Farm level risk factors for fluoroquinolone resistance in E. coli and thermophilic Campylobacter spp. on poultry farms.

Data on husbandry practices, performance, disease and drug use were collected during a cross-sectional survey of 89 poultry meat farms in England and Wales to provide information on possible risk factors for the occurrence of fluoroquinolone (FQ)-resistant bacteria. Faeces samples were used to classify farms as "affected" or "not affected" by FQ-resistant (FQr) Escherichia coli or Campylobacter spp. Risk factor analysis identified the use of FQ on the farms as having by far the strongest association, among the factors considered, with the occurrence of FQr bacteria. Resistant E. coli and/or Campylobacter spp. were found on 86% of the farms with a history of FQ use. However, a substantial proportion of farms with no history of FQ use also yielded FQr organisms, suggesting that resistant bacteria may transfer between farms. Further analysis suggested that for Campylobacter spp., on-farm hygiene, cleaning and disinfection between batches of birds and wildlife control were of most significance. By contrast, for E. coli biosecurity from external contamination was of particular importance, although the modelling indicated that other factors were likely to be involved. Detailed studies on a small number of sites showed that FQr E. coli can survive routine cleaning and disinfection. It appears difficult to avoid the occurrence of resistant bacteria when FQ are used on a farm, but the present findings provide evidence to support recommendations to reduce the substantial risk of the incidental acquisition of such resistance by farms where FQ are not used.

A longitudinal observational study of Salmonella shedding patterns by commercial turkeys during rearing and fattening, showing limitations of some control measures.

1. The onset and progression of Salmonella infections was investigated in commercial turkey flocks from placement at 1 d old until slaughter in "brood and move" systems using a longitudinal observational approach based on faeces and environmental sampling with subsequent culture of Salmonella. 2. Persistent Salmonella Newport contamination was found within rearing houses and on their external concrete aprons after cleaning and disinfection between crops of heavily shedding young birds. 3. Salmonella shedding was often detected by 5 d of age and the frequency of positive samples peaked at 14-35 d. Thereafter Salmonella isolations declined, especially in the later (fattening) stages. Samples were still Salmonella-positive at low prevalence in half of the intensively sampled houses at slaughter age. 4. A number of management interventions to combat Salmonella infection of flocks, including sourcing policy, competitive exclusion cultures and cleaning and disinfection, were inadequate to prevent flock infection, although improved disinfection on one unit was associated with a delay in the onset of flock infection.

Efficacy of a live attenuated Escherichia coli O78:K80 vaccine in chickens and turkeys.

A candidate live vaccine for avian pathogenic Escherichia coli (APEC) was constructed from a virulent field APEC 078 strain by mutation of the aroA gene. The mutant was highly similar to the parent wild-type strain in respect of colony morphology, motility, growth in suspension, hemagglutination, Congo Red binding, HEp-2 cell adhesion, and the elaboration of surface antigens type 1 fimbriae and flagella, although production of curli fimbriae was reduced marginally. The mutant proved avirulent when inoculated into 1-day-old chicks by spray application and when presented again in the drinking water at 7 days of age. Chickens and turkeys vaccinated with an 078 aroA mutant were protected against a challenge at 6 wk of age by virulent APEC strains.

A critical review of Salmonella Typhimurium infection in laying hens.

Salmonella Typhimurium has been reported to contaminate egg production across the world, but where Salmonella Enteritidis is endemic it is this latter serovar that dominates egg-borne salmonellosis. However, Salmonella Typhimurium is a major food-borne pathogen so it is important to understand how it can impact the microbiological safety of eggs and what serovar-specific control strategies may be appropriate in the future as control over Salmonella Enteritidis continues to improve. To that end, the present review examines the published literature on Salmonella Typhimurium in laying hens and eggs, with particular reference to comparative studies examining different serovars. Experimentally Salmonella Enteritidis is more often isolated from egg contents and seems to adhere better to reproductive tract mucosa, whilst Salmonella Typhimurium appears to provoke a more intense tissue pathology and immune response, and flock infections are more transient. However, it is observed in many cases that the present body of evidence does not identify clear differences between specific behaviours of the serovars Typhimurium and Enteritidis, whether in laying hens, in their eggs, or in the laying environment. It is concluded that further long-term experimental and natural infection studies are needed in order to generate a clearer picture.

Investigations into Salmonella contamination in poultry feedmills in the United Kingdom.

AIMS: To investigate and compare commercial and farm-level milling operations in respect of the monitoring and control of Salmonella contamination. METHODS AND RESULTS: Four commercial feedmills and four on-farm poultry feed mixers were intensively sampled. Samples included dust and spillages and were cultured for Salmonella. Serovars in ingredients on farms were associated with wildlife and/or livestock, whereas those in commercial mill ingredients were associated with domestically produced cereals and imported vegetable protein. Endemic contamination of two commercial feedmills was reflected in isolates obtained from finished products and destination flocks. Renovation of equipment and chemical treatment of equipment and feed had not removed endemic strains, and previous routine monitoring in the commercial mills had not revealed the degree of contamination found in the present investigations. CONCLUSIONS: Ingredient contamination was diverse and reflected the sources and storage environments used by mills and farms, respectively. The use of dust and spillage samples showed a clear sensitivity advantage over the previously used monitoring methods in the feedmills. SIGNIFICANCE AND IMPACT OF THE STUDY: Monitoring for Salmonella contamination of commercial feedmills requires sensitive methods, such as those employed in this study. This is particularly important for endemic contamination.

Raw diets for dogs and cats: a review, with particular reference to microbiological hazards.

There is a recent trend to feed pet dogs and cats in Britain and other developed countries on raw meat and animal by-products using either commercial preparations or home recipes. This shift from heat-treated processed food has been driven by perceived health benefits to pets and a suspicion of industrially produced pet food. The diets of wild-living related species have been used as a rationale for raw feeding, but differences in biology and lifestyle impose limitations on such comparisons. Formal evidence does exist for claims by raw-feeding proponents of an altered intestinal microbiome and (subjectively) improved stool quality. However, there is currently neither robust evidence nor identified plausible mechanisms for many of the wide range of other claimed benefits. There are documented risks associated with raw feeding, principally malnutrition (inexpert formulation and testing of diets) and infection affecting pets and/or household members. Surveys in Europe and North America have consistently found Salmonella species in a proportion of samples, typically of fresh-frozen commercial diets. Another emerging issue concerns the risk of introducing antimicrobial-resistant bacteria. Raw pet food commonly exceeds hygiene thresholds for counts of Enterobacteriaceae. These bacteria often encode resistance to critically important antibiotics such as extended-spectrum cephalosporins, and raw-fed pets create an elevated risk of shedding such resistant bacteria. Other infectious organisms that may be of concern include Listeria, shiga toxigenic E scherichia coli , parasites such as Toxoplasma gondii and exotic agents such as the zoonotic livestock pathogen Brucella suis, recently identified in European Union and UK raw pet meat imported from Argentina.

A survey of fluoroquinolone resistance in Escherichia coli and thermophilic Campylobacter spp. on poultry and pig farms in Great Britain.

AIMS: To estimate the proportions of farms on which broilers, turkeys and pigs were shedding fluoroquinolone (FQ)-resistant Escherichia coli or Campylobacter spp. near to slaughter. METHODS AND RESULTS: Freshly voided faeces were collected on 89 poultry and 108 pig farms and cultured with media containing 1.0 mg l(-1) ciprofloxacin. Studies demonstrated the specificity of this sensitive method, and both poultry and pig sampling yielded FQ-resistant E. coli on 60% of farms. FQ-resistant Campylobacter spp. were found on around 22% of poultry and 75% of pig farms. The majority of resistant isolates of Campylobacter (89%) and E. coli (96%) tested had minimum inhibitory concentrations for ciprofloxacin of > or =8 mg l(-1). The proportion of resistant E. coli and Campylobacter organisms within samples varied widely. CONCLUSIONS: FQ resistance is commonly present among two enteric bacterial genera prevalent on pig and poultry farms, although the low proportion of resistant organisms in many cases requires a sensitive detection technique. SIGNIFICANCE AND IMPACT OF THE STUDY: FQ-resistant bacteria with zoonotic potential appear to be present on a high proportion of UK pig and poultry farms. The risk this poses to consumers relative to other causes of FQ-resistant human infections remains to be clarified.

Survey of the prevalence of Salmonella species on commercial laying farms in the United Kingdom.

A survey of salmonella infection on 454 commercial layer flock holdings in the uk was carried out between October 2004 and September 2005. Fifty-four (11.7 per cent, 95 per cent confidence interval 9.3 to 14.0 per cent) were salmonella positive. The most common serovar identified was Salmonella Enteritidis at a prevalence of 5.8 per cent, and 70 per cent of these isolates were phage types 4, 6, 7 and 35. Salmonella Typhimurium was the second most prevalent serovar, found in 1.8 per cent of the farms. Of the three other serovars given top priority by the eu because of their public health significance, Salmonella Virchow and Salmonella Infantis were each isolated from one holding, but Salmonella Hadar was not isolated from any of the holdings. Analysis of antimicrobial resistance patterns revealed that over 76 per cent of the isolates were sensitive to all of the 16 drugs tested, and all the isolates were sensitive to ciprofloxacin, gentamicin, ceftazidime, apramycin, amikacin, amoxicillin/clavulanic acid, neomycin and cefotaxime.

Salmonella Vaccination in Pigs: A Review.

The control of Salmonella enterica in pig production is necessary for both public and animal health. The persistent and frequently asymptomatic nature of porcine Salmonella infection and the organism's abilities to colonize other animal species and to survive in the environment mean that effective control generally requires multiple measures. Vaccination is one such measure, and the present review considers its role and its future, drawing on studies in pigs from the 1950s to the present day. Once established in the body as an intracellular infectious agent, Salmonella can evade humoral immunity, which goes some way to explaining the often disappointing performance of inactivated Salmonella vaccines. More recent approaches, using mucosal presentation of antigens, live vaccines and adjuvants to enhance cell-mediated immunity, have met with more success. Vaccination strategies that involve stimulating both passive immunity from the dam plus active immunity in offspring appear to be most efficacious, although either approach alone can yield significant control of Salmonella. Problems that remain include relatively poor control of Salmonella serovars that are dissimilar to the vaccine antigen mix, and difficulties in measuring and predicting the performance of candidate vaccines in ways that are highly relevant to their likely use in commercial production.

Potential role of multiple rectal biopsies in the diagnosis of equine grass sickness.

Two samples were taken postmortem from the rectum of each of 14 horses with grass sickness and 10 control horses, and four sections stained with haematoxylin and eosin were examined. By using as a criterion of grass sickness the presence of three chromatolytic neurons, 10 of the 14 cases were positive and none of the control horses was positive, giving a sensitivity of 71 per cent and a specificity of 100 per cent. No other histological features appeared to be of diagnostic value, and staining for Nissl substance in neurons with cresyl fast violet and methyl green-pyronin did not improve the sensitivity of the test.

Use of an attenuated live Salmonella Typhimurium vaccine on three breeding pig units: A longitudinal observational field study.

The study examined the effects of a licensed live Salmonella Typhimurium vaccine, administered to sows and gilts on three commercial pig units experiencing clinical salmonellosis associated with S. Typhimurium or its monophasic variant. After vaccination, clinical salmonellosis resolved and shedding of S. Typhimurium declined markedly and persistently on all breeding or breeding-finishing units, during the one- to two-year monitoring period. On two finishing units supplied in part by one of the vaccinated herds, pigs from the vaccinated herd were less likely to shed Salmonella than those from non-vaccinating herds, and Salmonella counts in faeces were also lower from the vaccine-linked animals. Non-Typhimurium Salmonella serovars were isolated typically in fewer than 10% of samples, and showed no clear temporal changes in frequency. Vaccination of dams alone with S. Typhimurium was associated with reduced shedding of closely-related serovars among all age groups in this commercial setting.

Attaching-effacing bacteria in animals.

Enteric bacteria with a demonstrable or potential ability to form attaching-effacing lesions, so-called attaching-effacing (AE) bacteria, have been found in the intestinal tracts of a wide variety of warm-blooded animal species, including man. In some host species, for example cattle, pigs, rabbits and human beings, attaching-effacing Escherichia coli (AEEC) have an established role as enteropathogens. In other host species, AE bacteria are of less certain significance. With continuing advances in the detection and typing of AE strains, the importance of these bacteria for many hosts is likely to become clearer. The pathogenic effects of AE bacteria result from adhesion to the intestinal mucosa by a variety of mechanisms, culminating in the formation of the characteristic intimate adhesion of the AE lesion. The ability to induce AE lesions is mediated by the co-ordinated expression of some 40 bacterial genes organized within a so-called pathogenicity island, known as the "Locus for Enterocyte Effacement". It is also believed that the production of bacterial toxins, principally Vero toxins, is a significant virulence factor for some AEEC strains. Recent areas of research into AE bacteria include: the use of Citrobacter rodentium to model human AEEC disease; quorum-sensing mechanisms used by AEEC to modulate virulence gene expression; and the potential role of adhesion in the persistent colonization of the intestine by AE bacteria. This review of AE bacteria covers their molecular biology, their occurrence in various animal species, and the diagnosis, pathology and clinical aspects of animal diseases with which they are associated. Reference is made to human pathogens where appropriate. The focus is mainly on natural colonization and disease, but complementary experimental data are also included.

Attaching-effacing lesions associated with Escherichia coli O157:H7 and other bacteria in experimentally infected conventional neonatal goats.

Four conventionally reared goats aged 6 days were inoculated orally with approximately 10(10) colony-forming units (cfu) of a non-verotoxigenic strain of Escherichia coli O157:H7. All remained clinically normal. Tissues were sampled under terminal anaesthesia at 24 (two animals), 48 and 72 h post-inoculation (hpi). E. coli O157:H7 was cultured from the ileum, caecum, colon and rectum of all animals, but the number of bacteria recovered at these sites varied between animals. Attaching-effacing (AE) lesions associated with O157 organisms, as confirmed by immunolabelling, were observed in the ileum of one of the two animals examined at 24 hpi, and in the ileum, caecum and proximal colon of an animal examined at 72 hpi. E. coli O157 organisms were detected at > or =10(5) cfu/g of tissue at these sites. In addition, AE lesions associated with unidentified bacteria were observed at various sites in the large bowel of the same animals. Lesions containing both E. coli O157 and unidentified bacteria (non-O157) were not observed. Non-O157 AE lesions were also observed in the large bowel of one of two uninoculated control animals. This indicated that three (one control and two inoculated) animals were colonized with an unidentified AE organism before the commencement of the experiment. The O157-associated AE lesions were observed only in animals colonized by non-O157 AE organisms and this raises questions about individual host susceptibility to AE lesions and whether non-O157 AE organisms influence colonization by E. coli O157.

Naturally acquired attaching and effacing Escherichia coli in sheep.

In a series of experiments involving the inoculation of sheep with Escherichia coli O157:H7, and subsequent detailed histopathological examination of the intestinal mucosa, attaching-effacing (AE) lesions formed by elements of the natural flora were observed in 18% of animals. These incidental AE lesions typically were small and sparse, and were not associated with clinical disease. It was possible to identify further some of the lesional bacteria, revealing that E. coli O115 had formed lesions in one of the seven affected animals, and similarly E. coli O26 had formed some of the lesions in another. As AE strains, source flocks, housing and feed sources were diverse, a common source of lesion-forming bacteria appears to be unlikely. It is postulated that subclinical AE lesions are a mechanism of persistence of AE bacteria in sheep.

A survey of equine abortion, stillbirth and neonatal death in the UK from 1988 to 1997.

REASONS FOR PERFORMING STUDY: A detailed review of laboratory records for equine abortion is fundamental in establishing current disease trends and suggesting problems important for further research. OBJECTIVES: To review the causes of abortion and neonatal death in equine diagnostic submissions to the Animal Health Trust over a 10 year period. METHODS: The diagnoses in 1252 equine fetuses and neonatal foals were reviewed and analysed into categories. RESULTS: Problems associated with the umbilical cord, comprising umbilical cord torsion and the long cord/cervical pole ischaemia disorder, were the most common diagnoses (38.8%: 35.7% umbilical cord torsion and 3.1% long cord/cervical pole ischaemia disorder). Other noninfective causes of abortion or neonatal death included twinning (6.0%), intrapartum stillbirth (13.7%) and placentitis, associated with infection (9.8%). E. coli and Streptococcus zooepidemicus were the most common bacteria isolated. Neonatal infections not associated with placentitis accounted for 3.2% of incidents; and infections with EHV-1 or EHV-4 for 6.5%. CONCLUSIONS: Definitive diagnosis of equine abortion is possible in the majority of cases where the whole fetus and placenta are submitted for examination. POTENTIAL RELEVANCE: Given the high incidence of umbilical cord torsion and related problems as causes of abortion in UK broodmares, more research on factors determining umbilical cord length and risk of torsion is essential.

The diagnosis of lameness associated with distal limb pathology in a horse: a comparison of radiography, computed tomography and magnetic resonance imaging.

A cadaver limb from an eight-year-old horse with right forelimb lameness that was relieved with an intra-articular distal interphalangeal joint block was imaged with radiographs, spiral computed tomography (CT) and magnetic resonance imaging (MRI). Spiral CT demonstrated several lucencies within the deep digital flexor tendon immediately proximal to the navicular bone. On MRI these areas had increased signal and there was enlargement of the tendon at this site. Effusion in the proximal interphalangeal joint and navicular bursa and thinning of the fibrocartilage of the navicular bone were also observed on MRI images. These changes were not detected on radiographs. Histopathology confirmed that there were focal areas of collagen necrosis within the deep digital flexor tendon with thinning and degenerative changes in the fibrocartilage of the navicular bone.

Salmonella contamination of cereal ingredients for animal feeds.

Cereal ingredients for animal feedstuffs may become contaminated by Salmonella on their farms of origin. This is often concentrated in multiple foci, owing to contamination by rodents and other wildlife which may be missed by routine sampling, and may involve serovars of particular public health significance, such as Salmonella Typhimurium (STM). The study examined such contamination in domestically-produced cereal ingredients in the United Kingdom. Cereal-producing farms with associated cattle or pig enterprises (43) and feedmills (6) were investigated, following the isolation of STM from their premises (feedmills) or STM DT104 from their livestock (farms) by routine surveillance. Cereal samples from feedmills yielded two STM isolates from the same premises, of the same phage types as were isolated from wild bird faeces at ingredient intake and product loading areas. Farm investigations identified numerous Salmonella serovars, including STM, on grain harvesting and handling equipment, in grain storage areas, and in wildlife samples. Mice were removed from one pig farm and shed Salmonella Derby and Salmonella Bovismorbificans for 10 months afterwards. Grain stores more than one kilometre away from livestock areas were rarely found to be contaminated with STM. The principal issues with Salmonella contamination of cereals appeared to be the use of livestock areas as temporary grain stores on cattle farms, and access to stored grain by wildlife and domestic animals.

A comparison between longitudinal shedding patterns of Salmonella Typhimurium and Salmonella Dublin on dairy farms.

Salmonella in cattle herds may behave as epidemic or endemic infections. An intensive longitudinal sampling study across all management groups and ages on six dairy farms in the UK was used to examine patterns of Salmonella shedding, following the prior identification of either Salmonella Dublin (SD) (three farms) or Salmonella Typhimurium (ST) (three farms) on the premises in the context of clinical salmonellosis. Individual faeces, pooled faeces and environmental samples (total 5711 samples), taken approximately every six weeks for 15-24 weeks, were cultured for Salmonella. SD was detected at low frequency (on any visit, 0.5-18.3 per cent of samples positive) and most consistently in calves. By contrast, ST was isolated at higher frequency (on any visit, 6.8-75 per cent of samples positive), and in higher numbers, up to 10(7) cfu/g faeces. Significantly more samples from calves were positive for ST than were positive for SD (50.6 per cent v 3.1 per cent; P < 0.001), which was also true for milking cows (46.3 per cent v 4.4 per cent; P < 0.001). The differences could help to explain the different patterns of bovine infection classically associated with these two serovars in the UK. No consistent effect upon shedding was seen among the ST-infected herds following vaccination.

Escherichia coli O115 forms fewer attaching and effacing lesions in the ovine colon in the presence of E. coli O157:H7.

Escherichia coli O115 has been isolated from healthy sheep and was shown to be associated with attaching-effacing (AE) lesions in the large intestine. Following previous observations of interactions between E. coli O157 and O26, the aim of the present study was to assess what influence an O115 AE E. coli (AEEC) would have on E. coli O157 colonisation in vitro and in vivo. We report that E. coli O115- and O157-associated AE lesions were observed on HEp-2 cells and on the mucosa of ligated ovine spiral colon. In single strain inoculum, E. coli O115 associated intimately with HEp-2 cells and the spiral colon in greater numbers than E. coli O157:H7. However, in mixed inoculum studies, the number of E. coli O115 AE lesions was significantly reduced suggesting negative interference by E. coli O157. Use of the ligated colon model in the present work has allowed in vitro observations to be extended and confirmed whilst using a minimum of experimental animals. The findings support a hypothesis that some AEEC can inhibit adhesion of other AEEC in vivo. The mechanisms involved may prove to be of utility in the control of AE pathovars.

Producing Salmonella-free pigs: a review focusing on interventions at weaning.

Salmonella infection in pig production is typically endemic and largely asymptomatic. It is a cause of substantial concern among food safety bodies, prompting voluntary and legislative responses aimed at monitoring and reducing the number of Salmonella-infected animals entering the human food chain. Elimination of the problem at an early stage of production is highly desirable, and to this end the present review examines published evidence on the carriage of Salmonella by piglets before and after weaning, as well as evidence on the dynamics of Salmonella infection in the weaner and grower stages of pig production, the effects of maternal immunity, and risk factors for Salmonella excretion after weaning. Various interventions to reduce or eliminate Salmonella infection in young pigs have been tried, such as vaccination, competitive exclusion, treatments in feed and water, antibiotic administration, disinfection of animals, and segregated weaning to clean accommodation. The evidence on the effectiveness of these is considered, and the last is examined in some detail, as it appears currently to offer the best chance of eliminating Salmonella from growing stock.