and epidemiologist Mathew Muturi tells how a Rift Valley Fever outbreak led to implementation of One Health-based policies. Muturi talks about his One Health training and its applications for health and biopreparedness. Julie’s Biggest Takeaways: One Health Simple communication between experts helps facilitate implementation of one health in public systems. Sitting experts in human and animal health in the same office allows easier communication between these different health sectors. One Health policies involving close collaboration between animal and human healthcare workers were first implemented in Kenya in response to the threat of avian influenza, but were discontinued after the threat waned. Human cases of Rift Valley Fever, due to spillover from a livestock outbreak, led to the discovery that these collaborative policies could prevent other outbreaks as well, and the policies were reinstated. Zoonotic diseases can often be the most overlooked. Officials of countries where endemic diseases are present may have preparedness plans for serious cases but may overlook something endemic like brucellosis. There are 42 subtribes in Kenya, including diverse languages, religions, and beliefs. Public health interventions do their best to align the local beliefs of the people to minimize risk of pathogen exposure. Featured Quotes: “One health is not a new concept; it’s an old concept that explains the health of humans, animals, and the environment is interconnected. It’s a concept that plays out in everyday life.” “One of the reasons One Health has been able to be successful in Kenya, and that I suggest to other countries wishing to implement this program, is the sitting together, talking together. Make sure that you work together, see each other - I don’t think communication works well enough if it’s on an ad hoc basis. The thing that has worked for us is sitting together.” “The most important aspect of One Health is the fact that that it’s impossible to control diseases that come from animals only by focusing on humans. It’s like trying to concentrate on putting out fires without ascertaining where the fires are coming from.” “Endemic diseases, despite the fact that they’re ever-present, are often the most ignored.” “A lot of the risk practices are cultural, and cultural change is very slow.” “The value of One Health is much more than the investment required to put into it. It’s one of the few things I’ve seen actually work in implementation of disease control strategies, in surveillance and in general disease control. It’s worked for Kenya and I believe it can work for all other countries.”   Links for the episode: Republic of Kenya Zoonotic Disease Unit Prioritization of Zoonotic Diseases in Kenya, 2015. Plos One.