Combining mutation and horizontal gene transfer in a within-host model of antibiotic resistance.

Antibiotics are used extensively to control infections in humans and animals, usually by injection or a course of oral tablets. There are several methods by which bacteria can develop antimicrobial resistance (AMR), including mutation during DNA replication and plasmid mediated horizontal gene transfer (HGT). We present a model for the development of AMR within a single host animal. We derive criteria for a resistant mutant strain to replace the existing wild-type bacteria, and for co-existence of the wild-type and mutant. Where resistance develops through HGT via conjugation we derive criteria for the resistant strain to be excluded or co-exist with the wild-type. Our results are presented as bifurcation diagrams with thresholds determined by the relative fitness of the bacteria strains, expressed in terms of reproduction numbers. The results show that it is possible that applying and then relaxing antibiotic control may lead to the bacterial load returning to pre-control levels, but with an altered structure with regard to the variants that comprise the population. Removing antimicrobial selection pressure will not necessarily reduce AMR and, at a population level, other approaches to infection prevention and control are required, particularly when AMR is driven by both mutation and mobile genetic elements.

Source attributed case-control study of campylobacteriosis in New Zealand.

BACKGROUND: Following an initial reduction in human campylobacteriosis in New Zealand after the implementation of poultry food chain-focused interventions during 2006-2008, further decline has been relatively small. We report a year-long study of notified campylobacteriosis cases, incorporating a case control study combined with a source attribution study. The purpose was to generate up-to-date evidence on the relative contributions of different sources of campylobacteriosis in New Zealand. METHODS: The study approach included: • A case-control study of notified cases (aged six months or more) sampled in a major urban centre (Auckland, every second case) and a mixed urban/rural area (Manawatu/Whanganui, every case), between 12 March 2018 and 11 March 2019. • Source attribution of human campylobacteriosis cases sampled from these two regions over the study period by modelling of multilocus sequence typing data of Campylobacter jejuni and C. coli isolates from faecal samples of notified human cases and relevant sources (poultry, cattle, sheep). RESULTS: Most cases (84%) were infected with strains attributed to a poultry source, while 14% were attributed to a cattle source. Approximately 90% of urban campylobacteriosis cases were attributed to poultry sources, compared to almost 75% of rural cases. Poultry consumption per se was not identified as a significant risk factor. However specific risk factors related to poultry meat preparation and consumption did result in statistically significantly elevated odds ratios. CONCLUSIONS: The overall findings combining source attribution and analysis of specific risk factors indicate that poultry meat remains a dominant pathway for exposure and infection.

Microbial contamination in drinking water at public outdoor recreation facilities in New Zealand.

AIM: The aim of our study was to assess the presence and risk of waterborne pathogens in the drinking water of outdoor facilities in New Zealand and track potential sources of microbial contamination in water sources. METHODS AND RESULTS: A serial cross-sectional study with a risk-based sample collection strategy was conducted at 15 public campgrounds over two summer seasons (2011-2012 and 2012-2013). Drinking water supplied to these campgrounds was not compliant with national standards, based on Escherichia coli as an indicator organism, in more than half of the sampling occasions. Campylobacter contamination of drinking water at the campgrounds was likely to be of wild bird origin. Faecal samples from rails (pukeko and weka) were 35 times more likely to return a Campylobacter-positive result compared to passerines. Water treatment using ultraviolet (UV) irradiation or a combination of filtration and UV irradiation or chemicals was more likely to result in water that was compliant with the national standards than water from a tap without any treatment. The use of filters alone was not associated with the likelihood of compliance. CONCLUSIONS: Providing microbiologically safe drinking water at outdoor recreational facilities is imperative to avoid gastroenteritis outbreaks. This requires an in-depth understanding of potential sources of contamination in drinking water sources and the installation of adequate water treatment facilities. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study provides evidence that drinking water without treatment or filter-only treatment in public campgrounds is unlikely to comply with national standards for human consumption and extra water treatment measures such as UV irradiation or chemical treatment are needed.

Estimating the level of disease risk and biosecurity on commercial poultry farms in New Zealand.

Aims: To collect baseline data on the contact risk pathways and biosecurity practices of commercial poultry farms in New Zealand, investigate the relationship between the farm-level disease contact risks and biosecurity practices, and identify important poultry health concerns of producers. Methods: A cross-sectional survey of all registered New Zealand commercial poultry operations was conducted in 2016 collecting information on farm demographics, biosecurity practices, and contact risk pathways. Survey responses were used to generate an unweighted subjective disease risk score based on eight risk criteria and a subjective biosecurity score based on the frequency with which producers reported implementing seven biosecurity measures. Producer opinions towards poultry health issues were also determined. Results: Responses to the survey response were obtained from 120/414 (29.0%) producers, including 57/157 (36.3%) broiler, 33/169 (19.5%) layer, 24/55 (44%) breeder, and 6/32 (19%) other poultry production types. Median disease risk scores differed between production types (p < 0.001) and were lowest for breeder enterprises. The greatest risk for layer and broiler enterprises was from the potential movement of employees between sheds, and for breeder enterprises was the on- and off-farm movement of goods and services. Median biosecurity scores also differed between production types (p < 0.001), and were highest for breeder and broiler enterprises. Across all sectors there was no statistical correlation between biosecurity scores and disease risk scores. Producers showed a high level of concern over effectively managing biosecurity measures. Conclusions: The uptake of biosecurity measures in the commercial poultry farms surveyed was highly variable, with some having very low scores despite significant potential disease contact risks. This may be related to the low prevalence or absence of many important infectious poultry diseases in New Zealand leading farmers to believe there is a limited need to maintain good biosecurity as well as farmer uncertainty around the efficacy of different biosecurity measures. Further research is needed to understand barriers towards biosecurity adoption including evaluating the cost-effectiveness of biosecurity interventions.

The effect of transportation and lairage on faecal shedding and carcass contamination with Escherichia coli O157 and O26 in very young calves in New Zealand.

The effect of transportation and lairage on the faecal shedding and post-slaughter contamination of carcasses with Escherichia coli O157 and O26 in young calves (4-7-day-old) was assessed in a cohort study at a regional calf-processing plant in the North Island of New Zealand, following 60 calves as cohorts from six dairy farms to slaughter. Multiple samples from each animal at pre-slaughter (recto-anal mucosal swab) and carcass at post-slaughter (sponge swab) were collected and screened using real-time PCR and culture isolation methods for the presence of E. coli O157 and O26 (Shiga toxin-producing E. coli (STEC) and non-STEC). Genotype analysis of E. coli O157 and O26 isolates provided little evidence of faecal-oral transmission of infection between calves during transportation and lairage. Increased cross-contamination of hides and carcasses with E. coli O157 and O26 between co-transported calves was confirmed at pre-hide removal and post-evisceration stages but not at pre-boning (at the end of dressing prior to chilling), indicating that good hygiene practices and application of an approved intervention effectively controlled carcass contamination. This study was the first of its kind to assess the impact of transportation and lairage on the faecal carriage and post-harvest contamination of carcasses with E. coli O157 and O26 in very young calves.

Cost-benefit analyses of supplementary measles immunisation in the highly immunized population of New Zealand.

As endemic measles is eliminated from countries through increased immunisation, the economic benefits of enhanced immunisation programs may come into question. New Zealand has suffered from outbreaks after measles introductions from abroad and we use it as a model system to understand the benefits of catch up immunisation in highly immunised populations. We provide cost-benefit analyses for measles supplementary immunisation in New Zealand. We model outbreaks based on estimates of the basic reproduction number in the vaccinated population (Rv, the number of secondary infections in a partially immunised population), based on the number of immunologically-naïve people at district and national levels, considering both pre- and post-catch up vaccination scenarios. Our analyses suggest that measles Rv often includes or exceeds one (0.18-3.92) despite high levels of population immunity. We calculate the cost of the first 187 confirmed and probable measles cases in 2014 to be over NZ$1 million (∼US$864,200) due to earnings lost, case management and hospitalization costs. The benefit-cost ratio analyses suggest additional vaccination beyond routine childhood immunisation is economically efficient. Supplemental vaccination-related costs are required to exceed approximately US$66 to US$1877 per person, depending on different scenarios, before supplemental vaccination is economically inefficient. Thus, our analysis suggests additional immunisation beyond childhood programs to target naïve individuals is economically beneficial even when childhood immunisation rates are high.

Characterisation by multilocus sequence and porA and flaA typing of Campylobacter jejuni isolated from samples of dog faeces collected in one city in New Zealand.

AIMS: To investigate the prevalence of Campylobacter spp. and C. jejuni in dog faecal material collected from dog walkways in the city of Palmerston North, New Zealand, and to characterise the C. jejuni isolates by multilocus sequence typing (MLST) and porA and flaA antigen gene typing. METHODS: A total of 355 fresh samples of dogs faeces were collected from bins provided for the disposal of dog faeces in 10 walkways in Palmerston North, New Zealand, between August 2008-July 2009. Presumptive Campylobacter colonies, cultured on modified charcoal cefoperazone deoxycholate plates, were screened for genus Campylobacter and C. jejuni by PCR. The C. jejuni isolates were subsequently characterised by MLST and porA and flaA typing, and C. jejuni sequence types (ST) were assigned. RESULTS: Of the 355 samples collected, 72 (20 (95% CI=16-25)%) were positive for Campylobacter spp. and 22 (6 (95% CI=4-9)%) were positive for C. jejuni. Of the 22 C. jejuni isolates, 19 were fully typed by MLST. Ten isolates were assigned to the clonal complex ST-45 and three to ST-52. The allelic combinations of ST-45/flaA 21/porA 44 (n=3), ST-45/flaA 22/porA 53 (n=3) and ST-52/ flaA 57/porA 905 (n=3) were most frequent. CONCLUSIONS: The successful isolation of C. jejuni from canine faecal samples collected from faecal bins provides evidence that Campylobacter spp. may survive outside the host for at least several hours despite requiring fastidious growth conditions in culture. The results show that dogs carry C. jejuni genotypes (ST-45, ST-50, ST-52 and ST-696) that have been reported in human clinical cases. CLINICAL RELEVANCE: Although these results do not provide any evidence either for the direction of infection or for dogs being a potential risk factor for human campylobacteriosis, dog owners are advised to practice good hygiene with respect to their pets to reduce potential exposure to infection.

Global importation and population risk factors for measles in New Zealand: a case study for highly immunized populations.

As endemic measles is eliminated through immunization, countries must determine the risk factors for the importation of measles into highly immunized populations to target control measures. Despite eliminating endemic measles, New Zealand suffers from outbreaks after introductions from abroad, enabling us to use it as a model for measles introduction risk. We used a generalized linear model to analyze risk factors for 1137 measles cases from 2007 to June 2014, provide estimates of national immunity levels, and model measles importation risk. People of European ethnicity made up the majority of measles cases. Age is a positive risk factor, particularly 0-2-year-olds and 5-17-year-old Europeans, along with increased wealth. Pacific islanders were also at greater risk, but due to 0-2-year-old cases. Despite recent high measles, mumps, and rubella vaccine immunization coverage, overall population immunity against measles remains ~90% and is lower in people born between 1982 and 2005. Greatest measles importation risk is during December, and countries predicted to be sources have historical connections and highest travel rates (Australia and UK), followed by Asian countries with high travel rates and higher measles incidences. Our results suggest measles importation due to travel is seeding measles outbreaks, and immunization levels are insufficient to continue to prevent outbreaks because of heterogeneous immunity in the population, leaving particular age groups at risk.

Epidemiology of Escherichia coli serogroups O26, O103, O111 and O145 in very young ('bobby') calves in the North Island, New Zealand.

The prevalence and spatial distribution of Escherichia coli serogroups O26, O103, O111 and O145 in calves 70% similarity) using pulsed field gel electrophoresis. Mapping of the farms showed the presence of farms positive for O26, O103 and O145 in three important dairy producing regions of the North Island. Calves positive for O103 were more likely to be positive for O26 and vice versa (P = 0·04). Similarly, calves positive for O145 were more likely to be positive for O103 and vice versa (P = 0·03). This study demonstrates that non-O157 E. coli serogroups of public health and economic importance containing clinically relevant virulence factors are present in calves in the North Island of New Zealand.

Multidrug resistant Enterobacteriaceae in New Zealand: a current perspective.

In this article we review mechanisms and potential transmission pathways of multidrug resistance in Enterobacteriaceae, with an emphasis on extended-spectrum ß-lactamase (ESBL)-production. This provides background to better understand challenges presented by this important group of antimicrobial resistant bacteria, and inform measures aimed at prevention and control of antimicrobial resistance in general. Humans and animals interact at various levels; household pets cohabit with humans, and other animals interact with people through direct contact, as well as through the food chain and the environment. These interactions offer opportunity for bacteria such as ESBL-producers to be shared and transmitted between species and, in turn, increase the risk of zoonotic and reverse-zoonotic disease transmission. A key step in curtailing antimicrobial resistance is improved stewardship of antimicrobials, including surveillance of their use, better infection-control and prevention, and a better understanding of prescribing practice in both veterinary and medical professions in New Zealand. This will also require prospective observational studies to examine risk factors for antimicrobial resistance. Due to the interconnectedness of humans, animals and the environment actions to effect the changes required should be undertaken using a One Health approach.

Estimating Bacterial Pathogen Levels in New Zealand Bulk Tank Milk.

Zoonotic bacteria such as Campylobacter, Listeria, and Shiga toxin-producing Escherichia coli have been found in bulk tank milk in many countries, and the consumption of raw milk has been implicated in outbreaks of disease in New Zealand. Fecal contamination at milking is probably the most common source of pathogenic bacteria in bulk tank milk. Raw milk was collected from 80 New Zealand dairy farms during 2011 and 2012 and tested periodically for Campylobacter, E. coli O157, Listeria monocytogenes, and Salmonella. Milk quality data such as coliform counts, total bacterial counts, and somatic cell counts also were collected. By treating the total bacterial count as a proxy for fecal contamination of milk and utilizing farm and animal level prevalence and shedding rates of each pathogen, a predictive model for the level of pathogenic bacteria in bulk tank raw milk was developed. The model utilizes a mixture distribution to combine the low level of contamination inherent in the milking process with isolated contamination events associated with significantly higher pathogen levels. By simulating the sampling and testing process, the predictive model was validated against the observed prevalence of each pathogen in the survey. The predicted prevalence was similar to the observed prevalence for E. coli O157 and Salmonella, although the predicted prevalence was higher than that observed in samples tested for Campylobacter.

Nationwide prevalence and risk factors for faecal carriage of Escherichia coli O157 and O26 in very young calves and adult cattle at slaughter in New Zealand.

Nationwide prevalence and risk factors for faecal carriage of Escherichia coli O157 and O26 in cattle were assessed in a 2-year cross-sectional study at four large slaughter plants in New Zealand. Recto-anal mucosal swab samples from a total of 695 young (aged 4-7 days) calves and 895 adult cattle were collected post-slaughter and screened with real-time polymerase chain reaction (PCR) for the presence of E. coli O157 and O26 [Shiga toxin-producing E. coli (STEC) and non-STEC]. Co-infection with either serogroup of E. coli (O157 or O26) was identified as a risk factor in both calves and adult cattle for being tested real-time PCR-positive for E. coli O157 or O26. As confirmed by culture isolation and molecular analysis, the overall prevalence of STEC (STEC O157 and STEC O26 combined) was significantly higher in calves [6·0% (42/695), 95% confidence interval (CI) 4·4-8·1] than in adult cattle [1·8% (16/895), 95% CI 1·1-3·0] (P < 0·001). This study is the first of its kind in New Zealand to assess the relative importance of cattle as a reservoir of STEC O157 and O26 at a national level. Epidemiological data collected will be used in the development of a risk management strategy for STEC in New Zealand.

Network analysis of wildlife translocations in New Zealand.

AIMS: To identify network measures with relevance to disease spread in a network of movements derived from the Department of Conservation (DOC) translocation records from 1970 to mid-2014, and to identify conservation sites that should be prioritised for surveillance activities and improvements to data collection to make the best use of network analysis techniques in the future. METHODS: Data included the source and destination of translocated specimens, the species and the dates the translocations were expected to occur. The data were used to construct a directed, non-weighted network in which a translocation event represented a tie in the network. Network density, in-degree (movements entering a node of interest) and out-degree (movements leaving a node of interest) and reciprocity were calculated. RESULTS: The data analysed consisted of 692 unique translocations between 307 sites, with the majority (518; 73%) being for birds. The constructed network for bird, reptile and frog translocations comprised 260 nodes, with 34/260 (13%) having two-way movements and 47/260 (18%) non-reciprocal movements. The median degree score (sum of in- and out-degree) was two (min 0, max 36) with a mean of 3.5 in a right skewed distribution. Most sites acted as receivers or senders of consignments with only a few having both high in- and high out-degree, and thus had characteristics that made them sites of interest for surveillance activities. These included the National Wildlife Centre at Mount Bruce, Tiritiri Matangi Island and Te Kakahu (Chalky Island). CONCLUSIONS: The presence of linking sites that join larger clusters within the network creates the potential for rapid disease spread if a pathogen were to be introduced. The important sites that supply or receive specimens for translocations are already well recognised by those performing translocations in New Zealand, and this paper provides further information by quantifying their role within the network.

Diversity and relatedness of Shiga toxin-producing Escherichia coli and Campylobacter jejuni between farms in a dairy catchment.

The aim of this study was to examine the population structure, transmission and spatial relationship between genotypes of Shiga toxin-producing Escherichia coli (STEC) and Campylobacter jejuni, on 20 dairy farms in a defined catchment. Pooled faecal samples (n = 72) obtained from 288 calves were analysed by real-time polymerase chain reaction (rtPCR) for E. coli serotypes O26, O103, O111, O145 and O157. The number of samples positive for E. coli O26 (30/72) was high compared to E. coli O103 (7/72), O145 (3/72), O157 (2/72) and O111 (0/72). Eighteen E. coli O26 and 53 C. jejuni isolates were recovered from samples by bacterial culture. E. coli O26 and C. jejuni isolates were genotyped using pulsed-field gel electrophoresis and multilocus sequence typing, respectively. All E. coli O26 isolates could be divided into four clusters and the results indicated that E. coli O26 isolates recovered from calves on the same farm were more similar than isolates recovered from different farms in the catchment. There were 11 different sequence types of C. jejuni isolated from the cattle and 22 from water. An analysis of the population structure of C. jejuni isolated from cattle provided evidence of clustering of genotypes within farms, and among groups of farms separated by road boundaries.

Seasonality of Campylobacter jejuni isolates associated with human campylobacteriosis in the Manawatu region, New Zealand.

A 9-year time-series of genotyped human campylobacteriosis cases from the Manawatu region of New Zealand was used to investigate strain-type seasonality. The data were collected from 2005 to 2013 and the samples were multi-locus sequence-typed (MLST). The four most prevalent clonal complexes (CCs), consisting of 1215 isolates, were CC48, CC21, CC45 and CC61. Seasonal decomposition and Poisson regression with autocorrelated errors, were used to display and test for seasonality of the most prevalent CCs. Of the four examined CCs, only CC45 showed a marked seasonal (summer) peak. The association of CC45 with summer peaks has been observed in other temperate countries, but has previously not been identified in New Zealand. This is the first in-depth study over a long time period employing MLST data to examine strain-type-associated seasonal patterns of C. jejuni infection in New Zealand.

Evaluation of the representativeness of a sentinel surveillance site for campylobacteriosis.

It is important to assess the suitability of sentinel sites for human disease; however, there have been few publications documenting the process of formal evaluation. We describe an approach to examining the representativeness of a single sentinel site employed for campylobacteriosis surveillance and source attribution, utilizing a selection of data sources and statistical comparisons of demographic, epidemiological and pathogen genotyping data across selected regions of New Zealand. Our findings showed that while this region captured the national variability in many variables, for example by containing sizable urban and rural populations, the relative frequency of these features did vary from other regions of New Zealand. We discuss the value of choosing a sentinel site that represents the national distribution of key variables, compared to a site that captures the broad features of the wider population, but provides greater power for the monitoring of sub-populations.

Quantification of historical livestock importation into New Zealand 1860-1979.

AIMS: To quantify the numbers of live cattle, sheep and poultry imported into New Zealand and, where possible, their country of origin from 1860 to 1979. METHODS: Information on the origin and number of live animal importations into New Zealand was collected for cattle, sheep and poultry for the period 1868-1979 from the annual reports compiled by the New Zealand Registrar General's Office, Government Statistician's Office, Census and Statistics Office, Census and Statistics Department, Customs Department and Department of Statistics. Census data from 1851 to 1871 were also used to estimate the livestock population during this period. The number of animals imported and the mean population for each species in a decade were determined, and the major countries of origin were identified. RESULTS: A large number of cattle (53,384) and sheep (604,525) were imported in the 1860s, and then there was a marked reduction in importations. Live poultry were imported in relatively small numbers (20,701) from 1880 to 1939, then 1,564,330 live poultry were imported between 1960 and 1979. Australia was the predominant country of origin for sheep between 1868 and 1959 (51,347/60,918; 84.3%) and of cattle between 1868 and 1979 (10,080/15,157; 66.5%). Only 6,712 (11.0%) sheep and 3,909 (25.8%) cattle were imported from the United Kingdom over the same periods, and even fewer from other countries. CONCLUSIONS: The collated data and historical reports show that from 1860 to 1979 Australia has been the main source of livestock introduced into New Zealand. The pattern of importation showed that large numbers of cattle and sheep were initially imported in the 1860s, probably in response to rapid agricultural expansion. Thereafter importations continued at much reduced numbers. In contrast, relatively small numbers of poultry were introduced until the 1960s when large numbers were imported as part of the development of a modern high-production industry. The overall pattern for both cattle and sheep was of a bottleneck event, as initially a relatively limited number of animals arrived from outside populations, followed by population expansion with ongoing but limited immigration (admixture). Investigation into the genetic population structure of New Zealand's cattle and sheep, as well as their host-associated microorganisms, could reflect the impact of these early historical events.

Distribution of Escherichia coli strains harbouring Shiga toxin-producing E. coli (STEC)-associated virulence factors (stx1, stx2, eae, ehxA) from very young calves in the North Island of New Zealand.

The objective of this study was to determine the distribution of Shiga toxin-producing Escherichia coli (STEC) virulence markers (stx1, stx2, eae, ehxA) in E. coli strains isolated from young calves aged fewer than 7 days (bobby calves). In total, 299 recto-anal mucosal swabs were collected from animals at two slaughter plants and inoculated onto tryptone bile X-glucuronide and sorbitol MacConkey agar supplemented with cefixime and potassium tellurite. Isolates were analysed using multiplex polymerase chain reaction to detect stx1, stx2, eae and ehxA genes. The most common combination of virulence markers were eae, ehxA (n = 35) followed by eae (n = 9). In total, STEC and atypical enteropathogenic E. coli (aEPEC) were isolated from 8/299 (2·6%) and 37/299 (12·3%) calves, respectively. All the isolates could be assigned to 15 genotype clusters with >70% similarity cut-off using XbaI pulsed-field gel electrophoresis. It may be concluded that healthy calves from the dairy industry are asymptomatic carriers of a diverse population of STEC and aEPEC in New Zealand.

Campylobacteriosis in returning travellers and potential secondary transmission of exotic strains.

SUMMARY: Multilocus sequence types (STs) were determined for 232 and 737 Campylobacter jejuni/coli isolates from Dutch travellers and domestically acquired cases, respectively. Putative risk factors for travel-related campylobacteriosis, and for domestically acquired campylobacteriosis caused by exotic STs (putatively carried by returning travellers), were investigated. Travelling to Asia, Africa, Latin America and the Caribbean, and Southern Europe significantly increased the risk of acquiring campylobacteriosis compared to travelling within Western Europe. Besides eating chicken, using antacids, and having chronic enteropathies, we identified eating vegetable salad outside Europe, drinking bottled water in high-risk destinations, and handling/eating undercooked pork as possible risk factors for travel-related campylobacteriosis. Factors associated with domestically acquired campylobacteriosis caused by exotic STs involved predominantly person-to-person contacts around popular holiday periods. We concluded that putative determinants of travel-related campylobacteriosis differ from those of domestically acquired infections and that returning travellers may carry several exotic strains that might subsequently spread to domestic populations even through limited person-to-person transmission.

Temporal and farm-management-associated variation in faecal pat prevalence of Arcobacter spp. in ruminants.

In a 2-year longitudinal study of adult animals on 15 dairy farms and four sheep farms in Lancashire, UK, Arcobacter spp. were isolated from all farms although not at every sampling occasion. Faecal samples were collected and cultured using standard techniques for isolation of campylobacters. Assignment to species was via PCR assays. Apparent prevalence of Arcobacter spp. was higher in dairy cattle compared to sheep (40.1% vs. 8%, P < 0.001) and in housed cattle compared to cattle at pasture (50.1% vs. 20.9%, P < 0.001). This was reflected in the higher prevalence observed in herds that were housed (n = 4) all year compared to herds that grazed cattle on pasture in the summer and housed cattle in the winter (n = 11) (55.5% vs. 36%, P < 0.001). In the case of sheep, peak prevalence was observed in autumn with increased prevalence also being associated with improving pasture quality. There was an apparent inverse association between the faecal pat prevalence of Arcobacter spp. and Campylobacter jejuni although this may in part be an artefact of laboratory test method sensitivity, whereby a relative increase in the frequency of one bacterial species would reduce the sensitivity of detecting the other.