Drag swab efficiency factors when sampling chicken manure.

This study examines drag swabbing distance, media for moistening the drag swabs, and site selection when sampling a laying facility by drag swabbing manure piles. Manure piles at a laying facility in California's San Joaquin Valley were sampled with drag swabs over various distances. Samples were cultured for Salmonella spp. with standard laboratory methods, and most probable number calculations. Salmonella spp. counts were expected to be highly variable because of reported clustering. Therefore, total bacteria and Escherichia coli, which were assumed to have a more uniform distribution on the surface of the manure, were additionally used as proxies for Salmonella. Media for moistening the swabs were compared by seeding postswabbing samples with Salmonella typhimurium, and culturing at different delay times. Total bacterial counts were compared between samples that were obtained from either wet or dry surfaces. Numbers of Salmonella spp. and total bacteria peaked within 120 feet of swabbing distance. Higher total bacteria counts were obtained by swabbing wet areas rather than dry areas, but the distance that could be swabbed effectively was shorter in wet areas. Moistening media selected for the swab resulted in statistically different culture counts, but did not show any important difference in maintaining Salmonella viability over a 48-hr period when the samples were kept at refrigerated temperatures. Once swabs became fully loaded with fecal material, bacterial numbers failed to increase with further use. Overuse of a swab may result in failure to detect Salmonella enteritidis on chicken manure if the distribution of this organism is clustered.

Decontaminating beef for Escherichia coli O157:H7.

Beef lean, fat, and connective tissues were inoculated with Escherichia coli O157:H7 before and after a prewashing procedure to compare the efficacy of prewashing and no prewashing on bacterial adherence and, consequently, on the removal of bacteria from the inoculated surfaces. Prewashing consisted of spraying tissues with tap water before inoculation. Final washing with disinfectant solutions compared the efficacy of several chemicals for the removal or destruction of E. coli O157:H7. The results showed that prewashing was very effective in reducing the numbers of bacterial cells on beef tissues, mainly lean tissue, in the control samples which received final washing with water. An opposite effect of prewashing was observed when disinfectant solutions were used for final washing; this may be due to dilution by water carried on the tissues after prewashing. The efficacy of chemicals was dependent on the type of exposed tissue. Hydrogen peroxide (3%) was more efficient in the removal of E. coli O157:H7 from connective tissues, with reductions greater than 4 log CFU/cm2, compared to a normally washed control (P < 0.01). Chlorhexidine (0.1%) was very efficient on fat and lean tissues, causing reductions over 5 log CFU/cm2 on not prewashed fat and lean tissues, compared to the control (P < 0.01). Acetic acid (5%) was the least effective, decreasing the number of CFU by under 1 log/cm2 as compared to the control; and no statistically significant difference was found among tissues, even though the removal of bacteria seemed less in lean tissue compared to fat or connective tissues.

Microbial food borne pathogens. Escherichia coli O157:H7.

Escherichia coli O157:H7 has emerged in the past 20 years as a pathogen of public health importance. Although most E. coli are normal flora in the colons of humans and other warm-blooded animals, several strains are capable of causing disease in humans. In recent years, E. coli O157:H7 and other shiga-like toxin-producing strains have been transmitted via foods and caused disease ranging from bloody diarrhea, and in more severe cases, hemolytic uremic syndrome and thrombocytopenic purpura. The reservoir appears to be cattle and, perhaps, other ruminants. Control is difficult in nonheated foods due to the organism's tolerance to low pH. Only supportive, symptomatic treatment is available for affected humans, and means to eliminate carriage in livestock are not presently available.

A sero-epidemiological study of Haemophilus somnus infection in dairy cattle.

Five repeated cross-sectional serological surveys of 790 dairy cattle in 4 dairy herds between December 1985 and February 1987 provided an opportunity to study the changes in the seroprevalence of Haemophilus somnus across the 5 surveys and with respect to some demographic and disease variables. The demographic variables included were age (heifers or cows) and farm, representing two groups of herds (two herds in each group, located in the Central and Northern Valleys of California). The serological status of cattle as either negative or positive against H. somnus, Campylobacter fetus and Leptospira hardjo were determined with enzyme linked-immunosorbent assays. Logistic regression analysis was used to compute maximum likelihood estimates of adjusted odds ratios and their respective 95% confidence intervals. The baseline risk of being H. somnus seropositive in the cattle observed at first sampling did not vary significantly during the study period after adjustment for the effects of covariates. Only at first sampling were cows about twice as likely to be H. somnus seropositive than heifers. At samplings 1 and 3, but not at 2 and 5, being in the herds of the Central Valley appeared protective. In contrast, at sampling 4 the cattle in herds in the Central Valley were about 7 times more likely to be H. somnus seropositive. C. fetus-positive cattle were about 3 times more likely to be H. somnus seropositive at sampling 1 only. The relationship between H. somnus status and L. hardjo was not significant during the study period.

Application of molecular biological techniques for detection of epizootic hemorrhagic disease virus (EHDV-318) recovered from a sentinel calf in central Sudan.

Epizootic hemorrhagic disease virus (EHDV), isolate 318 (EHDV-318), an untyped virus recovered from a sentinel calf herd at the Khartoum University farm in central Sudan, was characterized using molecular biological techniques. With dot blot hybridization technique, a cDNA probe derived from genome segment 6 of EHDV-2 (Alberta strain) hybridized with RNA from EHDV-318. Application of serogroup-specific EHDV polymerase chain reaction (PCR) to EHDV-318 RNA resulted in specific amplification of a 387 bp PCR product. Amplification product was visualized on ethidium bromide-stained agarose gel. Specificity of the PCR products was confirmed by chemiluminescent hybridization with a non-radiolabelled internal probe. No amplification product or hybridization signal was detected when the serotype-specific EHDV-1 or EHDV-2 PCR-based assays were applied to RNA from EHDV-318. The scientific data presented in this study indicated that cDNA probes and serogroup-specific PCR-based assay can be used to classify the virus as a member of EHDV serogroup, and as serotypically distinct from EHDV-1 and EHDV-2.

Application of enzyme-linked immunosorbent assay for epidemiological studies of diseases of livestock in the tropics of Mexico.

This study was conducted at the Centre for Research, Teaching and Extension in Tropical Livestock (Centro de Investigación, Enseñanza y Extensión en Ganadería Tropical) of the Faculty of Veterinary Medicine of the National Autonomous University of Mexico. During the latter part of 1986 and throughout 1988 and 1989, the herd of Holstein x zebu cattle at the University was tested for IgG antibodies to twenty-one viral, bacterial, rickettsial and parasitic agents. Antigens prepared from twenty infectious disease agents were used as the solid phase in an enzyme-linked immunosorbent assay, and the agar gel immunodiffusion procedure was used to test for antibodies against bovine leukaemia virus. The prevalence of IgG antibodies was high (> 50%) for bluetongue virus, Anaplasma marginale and Mycoplasma bovis. Antibodies to Brucella abortus were absent and antibodies against bovine virus diarrhoea virus and infectious bovine rhinotracheitis virus showed a very low prevalence (< 5%). Antibodies to fifteen other antigens showed intermediate prevalence (15-46%). Antibodies to Campylobacter fetus, A. marginale, bluetongue virus, bovine leukaemia virus and Haemophilus somnus displayed seasonal variations. Levels of antibody to bovine leukaemia virus, M. bovis and Listeria monocytogenes exhibited increasing secular trends while antibodies to bovine virus diarrhoea virus and C. fetus showed declining trends. Prevalence of antibodies increased with the age of animals tested. No consistent difference in antibody prevalence was found between three genotypic groups examined.

The association between antibody titres against Campylobacter fetus and reproductive efficiency in dairy cattle.

The relationship between the antibody titres against Campylobacter fetus and various indices of reproductive efficiency was studied in a cross-sectional study of 178 dairy cows from three California Dairy Herd Improvement Association herds. Blood samples were collected from the lactating cows during December 1986. Enzyme-linked immunosorbent assays were used to determine the antibody titres of the cow against Campylobacter fetus, Haemophilus somnus and Leptospira hardjo and were classified as either negative or positive. The status of a cow as either negative or positive against Campylobacter fetus and Haemophilus somnus represents serological evidence of natural exposure to the corresponding bacterial agents. However, the status against Leptospira hardjo was assumed to reflect a vaccinal titre since all the cows studied had been routinely vaccinated against this organism in September 1986. The data on demographic and reproductive parameters pertained only to the current lactation of the cows and were obtained from Dairy Herd Improvement Association individual cow records of December 1986. Five indices of reproductive efficiency were used, namely the recent calving interval, the calving-to-conception interval, the calving-to-last-service interval, the number of services per conception, and the number of services since last calving. The serological status against Haemophilus somnus, Leptospira hardjo and other covariates suggested by the results of previous studies were included in modelling the relationships of interest. Stepwise multiple linear regression analyses were carried out to study the adjusted relationship of Campylobacter fetus with each measure of reproductive efficiency. Multivariate analyses revealed that the adjusted relationship for Campylobacter fetus with all five measures of reproductive efficiency was non-significant (p > 0.05). Among the covariates, Leptospira hardjo had a strong and independent relationship with recent calving interval, the unstandardized partial regression coefficient being -0.77. The possible biological mechanisms of these associations are discussed.

The association between antibody titres against Campylobacter fetus and milk production efficiency in dairy cattle.

The association between serological evidence of exposure to Campylobacter fetus and milk production performance was studied in 178 lactating cows from three California Dairy Herd Improvement Association herds using a cross-sectional study design in December 1986. ELISAs were used to determine the antibody titres against Campylobacter fetus, Haemophilus somnus and Leptospira hardjo, which were classified as either negative or positive. The status of a cow as negative or positive against C. fetus and H. somnus represents the serological evidence of natural exposure to the corresponding bacteria. However, the status against L. hardjo was assumed to be the level of vaccinal titre against this organism since all the cows studied had been vaccinated against this agent. The data on demographic and productivity variables relating to the current lactation of the cows were obtained from Dairy Herd Improvement Association individual cow records for December 1986. Four measures of milk production efficiency for the current lactation were used. The status against L. hardjo and other covariates suggested by previous studies were included in modelling the relationships of interest. Stepwise multiple linear regression was used to study the adjusted relationship of C. fetus with each measure of milk production efficiency. Multivariate analyses revealed that the adjusted relationships of C. fetus with the test-day's milk production, the extended 305-day milk production and the relative value of milk production were not significant (p > 0.1). However, after adjusting for possible covariates, C. fetus-positive cows had an average of 7.43% lower mature equivalent milk production than C. fetus-negative cows (p = 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Mass screening of cattle sera against 14 infectious disease agents, using an ELISA system for monitoring health in livestock.

Mass screening ELISA methods were developed for testing cattle serum for antibodies against 14 common livestock diseases simultaneously. The absorbance values were transformed to a %ELISA (spectrophotometric antibody end point) by a computer interfaced with a microplate reader. A histogram indicating a cutoff point and a report for the veterinarian also was generated. The computer program produced a print-out of the antibody profile for each animal tested, the antibody concentration against each disease, and a histogram (antibody profile) showing the prevalence of each disease in the herd. Serum samples were obtained from 1,953 cattle, including 880 dairy cattle from 10 herds and 1,073 beef cattle from 20 herds. These samples were obtained from June 1988 through June 1989. The highest antibody prevalence was against bluetongue virus. Of the 1,953 cattle tested, 1,223 (63%) were seropositive for bluetongue virus, including 502 (57%) of the dairy cattle and 721 (67%) beef cattle. Other antibody prevalences, in descending order, were: rotavirus (44%), Pasteurella spp (25%), Leptospira spp and Haemophilus spp (22%), Mycoplasma spp (18%), parainfluenza virus (17%), Campylobacter spp (16%), Anaplasma marginale (15%), bovine leukosis virus (13%), Brucella spp (8%), Mycobacterium paratuberculosis (8%), bovine viral diarrhea virus (3%), and infectious bovine rhinotracheitis virus (3%). Major differences in antibody prevalence between dairy and beef cattle were that only 4% of the dairy cattle were seropositive for A marginale, compared with 25% of the beef cattle, and conversely, 29% of the dairy cattle were seropositive for bovine leukosis virus, compared with 1% of the beef cattle.(ABSTRACT TRUNCATED AT 250 WORDS)

The use of loglinear model to evaluate factors associated with sero-prevalence of Campylobacter fetus in dairy cattle.

Campylobacter fetus is one of the leading infections causing bovine infertility and abortion. In a cross-sectional study, the associations of parity, herd, serological status against Leptospira hardjo and Haemophilus somnus with the sero-prevalence of Campylobacter fetus in dairy cattle were evaluated. Four hundred dairy cows from three herds were included in the study. Blood samples from the cows were collected in December 1986. An enzyme-linked immunosorbant assay (ELISA) was used to determine positive or negative status of cows against Campylobacter fetus , Haemophilus somnus and Leptospira hardjo antigens. Loglinear analysis was used to describe the relationship of four categorical variables with the sero-prevalence of Campylobacter fetus . The final loglinear model was interpreted as a logit model. The final loglinear model included a three-way interaction among Campylobacter fetus , Leptospira hardjo and herd and a two-way interaction between Campylobacter fetus and parity. The adjusted odds of being a positive Campylobacter fetus reading were highest for cows with parity >6 (e.g. 27.8, 95% confidence interval (CI) = 12.9, 60.1); it was next highest for cows in parity <4 (e.g. 18.0, 95% CI = 13.5, 24.2); and it was lowest for cows in parity 4 to 6 (e.g. 13.3, 95% CI 7.3, 24.4). The herd management practice that was associated with the carrier state in early and late parities is identified in the study, and the biological implication of a three-way interaction among Campylobacter fetus , Leptospira hardjo and herd are discussed.

A cross-sectional study of bluetongue virus and Mycoplasma bovis infections in dairy cattle: I. The association between a positive antibody response and production efficiency.

In a cross-sectional study of bluetongue virus (BTV) and Mycoplasma bovis (M. bovis) infections, a sample of 572 California dairy cows was tested for the presence of antibodies to answer the question: Is it possible to identify and to assess quantitatively the associations between positive antibody test and production? Serum samples collected from these cows during December 1986 were tested for the presence of antibodies to BTV and M. bovis using the enzyme-linked immunosorbent assay (ELISA). Data on milk production were extracted from individual cow sheets of the California Dairy Herd Improvement Association (DHIA) record-keeping system and interfaced with percentage ELISA results for analysis. Univariate and multivariate statistical analyses, using the chi 2 test for categorical variables or Student's t-test for continuous variables and multiple logistic regression respectively, were carried out to evaluate for possible associations between positive antibody tests to each agent and each production variable of interest. Complete data on all variables studied were obtained for 289 (50.5%) cows for M. bovis and 423 (74%) cows for BTV. For cows with complete data on all variables, estimates of the point prevalence of antibodies to BTV and M. bovis were 70.5% and 66.1%, respectively. Results of this study indicated that Guernsey cows were more likely to have a positive BTV test than Holstein cows and that cows in higher lactations were more likely to test positive to BTV ELISA than those in lower lactations (p less than 0.05). Because all cows except those on one farm were Holstein, our confidence in the effect of breed is limited. The association between lactation number and BTV seropositive test may be an age factor identified earlier in the study. For M. bovis, the results of the analysis indicated that seropositive cows were more likely to produce less milk, on a mature equivalent basis (ORadj = 0.96, p = 0.034), and that they had less extended 305 day milk production potential (ORadj = 0.90, p less than 0.0001) than seronegative cows.

A cross-sectional study of bluetongue virus and Mycoplasma bovis infections in dairy cattle: II. The association between a positive antibody response and reproduction performance.

In a cross-sectional study to determine the possible relationship between a positive antibody test to bluetongue virus (BTV) or Mycoplasma bovis infections and reproductive performance of dairy cows, data were collected on 572 California dairy cows during December 1986 for analysis. Serum samples were tested using an enzyme-linked immunosorbent assay (ELISA). Data on reproduction variables were extracted from the individual cow sheets of the California Dairy Herd Improvement Association records and interfaced with the serological results for analysis. Similar data analyses for both BTV and M. bovis were performed to identify and quantitatively assess the association of the reproduction variables and each agent. These associations were evaluated unconditionally using the chi 2 for categorical variables and Student's t-test for continuous variables. Logistic regression analysis was used to determine if reproduction variables with significant unconditional associations remained significant when adjusted for the effects of possible confounding factors. Both the BTV and M. bovis ELISA antibody titres indicated exposure to the agents. The results of the multiple logistic regression analyses indicated that cows seropositive for BTV were significantly older at first calving (p less than 0.03). For M. bovis, seropositive cows were more likely to have longer intervals from calving to last service and longer intervals from calving to pregnancy diagnosis than seronegative cows (p less than 0.05). The other reproduction variables examined were not significantly associated with ELISA seropositivity.

A seroepidemiological study on bluetongue virus in dairy cattle in the central valley of California.

A seroepidemiological study on bluetongue virus (BTV) infection in California dairy cattle was conducted to estimate the prevalence and distribution by age and season of BTV group-reactive antibodies and to look for possible associations between the presence of antibodies and cattle age or breed and farm. Between December 1985 and March 1987, a sample of cattle was tested at approximately two-month intervals for BTV group-reactive antibodies using an enzyme-linked immunosorbent assay (ELISA). Data taken during the month of December 1986 were used to evaluate possible associations between a positive antibody test and certain intrinsic (age, breed) and extrinsic (farm) factors. Univariate and multivariate statistical analyses using the chi-square test for associations and multiple logistic regression, respectively, were carried out for possible associations between positive antibody tests to BTV and each factor of interest. The strengths of the associations were determined using estimates of the odds ratio. Of the 3774 serum samples tested, 238 (6.3%) were from calves, 1045 (27.6%) were from heifers and 2492 (66.0%) were from cows. Seroprevalence varied from nil in calves on two occasions to over 90% on several occasions in cows. Cows consistently had higher prevalence rates than heifers or calves across all test dates (p less than 0.05). The seroprevalence of BTV group-reactive antibodies also showed a seasonal fluctuation, with the highest rates occurring during the warmer months of the year. These highest prevalence rates coincided with heavy activity of the known vector of BTV, Culicoides spp. Breed and farm effects were not statistically significant (p greater than 0.05). With the exception of one farm, all cattle were of the Holstein breed, which reduced confidence in assessing any breed effect in this study. Relative estimates of the sensitivity and specificity of BTV ELISA were 87% and 100% respectively, compared to the standard agar gel immunodiffusion (AGID) test. The observations support previous findings of seasonal distribution of BTV antibodies and suggest an age relationship, whereby older cattle are more likely to be positive to BTV group-reactive antibodies than younger cattle.

The use of the enzyme-linked immunosorbent assay (ELISA) in serological diagnosis of Mycoplasma bovis in dairy cattle.

Between December 1985 and March 1987 an enzyme-linked immunosorbent assay (ELISA) was used with 3774 sera to estimate the prevalence of antibodies to Mycoplasma bovis in sera from three age groups of cattle in four dairies in California and to test for possible associations between the presence of M. bovis antibodies and the age or breed of the cattle and the farm. Unadjusted and adjusted associations were evaluated using the chi-square test for associations and multiple logistic regression analysis, respectively. There was a tendency for the proportion of cattle seropositive for M. bovis to increase steadily and approximately linearly with age (p less than 0.05). There was also a statistically significant relationship between a M. bovis seropositive test and being from Farm IV (p less than 0.05). Farm IV was the largest of the four dairies and this association may be due to the effect of herd size. These findings confirm the ubiquitous distribution of antibodies to M. bovis in dairy cattle in California and also support previous reports of herd size as an important factor in mycoplasmal mastitis.

Serologic and virologic evidence of bluetongue virus infection in cattle and sheep in Mexico.

Three independent 1-year studies were conducted during 3 consecutive years to better define the prevalence of bluetongue virus (BTV) infection in Mexico. Serologic data were obtained by use of agar-gel immunodiffusion for identification of BTV group-reactive antibodies, and virologic data were obtained by virus isolation. Samples were obtained from sheep in 6 states over a 1-year period, with 9% seropositive; samples were obtained from cattle in 11 states during the same 1-year period, with 35% seropositive. Two years later, samples were obtained from cattle in 4 additional states, with 69% seropositive. Virus isolation was conducted on pooled blood samples obtained from cattle in 7 states. Six virus isolates were recovered and included 2 isolates each of BTV serotypes 11 and 13 and 1 isolate each of serotypes 10 and 17. All virus isolates were partially characterized by electrophoretic analysis of genomic RNA migration profiles (electropherotypes) in polyacrylamide gels. All Mexican isolates of BTV differed considerably in electropherotype profile, as compared with their respective US prototype strain of the same serotype. Such differences appeared to be much more extensive than those described to exist between numerous California isolates of the same serotype.

IgG, IgM and IgA in the serum of cattle naturally infected with Mycobacterium paratuberculosis.

Serum IgG, IgM and IgA antibody response in 20 cattle naturally infected with Mycobacterium paratuberculosis and in 15 non-infected cattle were measured by enzyme-linked immunosorbent assay. A strong IgG response was detected in 16 (80%) of the infected animals. Diagnostic levels of IgM were detectable in all of the infected animals as well as in 8 (53%) of the non-infected animals. Animals with paratuberculosis had a very weak specific serum IgA response and this appears to be of little value in detection of infection in these animals.

Evaluation of the Minitek system for characterization of Bacillus species.

Minitek (BBL Microbiology Systems, Cockeysville, Md.) substrate disks were evaluated as alternatives to conventional tests for the characterization of Bacillus species. Results were compared for 10 reference isolates and 87 isolates from food sources. The overall agreement of results between the Minitek and conventional tests was 92% for reference strains and 86% for food isolates.