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Topic:Pathogens

Pathogens are microorganisms, such as bacteria, viruses, fungi, and parasites, that can cause disease in horses. These microorganisms interact with the equine immune system, often leading to a range of health issues that can affect individual horses or entire populations. Pathogens can be transmitted through various routes, including direct contact, vectors, or environmental exposure. Common equine pathogens include Streptococcus equi, Equine Herpesvirus, and Strongylus vulgaris. Understanding the interactions between equine hosts and pathogens is essential for disease prevention, management, and treatment. This page compiles peer-reviewed research studies and scholarly articles that explore the identification, transmission, and impact of pathogens on equine health.
Molecular characterisation of ‘strangles’ outbreaks in the UK: the use of M-protein typing of Streptococcus equi ssp. equi.
Equine veterinary journal    August 26, 2010   Volume 43, Issue 3 359-364 doi: 10.1111/j.2042-3306.2010.00177.x
Ivens PA, Matthews D, Webb K, Newton JR, Steward K, Waller AS, Robinson C, Slater JD.Strangles is the most commonly diagnosed and important infectious disease of horses worldwide. Very little is known about the temporo-spatial and molecular epidemiology of strangles. The disease is not notifiable in the UK and there are few published data on the geographical locations of outbreaks. Objective: To investigate whether typing of a surface protein (SeM) of Streptococcus equi ssp. equi (S. equi), the causative agent of strangles, is a useful epidemiological tool. Methods: The variable region of the SeM gene was amplified from 145 isolates of S. equi by PCR and sequenced. Different S...
Characterization of vancomycin-resistant Enterococcus faecium isolated from swine in three Michigan counties.
Journal of clinical microbiology    August 25, 2010   Volume 48, Issue 11 4156-4160 doi: 10.1128/JCM.02346-09
Donabedian SM, Perri MB, Abdujamilova N, Gordoncillo MJ, Naqvi A, Reyes KC, Zervos MJ, Bartlett P.Vancomycin-resistant enterococci are a major cause of nosocomial infections but are rarely found in humans in the community and have not been identified in food animals in the United States. We evaluated a total of 360 fecal specimens from humans and their animals being raised for exhibit at three county fairs in Michigan. Fecal samples from 158 humans, 55 swine, 50 cattle, 25 horses, 57 sheep, 14 goats, and 1 llama were obtained and plated onto Enterococcosel agar containing 16 μg/ml of vancomycin. Vancomycin-resistant Enterococcus faecium (VREF) was isolated from six pigs but not from human...
Use of blood culture medium enrichment for synovial fluid culture in horses: a comparison of different culture methods.
Equine veterinary journal    August 19, 2010   Volume 42, Issue 6 541-546 doi: 10.1111/j.2042-3306.2010.00091.x
Dumoulin M, Pille F, van den Abeele AM, Boyen F, Boussauw B, Oosterlinck M, Pasmans F, Gasthuys F, Martens A.Standard methods for culturing equine synovial fluid (SF) are often unrewarding. Evidence-based information on the relative efficiency of different systems used for optimisation of isolation of microorganisms from equine SF is lacking. Objective: To compare the results of different culture systems performed in parallel on SF samples from horses clinically diagnosed with synovial sepsis. Methods: Synovial fluid specimens were collected between February 2007 and October 2008 from all horses admitted to a referral hospital that were clinically diagnosed with synovial sepsis and from control horse...
Identification of three novel superantigen-encoding genes in Streptococcus equi subsp. zooepidemicus, szeF, szeN, and szeP.
Infection and immunity    August 16, 2010   Volume 78, Issue 11 4817-4827 doi: 10.1128/IAI.00751-10
Paillot R, Darby AC, Robinson C, Wright NL, Steward KF, Anderson E, Webb K, Holden MT, Efstratiou A, Broughton K, Jolley KA, Priestnall SL....The acquisition of superantigen-encoding genes by Streptococcus pyogenes has been associated with increased morbidity and mortality in humans, and the gain of four superantigens by Streptococcus equi is linked to the evolution of this host-restricted pathogen from an ancestral strain of the opportunistic pathogen Streptococcus equi subsp. zooepidemicus. A recent study determined that the culture supernatants of several S. equi subsp. zooepidemicus strains possessed mitogenic activity but lacked known superantigen-encoding genes. Here, we report the identification and activities of three novel ...
West Nile virus range expansion into British Columbia.
Emerging infectious diseases    August 4, 2010   Volume 16, Issue 8 1251-1258 doi: 10.3201/eid1608.100483
Roth D, Henry B, Mak S, Fraser M, Taylor M, Li M, Cooper K, Furnell A, Wong Q, Morshed M.In 2009, an expansion of West Nile virus (WNV) into the Canadian province of British Columbia was detected. Two locally acquired cases of infection in humans and 3 cases of infection in horses were detected by ELISA and plaque-reduction neutralization tests. Ten positive mosquito pools were detected by reverse transcription PCR. Most WNV activity in British Columbia in 2009 occurred in the hot and dry southern Okanagan Valley. Virus establishment and amplification in this region was likely facilitated by above average nightly temperatures and a rapid accumulation of degree-days in late summer....
Pythium insidiosum: an overview.
Veterinary microbiology    July 24, 2010   Volume 146, Issue 1-2 1-16 doi: 10.1016/j.vetmic.2010.07.019
Gaastra W, Lipman LJ, De Cock AW, Exel TK, Pegge RB, Scheurwater J, Vilela R, Mendoza L.Pythium insidiosum is an oomycete pathogenic in mammals. The infection occurs mainly in tropical and subtropical areas, particularly in horses, dogs and humans. Infection is acquired through small wounds via contact with water that contains motile zoospores or other propagules (zoospores or hyphae). The disease, though described as emerging has in fact already been described since 1884. Depending on the site of entry, infection can lead to different forms of pythiosis i.e. a cutaneous, vascular, ocular, gastrointestinal and a systemic form, which is rarely seen. The infection is not contagious...
A potential role for indoleamine 2,3-dioxygenase (IDO) in Rhodococcus equi infection.
Veterinary immunology and immunopathology    July 22, 2010   Volume 138, Issue 3 174-182 doi: 10.1016/j.vetimm.2010.07.013
Heller MC, Drew CP, Jackson KA, Griffey S, Watson JL.Rhodococcus equi is a facultative intracellular bacterial pathogen of foals and immunocompromised humans that infects and proliferates within host macrophages and dendritic cells (DC). Indoleamine 2,3-dioxygenase (IDO), the initial enzyme in the tryptophan catabolism pathway, is upregulated in R. equi infected equine monocyte-derived DC and alveolar macrophages. Tryptophan requirement of R. equi for extracellular and intracellular growth was assessed. Growth of R. equi in minimal media did not require tryptophan and pharmacologic inhibition of IDO had no effect on intracellular proliferation o...
Detection of strain variation in isolates of Rhodococcus equi from an affected foal using repetitive sequence-based polymerase chain reaction. Bolton T, Kuskie K, Halbert N, Chaffin K, Healy M, Lawhon S, Jackson A, Cohen N.Rhodococcus equi is an important pathogen of foals aged 1-6 months. Evidence exists that foals are exposed to a wide diversity of R. equi strains in their environment. However, limited data are available regarding the extent to which genotypic variation exists among isolates infecting individual foals. Therefore, electrophoresis of repetitive sequence-based polymerase chain reaction (rep-PCR) amplicons in an automated microfluidics chip format was used to genotype 9 virulent R. equi isolates obtained from distinct anatomic locations in a single foal. Four of the isolates were obtained from dif...
Enrofloxacin-based therapeutic strategy for the prevention of endometritis in susceptible mares.
Journal of veterinary pharmacology and therapeutics    June 19, 2010   Volume 33, Issue 3 287-294 doi: 10.1111/j.1365-2885.2009.01135.x
González C, Moreno L, Fumuso E, García J, Rivulgo M, Confalonieri A, Sparo M, Sánchez Bruni S.Enrofloxacin (EFX) is often used empirically to prevent uterine infections in mares in order to improve efficiency on Commercial Embryo Transfer Farms. This study investigated the uterine distribution of EFX and its metabolite ciprofloxacin (CFX) in mares and assessed the minimal inhibitory concentrations (MIC) of EFX against various common pathogens as a basis for establishing a rational dosing schedule. Plasma and uterine pharmacokinetic (PK) studies were performed in two groups (n = 5) of healthy mares following intravenous (i.v.) administration of EFX at either 2.5 and at 5 mg/kg bodyweigh...
Carriage of Clostridium difficile and other enteric pathogens among a 4-H avocational cohort.
Zoonoses and public health    June 10, 2010   Volume 58, Issue 3 192-199 doi: 10.1111/j.1863-2378.2010.01338.x
McNamara SE, Abdujamilova N, Somsel P, Gordoncillo MJ, DeDecker JM, Bartlett PC.Clostridium difficile (CD), Salmonella, Campylobacter and enterohemorrhagic Escherichia coli (EHEC) are major causes of morbidity in a variety of enteric diseases in humans and animals, but subclinical carriage in both is probably more common than are clinical cases. Little is known regarding the prevalence of these pathogens in animals raised for exhibit at Michigan county fairs or the frequency with which Michigan citizens raising these animals may have been subclinically colonized. To address these issues, 361 fecal specimens from 158 humans and 203 of their farm animals were cultured for C...
Proinflammatory cytokine responses of cultured equine keratinocytes to bacterial pathogen-associated molecular pattern motifs.
Equine veterinary journal    June 9, 2010   Volume 42, Issue 4 294-303 doi: 10.2746/042516409X478523
Leise BS, Yin C, Pettigrew A, Belknap JK.Further knowledge of equine keratinocyte physiology and keratinocyte response to various stimuli is important in developing a better understanding of disease states involving the epidermis. Objective: To assess the inflammatory cytokine response of cultured equine keratinocytes to various pathogen-associated molecular pattern molecules (PAMPs) from both Gram-negative and positive bacteria likely to be present in equine sepsis. Methods: Keratinocytes were isolated from skin of 2 horses and primary cultures performed. Keratinocytes were harvested for RNA extraction after exposure to lipopolysacc...
Vector competence of Culex (Melanoconion) taeniopus for equine-virulent subtype IE strains of Venezuelan equine encephalitis virus.
The American journal of tropical medicine and hygiene    June 4, 2010   Volume 82, Issue 6 1047-1052 doi: 10.4269/ajtmh.2010.09-0556
Deardorff ER, Weaver SC.The mosquito Culex (Melanoconion) taeniopus is a proven vector of enzootic Venezuelan equine encephalitis virus (VEEV) subtype IE in Central America. It has been shown to be highly susceptible to infection by this subtype, and conversely to be highly refractory to infection by other VEEV subtypes. During the 1990s in southern coastal Mexico, two VEE epizootics in horses were attributed to subtype IE VEEV. These outbreaks were associated with VEEV strains with an altered infection phenotype for the epizootic mosquito vector, Aedes (Ochlerotatus) taeniorhynchus. To determine the infectivity for ...
In vitro antimicrobial activity of gallium maltolate against virulent Rhodococcus equi.
Veterinary microbiology    May 27, 2010   Volume 146, Issue 1-2 175-178 doi: 10.1016/j.vetmic.2010.05.027
Coleman M, Kuskie K, Liu M, Chaffin K, Libal M, Giguère S, Bernstein L, Cohen N.The objective of this study was to determine the in vitro antimicrobial activity of gallium maltolate (GaM) against Rhodococcus equi. A total of 98 virulent bacterial isolates from equine clinical cases were examined, of which 19 isolates were known to be resistant to macrolides and rifampin. Isolates were cultured with various concentrations of GaM and minimal inhibitory concentration (MIC) values were determined after 24 and 48 h. Both the MIC(50) and the MIC(90) after 24h of growth were 558 ng/mL (8 μM) and after 48 h of growth were 2230 ng/mL (32 μM). There were no apparent differences b...
Ceftiofur derivates in serum and endometrial tissue after intramuscular administration in healthy mares.
Theriogenology    May 21, 2010   Volume 74, Issue 3 466-472 doi: 10.1016/j.theriogenology.2010.02.030
Witte TS, Bergwerff AA, Scherpenisse P, Drillich M, Heuwieser W.Endometritis is one of the major problems in the horse breeding industry. The use of antibiotics for treatment of endometritis in the mare is recommended as best practice. The intrauterine application of antibiotics, however, has been under discussion over the last years because of concerns about its efficacy. The systemic use of antibiotics has been considered more effective because of its better distribution within the uterus. The objective of the present study was to determine the concentration of ceftiofur derivates in serum and endometrial tissue after intramuscular administration. Specif...
Pathological changes caused by Anoplocephala perfoliata in the equine ileocecal junction.
Veterinary research communications    May 13, 2010   Volume 34 Suppl 1 S53-S56 doi: 10.1007/s11259-010-9399-9
Pavone S, Veronesi F, Piergili Fioretti D, Mandara MT.Gastrointestinal motility disorders represent a significant cause of morbidity and mortality in horses. Previously regarded as a non-pathogenic tapeworm, Anoplocephala perfoliata has been recently associated with equine colic. In this study, pathological changes related to A. perfoliata at the ileocecal junction were investigated in 31 slaughtered horses. Our results showed a significant relationship between parasitic burden and grading of histopathological lesions in both the mucosa and submucosa. Moreover, in infested horses, hypertrophy of the circular muscle layer was determined. Finally, ...
Intra- and interhost evolutionary dynamics of equine influenza virus.
Journal of virology    May 5, 2010   Volume 84, Issue 14 6943-6954 doi: 10.1128/JVI.00112-10
Murcia PR, Baillie GJ, Daly J, Elton D, Jervis C, Mumford JA, Newton R, Parrish CR, Hoelzer K, Dougan G, Parkhill J, Lennard N, Ormond D, Moule S....Determining the evolutionary basis of cross-species transmission and immune evasion is key to understanding the mechanisms that control the emergence of either new viruses or novel antigenic variants with pandemic potential. The hemagglutinin glycoprotein of influenza A viruses is a critical host range determinant and a major target of neutralizing antibodies. Equine influenza virus (EIV) is a significant pathogen of the horse that causes periodical outbreaks of disease even in populations with high vaccination coverage. EIV has also jumped the species barrier and emerged as a novel respirator...
Evaluation of rapid antigen detection kits for diagnosis of equine rotavirus infection.
The Journal of veterinary medical science    April 28, 2010   Volume 72, Issue 9 1247-1250 doi: 10.1292/jvms.10-0064
Nemoto M, Hata H, Higuchi T, Imagawa H, Yamanaka T, Niwa H, Bannai H, Tsujimura K, Kondo T, Matsumura T.We evaluated antigen detection kits for human rotavirus with regard to their usefulness for diagnosing equine rotavirus infection. Limiting dilution analyses showed that of the seven kits investigated the Dipstick `Eiken' Rota (Dipstick) had the highest sensitivity to two serotypes of equine rotavirus. The Dipstick did not cross-react with several equine intestinal pathogens. An investigation using 249 fecal samples indicated that the sensitivity of the Dipstick was 81.9% and 47.3%, and its specificity was 98.2% and 99.0%, and its concordance rate was 92.8% and 68.3%, compared with values obta...
Haemotrophic Mycoplasma infection in horses.
Veterinary microbiology    April 24, 2010   Volume 145, Issue 3-4 351-353 doi: 10.1016/j.vetmic.2010.04.009
Dieckmann SM, Winkler M, Groebel K, Dieckmann MP, Hofmann-Lehmann R, Hoelzle K, Wittenbrink MM, Hoelzle LE.Haemotrophic mycoplasmas (HM) are parasites on the surface of red blood cells and known to infect a wide range of animals. However, there are no previous evidences of HM infections in horses. In this study HM were detected for the first time in the blood of two horses suffering from poor performance, apathy, weight loss, and anaemia. Using a HM specific PCR assay and subsequent sequencing the infective agents isolated from the blood of said horses were confirmed as closely related to the HM species Mycoplasma haemofelis and 'Candidatus Mycoplasma haemobos'.
Tuberculosis due to Mycobacterium bovis in a Camargue horse.
The Veterinary record    April 20, 2010   Volume 166, Issue 16 499-500 doi: 10.1136/vr.b4785
Keck N, Dutruel H, Smyej F, Nodet M, Boschiroli ML.No abstract available
Study of laryngopharyngeal pathology in Thoroughbred horses in southern California.
Equine veterinary journal    April 14, 2010   Volume 41, Issue 9 903-907 doi: 10.2746/042516409x448968
Diab S, Pascoe J, Shahriar M, Read D, Kinde H, Moore J, Odani J, Uzal F.There is increasing anecdotal evidence among horse owners, trainers and equine clinicians of a high prevalence of subepiglottic ulcers, suggested to have a negative effect on racing performance. Objective: To provide a prevalence study and pathological characterisation of laryngopharyngeal lesions with emphasis in the subepiglottic area and, in particular, subepiglottic ulcers. Methods: The study was carried out on 91 Thoroughbred racehorses received for post mortem examination from 4 major Southern California racetracks. The most common reason for submission was catastrophic musculoskeletal i...
IgG Endopeptidase SeMac does not Inhibit Opsonophagocytosis of Streptococcus equi Subspecies equi by Horse Polymorphonuclear Leukocytes.
The open microbiology journal    April 8, 2010   Volume 4 20-25 doi: 10.2174/1874285801004010020
Liu M, Lei B.The secreted Mac protein made by group A Streptococcus (GAS) inhibits opsonophagocytosis of GAS by human polymorphonuclear leukocytes (PMNs). This protein also has the endopeptidase activity against human immunoglobulin G (IgG), and the Cys94, His262 and Asp284 are critical for the enzymatic activity. The horse pathogen Streptococcus equi subspecies equi produces a homologue of Mac (SeMac). SeMac was characterized to determine whether SeMac has IgG endopeptidase activity and inhibits opsonophagocytosis of S. equi by horse PMNs. The gene was cloned and recombinant SeMac was overexpressed in Esc...
Cell entry of the aphthovirus equine rhinitis A virus is dependent on endosome acidification.
Journal of virology    April 7, 2010   Volume 84, Issue 12 6235-6240 doi: 10.1128/JVI.02375-09
Groppelli E, Tuthill TJ, Rowlands DJ.Equine rhinitis A virus (ERAV) is genetically closely related to foot-and-mouth disease virus (FMDV), and both are now classified within the genus Aphthovirus of the family Picornaviridae. For disease security reasons, FMDV can be handled only in high-containment facilities, but these constraints do not apply to ERAV, making it an attractive alternative for the study of aphthovirus biology. Here, we show, using immunofluorescence, pharmacological agents, and dominant negative inhibitors, that ERAV entry occurs (as for FMDV) via clathrin-mediated endocytosis and acidification of early endosomes...
In vitro predatory activity of nematophagous fungi and after passing through gastrointestinal tract of equine on infective larvae of Strongyloides westeri.
Parasitology research    April 6, 2010   Volume 107, Issue 1 103-108 doi: 10.1007/s00436-010-1841-y
Araujo JM, Araújo JV, Braga FR, Carvalho RO.Three isolates of predator fungi Duddingtonia flagrans (AC001), Monacrosporium thaumasium (NF34), and Arthrobotrys robusta (I-31) were assessed in in vitro test regarding the capacity of prey infective larvae (L(3)) Strongyloides westeri. Compared to control, without fungus, there was a significant decrease (P 0.01) in vitro test. Linear regression coefficients of treated and control groups were -0.21 for control, -0.32 for D. flagrans, -0.34 for M. thaumasium, and -0.22 for A. robusta. In the following, isolates AC001 and NF34 were assessed in vivo regarding the capacity of supporting the pa...
Pulmonary intravascular macrophages and endotoxin-induced pulmonary pathophysiology in horses. Aharonson-Raz K, Singh B.Endotoxemia causes significant mortality and morbidity in horses. The mechanisms underlying this complex pathophysiology remain unclear. Therefore, effective tools to treat endotoxemia in horses are lacking. Furthermore, the multifactorial and multiorgan pathophysiology of equine endotoxemia has not been fully addressed, especially the lung injury associated with endotoxemia. Within the context of the broader picture of endotoxemia and lung injury, we offer a perspective on the roles of pulmonary intravascular macrophages in endotoxin-induced lung inflammation in horses. L’endotoxémie est u...
A novel second instar Gasterophilus excretory/secretory antigen-based ELISA for the diagnosis of gasterophilosis in grazing horses.
Veterinary parasitology    April 1, 2010   Volume 171, Issue 3-4 314-320 doi: 10.1016/j.vetpar.2010.03.034
Sánchez-Andrade R, Cortiñas FJ, Francisco I, Sánchez JA, Mula P, Cazapal C, Vázquez L, Suárez JL, Francisco R, Arias MS, Díez-Baños P, Scala A....We have developed a novel enzyme-linked immunosorbent assay (ELISA) based on excretory/secretory antigens of second instar Gasterophilus for the diagnosis of gasterophilosis in grazing horses. Between January 2007 and January 2009, two experiments were carried out on free-ranging horses in northwest Spain. During the first year, monthly blood samples were collected from a herd of 25 horses. In the second year, a monthly serological survey was conducted for a total of 398 different horses. All the sera were analyzed by ELISA using excretory/secretory antigens from Gasterophilus intestinalis (Gp...
Check list of the helminths of equines in Turkey.
Turkiye parazitolojii dergisi    March 27, 2010   Volume 34, Issue 1 40-44 
Gürler AT, Bölükbaş CS, Açici M, Umur S.Helminths of equines are one of the most important agents of parasitic diseases. Therefore, many studies have been conducted on helminths of equines in Turkey. In this article, a check list and prevalence rates of helminths of equines in Turkey have been given.
Examination of equine glandular stomach lesions for bacteria, including Helicobacter spp by fluorescence in situ hybridisation.
BMC microbiology    March 19, 2010   Volume 10 84 doi: 10.1186/1471-2180-10-84
Husted L, Jensen TK, Olsen SN, Mølbak L.The equine glandular stomach is commonly affected by erosion and ulceration. The aim of this study was to assess whether bacteria, including Helicobacter, could be involved in the aetiology of gastric glandular lesions seen in horses. Results: Stomach lesions, as well as normal appearing mucosa were obtained from horses slaughtered for human consumption. All samples were tested for urease activity using the Pyloritek assay, while mucosal bacterial content was evaluated using Fluorescence In Situ Hybridisation. In selected sub samples, bacteria characterisation was pursued further by cloning an...
Identification of Rhodococcus equi lipids recognized by host cytotoxic T lymphocytes.
Microbiology (Reading, England)    March 18, 2010   Volume 156, Issue Pt 6 1836-1847 doi: 10.1099/mic.0.035915-0
Harris SP, Fujiwara N, Mealey RH, Alperin DC, Naka T, Goda R, Hines SA.Immune adult horses have CD8(+) cytotoxic T lymphocytes (CTLs) that recognize and lyse Rhodococcus equi-infected cells in an equine lymphocyte alloantigen (ELA)-A [classical major histocompatibility complex (MHC) class I]-unrestricted fashion. As protein antigens are MHC class I-restricted, the lack of restriction suggests that the bacterial antigens being recognized by the host are not proteins. The goals of this study were to test the hypothesis that these CTLs recognize unique R. equi cell-wall lipids related to mycobacterial lipids. Initial experiments showed that treatment of soluble R. e...
Characterization of the haem-uptake system of the equine pathogen Streptococcus equi subsp. equi.
Microbiology (Reading, England)    March 11, 2010   Volume 156, Issue Pt 6 1824-1835 doi: 10.1099/mic.0.036087-0
Meehan M, Burke FM, Macken S, Owen P.Streptococcus equi possesses a haem-uptake system homologous to that of Streptococcus pyogenes and Streptococcus zooepidemicus. The system consists of two ligand-binding proteins (Shr and Shp) and proteins (HtsA-C) with homology to an ABC transporter. The haem-uptake system of S. equi differs from that of S. pyogenes and S. zooepidemicus in that Shr is truncated by two-thirds. This study focused on the SeShr, SeShp and SeHtsA proteins of S. equi. Analysis of shr, shp and shphtsA knockout mutants showed that all three proteins were expressed in vitro and that expression was upregulated under co...
Detection of Streptococcus dysgalactiae subsp. equisimilis in equine nasopharyngeal swabs by PCR.
Journal of veterinary science    March 3, 2010   Volume 11, Issue 1 67-72 doi: 10.4142/jvs.2010.11.1.67
Preziuso S, Laus F, Tejeda AR, Valente C, Cuteri V.Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this stu...
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