Shiga toxigenic and enteropathogenic Escherichia coli in water and fish from pay-to-fish ponds.

UNLABELLED: Escherichia coli is part of the normal microflora of the intestines of mammals. However, among the enteric pathogens, it is one of the leading causes of intestinal diseases, especially Shiga toxigenic E. coli, which can cause diarrhoea, haemorrhagic colitis and complications like haemolytic uraemic syndrome and thrombotic thrombocytopaenic purpura. Escherichia coli is considered a serious public health problem. Water and fish samples were subjected to biochemical tests to confirm the presence of E. coli and by PCR to verify the presence of pathogenic strains (O157, enteropathogenic and shiga toxigenic) in water and fish (skin, gastrointestinal tract and muscles) from pay-to-fish ponds located in the Córrego Rico watershed in the northeastern region of the state of São Paulo, Brazil. Of the 115 E. coli isolates from fish or water, five (4·34%) contained eae and stx2 genes, one had only the eae gene and two had the stx1 gene. An isolate containing the stx2 gene was also found in the water sample. In addition, eight isolates (6·95%) from the fish gastrointestinal tract contained rfbEO157:H7 (O157 gene), and three (2·61%) contained stx2 and eae genes, demonstrating the potential risk to the environment and public health. The results provide useful basic information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population. SIGNIFICANCE AND IMPACT OF THE STUDY: Pay-to-fish ponds are a common commercial activity in Brazil. Samples of water and Oreochromis niloticus were examined by PCR to detect the presence of pathogenic strains of Escherichia coli (O157, enteropathogenic and shiga toxigenic). Several pathogenic strains were detected in this study, providing useful epidemiological information for the proper management of these environments and animals in order to prevent faecal pollution, reducing health risks to the Brazilian population.

Corpus luteum blood flow evaluation on Day 21 to improve the management of embryo recipient herds.

The aim of the present study was to use blood flow evaluation of the CL at 14 days after embryo transfer to detect nonpregnant animals and optimize the management of bovine recipients. The estrous cycle was synchronized in 165 recipients, and the day of expected ovulation was considered to be Day 0. Embryo transfer was performed 7 days later, on Day 7. On Day 21, pregnancy was diagnosed on the basis of blood flow evaluation of the CL (DG21-predictive diagnostic). To validate this methodology, visual scores for blood flow were compared to objective data extracted from CL ultrasound images recorded in the Doppler mode. The size was also evaluated using recorded images of the CL in the B mode. Blood samples were also collected for further analysis of the progesterone (P4) concentration. The diagnosis of pregnancy was confirmed at 35 days after estrus (DG35-definitive diagnostic). The DG21 showed that 55.2% (90 of 163) of the animals were presumptively pregnant, and this value was higher (P < 0.04) than that obtained at DG35 (43.6%, 71 of 163). The predictive diagnostic achieved moderate specificity (79.3%) for the detection of pregnancy, but most importantly, high sensitivity (100%) for the detection of nonpregnant recipients. The overall accuracy of the diagnosis was 88.3%. The P4 concentrations were different (P < 0.02) and correlated with each visual score assigned for the CL size. Visual scores for CL blood flow were also efficient (P < 0.0001) to distinguish animals with different levels of P4; however, P4 concentrations were higher for scores 1 and 2 (high and regular blood flow, respectively) than those for score 3 (low blood flow). This technique showed high sensitivity and facilitated the early detection of nonpregnant animals. The DG21 would allow about 79.3% of nonpregnant animals to be resynchronized 9 to 14 days earlier, when compared to conventional management based on pregnancy diagnosis at Days 30 to 35.

Perfil de progesterona em novilhas com diferente atividade luteal e tratadas com implantes vaginais / Serum progesterone in heifers with different luteal activity treated with vaginal devices

O dispositivo de liberação de progesterona (DLP) é muito importante em protocolos de Inseminação Artificial em Tempo Fixo (IATF). Representa cerca de 43% dos custos e é objeto de estudos sobre a eficiência da sua reutilização. No entanto, perfis de liberação de progesterona (P4) em animais com diferentes concentrações endógenas desse esteroide não são claramente descritos. Este estudo teve como objetivo avaliar a concentração sérica de P4 em fêmeas com diferentes situações de atividade luteal, tratadas com DLP novo (1g de P4) por 8 dias. Trinta novilhas mestiças cíclicas foram divididas em três grupos: em G1 e G2, o DLP foi inserido (D0) sete dias após a ovulação induzida. Adicionalmente, 0,15mg de D-cloprostenol foi administrado três dias depois para promover a luteólise em G2. Para G3, o corpo lúteo foi lisado antes da inserção do DLP para que a P4 exógena fosse a única fonte desse hormônio. O sangue foi coletado no D0, D3, D5 e D8, e a P4 avaliada por RIA. Médias de P4 foram comparadas entre os grupos em cada dia e dentro do grupo, entre os dias, utilizando o teste Tukey. Antes da inserção do implante (D0), os níveis de P4 foram, nos grupos, semelhantes em G1 e G2, e superiores a G3 (5,3±3,1a e 5,3±1,4avs 0,6±0.3bng/mL, respectivamente-P<0,05). No D3, ocorreu o mesmo perfil (5,7±2,6a e 5,4±2,0a e 3,6±0.8bng/mL, respectivamente para G1 e G2 vs G3, P<0,05). Trinta e seis horas (D5) após a PGF, a P4 no G2 caiu para níveis semelhantes aos do grupo G3 e ambos diferiram (P<0,05) de G1 (3,3±1,6b vs 2,4±0,9b e 2,1±0.7bng/mL). Essa diferença se manteve (P<0,05) em D8 (3,1±1,3a, 1,8±0,8b e 1,6±0.6b ng/mL). O aumento da P4 após a inserção (D3 - D0) foi maior (P<0,05) em G3 que em G1 e G2 (2,8±0,9a vs 0,4±1,8b e 0,2±1.4bng/mL). Os animais com maior P4 endógena levam a menor liberação de P4 exógena a partir do DLP. Portanto, os níveis remanescentes de P4 no DLP após o uso dependem da concentração endógena de P4 do animal e possíveis alterações durante a permanência.(AU)

The progesterone (P4) device is a very important step in the ovulation control in Timed Artificial Insemination (TAI) protocols. It represents about 43% of the hormone costs, thus it has been the subject of several studies on efficiency of the reused device as an alternative to reduce TAI costs. However, to our knowledge, profiles for P4 release in animals with different endogenous concentrations of P4 are not clearly described. This study aimed to evaluate serum concentration of P4 in females with different ovarian conditions - related to luteal activity - and treated with a new intravaginal device (1g of P4) for 8 days. Thirty normally cyclic crossbred heifers were divided into three groups: for G1 and G2, P4 device was inserted (D0) seven days after ovulation (7 day old CL). Additional PGF (0.15 mg of D-cloprostenol) was given three days later to promote luteolysis in the G2 group. For G3, CL was killed before P4 insertion and the exogenous progesterone was the only source of this hormone. Blood samples were collected on D0, D3, D5 and D8 and P4 concentration was measured by radioimmunoassay (RIA). Means for P4 concentration were compared among groups in each day and within the group among days using the Tukey test. Before P4 device insertion (D0), P4 levels were higher (P<0.05) in G1 and G2 when compared to G3 (5.3±3.1 and 5.3±1.4 vs. 0.6±0.3ng/mL, respectively). Three days later (D3), the same pattern was observed (5.7±2.6 and 5.4±2.0 and 3.6±0.8ng/mL, respectively for G1 and G2 vs. G3, P<0.05). Thirty-six hours (D5) after PGF injection (G2), P4 in G2 dropped to levels similar to the G3 group and both differed (P<0.05) from G1 (3.3±1.6 vs. 2.4±0.9 and 2.1±0.7ng/mL, G1 vs. G2 and G3, respectively). There were no differences (P>0.05) among groups on D8 (3.1±1.3, 1.8±0.8 and 1.6±0.6ng/mL, respectively, for G1, G2 and G3). Progesterone increase after P4 insertion (D3 - D0) was higher (P<0.05) in G3 compared to G1 and G2 (2.8±0.9 vs. 0.4±1.8 and 0.2±1.4ng/mL, respectively). The interpretation was that animals with higher endogenous P4 promote less release of the exogenous P4 from the device. Therefore, the remaining P4 levels from used progesterone devices depend on the physiological condition of the animal at the time of insertion and possible changes during the treatment.(AU)

Chicken skeletal muscle-associated macroarray for gene discovery.

Macro- and microarrays are well-established technologies to determine gene functions through repeated measurements of transcript abundance. We constructed a chicken skeletal muscle-associated array based on a muscle-specific EST database, which was used to generate a tissue expression dataset of ~4500 chicken genes across 5 adult tissues (skeletal muscle, heart, liver, brain, and skin). Only a small number of ESTs were sufficiently well characterized by BLAST searches to determine their probable cellular functions. Evidence of a particular tissue-characteristic expression can be considered an indication that the transcript is likely to be functionally significant. The skeletal muscle macroarray platform was first used to search for evidence of tissue-specific expression, focusing on the biological function of genes/transcripts, since gene expression profiles generated across tissues were found to be reliable and consistent. Hierarchical clustering analysis revealed consistent clustering among genes assigned to 'developmental growth', such as the ontology genes and germ layers. Accuracy of the expression data was supported by comparing information from known transcripts and tissue from which the transcript was derived with macroarray data. Hybridization assays resulted in consistent tissue expression profile, which will be useful to dissect tissue-regulatory networks and to predict functions of novel genes identified after extensive sequencing of the genomes of model organisms. Screening our skeletal-muscle platform using 5 chicken adult tissues allowed us identifying 43 'tissue-specific' transcripts, and 112 co-expressed uncharacterized transcripts with 62 putative motifs. This platform also represents an important tool for functional investigation of novel genes; to determine expression pattern according to developmental stages; to evaluate differences in muscular growth potential between chicken lines, and to identify tissue-specific genes.

Role of the oblique ligament in the integrity of the medial collateral ligament of the canine elbow joint

It was studied the arrangement of the collagen fibrils of the medial collateral ligament of the canine elbow joint and evaluated its diameter, when it was isolated or associated to the oblique ligament and loaded in tension until failure. Eighteen joints were divided in three groups. The first group had the medial collateral ligament collected and not loaded, the second group had the medial collateral ligament tested separately and the third group had both ligaments associately tested. Medial collateral ligament not submitted to strain presented a wavy and reticular pattern of the collagen fibers, which was not totally destroyed when it was loaded associated to the oblique ligament, and totally loses the reticular pattern when stretched separately. When the medial collateral ligament was loaded in tension separately, the mean collagen fibrils diameter increased in relation to the group not submitted to the tensile strain. Associated to the oblique ligament, the mean collagen fibrils diameter was the largest in the insertion area and the smallest in the mid-substance, in relation to the other groups. It was concluded that the oblique ligament could favor the integrity of the medial collateral ligament insertion area, facilitating its reconstruction after lesion with larger efficiency.

Foram estudados o arranjo e o diâmetro médio das fibrilas colágenas do ligamento colateral medial da articulação do cotovelo do cão, isolado ou associado ao ligamento oblíquo e tracionado até a ruptura. Dezoito articulações foram divididas em três grupos. O primeiro grupo teve o ligamento colateral medial coletado, mas não tracionado; o segundo grupo teve o ligamento colateral medial tracionado isoladamente; o terceiro grupo teve os ligamentos colateral medial e oblíquo tracionados associadamente. O ligamento colateral medial não submetido ao ensaio de tração apresentou um padrão ondulado das fibras colágenas, o qual não foi totalmente destruído quando foi tracionado, associado ao ligamento oblíquo, e perdeu totalmente o padrão reticular das fibras colágenas quando testado isoladamente. Quando o ligamento colateral medial foi submetido à tensão isoladamente, o diâmetro médio das fibrilas colágenas aumentou em relação ao grupo não submetido à tensão. Associado ao ligamento oblíquo, o diâmetro médio das fibrilas colágenas foi o maior na região de inserção e o menor na região média, em relação aos outros grupos. Concluiu-se que o ligamento oblíquo pode favorecer a integridade da região de inserção do ligamento colateral medial, aumentando a eficácia de sua reconstrução após a lesão.