Sperm membrane integrity and stability after selection of cryopreserved ovine semen on colloidal solutions.

The aim of this study was to evaluate the effect of four methods of sperm selection, on the integrity and stability of the plasma membrane, integrity of the acrosomal membrane and spermatic morphology in frozen/thawed ovine semen. Two types of colloidal silica: colloidal silica-silane and colloidal silica-polyvinylpyrrolidone (PVP), and two aliquots: 1 and 4 ml, were used for sperm selection. Probes FITC-PSA and PI were used to measure the integrity of the plasma and acrosomal membranes. Plasma membrane stability was measured, using fluorescent probes M540 and YOPRO1. Effective reduction in the incidence of spermatozoa with acrosomal pathologies was only achieved using 1 ml colloidal silica-silane. All methods were efficient in select viable and unreacted spermatozoa. Only methods using 1 ml of silica were efficient in decrease spermatozoa stained by PI (death). Methods using silica colloidal-silane were more efficient to decrease apoptotic cells after selection when compared to silica colloidal-PVP. In conclusion, sperm selection in colloidal silica-silane and colloidal silica-PVP improved sperm quality when compared to the controls. The method using 1 ml of colloidal silica-silane is the preferred method because its effectiveness and lower cost.

Viability of ovine spermatozoa collected from epididymides stored at 18°-25°C for 48 hours post mortem / Viabilidade dos espermatozoides ovinos coletados de epidídimos armazenados a 18°-25°C até 48 horas após a morte

The objectives of this study were to verify the time during which viable ovine spermatozoa could be recovered from the cauda epididymis kept at ambient temperature (18-25°C). Sperm collected in an artificial vagina (AV) were used as control. Spermatozoa samples were collected with an AV and from epididymis at 0 (G0), 6 (G6), 12 (G12), 24 (G24), and 48 (G48) hours post mortem. Total motility (TM), progressive motility (PM), hypo-osmotic membrane integrity test (HOST) and morphological changes were assessed. TM decreased (P<0.05) from 24 hours post mortem (70.0±1.9%) compared to AV (86.4±1.0%). PM decreased (P<0.05) from 12 hours after death (31.3±4.0%) compared to AV group (73.2±1.4%). The percentage of viable cells in HOST decreased (P<0.05) in the G48 (60.0±8.9%). Spermatozoa recovery was lower (P<0.05) 48 hours after death (2064.2±230.7 x 106 spermatozoa) compared to G0(2623.6±288.4 x 106 spermatozoa). In conclusion, under the conditions of this study, it would be possible to use epididymal spermatozoa recovered up to 24 hours after death for artificial insemination or in vitro fertilization; however, fertility trials are necessary to prove this hypothesis.(AU)

Os objetivos deste estudo foram avaliar o período pelo qual era possível recuperar espermatozoides ovinos viáveis da cauda de epidídimos mantidos em temperatura ambiente (18-25°C). O sêmen coletado em vagina artificial (AV) foi utilizado como controle. Os espermatozoides foram coletados dos epidídimos à zero hora (G0), às seis (G6), 12 (G12), 24 (G24) e 48 (G48) horas post mortem. A motilidade total (TM), a motilidade progressiva (PM), a integridade de membrana plasmática em solução hiposmótica (HOST) e a morfologia espermática foram avaliadas. A TM diminuiu (P<0,05) a partir de 24 horas após a morte (70,0±1,9%) comparado ao sêmen coletado em AV (86,4±1,0%). A PM diminuiu (P<0,05) a partir de 12 horas após a morte (31,3±4,0%) comparado ao grupo AV (73,2±1,4%). A porcentagem de espermatozoides viáveis no HOST diminuiu (P<0,05) no G48 (60,0±8,9%). A recuperação espermática foi menor (P<0,05) 48 horas após a morte (2064,2±498,1 x 106 espermatozoides) comparado ao G0 (2298,4±288,4 x 106 espermatozoides). Em conclusão, nas condições deste estudo, é possível utilizar espermatozoides epididimários recuperados até 24 horas após a morte para inseminação artificial ou fertilização in vitro, porém testes de fertilidade são necessários para comprovar essa hipótese.(AU)

Relationship of antral follicular blood flow velocity to superovulatory responses in ewes.

The aim of this study was to examine the association between antral follicular blood flow velocity and the response of ewes to hormonal ovarian superstimulation. Ten Santa Inês ewes were subjected to a short- (7days; Group 1) or long-term (13days; Group 2) progesterone (CIDR®; InterAg, Hamilton, New Zealand) priming, and a superovulatory treatment with porcine follicle-stimulating hormone (pFSH; Folltropin®-V; Bioniche Animal Health, Belleville, ON, Canada), given twice daily for four consecutive days in decreasing doses and initiated four or ten days after CIDR insertion, respectively. Embryos were recovered surgically seven days after the last pFSH dose. From one day prior to until the end of the pFSH regimen (Days -1 to 3), all ewes underwent daily transrectal ultrasonography of ovaries. The number of high-velocity pixels (HVPs; 0.055-0.11m/s or upper 50% of recordable velocities) on Day 1 correlated directly with the number of corpora lutea (CL; r=0.92, P=0.0002) and of viable embryos (r=0.77, P=0.01). Correlations were also recorded between the number of HVPs on Day 3 and the recovery rate (r=-0.69, P=0.03), viability rate (r=-0.64, P=0.05), and percentage of degenerated embryos (r=0.65, P=0.04). The percentage of HVPs relative to the total area of ovarian cross section on Day 1 was correlated with the number of CL (r=0.95, P<0.001) and of viable embryos (r=0.85, P=0.002). This parameter on Day 3 was also correlated with the recovery rate (r=-0.69, P=0.03). The percentage of HVPs relative to the total Doppler area on Day 0 was correlated with the recovery rate (r=0.72, P=0.02). It can be concluded that sonographic assessment of high-velocity antral follicular blood flow has the makings of a useful non-invasive method to predict the outcome of the superovulatory treatment in ewes.

Kinematic and spermatic recovery after selection by centrifugation in colloid solutions of ovine cryopreserved semen / Cinética e recuperação espermática após seleção por centrifugação em soluções coloidais de sêmen ovino criopreservado

Frozen and thawed ovine semen undergo morphological and functional changes that prevent or decrease the efficiency of fertilization. Sperm selection methods seek to improve the quality and viability of the fertilizing materials. Four sperm selection methods were employed, using two silica colloidal solutions coated with silane (silica colloidal-silane) or by polyvinylpyrrolidone (silica colloidal-PVP), and varying the volume of colloidal solution. Sperm kinematic and sperm recovery were evaluated by means of CASA. The protocols using silica colloidal-silane showed higher total motility (TM), progressive motility (PM) and percentage of rapid sperm (%RAP) compared to the methods employing silica colloidal-PVP and to the samples prior to sperm selection. The silica colloidal-PVP had greater sperm recovery compared to the silica colloidal-silane. Only the method using 4mL of silica colloidal-PVP was not efficient in selecting samples with better quality compared to the samples analyzed prior to sperm selection. The methods using lower volumes of colloidal solution did not differ from those using higher volumes and the best results were shown by the method with 1mL silica colloidal-silane. The results found in the study indicated greater efficiency of the silica colloidal-silane solution for sperm selection of thawed ovine semen when compared to selection using silica colloidal-PVP. The method using 1mL of silica colloidal-silane was equally efficient to the method with higher volume, presenting itself as an alternative to process samples with lower sperm concentration.(AU)

O sêmen ovino congelado e descongelado sofre alterações morfofuncionais que impossibilitam ou diminuem a eficiência na fecundação. Os métodos de seleção espermática visam melhorar a qualidade e a viabilidade do material fecundante. Foram utilizados quatro métodos de seleção espermática utilizando duas soluções de sílica coloidal revestida por silano (sílica coloidal-silano) ou por polivinilpirrolidona (sílica coloidal-PVP), variando o volume de solução coloidal. Foram testadas a cinética espermática no CASA e a recuperação espermática. Os protocolos utilizando sílica coloidal-silano apresentaram maior motilidade total (MT), motilidade progressiva (MP) e porcentagem de espermatozoides rápidos (% RAP) quando comparados aos métodos utilizando a sílica coloidal-PVP e às amostras antes da seleção espermática. A sílica coloidal-PVP teve maior recuperação espermática quando comparada à sílica coloidal-silano. Somente o método utilizando 4mL de sílica coloidal-PVP não foi eficiente na seleção de amostras com melhor qualidade quando comparado às amostras analisadas antes da seleção espermática. Os métodos utilizando menores volumes de solução coloidal não diferiram dos métodos de maior volume, sendo a sílica coloidal-silano com 1mL o método que apresentou os melhores resultados. Como conclusão, os resultados encontrados no trabalho apontaram a maior eficiência da sílica coloidal-silano em selecionar sêmen ovino congelado e descongelado quando comparado à seleção em sílica coloidal-PVP. O método utilizando 1mL de sílica coloidal-silano foi igualmente eficiente ao método com maior volume, sendo uma alternativa para processar amostras com baixa concentração espermática.(AU)

Detecting animal by-product intake using stable isotope ratio mass spectrometry (IRMS).

Sheep are used in many countries as food and for manufacturing bioproducts. However, when these animals consume animal by-products (ABP), which is widely prohibited, there is a risk of transmitting scrapie - a fatal prion disease in human beings. Therefore, it is essential to develop sensitive methods to detect previous ABP intake to select safe animals for producing biopharmaceuticals. We used stable isotope ratio mass spectrometry (IRMS) for 13C and 15N to trace animal proteins in the serum of three groups of sheep: 1 - received only vegetable protein (VP) for 89 days; 2 - received animal and vegetable protein (AVP); and 3 - received animal and vegetable protein with animal protein subsequently removed (AVPR). Groups 2 and 3 received diets with 30% bovine meat and bone meal (MBM) added to a vegetable diet (from days 16-89 in the AVP group and until day 49 in the AVPR group, when MBM was removed). The AVPR group showed 15N equilibrium 5 days after MBM removal (54th day). Conversely, 15N equilibrium in the AVP group occurred 22 days later (76th day). The half-life differed between these groups by 3.55 days. In the AVPR group, 15N elimination required 53 days, which was similar to this isotope's incorporation time. Turnover was determined based on natural 15N signatures. IRMS followed by turnover calculations was used to evaluate the time period for the incorporation and elimination of animal protein in sheep serum. The δ13C and δ15N values were used to track animal protein in the diet. This method is biologically and economically relevant for the veterinary field because it can track protein over time or make a point assessment of animal feed with high sensitivity and resolution, providing a low-cost analysis coupled with fast detection. Isotopic profiles could be measured throughout the experimental period, demonstrating the potential to use the method for traceability and certification assessments.

Follicular wave emergence in Santa Inês ewes subjected to long-term, progesterone-based estrous synchronization protocols at different times of the year.

This study was conducted to document the pattern of antral follicular wave emergence throughout the 14-day, progesterone (P4)-based estrous synchronization protocol in ewes that were maintained in subtropical conditions, during the period of increasing day lengths (ID), decreasing day lengths (DD), and the transitional period (TP). In addition, the influence of ovarian status (i.e., size of ovarian antral follicles and the presence of corpora lutea) at the outset of P4 treatment on ensuing ovarian follicular wave development was examined. Sexually mature Santa Inês ewes (n=70) were subjected to one of the two estrous synchronization protocols in the three seasons. On Day 0, the ewes received an i.m. injection of prostaglandin F2α and an intravaginal P4-releasing device that remained in place for 14days (G-1CIDR) or was replaced on Day 7 (G-2CIDR). Daily ultrasonography of ovaries was conducted from Days 0 to 15. Mean (±SEM) numbers of follicular waves per ewe were 3.7±0.1 and 3.6±0.1 for G-1CIDR and G-2CIDR (P>0.05). The number of emerging follicular waves was greater (P<0.05) during the ID period than during the TP and DD periods (4.0±0.1, 3.4±0.1 and 3.6±0.1, respectively). The presence of medium-sized antral follicles (4.0 to 5.75mm) in the absence of corpora lutea at the time of CIDR insertion tended to advance follicular wave emergence. Although the long-term P4 treatment was not originally designed to synchronize follicular waves, there was a distinctive pattern of follicular wave dynamics during the period of application of CIDRs that was affected mainly by the number of emerging follicular waves and ovarian status at CIDR insertion.

In vitro Developmental Competence of Adult Sheep Oocytes Treated with Roscovitine.

The efficiency of in vitro sheep embryo production is still low compared to that observed in vivo and in other species. In this context, meiotic inhibition strategies emerged as a promising alternative to improve this biotechnology. So, this study aimed to evaluate, for the first time, the effects of roscovitine on in vitro maturation of sheep oocytes and their subsequent embryo development. For this, cumulus-oocyte complexes (COCs) were cultured for 6 h in the presence (Rosco) or absence (Control) of 75 µm roscovitine and, subsequently, in vitro matured (IVM) for 18 h with gonadotropins. At 0 (Immature), 6 and 24 h of culture, the nuclear status of oocytes was evaluated by Hoechst staining. Embryo cleavage and blastocyst formation were recorded 30 h after in vitro fertilization and on day 7 of culture, respectively. Blastocyst quality was evaluated by differential staining. At 6 h, the GV rate in the Rosco treatment (93.8%) was similar to that observed in the Immature oocytes (94.9%) and significantly higher compared to Control (41.3%). After IVM for 18 h, a high and similar proportion of oocytes from Rosco (93.6%) and Control (88.4%) reached the MII stage. In both treatments, approximately 70% of oocytes cleaved and 50% of them developed up to blastocyst. The mean percentage of blastocyst cells, embryoblast, trophoblast and pyknosis did also not differ between Control and Rosco. In conclusion, roscovitine, at the studied experimental conditions, was efficient to reversibly inhibit the meiosis of adult sheep oocytes without detrimental effect on development and quality of the in vitro produced embryos.

Effects of season and ovarian status on the outcome of long-term progesterone-based estrus synchronization protocols and ovulatory follicle development in Santa Inês ewes under subtropical conditions.

This study set out to investigate the efficiency of long-term estrus synchronization protocols and ovulatory follicle dynamics in ultrasonographically monitored Santa Inês ewes during lengthening (LD; September-October) and shortening photoperiods (SD; April-May), and the transitional period (TP; January). In addition, the influence of ovarian status (e.g., size of antral follicles and/or presence of corpora lutea) at the outset of the estrus synchronization protocols on the ensuing development of ovulatory follicles was examined. Seventy sexually mature Santa Inês ewes were subjected to one of the two estrus synchronization regimens; on Day 0 (random day of the estrous cycle or anovulatory period), the ewes were fitted with an intravaginal progesterone (P4)-releasing (controlled intrauterine drug release [CIDR]) device, which was left in place for 14 days (G-1CIDR, n = 35) or replaced on Day 7 (G-2CIDR, n = 35), and received an intramuscular injection of 10 mg of PGF2α. The ewes allocated to the G-1CIDR group had mean serum P4 concentrations less than 2 ng/mL during the last 4 days of the synchronization protocol. There were no differences (P > 0.05) in mean ovulation rates between the two protocols tested nor among the ewes varying in ovarian status or studied at different times of the year, but ovulations occurred ∼ 12 hours later in the TP compared with the SD period (P < 0.05). Ovulatory follicles emerged earlier (P < 0.05) in the G-1CIDR group than in the G-2CIDR group (Day 8.3 ± 0.5 vs. 9.2 ± 0.4) and during LD (Day 7.1 ± 0.6) compared with the TP (Day 9.1 ± 0.6) and SD (Day 9.9 ± 0.5 of the protocol). In conclusion, the replacement of CIDR devices prevented the occurrence of lower-than-normal luteal phase levels of P4 at the end of the 14-day estrus synchronization protocol. However, although this procedure and seasonal influences altered certain growth characteristics of ovulatory follicles, there were no effects of these factors on the mean ovulation rate.

Effect of oil overlay on inhibition potential of roscovitine in sheep cumulus-oocyte complexes.

Inhibitors of cyclin-dependent kinases, as roscovitine, have been used to prevent the spontaneous resumption of meiosis in vitro and to improve the oocyte developmental competence. In this study, the interference of oil overlay on the reversible arrest capacity of roscovitine in sheep oocytes as well as its effects on cumulus expansion was evaluated. For this, cumulus-oocyte complexes (COCs) were cultured for 20 h in TCM 199 with 10% foetal bovine serum (Control) containing 75 µm roscovitine (Rosco). Subsequently, they were in vitro matured (IVM) for further 18 h in inhibitor-free medium with LH and FSH. The culture was performed in Petri dishes under mineral oil (+) or in 96 well plates without oil overlay (-) at 38.5°C and 5% CO2 . At 20 and 38 h, the cumulus expansion and nuclear maturation were evaluated under stereomicroscope and by Hoechst 33342 staining, respectively. No group presented cumulus expansion at 20 h. After additional culture with gonadotrophins, a significant rate of COCs from both Control groups (+/-) exhibited total expansion while in both Rosco groups (+/-) the partial expansion prevailed. Among the oocytes treated with roscovitine, 65.2% were kept at GV in the absence of oil overlay while 40.6% of them reached MII under oil cover (p < 0.05). This meiotic arrest was reversible, and proper meiosis progression also occurred in the Control groups (+/-). So, the culture system without oil overlay improved the meiotic inhibition promoted by roscovitine without affecting the cumulus expansion rate or the subsequent meiosis progression.

Identification of the SP22 sperm protein in Santa Inês and Dorper rams.

The sperm membrane protein referred to as SP22 has been identified in different species and, at least in rats, is highly correlated with fertility. The goals of this study were to identify and to quantify the SP22 protein on spermatozoa from adult rams (Dorper and Santa Inês breeds), and to correlate its levels to morphological and kinematics parameters. SP22 on ram sperm was effectively quantified by both enzyme-linked immunosorbent assay (ELISA) and fluorescein isothiocyanate immunostaining analysis and the two methods were significantly correlated (R(2) = 0.70). Clustering analysis of motility parameters obtained by computer-assisted semen analysis system was used to establish that three distinct kinematic subpopulations with different vigour and progressiveness coexistent within ejaculate. While there were significant differences in the distribution of the three subpopulations in the rams, there was no significant correlation between the proportion of each subpopulation in the rams and the SP22 levels. Quantification of SP22 immunostaining intensity was not correlated with any of the sperm parameters. However, SP22 levels obtained by ELISA were negatively correlated with morphological abnormalities and positively correlated with membrane integrity (three variable R(2) = 0.47). Future breeding studies are now needed to validate that this protein is a biomarker of fertility in this species.

Tritrichomonas fetus extracellular products decrease progressive motility of bull sperm.

Tritrichomonas fetus causes infertility and abortion in cattle; however, there is scarce information regarding the susceptibility of bovine sperm to this parasite. The objective of this study was to analyze in vitro the interaction between T. fetus and bovine sperm and to evaluate the effect of extracellular products secreted by the parasite on these reproductive cells. Sperm from five fertile bulls (Bos taurus taurus, Holstein-Friesian), selected through a Percoll gradient, adhered to T. fetus after 30min of interaction, resulting in agglutination between the two kinds of cells. Based on reverse transcription-polymerase chain reaction (RT-PCR), T. fetus continuously expressed its gene for cysteine peptidase in the presence or absence of sperm. Computer-assisted semen analysis (CASA) revealed that, after 1h incubation of sperm in T. fetus culture extract, the extracellular products secreted by the parasite decreased sperm progressive motility (P<0.05). Although T. fetus extracellular products did not lead to loss of sperm viability (P<0.05) based on the Annexin-V/propidium iodide assay, the percentage of Annexin-V fluorescein isothiocyanate-positive and propidium iodide-positive cells increased (P<0.05) during incubation of sperm in T. fetus culture extract, consistent with cellular damage. In conclusion, extracellular products secreted by T. fetus were cytotoxic to bovine sperm, as they decreased sperm progressive motility; perhaps this contributes to the pathogenesis of T. fetus-induced infertility.

Viability of OPS vitrified sheep embryos after direct transfer.

The aim of this study was to evaluate the viability in the effect of open pulled straw (OPS) vitrification procedure of sheep embryos after direct transference. Embryos were produced in vivo and cryopreserved in slow freezing or OPS vitrification. The survival rates of cryopreserved embryos were compared to non-frozen standard pattern. In a first set of experiments, embryos at morula and blastocyst stages were dived in ethylene glycol (1.5 M) and frozen in an automatic freezer. After being thawed, they were directly or indirectly transferred to ewes recipient. A second group of embryos were drawn into OPS and plunged into liquid nitrogen after being exposed at room temperature for 1 min and 45 s in 10% EG plus 10% dimethyl sulphoxide (DMSO), then again for 30 s in 20% EG + 20% DMSO + 0.5 M sucrose. After being warmed, embryos were also directly transferred using a French mini straw as the catheter for the transplantation process or after in vitro dilution of cryoprotectants (two-step-process). No significant difference was observed among fresh, frozen or vitrified embryos on pregnancy rate (50.0%, 38.6% and 55.8%). However, when we evaluated only the direct transference, the pregnancy rate of OPS vitrified embryos was higher than that of frozen embryos (57.1% vs 34.8%) (p = 0.07). In addition, vitrified morulae had a higher pregnancy rate than the one with frozen embryos (64.0% vs 38.9%) (p = 0.07). Finally, our results indicate that OPS vitrification technique in association with direct transference improves the viability of sheep embryos with potential applications to field conditions.

Concentrações plasmáticas de testosterona, triiodotironina (T3) e tiroxina (T4) em bodes submetidos ao estresse calórico / Plasma concentrations of testosterone, triiodothyronine (T3), and thyroxine (T4) in bucks submitted to heat stress

Para verificar o efeito do estresse calórico (EC) nas concentrações plasmáticas de testosterona, triiodotironina (T3) e tiroxina (T4), oito bodes, das raças Saanen (n=4) e Alpina (n=4), foram mantidos em câmara bioclimática, sob condições de termoneutralidade (13,0ºC a 26,7ºC) durante 30 dias e, após um período (60 dias) de descanso, submetidos ao EC (23,7ºC a 34,0ºC) por 30 dias. Para minimizar as variações sazonais nos perfis hormonais devido ao fotoperíodo, durante toda fase experimental, incluindo a de adaptação em condições de termoneutralidade (30 dias), o fotoperíodo foi controlado utilizando-se alternância de dias longos (16h de luz e 8h de escuro) e de dias curtos (8h de luz e 16h de escuro) a cada 30 dias. As amostras de sangue foram coletadas duas vezes por semana durante cinco semanas. No conjunto das raças, o EC não influenciou (P>0,05) as concentrações de testosterona (1,8±0,2 vs 1,3±0,2ng/ml) e nem a de T4 (52,7±2,8 vs 50,0±2,8ng/ml). Houve declínio (P<0,01) das concentrações de T3 nos animais submetidos ao experimento (1,3±0,1 vs 1,0±0,1ng/ml), mas a redução foi observada somente nos bodes Saanen. Em ambas as raças, as concentrações de T3 e T4 variaram (P<0,01) conforme o dia da coleta das amostras de sangue. O EC foi suficiente para produzir uma resposta fisiológica com redução das concentrações plasmáticas de T3 em bodes das raças Saanen, mas não da raça Alpina, assim como não foi capaz de alterar os níveis plasmáticos de testosterona e nem de T4.

To verify the effect of heat stress (HS) on plasma testosterone, triiodothyronine (T3), and thyroxine (T4) concentrations, eight Saanen (n=4) and Alpine Brown (n=4) bucks were kept in climate chamber under thermal neutral conditions (13.0ºC to 26.7ºC) for 30 days. After a resting period (60 days), the same bucks were submitted to heat stress (23.7ºC to 34.0ºC) for another 30 days. To neutralize the seasonal variations of hormonal profiles throughout the period, the photoperiod was controlled every 30 days altering long (16 hours of light and 8 hours of darkness) and short days (8 hours of light and 16 hours of darkness). The blood samples were collected twice a week during five weeks. In both breeds, there was no effect of HS (P>0.05) on plasma concentrations of testosterone (1.8±0.2 vs 1.3±0.2ng/ml) and T4 (52.7±2.8 vs 50.0±2.8ng/ml). There was a decline (P<0.01) of plasma T3 concentrations (1.3±0.1 vs 1.0±0.1ng/ml) after HS treatment, but this reduction was only evident in Saanen bucks. In both breeds, the plasma concentrations of T3 and T4 varied (P<0.01) according to the day of blood sample collection. The HS was sufficient to provoke a physiological response with reduction of plasma concentrations of thyroid hormones mainly of T3 in Saanen bucks, but not in Alpine ones. The HS did not affect the plasma testosterone and T4 levels.

An unusual gangrenous goat mastitis caused by Staphylococcus aureus, Clostridium perfringens and Escherichia coli co-infection

Relata-se, pela primeira vez no Brasil, a ocorrência de mastite gangrenosa caprina atípica causada pela co-infecção por Staphylococcus aureus, Clostridium perfringens e Escherichia coli em uma cabra da raça Boer, na segunda semana de lactação. Descrevem-se os achados clínicos, os procedimentos de diagnóstico microbiológico e a conduta terapêutica.

Determinação dos ritmos circadianos e circanuais nas concentrações plasmáticas de testosterona e androstenediona em machos ovinos / Circadian and circannual rhythms determination in testosterone and androstenedione plasma concentrations in ovine males / Determinación de los ritmos circadianos y circanuales en las consentraciones plasmáticas de testotesrona y androstenediona en machos ovinos

Estudou-se a estacionalidade reprodutiva de machos ovinos no Estado de São Paulo para estabelecer os ritmos endócrinos circadianos e circanuais, de cinco carneiros Ideal-Polwarth. O sangue foi colhido por venopunção jugular, em um período de 24 horas, sendo as colheitas realizadas de duas em duas horas, a cada dois meses (seis momentos de colheita em um ano), para a obtenção do plasma e avaliação dos níveis de testosterona e androstenediona por radioimunoensaio. As concentrações plasmáticas de androstenediona e testosterona não permitiram a definição de um ritmo circadiano em suas respectivas secreções, porém, demonstraram ritmos circanuais, com níveis mais baixos de junho a outubro e mais elevados de dezembro a abril.

The reproductive seasonality of ovine males in Sao Paulo State was studied to establish the circadian and circannual endocrine rhythms of 5 Ideal-Polwarth rams. Blood was collected by puncturing the jugular vein for 24 hours with 2-hour intervals every 2 months (6 collections throughout the year) in order to obtain plasma and evaluate testosterone and the androstenedione levels by radioimmunoassay. The Testosterone and androstenedione plasma concentrations did not demonstrate a circadian rhythm in their respective secretions; however, they showed lower levels of circannual rhythms from June to October and higher from December to April.

Se estudió la estacionalidad reproductiva en machos ovinos en el Estado de São Paulo para determinar los ritmos endocrinos circadianos y circanuales, de cinco moruecos Ideal-Polwarth. La sangre se recogió por venopunción yugular, en un período de 24 horas, con las colectas de dos en dos horas, a cada dos meses (seis colectas en un año), para la obtención del plasma y evaluación de los niveles de la testosterona y androstenediona por el radioimunoensayo. Las concentraciones de la testosterona y androstenediona no permitieron la definición de un ritmo circadiano en sus secreciones, pero demostraron ritmos circanuales, con niveles más bajos de junio hasta octubre y más elevados de diciembre hasta abril.

Diagnóstico ultra-sonográfico de piometra em cadelas / Ultrasonographic diagnosis of pyometera in bitches

A ultra-sonografia do tipo B (SCANNER 450 (5MHz), Pie Medical, Holanda) foi utilizada isolada ou em conjunto com exames laboratoriais e radiográficos em 33 cadelas com história clínica compatível com piometra. O útero dilatado apresentou imagem ultra-sonográfica de uma estrutura tubular bem definida com diâmetro entre 0,5 e 4,0 cm. O conteúdo luminal uterino apresentou menor ecogenicidade que a parede, com cintilaçöes ecogênicas bem evidentes. Houve concordância entre o aumento da viscosidade da secreçäo e a intensidade ecogênica. O diagnóstico ultra-sonográfico foi possível em 31 animais (94 por cento), e foi confirmado através de laparotomia ou necrópsia. Concluímos que a ultra-sonografia do tipo B é método eficiente, podendo ser utilizado no diagnóstico da piometra canina

Estudo da técnica de coleta, congelaçäo e descongelaçäo de embriöes de caprinos (Capra hircus), da raça Saanen, portadores da translocaçäo 5/15 / Study of embryos colect technique, frozen and thawing of Saanen (Capra hircus), goats bearing 5/15 translocation

No presente trabalho foi realizado um estudo da técnica cirúrgica de coleta de embriöes na espécie caprina, utilizando-se doadoras heterozigotas para a translocaçäo 5/15, acasaladas com animal translocado. Simultaneamente foram observados aspectos técnicos inerentes à criopreservaçäo, descongelaçäo e cultivo das estruturas recuperadas. Frente ao tratamento superovulatório, o cio foi observado em 24 ou 48 horas após a remoçäo da esponja intravaginal e foram recuperadas 71 estruturas viáveis em 43 coletas. O 1-2 Propanodiol mostrou-se eficiente criopreservador, permitindo cultivo de todos os embriöes, fato näo observado quando utilizou-se o Glicerol. As aderências provocadas pelas repetidas coletas constituem-se no grande desafio no sentido de se aperfeiçoar a técnica, pois säo consideradas um fator limitante ao uso de doadores no início da vida reprodutiva

Modificaçöes na técnica de correçäo cirúrgica de dilaceraçäo perineal de 3§ grau em éguas / Modifications of surgical correction of the 3rd degree perineal laceration in mares

Em 52 éguas de diferentes raças e idades, praticou-se cirurgia corretiva de dilaceraçäo perineal de 3§ grau, com modificaçöes na técnica de GOTZE, na tentativa de restabelecer a anatomia do sistema reprodutor, bem como a vida reprodutiva destas fêmeas. Das éguas que sofreram a cirurgia, a totalidade (100//) retornou à reproduçäo, alcançando-se o índice máximo de fertilidade após inseminaçäo artificial ou cobertura. Os resultados obtidos pelo emprego da técnica proposta permite indicá-la como uma conduta correta e eficiente na recuperaçäo anátomo-funcional do genital de éguas com dilaceraçäo de períneo de 3§ grau

A practical approach for the treatment of bovine trichomoniasis

Cinco touros, naturalmente infectados com Trichomonas foetus, cada um pesando 800 kg e com idade entre seis e oito anos, foram tratados, por via intravenosa, com uma única aplicaçäo de benzoilnitroimidazole a 50 mg/kg e diluídos em três litros de soluçäo de 20 por cento de DMSO. A pesquisa microscópica a fresco e o cultivo microbiológico do protozoário revelaram-se negativos às 24 horas e 12 semanas pós-tratamento. Os touros tratados emprenharam fêmeas que levaram a gestaçäo a termo. O êxito obtido nesse procedimento terapêutico pode representar, a campo, praticidade no controle de trichomoniase bovina