Detection of asymptomatic renal Leptospira infection in abattoir slaughtered cattle in southeastern Georgia, United States.

OBJECTIVES: Leptospirosis is one of the most widespread zoonotic infectious diseases affecting humans and animals. Several animal species, including cattle, can act as potential asymptomatic carriers facilitating zoonotic transmission of Leptospira. This study was conducted to assess the occurrence of asymptomatic renal Leptospira carriers among cattle slaughtered in southeastern Georgia, United States. METHODS: A battery of diagnostic tests, including dark field microscopy, direct fluorescent antibody staining, polymerase chain reaction, and culture, were performed on a set of bovine kidneys (n = 37) collected from an abattoir in southeastern Georgia, United States. Virulence of a field isolate obtained from this study was tested in a hamster experimental model. RESULTS: Motile spirochete-like structures were observed by dark field microscopy in 23 (59%) out of 37 kidney samples tested. In all, 29 samples (78%) were positive by direct fluorescent antibody staining. Only 11 (29.7%) samples by polymerase chain reaction and 3 (8.1%) by culture were positive for Leptospira sp. The isolates obtained by culture were confirmed as Leptospira borgpetersenii. Hamsters experimentally infected with one of the Leptospira field isolates obtained from this study did not show clinical signs but developed renal infection with interstitial nephritis and tubular necrosis. CONCLUSIONS: This study confirms that asymptomatic Leptospira renal infection is present among cattle in the region. Our findings underscore the need for future studies to assess the potential environmental contamination and transmission to humans in contact with infected cattle.

Effect of calf age and administration route of initial multivalent modified-live virus vaccine on humoral and cell-mediated immune responses following subsequent administration of a booster vaccination at weaning in beef calves.

OBJECTIVE: To compare immune responses following modified-live virus (MLV) vaccination at weaning after intranasal or SC administration of an MLV vaccine to beef calves at 2 or 70 days of age. ANIMALS: 184 calves. PROCEDURES: Calves were allocated to 1 of 5 groups. The IN2 (n = 37) and IN70 (37) groups received an MLV vaccine containing bovine herpesvirus 1 (BHV1), bovine viral diarrhea virus (BVDV) types 1 and 2, bovine respiratory syncytial virus (BRSV), and parainfluenza 3 virus intranasally and a Mannheimia haemolytica and Pasteurella multocida bacterin SC at median ages of 2 and 70 days, respectively. The SC2 (n = 36) and SC70 (37) groups received a 7-way MLV vaccine containing BHV1, BVDV1, BVDV2, BRSV, parainfluenza 3 virus, M haemolytica, and P multocida SC at median ages of 2 and 70 days, respectively; the control group (37) remained unvaccinated until weaning. All calves received the 7-way MLV vaccine SC at median ages of 217 (weaning) and 231 days. Serum neutralizing antibody (SNA) titers against BHV1, BVDV1, and BRSV and intranasal IgA concentrations were determined at median ages of 2, 70, 140, 217, and 262 days. Cell-mediated immunity (CMI) against BHV1, BRSV, BVDV1, and P multocida was determined for 16 calves/group. RESULTS: At median ages of 140 and 217 days, BVDV1 SNA titers were significantly higher for the SC70 group than those for the other groups. Intranasal IgA concentrations and CMI increased over time for all groups. Vaccination at weaning increased SNA titers and CMI in all groups. CONCLUSIONS AND CLINICAL RELEVANCE: SC administration of an MLV vaccine to 70-day-old calves significantly increased BVDV1 antibody titers before weaning.

Humoral immunity and injection-site reactions in cattle vaccinated with a multivalent clostridial vaccine administered via subcutaneous injection or via transdermal needle-free injection.

OBJECTIVE: To evaluate injection-site reactions and serum antibody titers in cattle vaccinated with a clostridial vaccine administered SC or via needle-free transdermal injection. ANIMALS: Sixteen 11-to 12-month-old Herefords. PROCEDURES: Cattle in 2 groups were vaccinated on days 0 and 28 with a commercially available multivalent clostridial vaccine administered SC or transdermally Injection sites and serum antibody titers were evaluated at several time points after vaccination. Serum antibody titers against Clostridium perfringens beta toxin, Clostridium novyi alpha toxin, and Clostridium septicum alpha toxin were determined with an ELISA; Clostridium sordellii lethal toxin titers were determined with a toxin neutralization assay. RESULTS: Firm injection site swellings developed in cattle vaccinated via either route; however, at several observation times, swellings were significantly smaller in cattle vaccinated transdermally. Serum titers against C perfringens beta toxin and C septicum alpha toxin did not differ significantly between groups after vaccination; serum titers against C novyi alpha toxin were not significantly different between groups, except on days 10 and 56, when they were significantly higher in cattle vaccinated SC. Titers against C sordellii lethal toxin were significantly higher in cattle vaccinated SC on several days after vaccination, but titers were not significantly different after day 49. CONCLUSIONS AND CLINICAL RELEVANCE: Transdermal vaccination of cattle resulted in serum antibody titers that were similar to those induced via SC vaccination and caused injection-site reactions that were significantly smaller. Transdermal vaccination may be an effective technique for vaccinating cattle against clostridial diseases while minimizing local reactions that often develop after clostridial vaccination.

Comparison of fluorescent antibody and microscopic agglutination testing for Leptospira in pregnant and nonpregnant cows.

Serum and urine samples from 30 cows (15 pregnant and 15 nonpregnant) from each of 10 Georgia dairy herds (total cows = 300) were examined by microscopic agglutination testing (MAT) and direct fluorescent antibody testing (FAT), respectively. Seven of the 10 herds had at least 1 cow with a positive FAT, and all of the herds had at least 1 cow with a reciprocal MAT titer > or =100 for 1 or more serovars. Serological testing was not helpful in identifying the infecting serovar for cows with a positive FAT result. The MAT titers for all 7 of the serovars evaluated were significantly correlated with one another, with 17 (81%) of the 21 Spearman rank correlation coefficients > or =0.4 in magnitude. Twenty (56%) of 36 FAT-positive cows did not have a titer that was highest for any particular serovar. Four of the 7 herds that reported using a Leptospira borgpetersenii serovar Hardjo-bovis vaccine had one or more FAT-positive cows compared with 3 out of 3 herds that reported they were not using this type of vaccine, although this difference was not statistically significant.

Brain heart infusion broth may not be a required component for the decontamination process for the isolation of Mycobacterium avium subspecies Paratuberculosis from fecal samples using ESP broth cultures.

Based on the authors' laboratory experience indicating that increased bacterial contamination in Mycobacterium avium ssp. paratuberculosis (MAP) cultures may be because of the addition of brain heart infusion broth (BHI) during the decontamination process, this study was designed to examine whether BHI is a required component for the isolation of MAP from ESP(R) broth cultures. Twenty-six National Veterinary Services Laboratory (NVSL) proficiency test samples supplied for the year 2005 were used for the comparison. Two paired sets of samples were processed in the experiment. In one set, the hexadecylpyridinium chloride monohydrate (HPC) and antibiotic brew were prepared in half strength BHI and for the other set, HPC and antibiotic brew were prepared in sterile distilled water. Culture of the 26 samples using the BHI/HPC decontamination method identified 13 (50%) positives, whereas culture using the water/HPC decontamination method identified 14 (54%) positives. The proportions of samples with a positive result did not differ significantly between the 2 decontamination methods. Although in most cases it took longer to identify a positive result by the BHI method, the difference between methods with respect to the number of days to a positive culture result was not statistically significant. Retrospective data collected from the Animal Disease Diagnostic Laboratory, Ohio also suggest that inclusion of BHI in the decontamination protocol may not have any effect on MAP recovery or contamination rate. Elimination of BHI from broth cultures may increase the sensitivity of MAP isolation, and reduce the cost of testing.

Prevalence of Mycoplasma bovis in backgrounding and stocker cattle operations.

OBJECTIVE: To characterize the prevalence of Mycoplasma bovis infection in backgrounding and stocker cattle operations and compare bacteriologic culture with PCR assay for detection of M bovis. DESIGN: Prospective descriptive study. ANIMALS: 432 calves, 3 to 9 months old, from 9 operations. PROCEDURES: 2 nasal swab specimens were collected from each calf. Swab specimens were evaluated via bacteriologic culture and PCR assay for organisms of the class Mollicutes and M bovis. Culture results were considered negative if no growth occurred within 21 days. Positive results were indicated by characteristic colony formation with PCR assay confirmation. Deoxyribonucleic acid was extracted from 1 swab specimen for direct PCR assay for Mollicutes and M bovis. RESULTS: Of 432 calves, 374 (87%) had positive results for Mollicutes via PCR assay and 63 (15%) via culture. Seven (2%) calves had positive results for M bovis via PCR assay and 10 (2%) via culture. Prevalence of Mollicutes at the farm level ranged from 54% to 100% via PCR assay and from 0% to 59% via culture. Prevalence of M bovis at the farm level ranged from 0% to 4% via PCR assay and from 0% to 6% via culture. Calves that shed M bovis were significantly more likely to have a fever than were calves that did not shed M bovis. CONCLUSIONS AND CLINICAL RELEVANCE: M bovis was detected at a low level in recently purchased backgrounded and stocker calves in Georgia. Although slightly more infected calves were detected via culture and PCR assay together, PCR assay appeared to accurately identify M bovis at the farm level.

Efficacy of spheroplastic and cell-wall competent vaccines for Mycobacterium avium subsp. paratuberculosis in experimentally-challenged baby goats.

A Mycobacterium avium subspecies paratuberculosis (MAP) vaccine that reduced the incidence of clinical disease or reduced fecal shedding of MAP would aid control of Johne's disease (JD). The objective of the present study was to evaluate the efficacy of four MAP vaccine combinations, including cell-wall competent (CWC) alum adjuvant, CWC-QS21 adjuvant, cell-wall deficient (CWD) alum adjuvant and CWD-QS21 adjuvant vaccines. Eighty baby goats were vaccinated at 1 and 4 weeks of age with one of these vaccines or a sham control vaccine consisting of alum adjuvant. Kids were challenged orally with approximately 6.0x10(9) organisms in four divided doses of 1.5x10(9) organisms using a goat isolate of MAP. Vaccinated challenged and challenged control groups had 10 and 6 kids per group, respectively. Half of the kids within each group were necropsied at either 6 or 9 months post-challenge. Gross and microscopic lesions and relative number of acid-fast bacilli were evaluated and scored at necropsy. Results indicated all challenged kids had some lesions compatible with JD suggesting none of the vaccines prevented infection. Three vaccines (CWC-alum, CWC-QS21 and CWD-QS21) reduced lesion scores by 46-51% at 9 months. CWD-alum vaccine resulted in a more severe (+33.5%) lesion score than sham-vaccinated challenged control. Lesion scores were greater at 9 months than at 6 months post-challenge in the sham-vaccinated challenged group and CWD-alum vaccinated group, while lesion scores were generally stable with remaining vaccines. Mean fecal CFU/g were significantly different across time from challenge to 9-month necropsy (p=0.043) and the CWC-QS21 vaccine group had a marked reduction in fecal CFU/g at all time points post-challenge. A reduction in MAP CFU/g was also detected in necropsy tissues from kids given the CWC-alum, CWC-QS21 and CWD-QS21 vaccines, and increased CFU/g were detected in tissues from kids given the CWD-alum vaccine. Immunological tests evaluated included, humoral response evaluation by AGID, ELISA and Western blot, and cell mediated response by comparative PPD skin testing (M. avium, Old Johnin, M. bovis and Lot 2 Johnin PPD's), and production of MAP induced gamma-interferon. Vaccination also resulted in false-positive PPD skin test reactions for M. avium PPD, Old Johnin PPD and gamma-interferon tests. When a 2-mm cutoff above normal skin thickness was used to define positive skin test reactions, false-positive reactions for M. bovis were detected in only 2 of 32 kids given a vaccine with QS21 adjuvant.

Teaching veterinary students beef production medicine with student/producer teams.

Veterinary colleges face difficulties in meeting the demand for rural veterinarians with the scope to practice quality production medicine. Increasing population density around veterinary colleges, retaining the interest of students with a background in animal agriculture, and educating students without a farm background requires that veterinary colleges consider innovative ways to not only teach traditional food-animal practice but give future veterinarians the advanced skills the food industry demands. This article describes a three-year elective program, Beef Records Analysis, in which beef production medicine is taught by teaming a student and a beef producer together early in the student's veterinary education. These producer/student teams complete risk assessments, balance rations, collect financial and production information, and evaluate back-grounding and feedlot enterprises. Students learn how to evaluate their producers using industry benchmarks and past performance records and how to communicate their findings back to their producer. Producers often make management decisions based on the students' findings, and, because the students maintain their relationships with producers for three years, they can assess the outcomes of the producers who follow or ignore their recommendations and interventions. Students share recommendations and outcomes associated with their herd with the entire class. This allows students to learn how to establish best management practices through objective analysis of outcomes of recommended practices of all herds represented in the class. While a formal assessment of the course is needed, the students rate the program very high on evaluations.

The relationship of soil province to molecular characterization and phylogenetic analysis of Cryptosporidium parvum isolated from calves in Georgia.

Although Cryptosporidium spp. are found throughout the world and in multiple environmental conditions, few data are available that explore the possibility of an association between specific environmental parameters and the species or strain of Cryptosporidium. This study examines the potential association between a particular Cryptosporidium species/strain found in calves and soil provinces in Georgia, USA. Necropsy cases spanning the years 1996-2002 were tested. No significant differences (P=0.962, chi(2) test of homogeneity) between numbers of positive cases were noted among soil provinces. Phylogenetic analysis of the sequences for the PCR products revealed sequence similarity of the products with Cryptosporidium parvum strain C1. Although, clinical Cryptosporidiosis in calves was not found to be affected by soil province and may be caused by a single genotype, other genotypes may be responsible for subclinical infection and warrant further investigation.

The seroprevalence of Johne's disease in Georgia beef and dairy cull cattle.

Beef and dairy cattle serum samples, collected during 2000 at sale barns throughout Georgia, were obtained from the Georgia State Brucellosis Laboratory and were used to conduct a retrospective epidemiological study. Statistical samplings of 5,307 sera, from over 200,000 sera, were tested for antibodies to Mycobacterium avium ssp. paratuberculosis, (Johne's disease) using a commercial enzyme-linked immunosorbent assay test kit. An overall period seroprevalence in all classes of cattle tested was 4.73%. The period seroprevalence in dairy cattle was 9.58%, in beef cattle it was 3.95%, and in cattle of unknown breed it was 4.72%. It was concluded that the seroprevalence of Johne's disease in cull beef and dairy cattle in Georgia is economically significant.

Seroprevalence and comparison of isolates of endometriotropic bovine herpesvirus-4.

Sixty-eight cases of suppurative, ulcerative endometritis associated with Bovine Herpesvirus-4 (BHV-4) in postparturient dairy cows (62 Holsteins and 6 Jerseys, mean age 4.2 years) were confirmed by a combination of histopathology, fluorescent antibody assays, electron microscopic evaluation of uterus, and polymerase chain reaction (PCR). All cases occurred in the 3- to 28-day postpartum period, and histologic lesions among various cows were consistent when compared with postpartum interval. The endometrial lining epithelium was necrotic and ulcerated from 3 to 7 days postpartum, with only mild inflammation in the lamina propria and submucosa. From 1 to 4 weeks postpartum, the ulcers were confluent to diffuse. Epithelium was replaced by fibrinonecrotic, suppurative mats, resulting in severe bacterial pyometra by day 24. Seroprevalence to BHV-4 in one dairy with a history of 18 mortality cases was 36% (107 of 296). In a random sample of 8 cows from this herd, none had serologic titers in blood sampled 2 weeks prepartum, but 3 of 8 seroconverted with significant titers of 1:8 to 1:16 at 2 weeks postpartum. By 10 weeks postpartum, all 8 cows returned to negative serologic status. Two of 6 cats from the premises also had positive titers. Random serum samples taken from 480 dairy cattle at sale barns indicated 76 (16%) were positive by serum neutralization. Clinical signs, postparturient timing, and histologic lesions were very similar to those previously reported in Belgium with BHV-4. But sequence analysis of PCR products of the glycoprotein B region of 4 separate field isolates of endometriotropic BHV-4 suggests these field isolates were more closely related to the North American nonvirulent strain DN-599 than to the endometriotropic European strain V.

Congenital erythropoietic protoporphyria in a Limousin calf.

Bovine congenital erythropoietic protoporphyria is an uncommon genetic defect in Limousin and Blonde d'Aquitaine cattle that is characterized by severe photosensitization. Clinical signs include intense pruritus and exudative dermatitis involving the face, pinnae, and dorsal aspect of the thorax. Affected cattle have hematologic and serum biochemical values within reference ranges, and their teeth are normochromic. Definitive diagnosis of bovine congenital erythropoietic protoporphyria is accomplished by genetic testing. Affected cattle should be sent to a terminal market.