Embrapa Dairy Cattle Research Center

Juiz de Fora, Brazil

Embrapa Dairy Cattle Research Center

Juiz de Fora, Brazil
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Viana J.H.M.,Embrapa Dairy Cattle Research Center | Palhao M.P.,Federal University of Viçosa | Siqueira L.G.B.,Embrapa Cerrados | Fonseca J.F.,Embrapa Goats and Sheep Research Center | Camargo L.S.A.,Embrapa Dairy Cattle Research Center
Theriogenology | Year: 2010

The objective of this study was to evaluate ovarian follicular dynamics during intervals between successive ovum pick-up (OPU) and determine its effects on the number and quality of recovered cumulus-oocyte complexes (COCs) in Zebu cows (Bos indicus). Pluriparous nonlactating Gyr cows (Bos indicus; n = 10) underwent four consecutive OPU sessions at 96-h intervals. The dynamics of ovarian follicular growth between OPU sessions was monitored by twice-daily ultrasonographic examinations. A single dominant follicle (DF) or two codominant (CDF) follicles (>9 mm) were present in 63.3% (19 of 30) of intervals studied, with follicle deviation beginning when the future dominant follicle (F1) achieved a diameter of 6.2 ± 0.3 mm. The phenomenon of codominance was observed in four (13.3%) of the inter-OPU intervals. The remaining intervals (36.6%, 11 of 30) were characterized by a greater follicular population, lower rate of follicular growth, and a smaller diameter F1 (P < 0.0001). There was a tendency (P = 0.08) toward an increase in the number of recovered COCs when dominant follicles were not present (NDF). The quality of COCs was not affected by the presence of a single dominant follicle, but codominant follicles resulted in recovery of a lower proportion of viable embryos (40.0%, 62.1%, and 63.6%; P < 0.05) and higher proportions of degenerate COCs (56.0%, 30.3%, and 28.6%; P < 0.05) for CDF, NDF, and DF respectively. We concluded that, in Zebu cows, (a) repeated follicle aspirations altered ovarian follicular dynamics, perhaps by increasing follicular growth rate; (b) follicular dominance could be established in cows undergoing twice-a-week OPU; and (c) the presence of a dominant follicle during short inter-OPU intervals may not affect COC quality, except when a codominant follicle was present. © 2010 Elsevier Inc. All rights reserved.

Souza J.M.G.,Federal University of Viçosa | Torres C.A.A.,Federal University of Viçosa | Maia A.L.R.S.,Federal University of Fluminense | Brandao F.Z.,Federal University of Fluminense | And 4 more authors.
Animal Reproduction Science | Year: 2011

Intravaginal progesterone devices are used worldwide for estrus induction in goats. Reused devices are able to induce estrus; however, this can be a health risk within a flock. The objective was to compare new and previously used (and autoclaved) progesterone-releasing intravaginal devices for induction of estrus and ovulation in seasonally anestrous Toggenburg goats. Anestrous goats (n=42) received new intravaginal devices containing 0.3. g progesterone (CONTROL), or similar devices previously used for either 6 (USED6) or 12. d (USED12) and subsequently autoclaved. All goats received 5. mg dinoprost at device insertion and 200 IU eCG 5. d later, and all devices were removed after 6. d. After device removal, estrus was monitored and females displaying signs of estrus were mated by fertile bucks. Transrectal ovarian ultrasonography was performed after device removal until detection of ovulation. Blood samples were collected for determination of plasma progesterone concentration at different times. There was no difference (P>0.05) among groups CONTROL, USED6 or USED12 for: estrus response (87, 100 or 100%, respectively); duration of estrus (32.3±2.3, 25.2±3.4 or 27.3±4.1. h); ovulation rate (100, 88 or 100%); number of ovulations (1.5±0.2, 1.9±0.3 or 1.7±0.3); and pregnancy rate (60, 58 or 67%). Plasma progesterone (P4) concentrations were greater (P<0.05) in CONTROL than in USED6-treated and USED12-treated goats (7.2±1.2, 4.7±0.7 and 4.3±0.6. ng/mL, respectively) at 6. h after device insertion; these differences were maintained until 4. d after device insertion (3.4±0.4, 2.3±0.2, and 2.5±0.2. ng/mL). Overall, plasma progesterone concentrations were greater (P<0.05) in nulliparous than in lactating goats (3.1±0.8 compared to 2.4±0.6. ng/mL, respectively). In conclusion, autoclaved, previously used intravaginal progesterone-releasing devices resulted in significant lesser plasma progesterone concentrations than new devices, but were similarly effective in inducing estrus and ovulation in anestrous goats. © 2011 Elsevier B.V.

Azevedo A.L.S.,Embrapa Dairy Cattle Research Center | Costa P.P.,Embrapa Dairy Cattle Research Center | Machado J.C.,Embrapa Dairy Cattle Research Center | Machado M.A.,Embrapa Dairy Cattle Research Center | And 2 more authors.
Crop Science | Year: 2012

Napier grass (Pennisetum purpureum Schum.) is an important forage crop in tropical areas although little is known about its genome information, and few molecular markers have been developed for this species. This work aimed to check the viability of cross-species amplification of microsatellite markers between pearl millet (Pennisetum glaucum) and Napier grass and to evaluate the genetic diversity among Napier grass germplasm accessions. Fifty-four microsatellite markers previously described in pearl millet were tested against Napier grass, and 30 markers (55.5%) showed successful cross-amplification. From them, 18 microsatellite markers were selected to study the genetic diversity in the Embrapa Active Germplasm Bank of Napier Grass (Embrapa-BAGCE). A total of 180 alleles were identified by these selected microsatellite markers in 107 Napier grass accessions and four pearl millet samples. The average similarity coefficient (Dice) calculated among the Embrapa-BAGCE accessions was 0.651, ranging from 0.254 to 1.0. Some accessions showed similarity coefficients equal to one, indicating that they have common progenitors or that they might be the same accessions with different denomination. To our knowledge, this work is the first to describe microsatellite markers in Napier grass and represents a significant advance regarding the use of molecular markers in this species. © Crop Science Society of America.

Oliveira C.S.,São Paulo State University | Oliveira C.S.,Embrapa Dairy Cattle Research Center | Saraiva N.Z.,São Paulo State University | Cruz M.H.C.,São Paulo State University | And 4 more authors.
Reproduction | Year: 2013

During initial development, both X chromosomes are active in females, and one of them must be silenced at the appropriate time in order to dosage compensate their gene expression levels to male counterparts. Silencing involves epigenetic mechanisms, including histone deacetylation. Major X chromosome inactivation (XCI) in bovine occurs between hatching and implantation, although in vitro culture conditions might disrupt the silencing process, increasing or decreasing X-linked gene expression. In this study, we aimed to address the roles of histone deacetylase inhibition by trichostatin A (TSA) on female preimplantation development.We tested the hypothesis that by enhancing histone acetylation, TSA would increase the percentage of embryos achieving 16-cell stage, reducing percentage of embryos blocked at 8-cell stage, and interfere with XCI in IVF embryos. We noticed that after TSA treatment, acetylation levels in individual blastomeres of 8-16 cell embryos were increased twofold on treated embryos, and the samewas detected for blastocysts. Changes among blastomere levels within the same embryo were diminished on TSA group, as low-acetylated blastomeres were no longer detected. The percentage of embryos that reached the 5th cleavage cycle 118 h after IVF, analyzed by Hoechst staining, remained unaltered after TSA treatment. Then, we assessed XIST and G6PD expression in individual female bovine blastocysts by quantitative real-time PCR. Even though G6PD expression remained unaltered after TSA exposure, XIST expression was eightfold decreased, and we also detected a major decrease in the percentage of blastocysts expressing detectable XIST levels after TSA treatment. Based on these results, we conclude that HDAC is involved on XCI process in bovine embryos, and its inhibition might delay X chromosome silencing and attenuate aberrant XIST expression described for IVF embryos. © 2013 Society for Reproduction and Fertility.

Resende T.T.,Embrapa Dairy Cattle Research Center | Auad A.M.,Embrapa Dairy Cattle Research Center | Fonseca M.G.,Embrapa Dairy Cattle Research Center
Journal of Economic Entomology | Year: 2014

This study determined the number of spittlebug adults, Mahanarva spectabilis Distant (Hemiptera: Cercopidae), that should be used in selection tests of the forage grass, Brachiaria ruziziensis (Germain and Evrard). In this study, 0, 1, 2, 4, or 8 M. spectabilis adults were kept in plants for 4 or 8 d per experimental plot. After these periods, the insects were removed from the plants and chlorophyll content, damage score, dry weight, fresh weight, and percent dry matter of shoots were evaluated. Chlorophyll content decreased significantly with higher density of M. spectabilis in plants exposed to the pest for 4 or 8 d. Plants that were exposed to eight spittlebugs for 8 d showed a ≈60% loss of chlorophyll content. When the forage was infested with eight adults for 4 d, the average damage score was 3 (50% of the leaf area was affected). The damage score and fresh and dry weights of the forage did not change depending on the exposure time of the plants to the spittlebugs. The percentage of dry matter of the plants infested was higher with the increase insect density and exposure time for all densities. Thus, the minimum recommended number is eight M. spectabilis adults for 4 d in resistance tests of B. ruziziensis to this pest species. © 2014 Entomological Society of America.

Auad A.M.,Embrapa Dairy Cattle Research Center | Resende T.T.,Embrapa Dairy Cattle Research Center | da Silva D.M.,Federal University of Lavras | das Gracas Fonseca M.,Embrapa Dairy Cattle Research Center
Agroforestry Systems | Year: 2012

The objective of this work was to estimate the abundance, diversity and constancy of families from the order Hymenoptera, such as the seasonality of those through a survey on the insect population in a silvopastoral system. We installed a Malaise-type trap in a Brachiaria decumbens area managed by a silvopastoral system in Coronel Pacheco, MG, from August 2006 to July 2008. The trapped insects were screened, and those of the order Hymenoptera were classified into their respective family categories and quantified. We adopted the methodology of Bodenheimer to calculate the indices of constancy, while other indices were estimated PAST program. We sampled 5841 specimens in total, which included 549 morphospecies and were distributed among 11 families. Of the total specimens sampled, 80% were Formicidae, which besides being the most abundant, was also the most diverse and constant family. When entomophagous insects were analyzed, the highest values for these indices were recorded for the families Ichneumonidae, Braconidae, and Vespidae. Pollinators were less prevalent compared to the total number of sampled individuals. The population density in the Hymenoptera was not correlated between the two sampling years and climate factors. © 2011 Springer Science+Business Media B.V.

Auad A.M.,Embrapa Dairy Cattle Research Center | Carvalho C.A.,Embrapa Dairy Cattle Research Center | Clemente M.A.,Federal University of Juiz de fora | Prezoto F.,Federal University of Juiz de fora
Sociobiology | Year: 2010

Despite the important action of wasps, there are yet no published studies of the species makeup of this group in silvipastoral systems. Therefore, the aim of this work was to carry out an inventory of the social wasp fauna in this system, to clarify the abundance, richness and diversity of the group and to evaluate the constancy and seasonality of the species present. The study was carried out at the experimental field station of the Embrapa Dairy Cattle Research Center, in Southeast Brazil, during the period from July 2006 to June 2008. A malaise trap was used to collect the insects. We captured a total of 205 social wasp specimens, distributed in 13 morpho-species belonging to 4 genera. The genus Agelaia was most abundant, represented by the species Agelaia multipicta and Agelaia vicina. The diversity index in the present study was H' = 1.642 and the richness was S' = 13 species. There were more individuals captured during the rainy season than in the dry season. The richness and diversity indices were also higher in the rainy season.

Rodrigues M.F.,Federal University of Juiz de fora | Alves C.C.S.,Federal University of Juiz de fora | Figueiredo B.B.M.,Federal University of Juiz de fora | Rezende A.B.,Federal University of Juiz de fora | And 4 more authors.
Immunology | Year: 2013

Apoptosis of macrophages has been reported as an effective host strategy to control the growth of intracellular pathogens, including pathogenic mycobacteria. Tumour necrosis factor-α (TNF-α) plays an important role in the modulation of apoptosis of infected macrophages. It exerts its biological activities via two distinct cell surface receptors, TNFR1 and TNFR2, whose extracellular domain can be released by proteolysis forming soluble TNF receptors (sTNFR1 and sTNFR2). The signalling through TNFR1 initiates the majority of the biological functions of TNF-α, leading to either cell death or survival whereas TNFR2 mediates primarily survival signals. Here, the expression of TNF-α receptors and the apoptosis of alveolar macrophages were investigated during the early phase of infection with attenuated and virulent mycobacteria in mice. A significant increase of apoptosis and high expression of TNFR1 were observed in alveolar macrophages at 3 and 7 days after infection with attenuated Mycobacterium bovis but only on day 7 in infection with the virulent M. bovis. Low surface expression of TNFR1 and increased levels of sTNFR1 on day 3 after infection by the virulent strain were associated with reduced rates of apoptotic macrophages. In addition, a significant reduction in apoptosis of alveolar macrophages was observed in TNFR1-/- mice at day 3 after bacillus Calmette-Guérin infection. These results suggest a potential role for TNFR1 in mycobacteria-induced alveolar macrophage apoptosis in vivo. In this scenario, shedding of TNFR1 seems to contribute to the modulation of macrophage apoptosis in a strain-dependent manner. © 2013 John Wiley & Sons Ltd.

PubMed | Instituto Nacional de Tecnologia Agropecuaria, Federal University of Lavras and Embrapa Dairy Cattle Research Center
Type: Journal Article | Journal: Veterinary research communications | Year: 2016

Staphylococcus aureus is a pathogen that frequently causes mastitis in bovine herds worldwide. This pathogen produces several virulence factors, including cell-associated adhesins, toxic and cytolytic exoproteins, and capsular polysaccharides. The aim of the present study was to test for the presence of genes involved in capsular polysaccharide production and biofilm formation in S. aureus isolated from bovine mastitis samples collected from 119 dairy herds located in three different Brazilian regions, as well as to assay the production of capsular polysaccharides and biofilm, in vitro. The detection of the cap, icaAD, and bap genes was performed using PCR. The detection and quantification of capsular polysaccharide production was performed using ELISA assays. The ability of the isolates to form a biofilm was examined using the polystyrene surface of microtiter plates. All 159S. aureus isolates investigated harboured the cap gene: 80% carried the cap5 gene and 20% carried the cap8 gene. Sixty-nine percent of the isolates expressed capsular polysaccharide (CP) in vitro, 58% expressed CP5 and 11% expressed CP8. All of the isolates harboured the icaA and icaD genes, and 95.6% of the isolates carried the bap gene. Of the 159 isolates analysed, 97.5% were biofilm producers. A significant association between the capsular genotype and phenotype and the amount of biofilm formation was detected: cap5/CP5 isolates tended to form more biofilm and to produce a thinner CP layer than cap8/CP8 isolates. The results indicate a high potential for pathogenicity among S. aureus isolated from bovine milk collected from three different regions in Brazil.

PubMed | Federal University of Juiz de fora and Embrapa Dairy Cattle Research Center
Type: Journal Article | Journal: Journal of dairy science | Year: 2016

Bacteria of the genus Staphylococcus are one of the major pathogens causing bovine mastitis. In recent decades, resistance of this genus to oxacillin (methicillin) has been a matter of concern due to the possibility of reducing the effectiveness of mastitis treatments and the transfer of resistance determinants to other bacteria. Oxacillin resistance was studied in 170 staphylococci from bovine milk samples, including 79 Staphylococcus aureus and 91 coagulase-negative staphylococci (CNS). The susceptibility profile of 10 antimicrobial agents used in veterinary practice was determined by the Etest method. In addition to the Etest, the phenotypic characterization of oxacillin resistance was tested using the cefoxitin disk diffusion test. All isolates were screened by PCR to detect the mecA gene in 2 different regions of the gene. The isolates with an oxacillin minimum inhibitory concentration 0.5 g/mL or resistant to cefoxitin were identified by sequencing a 536-bp fragment of the 16S rRNA gene. This group of isolates was also evaluated for the presence of blaZ and mecC genes. Molecular analysis of the mecA gene was carried out by typing of the staphylococcal cassette chromosome mec (SCCmec). The relatedness of the mecA-positive isolates was evaluated by macrorestriction of chromosomal DNA followed by pulsed-field gel electrophoresis. With the exception of penicillin and oxacillin, 86% of the isolates showed susceptibility to cephalothin, gentamicin, erythromycin, sulfonamide, trimethoprim-sulfamethoxazole, and tetracycline. All S. aureus isolates were susceptible to oxacillin, whereas 47% (n=43) of the CNS isolates were resistant. The CNS isolates showed a higher resistance to cephalothin, erythromycin, tetracycline, and gentamicin in comparison with S. aureus. The mecA gene was only detected in 10 CNS isolates, identified as Staphylococcus epidermidis, and classified into 3 pulsotypes (A, B, and C) and 4 subtypes (A1, B1, B2, and B3). Among the isolates with an oxacillin resistance phenotype, 12 were positive for the blaZ gene, and 9 of them were mecA-positive. Two of the oxacillin-resistant isolates amplified the mecA homolog gene of Staphylococcus sciuri and none amplified mecC. Three SCCmec types, I, IV, and V, were found. Our results suggest that Staphylococcus epidermidis can be a reservoir for mecA for other Staphylococcus species. Studies investigating the molecular and phenotypic profile of antimicrobial resistance in staphylococcal species should be performed for controlling the spread of resistance and the selection of appropriate therapeutic measures.

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