Antimicrobial‐resistant Enterobacteriaceae recovered from companion animal and livestock environments

Antimicrobial‐resistant bacteria represent an important concern impacting both veterinary medicine and public health. The rising prevalence of extended‐spectrum beta‐lactamase (ESBL), AmpC beta‐lactamase, carbapenemase (CRE) and fluoroquinolone‐resistant Enterobacteriaceae continually decreases the efficiency of clinically important antibiotics. Moreover, the potential for zoonotic transmission of antibiotic‐resistant enteric bacteria increases the risk to public health. Our objective was to estimate the prevalence of specific antibiotic‐resistant bacteria on human contact surfaces in various animal environments. Environmental surface samples were collected from companion animal shelters, private equine facilities, dairy farms, livestock auction markets and livestock areas of county fairs using electrostatic cloths. Samples were screened for Enterobacteriaceae expressing AmpC, ESBL, CRE or fluoroquinolone resistance using selective media. Livestock auction markets and county fairs had higher levels of bacteria expressing both cephalosporin and fluoroquinolone resistance than did equine, dairy, and companion animal environments. Equine facilities harboured more bacteria expressing cephalosporin resistance than companion animal shelters, but less fluoroquinolone resistance. The regular use of extended‐spectrum cephalosporins in livestock populations could account for the increased levels of cephalosporin resistance in livestock environments compared to companion animal and equine facilities. Human surfaces, as well as shared human and animal surfaces, were contaminated with resistant bacteria regardless of species environment. Detecting these bacteria on common human contact surfaces suggests that the environment can serve as a reservoir for the zoonotic transmission of antibiotic‐resistant bacteria and resistance genes. Identifying interventions to lower the prevalence of antibiotic‐resistant bacteria in animal environments will protect both animal and public health.     

Antimicrobial resistance trends among canine Escherichia coli isolates obtained from clinical samples in the northeastern USA, 2004–2011

Our objectives were to describe the antimicrobial susceptibility of Escherichia coli isolates from dogs in the northeastern USA and to identify temporal trends in resistance to selected antimicrobial agents. Data were collected retrospectively for all canine E. coli isolates from clinical samples submitted to Cornell University’s Animal Health Diagnostic Center between January 1, 2004 and December 31, 2011. Antimicrobial susceptibility testing was performed on 3519 canine E. coli isolates; frequency of resistance to each agent ranged from 0.4% (amikacin) to 34.3% (ampicillin). No trends were evident among urinary isolates, but cephalosporin resistance remained consistently high. Among non-urinary isolates, there was evidence of a significantly increasing trend in prevalence of resistance to several agents, including cephalosporins, enrofloxacin, and tetracycline. These data suggest that some of the most commonly used antimicrobial agents in companion animal practice are becoming less effective against canine E. coli infections outside the urinary tract.     

The spread and antimicrobial resistance of Staphylococcus aureus in South African dairy herds – A review

Staphylococcus aureus is internationally recognised as a principal agent of mastitis and the foremost reason for economic loss in the dairy industry. The limited data available on organism-specific antibiotic resistance surveillance in dairy cattle have stimulated the need for such a review article. The objective of this study was to review relevant literature on antimicrobial resistance of mastitis-causing staphylococci isolated from dairy cows in South Africa compared to other countries. Factors relating to the incidence of mastitis and treatment strategies in terms of the One Health concept and food security were included. The Web of Science (all databases) and relevant websites were used, and articles not written in English were excluded. The incidence of mastitis varied between South Africa and other countries. Antimicrobial resistance patterns caused by S. aureus also varied in regions within Southern Africa and those of other countries although some similarities were shown. Antimicrobial resistance differed between S. aureus bacteria that were maltose positive and negative (an emerging pathogen). The results highlighted the importance of the availability of organism-specific surveillance data of the incidence of mastitis and antibiotic resistance for specific countries and within similar climatic conditions. Accurate knowledge about whether a specific pathogen is resistant to an antibiotic within a certain climate, country, area or farm should reduce the incidence of unnecessary or incorrect treatment with antibiotics. This should enable dairy farmers to deal with these organisms in a more effective manner. Therefore such research should be ongoing.     

Antibiotic resistant Escherichia coli in southeastern Australian pig herds and implications for surveillance

To investigate public health implications of antibiotics to control post‐weaning scours, we surveyed 22 commercial pig herds in southeastern Australia. Fifty faecal samples per herd were collected from pre‐ and post‐weaned piglets. Presumptive Escherichia coli isolates were confirmed by MALDI‐TOF MS. Isolates (n = 325) were screened for susceptibility to 19 veterinary antibiotics using MIC broth microdilution. All 325 E. coli isolates underwent further testing against 27 antibiotics used in human medicine and were screened for ETEC adhesin and enterotoxin genes (F4 (K88), F5 (K99), F6 (987P), F18, F41, STa, STb, Stx2e and LT) by multiplex PCR. Isolates identified as phenotypically resistant to third‐generation cephalosporin (3GC) and aminoglycoside antibiotics were screened by multiplex PCR/reverse line blot to detect common β‐lactam and aminoglycosides resistance genes, confirmed by sequencing. Twenty (6.1%) of the E. coli isolates were resistant to 3GC antibiotics and 24 (7.4%) to the aminoglycoside antibiotic gentamicin. Genetic analysis revealed six different extended spectrum β‐lactamase (ESBL) genes (blaCTX‐M‐1, ‐14, ‐15, ‐27, blaSHV‐12 and blaCMY‐2‐like genes), four of which have not been previously reported in Australian pigs. Critically, the prevalence of 3GC resistance was higher in non‐pathogenic (non‐ETEC) isolates and those from clinically normal (non‐diarrhoeal) samples. This highlights the importance of non‐ETECE. coli as reservoirs of antimicrobial resistance genes in piglet pens. Antimicrobial resistance surveillance in pig production focused on diagnostic specimens from clinically‐affected animals might be potentially misleading. We recommend that surveillance for emerging antimicrobial resistance such as to 3GC antibiotics should include clinically healthy pigs.     

Preliminary study of the anaerobic bacteria isolated from subgingival plaque from sheep

A preliminary study was made to determine the genera of cultivable anaerobic bacteria which could be isolated from subgingival plaque of sheep. Samples were taken from 10 sheep on farms with a known record of broken mouth periodontitis. For assessment of the sampling technique, samples were also taken from freshly exposed tooth roots in killed sheep. The bacteria isolated on several selective and non-selective media were identified to genus level by a combination of Gram reaction, colony morphology and gas chromatographic analysis of volatile and non-volatile fatty acid metabolic end products. At least 10 different genera were isolated and these findings are discussed in relation to the bacteriology of human periodontitis and recent studies of sheep broken mouth periodontitis.     

Antimicrobial susceptibility of Escherichia coli F4, Pasteurella multocida,and Streptococcus suis isolates from a diagnostic veterinary laboratory and recommendations for a surveillance system

Antimicrobial susceptibility data on Escherichia coli F4, Pasteurella multocida, and Streptococcus suis isolates from Ontario swine (January 1998 to October 2010) were acquired from a comprehensive diagnostic veterinary laboratory in Ontario, Canada. In relation to the possible development of a surveillance system for antimicrobial resistance, data were assessed for ease of management, completeness, consistency, and applicability for temporal and spatial statistical analyses. Limited farm location data precluded spatial analyses and missing demographic data limited their use as predictors within multivariable statistical models. Changes in the standard panel of antimicrobials used for susceptibility testing reduced the number of antimicrobials available for temporal analyses. Data consistency and quality could improve over time in this and similar diagnostic laboratory settings by encouraging complete reporting with sample submission and by modifying database systems to limit free-text data entry. These changes could make more statistical methods available for disease surveillance and cluster detection.     

An investigation of exudative epidermitis (greasy pig disease) and antimicrobial resistance patterns of Staphylococcus hyicus and Staphylococcus aureus isolated from clinical cases

Exudative epidermitis (EE) is a common skin disease of young pigs, caused mainly by Staphylococcus hyicus. Increased prevalence of EE and poor response to treatment are reported. Common strategies used by Ontario pork producers to treat pigs with EE were determined using a survey. Injection of penicillin G was reported as the most common parenteral antibiotic choice. Antimicrobial resistance patterns of S. hyicus and Staphylococcus aureus isolated from clinical cases (30 herds with samples from approximately 6 pigs per farm) showed that 97% of S. hyicus isolates were resistant to penicillin G and ampicillin; 71% of these isolates were resistant to ceftiofur. Similar resistance was noted among S. aureus isolates. Antimicrobial resistance has become a problem in the treatment of EE in Ontario.     

Multidrug-resistant <Emphasis Type="Italic">Salmonellae</Emphasis> isolated in Japanese quails reared in Abeokuta,Nigeria

Salmonellosis is a major bacterial disease causing huge economic losses in the poultry industry worldwide. This study was carried out to determine the period prevalence and antimicrobial susceptibility of Salmonella enterica in Japanese quails in Abeokuta, Nigeria. Four hundred cloacal swabs of quail birds were collected from 4 locations within Abeokuta. Salmonella was isolated from the samples using conventional methods for selective isolation of Salmonella and biochemical identification. Isolates were confirmed by polymerase chain reaction assays for the amplification and detection of Salmonella-associated virulence genes (invA and stn) using specific primers. Antimicrobial susceptibility testing was done using the Kirby-Bauer disk diffusion method. In all, Salmonella was isolated from 14 (3.5%) cloacal swabs. All 14 isolates possessed invA and stn genes. The Salmonella isolates showed resistance to tetracycline (100%), doxycycline (100%), ampicillin (100%), sulphamethoxazole (92.9%), nalidixic acid (85.8%), ceftazidime (78.6%), neomycin (64.3%), streptomycin (50%) and gentamycin (28.6%) but all the isolates were susceptible to ciprofloxacin. The isolates were resistant to at least three antimicrobials indicating multidrug resistance. The results concluded that Japanese quails harbour multidrug-resistant Salmonella which could be transmitted to humans through consumption of contaminated food or by direct and indirect contact with the carrier birds. Antimicrobial resistance could be due to overdependence on antimicrobials. Ciprofloxacin could be considered in the treatment of zoonotic Salmonellosis in humans.     

Antimicrobial resistance and recovery of Salmonella,Campylobacter, and Escherichia coli from chicken egg layer flocks in Canadian sentinel surveillance sites using 2 types of sample matrices

Eggs are important to the diet of Canadians. This product is one of the supply-managed commodities in Canada, but unlike other commodities, where food safety risks are extensively explored and reported, information on the prevalence of enteric organisms (e.g., Salmonella, Campylobacter) and antimicrobial resistance (AMR) in layers in Canada are limited. This study was conducted to determine the prevalence of select bacteria and the associated AMR patterns in layer flocks using 2 sample matrices. Farms were located within FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance sentinel sites (SS). Fecal samples (Ontario: ON_SS1a, ON_SS1b) and environmental sponge swabs (British Columbia: BC_SS2a) were collected. Salmonella prevalence was 29% and 8% in ON_SS1a and ON_SS1b, respectively, and 7% in BC_SS2a. S. Kentucky and S. Livingstone were the most frequently isolated serovars and no S. Enteritidis was detected. Campylobacter was not detected in the BC sponge swabs but was isolated from 89% and 53% of Ontario fecal samples (ON_SS1a and ON_SS1b, respectively). Seven C. jejuni from Ontario were ciprofloxacin-resistant. Escherichia coli prevalence was high in both sample types (98%). Overall, tetracycline resistance among E. coli ranged from 26% to 69%. Resistance to ceftiofur (n = 2 isolates) and gentamicin (n = 2) was relatively low. There were diverse resistance patterns (excludes susceptible isolates) observed among E. coli in Ontario (10 patterns) and British Columbia (14 patterns). This study revealed that fecal samples are more informative for farm-level monitoring of pathogen and AMR prevalence. Without further validation, sponge swabs are limited in their utility for Campylobacter detection and thus, for public health surveillance.     

Assessment of the aerobic faecal microflora in mink (Mustela vison Schreiber) with emphasis on Escherichia coli and Staphylococcus intermedius

The present study was undertaken to investigate the culturable aerobic faecal microflora of mink from newborn until adulthood with emphasis on the potential pathogens Escherichia coli and beta-haemolytic coagulase positive staphylococci. Rectal swabs were taken from 10 healthy dams and their offspring on seven mink farms throughout the production season and a semi-quantitative enumeration of total E. coli and haemolytic E. coli, beta-haemolytic streptococci, beta-haemolytic coagulase positive staphylococci, total lactic acid bacteria, and enterococci was carried out in all samples using selective and non-selective media.Aerobic bacteria were cultured from close to 100% of the samples throughout the survey. Prevalence of E. coli isolates varied between 70 and 90% of the samples throughout the survey with a small decline at the end of the study period. The highest bacterial counts were found among recently weaned kits or kits in the early growth period (P<0.0012). Lactic acid bacteria and enterococci were isolated from more than 90% of all samples, while beta-haemolytic staphylococci were isolated from 20 to 70% of the samples. While beta-haemolytic staphylococci were dominant from birth and during the nursing period, counts of staphylococci gradually decreased during the nursing period and were outnumbered by E. coli during the growth season.     

Antimicrobial resistance of Salmonella and generic Escherichia coli isolated from surface water samples used for recreation and a source of drinking water in southwestern Ontario,Canada

Antimicrobial resistance (AMR) in the aquatic environment represents an important means of introduction and dissemination of resistance genes, and presence of resistant pathogens in surface waters may pose a public health concern to recreational and drinking water users. The purpose of this study was to explore antimicrobial resistance patterns in water samples collected from the Grand River watershed (southwestern Ontario, Canada) to describe the composition, trends and potential risks of AMR in the aquatic environment. As part of FoodNet Canada and the Canadian Integrated Program for Antimicrobial Resistance Surveillance (CIPARS), stream water samples were collected bi‐weekly from sampling sites within the Grand River watershed in the Waterloo, Ontario sentinel site and tested for the presence and antimicrobial susceptibility of Salmonella spp. (2005–2013) and generic Escherichia coli (2012–2013). Of all samples tested, 16% of Salmonella and 22% of E. coli isolates were resistant to at least one antimicrobial, including three Salmonella isolates and two E. coli isolates that were resistant to Category I antimicrobials, which are classified as very high importance for the treatment of serious bacterial infections in humans. The greatest proportion of resistant E. coli isolates were observed from the river site upstream of the drinking water intake, while the greatest proportion of resistant Salmonella isolates were from sites upstream in the watershed, and at one recreational water site. Salmonella resistance trends remained fairly stable between 2007 and 2013, with the exception of streptomycin and tetracycline which increased in 2010 and 2013. Continued surveillance of antimicrobial resistance patterns and exploration of risk factor data will allow for a better understanding of resistance transmission in the aquatic environment.     

A direct plating method for monitoring the contamination of Listeria monocytogenes in silage.

Twenty-two silage samples were analyzed for the presence of L. monocytogenes using five Listeria selective plating media, with and without previous selective enrichment step. L. monocytogenes was recovered from 3 samples by both procedures, but direct plating allowed the quantification of Listeria population. Two of these positive samples were implicated in outbreaks of listeriosis in sheep; the L. monocytogenes population in these samples was about 10(6) cells/g. The L. monocytogenes population in the other positive sample was 10(3) cells/g. Direct isolation of L. monocytogenes was only possible from LPM, PALCAM and LSAMm media. MOX and LSM media were not selective enough to allow direct Listeria isolation. In our hands, LSAMm was the most suitable plating medium for the direct isolation and specific quantification of L. monocytogenes from silage employing a red blood cells overlay technique.     

Transmission of extended-spectrum cephalosporin-resistant Salmonella harboring a blaCMY-2-carrying IncA/C2 plasmid chromosomally integrated by ISEcp1 or IS26 in layer breeding chains in Japan

Dissemination of extended-spectrum cephalosporin (ESC)-resistant Salmonella is a public health concern in the egg production industry. ESC-resistant Salmonella often acquires the bla gene via insertion sequences (ISs). Therefore, this study aimed to assess antimicrobial resistance in Salmonella from Japanese layer breeding chains and egg processing chains, and determine the genetic profiles of IS-like elements in ESC-resistant Salmonella. Antimicrobial susceptibility testing was performed on 224 isolates from 49 facilities involving layer breeder farms, hatcheries, pullet-rearing farms, and layer farms in breeding chains along with egg processing chains. ESC-resistant Salmonella strains were whole-genome sequenced. Among them, 40 (17.9%) were resistant to at least streptomycin, tetracycline, ampicillin, chloramphenicol, cefpodoxime, nalidixic acid, ciprofloxacin, and/or kanamycin despite lacking resistance to azithromycin and meropenem. Moreover, 15 were ESC-resistant Salmonella harboring bla_CMY-2 (Salmonella enterica serovar Ohio, n=12; S. Braenderup, n=1; untypeable with O7:b:-, n=1) and bla_CTX-M-14 (S. Cerro, n=1). IncA/C₂ plasmids containing ISEcp1, IS26, and multiple antimicrobial resistance genes (including bla_CMY-2) were identified in S. Ohio isolates from pullet-rearing and layer farms belonging to the same company. Chromosomal integration of partial or whole IncA/C₂ plasmids was seen with two S. Ohio isolates via ISEcp1 or IS26, respectively. Antimicrobial resistance genes such as bla_CMY-2 might be transmitted among the upper and the lower levels of layer breeding chains via the replicon type IncA/C₂ plasmids containing ISEcp1 and IS26.     

Prevalence of Salmonella associated with chick mortality at hatching and their susceptibility to antimicrobial agents

The prevalence of Salmonella associated mortality at hatching was investigated in three hatcheries in Jos, central Nigeria. Their susceptibility to antimicrobial agents was also evaluated. S. Kentucky and S. Hadar were isolated. While half of the isolates were from internal organs, 26.7% came from meconial swabs of dead-in-shell embryos, 17.8% from intestinal samples and 4.4% from egg shells. S. Hadar is known to colonise only the gut and is classified as non-invasive, but in this study 82% were obtained from internal organs which suggests that infections with this serotype may also cause invasive disease. Antimicrobial susceptibility tests showed a high prevalence of antimicrobial resistance in the study area with complete resistance to gentamycin, enrofloxacin, nalidixic acid, tetracycline and streptomycin and substantial resistance to triple sulphur and ciprofloxacin. Six multiple resistance profiles were recorded with a high level of multiple resistance to quinolones. Quinolone resistance has implications for veterinary and human therapy as their misuse in poultry could lead to the emergence of resistant animal and zoonotic pathogens.     

Characteristics of ciprofloxacin and cephalosporin resistant Escherichia coli isolated from turkeys in Great Britain

1. A field study was performed to investigate the presence and characteristics of ciprofloxacin-resistant, extended spectrum β-lactamase (ESBL) and AmpC Escherichia coli from turkeys in Great Britain. E. coli were isolated from ～9000 boot swab samples from 27 different farms owned by four different companies. Between 1 and 14 visits were made to each farm (mean 3) at between 0 and 15?m intervals (mean?～?5?m).

2. CHROMagar ECC with and without ciprofloxacin or cephalosporin antibiotics was used as selective isolation media. Representative isolates with different phenotypes were tested for mutations in gyrA and for: qnrA, B, S, qepA and aac(6′)-Ib genes, for ESBL phenotype, the presence of bla _CTX-M genes and plasmid type, and for ampC genes_. Representative ciprofloxacin-resistant and CTX-M isolates were further tested for serotype and PFGE type. On ciprofloxacin selective media 55% of samples yielded ciprofloxacin resistant E. coli and of those further analysed, most had ciprofloxacin MICs >4 mg/l and mutations in gyrA.

3. For the different companies, the mean number of samples per farm with cefoxitin- or cefotaxime-resistant isolates ranged from 1·0% to 61·9% and 4·7% to 31·7% respectively. Cefotaxime-resistance was most commonly associated with an ESBL phenotype, a CTX-M-1 or CTX-M-14 sequence type and an I1-γ or K plasmid inc type. The mechanism of cefoxitin resistance was not determined for most isolates, but where determined it was bla _CMY-2.

4. PFGE and serotyping showed clonally-related isolates persisting over multiple visits suggesting both more prudent use of antibiotics and improved farm hygiene are needed to address the issue of antimicrobial resistance in isolates from turkeys.     

Effect of Recovery Medium on the Isolation of Campylobacter Jejuni Before and After Heat Treatment

The effect of various concentrations of polymyxin B and Colistin in Skirrow’s or Butzler’s type media, respectively, on the recovery rate of 13 strains of G. jejuni before and after heat treatment was studied. Prior to heating, a Butz-ler’s type medium containing 40 I.U. per ml Colistin was inhibitory for certain strains of C. jejuni. After heating at 48° G for 30 min, media containing 2.5 I.U. per ml polymyxin B or 10 I.U. per ml Colistin were not inhibitory for heat-injured cells. When the concentrations of polymyxin B were increased to 5.0 I.U. per ml or those of Colistin to 20 or 40 I.U. per ml in the selective media, the means of the differences of log cell counts between non-selective Brucella blood agar and the selective media was statistically significant (P < 0.05). The mean D-value of G. jejuni strains in Brucella broth at 48° G was 18.4±5.4 min.     

Occurrence and antimicrobial resistance of emergent Arcobacter spp. isolated from cattle and sheep in Iran

This study is conducted to determine the occurrence and antimicrobial resistance of Arcobacter spp. isolated from clinically healthy food animals. A total of 308 samples from cattle (200) and sheep (108) were collected from Shiraz slaughterhouse, southern Iran to investigate the presence of the important Arcobacter spp. using cultivation and Polymerase Chain Reaction (PCR) methods. Antimicrobial susceptibility of Arcobacter isolates was determined for 18 antibiotics using disk diffusion method. Among 308 samples, 27 (8.7%) and 44 (14.28%) were positive for the presence of Arcobacter species with cultivation and PCR procedures, respectively. The predominant species was A. butzleri in both cattle (58.33%) and sheep (55%). In addition, concurrent incidence of the species was observed in 25% of the positive samples. All Arcobacter isolates were resistant to rifampicin, vancomycin, ceftriaxone, trimethoprim and cephalothin. The isolates showed high susceptibility to tetracycline, oxytetracycline, erythromycin, ciprofloxacin, kanamycin, amikacin, gentamicin and enrofloxacin. No significant difference among cattle and sheep isolates in resistance pattern was observed. The results indicate that cattle and sheep are significant intestinal carriers for Arcobacter spp. Moreover, tetracycline and aminoglycosides showed great effects on Arcobacter species in antibiogram test and can be used for treatment of human Arcobacter infections.     

Effects of selective defoliation and height of defoliation on tiller dynamics and herbage yield of Themeda triandra in a semi-arid rangeland in Zimbabwe

Understanding the impacts of intensity and selection of grazing on the performance of grasses is important in the management of grazing areas. It is especially important in semi-arid environments where, apart from moisture, the levels at which grasses are utilized has a major influence on their persistence in the environment. The effects of selective defoliation and height of defoliation (5 cm and 10 cm stubble heights) on the performance of the grass, Themeda triandra, were investigated in a field experiment for two growing seasons. Performance was measured as tiller production, rate of production, tiller mortality, herbage yield and quality. Tiller production was greater (81 tillers per plant) under non-selective defoliation than under selective defoliation (49 tillers per plant) in the 1995/1996 season. Tiller mortality was higher (66.45%) under heavy selective defoliation than under nonselective defoliation (21.98%). Herbage yield, apart from the control treatment, was high (13.6 g per plant) under light non-selective defoliation. Heavy selective defoliation reduced the nutrient levels (e.g. levels of soluble carbohydrates under heavy selective defoliation were 6 g kg^?1 glucose compared to 20 g kg^?1 glucose under light non-selective defoliation).     

Antimicrobial resistance and prevalence of canine uropathogens at the Western College of Veterinary Medicine Veterinary Teaching Hospital, 2002–2007

Between January 2002 and June 2007, uropathogens were isolated from 473 of 1557 canine urine samples submitted to Prairie Diagnostic Services from the Western College of Veterinary Medicine Veterinary Teaching Hospital. Culture and susceptibility results were analyzed, retrospectively, to estimate the prevalence of common bacterial uropathogens in dogs with urinary tract infections and to identify changes in antimicrobial resistance. The most common pathogens identified were Escherichia coli, Staphylococcus intermedius, Enterococcus spp., and Proteus spp. Antimicrobial resistance increased during the study period, particularly among recurrent E. coli isolates. Using the formula to help select rational antimicrobial therapy (FRAT), bacterial isolates were most likely to be susceptible to gentamicin, fluoroquinolones, amoxicillin-clavulanic acid, and groups 4 and 5 (third generation) cephalosporins.     

A comparative study of the intestinal microbiota of healthy horses and those suffering from equine grass sickness

This study compares quantitatively the microbiota of the gastrointestinal tract of healthy horses with that of horses with equine grass sickness (EGS). Faecal and ileal samples were cultured quantitatively on selective and non-selective media. Confirmed anaerobes were identified to species level.Overall faecal counts gave a ratio of aerobes:anaerobes of approximately 1:1. However, the mean counts in healthy horses of 4.4x10(8) aerobes:3.7x10(8) anaerobes per gram wet weight were different from counts in EGS (means were 10-100-fold higher), with statistically significant differences for the anaerobes (p=0.04). There were 10-100-fold more anaerobic cocci in EGS samples compared to healthy controls. Most of the seven species of anaerobic cocci were found in both healthy horses and EGS. Differences in clostridia isolated between health and disease were notable: fourteen species were isolated from EGS cases, compared to only one (C. bifermentans) in controls. The mean faecal clostridial counts in chronic disease were higher than in controls (10-fold) and in acute EGS (100-fold). In contrast, mean counts for ileal samples from acute cases, showed a 10-fold increase for clostridia compared to 1000-fold reduction in chronic cases (compared to faecal counts). Results indicate an increase in the bacterial numbers in the GI tract of animals with EGS compared to the controls and clostridia are prominent in EGS. Whether the increase in clostridia is the cause of GI stasis or a consequence remains uncertain.