Toxigenic Corynebacterium ulcerans associated with upper respiratory infections in cats and dogs.

OBJECTIVES: To describe infection in companion animals with the zoonotic pathogen Corynebacterium ulcerans and to determine its prevalence in clinically-affected and healthy animals. MATERIALS AND METHODS: The clinical presentation and treatment of three cases of C. ulcerans infection is described. Two studies to determine C. ulcerans prevalence rates were undertaken: (a) a prospective study of nasal samples from healthy animals, 479 dogs and 72 cats; (b) a retrospective analysis of records of nasal samples collected over a 10-year period from 189 dogs and 64 cats affected by respiratory signs. RESULTS: Toxigenic C. ulcerans was isolated from four cats with nasal discharge while concurrent C. ulcerans and mecC methicillin-resistant S. aureus infection was detected in a dog suffering from chronic nasal discharge. Clinical features were not distinctive and all cases recovered following antimicrobial treatment. Multilocus sequence typing supported a common source for isolates from the shelter cats. Carriage rates of C. ulcerans in healthy animals were 0.42% (2/479) in dogs and 0.00% (0/72) in cats whereas in animals with signs of upper respiratory tract infection prevalence rates were 0.53% (1/189) in dogs and 6.25% (4/64) in cats. CLINICAL SIGNIFICANCE: Clinicians should be aware that dogs and cats can be infected with (or carriers of) toxigenic C. ulcerans Considering the potential zoonotic risk, assistance from medical and public health colleagues should be sought in confirmed cases.

Salmonella in breeding pigs: Shedding pattern, transmission of infection and the role of environmental contamination in Irish commercial farrow-to-finish herds.

This study aimed to provide new insights into the epidemiology of Salmonella in pig production, focusing on potential shedding patterns in breeding pigs throughout a full production cycle and the risk of transmission of infection from the sow to her offspring. A longitudinal study was conducted on five farrow-to-finish commercial pig farms. In each herd, shedding of Salmonella in faeces was monitored in breeders through service, gestation and lactation. Swabs of the farrowing room floor and pools of faeces from piglets were collected on two occasions during lactation. Environmental pen swabs were also taken in the weaning and finisher houses. Salmonella isolates were serotyped, tested for antimicrobial resistance (AMR) and typed by Multiple-Locus Variable number tandem repeat Analysis (MLVA). Shedding by breeding pigs was low in all stages of the production cycle; 5% of sows shed at service, the production stage with highest risk of shedding (p < .01), 1.6% shed during gestation and 2.5% after farrowing. Salmonella was detected in 4% of piglet faecal pools in the second week post-farrowing and 5% in the fourth week. Serotyping and AMR profiles of Salmonella isolates revealed that strains in sows and gilts were mostly different from strains isolated in weaner and finisher facilities. MLVA typing confirmed that the source of infection in piglets was in most instances the contaminated environment rather than their dam. Based on the typing results, it appears that sows do not pose a major risk in the maintenance and transmission of Salmonella to their progeny but instead the contaminated pen environment is more significant in the perpetuation of the organism on farm.

Investigation of in-feed organic acids as a low cost strategy to combat Salmonella in grower pigs.

Salmonella carriage in pigs is a significant food safety issue. Dietary supplementation with organic acids has previously been shown to reduce shedding and transmission of Salmonella. Therefore, this study aimed to examine the effect of three commercially available organic acid-based products on Salmonella levels in grower pigs, using a model of experimental infection that closely mimics natural exposure to the organism. Seven week old trial pigs (n=40) with a mean weight of 14.7kg were placed in one of four pens with 10 pigs/pen. Pens had previously been contaminated with Salmonella Typhimurium 4,[5],12;i;- via seeder pigs. Trial pigs received one of four diets for 28days: 1, control diet; 2, sodium butyrate supplemented diet; 3, benzoic acid supplemented diet and 4, formic-citric acid supplemented diet. A further 10 pigs were placed in a Salmonella-free pen receiving the control diet. Pigs were weighed and blood sampled on days 0 and 28. Faeces was collected on day 0, 2, 3, 5, 7, 14, 21 and 28 and examined for Salmonella. On day 28, 5 pigs/group were euthanised and ileocaecal lymph nodes (ILN) and caecal contents sampled for culture. The remaining 5 pigs/pen were then fed the control diet and faeces were collected on days 35 and 42. On day 42 pigs were euthanised and ILN and caecal contents tested for Salmonella levels. The trial was repeated once. Within the first two days of exposure to the contaminated environment, 96% (77/80) of pigs became infected. Most pigs shed Salmonella at levels of between 100-103 CFU/g faeces for at least 7days post-exposure. A significant reduction in Salmonella faecal concentration was observed after supplementation with sodium butyrate (p=0.001) and a formic citric acid blend (p<0.0001). Average daily weight gain (ADWG) was significantly increased in all groups fed the supplemented feed when compared to the positive control group. The use of sodium butyrate or a blend of formic and citric acid in feed could be considered a cost-effective control measure to reduce Salmonella faecal shedding and improve ADWG in Salmonella infected herds.

Antimicrobial Resistance of Faecal Escherichia coli Isolates from Pig Farms with Different Durations of In-feed Antimicrobial Use.

Antimicrobial use and resistance in animal and food production are of concern to public health. The primary aims of this study were to determine the frequency of resistance to 12 antimicrobials in Escherichia coli isolates from 39 pig farms and to identify patterns of antimicrobial use on these farms. Further aims were to determine whether a categorization of farms based on the duration of in-feed antimicrobial use (long-term versus short-term) could predict the occurrence of resistance on these farms and to identify the usage of specific antimicrobial drugs associated with the occurrence of resistance. Escherichia coli were isolated from all production stages on these farms; susceptibility testing was carried out against a panel of antimicrobials. Antimicrobial prescribing data were collected, and farms were categorized as long term or short term based on these. Resistance frequencies and antimicrobial use were tabulated. Logistic regression models of resistance to each antimicrobial were constructed with stage of production, duration of antimicrobial use and the use of 5 antimicrobial classes included as explanatory variables in each model. The greatest frequencies of resistance were observed to tetracycline, trimethoprim/sulphamethoxazole and streptomycin with the highest levels of resistance observed in isolates from first-stage weaned pigs. Differences in the types of antimicrobial drugs used were noted between long-term and short-term use farms. Categorization of farms as long- or short-term use was sufficient to predict the likely occurrence of resistance to 3 antimicrobial classes and could provide an aid in the control of resistance in the food chain. Stage of production was a significant predictor variable in all models of resistance constructed and did not solely reflect antimicrobial use at each stage. Cross-selection and co-selection for resistance was evident in the models constructed, and the use of trimethoprim/sulphonamide drugs in particular was associated with the occurrence of resistance to other antimicrobials.

A longitudinal study of Staphylococcus aureus colonization in pigs in Ireland.

The emergence of methicillin-resistant Staphylococcus aureus (MRSA) in livestock has refocused attention on S. aureus colonization and transmission in pigs. This study investigated the effect of the S. aureus colonization status of a sow on the colonization status of her piglets, and whether pigs carry the same strain of S. aureus throughout production. Nasal swabs were collected from the piglets of six healthy sows two days after birth and two days before and two days after they were moved into each production stage. The average prevalence of S. aureus colonization varied between 26% and 73%. The odds of being S. aureus positive were almost 12 times higher for piglets born to nasal-positive sows than for those born to nasal-negative sows, and three times higher again for piglets born to sows that were both nasal- and vaginal-positive. Isolates recovered from piglets immediately after birth were indistinguishable from those of the dam as determined by phenotypic and molecular typing, including microarray analysis and optical mapping. All isolates belonged to clonal complex 9 and the majority exhibited a novel spa type, t10449. The findings show that the S. aureus colonization status of the sow influences the colonization status of her piglets in the early production stages but strains carried by pigs change over time. Multiresistant S. aureus was detected, in particular post-weaning. Results suggest that sow status and management practices, including mixing of pigs and antimicrobial usage at weaning, should be considered when implementing control measures for S. aureus on a farm.

Influences on antimicrobial prescribing behaviour of veterinary practitioners in cattle practice in Ireland.

Guidelines on prudent antimicrobial use in veterinary medicine have been developed to reduce inappropriate prescribing of antimicrobials. Such guidelines focus mainly on the clinical and pharmacological indications for prescribing. A questionnaire study of veterinary surgeons engaged in cattle practice was completed to determine if non-clinical issues influence the decision to prescribe antimicrobials, and to assess if pharmacological and non-pharmacological issues influence the choice of antimicrobial prescribed. Non-clinical issues, including issues related to professional stress, influenced the prescribing decision of the majority of respondents. However, the nature of the veterinarian-client relationship did not influence the prescribing behaviour of the majority of respondents. Pharmacological and non-pharmacological issues influenced the choice of antimicrobial prescribed. The veterinary surgeon's prior experience of a drug was considered 'often' or 'always' by 95.7 per cent of respondents when making this decision. The findings of this study have implications for the recognition and management of stress within the profession, and for the development of intervention strategies to reduce inappropriate antimicrobial prescribing.

Investigation of the persistence and transmission of MRSA CC 5 in pigs following intra-nasal inoculation.

MRSA CC5 spa type t002 appears to have a broad host range, has been isolated from animals and in-contact humans in Ireland and could potentially become established in pigs in Ireland. The aims of this study were to determine if MRSA CC5 spa type t002 could persist in the tissues of the porcine upper respiratory tract following intra-nasal inoculation; to determine the relative importance of environmental and animal sources of the bacterium in the transmission cycle and to determine the importance of the pharynx as a carriage site of Staphylococcus aureus and MRSA. Twelve pigs were inoculated intra-nasally with MRSA CC5 t002. After 1 or 6 days, the inoculated pigs were removed from the contaminated environment, were washed in an antiseptic solution and placed in a clean house with a group of naive pigs (in-contact group). Another group of naive pigs was placed in the contaminated environment to assess transmission from the environment (environmental group). Nasal swabs, environmental swabs and tissue samples from the upper respiratory tract were taken for MRSA culture. Infection rates were calculated for each group of exposed pigs. MRSA persisted in the pharyngeal tissues of 6 inoculated pigs for at least 30 days and higher counts of S. aureus were found in pharyngeal tissues than in other sites. In this study we were able to demonstrate the establishment of colonisation by MRSA CC5 spa type t002 in commercially sourced pigs already colonised by S. aureus; however, colonisation was sporadic despite the inoculation of large doses. Onward transmission via pig-to-pig contact or environmental contamination was possible and a significant difference was found between the proportion of pigs infected in the environmental group and the proportion infected in the in-contact group during the first 5 days. However, no significant difference was detected in overall infection rates between the 2 groups. The tissues of the pharynx were found to carry greater numbers of S. aureus than other tissues of the upper respiratory tract; therefore, pharyngeal carriage of MRSA and S. aureus in pigs may be more significant than previously thought.

Detection of Toxoplasma gondii antigens reactive with antibodies from serum, amniotic, and allantoic fluids from experimentally infected pregnant ewes.

Toxoplasma gondii, an intracellular protozoan parasite, is one of the major causes of infectious abortion in sheep. To further understand the pathogenesis of toxoplasmosis, serum, amniotic and allantoic fluids and foetal stomach contents were collected from experimentally infected pregnant ewes to determine pathogen numbers and other markers of infection. Fifteen pregnant ewes (90 days of gestation) were each orally inoculated with 3000 sporulated oocysts of T. gondii. Serum samples were collected weekly following challenge. Amniotic and allantoic fluids and foetal stomach contents were collected at 21, 25, 28, 33 and 35 days post-infection. Characteristic placental lesions were detected in 1 of 4 challenged ewes at day 25, 3 of 4 challenged ewes at day 28 and in all challenged ewes at days 33 and 35 post-infection. T. gondii was detected only sporadically in amniotic and allantoic fluids before 35 days of infection, by real-time PCR, and only in ewes with placental lesions. At 35 days post-infection, high numbers of parasite were detected in both amniotic and allantoic fluids. An increase in the number of fluids from challenged animals with IgM and IgG was detected over time, except for IgG in allantoic fluid, which was detected in all samples from day 21 post-infection. IgG in amniotic and allantoic fluids was shown to be specific for T. gondii, and reacted with antigens with an apparent molecular mass of approximately 22 kDa and 30 kDa. Results suggest a maternal source of immunoglobulin in the allantoic fluid and a foetal source of immunoglobulin in the amniotic fluid early in infection but that both sources may contribute immunoglobulin to both fluids at a later stage.

Interferon-γ expression in trophoblast cells in pregnant ewes challenged with Chlamydophila abortus.

Pregnant ewes were challenged with Chlamydia abortus at 91-98 days of gestation and euthanised at 14, 21 and 28 days post-challenge. IFNγ mRNA labelling appeared to be co-localised with Chlamydial lipopolysaccharide within trophoblast cells in discrete areas lining the primary villi in the limbus and hilar zone of the placentomes from challenged sheep on days 21 and 28 post-infection. The presence of IFNγ was also demonstrated by immunohistochemistry. No labelling was seen in tissues from the non-infected ewes. The presence of IFNγ in trophoblast cells from infected ewes may indicate an attempt to restrict the replication of the organism and be an important trigger for the inflammatory responses that develop on the fetal side of the placenta in enzootic abortion.

Phylogenetic analysis of H and N2 genes of avian influenza viruses detected in Ireland between 2003 and 2007.

Phylogenetic analysis was performed on the haemagglutinin and neuraminidase subtype N2 genes of low-pathogenic avian influenza viruses (LPAIVs) detected in Ireland between 2003 and 2007. Nucleotide sequences were compared to previously published sequences from the National Centre for Biotechnology Information. Sequences from viruses of the same subtype isolated in different years were compared to examine the possibility that LPAIVs may have been maintained in Ireland from year to year. All viruses had closest identity with published sequences of European lineage, supporting the conclusion that LPAIVs had been introduced to Ireland by dabbling ducks that had migrated from Europe. The data suggested that different subtypes of virus had been introduced each year. However, there was evidence that some LPAIVs may have been maintained in the sedentary waterfowl population for consecutive seasons. Furthermore, almost identical H6 and H10 sequences with different N types were found in isolates from the same season, suggesting that reassortment had occurred.

Detection and quantification of leptospires in urine of dogs: a maintenance host for the zoonotic disease leptospirosis.

Leptospirosis is a global zoonotic disease. Pathogenic Leptospira species, the causative agent of leptospirosis, colonize the renal tubules of chronically infected maintenance hosts such as dogs, rats and cattle. Maintenance hosts typically remain clinically asymptomatic and shed leptospires into the environment via urine. In contrast, accidental hosts such as humans can suffer severe acute forms of the disease. Infection results from direct contact with infected urine or indirectly, through contaminated water sources. In this study, a quantitative real-time PCR specific for lipL32 was designed to detect the urinary shedding of leptospires from dogs. The sensitivity and specificity of the assay was evaluated using both a panel of pathogenic Leptospira species and clinical microbial isolates, and samples of urine collected from experimentally infected rats and non-infected controls. The lower limit of detection was approximately 3 genome equivalents per reaction. The assay was applied to canine urine samples collected from local dog sanctuaries and the University Veterinary Hospital (UVH) at University College Dublin. Of 525 canine urine samples assayed, 37 were positive, indicating a prevalence of urinary shedding of leptospires of 7.05%. These results highlight the need to provide effective canine vaccination strategies and raise public health awareness.

Detection and quantification of Toxoplasma gondii in ovine maternal and foetal tissues from experimentally infected pregnant ewes using real-time PCR.

A real-time PCR (rt-PCR) targeting the 529-bp repeat element (RE) of Toxoplasma gondii was used to detect and quantify the parasite burden in maternal and foetal tissues in 18 seronegative ewes infected with 3000 toxoplasma oocysts on day 90 of pregnancy. The infected ewes were sacrificed in groups of 4-6 at 21, 25, 33 and 35 days post-challenge. Ten sham inoculated pregnant ewes were used as controls. T. gondii was not detected in the control ewes or their foeti. The parasite was only detected in the maternal tissues in a few of the challenged ewes on a small number of occasions where it was identified in spleen and uterine lymph nodes. T. gondii was detected in the foetal spleen and liver at the early sacrifice times but only sporadically thereafter. In the case of amniotic, allantoic and foetal aqueous humor samples T. gondii was only detected on a small number of occasions. However, it was found in the majority of the foetal lung and placentome samples throughout the study period, while placentomes and foetal brains contained high levels of the parasite during the later stages. Histopathological examination of placentome and brain tissue from the foeti in the present study revealed a strong correlation between histopathological lesions and quantities of the parasite DNA detected. These results indicate that the cotyledonary component of the foetal membranes is the sample of choice for the diagnosis of T. gondii by rt-PCR, followed by foetal lung and brain.

Isolation rates of meticillin-resistant Staphylococcus aureus in dogs, cats and horses in Ireland.

A retrospective analysis and prospective surveillance study were conducted to determine isolation rates of meticillin-resistant Staphylococcus aureus (MRSA) in dogs, cats and horses in Ireland. Clinical samples that had been submitted to University College Dublin (UCD) for routine microbiological examination over a four-year period (2003 to 2006) were analysed in the retrospective analysis, which included clinical samples from 3866 animals. In the prospective surveillance study, samples from healthy animals presenting for elective surgery as well as from animals with a clinical presentation suggestive of MRSA infection were investigated. Animals attending 30 veterinary practices throughout Ireland and a similar population of animals presented to UCD were studied. The isolation rates for animals in the retrospective study were 1.1 per cent (32 of 2864) for dogs, 0.7 per cent (four of 619) for cats and 5.2 per cent (20 of 383) for horses. The overall isolation rate of MRSA was 1.4 per cent (56 of 3866). Isolation rates for healthy animals in the prospective study were 0.4 per cent (one of 286) for dogs and 1.7 per cent (four of 236) for horses; MRSA was not isolated from cats (0 of 47). Isolation rates for animals suspected of being infected with MRSA were 8.1 per cent (14 of 173) for dogs and 4.6 per cent (three of 65) for horses; MRSA was not isolated from cats (0 of 47).

Detection of three distinct genetic lineages in methicillin-resistant Staphylococcus aureus (MRSA) isolates from animals and veterinary personnel.

This study involved the phenotypic and molecular characterization of a population of methicillin-resistant Staphylococcus aureus isolates from animals and from veterinary personnel in Ireland. Isolates from 77 animals (dogs, n=44; cats, n=4; horses, n=29) and from 28 veterinary personnel were characterized using their antimicrobial resistance profiles and pulsed-field gel electrophoresis patterns. In addition, a representative number of these isolates (n=52) were further analysed using spa-typing techniques. The results obtained identified the presence of three distinct clonal complexes, CC5, CC8 and CC22, in both animal and human isolates. Two of these clonal complexes, CC8 and CC22, respectively, have been previously described in animals in Ireland but the presence of the third complex CC5 is a novel finding. The significance of this development, in relation to human and animal healthcare, is discussed.

Zoonotic transmission of Streptococcus equi subsp. zooepidemicus from a dog to a handler.

This is, to the best of our knowledge, the first case report to describe the apparent transmission of Streptococcus equi subsp. zooepidemicus from an infected dog to a handler who subsequently developed severe systemic infection. Characterization of the haemolytic streptococci isolated from both the patient and the dog, by phenotypic and molecular analysis, confirmed the canine and human isolates were identical.

High-level gentamicin-resistant and vancomycin-resistant Enterococcus faecium isolated from a wound in a dog.

This report describes the first clinical case, in Europe, of a high-level gentamicin-resistant and vancomycin-resistant Enterococcus faecium infection in a dog. The aim of this report is to alert the veterinary profession to high-level gentamicin-resistant and vancomycin-resistant Enterococcus faecium in companion animal patients. Multi-drug resistant strains of this pathogen are the cause of considerable problems in human hospitals and for community healthcare professionals worldwide. The potential for a similar impact within veterinary medicine is discussed.

Avian influenza viruses detected by surveillance of waterfowl in Ireland during 2003-2007.

Specimens for the detection of avian influenza virus (AIV) were collected from 1937 waterfowl on the Wexford Sloblands, a major wetland reserve in southeast Ireland, between January 2003 and September 2007. During the same period, 1404 waterfowl were sampled at other locations in Ireland. Specimens were tested either by virus isolation or real-time reverse transcriptase polymerase chain reaction (rtRT-PCR). A total of 32 isolates of AIV, comprising nine subtypes, was obtained from specimens from the Sloblands compared with just one isolate from elsewhere in Ireland. Samples from nine other waterfowl, five of which were from the Sloblands, tested positive for AIV by rtRT-PCR. Ecological factors are likely to have contributed to the higher detection rate of AIV at the Sloblands compared with the rest of Ireland. It was concluded that targeted surveillance at such sites is a cost-effective means of monitoring the circulation of new AIVs in waterfowl, whereas widespread opportunistic sampling is unproductive and wasteful of resources.

Meticillin-resistant Staphylococcus aureus in animals: a review.

The objectives of this paper are to review published data on the prevalence and epidemiology of meticillin-resistant Staphylococcus aureus (MRSA) infection and colonization in animals and to provide suggestions for preventing and controlling the problem in veterinary practice. MRSA first emerged as a serious pathogen in human medicine during the late 1970s and has been increasingly reported in animals during the past 10 years. The prevalence of MRSA in human infections varies markedly between geographical areas, being as high as 60% in parts of the USA, 40% in southern Europe but <1% in northern Europe. Epidemiological evidence, including phenotypic and molecular typing data, suggests that MRSA isolates from dogs and cats are indistinguishable from human healthcare isolates, whereas strains of MRSA isolated from horses and associated personnel are different. There is evidence that transfer of MRSA strains can occur between animals and humans and vice versa. Guidelines for the control of MRSA in animals have been drawn up by individual institutions based on those available for human MRSA infection. Risk factors for MRSA infection in animals are currently under investigation and such data are essential for the preparation of specific guidelines for control of MRSA in veterinary practice.