Aspectos clínicos e epidemiológicos das infecções por Babesia bovis, Babesia bigemina e Anaplasma marginale em bezerros da raça Nelore no Estado de São Paulo

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Inherited hemoglobin disorders in an Afro-Amazonian community: Saracura

ABSTRACT: The most common hemoglobinopathies, viz, hemoglobins S and C, and α-and β-thalassemias, were investigated through the molecular screening of 116 subjects from the community of Saracura, comprising fugitive African slaves from farms of the municipality of Santarém, in the west of Pará State, Brazilian Amazon. The observed frequency of the HBB*S gene (0.9%) was significantly lower than that encountered in other Afro-derived communities in the region. Concomitantly, the absence of the HBB*C allele has been reported for most of the Afro-Amazonian communities thus far studied. As remnant populations of quilombos are generally small, the heterogeneous distribution of HBB*S and HBB*C alleles among them is probably due to genetic drift and/or founder effect. The observed frequency of 3.7 kb deletion in Saracura (8.5%) was consistent with the African origin of the population, with a certain degree of local differentiation and admixture with individuals of Caucasian ancestry, placed in evidence by the occurrence of - -(MED) deletion (1.2%), a common mutation in Mediterranean regions. As regards f-thalassemia, among the seven different mutations found in Saracura, three βºand two β+ mutations were of Mediterranean origin, and two β+ of African. Thus, only 28% of the local β-thalassemia mutations found in Saracura were of African origin.

Origin of the hemoglobin S gene in a northern Brazilian population: the combined effects of slave trade and internal migrations

Com o objetivo de investigar a origem da mutação bS na população da região norte do Brasil, foram analisados polimorfismos de DNA no complexo de genes b da hemoglobina em 30 pacientes com anemia falciforme na população de Belém, a capital do Estado do Pará. Sessenta e sete por cento dos cromossomos bS analisados apresentaram o haplótipo Bantu, 30% o haplótipo Benin e 3% o haplótipo Senegal. A origem da mutação bS na população de Belém, estimada de acordo com a distribuição de haplótipos, não está de acordo com a esperada com base em dados históricos sobre o tráfico de escravos para a região norte, os quais indicam uma reduzida contribuição de escravos da região do Benin. Essas diferenças podem ser atribuídas ao tráfico interno de escravos, bem como ao posterior fluxo de populações imigrantes, particularmente de nordestinos. A distribuição de haplótipos em Belém não difere significativamente da observada em outras regiões brasileiras, muito embora os dados históricos sugiram que a maioria dos escravos procedentes da região do Atlântico-Oeste africano, onde predomina o haplótipo Senegal, foi trazida para o norte do Brasil, enquanto que o nordeste (Bahia, Pernambuco e Maranhão) recebeu o maior contingente de escravos oriundos da região centro-oeste africana, onde o haplótipo Benin é o mais comum. Nós sugerimos que as diferenças regionais quanto à procedência dos escravos africanos também foram modificadas pelo tráfico de escravos estabelecido entre as diferentes regiões brasileiras e posteriormente pelos movimentos migratórios.

Direct detection of Mycobacterium tuberculosis complex in bovine and bubaline tissues through nested-PCR

Post-mortem bacterial culture and specific biochemical tests are currently performed to characterize the etiologic agent of bovine tuberculosis. Cultures take up to 90 days to develop. A diagnosis by molecular tests such as PCR can provide fast and reliable results while significantly decreasing the time of confirmation. In the present study, a nested-PCR system, targeting rv2807, with conventional PCR followed by real-time PCR, was developed to detect Mycobacterium tuberculosis complex (MTC) organisms directly from bovine and bubaline tissue homogenates. The sensitivity and specificity of the reactions were assessed with DNA samples extracted from tuberculous and non-tuberculous mycobacteria, as well as other Actinomycetales species and DNA samples extracted directly from bovine and bubaline tissue homogenates. Regarding the analytical sensitivity, DNA of the M. bovis AN5 strain was detected up to 1.5 pg by nested-PCR, whereas DNA of M. tuberculosis H37Rv strain was detected up to 6.1 pg. The nested-PCR system showed 100% analytical specificity for MTC when tested with DNA of reference strains of non-tuberculous mycobacteria and closely-related Actinomycetales. A clinical sensitivity level of 76.7% was detected with tissues samples positive for MTC by means of the culture and conventional PCR. A clinical specificity of 100% was detected with DNA from tissue samples of cattle with negative results in the comparative intradermal tuberculin test. These cattle exhibited no visible lesions and were negative in the culture for MTC. The use of the nested-PCR assay to detect M. tuberculosis complex in tissue homogenates provided a rapid diagnosis of bovine and bubaline tuberculosis.

Inefficacy of albendazole sulphoxide and ivermectin for the treatment of bovine parasitic otitis caused by rhabditiform nematodes

O objetivo deste estudo foi avaliar a eficácia do sulfóxido de albendazol administrado oralmente e da ivermectina "pour-on" no tratamento da otite parasitária bovina causada por nematóides rhabditiformes. Dezoito vacas Gir apresentando otite clínica foram divididas em três grupos de seis animais cada. O primeiro não recebeu tratamento (grupo controle). O segundo foi tratado com ivermectina "pour-on" a 0,05% na dose de 500µg/kg de peso vivo. O terceiro grupo foi tratado com sulfóxido de albendazol oral a 6% na dose 6,0mg/kg. Os condutos auditivos de todos os animais foram reexaminados nos dias 7 e 21 pós-tratamento. Os animais do grupo controle permaneceram infectados nos dias de observação. O tratamento com ivermectina não demonstrou eficácia alguma para os dias 7 e 21 pós-tratamento. O tratamento com sulfóxido de albendazol obteve 16,7 e 25% de eficácia nos dias 7 e 21, respectivamente. Mais estudos são necessários para determinação de tratamentos eficazes para tal doença parasitária, especialmente através de vias alternativas de administração, por causa de seu significante impacto na criação de Bos taurus indicus nas Regiões Tropical e Subtropical.

Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Molecular Evaluation of Developmental Competence of Oocytes Collected In Vivo from Buffalo and Bovine Heifers during Winter and Summer

Buffaloes and bovines are polyestrous and seasonal or annual livestock, respectively, that show reduced fertility during heat stress. To investigate whether reduced fertility is related to oocyte competence in both species, immature oocytes from buffalo and bovine heifers were collected during winter and summer and subjected to molecular analyses. In each season, heifers of both species had their follicular wave emergence synchronized with a standard protocol (Ferreira et al., 2011). Before being subjected to ovum pick up (OPU), cutaneous (CT; degrees C) and rectal (RT; degrees C) temperatures and respiratory rate (RR; breaths/min) were measured. Oocytes' RNA was extracted to evaluate the expression of target genes related to mtDNA replication/transcription (PPARGC1A, TFAM and MT-CO1), apoptosis (BAX and BCL2) and HS (HSP90AA1 and HSPA1AB). ACTB, HIST1H2AG and GAPDH were initially chosen as housekeeping genes. In buffaloes, CT (35.0 +/- 0.4 vs 23.8 +/- 0.5), RT (38.7 +/- 0.1 vs 38.0 +/- 0) and RR (21.3 +/- 1.2 vs 15.4 +/- 1.1) were higher during summer than winter. However, in bovine heifers, RT (38.7 +/- 0.1 vs 38.6 +/- 0.1) and RR (44.8 +/- 1.5 vs 40.6 +/- 1.5) were similar in both seasons, while CT (31.6 +/- 0.3 vs 30.2 +/- 0.3) was increased during summer. Reduced expression of ACTB, HIST1H2AG and GAPDH was evidenced during summer, disqualifying them as housekeeping genes. Similarly, the expression of all target genes was reduced during summer in oocytes of both species. In summary, physiological responses to heat stress seem to be more intense in buffalo than bovine heifers. However, in both species, negative effects of heat stress upon oocyte quality occur at the molecular level and affects genes related to several biological functions.

Cryopreservation of bovine spermatozoa from the epididymal tail using conventional and automated methods

Cryopreservation of sperm is important to preserve the gerrnplasm from animals of genetic value, which can die unexpectedly. This study compares conventional and automated methods of cryopreservation of spermatozoa obtained from the epididymis of bulls post-mortem. Twenty-two epididymides were obtained from a commercial slaughterhouse. Spermatozoa were collected from the tail of the epididymis using the retrograde flow technique. Thus, the samples, which were diluted in 10 ml of extender without glycerol (Botubov (R) I, Botupharma, Botucatu, SP, Brazil), were evaluated on motility, sperm vigor, structural and functional (swelling hypoosmotic test) membrane integrity, mitochondrial activity, sperm viability and ADN fragmentation. The samples were divided into two aliquots and diluted in extender with glycerol (Botubov (R) II, Botupharma, Botucatu, SP, Brazil) at a concentration of 50x10(6) motile sperm/0.5 French straws. One sample was frozen by the conventional method (4 hours at 5 degrees C, in a refrigerator and 20 min in nitrogen vapor) and the other by the automated method (Cryogen (R) Dualflex, Neovet, Uberaba, MG, Brazil). The parameters were higher in all the tests of fresh sperm samples, with the exception of the swelling hypoosmotic test, which showed no significant difference when the results were compared with sperm frozen by the conventional method. The average motility of fresh spermatozoa was 74%, and conventional and automated averages were 29 and 25%, respectively. Therefore, although cryopreservation techniques reduce sperm quality parameters, the viability of the sperm is maintained, and these methods can be used to preserve sperm.

Isolate((R)) and Optiprep((R)) minigradients as alternatives for sperm selection in bovine in vitro embryo production

The objective of this study was to evaluate alternatives in small volumes to conventional gradient of Percoll((R)) on semen quality, in vitro embryo production, sex ratio and embryo survival after vitrification. Thawed semen was randomly allocated to one of four density gradient selection methods: (1) conventional Percoll((R)) (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), and (4) MiniOptiprep (MO). Sperm kinetics and quality were evaluated. Use of P, MP and MI gradients did not affect sperm motility (P > 0.05). However, there was a decrease in total and progressive sperm motility in MO (70.8 and 51.3% vs. 87.3 and 69.5% for P; 87.3 and 73% for MP; 92.3 and 78.8% for MI; P < 0.05). The MO had lower membrane integrity compared with P, MP and MI (39.7 vs. 70.5, 72.3, 63.8%, respectively, P < 0.05). The percentage of blastocysts produced was higher in MI than in MP and MO (21.1 vs. 16.1 and 16.9%, P < 0.05) and similar to P (18.4%; P > 0.05). Sex ratio and embryo survival after vitrification were similar among groups (P > 0.05). Semen selected by Isolate and Optiprep gradient, at the concentrations and small volumes used, demonstrated similar characteristics and in vitro embryo production to conventional Percoll((R)) gradient.

Live High-Train High increases hemoglobin mass in Olympic swimmers

This study tested whether 3-4 weeks of classical Live High-Train High (LHTH) altitude training increases swim-specific VO2max through increased hemoglobin mass (Hb(mass)).Ten swimmers lived and trained for more than 3 weeks between 2,130 and 3,094 m of altitude, and a control group of ten swimmers followed the same training at sea-level (SL). Body composition was examined using dual X-ray absorptiometry. Hb(mass) was determined by carbon monoxide rebreathing. Swimming VO2peak was determined and swimming trials of 4 x 50, 200 and 3,000 m were performed before and after the intervention.Hb(mass) (n = 10) was increased (P < 0.05)after altitude training by 6.2 +/- A 3.9 % in the LHTH group, whereas no changes were apparent in the SL group (n = 10). Swimming VO2peak was similar before and after training camps in both groups (LHTH: n = 7, SL: n = 6). Performance of 4 x 50 m at race pace was improved to a similar degree in both groups (LHTH: n = 10, SL: n = 10). Maximal speed reached in an incremental swimming step test (P = 0.051), and time to complete 3,000 m tended (P = 0.09) to be more improved after LHTH (n = 10) than SL training (n = 10).In conclusion, 3-4 weeks of classical LHTH is sufficient to increase Hb(mass) but exerts no effect on swimming-specific VO2peak. LHTH may improve performance more than SL training.

Programmed cell death-associated gene transcripts in bovine embryos exposed to bovine Herpesvirus type 5

In vitro-produced bovine embryos become infected after exposure to bovine Herpesvirus type 5 (BoHV-5), yet no changes in developmental rates, mitochondrial activity and inhibition of apoptosis are detected in comparison to unexposed embryos. Thus, the aim of the present study was to assess the transcription of mitochondria-mediated apoptosis genes using TaqMan real-time polymerase chain reaction. Transcripts of mcl-1, caspase-2, -3, Apaf-1 and Bax genes were measured after exposure to BoHV-5 in vitro. Mitochondrial dehydrogenase activity was evaluated by MIT test and compared between groups of exposed and unexposed embryos, at day 7 of development. The rate of oocyte maturation was assessed by the extrusion of the first polar body. In summary, BoHV-5 exposed embryos retained their viability, mitochondrial dehydrogenase activity and displayed up-regulation of transcription of survival mcl-1 gene and down-regulation of Bax transcription in relation to mitochondria-mediated pathway which might improve embryo viability. These findings demonstrate that BoHV-5 exposed embryos maintain their viability and mitochondrial dehydrogenase activity with no compromise of embryos produced in vitro. (c) 2013 Elsevier Ltd. All rights reserved.