Iain J. McKendrick

Lymphoid Follicle-Dense Mucosa at the Terminal Rectum Is the Principal Site of Colonization of Enterohemorrhagic Escherichia coli O157:H7 in the Bovine Host

ABSTRACT Escherichia coli O157:H7 causes bloody diarrhea and potentially fatal systemic sequelae in humans. Cattle are most frequently identified as the primary source of infection, and E. coli O157:H7 generally colonizes the gastrointestinal tracts of cattle without causing disease. In this study, persistence and tropism were assessed for four different E. coli O157:H7 strains. Experimentally infected calves shed the organism for at least 14 days prior to necropsy. For the majority of these animals, as well as for a naturally colonized animal obtained from a commercial beef farm, the highest numbers of E. coli O157:H7 were found in the feces, with negative or significantly lower levels detected in lumen contents taken from the gastrointestinal tract. Detailed examination demonstrated that in these individuals the majority of tissue-associated bacteria were adherent to mucosal epithelium within a defined region extending up to 5 cm proximally from the recto-anal junction. The tissue targeted by E. coli O157:H7 was characterized by a high density of lymphoid follicles. Microcolonies of the bacterium were readily detected on the epithelium of this region by immunofluorescence microscopy. As a consequence of this specific distribution, E. coli O157:H7 was present predominately on the surface of the fecal stool. In contrast, other E. coli serotypes were present at consistent levels throughout the large intestine and were equally distributed in the stool. This is a novel tropism that may enhance dissemination both between animals and from animals to humans. The accessibility of this site may facilitate simple intervention strategies.

Paratuberculosis Infection of Nonruminant Wildlife in Scotland

ABSTRACT Recent reports of natural paratuberculosis (or Johnes disease) in rabbits, foxes, and stoats has focused debate on the presence and importance of wildlife reservoirs in the epidemiology of this disease. This paper describes an extensive study investigating 18 nonruminant wildlife species for evidence of paratuberculosis. Using both culture and histopathological analysis, fox, stoat, weasel, crow, rook, jackdaw, rat, wood mouse, hare, and badger were found to harborMycobacterium avium subsp. paratuberculosis, the causative organism of paratuberculosis, suggesting that the epidemiology of this disease is more complex than previously realized.

Rectal Carriage of Enterohemorrhagic Escherichia coli O157 in Slaughtered Cattle

ABSTRACT Escherichia coli O157:H7 is an important cause of diarrhea, hemorrhagic colitis, and potentially fatal human illness. Cattle are considered a primary reservoir of infection, and recent experimental evidence has indicated that the terminal rectum is the principal site of bacterial carriage. To test this finding in naturally colonized animals, intact rectum samples from 267 cattle in 24 separate lots were obtained immediately after slaughter, and fecal material and mucosal surfaces were cultured for E. coli O157 by direct and enrichment methods. Two locations, 1 and 15 cm proximal to the recto-anal junction, were tested. In total, 35 animals were positive for E. coli O157 at at least one of the sites and 232 animals were negative as determined by all tests. The frequency of isolation and the numbers of E. coli O157 cells were higher at the site closer to the recto-anal junction, confirming our previous experimental findings. We defined low- and high-level carriers as animals with E. coli O157 levels of <1 × 103 CFU g−1 or <1 × 103 CFU ml−1 and animals with E. coli O157 levels of ≥1 × 103 CFU g−1 or ≥1 × 103 CFU ml−1 in feces or tissues, respectively. High-level carriage was detected in 3.7% of the animals (95% confidence interval, 1.8 to 6.8%), and carriage on the mucosal surface of the terminal rectum was associated with high-level fecal excretion. In summary, our results support previous work demonstrating that the mucosal epithelium in the bovine terminal rectum is an important site for E. coli O157 carriage in cattle. The data also support the hypothesis that high-level fecal shedding (≥1 × 103 CFU g of feces−1) of enterohemorrhagic E. coli O157 results from colonization of this site.

Super-shedding cattle and the transmission dynamics of Escherichia coli O157

The prevalence of Escherichia coli O157 displays striking variability across the Scottish cattle population. On 78% of farms, in a cross-sectional survey of 952, no shedding of E. coli O157 was detected, but on a small proportion, approximately 2%, very high prevalences of infection were found (with 90-100% of pats sampled being positive). We ask whether this variation arises from the inherent stochasticity in transmission dynamics or whether it is a signature of underlying heterogeneities in the cattle population. A novel approach is taken whereby the cross-sectional data are viewed as providing independent snapshots of a dynamic process. Using maximum-likelihood methods to fit time-dependent epidemiological models to the data we obtain estimates for the rates of immigration and transmission of E. coli O157 infection - parameters which have not been previously quantified in the literature. A comparison of alternative model fits reveals that the variation in the prevalence data is best explained when a proportion of the cattle are assumed to transmit infection at much higher levels than the rest - the so-called super-shedders. Analysis of a second dataset, comprising samples taken from 32 farms at monthly intervals over a period of 1 year, additionally yields an estimate for the rate of recovery from infection. The pattern of prevalence displayed in the second dataset also strongly supports the super-shedder hypothesis.

Risk Factors for the Presence of High-Level Shedders of Escherichia coli O157 on Scottish Farms

ABSTRACT Escherichia coli O157 infections are the cause of sporadic or epidemic cases of often bloody diarrhea that can progress to hemolytic uremic syndrome (HUS), a systematic microvascular syndrome with predominately renal and neurological complications. HUS is responsible for most deaths associated with E. coli O157 infection. From March 2002 to February 2004, approximately 13,000 fecal pat samples from 481 farms with finishing/store cattle throughout Scotland were examined for the presence of E. coli O157. A total of 441 fecal pats from 91 farms tested positive for E. coli O157. From the positive samples, a point estimate for high-level shedders was identified using mixture distribution analysis on counts of E. coli O157. Models were developed based on the confidence interval surrounding this point estimate (high-level shedder, greater than 103 or greater than 104 CFU g−1 feces). The mean prevalence on high-level-shedding farms was higher than that on low-level-shedding farms. The presence of a high-level shedder on a farm was found to be associated with a high proportion of low-level shedding, consistent with the possibility of a higher level of transmission. Analysis of risk factors associated with the presence of a high-level shedder on a farm suggested the importance of the pathogen and individual host rather than the farm environment. The proportion of high-level shedders of phage 21/28 was higher than expected by chance. Management-related risk factors that were identified included the type of cattle (female breeding cattle) and cattle stress (movement and weaning), as opposed to environmental factors, such as water supply and feed.

Absence of urinary opioid peptides in children with Autism

Objective: It has been claimed for a number of years that the urine of children with autism contains exogenously derived opioid peptides. This finding is said to reflect a disturbance in the integrity of the gut epithelium, act as a diagnostic marker for autism and predict treatment response to a diet excluding gluten and casein. The aim of the present study was to determine whether exogenous or endogenous peptides were present in the urine of children with autism or of control children. Design: Case-control study Setting: Cases were recruited from two tertiary referral centres specialising in autistic spectrum disorders, while controls were recruited from mainstream primary and secondary schools in the same geographical area. Participants: 65 boys with autism, mean age 7.4 years (range 5–11) and 158 control boys, mean age 7.8 years (range 4.2–11). Investigations: Urine samples were examined by high pressure liquid chromatography (HPLC) and matrix assisted laser desorbtion ionisation-time of flight mass spectrometry (MALDI-TOF MS) for the presence of a number of putative opioid peptides. Outcomes: There were no significant differences between the HPLC urinary profiles of the children affected by autism and the typically developing controls. In those cases where HPLC showed peaks in the locations at which opioid peptides might be expected to be found, MALDI-TOF established that these peaks did not, in fact, represent opioid peptides. Conclusions: Given the lack of evidence for any opioid peptiduria in children with autism, opioid peptides can neither serve as a biomedical marker for autism nor be employed to predict or monitor response to a casein- and gluten-free diet.

Successful immunization against a parasitic nematode by vaccination with recombinant proteins

Infection of humans and livestock with parasitic nematodes can have devastating effects on health and production, affecting food security in both developed and developing regions. Despite decades of research, the development of recombinant sub-unit vaccines against these pathogens has been largely unsuccessful. We have developed a strategy to identify protective antigens from Teladorsagia circumcincta, the major pathogen causing parasitic gastroenteritis in small ruminants in temperate regions, by studying IgA responses directed at proteins specific to post-infective larvae. Antigens were also selected on the basis of their potential immunomodulatory role at the host/parasite interface. Recombinant versions of eight molecules identified by immunoproteomics, homology with vaccine candidates in other nematodes and/or with potential immunoregulatory activities, were therefore administered to sheep in a single vaccine formulation. The vaccine was administered three times with Quil A adjuvant and the animals subsequently subjected to a repeated challenge infection designed to mimic field conditions. Levels of protection in the vaccinates were compared to those obtained in sheep administered with Quil A alone. The trial was performed on two occasions. In both trials, vaccinates had significantly lower mean fecal worm egg counts (FWECs) over the sampling period, with a mean reduction in egg output of 70% (Trial 1) and 58% (Trial 2). During the period of peak worm egg shedding, vaccinates shed 92% and 73% fewer eggs than did controls in Trials 1 and 2, respectively. At post mortem, vaccinates had 75% (Trial 1) and 56% (Trial 2) lower adult nematode burdens than the controls. These levels of protection are the highest observed in any system using a nematode recombinant sub-unit vaccine in the definitive ruminant host and indicate that control of parasitic helminths via vaccination with recombinant subunit vaccine cocktails is indeed an alternative option in the face of multi-drug resistance.

Using a Bayesian belief network to aid differential diagnosis of tropical bovine diseases.

The examination of presenting signs has always played an important role in the diagnosis of diseases in animal populations. In the case of diseases of tropical cattle, such expertise is often scarce and confined to those experts with many years of experience. To capture, conserve and disseminate such valuable expert knowledge remains a key challenge to the application of knowledge-based systems in veterinary medicine. In this communication, we explore the use of a Bayesian belief network to quantify expert opinion with a view to estimating the likelihood of various diseases in the presence and absence of certain signs. Information was elicited from a panel of 44 experienced veterinarians to provide the response matrix of 27 signs associated with 20 commonly occurring diseases in sub-Saharan cattle. Using this prior information, estimates of the probability of certain signs occurring with each disease were calculated from which the Bayesian belief network was able to propagate the posterior probability of each of the diseases based on the observed signs. The method as an aid in making diagnosis is discussed. It is recognised that such an approach is but one strand in the process of arriving at a diagnosis. For ease of use and accessibility, the approach has been converted into the software program CaDDiS (Cattle Disease Diagnosis System) which is available for consultation on the World Wide Web.

Comparison of three alternative methods for analysis of equine Faecal Egg Count Reduction Test data.

The Faecal Egg Count Reduction Test (FECRT) is the most widely used method of assessing the efficacy of anthelmintics, and is the only in vivo technique currently approved for use with horses. Equine Faecal Egg Count (FEC) data are frequently characterised by a low mean, high variability, small sample size and frequent zero count observations. Accurate analysis of the data therefore depends on the use of an appropriate statistical technique. Analyses of simulated FECRT data by methods based on calculation of the empirical mean and variance, non-parametric bootstrapping, and Markov chain Monte Carlo (MCMC) are compared. The MCMC method consistently outperformed the other methods, independently of the distribution from which the data were generated. Bootstrapping produced notional 95% confidence intervals containing the true parameter as little as 40% of the time with sample sizes of less than 50. Analysis of equine FECRT data yielded inconclusive results in 53 of 63 (84%) datasets, suggesting that the routine use of prior sample size calculations should be adopted to ensure sufficient data are collected. The authors conclude that computationally intensive parametric methods such as MCMC be used for analysis of FECRT data with sample sizes of less than 50, in order to avoid erroneous inference about the true efficacy of anthelmintics in the field.

Risk factors for Johne's disease in Scotland - the results of a survey of farmers

The reported incidence of Johnes disease has been increasing in the east of Scotland since 1993. A postal questionnaire survey was sent to 127 farms to identify potential risk factors for Johnes disease in relation to wildlife and farm management practices, and 86 returns were obtained. Of 22 farms which had been assumed to be free of the disease, on the basis of information held by local veterinary centres, seven (32 per cent) reported cases of Johnes disease in the 1990s, indicating that the disease is under-reported. Logistic regression analyses showed that eight of 63 potentially explanatory variables were significant at the 5 per cent level in affecting the likelihood of farms reporting Johnes disease. Of these, large numbers of livestock and rabbits, and access of wildlife to feed stores were the clearest and most consistent risk factors associated with the disease. The application of manure to grazing pasture, the type of water supply for the cattle and the numbers of crows were also related to the presence of Johnes disease but the nature of these relationships was less clear. Only 38 per cent of the farms reported taking any control measures to combat Johnes disease, but three of the control measures were relevant to risk factors identified as significant by the survey, namely maintaining a clean water supply, controlling rabbits and not spreading manure on to grazing pasture.