An enzyme-linked immunosorbent assay (elisa) for the detection of antibodies against babesia boris in cattle

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Skin test and tick immune status in susceptible and resistant cattle in Brazil

A cutaneous hypersensitivity test (CHT) was used to correlate host resistance to ticks and type of reaction elicited to unfed larval extract-ULE of the cattle tick Boophilus microplus in European and Indian cattle. Twenty calves were separated into four groups of five animals each: naïve or preinfested Indian or European cattle. CHT was induced by intradermal inoculation of 0.1 ml of ULE cattle tick B. microplus (50 μg protein) in the calf ear. Ear thickness was measured using calipers before and 10 min, 1, 2, 6, 18, 24, 48, 72, 96, and 144 h postinoculation (PI). Preinfested European calves showed only an immediate type reaction with maximum response (75% increase in ear thickness) at 10 min PI. On the other hand, preinfested Indian calves presented an immediate response with maximum reaction (70% increase in ear thickness) between 10 min and one hour PI, and a delayed type reaction at 72 h PI (60% increase in ear thickness). These results point out the crucial role of the cellular immune response of cattle in the expression of resistance to cattle tick B. microplus. Skin test might be useful in the ranking of cattle according to the susceptibility/resistance to ticks.

Araçatuba virus: A vaccinialike virus associated with infection in humans and cattle

We describe a vaccinialike virus, Araçatuba virus, associated with a cowpoxlike outbreak in a dairy herd and a related case of human infection. Diagnosis was based on virus growth characteristics, electron microscopy, and molecular biology techniques. Molecular characterization of the virus was done by using polymerase chain reaction amplification, cloning, and DNA sequencing of conserved orthopoxvirus genes such as the vaccinia growth factor (VGF), thymidine kinase (TK), and hemagglutinin. We used VGF-homologous and TK gene nucleotide sequences to construct a phylogenetic tree for comparison with other poxviruses. Gene sequences showed 99% homology with vaccinia virus genes and were clustered together with the isolated virus in the phylogenetic tree. Araçatuba virus is very similar to Cantagalo virus, showing the same signature deletion in the gene. Araçatuba virus could be a novel vaccinialike virus or could represent the spread of Cantagalo virus.

Foamy macrophages in the liver of cattle fed Brachiaria brizantha hay

Liver and lymph nodes injuries characterized by clusters of foamy macrophages, some of them containing birefringent crystals, were observed in cattle fed on Brachiaria brizantha hay. The cattle were from an experimental group poisoned with Senecio brasiliensis known to cause hepatic fibrosis and hepatocyte megalocytosis. One of the animals developed photosensitivity but the exact cause wasn't determined since both plants were fed. The foamy macrophages were present from the 30th d of feeding. Early appearance of these lesions may be particular to the animal specie used or due to the presence of both toxic plants.

Effects of vaccination against reproductive diseases on reproductive performance of beef cows submitted to fixed-timed AI in Brazilian cow-calf operations

The objectives were to assess incidence of pregnancy losses, associate this outcome with immunization programs against reproductive diseases, and evaluate the effects of vaccination against bovine herpesvirus-1 (BoHV-1), bovine viral diarrhea virus (BVDV), and Leptospira spp., on reproductive efficiency of Brazilian cow-calf operations. In experiment 1, 7614 lactating Nelore cows from 18 ranches were assigned to the same estrus synchronization and fixed-time AI protocol (ESFTAI; Days -11 to 0). Pregnancy status was determined with transrectal ultrasonography on Days 30 and 120 after AI. Pregnancy loss was deemed to have occurred when cows were pregnant on Day 30 but nonpregnant on Day 120. Incidence of pregnancy loss across all ranches was 4.1%; pregnancy losses were detected (P < 0.10) in 14 ranches but not detected (P > 0.11) in four ranches. Pregnancy loss was lower (P ≤ 0.02) in ranches that vaccinated against BoHV-1, BVDV, and Leptospira spp. compared with ranches that did not vaccinate, or only vaccinated against Leptospira spp. In experiments 2 and 3, lactating Nelore cows (N = 1950 and 2793, respectively) from ranches that did not have a history of vaccinating against reproductive diseases (experiment 2), or only vaccinated against Leptospira spp. (experiment 3), were assigned to the same ESFTAI used in experiment 1. Within each ranch, cows received (VAC) or not (CON) vaccination against BoHV-1, BVDV, and Leptospira spp. at the beginning of the ESFTAI (Day -11) and 30 days after (Day 41) AI. In experiment 2, VAC cows had greater (P ≤ 0.05) pregnancy rates compared with CON on Days 30 and 120. In experiments 2 and 3, pregnancy loss was reduced (P ≤ 0.03) in primiparous VAC cows compared with CON cohorts. In experiment 4, 367 primiparous, lactating Nelore cows previously vaccinated against Leptospira spp. were assigned to the same ESFTAI used in experiment 1. Cows received VAC, or the same vaccine 30 days before (Day -41) and at the beginning (Day -11) of the ESFTAI (PREVAC). Pregnancy rates on Days 30 and 120 were greater (P ≤ 0.05) in PREVAC cows compared with VAC cows. In conclusion, pregnancy losses affected reproductive and overall efficiency of Brazilian cow-calf operations, and might be directly associated with BoHV-1, BVDV, and Leptospira spp. infections. Hence, vaccinating cows against these pathogens, particularly when both doses are administered before fixed-time AI, improved reproductive performance in Brazilian cow-calf systems. © 2013 Elsevier Inc.

Babesia bovis infection in cattle in the southwestern Brazilian Amazon

The present study provides the first epidemiological data on infection with Babesia bovis in cattle raised in the southwestern Brazilian Amazon. Blood clot samples were filtered through nylon cloth before being submitted to DNA extraction. PCR and nested-PCR were applied to assess the frequency of infection with B. bovis in calves with ages from 4 to 12 months bred in 4 microregions each in the states of Rondônia and Acre. After the DNA was extracted from the samples, the infection in cattle was investigated by amplification of the rap1 gene from B. bovis. The DNA amplification results revealed a frequency of infection with B. bovis of 95.1% (272/286) in the samples from Rondônia and 96.1% (195/203) in those from Acre. The high frequency of B. bovis infection in the animals with ages from 4 to 12 months indicates a situation of enzootic stability in the regions studied. The infection rates are comparable to those detected by immunodiagnostic techniques in other endemic regions of Brazil. © 2012 Elsevier GmbH.

Persistent efficacy of 3.5% doramectin compared to 3.15% ivermectin against gastrointestinal nematodes in experimentally-infected cattle in Brazil

The present study aimed to evaluate the persistent efficacy of a 3.5% doramectin* (700μg/kg) formulation compared to 3.15% ivermectin** (630μg/kg) treatment, administered subcutaneously at a dose of 1mL/50kg body weight in cattle experimentally infected with gastrointestinal nematodes. Seventy-two male crossbred Holstein cattle that were negative for helminth infection were divided into nine groups. Treatments of 3.5% doramectin (Groups 2, 4, 6 and 8) and 3.15% ivermectin (Groups 3, 5, 7 and 9) were administered on days 49, 42, 35 and 28 prior to challenge with infectious nematode larvae (L3). Animals in the control group (Group 1) received saline solution on day 49 before challenge. Beginning on day zero, each animal received 50mL orally of a mixed culture containing approximately 3,000 third stage larvae (L3) of Haemonchus (60%), Oesophagostomum (20%), Cooperia (15%) and Trichostrongylus (5%) for seven consecutive days, resulting in a total challenge of 21,000 larvae/animal. Due to the large number of cattle, autopsies were performed between days 28 and 35 after the last day of inoculation. The formulation containing doramectin (700 mcg/kg) achieved persistent efficacy against H. placei and C. punctata for 49 and 35days, respectively. The persistent efficacy of ivermectin (630 mcg/kg) against H. placei lasted for 49days, but this treatment was ineffective against C. punctata. Both formulations demonstrated persistent efficacy against T. axei for 49days. The persistent efficacy of doramectin (700 mcg/kg) and ivermectin (630 mcg/kg) lasted for 49 and 42days against O. radiatum, respectively. © 2012 Elsevier Ltd.

Experimental infection of bovines with oocysts of Toxoplasma gondii

Calves aged 3 mth were readily infected with oocysts and cysts of Toxoplasma gondii administered by the oral route. Fever, respiratory distress, nasal discharge, and hyperemia of the conjunctivas were the most significant clinical signs noted in the infected animals. Parasitemia was demonstrated in all infected calves. It occurred on different days and up to 62 days after the infection. Toxoplasma was demonstrated in tissues of all infected calves, and the organ most frequently parasitized was the lymph node. Parasitism of the retina was demonstrated in 2 calves. All infected animals had antibody against T. gondii in their serum. The Sabin-Feldman dye test and the indirect immunofluorescent test were both useful in detecting antitoxoplasma antibody.

A comparative study of the efficiency of a pro-biotic and the anti-K99 and anti-A14 vaccines in the control of diarrhea in calves in Brazil.

A total of 99 pregnant cows were divided into eight groups submitted to the following treatments: group I (n = 29) consisted of unvaccinated cows whose calves did not receive a probiotic and was used as control. Group II (n = 10) consisted of vaccinated cows whose calves did not receive a probiotic. Groups III, IV and V (n = 10 neach) consisted of vaccinated cows whose calves received a probiotic for 5, 15 and 30 days, respectively. Groups VI, VII and VIII (n = 10 each) consisted of unvaccinated cows whose calves received a probiotic for 5, 15 and 30 days, respectively. Each animal in the vaccinated groups received two 5.0 ml vaccine doses containing pili K99 and A14 of Escherichia coli by the subcutaneous route. The probiotic containing Lactobacillus acidophilus at the dose of 2.0 x 10(8) live cells in 250 ml milk, was administered orally. All animals were observed clinically and bacteriologically and anti-K99 and anti-A14 antibody titers were determined in serum and colostrum. Mean calf weight was measured at birth and at 30 days of age. The results showed that a combination of the vaccine with the probiotic administered for 15 and 30 days was the most efficient treatment for the control of diarrhea.

Use of lytic bacteriophage for control of experimental Escherichia coli septicemia and meningitis in chickens and calves

A lytic bacteriophage, which was previously isolated from sewage and which attaches to the K1 capsular antigen, has been used to prevent septicemia and a meningitis-like infection in chickens caused by a K1+ bacteremic strain of Escherichia coli. Protection was obtained even when administration of the phage was delayed until signs of disease appeared. The phage was able to multiply in the blood. In newly borne colostrum-deprived calves given the E. coli orally, intramuscular inoculation of phage delayed appearance of the bacterium in the blood and lengthened life span. With some provisos there is considerable potential for this approach to bacterial-disease therapy.

Detection of leptospires in bovine semen by polymerase chain reaction

Objective: In view of the considerable importance of venereal transmission of bovine leptospirosis, the objective of the present study was to compare the polymerase chain reaction (PCR), culture/isolation and serology to detect leptospire infection in bovine semen. Design: Blood for serologic examination and semen for bacterial culture and PCR were collected from 20 bulls at artificial insemination centres in Brazil. Each animal was sampled twice for serology. Result: Forty-five percent (9/20) of the serum samples collected showed agglutinin titers to serovar hardjo in the first sample and 25% (5/20) had agglutinin titers to serovar hardjo in the second sample. Eighty percent (16/20) of semen samples were positive by PCR. Leptospires could not be isolated from any of the semen samples examined. Conclusion: Polymerase chain reaction can be a method of great potential for the detection of leptospires at artificial insemination centres.