Escherichia coli isolates from extraintestinal organs of livestock animals harbour diverse virulence genes and belong to multiple genetic lineages.

Escherichia coli, the most common cause of bacteraemia in humans in the UK, can also cause serious diseases in animals. However the population structure, virulence and antimicrobial resistance genes of those from extraintestinal organs of livestock animals are poorly characterised. The aims of this study were to investigate the diversity of these isolates from livestock animals and to understand if there was any correlation between the virulence and antimicrobial resistance genes and the genetic backbone of the bacteria and if these isolates were similar to those isolated from humans. Here 39 E. coli isolates from liver (n=31), spleen (n=5) and blood (n=3) of cattle (n=34), sheep (n=3), chicken (n=1) and pig (n=1) were assigned to 19 serogroups with O8 being the most common (n=7), followed by O101, O20 (both n=3) and O153 (n=2). They belong to 29 multi-locus sequence types, 20 clonal complexes with ST23 (n=7), ST10 (n=6), ST117 and ST155 (both n=3) being most common and were distributed among phylogenetic group A (n=16), B1 (n=12), B2 (n=2) and D (n=9). The pattern of a subset of putative virulence genes was different in almost all isolates. No correlation between serogroups, animal hosts, MLST types, virulence and antimicrobial resistance genes was identified. The distributions of clonal complexes and virulence genes were similar to other extraintestinal or commensal E. coli from humans and other animals, suggesting a zoonotic potential. The diverse and various combinations of virulence genes implied that the infections were caused by different mechanisms and infection control will be challenging.

Q fever diagnosis in domestic ruminants: comparison between complement fixation and commercial enzyme-linked immunosorbent assays.

Q fever is an important zoonotic disease caused by infection with the bacterium Coxiella burnetii. Veterinary diagnostic laboratories, including the Veterinary Laboratories Agency (VLA) in England and Wales, have traditionally relied on the complement fixation test (CFT) for serological diagnosis. However, Q fever has assumed greater significance in recent years following several large human outbreaks linked to exposure to infected ruminants and it is essential that more reliable tests are introduced to detect the presence of C. burnetii infection in animals. The objective of the current study was to evaluate the performance of 3 commercially available enzyme-linked immunosorbent assays (ELISAs) for detection of antibodies to C. burnetii and to compare the findings with the CFT using a sample panel of 548 sera from sheep, goats, and cattle, including animals of known disease status. The statistical analysis using TAGS (test accuracy in the absence of a gold standard) software and receiver operating characteristic techniques demonstrated that the 3 ELISAs all showed improved sensitivity over the CFT. The test based on ovine antigen demonstrated the best overall performance and therefore, the VLA has adopted this test for routine use.

Temporal patterns and risk factors for Escherichia coli O157 and Campylobacter spp, in young cattle.

Escherichia coli O157 and Campylobacter jejuni and Campylobacter coli are zoonotic pathogens originating from farm animals. Cattle are the main reservoir for E. coli O157 and also contribute to human cases of campylobacteriosis through contaminated milk, direct contact, and environmental contamination. Thirty groups of young cattle on 30 farms were observed for 7 months and sampled on 4 to 6 separate occasions for E. coli O157 and C. jejuni/coli to characterize shedding patterns and identify risk factors. The within herd prevalence of E. coli O157 per sampling occasion ranged from 0 to 60% (mean = 24%) and average Campylobacter spp. within herd prevalence was 47% ranging from 0 to 100%. The prevalence of E. coli O157-positive herds declined with a linear trend throughout the study from 100 to 38% (OR: 0.5, P < 0.01), whereas time in the study was not significantly associated with Campylobacter prevalence (P = 0.13). Larger herds were more likely to be positive with either or both agents, whereas the number of suckler calves on the farm reduced the risk of both organisms (OR: 0.4/0.6, P < 0.01). Poultry on the premises reduced the risk of E. coli O157, but was not associated with Campylobacter. Emptying and cleaning the water troughs more often than once monthly reduced the risk of detecting Campylobacter and cattle sourced by private water supplies were more likely to be Campylobacter positive. No drinking water management practices were associated with E. coli O157. The risk of detecting both organisms were almost five times higher when the cattle were housed indoors (OR: 4.9, P = 0.03).

Risk factors for Campylobacter jejuni and Campylobacter coli in young cattle on English and Welsh farms.

Campylobacter jejuni and C. coli are the most prevalent causes of bacterial diarrhoea in most of the Western World. In Great Britain, the source remains unknown for the majority of cases, though poultry is considered the main source of infection. Molecular typing methods identify cattle as a potential source of a proportion of the non-source-attributed cases, mainly through direct contact, environmental contamination or milk, but little is known about the epidemiology of Campylobacter in cattle. A cross-sectional study was undertaken on young cattle 3-17 months of age on 56 cattle farms in England and Wales to identify association between the presence of C. jejuni and C. coli and farm characteristics and management practices. Campylobacter was detected on 62.5% of the farms and the presence of dairy cows (OR: 3.7, CI(95%): 1.2; 11.7), indoor housing (OR: 4.6, CI(95%): 1.8; 12.0), private water supply (OR: 2.5, CI(95%): 1.2; 5.4), presence of horses (OR: 3.2, CI(95%):1.5; 6.9) and feeding hay (OR: 2.9, CI(95%):1.6; 5.5) were associated with detection. The model's goodness-of-fit was improved when herd size was forced in the model without being statistically significant (p=0.34).

Investigation into the effectiveness of pooled fecal samples for detection of verocytotoxin-producing Escherichia coli O157 in cattle.

Verocytotoxin-producing Escherichia coli O157 (VTEC O157) may cause severe illness in people. Cattle are regarded as an important source of VTEC O157, and in an outbreak investigation, there is a necessity to establish whether or not the putative contact herd shares infection with the human case. The effectiveness of a herd investigation is impacted by the number of samples required, which will influence the time taken to collect samples and then process these in the laboratory. The objective of this study was to investigate the effectiveness of pooled sampling for detecting VTEC O157 in cattle herds in the United Kingdom. On farm 1, 150 individual fecal samples were collected during the course of a VTEC O157 outbreak investigation. One-gram and 10-g subsamples were tested from each individual sample. Once the culture results of the individual sample were known, pools comprising 5 and 10 individual samples were formed, with each pool containing a known number of positive samples. This data showed that the sensitivity of pooled sampling depended upon the proportion of positive samples in the pool. Further samples were collected from 2 more infected farms (2 and 3). Each individual sample was tested in duplicate. Pools of 5 feces were formed on-farm, and half the number of pooled feces were tested as individual feces. There was no significant difference between the number of cultures required for pooled sampling, as was the same for individual sampling, and therefore pooling did not improve the effectiveness of detection of VTEC O157.

Identification of management risk factors for VTEC O157 in young-stock in England and Wales.

We conducted a cross-sectional study on 255 cattle farms in England and Wales to identify risk factors for verocytotoxin-producing E. coli O157 (VTEC). Exposure variables were collected at the levels of the farm and of the group of young-stock within the farms. On each farm a group of young-stock (6-18 months of age) was sampled to establish VTEC status. In our multiple logistic regression, farm VTEC status was associated with access to springs (OR: 0.31, CI95%: 0.12, 0.78) and assessing the wetness of the bedding material less frequently than daily (OR: 3.89 CI95%: 1.5, 10.2). At group-level we found no associated risk factors for animals housed outdoors in fields. Significant for groups housed in pens were wet bedding (wet OR: 3.43, CI95%: 1.3, 9.4; very wet OR: 4.24, CI95%: 1.2, 14.6), number of animals in the group (10-15 OR: 2.72, CI95%: 0.75, 9.9, 16-24, OR: 3.78, CI95%: 1.2, 12.3; >25 OR: 3.78, CI95%: 1.1, 12.7) and feeding straw (OR: 2.29, CI95%: 1.2, 5.5).