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Topic:Microbiology

Microbiology and horses explores the interactions between microorganisms and equine hosts, focusing on the role of bacteria, viruses, fungi, and other microbes in horse health and disease. This field examines the microbial flora present in various equine environments, including the gastrointestinal tract, skin, and respiratory system, and investigates how these microorganisms influence equine physiology and pathology. Research in this area includes studies on microbial infections that affect horses, the development of antimicrobial resistance, and the impact of probiotics and prebiotics on equine health. This page compiles peer-reviewed research studies and scholarly articles that investigate the diversity, dynamics, and implications of microbial communities in horses, as well as the strategies for managing microbial-related diseases.
Comparison of Fecal Microbiota of Mongolian and Thoroughbred Horses by High-throughput Sequencing of the V4 Region of the 16S rRNA Gene.
Asian-Australasian journal of animal sciences    December 1, 2015   Volume 29, Issue 9 1345-1352 doi: 10.5713/ajas.15.0587
Zhao Y, Li B, Bai D, Huang J, Shiraigo W, Yang L, Zhao Q, Ren X, Wu J, Bao W, Dugarjaviin M.The hindgut of horses is an anaerobic fermentative chamber for a complex and dynamic microbial population, which plays a critical role in health and energy requirements. Research on the gut microbiota of Mongolian horses has not been reported until now as far as we know. Mongolian horse is a major local breed in China. We performed high-throughput sequencing of the 16S rRNA genes V4 hypervariable regions from gut fecal material to characterize the gut microbiota of Mongolian horses and compare them to the microbiota in Thoroughbred horses. Fourteen Mongolian and 19 Thoroughbred horses were use...
Three new serotypes of Rhodococcus equi in Prescott’s serotyping system – Short communication.
Acta veterinaria Hungarica    November 10, 2015   Volume 63, Issue 3 265-270 doi: 10.1556/004.2015.024
Makrai L, Fodor L, Hajtós I, Varga J, Dénes B.Three new serotypes were found among Rhodococcus equi strains, which could not be assigned into any of the seven serotypes of Prescott's system. Fortythree R. equi strains out of 44 previously nontypable ones isolated in Hungary could be allocated into one of the three new serotypes using the agar gel immunodiffusion (AGID) test. The three new suggested serotypes are serotype 8 (proposed reference strain: HNCMB-138003), serotype 9 (proposed reference strain: HNCMB-138004) and serotype 10 (proposed reference strain: HNCMB-138005). Hyperimmune sera produced in rabbits against the new serotypes a...
Occurrence and distribution of Malassezia species on skin and external ear canal of horses.
Mycoses    November 9, 2015   Volume 59, Issue 1 28-33 doi: 10.1111/myc.12430
Shokri H.The aim of this study was to investigate the prevalence of Malassezia species from the body skin and external ear canal of healthy horses. The samples were obtained by scraping the skin surface from the nose, groin and dorsum and swabbing from the external ear canal of 163 animals, and then incubated on sabouraud dextrose agar and modified Dixon agar. Malassezia species were isolated from 34.9% of horses. The percentages of Malassezia species were 64.3% for Arab, 35.7% for Persian, 35.4% for Thoroughbred and 27.1% for Turkmen breeds. The greatest abundance of Malassezia species was found in th...
Fecal Ciliate Composition of Domestic Horses (Equus caballus Linnaeus, 1758) Living in Kyrgyzstan.
Zootaxa    November 3, 2015   Volume 4039, Issue 1 145-156 doi: 10.11646/zootaxa.4039.1.6
Gürelli G, Canbulat S, Aldayarov N.Species composition and distribution of intestinal ciliates were investigated in the feces from 15 domestic horses living in Bishkek, Kyrgyzstan. Twenty-three species belonging to 14 genera were identified. This is the first study on intestinal ciliates in domestic horses living in Kyrgyzstan. The mean number of ciliates was 14.1 ± 6.8 x10(4) cells ml(-1) of feces and the mean number of ciliate species per host was 6.0 ± 3.2. No endemic or new species were detected. Blepharocorys was the major genus as these ciliates were detected in high proportions. In contrast Holophryoides, Allantosoma w...
Trends in antifungal susceptibility and virulence of Candida spp. from the nasolacrimal duct of horses.
Medical mycology    October 18, 2015   Volume 54, Issue 2 147-154 doi: 10.1093/mmy/myv090
Brilhante RS, Bittencourt PV, Castelo-Branco Dde S, de Oliveira JS, Alencar LP, Cordeiro Rde A, Pinheiro M, Nogueira-Filho EF, Pereira-Neto Wde A....This was a cross-sectional study to investigate the antifungal susceptibility and production of virulence factors in strains of Candida isolated from the outlet and the lumen of the nasolacrimal duct of horses in the state of Ceará, Brazil. The samples were obtained from 103 horses. Sterile cotton swabs were used to collect the material from the outlet of the nasolacrimal duct and urethral probes, for the instillation of 2 ml of saline solution, were used to collect samples from the lumen of the nasolacrimal duct. A total of 77 Candida isolates were obtained, with C. famata, C. tropicalis, C....
Methicillin-resistant Staphylococcus aureus ulcerative keratitis in a Thoroughbred racehorse.
Journal of equine science    September 30, 2015   Volume 26, Issue 3 95-98 doi: 10.1294/jes.26.95
Kuroda T, Kinoshita Y, Niwa H, Mizobe F, Ueno T, Kuwano A, Hatazoe T, Hobo S.We report the first case of methicillin-resistant Staphylococcus aureus (MRSA) keratitis in a racehorse. A 5-year-old mare developed punctate keratitis after racing. The corneal ulcer continued to expand despite ophthalmic antimicrobial therapy. On day 6, a conjunctival graft surgery was performed. The mare was euthanized, following colitis and laminitis development on day 10. MRSA was isolated from the corneal swab taken at the time of euthanasia. Immunohistochemical analysis demonstrated gram-positive and anti-S. aureus monoclonal antibody-positive cocci infiltration of the corneal stroma; a...
Effect of starch source (corn, oats or wheat) and concentration on fermentation by equine faecal microbiota in vitro.
Journal of applied microbiology    September 28, 2015   Volume 119, Issue 5 1234-1244 doi: 10.1111/jam.12927
Harlow BE, Donley TM, Lawrence LM, Flythe MD.The goal was to determine the effect of starch source (corn, oats and wheat) and concentration on: (i) total amylolytic bacteria, Group D Gram-positive cocci (GPC), lactobacilli and lactate-utilizing bacteria, and (ii) fermentation by equine microbiota. Results: When faecal washed cell suspensions were incubated with any substrate amylolytics increased over time. However, at 24 h there were 10 and 1000-fold more amylolytics with corn than wheat or oats respectively. Predominant amylolytics isolated were Enterococcus faecalis (corn, wheat) and Streptococcus bovis (oats). GPC increased with any ...
Clinical Research Abstracts of the British Equine Veterinary Association Congress 2015.
Equine veterinary journal    September 17, 2015   Volume 47 Suppl 48 3 doi: 10.1111/evj.12486_6
Allen JL, Herbert G, Muscatello G, Gilkerson JR.Bronchopneumonia caused by Rhodococcus equi is an important disease of young horses throughout the world. Although early diagnosis and treatment improves the prognosis, this also increases the amount of antimicrobial usage and therefore increases the likelihood of resistance developing. Objective: The aim of this study was to determine the level of resistance to commonly prescribed antimicrobial agents of 97 virulent Rhodococcus equi isolates. Methods: Analysis of archived samples. Methods: Virulent Rhodococcus equi isolates were collected between 1991 and 2014 from clinically affected horses ...
Complete genome sequences of T5-related Escherichia coli bacteriophages DT57C and DT571/2 isolated from horse feces.
Archives of virology    September 9, 2015   Volume 160, Issue 12 3133-3137 doi: 10.1007/s00705-015-2582-0
Golomidova AK, Kulikov EE, Prokhorov NS, Guerrero-Ferreira RC, Ksenzenko VN, Tarasyan KK, Letarov AV.We report the complete genome sequencing of two Escherichia coli T5-related bacteriophages, DT57C and DT571/2, isolated from the same specimen of horse feces. These two isolates share 96% nucleotide sequence identity and can thus be considered representatives of the same novel species within the genus T5likevirus. The observed variation in the ltfA gene of these phages, resulting from a recent recombination event, may explain the observed host-range differences, suggesting that a modular mechanism makes a significant contribution to the short-term evolution (or adaptation) of T5-like phage gen...
Anti-biofilm activity of ultrashort cinnamic acid peptide derivatives against medical device-related pathogens.
Journal of peptide science : an official publication of the European Peptide Society    August 27, 2015   Volume 21, Issue 10 770-778 doi: 10.1002/psc.2805
Laverty G, McCloskey AP, Gorman SP, Gilmore BF.The threat of antimicrobial resistance has placed increasing emphasis on the development of innovative approaches to eradicate multidrug-resistant pathogens. Biofilm-forming microorganisms, for example, Staphylococcus epidermidis and Staphylococcus aureus, are responsible for increased incidence of biomaterial infection, extended hospital stays and patient morbidity and mortality. This paper highlights the potential of ultrashort tetra-peptide conjugated to hydrophobic cinnamic acid derivatives. These peptidomimetic molecules demonstrate selective and highly potent activity against resistant b...
Effects of transport, fasting and anaesthesia on the faecal microbiota of healthy adult horses.
Equine veterinary journal    August 18, 2015   Volume 48, Issue 5 595-602 doi: 10.1111/evj.12479
Schoster A, Mosing M, Jalali M, Staempfli HR, Weese JS.The intestinal microbiota is important for health and disease. Factors that disturb the equine intestinal microbiota need further investigation. Objective: To determine the effects of transport, fasting and anaesthesia on the faecal microbiota of healthy adult horses using next-generation sequencing. Methods: Experimental trial. Methods: Faecal samples were taken from 8 horses at baseline, after transport, 12 h of fasting and 24, 48 and 72 h after a 6 h anaesthesia. Next generation sequencing of the V4 region of the 16S rRNA gene was used to assess the microbial composition of faeces. Alpha...
Virulence Factor Genes Detected in the Complete Genome Sequence of Corynebacterium uterequi DSM 45634, Isolated from the Uterus of a Maiden Mare.
Genome announcements    July 30, 2015   Volume 3, Issue 4 e00783-15 doi: 10.1128/genomeA.00783-15
Rückert C, Kriete M, Jaenicke S, Winkler A, Tauch A.The complete genome sequence of the type strain Corynebacterium uterequi DSM 45634 from an equine urogenital tract specimen comprises 2,419,437 bp and 2,163 protein-coding genes. Candidate virulence factors are homologs of DIP0733, DIP1281, and DIP1621 from Corynebacterium diphtheriae and of sialidase precursors from Trueperella pyogenes and Chlamydia trachomatis.
Genome specialization and decay of the strangles pathogen, Streptococcus equi, is driven by persistent infection.
Genome research    July 9, 2015   Volume 25, Issue 9 1360-1371 doi: 10.1101/gr.189803.115
Harris SR, Robinson C, Steward KF, Webb KS, Paillot R, Parkhill J, Holden MT, Waller AS.Strangles, the most frequently diagnosed infectious disease of horses worldwide, is caused by Streptococcus equi. Despite its prevalence, the global diversity and mechanisms underlying the evolution of S. equi as a host-restricted pathogen remain poorly understood. Here, we define the global population structure of this important pathogen and reveal a population replacement in the late 19th or early 20th Century. Our data reveal a dynamic genome that continues to mutate and decay, but also to amplify and acquire genes despite the organism having lost its natural competence and become host-rest...
Molecular studies on European equine isolates of Leptospira interrogans serovars Bratislava and Muenchen. Arent Z, Gilmore C, Brem S, Ellis WA.Strains of Leptospira interrogans belonging to two very closely related serovars – Bratislava and Muenchen – are known to cause widespread infection of the horse population in many parts of the world. Conventional serological typing of isolates has been unable to differentiate between wildlife, pig, dog and possibly horse maintained isolates and therefore has been unable to provide further insight into their diversity and the relationship between them. Twenty-one such European isolates of serovar Bratislava and Muenchen were examined by restriction endonuclease analysis and multiple-locus ...
Equine recurrent uveitis: Human and equine perspectives.
Veterinary journal (London, England : 1997)    July 3, 2015   Volume 206, Issue 1 22-29 doi: 10.1016/j.tvjl.2015.06.017
Malalana F, Stylianides A, McGowan C.Equine recurrent uveitis (ERU) is a spontaneous disease characterised by repeated episodes of intraocular inflammation. The epidemiology of ERU has not been fully elucidated, but the condition appears to be much more common in horses than is recurrent uveitis in humans, especially in certain breeds and geographical regions. Both humans and horses show a similarly altered immune response and a marked autoimmune response as the primary disease pathophysiology. However, an inciting cause is not always clear. Potential inciting factors in horses include microbial agents such as Leptospira spp. Mic...
Prevalence of Streptococcus dysgalactiae subsp. equisimilis and S. equi subsp. zooepidemicus in a sample of healthy dogs, cats and horses.
New Zealand veterinary journal    June 18, 2015   Volume 63, Issue 5 265-271 doi: 10.1080/00480169.2015.1016133
Acke E, Midwinter AC, Lawrence K, Gordon SJ, Moore S, Rasiah I, Steward K, French N, Waller A.To estimate the prevalence of β-haemolytic Lancefield group C streptococci in healthy dogs, cats and horses; to determine if frequent contact with horses was associated with isolation of these species from dogs and cats; and to characterise recovered S. equi subsp. zooepidemicus isolates by multilocus sequence typing. Methods: Oropharyngeal swabs were collected from 197 dogs and 72 cats, and nasopharyngeal swabs from 93 horses. Sampling was carried out at the Massey University Veterinary Teaching Hospital, on sheep and beef farms or on premises where horses were present. All animals were heal...
Cladophialophora bantiana as an Emerging Pathogen in Animals: Case Report of Equine Endometritis and Review of the Literature.
Journal of clinical microbiology    June 17, 2015   Volume 53, Issue 9 3047-3053 doi: 10.1128/JCM.00728-15
Rantala M, Attia S, Koukila-Kähkölä P, de Hoog S, Anttila M, Katila T.We present an unusual equine endometritis case associated with Cladophialophora bantiana in a 15-year-old mare. The mare displayed infertility and uterine fluid accumulation with numerous black, hairy granules. Microscopically, the fluid revealed numerous septate, dark fungal hyphae and conidia in chains. Culture yielded C. bantiana (CBS 138271); the species was confirmed by internal transcribed spacer (ITS) sequencing. Treatment was unsuccessful. C. bantiana causes cerebral phaeohyphomycosis in humans, while animal cases are rare. Animal cases are reviewed.
Effects of age on follicular fluid exosomal microRNAs and granulosa cell transforming growth factor-β signalling during follicle development in the mare.
Reproduction, fertility, and development    May 7, 2015   Volume 27, Issue 6 897-905 doi: 10.1071/RD14452
da Silveira JC, Winger QA, Bouma GJ, Carnevale EM.Age-related decline in fertility is a consequence of low oocyte number and/or low oocyte competence resulting in pregnancy failure. Transforming growth factor (TGF)-β signalling is a well-studied pathway involved in follicular development and ovulation. Recently, small non-coding RNAs, namely microRNAs (miRNAs), have been demonstrated to regulate several members of this pathway; miRNAs are secreted inside small cell-secreted vesicles called exosomes. The overall goal of the present study was to determine whether altered exosome miRNA content in follicular fluid from old mares is associated wi...
Limited similarity between plasmids encoding CTX-M-1 β-lactamase in Escherichia coli from humans, pigs, cattle, organic poultry layers and horses in Denmark.
Journal of global antimicrobial resistance    May 7, 2015   Volume 3, Issue 2 132-136 doi: 10.1016/j.jgar.2015.03.009
Jakobsen L, Bortolaia V, Bielak E, Moodley A, Olsen SS, Hansen DS, Frimodt-Møller N, Guardabassi L, Hasman H.CTX-M-1 is a common extended-spectrum β-lactamase (ESBL) in Escherichia coli from animals and is often detected among human clinical isolates. The objective of this study was to investigate the epidemiological relationship between CTX-M-1-producing E. coli isolated from patients and animals in Denmark between 2006 and 2010. In total, 65 CTX-M-1-producing isolates from patients (n=22), pigs (n=21), cattle (n=4), organic poultry layers (n=3) and horses (n=15) were typed by pulsed-field gel electrophoresis (PFGE). Plasmids harbouring bla were characterised by S1 PFGE, PCR-based replicon typing, ...
Charting stormy waters: A commentary on the nomenclature of the equine pathogen variously named Prescottella equi, Rhodococcus equi and Rhodococcus hoagii.
Equine veterinary journal    April 24, 2015   Volume 47, Issue 5 508-509 doi: 10.1111/evj.12399
Goodfellow M, Sangal V, Jones AL, Sutcliffe IC.No abstract available
Efficacy of Mechanical versus Non-Mechanical Sterile Preoperative Skin Preparation With Chlorhexidine Gluconate 4% Solution.
Veterinary surgery : VS    April 24, 2015   Volume 44, Issue 5 648-652 doi: 10.1111/vsu.12335
Davids BI, Davidson MJ, TenBroeck SH, Colahan PT, Oli MW.To compare the efficacy of sterile preoperative skin antisepsis using either a 5-minute mechanical preparation or 5-minute non-mechanical preparation with chlorhexidine gluconate 4% solution. Methods: Matched design, ANOVA. Methods: Healthy adult Thoroughbred horses (n = 30). Methods: Each horse had both surgical preparation methods randomly assigned to identical sites on the left or right upper thigh. Prepared sites were sampled and cultured for bacteria after each preparation step. Results: Mechanical and non-mechanical preparation techniques significantly reduced bacteria isolated from ...
Equid herpesvirus type 1 activates platelets.
PloS one    April 23, 2015   Volume 10, Issue 4 e0122640 doi: 10.1371/journal.pone.0122640
Stokol T, Yeo WM, Burnett D, DeAngelis N, Huang T, Osterrieder N, Catalfamo J.Equid herpesvirus type 1 (EHV-1) causes outbreaks of abortion and neurological disease in horses. One of the main causes of these clinical syndromes is thrombosis in placental and spinal cord vessels, however the mechanism for thrombus formation is unknown. Platelets form part of the thrombus and amplify and propagate thrombin generation. Here, we tested the hypothesis that EHV-1 activates platelets. We found that two EHV-1 strains, RacL11 and Ab4 at 0.5 or higher plaque forming unit/cell, activate platelets within 10 minutes, causing α-granule secretion (surface P-selectin expression) and pl...
Rapid and accurate identification of Streptococcus equi subspecies by MALDI-TOF MS.
Systematic and applied microbiology    April 23, 2015   Volume 38, Issue 5 315-322 doi: 10.1016/j.syapm.2015.02.010
Kudirkiene E, Welker M, Knudsen NR, Bojesen AM.Streptococcus equi includes very important animal and human pathogens. S. equi subsp. equi (SEE) is a highly pathogenic equine specific subspecies, while S. equi subsp. zooepidemicus (SEZ) and S. equi subsp. ruminatorum are opportunistic pathogens of various animal species and humans. Due to great phenotypic and sequence similarity between three subspecies their discrimination remains difficult. In this study, we aimed to design and validate a novel, Superspectra based, MALDI-TOF MS approach for reliable, rapid and cost-effective identification of SEE and SEZ, the most frequent S. equi subspec...
Use of loop-mediated isothermal amplification to detect six groups of pathogens causing secondary lower respiratory bacterial infections in horses.
Microbiology and immunology    April 8, 2015   Volume 59, Issue 6 365-370 doi: 10.1111/1348-0421.12257
Kinoshita Y, Niwa H, Katayama Y.Microbial substitution occasionally occurs following the administration of antimicrobials to horses that have pneumonia or pleuropneumonia. Four specific loop-mediated isothermal amplification (LAMP) assays were developed to detect some equine respiratory pathogens, namely strains of the Bacteroides-Prevotella group, Klebsiella pneumoniae, Stenotrophomonas maltophilia, and Staphylococcus aureus. These four LAMP assays and two previously published LAMP assays targeting Escherichia coli or Pseudomonas aeruginosa were used on clinical respiratory specimens and a high accordance found between the ...
Lactobacillus ruminis strains cluster according to their mammalian gut source.
BMC microbiology    April 1, 2015   Volume 15 80 doi: 10.1186/s12866-015-0403-y
O' Donnell MM, Harris HM, Lynch DB, Ross RP, O'Toole PW.Lactobacillus ruminis is a motile Lactobacillus that is autochthonous to the human gut, and which may also be isolated from other mammals. Detailed characterization of L. ruminis has previously been restricted to strains of human and bovine origin. We therefore sought to expand our bio-bank of strains to identify and characterise isolates of porcine and equine origin by comparative genomics. Results: We isolated five strains from the faeces of horses and two strains from pigs, and compared their motility, biochemistry and genetic relatedness to six human isolates and three bovine isolates incl...
Characterization and comparison of the bacterial microbiota in different gastrointestinal tract compartments in horses.
Veterinary journal (London, England : 1997)    March 23, 2015   Volume 205, Issue 1 74-80 doi: 10.1016/j.tvjl.2015.03.018
Costa MC, Silva G, Ramos RV, Staempfli HR, Arroyo LG, Kim P, Weese JS.The advance of new sequencing technologies has allowed more comprehensive characterization of complex microbial communities, including the ones inhabiting the intestinal tract. The presence of extreme environmental filters, such as low pH, digestive enzymes and anaerobic conditions along the tract, acts on the selection of unique bacteria in each compartment. The intestinal microbiota has an enormous impact on the maintenance of health. However, data about the bacteria present in the different intestinal compartments of horses are sparse. In this study, high throughput sequencing was used to c...
PinR mediates the generation of reversible population diversity in Streptococcus zooepidemicus.
Microbiology (Reading, England)    February 20, 2015   Volume 161, Issue Pt 5 1105-1112 doi: 10.1099/mic.0.000057
Steward KF, Harrison T, Robinson C, Slater J, Maskell DJ, Harris SR, Holden MTG, Waller AS.Opportunistic pathogens must adapt to and survive in a wide range of complex ecosystems. Streptococcus zooepidemicus is an opportunistic pathogen of horses and many other animals, including humans. The assembly of different surface architecture phenotypes from one genotype is likely to be crucial to the successful exploitation of such an opportunistic lifestyle. Construction of a series of mutants revealed that a serine recombinase, PinR, inverts 114 bp of the promoter of SZO_08560, which is bordered by GTAGACTTTA and TAAAGTCTAC inverted repeats. Inversion acts as a switch, controlling the tra...
Structural characterisation of the virulence-associated protein VapG from the horse pathogen Rhodococcus equi.
Veterinary microbiology    February 9, 2015   Volume 179, Issue 1-2 42-52 doi: 10.1016/j.vetmic.2015.01.027
Okoko T, Blagova EV, Whittingham JL, Dover LG, Wilkinson AJ.Virulence and host range in Rhodococcus equi depends on the variable pathogenicity island of their virulence plasmids. Notable gene products are a family of small secreted virulence-associated proteins (Vaps) that are critical to intramacrophagic proliferation. Equine-adapted strains, which cause severe pyogranulomatous pneumonia in foals, produce a cell-associated VapA that is necessary for virulence, alongside five other secreted homologues. In the absence of biochemical insight, attention has turned to the structures of these proteins to develop a functional hypothesis. Recent studies have ...
Changes in the equine fecal microbiota associated with the use of systemic antimicrobial drugs.
BMC veterinary research    February 3, 2015   Volume 11 19 doi: 10.1186/s12917-015-0335-7
The intestinal tract is a rich and complex environment and its microbiota has been shown to have an important role in health and disease in the host. Several factors can cause disruption of the normal intestinal microbiota, including antimicrobial therapy, which is an important cause of diarrhea in horses. This study aimed to characterize changes in the fecal bacterial populations of healthy horses associated with the administration of frequently used antimicrobial drugs. Results: Twenty-four adult mares were assigned to receive procaine penicillin intramuscularly (IM), ceftiofur sodium IM, tr...
Colostrum hexasaccharide, a novel Staphylococcus aureus quorum-sensing inhibitor.
Antimicrobial agents and chemotherapy    February 2, 2015   Volume 59, Issue 4 2169-2178 doi: 10.1128/AAC.03722-14
Srivastava A, Singh BN, Deepak D, Rawat AK, Singh BR.The discovery of quorum-sensing (QS) systems regulating antibiotic resistance and virulence factors (VFs) has afforded a novel opportunity to prevent bacterial pathogenicity. Dietary molecules have been demonstrated to attenuate QS circuits of bacteria. But, to our knowledge, no study exploring the potential of colostrum hexasaccharide (CHS) in regulating QS systems has been published. In this study, we analyzed CHS for inhibiting QS signaling in Staphylococcus aureus. We isolated and characterized CHS from mare colostrum by high-performance thin-layer chromatography (HPTLC), reverse-phase hig...
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