Evaluation of efficacy of a biofilm-embedded bacteria-based vaccine against staphylococcal mastitis in sheep-A randomized, placebo-controlled field study.

Our objective was to evaluate the efficacy of a vaccine against staphylococcal mastitis in 5 dairy sheep farms, with 316 ewes in the vaccinated (V) group and 307 in the control (C) group studied throughout a lactation period. Two administrations of the vaccine were performed during the last stage of gestation of ewes. Starting 15 d after lambing and at monthly intervals thereafter, up to 9 milk samplings were performed for bacteriological and cytological examinations. Staphylococcal isolates recovered were examined for biofilm formation. Blood samples were collected for measurement of IgG poly-N-acetylglucosamine-specific antibodies. The most frequently isolated bacteria were staphylococci: 56.4 and 76.1%, respectively, of total isolates recovered from ewes of group V and C, respectively; staphylococci as causal agents of mastitis were isolated less frequently from V (5.3%) than in ewes in C (10.3%). Among mastitis-associated staphylococcal isolates recovered from V ewes, a smaller proportion was biofilm-forming than among ones from C: 53.2% versus 74.9% of isolates; biofilm-forming staphylococci as causal agents of mastitis were isolated less frequently from ewes in group V (2.3%) than in ewes in group C (6.0%). Anti-poly-N-acetylglucosamine-specific antibody values increased in V ewes and were higher than in C; a greater proportion of ewes with low antibody titers developed staphylococcal mastitis (41.4%) than of V ewes with high antibody titers (17.0%). Incidence risk of mastitis, staphylococcal mastitis, and biofilm-associated staphylococcal mastitis was smaller in V than in C: 36.7, 17.1, and 8.0% versus 44.3, 30.9, and 18.9%, respectively. The first case of staphylococcal mastitis occurred later in V than in C: third versus second sampling point. Overall, efficacy of the vaccine was 44.6% for staphylococcal mastitis, 57.7% for biofilm-associated staphylococcal mastitis, 33.1% for staphylococcal intramammary infection, and 51.5% for biofilm-associated staphylococcal intramammary infection. Nevertheless, vaccination should not be the only means for controlling mastitis; other udder health management measures should be included therein to improve control of the infection.

Extensive countrywide field investigation of subclinical mastitis in sheep in Greece.

The objectives of this work were (1) to investigate prevalence of subclinical mastitis, (2) to identify etiological agents involved, and (3) to study factors potentially predisposing ewes to subclinical mastitis. Milk samples were collected from 2,198 ewes in 111 farms with a total population of 35,925 ewes, in all 13 administrative regions of Greece, for bacteriological and cytological examination. Prevalence of subclinical mastitis was 0.260. Main etiological agents were staphylococci (Staphylococcus aureus and coagulase-negative species), which accounted for 0.699 of all isolates recovered; prevalence of staphylococcal mastitis was 0.191. In a multivariable mixed-effects analysis, the primary factor found to be associated with increased prevalence of subclinical mastitis was the management system practiced in flocks (flocks under a semi-intensive system had the highest prevalence). Other factors that were included in the multivariable model were the stage of lactation period (ewes in the 2nd month postpartum showed the highest prevalence) and application of postmilking teat dipping. In contrast, measures taken at the end of a lactation period (e.g., intramammary administration of antimicrobial agents) were not found to have an effect on prevalence of subclinical mastitis. The results confirmed the significance of subclinical mastitis as a frequent problem of ewes, with staphylococci as the primary etiological agent. The findings confirm the multifactorial nature of subclinical mastitis and indicate that its control should rely on many approaches.

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.

Molecular epidemiology and phylogeny reveal complex spatial dynamics in areas where canine parvovirus is endemic.

Canine parvovirus type 2 (CPV-2) is a severe enteric pathogen of dogs, causing high mortality in unvaccinated dogs. After emerging, CPV-2 spread rapidly worldwide. However, there is now some evidence to suggest that international transmission appears to be more restricted. In order to investigate the transmission and evolution of CPV-2 both nationally and in relation to the global situation, we have used a long-range PCR to amplify and sequence the full VP2 gene of 150 canine parvoviruses obtained from a large cross-sectional sample of dogs presenting with severe diarrhea to veterinarians in the United Kingdom, over a 2-year period. Among these 150 strains, 50 different DNA sequence types (S) were identified, and apart from one case, all appeared unique to the United Kingdom. Phylogenetic analysis provided clear evidence for spatial clustering at the international level and for the first time also at the national level, with the geographical range of some sequence types appearing to be highly restricted within the United Kingdom. Evolution of the VP2 gene in this data set was associated with a lack of positive selection. In addition, the majority of predicted amino acid sequences were identical to those found elsewhere in the world, suggesting that CPV VP2 has evolved a highly fit conformation. Based on typing systems using key amino acid mutations, 43% of viruses were CPV-2a, and 57% CPV-2b, with no type 2 or 2c found. However, phylogenetic analysis suggested complex antigenic evolution of this virus, with both type 2a and 2b viruses appearing polyphyletic. As such, typing based on specific amino acid mutations may not reflect the true epidemiology of this virus. The geographical restriction that we observed both within the United Kingdom and between the United Kingdom and other countries, together with the lack of CPV-2c in this population, strongly suggests the spread of CPV within its population may be heterogeneously subject to limiting factors. This cross-sectional study of national and global CPV phylogeographic segregation reveals a substantially more complex epidemic structure than previously described.

Echocardiographic reference intervals in healthy UK deerhounds and prevalence of preclinical dilated cardiomyopathy: a prospective, longitudinal study.

BACKGROUND: Sighthounds have high echocardiographic (ECHO) left ventricular volumes. Establishing robust breed-specific ECHO reference intervals (RI) for screening is important. End-diastolic volume index (EDVI), end-systolic volume index (ESVI) and ejection fraction (EF) reference ranges derived by Simpson's method of discs are not available for deerhounds. The influence of sex or body weight (BW) on left ventricular diameter during diastole (LVDd) and systole (LVDs) has never been reported. OBJECTIVES: Prospectively determine ECHO RI and assess prevalence of dilated cardiomyopathy (DCM) in healthy UK deerhounds. ANIMALS: Ninety-nine deerhounds. METHODS: Deerhounds scored on ECHO and ECG variables then classified as normal (NORM), equivocal (EQUIV) or affected (AFF) with DCM. Fifty-nine NORM deerhounds used to determine ECHO RI. RESULTS: Prevalence of DCM was 21.6%. There were significant differences in BW (P<0.001), LVDd (P<0.001) and LVDs (P<0.05) between female and male deerhounds. Cut-off values for EDVI (≥140.2 mL/m2: 79% sensitivity/97% specificity), ESVI (≥71.9 mL/m2: 94.7% sensitivity/94.2% specificity) and EF (≤42.1%: 84.2% sensitivity/92.8% specificity) were proposed to help diagnose DCM. The most reliable ECHO variables to identify AFF dogs were LVDs indexed to BW by allometric scaling and ESVI; one of the least reliable was sphericity index. Ventricular arrhythmias (VA) were identified in 13.6% of the population, with the highest prevalence in AFF deerhounds (42%). CONCLUSIONS: Preclinical DCM in deerhounds is common and VA may be associated with DCM. Healthy deerhounds have higher LVDd, LVDs and EDVI compared with other breeds. This study provides ECHO RIs for deerhounds; sex or BW RIs should be used when screening.

Molecular epidemiology and genetic diversity of Campylobacter jejuni in ruminants.

Multi-locus sequence typing was performed on 1003 Campylobacter jejuni isolates collected in a 2-year longitudinal study of 15 dairy farms and four sheep farms in Lancashire, UK. There was considerable farm-level variation in occurrence and prevalence of clonal complexes (CC). Clonal complexes ST61, ST21, ST403 and ST45 were most prevalent in cattle while in sheep CC ST42, ST21, ST48 and ST52 were most prevalent. CC ST45, a complex previously shown to be more common in summer months in human cases, was more prevalent in summer in our ruminant samples. Gene flow analysis demonstrated a high level of genetic heterogeneity at the within-farm level. Sequence-type diversity was greater in cattle compared to sheep, in cattle at pasture vs. housed, and in isolates from farms on the Pennines compared to the Southern Fylde. Sequence-type diversity was greatest in isolates belonging to CC ST21, ST45 and ST206.

Temporal and farm-management-associated variation in the faecal-pat prevalence of Campylobacter jejuni in ruminants.

In a 2-year longitudinal study of adult animals on 15 dairy farms and four sheep farms in Lancashire, UK. C. jejuni was isolated from all farms, although not on every occasion. Faecal samples were collected and cultured using standard techniques for isolation of Campylobacter. Assignment to species was via PCR assays. Peak prevalence of C. jejuni in both cattle and sheep was observed during the summer and in cattle this apparent seasonality was associated with grazing pasture [odds ratio (OR) 2.14], while in sheep it was independent of grazing. Increased prevalence was associated with increased milk yield (OR 1.05) and herd size (OR 1.01) in dairy cattle, and with increased stocking density (OR 1.29) and pasture quality (OR 2.16) in sheep. There was considerable variation in prevalence between farms but no evidence of large-scale spatial variation. The association between C. jejuni prevalence and diet in dairy cattle deserves further investigation.

Effect of dietary restriction on body condition, composition and welfare of overweight and obese pony mares.

REASONS FOR PERFORMING STUDY: Increased prevalence of obesity among UK horses and ponies demands evidence-based advice to promote weight loss. HYPOTHESIS: Restriction of dry matter intake (DMI) to 1% of body mass (BM, 67% [corrected] of predicted maintenance digestible energy [DE] requirements) would promote weight loss without compromise to health. METHODS: Five mature (mean ± s.e. 10 ± 2 years), overweight/obese pony mares (BM, 257 ± 20 kg: body condition score [BCS] 6.8/9 ± 0.5) were studied over 12 weeks. Animals were individually housed. Daily provision of a chaff-based, complete diet (measured DE, 8.5 MJ/kg DM) was restricted to 1% of actual BM as DMI daily. BCS, girth measurements and ultrasound-derived measures of subcutaneous fat depth overlying the gluteal region and 12th intercostal space (rib-eye) were recorded weekly. Body fat content was estimated at the beginning and end of the study by deuterium oxide dilution methods. Clinical biochemistry was monitored weekly. Behaviour was observed (24 h, 3/5 ponies) on 3 occasions. RESULTS: BM decreased by 4.3 ± 1.1% during the first week and thereafter by 0.7 ± 0.1% of BM at end of Week 1 each week. BCS remained constant. Heart and belly girths, rump width and subcutaneous fat depth at rib-eye decreased significantly with time and BM. Fat comprised 45 ± 19% of BM loss. Fatter animals lost relatively more fat. With decreased feeding activity, time spent in 'play' and rest increased by 36 ± 11% and 438 ± 95%, respectively. CONCLUSIONS: This plane of nutrition resulted in an overall rate of weight loss of 1% of outset BM weekly. BCS was not a useful index of early weight loss but heart and belly girths and subcutaneous rib-eye fat were identified as alternative markers. POTENTIAL RELEVANCE: This study provides an evidence-base for the management of weight loss in obese animals, especially those for which exercise may be contra-indicated.

Balloon valvuloplasty for treatment of pulmonic stenosis in English Bulldogs with an aberrant coronary artery.

BACKGROUND: English Bulldogs (EB) with pulmonic stenosis (PS) sometimes have an aberrant coronary artery (CA) type R2A encircling the pulmonary artery (PA). Balloon valvuloplasty (BV) is treatment of choice for severe PS, but is considered to be contraindicated in dogs with aberrant CA. HYPOTHESIS: Conservative BV in EB with aberrant CA is safe and improves clinical signs and quality of life. ANIMALS: Four client-owned EB with severe PS were retrospectively reviewed/analysed. METHODS: Retrospective study: Case records, echocardiography, BV, and follow-up investigations of EB diagnosed with severe PS and treated with BV were reviewed. The ratios of PA to aortic (Ao) velocity time integral (VTI) were calculated to assess progression/improvement of PS. RESULTS: An aberrant CA was confirmed on angiography in all EB. Conservative BV was performed, using a balloon of the size of the PA annulus or smaller (0.6-1 x PA annulus size). All dogs survived the procedure, but only a mild reduction in pressure gradient was achieved. There was an improvement in PA to Ao VTI in 3 of 4 dogs (P < or = .017), which were free of evidence of congestive heart failure (CHF) 5, 10, and 15 months after BV. One dog that had right-sided CHF when BV was performed died due to progressive right-sided CHF within 3 months. CONCLUSIONS: Conservative BV in EB might be safe and might improve quality and quantity of life.

Restoration of arterial oxygen tension in horses recovering from general anaesthesia.

BACKGROUND: Arterial hypoxaemia is common in anaesthetised horses, but little information exists regarding restoration of arterial oxygen tension (PaO2 ) during recovery from anaesthesia, or if intra-operative management factors exert any longer-term effect. OBJECTIVES: To evaluate PaO2 in horses recovering from general anaesthesia up to 1 h after resuming standing. STUDY DESIGN: Prospective observational clinical cohort study. METHODS: Systemically healthy adult horses undergoing inhalational general anaesthesia for elective surgical procedures were studied. Arterial blood samples were obtained anaerobically prior to pre-anaesthetic medication, at end of anaesthesia, immediately following positioning in the recovery box, then at 10-min intervals until standing. Additionally, samples were taken when horses achieved sternal recumbency, at standing (STAND) and 1 h after standing (STAND+1). Data were analysed using ANOVA and mixed-effects linear regression, with significance set at P<0.05. RESULTS: Data from one hundred and two horses were analysed. Forty horses received controlled mandatory ventilation (CMV) throughout anaesthesia, 47 breathed spontaneously (SV) and 15 initially breathed spontaneously before CMV was imposed (S-CMV). Overall, PaO2 , P(A-a)O2 and PaCO2 remained significantly lower than baseline at STAND+1 (P<0.01). CMV resulted in higher PaO2 at the end of anaesthesia (P = 0.03) and during early recovery (P<0.01) than SV. Only in group S-CMV did PaO2 , P(A-a)O2 and PaCO2 return to baseline values at STAND+1. Highest PaO2 values associated with CMV were also associated with early recovery apnoea. MAIN LIMITATIONS: Non-standardised anaesthetic management, temporal and quantitative variation in oxygen delivery during early recovery and lack of control group where oxygen was electively withheld during recovery. CONCLUSIONS: Controlled mandatory ventilation results in better pulmonary function in horses as assessed by PaO2 , P(A-a)O2 and PaCO2 , an effect enhanced by an initial period of SV and still evident 1 h after standing. High PaO2 values may contribute to early recovery apnoea but this does not adversely affect outcome. The Summary is available in Portuguese - see Supporting Information.

Study of potential environmental factors predisposing ewes to subclinical mastitis in Greece.

Objectives of the work presented herewith were to investigate association of prevalence of subclinical mastitis with environmental (climatic and topographic) factors and to identify factors potentially predisposing ewes to the disease. Milk samples were collected from 2198 sheep in 111 farms, in all 13 administrative regions of Greece, for bacteriological and cytological examination. Data on farm location were collected in the field using hand-held Global Positioning System Garmin units. The geo-references were resolved to specific farm level. Prevalence of subclinical mastitis was 0.260. Main aetiological agents were staphylococci (Staphylococcus aureus and coagulase-negative species), which accounted for 0.699 of all isolates recovered. In a multivariable mixed-effects analysis, the two environmental variables found to be associated with increased prevalence of subclinical mastitis were the minimum temperature of coldest month (coefficient: -0.084 ± 0.033, P = 0.014) and the mean temperature for 30 days prior to sampling date (coefficient: 0.031±0.014, P = 0.029).

Use of geographical information system and ecological niche modelling for predicting potential space distribution of subclinical mastitis in ewes.

Objective was to recognise areas potentially of high risk for increased frequency of subclinical mastitis in ewes. Milk samples were collected, for bacteriological and cytological examination, from 2198 clinically healthy ewes in 111 farms in all administrative regions of Greece. Data on farms were located in the field using hand-held Global Positioning System Garmin units. Collected data were analysed by an Ecological Niche Model under the framework of a geographic information system. Two separate analyses were performed: one for subclinical mastitis independently of causal agent (prevalence in population sampled: 0.260) and one for subclinical mastitis caused specifically by slime-producing staphylococci (prevalence in population sampled: 0.153). A model was constructed in which sheep farms were divided into two clusters, according to prevalence of subclinical mastitis: farms in the upper three quartiles of prevalence were used as occurrence points for the Ecological niche modelling procedure ('infected farms'); farms in the lower quartile of prevalence within each category were (pseudo)negative points. Significant differences in environmental parametres prevailing in locations of farms into the study, were identified for up to 13 parametres between locations of farms according to management system applied in farms. When farms in each management system were considered separately, differences became evident between farms in each management system, as well as between the two infections. The factor with the highest relative contribution in the analyses was the distance from other sheep farms; other factors also of importance in the predictive models were the altitude, the maximum temperature of warmest month and the total precipitation of driest month. Verification of the model revealed that ≥ 0.760 of infected farms' were located in areas predicted as high risk for prevalence of subclinical mastitis or slime staphylococcal subclinical mastitis. The paper describes for the first time potential association of mastitis with environmental factors and presents predictive models for mastitis in ewes taking into account environmental parametres.

Experimental study for evaluation of the efficacy of a biofilm-embedded bacteria-based vaccine against Staphylococcus chromogenes-associated mastitis in sheep.

Although coagulase-negative staphylococci are the primary aetiological agents of subclinical mastitis in ewes, there is little information regarding vaccination against that infection. The objective of this study was to evaluate the efficacy of a vaccine against staphylococcal mastitis in ewes under experimental conditions. The antigen in the vaccine is based on a bacterin of Staphylococcus aureus strain, expressing the exopolysaccharide poly-N-acetylglucosamine (PNAG), which is involved in biofilm formation by these bacteria. Ewes in groups A (n = 17) or B (n = 6) were given an initial vaccination 5 weeks before expected lambing, followed by a repeat administration 21 days later. Ewes in groups C (n = 8) or D (n = 6) were unvaccinated controls. Ewes in group A (n = 17) or C (n = 8) were challenged with a biofilm-forming S. chromogenes; animals in subgroups A1 or C1 were challenged on the 10th and those in A2 or C2 on the 50th day after lambing. Ewes in groups B or D were uninoculated controls. Clinical examinations of ewes, ultrasonographic examinations of udder, milk yield measurements, blood sampling for detection of anti-PNAG specific antibodies and milk sample collection for bacteriological and cytological examinations were performed up to 52nd day post-challenge. Finally, biopsies were performed for mammary tissue collection for histopathological examination. Among group A ewes, 29% developed systemic signs and 59% signs in the inoculated gland; the respective figures for group C were 50% and 100% (P =  0.040 for mammary signs). The median total clinical score was 2.0 for A and 5.5 for C ewes (P =  0.025). For A, but not for C, clinical scores decreased progressively during the study (P =  0.018 and P =  0.47, respectively). The duration of mastitis was shorter in A (4 days) than in C (17.5 days) ewes (P =  0.022). Bacterial counts were lower in milk samples from A than from C ewes, for samples collected from the inoculated and the uninoculated (P <  0.01) mammary glands of these ewes. Somatic cell counts in samples from inoculated and uninoculated mammary glands of A ewes were higher than in samples of C ewes (P <  0.02). There were differences for gray-scale evaluations during ultrasonographic examination and for milk yield measurements between groups (P <  0.01). Median bacterial counts in tissue samples from A ewes (0 cfu g-1) were lower than in ones from C (6.5 cfu g-1) ewes (P =  0.041). The median score for histopathological findings in tissue samples from inoculated glands of A was lower than that for C ewes: 1 versus 2 (P =  0.014). It is concluded that mastitis was less severe in vaccinated animals, as indicated by a wide array of measures.

Animal-based measurements of the severity of mastitis in dairy cows.

A range of clinical parameters were studied to assess their usefulness as objective markers of the severity of clinical mastitis in dairy cows. Cows with moderate clinical mastitis had significantly higher rectal temperatures and heart and respiratory rates than cows with mild clinical mastitis or control cows. The difference in temperature between quarters did not vary significantly between the control cows and the cows with mastitis, but there was a larger difference between quarter temperatures in the cows with moderate mastitis than in the cows with mild mastitis or the control cows. The hock-to-hock distance in the control cows was significantly smaller than in the cows with mild to moderate mastitis, but there was no significant difference in the distance either between the cows with mild mastitis and those with moderate mastitis, or between the cows with mastitis in the front quarters and those with mastitis in the hind quarters. The mechanical threshold to pain of the cows with mild and moderate mastitis was significantly lower than that of the control cows.

Evaluation of an ELISA to assess the intensity of Fasciola hepatica infection in cattle.

An elisa with a diagnostic sensitivity of 98 per cent and specificity of 96 per cent was evaluated as a means of assessing the intensity of Fasciola hepatica infection in cattle. A total of 294 blood samples were collected from infected cattle at a local abattoir, and the level of infection in each animal was assessed on the basis of the extent of liver pathology and the presence of flukes; 120 blood samples were also collected from uninfected cattle kept on a farm known to be free of F hepatica. The results indicated that there was a significant correlation (P<0.001) between the elisa values and the intensity of infection. Values between 15 and 28 per cent of a positive control sample indicated a low intensity of infection, values between 28 and 50 per cent indicated a medium intensity of infection and values above 50 per cent indicated a high intensity of infection.

Serum cardiac troponin I concentrations in dogs with generalised seizures.

OBJECTIVES: To determine if serum cardiac troponin I concentrations - measured with both a first-generation assay and a high-sensitivity assay - were greater in dogs with generalised seizures than in controls and to identify clinical variables associated with cardiac troponin I concentration. MATERIALS AND METHODS: Prospective study of 30 dogs with recent generalised seizures and 30 healthy controls. Serum cardiac troponin I concentration was measured using two commercially available assays, and the correlation of clinical factors with concentration was examined. RESULTS: Serum concentrations of cardiac troponin I were higher in dogs that had recent seizures compared to controls when measured by both assays. The predictors most clearly associated with cardiac troponin I concentration were number of seizures and age. Both predictors were positively associated with increasing concentrations of troponin I. CLINICAL SIGNIFICANCE: Serum cardiac troponin I concentration was significantly elevated in dogs that had recent generalised seizures when compared to controls, and concentrations were higher in dogs that experienced more seizures. This association may indicate that generalised seizures are associated with damage to the myocardium.

Neospora caninum in cattle: experimental infection with oocysts can result in exogenous transplacental infection, but not endogenous transplacental infection in the subsequent pregnancy.

Whilst it is presumed that infection of pregnant cattle with Neospora caninum oocysts can provoke abortion and is the likely cause of epidemic abortion outbreaks, only two previous experiments have involved inoculation of pregnant cows with oocysts (and only one abortion was provoked in 22 pregnancies). Here, we describe the oral oocyst challenge of 18 cows synchronously bred and inoculated precisely at 70 (n=6), 120 (n=6) and 210 (n=6) days in pregnancy with a nominal dose of 40,000 oocysts. Only one abortion occurred (at the 120 days challenge) which could be definitively ascribed to N. caninum and no transplacental infection (TPI) was detected in any of the other 11 calves born in the 70 and 120 day challenge groups. In contrast, 4/5 live calves born to cattle challenged at 210 days were transplacentally infected. When cows which had transplacentally infected their calves in the first pregnancy were rebred, no TPI occurred. The results show that the timing of challenge influences clinical and parasitological outcomes and that cattle in late pregnancy are exquisitely sensitive to oocyst challenge leading to exogenous TPI and congenitally infected calves. However, cattle which were indisputably systemically infected in their first pregnancy did not induce endogenous TPI in their subsequent pregnancy. This confirms previous results with experimental tachyzoite challenge and suggests that post-natal infection does not lead to persisting infections which can recrudesce in pregnancy.

Evaluation of an enzyme-linked immunosorbent assay for detection of antibodies to Fasciola hepatica in milk.

Antibodies against Fasciola hepatica were detected in serum and individual milk samples of dairy cattle using an ELISA. Percentage positivity (PP) values in milk samples were related to serum PP values and were not influenced by days into lactation. The correlation coefficient between serum and individual milk samples was highly significant (r=0.84, P<0.005). The correlation coefficient between herd seroprevalence and herd milk antibody prevalence was 0.96. The correlation coefficient between prevalence measured by faecal egg count and both seroprevalence and milk antibody prevalence within the herd was 0.87. The diagnostic sensitivity and specificity for milk were 92% (95% CI=89-96) and 88% (95% CI=85-91), respectively, when the serum test was considered as a gold standard. In conclusion, the level of antibody to F. hepatica in milk is significantly correlated with the antibody level in serum and this ELISA is suitable as a means of routine veterinary diagnosis of exposure to F. hepatica in cattle and an alternative to testing sera.

Bacterial flora and risk of infection of the ovine teat duct and mammary gland throughout lactation.

We collected samples of teat duct material and mammary secretion from ewes in three farms (flock A, polyparous n=7; flock B, polyparous n=6, primiparous n=4; flock C, polyparous n=4): 14 samples immediately after lambing (before sucking of lambs), 244 samples during the suckling period and 156 samples during the milking period. Conventional bacteriological techniques were used. The results were modeled using survival analysis, initially by the Kaplan-Meier method and then by the Cox Proportional Hazards method. Then, we calculated the minimum true risk of an "at-risk" teat or mammary gland being infected and analyzed these data with STATA using the GLLAMM program for Generalised Linear Latent and Mixed Models. During the suckling period, bacteria were isolated from 52 (21%) duct material and 19 (8%) secretion samples; respective results for the milking period were 20 (13%) and 9 (6%). There was an increased risk of duct rather than secretion samples being infected (P<0.001). There was a significant difference among flocks in isolating bacteria from duct (P<0.01) or secretion (P<0.001) samples during suckling period, but not during hand-milking period (P>0.4 and 0.1, respectively). There were no differences between isolation of bacteria from duct (P>0.5) or secretion (P>0.7) samples among primiparous and polyparous animals. Most bacterial isolates were staphylococci. Persistent isolation of the same bacterial species from duct material samples obtained from a particular ewe was recorded with five Staphylococcus spp. and two Mannheimia haemolytica isolates. The results indicate that infections of the teat duct can take place easily; however, not all infections result to infection of the mammary gland. The results support experimental evidence that defence mechanisms of the healthy teat are able to limit the infection. Maintenance of healthy teats contributes to effective defence mechanisms, and coupled with minimal infections of the teat duct, would contribute to the prevention of mastitis in ewes.

Teat lesions predispose to invasion of the ovine mammary gland by Mannheimia haemolytica.

Teat lesions, produced in ewes by an experimental chapping procedure, were found to facilitate experimental infection with Mannheimia haemolytica, as assessed by observations on infection of the teat skin, teat duct and mammary gland, and on the production of mastitis. The origin of the M. haemolytica strain used (ovine tonsillar or mammary infection) did not appear to influence the results. In a second experiment, in which ewes continued to suckle their lambs but were not deliberately infected, chapping was shown to favour infection by Staphylococcus epidermidis, Staphylococcus aureus and M. haemolytica.