Antibodies to bovine leukemia virus and presence of malignant lymphoma in slaughtered California dairy cattle.

Rates of malignant lymphoma (ML) in California dairy cattle and relationships between ML and presence of antibodies to bovine leukemia virus (BLV) were investigated. An excessively high rate of lymphoma was found in the population studied and a previously unknown relationship between ML and p-24 antibodies to BLV was identified. Sera from slaughtered California holstein dairy cows were tested by agar gel immunodiffusion for presence of antibodies to the gp-51 and p-24 antigens of BLV. Of the 7,760 cows slaughtered, 32% and 10% had gp-51 and p-24 antibodies, respectively, and 52 cows (0.67%) were condemned for ML. Lymphoma was diagnosed histopathologically in 37 (86%) of 43 cows sampled from those 52 condemned. Highly elevated prevalence rates of ML in gp-51 and p-24 antibody-positive cows were 172.6 and 511.5 per 10,000 cows, respectively. A prevalence rate of lymphoma not associated with presence of BLV antibodies was estimated for the first time to be 1.55 per 10,000 cows. A highly significant (P less than .00001) and strong (odds ratio = 26.0) association was found between presence of p-24 antibodies and presence of ML for cows with gp-51 antibodies. When results of serologic tests were compared in regard to diagnosis of ML, the highest sensitivity was with use of gp-51 serology alone (97.3%) and highest specificity was with use of p-24 serology alone (90.4%). False-negative and false-positive rates of diagnosis of ML for gp-51-positive, p-24-negative and gp-51-positive, p-24-positive cows were 0.21 and 94.8%, respectively.

Immunohistological demonstration of virus and tumor associated antigens in tissues experimental and spontaneous bovine leukemia virus (BLV) infection.

Expression of bovine leukemia virus (BLV) antigens in vivo has not been shown. After BLV infection, however, production of antibodies directed towards BLV proteins (e.g. gp51) can be easily demonstrated. Thus, production of BLV proteins has to take place somewhere in infected cattle. Tissues and organs of experimentally infected cattle were fixed in acetone and embedded in paraffin. Monoclonal antibodies directed to gp51 were used to demonstrate BLV expression immunohistologically by the peroxidase-antiperoxidase (PAP) method. The same samples were also used to demonstrate a tumor associated antigen (TAA) employing a monoclonal antibody. Our results indicate that very few cells, found in the intestinal mucosa, produce gp51 in vivo. The expression of TAA, however, increases significantly shortly after infection with BLV and remains high throughout life.

Alteration in lymphocyte subpopulations in bovine leukosis virus-infected cattle.

Alterations in peripheral blood lymphocyte subpopulations were examined in bovine leukosis virus (BLV)-infected cattle using antibodies specific for differentiation antigens in conjunction with analytical flow cytometry. Animals considered to be aleukemic and lymphocytotic were included in the study. Significantly fewer numbers of circulating B-lymphocytes (surface Ig-positive) and T-helper lymphocytes (BoCD4-positive) were identified in BLV-infected aleukemic cattle compared to non-infected controls while no significant differences were established for T-cytotoxic/suppressor lymphocytes (BoCD8-positive). In contrast, BLV-infected animals with persistent lymphocytosis had elevated numbers of circulating B-lymphocytes with no significant perturbation in circulating T-lymphocyte subsets identified when compared as a group with the negative control cattle. Application of regression analysis to data from individual lymphocytotic cattle demonstrated a significant correlation between absolute numbers of B- and T-lymphocytes. Increased numbers of B-lymphocytes were correlated with increased numbers of T-helper and T-cytotoxic/suppressor lymphocytes.

Prospective serology and analysis in diagnosis of dairy cow abortion.

A prospective serologic investigation was undertaken on 3 California dairies (herds 1, 3, and 4, as previously designated in a report on abortion surveillance) to determine if fetal loss was associated with infectious disease agents in cows. The diagnostic problem in these herds was typical of many dairies in that abortions were not discovered for several months and aborted fetuses were seldom recovered. Blood from approximately 100 pregnant cows in each herd was sampled at monthly intervals, beginning when the cows were palpated at approximately 40 days gestation. Sera were tested for antibodies to infectious bovine rhinotracheitis virus and bovine viral diarrhea virus (BVDV) and to the Leptospira serovars pomona, hardjo, grippotyphosa, icterohemorrhagiae, canicola, and szwajizak. Logn antibody titers were examined for an association with fetal loss, using multivariate methods of logistic regression and survival analysis. Of the 325 cows followed, 37 aborted, and no fetuses were recovered. Statistical analyses indicated that significant fetal loss was associated with high titers to L. hardjo and with low titers to L. szwajizak (herds 1 and 4) and BVDV (herd 1). Results for herd 3 revealed a connection between abortion and L. icterohemorrhagiae (P = 0.036) and L. canicola (P = 0.050) and possible vaccinal protection against abortion caused by L. grippotyphosa (P = 0.027 and 0.015). For herd 4, there was a marginally significant tendency for the first vaccination of the gestation against leptospirosis to have protected against fetal death (P = 0.077). Advantages of the diagnostic design were that it permitted comparison of titers of aborted cows with those of nonaborted cows and it considered vaccination-induced titers in analyses.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of birthweight, total protein, serum IgG and packed cell volume on risk of neonatal diarrhea in calves on two California dairies.

The objective of the study was to determine if there was a relationship between hematological, immunological and physiological variables of newborn calves and risk of diarrhea during the neonatal period. Four hundred and seventeen heifer calves from two dairies (A and B) in the San Joaquin Valley of California were enrolled at birth and scored daily, to 28 days of age, for evidence and severity of diarrhea (0 to 3). Calves were weighted at birth and blood sampled at two to five days of age to determine packed cell volume (PCV), total protein (TP) and IgG serum concentration. The Cox proportional hazards model was used to determine if age at onset of the first diarrhea episode and length of the first episode were associated with the hypothesized variables (PCV, TP, IgG and birthweight). The IgG concentration was not associated with the age at onset of diarrhea (p = 0.6052, Dairy A; p = 0.4393, Dairy B) but a high IgG concentration was associated with a decreased length of episode (p = 0.0325, Dairy A; p = 0.0912, Dairy B), particularly for calves born in the winter on dairy A (p = 0.0211). For calves born in the winter, those with either a high or a low birthweight had diarrhea at a younger age (p = 0.0102, Dairy A; p = 0.0020, Dairy B). Associations were also found for PCV and TP with both the age at onset and length of the first episode of diarrhea. Results suggest that parameters measurable at, or shortly after birth may have important prognostic value in evaluating risk of calf diarrhea.

Effect of brucellosis vaccination and dehorning on transmission of bovine leukemia virus in heifers on a California dairy.

Brucellosis vaccination and dehorning were examined for an association with bovine leukemia virus (BLV) infection in heifers on a California dairy between April 1984 and June 1987. Between December 1985 and June 1986, weaned heifers were dehorned using the gouge method at the time of brucellosis vaccination. Using logistic regression, the estimated probability for a nondehorned heifer to seroconvert within three months after brucellosis vaccination (0.08) was significantly less than that for heifers dehorned after a noninfected heifer (0.46) or than that for heifers dehorned after an infected heifer (0.85) (p = 0.039 and p less than 0.001, respectively). To evaluate risk of transmission by brucellosis vaccination, which was usually done within one month postweaning, cumulative proportions of heifers remaining uninfected were computed among heifers that did not seroconvert three months after dehorning. Because results of a Cox model analysis indicated that groups of heifers were 6.6 times more at risk of becoming infected if placed in pens holding gouge-dehorned heifers (where prevalence varied between 50 and 70%) (p less than 0.001) than other groups placed in pens without gouge-dehorned heifers (where prevalence varied between 10 and 30%), cumulative proportions of heifers remaining uninfected were computed for each type of group. The cumulative proportion of heifers remaining uninfected from weaning to first calving was 0.60 for the high prevalence group and 0.96 for the low prevalence group. No change in slope of cumulative proportions was observed before and after one month postweaning, suggesting that brucellosis vaccination was not an effective means of transmission.(ABSTRACT TRUNCATED AT 250 WORDS)

Upper-normal prediction limits of lymphocyte counts for cattle not infected with bovine leukemia virus.

A study was conducted to develop valid estimates of lymphocyte count (LC; cells per microliter) of individual, clinically normal dairy cattle. Estimated weighted regression was used on repeated measures of individual LC to examine 6 models predicting LC as a function of age in cattle not infected with bovine leukemia virus. The generalized growth curve model of analysis of variance was used to estimate intercepts, slopes, and prediction limits for the models and to compare the LC-to-age relationship between Holstein and Guernsey breeds. The best-fitting model (P = 0.0001) with the narrowest prediction interval was LC = 4,414.4 - 84.6X, where X = (age -48) if age less than or equal to 48 months, and X = 0 if age greater than 48 months, and 163.6 and 8.1 are the SE of the estimates, respectively. Upper one-sided 95%-predicted normal LC tended to be higher than estimates derived from traditional hematologic keys that use confidence limits of mean LC. Difference was not found in the LC-to-age relationship between the Holstein and Guernsey cattle (P = 0.67). Results of this study provided estimates of normal LC that are more specific in diagnosing lymphocytosis in individual cattle.

Effect of vaccination with the R mutant Escherichia coli (J5) antigen on morbidity and mortality of dairy calves.

A prospective cohort study was undertaken to determine the efficacy of Escherichia coli 0111:B4 (J5) vaccination in dairy calves. Calves on 2 units were vaccinated when they were 2 to 3 days old and 2 weeks later with the J5 antigen or they were left unvaccinated, and were observed during the first 60 days of age for morbidity and mortality. Events recorded were death, the first case of illness, the first sign of a respiratory tract condition, the first sign of diarrhea, and the first treatment. The time to death or to a morbid event was examined as a function of vaccination status, using the Cox model of survival analysis, where serum IgG concentration at 2 to 3 days of age and gender of the calf were included to control confounding. Signs of morbidity in 517 calves were followed, 189 from unit 1 and 328 from unit 2. Vaccination was associated with a 2.15-fold reduction in risk of death on unit 2 (P = 0.042), but with a 2.43-fold increase in risk of death on unit 1 (P = 0.0035). The only association found between vaccination and morbidity was a 1.34-fold reduction in risk of respiratory tract signs for vaccinated calves on unit 2 (P = 0.055). Necropsy results and clinical investigations indicated that calves on unit 1 were poorly nourished and emaciated. Antibodies to J5 2 weeks after vaccination were significantly (P = 0.0002) lower in calves on unit 1 than in calves on unit 2. The findings offered a possible explanation for the adverse vaccination effect in calves on unit 1.(ABSTRACT TRUNCATED AT 250 WORDS)

Fetal loss associated with palpation per rectum to diagnose pregnancy in cows.

A prospective observational study was performed to determine whether palpation per rectum of cows in the first 6 weeks of gestation to diagnose pregnancy contributed to fetal attrition. Pregnancy diagnoses were made by private practitioners as part of their routine herd-health service on 9 dairies in the San Joaquin Valley of California. To determine whether there was an association between abortion and fetal age at time of palpation, the probability of abortion was tested as a function of fetal age at palpation, controlling for possible modifying and confounding effects of herd, age at conception, gravidity, parity, and number of days-in-lactation at conception. Results of logistic regression analyses for 19,411 pregnancies followed for up to 90 days after palpation indicated that, during the 28- to 42-day period, palpation of fetuses earlier in the period was associated with a significantly (P < 0.0001) low probability of abortion, compared with that for palpation later in the period. An association between abortion and palpation of fetuses > 42 days of age was not found. Results were suggestive that, given conditions and techniques typical of private practice, fetal death may not be a usual manifestation of early palpation of cows to diagnose pregnancy, rather, that there may be a slight increase in risk of fetal death as the fetal age at palpation increases from 28 to 42 days.

Effect of intrauterine antimicrobial treatment in reducing calving-to-conception interval in cows with endometritis.

A prospective clinical trial was conducted on 2 large dairies in the San Joaquin Valley of California to determine whether a single intrauterine infusion with procaine penicillin G or oxytetracycline reduced the calving-to-conception interval in cows with endometritis. Cows with endometritis were randomly assigned to a treatment or a control group. The uterus of treated cows on 1 dairy was infused with 0.8 to 1.0 million U of procaine penicillin G in 40 ml of sterile water, and the uterus of treated cows on the other dairy was infused with 500 mg of oxytetracycline in 20 ml of sterile water, both of which were typical doses used on dairies in the area. A difference was not observed in the cumulative proportion of cows remaining nonpregnant between 87 penicillin-treated and 77 control cows on the 1 dairy (P = 0.356), or between 74 oxytetracycline-treated and 62 control cows on the other dairy (P = 0.174). Results suggest that routine infusion of antibiotics to treat endometritis, as commonly practiced, may not be efficacious.

Considerations for use of descriptive epidemiology to investigate fetal loss in dairy cows.

Various descriptive approaches were taken in a prospective investigation to characterize fetal loss in cows on a California dairy. The methods and observations were offered for consideration by practitioners engaged in dairy herd health medicine. For 4,732 pregnancies followed from 2,163 cows in a 6.5 year period, the respective proportions (percentage) of cows aborting (1--cumulative proportion not aborting by 260 days) and abortion densities (abortions per 10,000 cow-days-at-risk) were 10.63 and 6.29 for all fetal deaths, 9.36 and 5.49 for deaths resulting in fetal expulsion, and 1.39 and 0.80 for deaths resulting in mummification. The greatest risk of fetal death (119 deaths/10,000 fetuses/d) was observed between 98 and 105 days of gestation, and median age at fetal death ranged from 99.0 to 105.5 days. Abortion density for fetal deaths resulting in mummification for cows conceiving during September (1.61/10,000 fetuses/d) and October (1.63/10,000 fetuses/d) was tenfold greater than that for cows conceiving in February (0.16/10,000 fetuses/d) and was twice that of the overall rate (0.84/10,000 fetuses/d). For cohorts of nonculled cows, abortion rate increased after 5 years of age, after 5 pregnancies, or after 4 calvings. For cows with at least one previous abortion, the proportion aborting (14.50%) was higher than that for cows without a previous abortion (12.14%). For a given gravidity, abortion rate was higher among cows that had experienced a previous abortion, compared with those that had not. These methods and observations may help provide a logical foundation on which to base clinical hypotheses regarding causes of abortion, and they may offer insight into pitfalls of bias and confounding to be anticipated.

Vesicular stomatitis virus (New Jersey strain) infection in two California dairy herds: an epidemiologic study.

The course of vesicular stomatitis in cattle was investigated in 2 dairy herds (A and B) located in the southern San Joaquin Valley of California. Cattle were examined and specimens were obtained for virus isolation and for serologic survey for one year after an epizootic in December 1982. All 33 lactating cows selected for study had oral lesions, but only 19 (58%) were drooling or frothing around the mouth. Lesions on feet and teats were not observed. The healing time (longer than has been reported previously) for oral lesions ranged from 34 to 59 days. The mean serum neutralizing antibody titer for all cows tested in both herds 21 days after clinical signs were first observed was greater than 1:512. The mean titer decreased in the first 11 months after the epizootic, but remained greater than 1:128, and then increased during December 1983. Vesicular stomatitis virus/New Jersey strain was not isolated from 239 blood samples, 235 swab specimens of oral cavities, 38 swab specimens of oral epithelium, 206 urine specimens, or 232 fecal specimens collected from cows; however, it was isolated from tongue epithelium of 3 cows at 1, 4, and 21 days after signs of frothing were first noticed. For 20 lactating cows brought into dairy A during the epizootic, a mean time of 8.9 days elapsed between time of entry and appearance of clinical signs of vesicular stomatitis.(ABSTRACT TRUNCATED AT 250 WORDS)

Prevalence of salmonellae in cattle and in the environment on California dairies.

Milk samples were collected from all lactating cows on 60 dairies (mean number of cows/dairy, 584; range, 66 to 2,834) randomly selected from 701 California dairies enrolled in the Dairy Herd Improvement Association program. Samples were tested, by means of an ELISA, for antibodies against Salmonella serogroup B, C1, and D1 antigens (somatic antigens O1, 4, 6, 7, 9, 12). Blood samples were collected from all cows with positive results and tested for serologic evidence of exposure to salmonellae. Samples for bacteriologic culture (pooled feces from 20 randomly selected calves, swabs of wet areas and feces from calf pens and dairy hospital pens, drag swab sample from wastewater lagoon, and samples of feed components) were also collected from all 60 dairies. Seven (11.7%) of the 60 dairies each had 1 sample that yielded Salmonella organisms (3 S typhimurium, 1 S dublin, 1 nonmotile Group D salmonella, 1 S derby, and 1 S oranienberg). Five of the Salmonella isolates came from the hospital pens and 2 came from calf pens. Thirty-three dairies did not vaccinate cattle against salmonellosis, and of these, 24 (72.7%) had > or = 1 seropositive cow (titer > or = 200), and 20 (61%) had > or = 1 persistently seropositive cow (titer for each of 2 blood samples collected > or = 60 days apart was > or = 200). Of the 27 dairies that did vaccinate cows against salmonellosis, 24 (89%) had > or = 1 seropositive cow, and 21 (78%) had > or = 1 persistently seropositive cow.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of abortions in cattle attributable to neosporosis in selected dairy herds in California.

OBJECTIVE: To estimate the minimum rate of abortion attributable to infection with Neospora sp in selected California dairy herds. DESIGN: Prospective study. ANIMALS: Twenty-six dairy herds containing 19,708 cows were studied. Fourteen herds had a history of abortions attributable to neosporosis, and 12 were herds in which neosporosis had not been identified as a cause of abortions. PROCEDURE: During a 1-year period, all available aborted fetuses were submitted to veterinary diagnostic laboratories to determine the cause of abortion. Reproductive records of cows that aborted were reviewed. RESULTS: Neospora sp infection was the major cause of abortion identified (113/266 abortions, 42.5%). The majority (232/266, 87.2%) of the aborted fetuses were submitted from herds with a history of abortions attributable to neosporosis, and Neospora sp infection was identified as the causative agent in 101 of 232 (43.5%) of the abortions from these herds. Fewer aborted fetuses were submitted from the 12 herds that did not have a history of abortion attributable to Neospora sp; however, neosporosis was confirmed as a cause of abortion in 6 of these 12 herds and was identified as the causative agent in 12 of 34 (35.3%) abortions from these herds. The disease was widespread throughout the state (19/26 herds in our study). Available reproductive histories of cows that had abortions attributed to neosporosis were evaluated, and 4 cows were identified that twice aborted Neospora-infected fetuses. CLINICAL IMPLICATIONS: Abortion attributable to Neospora sp infections can be expected to be a continuing major cause of abortion in dairy herds with a history of neosporosis as well as in dairy herds that have a history of sporadic abortions, but for which Neospora sp infections have not been previously identified as a cause of abortion. Subsequent pregnancies in cows that abort a Neospora sp-infected fetus also are at risk of infection, suggesting that the immunity provided by an initial infection is inadequate to prevent repeat infection or that cows can be persistently infected with Neospora sp.

Development of computational models for the purpose of conducting individual livestock and premises traceback investigations utilizing National Animal Identification System-compliant data.

Many of the efforts surrounding the development of the National Animal Identification System have encompassed the identification of livestock production and handling premises as well as individuals or herds of animals, whereas little effort has been directed toward the ultimate goal of animal traceback within 48 h. A mock data set representative of the Colorado cattle population was created for modeling of cattle traceability. Using this data set, algorithms were developed to complete rapid and accurate traceback and traceforward of animals or premises or both. On July 19, 2005, the Colorado Department of Public Health and Environment, in conjunction with the Colorado Department of Agriculture, conducted a test exercise pertaining to homeland security. The exercise team randomly identified animal number 926,583 (of the 2 million total animals) as a potentially infected animal of interest and requested a traceback of this animal. Traceback was accomplished in 215 s, and 540 primary coresident animals were identified. However, due to animal movements, the number of coresidents (animals exposed, directly or indirectly, to the animal of interest) expanded with coresidency level (level 1 = direct contact; level 2 = direct contact with an animal that had direct contact with the animal of interest; level 3 = direct contact with an animal that had contact with an animal that had direct contact with the animal of interest, etc.) to more than 1.2 million coresidents at level 4, and more than 90% of all animals identified as a coresident at some level. In addition to the coresidency results, the premises containing the coresidents were identified and sorted by the number of coresidents. Because of animal movement, all 19,391 premises included in the data set had coresidents at some level. This exercise demonstrated the capability of the developed algorithms to complete rapid traceback and the complexity of the resulting animal traceback output.

Factors associated with transmission of bovine leukemia virus by contact in cows on a California dairy.

A prospective study was undertaken on a California dairy from 1984 to 1987 to examine factors associated with contact transmission of bovine leukemia virus in cows. Two approaches were used to model the probability of infection. First, the expected number of new infections per pen-month was assumed to follow a binomial distribution. The probability of infection was modeled, using logistic regression, as a function of prevalence of infection in pen, presence of lactating cows, proportion of pregnant cows, presence of an infected bull, and proportion of infected cows with a lymphocyte count above 10,000 cells/microliters of blood and/or with Mr 24,000 protein (p24) antibodies. The probability of infection was significantly associated with the prevalence of infection and at the limit of significance for presence of lactating cows in pen. Second, the Cox model with time-dependent covariates was used to analyze time from first parturition or from the beginning of the study to infection. Factors examined were age, breed, lactation number, whether the cow was pregnant and/or lactating, prevalence of infection, presence of an infected bull, and density of cattle in the pen holding the cow. Non-pregnant cows were 2.9 times more at risk of infection than were pregnant cows, and risk of infection significantly increased as prevalence of infection in a pen increased.

Serum zinc and copper concentrations in relation to spontaneous abortion in cows: implications for human fetal loss.

The purpose of this study was to investigate the effect of the status of dietary zinc and serum zinc and copper concentrations on the risk of fetal loss in 570 cows. three herds received no supplements (herds 1, 3, 4), while cows in herd 2 received supplements of either 7 g zinc week-1 (n = 118), as zinc methionine, or a control diet containing methionine (n = 128). Serum zinc, copper and metallothionein concentrations were determined once a month throughout gestation. Logistic regression and survival analysis were used to examine for associations between risk of fetal loss and serum zinc, copper, copper:zinc, or metallothionein concentrations, supplement level, and maternal age at conception. The risk of fetal loss increased when both serum zinc decreased and copper concentrations increased (P < 0.0001; relative risk = 10.28, 95% confidence intervals = 4.69, 22.5). The attributable risk, for a decline in the zinc concentration by 10 mumol l-1 and an increase in the copper concentration by 5 mumol l-1 was 90.27%. Methionine-supplemented cows had a higher risk of fetal loss compared with zinc-methionine-supplemented cows (one-tailed P = 0.0375; relative risk = 2.98). Cows in herds 1, 3 and 4 had a higher risk for abortion than did zinc-methionine-supplemented cows in herd 2 (relative risk = 26.27, 95% confidence intervals = 2.31, 299.38; relative risk = 40.87, 95% confidence intervals = 3.50, 458.43; relative risk = 41.53, 95% confidence intervals = 3.77, 457.02, respectively). Our results suggest that inflammation and zinc nutriture may play an important role in fetal loss in dairy cows.

The effect of pre-enrichment on recovery of Streptococcus agalactiae, Staphylococcus aureus and mycoplasma from bovine milk.

The study was conducted to determine whether pre-enrichment would increase sensitivity of detecting Streptococcus (Str.) agalactiae, Staphylococcus (S.) aureus, and mycoplasma in bovine milk. Two procedures were followed, one involving direct inoculation of milk on bovine blood agar, and the other involving preenrichment in broth followed by inoculation on agar. Logistic regression was used to predict the probability of isolation as a function of culture procedure and two additional covariates, the California Mastitis Test (CMT) score of the milk and the type of sample (indicating sample storage temperature and herd mastitis status). A total of 13778 milk samples was cultured for each of the three bacteria. By using results of both direct inoculation and pre-enrichment, the probability of isolation compared to use of direct inoculation only and adjusted for effects of other variables was increased 3.6-fold for Str. agalactiae, 1.6-fold for S. aureus and 1.7-fold for mycoplasma. The probability of isolation for all three bacteria increased as the CMT score increased. For Str. agalactiae, there was a statistical interaction predicting that enrichment improved the odds of isolation more from milk with high CMT scores than from milk with low scores. Results indicate that pre-enrichment can substantially increase the sensitivity of bacteriological screening of dairy cows for mastitis caused by Str. agalactiae, S. aureus, and mycoplasma.