Desempenho produtivo e reprodutivo de novilhas com lactação induzida e eficácia da terapia antimicrobiana associada com selante interno de tetos

This study evaluated (a) the efficacy of an association between injectable antibiotic therapy and sealant (ATBS) on milk yield (MY), somatic cell count (SCC), and prevalence of intramammary infections (IMI); and (b) the efficacy of gonadotropin-releasing hormone (GnRH) on follicular cyst (FCs) resolution (cyclicity at the 45th day in milk; DIM) and cumulative pregnancy rate (CPR) in heifers submitted to a lactation induction protocol (LIP). A total of 114 crossbred (Holstein × Jersey) heifers, with 34.7 ± 4.8 months and 439 ± 56.35 kg were submitted to LIP. On the 5th day of the LIP, the heifers were assigned to (i) ATBS (n = 57) with 7 mg/kg of norfloxacin associated with sealant and (ii) Control 1 (n = 57; CONT1) with no treatments. Lactation began on the 21st day of LIP and the 15th DIM, FCs were diagnosed and 106 heifers were randomized into two treatment groups with 53 heifers each: (i) GnRH (5 mL injectable GnRH) and (ii) Control 2 (CONT2; no treatment). Of the 114 heifers initially induced, 83.33% (n = 95) responded to LIP with an average MY of 15.19 kg/milk/day during 22 weeks of lactation. In the first 14 DIM, the IMI prevalence was 18% and 28% for heifers ATBS and CONT1 treated, respectively. Additionally, coagulase-negative Staphylococcus was the most frequently isolated group of pathogens. Mammary quarters that received ATBS treatment had a lower risk of IMI and SCC than CONT1. The cyclicity at 45 DIM was 68% (ATBS) and 35% (CONT1), and 57% and 46% for animals in the GnRH and CONT2. CPR was 60% in the ATBS group and 89% in CONT1, but GnRH treatment did not affect the CPR. In conclusion, LIP was effective in stimulating MY in heifers, and the IMI prevalence decreased with ATBS treatment. Also, the use of GnRH did not affect the FC regression, cyclicity at 45 DIM, and CPR.Este estudo avaliou a (i) eficácia da associação entre antibioticoterapia injetável e selante interno de tetos (ATBS) na produção de leite (PL), contagem de células somáticas (CCS), e prevalência de infecções intramamárias (IIM); e (ii) eficácia do hormônio liberador de gonadotrofina (GnRH) na resolução de cistos foliculares (CFs), ciclicidade ao 45º dia em lactação (DEL) e taxa de prenhez cumulativa (TPC) em novilhas submetidas a um protocolo de indução de lactação (PIL). Um total de 114 novilhas mestiças (Holandês × Jersey), com 34,7 ± 4,8 meses e 439 ± 56,35 kg foram submetidas ao PIL. No 5º dia do PIL, as novilhas receberam: (i) ATBS (n = 57) com 7 mg/kg de norfloxacina associada ao selante interno de tetos e (ii) Controle 1 (n = 57; CONT1) sem tratamento. A lactação teve início no 21º dia do PIL e no 15º DEL, foram diagnosticados CFs e 106 novilhas foram agrupadas em dois grupos de tratamento com 53 novilhas em cada: (i) GnRH (5 mL de GnRH injetável) e (ii) Controle 2 (CONT2; sem tratamento). Das 114 novilhas inicialmente induzidas, 83,33% (n = 95) responderam ao PIL com PL média de 15,19 kg/leite/d durante 22 semanas de lactação. Nos primeiros 14 DEL a prevalência de IIM foi de 18% e 28% para as novilhas tratadas com ATBS e CONT1, respectivamente. Além disso, estafilococos coagulase negativa foram o grupo de patógenos mais frequentemente isolados. Quartos mamários tratados com ATBS tiveram menor risco (0,56) de IIM e menor CCS do que CONT1. A ciclicidade a 45 DEL foi de 68% (ATBS) e 35% (CONT1), e 57% e 46% para os animais no GnRH e CONT2. A TPC foi de 60% no grupo ATBS e 89% no CONT1, porém o tratamento com GnRH não afetou a TPC. Em conclusão, o PIL foi eficaz em estimular a PL em novilhas tardias e a prevalência de IIM diminiuiu com o tratamento ATBS. Além disso, o uso de GnRH não afetou a regressão de CF, ciclicidade em 45 DEL e a TPC

Características seminais de ovinos suplementados ou não com uréia e diferentes fontes de enxofre

Twelve adult Santa Inês crossbred rams with similar weight and age employed in a randomized design for 60 days period to evaluate three treatments: A. 100% of degradable protein requirement (control); B. 100% of degradable protein requirement + 3% urea + inorganic sulphur (99%S) and C. 100% of degradable protein requirement +3% urea + organic sulphur (21,5% S). Every week seminal collections were made through artificial vagina; blood collections were made to analyze plasma N-ureic levels and measured live weight and scrotal circumference. In semen samples were studied volume, microscopic waves, vigor, motility, concentration, total sperm per ejaculate, total feasible sperm per ejaculate, membrane sperm integrity, acrosomal integrity, percent of abnormal spermatozoa and N-ureic level in seminal plasma. Treatments experimental animals receiving presented blood and seminal plasma N-ureic levels higher than the ones of control treatment (p < 0,05). There was significant difference between organic and inorganic sources of sulphur in the following semen characteristics (p < 0,05): treatment C presented microscopic waves (4,57), motility (85,69%), vigor (4,66) and total sperm per ejaculate (9,02 x 10(9)) higher than treatment B; and the percentage of secondary sperm abnormality (5,37%) was lower than treatment B.Doze carneiros machos adultos mestiços Santa Inês de mesma idade e porte semelhante foram empregados em um delineamento inteiramente casualizado, por um período experimental de 60 dias. Os animais foram distribuídos para três tratamentos: A. 100% das exigências em proteína degradável no rúmen (controle); B. 100% das exigências em proteína degradável no rúmen + 3% de uréia + enxofre (99% S) e C. 100% das exigências em proteína degradável no rúmen + 3% de uréia + enxofre quelatado (21,5% S). Semanalmente foram colhidas amostras de sêmen obtidas com emprego de vagina artificial e de sangue para determinação da concentração de nitrogênio uréico plasmático, assim como realizadas pesagens dos animais e aferições de circunferência escrotal. No sêmen foram analisados: volume e turbilhonamento; vigor, motilidade e concentração espermática; total de espermatozóides e total de espermatozóides viáveis no ejaculado; integridade de membrana e de acrossoma; morfologia espermática e concentração de nitrogênio uréico no plasma seminal. Os animais suplementados com uréia apresentaram níveis de N-uréico no plasma sanguíneo e seminal significativamente maiores que os encontrados nos do tratamentos controle (p<0,05). Houve diferença significativa entre as fontes de enxofre utilizadas (p<0,05) quanto às características do sêmen estudadas, o tratamento C apresentando valores maiores para turbilhonamento (4,57), motilidade espermática (85,69%), vigor espermático (4,66) e total de espermatozóides por ejaculado (9,02 x 10(9)), além de uma porcentagem inferior de defeitos menores (5,37%) quando comparado ao tratamento B

Reference gene selection for gene expression analysis of oocytes collected from dairy cattle and buffaloes during winter and summer

Oocytes from dairy cattle and buffaloes have severely compromised developmental competence during summer. While analysis of gene expression is a powerful technique for understanding the factors affecting developmental hindrance in oocytes, analysis by real-time reverse transcription PCR (RT-PCR) relies on the correct normalization by reference genes showing stable expression. Furthermore, several studies have found that genes commonly used as reference standards do not behave as expected depending on cell type and experimental design. Hence, it is recommended to evaluate expression stability of candidate reference genes for a specific experimental condition before employing them as internal controls. In acknowledgment of the importance of seasonal effects on oocyte gene expression, the aim of this study was to evaluate the stability of expression levels of ten well-known reference genes (ACTB, GAPDH, GUSB, HIST1H2AG, HPRT1, PPIA, RPL15, SDHA, TBP and YWHAZ) using oocytes collected from different categories of dairy cattle and buffaloes during winter and summer. A normalization factor was provided for cattle (RPL15, PPIA and GUSB) and buffaloes (YWHAZ, GUSB and GAPDH) based on the expression of the three most stable reference genes in each species. Normalization of non-reference target genes by these reference genes was shown to be considerably different from normalization by less stable reference genes, further highlighting the need for careful selection of internal controls. Therefore, due to the high variability of reference genes among experimental groups, we conclude that data normalized by internal controls can be misleading and should be compared to not normalized data or to data normalized by an external control in order to better interpret the biological relevance of gene expression analysis.The research was granted by São Paulo Research Foundation (FAPESP; grant numbers 2009/00938-3, 2010/13384-3, 2010/09561-7, 2011/14207-0 and 2012/07510-1), National Counsel of Technological and Scientific Development (CNPq; grant number 476229/2011-1) and Coordination for the Improvement of Higher Level Personnel (CAPES). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Características seminais de ovinos suplementados ou não com uréia e diferentes fontes de enxofre

Doze carneiros machos adultos mestiços Santa Inês de mesma idade e porte semelhante foram empregados em um delineamento inteiramente casualizado, por um período experimental de 60 dias. Os animais foram distribuídos para três tratamentos: A. 100% das exigências em proteína degradável no rúmen (controle); B. 100% das exigências em proteína degradável no rúmen + 3% de uréia + enxofre (99% S) e C. 100% das exigências em proteína degradável no rúmen + 3% de uréia + enxofre quelatado (21,5% S). Semanalmente foram colhidas amostras de sêmen obtidas com emprego de vagina artificial e de sangue para determinação da concentração de nitrogênio uréico plasmático, assim como realizadas pesagens dos animais e aferições de circunferência escrotal. No sêmen foram analisados: volume e turbilhonamento; vigor, motilidade e concentração espermática; total de espermatozóides e total de espermatozóides viáveis no ejaculado; integridade de membrana e de acrossoma; morfologia espermática e concentração de nitrogênio uréico no plasma seminal. Os animais suplementados com uréia apresentaram níveis de N-uréico no plasma sanguíneo e seminal significativamente maiores que os encontrados nos do tratamentos controle (p<0,05). Houve diferença significativa entre as fontes de enxofre utilizadas (p<0,05) quanto às características do sêmen estudadas, o tratamento C apresentando valores maiores para turbilhonamento (4,57), motilidade espermática (85,69%), vigor espermático (4,66) e total de espermatozóides por ejaculado (9,02 x 109), além de uma porcentagem inferior de defeitos menores (5,37%) quando comparado ao tratamento BTwelve adult Santa Inês crossbred rams with similar weight and age employed in a randomized design for 60 days period to evaluate three treatments: A. 100% of degradable protein requirement (control); B. 100% of degradable protein requirement + 3% urea + inorganic sulphur (99%S) and C. 100% of degradable protein requirement +3% urea + organic sulphur (21,5% S). Every week seminal collections were made through artificial vagina; blood collections were made to analyze plasma N-ureic levels and measured live weight and scrotal circumference. In semen samples were studied volume, microscopic waves, vigor, motility, concentration, total sperm per ejaculate, total feasible sperm per ejaculate, membrane sperm integrity, acrosomal integrity, percent of abnormal spermatozoa and N-ureic level in seminal plasma. Treatments experimental animals receiving presented blood and seminal plasma N-ureic levels higher than the ones of control treatment (p < 0,05). There was significant difference between organic and inorganic sources of sulphur in the following semen characteristics (p < 0,05): treatment C presented microscopic waves (4,57), motility (85,69%), vigor (4,66) and total sperm per ejaculate (9,02 x 109) higher than treatment B; and the percentage of secondary sperm abnormality (5,37%) was lower than treatment

Seminal characteristics of rams supplement or not with urea and two sources of sulphur

Doze carneiros machos adultos mestiços Santa Inês de mesma idade e porte semelhante foram empregados em um delineamento inteiramente casualizado, por um período experimental de 60 dias. Os animais foram distribuídos para três tratamentos: A. 100% das exigências em proteína degradável no rúmen (controle); B. 100% das exigências em proteína degradável no rúmen + 3% de uréia + enxofre (99% S) e C. 100% das exigências em proteína degradável no rúmen + 3% de uréia + enxofre quelatado (21,5% S). Semanalmente foram colhidas amostras de sêmen obtidas com emprego de vagina artificial e de sangue para determinação da concentração de nitrogênio uréico plasmático, assim como realizadas pesagens dos animais e aferições de circunferência escrotal. No sêmen foram analisados: volume e turbilhonamento; vigor, motilidade e concentração espermática; total de espermatozóides e total de espermatozóides viáveis no ejaculado; integridade de membrana e de acrossoma; morfologia espermática e concentração de nitrogênio uréico no plasma seminal. Os animais suplementados com uréia apresentaram níveis de N-uréico no plasma sanguíneo e seminal significativamente maiores que os encontrados nos do tratamentos controle (p<0,05). Houve diferença significativa entre as fontes de enxofre utilizadas (p<0,05) quanto às características do sêmen estudadas, o tratamento C apresentando valores maiores para turbilhonamento (4,57), motilidade espermática (85,69%), vigor espermático (4,66) e total de espermatozóides por ejaculado (9,02 x 109), além de uma porcentagem inferior de defeitos menores (5,37%) quando comparado ao tratamento BTwelve adult Santa Inês crossbred rams with similar weight and age employed in a randomized design for 60 days period to evaluate three treatments: A. 100% of degradable protein requirement (control); B. 100% of degradable protein requirement + 3% urea + inorganic sulphur (99%S) and C. 100% of degradable protein requirement +3% urea + organic sulphur (21,5% S). Every week seminal collections were made through artificial vagina; blood collections were made to analyze plasma N-ureic levels and measured live weight and scrotal circumference. In semen samples were studied volume, microscopic waves, vigor, motility, concentration, total sperm per ejaculate, total feasible sperm per ejaculate, membrane sperm integrity, acrosomal integrity, percent of abnormal spermatozoa and N-ureic level in seminal plasma. Treatments experimental animals receiving presented blood and seminal plasma N-ureic levels higher than the ones of control treatment (p < 0,05). There was significant difference between organic and inorganic sources of sulphur in the following semen characteristics (p < 0,05): treatment C presented microscopic waves (4,57), motility (85,69%), vigor (4,66) and total sperm per ejaculate (9,02 x 109) higher than treatment B; and the percentage of secondary sperm abnormality (5,37%) was lower than treatment

Progestin-Based Protocol for Synchronization of Follicular Wave Emergence in Buffaloes During Summer and Winter

The aim of the present study was to evaluate the effects of type of norgestomet auricular implant (new - N or previously used during 5 days - U), season of the year (summer - S and winter - W), and parity (12 heifers - H and 23 cows - C) on synchronization of follicular wave emergence in buffaloes. For this purpose, 35 buffaloes were examined daily by ultrasonography until follicular wave emergence was detected. Data were analysed by ANOVA, using PROC GLIMMIX. No interactions were observed in none variables. Time of follicular wave emergence and number of follicles at emergence were not affected by type of implant or season of the year. Parity also did not influence the number of follicles at emergence. However, follicular wave emergence occurred later in heifers than in cows. In conclusion, the previous use of a norgestomet auricular implant independent of the season of the year does not affect the time or the number of follicles at follicular wave emergence in buffaloes. Nevertheless, although heifers and cows had a similar number of follicles at emergence, the time of follicular wave emergence occurs earlier in cows than in heifers

Comparação das técnicas de ELISA indireto e de soroneutralização na detecção de anticorpos contra o BHV-1 em amostras de soro bubalino (Bubalus bubalis)

O desempenho de um ELISA comercial (Bovine Rhinotracheitis virus antibody test Kit, Herdcheck®, IDEXX Laboratories Inc., USA) para a identificação de anticorpos anti-herpesvírus bovino tipo 1 (BHV-1), em soro bovino, foi avaliado em 133 amostras de soro de búfalos (Bubalus bubalis), sem histórico de vacinação para IBR, da Região do Vale do Ribeira, Estado de São Paulo. Tomando-se a técnica de soroneutralização como referência, o ELISA indireto apresentou sensibilidade e especificidade relativa de 97,14 e 46,03%, respectivamente. A concordância estimada pelo índice kappa, entre os dois métodos, foi de 0,44 IC (0,29-0,59), utilizando um intervalo de confiança de 95%

Xenooplasmic Transfer between Buffalo and Bovine Enables Development of Homoplasmic Offspring

Nuclear-mitochondrial incompatibilities may be responsible for the development failure reported in embryos and fetuses produced by interspecies somatic cell nuclear transfer (iSCNT). Herein we performed xenooplasmic transfer (XOT) by introducing 10 to 15% of buffalo ooplasm into bovine zygotes to assess its effect on the persistence of buffalo mitochondrial DNA (mtDNA). Blastocyst rates were not compromised by XOT in comparison to both in vitro fertilized embryos and embryos produced by transfer of bovine ooplasm into bovine zygotes. Moreover, offspring were born after transfer of XOT embryos to recipient cows. Buffalo mtDNA introduced in zygotes was still present at the blastocyst stage (8.3 vs. 9.3%, p = 0.11), indicating unaltered heteroplasmy during early development. Nonetheless, no vestige of buffalo mtDNA was found in offspring, indicating a drift to homoplasmy during later stages of development. In conclusion, we show that the buffalo mtDNA introduced by XOT into a bovine zygote do not compromise embryo development. On the other hand, buffalo mtDNA was not inherited by offspring indicating a possible failure in the process of interspecies mtDNA replication.FAPESP, Sao Paulo, SP, Brazil[2002/05054-7]FAPESP, Sao Paulo, SP, Brazil[2004/01841-0]FAPESP, Sao Paulo, SP, Brazil[2006/59074-0