Genome association analysis for pregnancy status following parturition in crossbred beef cattle / Estudo de associação genômica de prenhez pós-parto em vacas de corte

The aim of this study was to evaluate genetic diversity of nine molecular markers, six short tandem repeats - STRs (BM4325, BMS3004, ILSTS002, IDVGA51, HEL5, AFZ1) and three single nucleotide polymorphisms (SNPs; LepSau3A1 A-B, LepSau3A1 1-2, and FSHRAlu1), linked to genes involved in reproductive function and their possible effect on reproductive performance. For this purpose, 81 crossbred beef cows were used in this study. The animals were classified into two groups (fertile and sub-fertile cows) based on their pregnancy status after two breeding seasons. High genetic diversity level was observed highlighted by the polymorphic content information ranging 0.23 to 0.87 and expected heterozygosity from 27 to 89%, with an average of 62%. Alleles BM4325 103, BMS3004 129, ILSTS002 137, IDVGA51 177, LEPSau3A1 A, LEPSau3A1 1, HEL5 149, AFZ1 119 and FSHRAlu1 G presented high frequencies. Two STRs (IDVGA51 and ILSTS002), linked to Leptin and LH genes, respectively, were associated to reproductive performance. These data support previous findings suggesting the potential use of IDVGA51 and ILSTS002 STRs for reproductive performance selection.

Foi avaliada a diversidade genética de nove marcadores moleculares, dos quais seis do tipo short tandem repeats - STR (BM4325, BMS3004, ILSTS002, IDVGA51, HEL5, AFZ1) e três do tipo single nucleotide polymorphisms - SNPs (LepSau3A1 A-B, LepSau3A1 1-2 e FSHRAlu1), ligados a genes envolvidos na reprodução e seus efeitos na performance reprodutiva. Foram examinadas amostras de sangue de 81 vacas sem raça definida, os animais foram classificados em dois grupos (vacas férteis e subférteis) baseado nas taxas de prenhez de duas estações reprodutivas. Alto nível de diversidade genética foi observado, revelando alto conteúdo de informação polimórfica, variando de 0,23 a 0,87 e heterozigosidade esperada de 27 a 89% com 62% em média. Os alelos mais frequentes foram BM4325 103*, BMS3004 129*, ILSTS002 137*, IDVGA51 177*, LEPSau3A1 A, LEPSau3A1 1, HEL5 149*, AFZ1 119* e FSHRAlu1 G. Os marcadores IDVGA51 e ILSTS002, ligados aos genes da leptina e LH, respectivamente, foram associados a performance reprodutiva. Esses dados suportam achados prévios que sugerem o potencial uso desses marcadores na seleção de animais com maior performance reprodutiva.

Isolamento de Salmonella Braenderup em arara-azul (Anodorhynchus hyacinthinus) / Salmonella Braenderup isolation in Hyacinth macaw (Anodorhynchus hyacinthinus)

O monitoramento sanitário de populações de arara-azul (Anodorhynchus hyacinthinus) de vida livre visa a permitir ajustes de manejo em ambiente natural alterado. Para avaliar a ocorrência de Salmonella spp. em filhotes dessa espécie, foram coletados swabs de cloaca no Pantanal de Miranda, Mato Grosso do Sul (MS). Uma colônia morfológica e bioquimicamente compatível com Salmonella spp. foi isolada e sorotipada como Salmonella Braenderup. Devido ao alto potencial zoonótico desse microrganismo, é importante o controle sanitário de psitacídeos em vida livre. Na literatura pesquisada não foi encontrado relato sobre o isolamento dessa bactéria em arara-azul, tanto em vida livre, como em cativeiro.

The sanitary monitoring of free-living Hyacinth macaw (Anodorhynchus hyacinthinus) allows adjusting the management in altered natural habitat. To evaluate the occurrence of Salmonella spp. it was collected cloacal swabs of this nestlings species, in the Pantanal wetlands, in the state of Mato Grosso do Sul, Brazil. One Salmonella-like colony was serologically typed and identified as Salmonella Braenderup. Due to the high zoonotical potential of this microorganism, it is important an effective sanitary control of wildlife psittacines. In the literature searched it was not found any report on isolation of this bacterium in Hyacinth macaw for both free-living and captive animals.

Isolation of Arcobacter spp from the milk of dairy cows in Brazil

Bacteriologic examinations were performed on 188 milk samples collected from cows from 11 farms for diagnosis of mastitis in three cities of Rio Grande do Sul, Brazil. Among the common causes of mastitis, the most frequent isolates were Staphylococcus aureus, followed by Corynebacterium sp, Streptococcus uberis, Streptococcus dysgalactiae and Streptococcus agalactiae. Bacteriologic examination of 32 milk samples from one farm didn't show bacteria known as common etiologic agent of mastitis. Six samples of Arcobacter spp typed by genotypic tests as Arcobacter cryaerophilus (five strains) and Arcobacter butzleri (one strain) were isolated from cows' milk of that farm. It is reported the isolation of Arcobacter species from the milk of cows in absence of clinical signs of mastitis. This is the first report of the detection of the microorganisms in the milk of dairy cows in Brazil. No previous reports are known from other countries.

Foram realizados exames bacteriológicos em 188 amostras de leite colhidas de vacas de 11 propriedades leiteiras para diagnóstico de mastite, em três municípios no Rio Grande do Sul, Brasil. Entre as causas comuns de mastite, os isolados mais freqüentes foram Staphylococcus aureus, seguido de Corynebacterium sp, Streptococcus uberis, Streptococcus dysgalactiae e Streptococcus agalactiae. O exame bacteriológico realizado em 32 amostras de leite de vacas de uma propriedade não demonstrou a presença de bactérias conhecidas como causadoras de mastite. Foram isoladas do leite de vacas desta propriedade seis amostras de Arcobacter spp, classificadas por testes moleculares como Arcobacter cryaerophilus (cinco amostras) e Arcobacter butzleri (uma amostra). É relatado o isolamento de espécies de Arcobacter do leite de vacas na ausência de sinais clínicos de mastite. Este é o primeiro relato da detecção dos microorganismos no leite de vacas leiteiras no Brasil.

Diagnóstico molecular de Leptospira spp em matrizes suínas descartadas / Molecular diagnosis of Leptospira spp in culled sows

O diagnóstico da leptospirose foi efetuado através de método molecular, histopatológico e sorológico em 30 matrizes suínas, descartadas, no Rio Grande do Sul, Brasil. Os objetivos foram comparar a eficiência dos 3 métodos, verificar a sensibilidade de um método de PCR que utiliza um primer único baseado na seqüência de um elemento repetitivo do genoma de Leptospira interrogans, bem como verificar a possível detecção de leptospiras em vários tecidos , incluindo o trato genital. Os animais foram selecionados com base no teste de algutinação microscópica para incluir tanto animais negativos como positivos e com baixos e altos títulos sorológicos. As maiores freqüências (90% dos animais positivos) e títulos (100 a 800) foram observados para L. interrogans serovar bratislava. Leptospiras foram detectadas por histopatologia em apenas 9 matrizes, todas com altos títulos (pelo menos 100). Um produto de PCR de 438 bp foi observado em todos os animais (fragmentos de 25 rins, 24 úteros e 9 avidutos). Produtos de PCR similares foram obtidos em DNA de culturas de leptospiras patogênicas, enquanto a não patogênica, L. patoc apresentou um padrão distinto. Nenhum produto de amplificação de DNA de Leptospira spp foi detectado em DNA de culturas de Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella sp, Streptococcus sp and Staphylococus aureus, ou de sangue de dois leitões. O método molecular foi, assim , específico e o mais eficiente para detectar baixos níveis de patógeno, sendo capaz de difernciar leptospiras patogênicas e não patogênicas.

Leptospirosis diagnosis was performed through molecular, histopathological and serological tests in 30 culled sows in Rio Grande do Sul, Brazil. The objectives were to compare the efficiency of the three methods, to verify the sensitivity of a PCR methodology using a single primer based on the sequence of a repetitive element of Leptospira interrogans genome, as well as to verify the possible detection of Leptospira in several tissue including the genital tract of sows. The animals were selected based on the microscopic agglutination test in order to have sows with negative and positive results, presenting low and higher serologic titers. The higher frequency (90% of the positive sows) and titers (100 to 800) was observed for L. interrogans serovar bratislava. Leptospira was detected by histopathology in nine sows only, all presenting higher serologic titers (at least 100). A PCR product of 438 bp was observed in all animals (25 kidneys, 24 uterus and 9 oviduct) fragments. Similar PCR product was obtained for DNA from cultures of other pathogenic leptospires, while the pattern observed for the non-pathogenic L. patoc was distinct. No Leptospira spp DNA amplification product was detected in Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella sp, Streptococcus sp and Staphylococcus aureus DNAs obtained from cultures, or in blood DNA samples of two piglets. The molecular system was therefore specific and the most effective to detect low pathogen levels, being able to differentiate pathogenic from non-pathogenic leptospires.

Effects of irinotecan/5-fluorouracil combination on thymidylate synthase and cell cycle in human colon carcinoma cell line

Thymidylate synthase (TS) is responsible for the de novo synthesis of thymidylate,which is required for DNA synthesis and repair TS and is an important target for 5-fluorouracil (5-FU). In this study we investigated whether the greater cytotoxicity of irinotecan (CPT-11) followed by 5-FU in HT-29 and SNU-C4 cell lines was related to TSmRNA expression or activity. Thus, cells were exposed for 24 h to each drug, alone or in combination, and assessed for colony formation, TS catalytic activity, TS-mRNA expression and cell cycle distribution. Pre-treatment with CPT-11 at IC20 increased the 5-FU cytotoxicity in HT-29 and SNU-C4 cells. TS catalytic activity and TS-mRNA expression suggested that the differences in sensitivity to 5-FU among the cell lines were not correlated to TS profile. When we exposed cells to CPT-11 at IC20 and subsequently to 5-FU at IC50, the impact on TS activity and mRNA expression were the same as observed with 5-FU alone. CPT-11 induced G2/M arrest, while 5-FU arrested cells in S-phase in both cell lines. When cells were treated with CPT-11 followed by 5-FU we observed an increased in 5-FU-inducible S-phase arrest in both cell lines. Our findings suggested that the greater cytotoxicity of CPT-11/5-FU combination in HT-29 and SNU-C4 cell lines are not related to interference with thymidylate synthase.