Discrepancies between the isolation of Salmonella from mesenteric lymph nodes and the results of serological screening in slaughter pigs

Most Salmonella control programmes are based on serological testing in the slaughterhouse. However, from a point of view of carcass contamination, it is rather the presence of Salmonella spp. in the animal at the time of slaughter that is important. The aim of this cross-sectional study was to investigate the possible discrepancies between the isolation of Salmonella spp. in the mesenteric lymph nodes and the results of serological screening. In total, 1821 fattening pigs originating from 60 Belgian farrow-to-finish herds were sampled in the slaughterhouse. The serum samples were analysed using an indirect mix-ELISA for the presence of Salmonella antibodies and evaluated at 3 cut-off values namely 10, 20, and 40% Optical Density (OD). All mesenteric lymph node samples were submitted to qualitative Salmonella isolation and a representative number of isolates was serotyped. From each herd, 30 animals were screened both serologically and bacteriologically and the herd was considered as positive when at least one sample was positive. At the herd level, 83.6% (cut-off OD 40%) to 100.0% (cut-off OD 10%) of the herds from which Salmonella had been isolated were evaluated as seropositive. At the individual level, only 34.5% (cut-off OD 40%) to 82.8% (cut-off OD 10%) of the animals from which Salmonella had been isolated were seropositive. Overall, a weak agreement was found between bacteriology and serology for Salmonella diagnosis. If pig herds are categorised using serological tests in the slaughterhouse, one should be aware of the fact that slaughter pigs can still harbour Salmonella spp. in the mesenteric lymph nodes, without being detected in serological tests. The cut-off value used to evaluate a sample as serologically positive and the number of samples per herd are of major importance to classify herds correctly in order to protect human health.     

Evaluation of vaccination with a commercial subunit vaccine on shedding of Salmonella enterica in subclinically infected dairy cows

OBJECTIVE: To estimate the efficacy of a commercially available Salmonella enterica subunit vaccine on the subclinical shedding of S enterica in dairy cattle. DESIGN: Randomized, controlled trial. ANIMALS: 175 mature cows on 2 dairy farms with a history of S enterica infection. PROCEDURES: 25% of the mature cows from each herd were systematically randomized to receive an S enterica subunit vaccine following label guidelines. The remaining 75% of cows in each herd served as nonvaccinated controls. Fecal samples were collected from all cows at the time of initial vaccination (day 0), booster vaccination (day 14), 2 weeks following the booster vaccination (day 28), and 10 weeks following the start of the trial (day 70). All samples were processed on the day of collection and cultured for S enterica. RESULTS: 651 fecal samples were obtained over the entire study period. Salmonella enterica was recovered from 46 (7.1%) of the samples. Shedding of S enterica was similar for vaccinated and nonvaccinated control cows on each of the collection dates. CONCLUSIONS AND CLINICAL RELEVANCE: The study revealed no evidence that extralabel vaccination with a commercial subunit S enterica vaccine reduced shedding of S enterica in subclinically infected dairy cows in these herds.     

Effect of organic acids in drinking water during the last 2 weeks prior to slaughter on Salmonella shedding by slaughter pigs and contamination of carcasses

In this study, we investigated the effect of adding organic acids to the drinking water of finishing pigs 2 weeks prior to slaughter on the shedding and prevalence rate of Salmonella at slaughter. Approximately 600 animals from four Belgian pig herds infected with Salmonella were included. At two herds, the study was conducted twice. Before the start of the study, overshoes were taken at the different herds. Two weeks prior to the expected slaughter date, the pigs were randomly divided into two groups (treatment and control group) each containing on average 50 animals within each herd. The treatment group received from this day onwards acidified drinking water (pH = 3.6-4.0), the control group received non-treated water (pH = 7.8-8.5). All other housing, feeding and management factors were identical in both groups. At the slaughterhouse, 10 pigs of each group (20 pigs for each group of study group 6) were randomly selected and sampled (blood, contents of ileum and rectum, mesenteric lymph nodes and carcass swabs). All samples were immediately transported to the laboratory and submitted to Salmonella isolation. Salmonella was isolated out of 11.9% (66/554) of the samples taken at the slaughterhouse, with the highest frequency found in the content of the ileum (18.7%), followed by 17.8% in the lymph nodes, 7.2% in the content of the rectum and 3.6% in the carcass swabs. The results did not reveal a significant difference between the treatment and control groups for the different slaughterhouse samples. The study documented that the investigated control strategy namely, the strategic application of organic acids during the last 2 weeks prior to slaughter was insufficient to decrease Salmonella shedding and contamination shortly before and during slaughter.     

The correlation between Salmonella serology and isolation of Salmonella in Danish pigs at slaughter

In Denmark, a serological Salmonella surveillance programme in finishing pig herds has been in place since 1995. The programme was founded on data from experimental studies, which demonstrated a strong association between Salmonella serology and the prevalence of these bacteria. The current study was carried out in three Danish abattoirs to evaluate the correlation under field conditions. A total of 160 Danish finishing pig herds were included. Seven out of these were examined twice, yielding a total of 167 observations. The herds were selected according to their herd serology based on data from the national surveillance. From each herd, samples were taken from 10 finishers at slaughter. The prevalence of Salmonella bacteria was measured at four sites: (1) caecal-content; (2) carcass surface; (3) pharynx; and (4) caecal lymph nodes. A logistic regression model was constructed for each sampling site. Abattoir, sanitary slaughter and herd seroprevalence were used as explanatory variables. The results demonstrated that there was a strong association between herd serology and the prevalence of Salmonella bacteria measured at three of the sampling sites: caecal-content, pharynx, and carcass surface. For these sites, the odds for being culture-positive for Salmonella varied from 1.3 to 1.5 for each increase of 10% in herd serology (P < 0.0001). For caecal lymph nodes, however, no linear association was found.     

Data-quality issues and alternative variable-screening methods in a questionnaire-based study on subclinical Salmonella enterica infection in Danish pig herds

Our aim was to determine risk factors for subclinical Salmonella enterica infection in Danish finishing-pig herds. In this paper, the evaluation, combining and initial reduction of variables is presented, along with assessment of the hypotheses in the preliminary statistical testing.The first group of herds was selected at random with no former knowledge of S. enterica infection. Both the herd prevalence and the within-herd prevalence among these herds turned out to be low; hence, some additional herds were selected from The Danish Salmonella Control program, based on their high seroprevalence. This resulted in a hybrid case-"control" design of the study and therefore, five different methods of categorising the data were used to ensure that variables were not wrongfully excluded as a result of using an improper design.Our questionnaire focused on management, infection-limiting precautions and feed and feeding procedures. To establish the prevalence of S. enterica infection within herds at the time of the visit, 50 blood samples from each herd were collected and serologically examined. The reliability of each variable from the questionnaire was assessed and it was decided which variables should be selected, disregarded, combined with other variables and/or recoded. In the simple statistical testing (2x2 tables, cut-off: P=0.25) herds were defined as subclinically S. enterica infected if the within-herd proportion of individual pigs with OD%>10 was more than 20%.The results included questionnaires from 96 randomly selected and 39 high-seroprevalence herds and 6814 blood samples. The initial 95 variables originally included in the questionnaires were reduced to 21 by critical check, combination, recoding and preliminary screening. We failed to demonstrate "herd size" as a risk factor for subclinical S. enterica infection in pig herds.     

Risk factors for the herd-level bacteriologic prevalence of Salmonella in Belgian slaughter pigs

Herd-level risk factors for salmonellosis in pigs were investigated in a cross-sectional study on 62 Belgian farrow-to-finish pig herds belonging to one slaughterhouse cooperative. Data concerning housing and ventilation, management, hygiene, biosecurity, production parameters, feeding, disease control and transport to the slaughterhouse were collected during a herd visit by means of a questionnaire. The percentage of positive animals in a slaughterhouse delivery, as determined by qualitative Salmonella isolation in the mesenteric lymph nodes taken from 30 slaughter pigs, was the outcome variable. All samples were taken in 4 different slaughterhouses. Variables first were submitted to a univariable analysis using a logistic mixed regression model, with herd as random effect. Variables which were related to the Salmonella prevalence (P < 0.05) were analysed further in a multivariable model. The clustering of Salmonella infection within a pen also was studied in a generalised mixed model with pen as random effect. Salmonella isolates were identified by serotype. In 57 (92%) of the herds, at least one sample was found positive for Salmonella. The median percentage of positive Salmonella samples per delivery was 64% (range: 0-100%). In the multivariable model, only type of floor was related significantly to the prevalence: 100% (95% CI 88-100) for herds with <50% slatted floors to 54% (36-70) for herds with fully slatted floors. The results from the analysis should be interpreted with care because only 62 herds were included in the study. Clustering between pigs from the same pen could not be demonstrated (variance +/- S.D.: 0.11 +/- 0.16). S. typhimurium (30%) and S. derby (20%) were most common among the 23 different serotypes that were found.     

Effect of acidified feed on the prevalence of Salmonella in market-age pigs

Two trials were carried out to determine the effect of feed acidification upon Salmonella carriage in market-age pigs. In the first trial, the administration for the last 14 weeks of the fattening period of a commercial pelleted feed added with 0.6% lactic acid plus 0.6% formic acid (Lac-Formic-1.2) was compared to an unacidified standard diet (STD). A second experiment was carried out in two herds of growing pigs (Herd I, 3000 pigs; Herd II, 900 pigs) in which three different diets were assayed during the last 8-9 weeks of the fattening period: a diet containing 0.8% formic acid (Formic-0.8), a diet containing 0.4% lactic acid plus 0.4% formic acid (Lac-Formic-0.8) and a STD. In the first experiment, serological evolution of the infection was examined by ELISA and microbiological cultures (rectal samples and mesenteric lymph nodes) were also done. Feed intake by pen and the individual weight of the animals were also measured. In the second trial, blood, rectal samples and mesenteric lymph nodes were collected at slaughter in both herds (30 pigs per experimental group). In the first experiment, the acidified diet (Lac-Formic-1.2) reduced Salmonella carriers in mesenteric lymph nodes (Fisher's exact P < 0.01). In the second trial, Lac-Formic-0.8 diet significantly reduced Salmonella seroprevalence compared to the STD (P = 0.001) in both herds. Also Lac-Formic-0.8 and Formic-0.8 diets in Herd II showed a lower faecal excretion and Salmonella carriage in mesenteric lymph nodes than the STD (P < 0.05). Our results suggest that the administration of a combination of lactic and formic acids at the levels used in this study could be used to reduce Salmonella prevalence in finishing pigs.     

Hepatitis E virus infection dynamics and organic distribution in naturally infected pigs in a farrow-to-finish farm

The objective of the present study was to determine the pattern of Hepatitis E virus (HEV) infection in a naturally infected, farrow-to-finish herd. For that purpose, a prospective study was conducted in randomly selected 19 sows and 45 piglets. Blood samples were collected from sows at 1 week post-farrowing and from piglets at 1, 3, 6, 9, 12, 15, 18 and 22 weeks of age. Furthermore 3 or 5 animals were necropsied at each bleeding day (but at 1 week of age), and serum, bile, liver, mesenteric lymph nodes and faeces taken. HEV IgG, IgM and IgA antibodies were determined in serum and viral RNA was analysed in all collected samples by semi-nested RT-PCR. Histopathological examination of mesenteric lymph nodes and liver was also conducted. From 13 analysed sows, 10 (76.9%) were positive to IgG, one to IgA (7.7%) and two to IgM (15.4%) antibodies specific to HEV. In piglets, IgG and IgA maternal antibodies lasted until 9 and 3 weeks of age, respectively. IgG seroconversion occurred by 15 weeks of age while IgM and IgA at 12. On individual basis, IgG was detectable until the end of the study while IgM and IgA antibody duration was of 4-7 weeks. HEV RNA was detected in serum at all analysed ages with the highest prevalence at 15 weeks of age. HEV was detected in faeces and lymph nodes for the first time at 9 weeks of age and peaked at 12 and 15 weeks of age. This peak coincided with the occurrence of hepatitis as well as with HEV detection in bile, liver, mesenteric lymph nodes and faeces, and also with highest IgG and IgM OD values at 15 weeks. Finally, different HEV sequences from this farm were obtained, which they clustered within 3 different groups, together with other Spanish sequences, all of them of genotype 3. Moreover, the present study also indicates that the same pig can be infected with at least two different strains of HEV during its productive life. This is the first study characterizing HEV infection in naturally infected pigs with chronological virus detection and its relationship with tissue lesions throughout the productive life of the animals.     

Seroprevalence and antibiotic sensitivity of serotypes of Salmonella enterica in Greek pig herds

Blood samples were taken from 50 pigs in each of 59 farrow-to-finish production herds and from 40 pigs in each of four of five registered multiplying herds. Samples of feed and faeces were also collected from 17 of the production herds and from the four multiplying herds. The sera were tested for antibodies to Salmonella enterica by the Danish mix-ELISA, and the organisms were isolated, serotyped and sensitivity tested by standard techniques. The average within-herd seroprevalence was 3.4 per cent and at least one pig tested seropositive in 21 of the 59 herds. In the multiplying herds, only a single seroreactor was detected. Salmonellae were isolated from only five of 95 feed samples, from two of the 17 herds sampled, Salmonella tennessee in four of five samples from one herd and an untypable strain in one of five samples from another. Four infected faecal samples were detected in four herds; they harboured Salmonella typhimurium, Salmonella bredeney or Salmonella london. No salmonellae were isolated from the samples of feed and faeces taken from the multiplying herds. The S london and S typhimurium had a low sensitivity to streptomycin, kanamycin and neomycin, and the S typhimurium also had low sensitivity to amoxycillin, ticarcillin, piperacillin, amoxycillin + clavulanic acid, cefalotin and cefoperazone. The other isolates were sensitive to all the antimicrobial agents tested.     

Effect of vitamin E supplementation on lymphocyte distribution in gut-associated lymphoid tissues obtained from weaned piglets

Fifteen piglets were used to determine the effect of vitamin E supplementation on the number of CD4-immunoreactive (CD4+) T-lymphocytes, CD8-immunoreactive (CD8+) T-lymphocytes and IgA-immunoreactive (IgA+) B-lymphocytes per follicle in the Peyer's patch of distal ileum and the mesenteric lymph nodes of weaned piglets. Piglets, following a 3-day adaptation period after weaning, were assigned to one of three experimental groups: control (no vitamin E supplementation), vitamin E supplementation of 100 mg/kg of diet and vitamin E supplementation of 300 mg/kg of diet. Supplementation of vitamin E lasted for a period of 36 days. The basal diet contained 80 mg alpha-tocopherol/kg of diet. All piglets were killed at day 39 after weaning and samples of the distal ileum and adjacent mesenteric lymph nodes were collected. The number of cells for each lymphocyte subset was counted in the Peyer's patch and the mesenteric lymph nodes follicles, in cryostat sections processed for immunohistochemistry. Results showed that vitamin E supplementation (300 mg/kg diet) of piglets caused an increase (P < 0.05) in the number of IgA+ B-lymphocytes in the Peyer's patch, but not in the mesenteric lymph nodes, compared with the corresponding values in control animals. Vitamin E supplementation had no effect (P > 0.05) on the number of CD4+ and CD8+ T-lymphocytes in the follicles of the Peyer's patch and the adjacent mesenteric lymph nodes. Thus, vitamin E had relatively minor effects on distribution of the major immunocompetent cells in the gut. The numbers of CD4+ and CD8+ T-lymphocytes as well as IgA+ B-lymphocytes per follicle were higher by 26-77% (P < 0.05) in the mesenteric lymph nodes than the corresponding values in the Peyer's patch.     

Herd-level risk factors for Salmonella enterica subsp. enterica in U.S. market pigs

Midwest U.S. herds (n = 63) were studied to identify risk factors for harboring Salmonella enterica among slaughter-weight pigs. Samples collected on farms (feces) and at slaughter (distal colonic content, cecal content and ileocolic lymph nodes) were cultured using conventional means. Approximately 15 pigs were studied per herd, for a total of 3754 samples. The proportion of pigs positive in one or more samples was calculated for each herd. Herd characteristics were described by a combination of interview and written survey. Logistic regression was used to detect relationships between the detection of Salmonella and potential herd-level risk factors. The mean individual pig prevalence was 5% for feces, 4% for distal colonic content, 15% for ileocolic lymph nodes, and 17% for cecal contents. One or more Salmonella isolates were detected in at least one sample type in every herd. The five most common serovars were S. Agona, S. Derby, S. Schwarzengrund, S. Typhimurium and S. Senftenberg, with 25 additional serovars detected. Salmonella prevalence estimates were positively correlated among all samples except distal colonic content and ileocolic lymph nodes. Pigs with culture positive fecal samples were at increased odds of being detected positive for each of the slaughter-collected samples examined, namely distal colonic content (OR = 30.5), ileocolic lymph nodes (OR = 12.9) and cecal content (OR = 23.2). Herds with positive fecal sample(s) had increased odds of having positive cecal content (OR > 1.5), distal colonic content (OR = 15.3) and ileocolic lymph nodes (OR = 12.7). Pigs from herds with at least some bowl drinkers had eight-fold higher odds of testing Salmonella positive than did pigs from herds with only nipple drinkers. Pigs from herds with only dry feeders had five-fold higher odds of testing Salmonella positive when compared with pigs from herds with combinations of wet/dry style feeders. Interventions at these two points should be considered when designing growing pig facilities to reduce Salmonella shedding.     

The emergence of Mycobacterium paratuberculosis in farmed deer in New Zealand - a review of 619 cases

AIM: To review cases in which Mycobacterium paratuberculosis was identified in farmed deer in New Zealand. METHODS: Case histories were reviewed where M. paratuberculosis was identified in deer by either culture or a polymerase chain reaction (PCR) test using primers from IS900. RESULTS: Between 1986 and 2000, M. paratuberculosis was identified by bacterial culture and/or PCR in 619 farmed deer from 299 herds, representing approximately 6% of deer herds in New Zealand. Over 85% of cases were identified during the last 6 years. In 60% of the infected herds, only one infected animal was identified. The maximum number of cases identified in a single deer herd was 47, and these were identified over a period of 8 years. Only 36 (5.8%) cases came from clinically affected animals identified on farms by veterinarians. The majority (89.7%) of the 619 cases were identified from lesions in mesenteric lymph nodes, including the ileocaecal lymph nodes, identified at meat inspection as being macroscopically either typical or equivocal of bovine tuberculosis (M. bovis). While the overwhelming majority of lesions were identified in mesenteric lymph nodes, M. paratuberculosis was also identified in 27 lesions in lymph nodes of the head, especially the retropharyngeal lymph node. CONCLUSIONS: The figures presented underestimate the true prevalence of infection with M. paratuberculosis, especially since not all suspect cases were submitted for culture or PCR. However, they do show that M. paratuberculosis appears to be spreading in farmed deer in New Zealand and highlight the possibility that Johne's disease is emerging as a potential major problem affecting this species. Identification of the organism by bacterial culture or PCR is required in many cases to distinguish lesions in mesenteric lymph nodes and lymph nodes of the head caused by M. paratuberculosis from those caused by M. bovis and M. avium.     

Prevalence of Salmonella serotypes on pig carcasses from high- and low-risk herds slaughtered in three abattoirs

The aim of this study was to compare the prevalence of Salmonella serotypes at two different sites on pig carcasses from herds classified as high-risk or low-risk and to elucidate the relationship between carcass contamination levels and serological status. Caecal samples and carcass surface swabs were cultured for Salmonella from a total of 210 pigs from low risk herds (< 19% of pigs in herd Salmonella seropositive) and 209 pigs from high risk herds (> 32% of pigs in herd Salmonella seropositive) in three abattoirs. Meat juice samples were collected for analysis by ELISA. The prevalence of Salmonella in the caecal contents of "low-risk" pigs was 10%, which was significantly lower than the 19% prevalence in "high-risk" pigs (p < 0.01). The corresponding figures for skin samples collected immediately post-evisceration were 2% and 12%. The predominant Salmonella serotype in the caecal contents of both the low-risk and high-risk pigs was Salmonella Typhimurium. Salmonella Kentucky and Salmonella Derby were the most frequent isolates from the carcass surface swabs of low- and high-risk pigs respectively. There was a positive association between seropositivity of pigs from high-risk herds and caecal carriage (p < 0.05). Results showed that herd categorisation based on serological results was useful in predicting Salmonella isolation rates from caecal samples and surface swabs of slaughtered pigs.     

Simulation model estimates of test accuracy and predictive values for the Danish Salmonella surveillance program in dairy herds

The Danish government and cattle industry instituted a Salmonella surveillance program in October 2002 to help reduce Salmonella enterica subsp. enterica serotype Dublin (S. Dublin) infections. All dairy herds are tested by measuring antibodies in bulk tank milk at 3-month intervals. The program is based on a well-established ELISA, but the overall test program accuracy and misclassification was not previously investigated. We developed a model to simulate repeated bulk tank milk antibody measurements for dairy herds conditional on true infection status. The distributions of bulk tank milk antibody measurements for infected and noninfected herds were determined from field study data. Herd infection was defined as having either >or=1 Salmonella culture-positive fecal sample or >or=5% within-herd prevalence based on antibody measurements in serum or milk from individual animals. No distinction was made between Dublin and other Salmonella serotypes which cross-react in the ELISA. The simulation model was used to estimate the accuracy of herd classification for true herd-level prevalence values ranging from 0.02 to 0.5. Test program sensitivity was 0.95 across the range of prevalence values evaluated. Specificity was inversely related to prevalence and ranged from 0.83 to 0.98. For a true herd-level infection prevalence of 15%, the estimate for specificity (Sp) was 0.96. Also at the 15% herd-level prevalence, approximately 99% of herds classified as negative in the program would be truly noninfected and 80% of herds classified as positive would be infected. The predictive values were consistent with the primary goal of the surveillance program which was to have confidence that herds classified negative would be free of Salmonella infection.     

Isolation of Salmonella organisms from the mesenteric lymph nodes of horses at necropsy.

OBJECTIVES: To determine the prevalence of Salmonella infections in horses at necropsy. DESIGN: Cross-sectional prevalence survey. ANIMALS: 102 horses. PROCEDURE: Mesenteric lymph nodes were collected from horses that were necropsied. Horses had died or were euthanatized because of severe disease or at the request of the owner. Twenty-eight of the horses were racehorses euthantized following acute catastrophic injuries on the racetrack. Mesenteric lymph nodes were submitted for Salmonella culture via direct plating of tissue specimens on MacConkey agar and by use of 4 enrichment culture techniques that used tetrathionate and selenite enrichment broth and brilliant green and Salmonella-Shigella selective plating media. RESULTS: Salmonella typhimurium was isolated from the mesenteric lymph nodes of 2 foals (2/102, 1.96% of the horses). Salmonella organisms were not isolated from the mesenteric lymph nodes of adult horses. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of Salmonella infections in horses of our study (1.96%) suggests that the results of cross-sectional surveys, using bacteriologic culture to determine prevalence of Salmonella infection, should be interpreted with caution. Prevalence of Salmonella infections determined in a single facility may not reflect the prevalence of Salmonella-infected horses in the general population; furthermore, obtaining a Salmonella isolate from a horse does not establish that the horse is a chronic Salmonella carrier.     

Prevalence of fecal shedding of Salmonella spp in dairy herds

OBJECTIVE: To estimate prevalence of Salmonella spp in Ohio dairy farms and to identify potential risk factors for fecal shedding of salmonellae. DESIGN: Cross-sectional study. SAMPLE POPULATION: 105 Ohio dairy farms. PROCEDURE: Individual fecal samples from all mature cows in study herds were tested for Salmonella spp by use of standard bacteriologic culture procedures. Herds were identified as infected if at least 1 cow was shedding Salmonella spp. Information regarding herd characteristics, management practices, and health history were collected. Potential risk factors for herd-level Salmonella infection were identified. RESULTS: In 31% of the study herds (95% confidence interval, 22 to 40%), at least 1 cow was shedding Salmonella spp. Six percent of 7,776 fecal samples contained Salmonella organisms; prevalence within infected herds ranged from < 1 to 97%. Herd size, use of free stalls for lactating and nonlactating cows, and use of straw bedding in nonlactating cows were significantly associated with fecal shedding of Salmonella spp, as determined by use of univariate analysis. By use of multivariate analysis, large herds were more likely to be infected than smaller herds; however, no other factors were associated with Salmonella infection after adjustment for herd size. CONCLUSIONS AND CLINICAL RELEVANCE: Subclinical shedding of Salmonella spp is common in Ohio dairy herds, although we could not identify specific interventions that may influence the prevalence of Salmonella spp on dairy farms. It appears that large herd size and intensive management may provide an environment conducive to Salmonella shedding and chronic dairy herd infection.     

Exudative epidermitis and porcine circovirus-2 infection in a Swedish SPF-herd

An outbreak of exudative epidermitis (EE) among piglets in a Swedish SPF-herd initiated a survey for indications as to the cause of disease.The herd was established by caesarean section and has been closed to all new animal material, with the exception of semen for artificial insemination (AI). The study comprised serum samples from the SPF-herd over a 10-year period (n=109) and a close monitoring of animals in the herd during the period after the EE outbreak. Serum samples from conventional boars at the AI-station servicing the herd were also included (n=9). All serum samples were tested for antibodies to porcine circovirus-2 (PCV-2). In addition, 3-week-old piglets from three litters (n=24) farrowed close after the initial EE outbreak were closely monitored for clinical signs of skin disease, sampled for Staphylococcus hyicus, tested for antibodies to porcine parvovirus and in sequentially collected serum samples tested for interferon-alpha (IFN-alpha) and interleukin-6.The PVC-2 serology showed that animals in the herd were sero-negative at least until 2 months prior to the EE outbreak. During the period close after the EE outbreak the animals showed varying levels of antibodies to PCV-2 but all the tested animals had sero-converted 4 months later. The AI boars were also sero-positive to PCV-2 at the time of the EE outbreak. Animals in the SPF-herd remained sero-positive to PCV-2 during the following 7 years. In the monitored litters, one piglet had clinical EE and 15 piglets displayed defined erythemas on the abdomen. Fourteen of the piglets also had IFN-alpha in serum on one or more occasions during the study, indicating viral activity among the animals. S. hyicus was isolated from all of the piglets from the earliest sampling point (3 days of age) and onwards, irrespective of clinical signs. PCV-2 was isolated from lymph node tissue collected from one of the EE affected pigs.Further, increases in the number of stillborn piglets, small litters (<6 piglets) and repeat breeders could be correlated to the time of PCV-2 sero-conversion. Coincidence of active viral infection and sero-conversion to PCV-2 points to the virus as the cause of the EE outbreak and reproductive disturbances.     

Transfer of humoral and cell-mediated immunity via colostrum in pigs

The first aim of our study was to obtain information on the transmission of antigen-specific antibodies from colostrum to respiratory tract mucosa in piglets. The second aim was to confirm the biological relevance of the presence of lymphocytes in colostrum and the already described fact that these cells can penetrate the intestinal barrier and "colonize" peripheral blood and lymphatic tissues of piglets. Therefore, we performed an experiment in which sows were immunized with a model antigen Keyhole Limpet Hemocyanin and their piglets were euthanized at different intervals after birth and colostrum intake. Colostrum, bronchoalveolar lavage fluid and blood samples were collected for serological detection of antigen-specific antibodies. Lymphocytes isolated from peripheral blood and lymphatic tissues (mesenteric and tracheobronchial lymph nodes and spleen) of piglets were in vitro activated with the antigen. We found that colostrum-derived antibodies can cross into the respiratory tract mucosa. Furthermore, we found that antigen-specific lymphocytes were detectable in mesenteric lymph nodes and peripheral blood, but very rarely in spleen and tracheobronchial lymph nodes.     

A hierarchical trend model for bovine virus diarrhoea virus (BVDV) sero-conversion in Norwegian dairy herds from 1993 through 1997

The Norwegian bovine virus diarrhoea (BVD) control-and-eradication program performed annual testing of the national cattle population since its first screening in January 1993. A bulk-tank milk ELISA antibody test was used for the initial screening of dairy herds. Based on the annual bulk-tank milk-test result, a binary variable denoting herd sero-conversion (a surrogate measure for incidence) was created.The count of herds with sero-conversion in each community was regressed on year and the initial herd-sero-prevalence for each community in a Poisson hierarchical trend model - modelling the risk of sero-conversion. By using this modelling approach, estimates of trend specific for each hierarchical level of organisation were included in the trend model (community, veterinary district and county) could be estimated.The main BVD trend showed a steadily declining sero-conversion risk. The communities in the highest herd-sero-prevalence quartile in 1993 continued to have the highest sero-conversion risk throughout the study period--decreasing from an average predicted sero-conversion risk in 1993 of 0.12 (95% CI; 0.10, 0.13) to 0.02 (0.007, 0.04) in 1997. There was an expressed variation in the level of sero-conversion for all the three hierarchical levels, but the trend only varied at the lowest level (community).     

Evaluation of herd sampling for Salmonella isolation on midwest and northeast US dairy farms

Epidemiologic investigations of Salmonella infections in dairy cattle often rely on testing fecal samples from individual animals or samples from other farm sources to determine herd infection status. The objectives of this project were to evaluate the effect of sampling frequency on Salmonella isolation and to compare Salmonella isolation and serogroup classification among sample sources on 12 US dairy farms sampled weekly for 7-8 weeks. Three herds per state were enrolled from Michigan, Minnesota, New York and Wisconsin based upon predefined herd-size criteria. Weekly samples were obtained from cattle, bulk tank milk, milk filters, water and feed sources and environmental sites. Samples were submitted to a central laboratory for isolation of Salmonella using standard laboratory procedures. The herd average number of cattle fecal samples collected ranged from 26 to 58 per week. Salmonella was isolated from 9.3% of 4049 fecal samples collected from cattle and 12.9% of 811 samples from other sources. Serogroup C1 was found in more than half of the samples and multiple serogroups were identified among isolates from the same samples and farms. The percentage of herd visits with at least one Salmonella isolate from cattle fecal samples increased with overall herd prevalence of fecal shedding. Only the three herds with an average fecal shedding prevalence of more than 15% had over 85% of weekly visits with at least one positive fecal sample. The prevalence of fecal shedding from different groups of cattle varied widely among herds showing that herds with infected cattle may be classified incorrectly if only one age group is tested. Testing environmental sample sources was more efficient for identifying infected premises than using individual cattle fecal samples.