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

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Associated and independent comparisons between the two largest follicles preceding follicle deviation in cattle

Follicle diameters and concentrations of follicular fluid factors were studied in the two largest follicles (F1 and F2) using F1 diameters in increments of 0.2 mm (equivalent to 4 h intervals) and extending from 7.4 to 8.4 mm (12 heifers in each of 6 groups). Changes were compared between follicles using the F2 associated with each F1-diameter group. Diameter deviation began in the 8.2-mm group as indicated by a greater (P < 0.05) diameter difference between F1 and F2 in the 8.4-mm group than in the 8.2-mm group. In the 8.0-mm group, estradiol concentrations began to increase (P < 0.05) differentially in F1 versus F2, and free insulin-like growth factor-1 (IGF-1) began to decrease differentially in F2 (P < 0.06). Combined for F1 and the associated F2, activin-A concentrations increased (P < 0.05) between the 7.6- and 8.2-mm groups and then decreased (P < 0.05). Results supported the hypothesis that estradiol and free IGF-1 concentrations simultaneously become higher in F1 than in the associated F2 by the beginning of diameter deviation. Results did not support the hypothesis that a transient elevation in activin-A is present in F1 but not in the associated F2 at the beginning of the estradiol and IGF-1 changes; instead, a mean transient elevation in activin-A occurred at this time only when data for the two follicles were combined. Comparisons between F1 and F2 also were made by independently grouping F2 and using diameter groups at 0.2-mm increments for F2 as well as for F1. In the diameter groups common to F1 and F2 (7.4, 7.6, 7.8, and 8.0 mm) there was a group effect (P < 0.003) for estradiol involving an increase (P < 0.05) beginning at the 7.6-mm group averaged over F1 and F2. For free IGF-1 concentrations, a fluctuation (a significant increase followed by a significant decrease) occurred independently in F1 between the 7.4-to 7.8-mm groups and independently in F2 between the 7.0- to 7.4-mm groups.

Interval between first dose and booster affected antibody production in cattle vaccinated against rabies

In this study, we compared the levels of neutralizing antibodies induced by inactivated rabies vaccine in cattle by using three alternative immunization procedures. Forty-five bovines (breed nelore) were then organized in three groups (A, B and C, with 15 animals/group). Group A received only one vaccine dose at day zero and Group B received the first dose at day zero and then another dose at day 30 (early booster). Group C was also immunized with two doses; however, the booster was postponed until day 180 after the first dose (delayed booster). Blood samples were withdrawn at days zero (before the first dose) and 30, 210, 390, and 540 after the beginning of immunization and the antibody titers were evaluated by mouse neutralization test. The protocol used to immunize Group C (booster at day 180) was clearly more efficient. In this group, antibody levels were higher and also remained higher for longer periods in comparison with the other two groups. These results show that booster timing significantly affected antibody levels. Therefore, programs addressed to control this disease in cattle should consider not only the use of a booster but also its administration time.

Selenium supplementation enhances weight gain in cattle

To evaluate the effect of selenium on cattle growth in a continuous grazed pasture system. Selenium was added to protein-mineral salt and given to male, around-12-month old, non-castrated Nellore calves. Animals were randomly and equally distributed into 4 groups (15 calves/group) which received supplementation containing 0, 3.6, 5.4 or 6.4 mg selenium/animal/day (groups Gc, G 3.6, G 5.4 and G 6.4, respectively). The animals were weighed on day zero and 120, and the pasture forages were collected at day 0 for chemical analyses. Weight gain was higher in cattle from G 5,4 (45.58%) than in the other groups, and higher in G 3,6 (24.97%) and G 6,4 (22.67%) than in Gc. The supplementation with 5.4 mg selenium/animal/day enhanced weight gain in cattle fed on selenium poor diet (0.04 mg Se/kg dry matter).

Effect of superstimulatory treatments on the expression of genes related to ovulatory capacity, oocyte competence and embryo development in cattle.

Multiple ovulation (superovulation) and embryo transfer has been used extensively in cattle. In the past decade, superstimulatory treatment protocols that synchronise follicle growth and ovulation, allowing for improved donor management and fixed-time AI (FTAI), have been developed for zebu (Bos indicus) and European (Bos taurus) breeds of cattle. There is evidence that additional stimulus with LH (through the administration of exogenous LH or equine chorionic gonadotrophin (eCG)) on the last day of the superstimulatory treatment protocol, called the 'P-36 protocol' for FTAI, can increase embryo yield compared with conventional protocols that are based on the detection of oestrus. However, inconsistent results with the use of hormones that stimulate LH receptors (LHR) have prompted further studies on the roles of LH and its receptors in ovulatory capacity (acquisition of LHR in granulosa cells), oocyte competence and embryo quality in superstimulated cattle. Recent experiments have shown that superstimulation with FSH increases mRNA expression of LHR and angiotensin AT(2) receptors in granulosa cells of follicles >8 mm in diameter. In addition, FSH decreases mRNA expression of growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) in oocytes, but increases the expression of both in cumulus cells, without diminishing the capacity of cumulus-oocyte complexes to generate blastocysts. Although these results indicate that superstimulation with FSH is not detrimental to oocyte competence, supplementary studies are warranted to investigate the effects of superstimulation on embryo quality and viability. In addition, experiments comparing the cellular and/or molecular effects of adding eCG to the P-36 treatment protocol are being conducted to elucidate the effects of superstimulatory protocols on the yield of viable embryos.

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.

Serological study of vaccinia virus reservoirs in areas with and without official reports of outbreaks in cattle and humans in São Paulo, Brazil

Vaccinia virus (VACV), the etiological agent of an exanthematic disease, has been associated with several bovine outbreaks in Brazil since the end of the global vaccination campaign against smallpox. It was previously believed that the vaccine virus used for the WHO global campaign had adapted to an unknown wild reservoir and was sporadically re-emerging in outbreaks in cattle and milkers. At present, it is known that Brazilian VACV is phylogenetically different from the vaccinia virus vaccinal strain, but its origin remains unknown. This study assessed the seroprevalence of orthopoxviruses in domestic and wild animals and farmers from 47 farms in three cities in the southwest region of the state of São Paulo with or without official reports of outbreaks in cattle or humans. Our data indicate a low seroprevalence of antibodies in wild animals and raise interesting questions about the real potential of wild rodents and marsupials as VACV reservoirs, suggesting other routes through which VACV can be spread. © 2013 The Author(s).

Effects of FSH on the expression of receptors for oocyte-secreted factors and members of the EGF-like family during in vitro maturation in cattle

FSH induces expansion of bovine cumulus-oocyte complexes (COCs) in cattle, which can be enhanced by oocyte-secreted factors (OSFs). In this study it was hypothesised that FSH stimulates COC expansion in part from direct stimulation of the epidermal growth factor (EGF)-like ligands amphiregulin (AREG), epiregulin (EREG) and betacellulin (BTC), but also in part through regulation of OSFs or their receptors in cumulus cells. Bovine COCs were cultured in defined medium with graded doses of FSH. In the absence of FSH, COCs did not expand. FSH caused cumulus expansion, and increased the abundance of AREG and EREG mRNA in a time- and dose-dependent manner, but decreased BTC mRNA levels. FSH had modest stimulatory effects on the levels of mRNA encoding the bone morphogenetic protein 15 (BMP15) receptor, BMPR1B, in cumulus cells, but did not alter mRNA expression of the growth and differentiation factor 9 (GDF9) receptor, TGFBR1. More interestingly, FSH dramatically stimulated levels of mRNA encoding two receptors for fibroblast growth factors (FGF), FGFR2C and FGFR3C, in cumulus cells. FSH also stimulated mRNA expression of FGFR1B, but not of FGFR2B in cumulus cells. Based on dose-response studies, FGFR3C was the receptor most sensitive to the influence of FSH. This study demonstrates that FSH stimulates the expression of EGF-like factors in bovine cumulus cells, and provides evidence that FSH differently regulates the expression of distinct receptors for OSFs in cumulus cells. © CSIRO 2013.

Frequency of antibodies to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burdgorferi in cattle from the northeastern region of the state of Pará, Brazil

A babesiose, a anaplasmose e a tripanossomose são enfermidades relevantes, potencialmente causadoras de morbidade em bovinos, levando a perdas econômicas. A borreliose assume importância como zoonose potencial. O objetivo desse estudo foi determinar, por meio do ensaio de imunoadsorção enzimática (ELISA), a freqüência de anticorpos para Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax e Borrelia burgdorferi em bovinos da região nordeste do Estado do Pará, Brasil. Amostras de soro de 246 vacas dos municípios de Castanhal e São Miguel do Guamá foram usadas. ELISAs com antígeno bruto foram utilizados para detector anticorpos contra todos os agentes, exceto para A. marginale, para o qual um ELISA indireto com proteína principal de superfície 1a (MSP1a) foi usado. As freqüências de bovinos soropositivos foram: B. bigemina - 99,2%; B. bovis - 98,8%; A. marginale - 68,3%; T. vivax - 93,1% and B. burgdorferi -54,9% As freqüências de bovinos soropositivos para B. bovis e B. bigemina sugerem uma alta taxa de transmissão desses organismos por carrapatos, na região estudada, a qual pode ser classificada com sendo de estabilidade enzoótica para os hemoparasitos. A baixa freqüência de bovinos soropositivos para A. marginale pode ser atribuída a uma menor sensibilidade do ELISA com antígeno recombinante, ou uma menor taxa de inoculação da riquétsia pelos carrapatos, quando comparada àquelas observadas para Babesia sp. A alta freqüência de bovinos soropositivos para T. vivax indica que esse hemoprotozoário é prevalente em rebanhos do nordeste do Estado do Pará. O percentual de animais com anticorpos homólogos para B. burgdorferi indica a presenças deste espiroquetídeo transmitido por carrapatos na população de bovinos da região estudada.

Development and assessment of a latex agglutination test based on recombinant MSP5 to detect antibodies against Anaplasma marginale in cattle

The recombinant protein MSP5 has been established as an important antigen for serological diagnosis of Anaplasma marginale by enzyme-linked immunosorbent assay (ELISA). However, due to the high cost of specialized equipment, this technique is not accessible to all laboratories, especially in developing countries in areas where the disease is endemic. The present study describes the standardization of a latex agglutination test (LAT) to detect antibodies against A. marginale based on recombinant MSP5. Compared with indirect enzyme-linked immunosorbent assay (iELISA), the relative sensitivity and specificity of the LAT were 95.21% and 91.86% respectively, with an almost perfect agreement between tests (kappa index = 0.863). These results can be considered important for the serological diagnosis of A. marginale, as they indicate that the test represents a rapid and low cost alternative to ELISA.

Methods of inducing conditioned food aversion to Baccharis coridifolia (mio-mio) in cattle

Foram realizados três experimentos para determinar a eficácia de vários métodos de aversão a Baccharis coridifolia em bovinos: administração oral forçada de 0,5g kg^-1 de peso vivo de B. coridifolia fresca; inalação forçada da fumaça proveniente da queima de B. coridifolia e esfregação da planta no focinho e na boca dos animais; e introdução dos animais em pastagens com baixa infestação por B. coridifolia. Os resultados demonstraram que os animais forçados a ingerir pequenas doses tornaram-se fortemente avertidos, quando introduzidos nos piquetes entre 23 a 26 horas após a aversão. Entretanto, bovinos introduzidos nos piquetes após 1 a 10 horas não foram totalmente avertidos. Inalação da fumaça de B. coridifolia e esfregação da planta no focinho e boca dos animais não foram eficientes para produzir aversão. A introdução de bovinos em piquetes com aproximadamente 1% de B. coridifolia foi eficiente quando os animais permaneceram cinco meses na área, mas não quando ficaram apenas 60 horas, pois os bovinos precisam de tempo para aprender a evitar a planta.

Lead poisoning in cattle and chickens in the state of Pará, Brazil

O presente estudo descreve a ocorrência de intoxicação por chumbo em bovinos e galinhas no Pará, Brasil. Em um lote composto de 80 bezerros de um rebanho leiteiro, 10 animais ficaram doentes e nove morreram, e um animal se recuperou após ser removido do piquete. Após a inspeção deste piquete, foi observada a presença de baterias de caminhões usados para armazenar a energia captada por painéis solares. Os sinais clínicos observados nos bezerros incluíam dificuldade respiratória, corrimento nasal, salivação excessiva, opacidade da córnea, pressão da cabeça contra objetos e decúbito. As galinhas tinham diminuída oviposição e os ovos produzidos eram com cascas malformadas ou tinham a casca mais fina. Os achados de necropsia e as alterações histopatológicas observadas nos bovinos eram de pouco significado, com exceção de um animal que mostrou leve astrocitose no córtex cerebral. Em uma das galinhas, na histopatologia renal observou-se leve necrose tubular aguda multifocal. As concentrações de chumbo médios nos fígados e rins dos bovinos eram 93,91mg/kg e 209,76mg/kg, respectivamente, e a concentração média no fígado de galinhas foi 105,02mg/ kg. Concluiu-se que a fonte de contaminação por chumbo nesses bezerros e galinhas eram placas de bateria de caminhão, aos quais os animais tiveram acesso na pastagem.

Degenerative joint disease in cattle and buffaloes in the Amazon region: a retrospective study

Foi realizado um estudo retrospectivo sobre os aspectos epidemiológicos e clínico-patológicos em bovinos e búfalos com doença articular degenerativa (DAD) no estado do Pará, Brasil. Durante os anos de 1999 a 2014 foram avaliados 11 bovinos e 24 bubalinos. Todos os animais atendidos com suspeita clínica de DAD foram submetidos a exame clínico do sistema locomotor. Foram necropsiados sete bovinos e oito bubalinos com sinais clínicos da enfermidade. Os sinais clínicos comuns observados em ambas as espécies foram claudicação crônica, andar rígido, alterações posturais, crepitações audíveis no membro acometido, decúbito prolongado, dificuldade para levantar, e emagrecimento progressivo. As lesões articulares evidenciadas na necropsia consistiram em irregularidade da superfície articular, presença de erosão na cartilagem articular e no tecido ósseo subjacente, proliferação de tecido ósseo periarticular com formação de osteófitos. Tanto nos bovinos como nos bubalinos as articulações mais acometidas foram as dos membros posteriores. Nos bubalinos, possivelmente o principal fator predisponente ao surgimento de DAD foi à deficiência de fósforo, ao contrário dos bovinos, nos quais os defeitos de conformação anatômica dos membros posteriores, traumas crônicos em virtude da atividade exercida, como a coleta de sêmen e a idade avançada, foram o que, possivelmente, contribuíram para surgimento da enfermidade.

Expression of receptors for BMP15 is differentially regulated in dominant and subordinate follicles during follicle deviation in cattle

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Assessing resistance of ivermectin and moxidectin against nematodes in cattle naturally infected using three different methodologies

The objective of this study was to evaluate the accuracy of the faecal egg count reduction test (FECRT) and the faecal egg count efficacy test (FECET) to assess the resistance status of ivermectin (630 mu g/g) and moxidectin (200 mu g/kg), using the controlled efficacy test as a reference, and whether the results of the EPG are equivalent to the efficacy results from the parasitological necropsies. Two experiments were conducted. The results demonstrate that it was not possible to demonstrate that the EPG values were equivalent with the ivermectin and moxidectin efficacy obtained by parasitological necropsies, mainly if the phenomenon of parasites resistance is not advanced in a determined field population. Maybe the FECET technique would be possibly better than the FECRT. The high anthelmintic efficacy of 200 mu g/kg moxidectin, in naturally infected cattle, against field population of nematodes that are resistant to 630 mu g/kg ivermectin, was observed in this study. (C) 2013 Elsevier Ltd. All rights reserved.