Romildo Romualdo Weiss

Association of antibodies against Neospora caninum in mares with reproductive problems and presence of seropositive dogs as a risk factor.

Sera from 112 mares from 5 horse-breeding farms was examined for the presence of antibodies to Neospora caninum and Toxoplasma gondii by an indirect fluorescent antibody test (IFAT), as well as from dogs and cattle present on these properties for the presence of antibodies to N. caninum. Among the 112 mares, 35 had a history of reproductive problems in the last breeding season and 77 had no reproductive problems. The rates of seroprevalence of N. caninum in mares with and without a history of reproductive problems were 25.71% and 6.49% and from T. gondii 2.85% and 1.29%, respectively. In dogs and cattle, the rates of seroprevalence of N. caninum were 10.52% and 15.55%, respectively. A positive correlation was found between the presence of antibodies against N. caninum (p=0.010) in mares and the occurrence of reproductive problems using the Fishers exact test. Significantly higher seroprevalence for N. caninum in mares was observed on the farm that had seropositive dogs (p=0.018). Cattle on this farm were also seropositive. No significant differences in seropositivity were found on farms where dogs were seronegative, or absent. This result suggests, for the first time, the presence of seropositive dogs as a risk factor for N. caninum in mares and the necessity for further investigation of the epidemiology of this parasite in horse-breeding farms with reproductive problems and the presence of cattle and dogs. This is the first report on the occurrence of antibodies against N. caninum in horses from the state of Santa Catarina, Brazil.

Comparison between deep intracornual artificial insemination (dIAI) and conventional artificial insemination (AI) using low concentration of spermatozoa in beef cattle

This study aimed to compare the pregnancy rate using the conventional artificial insemination (AI) or deep intracornual artificial insemination (DIAI), with low number of spermatozoa (4.0 million sperm) in 270 Nelore cows. The animals were divided in two groups (G: G1 (135 cows) conventional AI was performed (=semen deposition in the uterine body) and in G2 (135 cows) to DIAI, in ipsilateral horn where the dominant follicle in the ovary had previously been detected, by ultrasound examinations. For both the methods, a single artificial insemination was carried out after visual estrus observation, checked three times a day (morning, afternoon and evening). The pregnancy diagnosis after 45 days was conducted by ultrasound. Results showed a better pregnancy rate in the DIAI group (67.4% - p<0.01), when compared to conventional AI (48.8%) with low spermatozoa concentration.

Viability of bull spermatozoa collected from the epididymis stored at 18-20°C

The aim of this study was to evaluate the viability of bull spermatozoa collected from the cauda epididymis stored at 18-20°C, which were compared with semen collected by electro-ejaculation method and preserved at 5°C. Ten pairs of testes from Tabapua bulls were removed by orchiectomy and stored for 6 (G6), 12 (G12), 18 (G18), 24 (G24) and 30 (G30) h at room temperature (18-20°C). Seven days before orchiectomy, semen was collected by electro-ejaculation method. The sperm parameters evaluated were: sperm motility, vigor, concentration, morphology and acrosome defects. Sperm motility declined (p<0.05) when spermatozoa were stored for 30 h in the epididymis. The spermatozoa from the epididymis showed lower sperm motility than that of spermatozoa collected via electro-ejaculation. There was a little expressive decrease in sperm vigor and increased in morphological defects with storage time, but the acrosome integrity was not affected. Cold storage (5°C) maintained sperm viable for 15 to 40.8 h. Thus, it was possible to recover viable sperm with 41.25% of motility from the cauda epididymis stored at room temperature of 18-20°C for 30 h. There were differences between the ejaculated and epididymal sperm for the bulls and the conservation at 5°C allowed short-term preservation of the gametes.

Kinematic and spermatic recovery after selection by centrifugation in colloid solutions of ovine cryopreserved semen

O semen ovino congelado e descongelado sofre alteracoes morfofuncionais que impossibilitam ou diminuem a eficiencia na fecundacao. Os metodos de selecao espermatica visam melhorar a qualidade e a viabilidade do material fecundante. Foram utilizados quatro metodos de selecao espermatica utilizando duas solucoes de silica coloidal revestida por silano (silica coloidal-silano) ou por polivinilpirrolidona (silica coloidal-PVP), variando o volume de solucao coloidal. Foram testadas a cinetica espermatica no CASA e a recuperacao espermatica. Os protocolos utilizando silica coloidal-silano apresentaram maior motilidade total (MT), motilidade progressiva (MP) e porcentagem de espermatozoides rapidos (% RAP) quando comparados aos metodos utilizando a silica coloidal-PVP e as amostras antes da selecao espermatica. A silica coloidal-PVP teve maior recuperacao espermatica quando comparada a silica coloidal-silano. Somente o metodo utilizando 4mL de silica coloidal-PVP nao foi eficiente na selecao de amostras com melhor qualidade quando comparado as amostras analisadas antes da selecao espermatica. Os metodos utilizando menores volumes de solucao coloidal nao diferiram dos metodos de maior volume, sendo a silica coloidal-silano com 1mL o metodo que apresentou os melhores resultados. Como conclusao, os resultados encontrados no trabalho apontaram a maior eficiencia da silica coloidal-silano em selecionar semen ovino congelado e descongelado quando comparado a selecao em silica coloidal-PVP. O metodo utilizando 1mL de silica coloidal-silano foi igualmente eficiente ao metodo com maior volume, sendo uma alternativa para processar amostras com baixa concentracao espermatica.

The effects of intramuscular or intravenous injections of gonadotropin releasing hormone at fixed-time artificial insemination on pregnancy rates of Bos indicus beef cows

The effect of an intramuscular versus intravenous administration of gonadotropin releasing hormone (GnRH) at fixed-time AI (FTAI) on the pregnancy rates of crossbred Bos indicus beef cows was evaluated. Pluriparous nursing calv cows (n=120) were synchronized as follows: d 0 cows received a 2.0 mg injection of estradiol benzoate (EB) and insertion of a controlled intravaginal progesterone releasing device containing 0.558 g of progesterone, d 8 removal of the progesterone device , a 0.15 mg injection of prostaglandin F2α (PGF), a 1.0 mg injection of EB, and 400 IU injection of equine chorionic gonadotropin. Fifty-four hr after PGF, all cows were exposed to FTAI and a 0.084 mg injection of GnRH was administered either via Vena caudalis (n=60), or via Longissimus dorsi (n=60). Cows were inseminated with the same sire and by a single AI technician. Pregnancy was determined by the transrectal ultrasonography on d 40 after AI. Cows receiving the intravenous administration of GnRH had higher (P = 0.04) pregnancy rates than the cows receiving the intramuscular injection of GnRH (65 vs 46.6%, respectively). It was concluded that the intravenous administration of GnRH at the time of AI improved the pregnancy rates of crossbred Bos indicus beef cows submitted to FTAI.

Embryo transfer in no cycling Crioula and Quarter horse breeds treated with estradiol cipionate and long-acting progesterone

The aim of this work was to prepare the mares for embryo transfer. In group 1 (G1,treated, n=15), recipient mares in anoestrus or in a transition period were treated with 5.0, 3.0 and 2.0 mg of estradiol cipionate at the days 0, 1 and 2 respectively, beginning at the day of ovulation (DO). From the fourth day on, the mares this group received long-acting progesterone weekly, up to the 120 day of gestation. At D8, the embryo was collected from the donor and transferred to the recipient. At D12, the ultrasonographyc diagnosis of pregnancy was carried out. The control group (G2, not treated, n=20) was formed by cycling recipient mares, displaying ovulation at each 2 to 3 days after the donors mare ovulation. The pregnancy rate was higher (p<0.05) in the mares from G2 (85.0%) than from G1 (53.3%). Thus, it could be concluded that the treated mares although showed lesser pregnancy rate than the cycling mare, were satisfactory alternative to be used mainly when there is no available cycling recipient.

Estrus synchronization and Fixed Time Artificial Insemination (FTAI) in dairy buffaloes during seasonal anestrus

The aim of this work was to study estrus synchronization and fixed time artificial insemination (FTAI) in dairy buffaloes during season anestrus. One hundred thirty-nine dairy buffaloes in seasonal anestrus were divided in two groups as G1(n=66) and G2(n=73). The protocols for both the groups were the same until day (D)14:D0 administration of 2.0 mg estradiol benzoate and implantation of progesterone device (P4) for 14 days; D14 removal of P4 plus 150 mg of cloprostenol and 400 IU of equine chorionic gonadotropin. On D16, G1 received 10 mg of buserelin and G2 100 mg deslorelin acetate. On D17, both the groups were submitted to FTAI. Ultrasonographic examinations of ovaries were performed on D0, D14, D16 and D17. Results showed that pregnancy rates in G1 and G2 were 20 and 41% (p 0.05). Thirty-five percent of the animals in G1 and 54.1% in G2 showed a diameter DF greater than 8.0 mm on D16 (p>0.05). Thus, it could be concluded that the protocols synchronized the estrus, leading the concentration of the parturitions in the period of low milk production. Deslorelin was more efficient than buserelin due the higher percentage of DF ovulation and higher pregnancy rates.

Determination of progesterone concentration during the estrous cycle in dairy cows using a chemiluminescence assay

The study aimed to determine the patterns of serum progesterone concentration in estrous cycle in dairy cows by a chemiluminescence assay (CLIA). Four non-lactating multiparous Jersey cows were used. Animals with a corpus luteum (CL) in any of the ovaries were induced into estrus. Day zero (d0) of the estrous cycle was defined as the day of visible estrus. Blood samples were collected and ultrasonography (US) of the ovaries were performed until a new manifestation of visible estrus was observed. The lengths of the estrous cycles (estrus to estrus) of the four cows were 20, 21, 22, and 23 days. The mean serum concentrations of P4 (x ± s) were 2.8 ± 1.4 ng/mL in proestrus, 2.4 ± 1.5 ng/mL in estrus, 2.0 ± 1.8 ng/mL in metestrus, and 11.9 ± 5.7 ng/mL in diestrus. The follicular and luteal phases of the estrous cycle were established based on P4 concentrations. P4 serum concentrations ≥5.48 ng/mL indicated the presence of functional CL, which was observed from d3 to d12 of the cycle. P4 concentrations decreased from d13 until next estrus. Thus, the previously mentioned P4 serum concentration was established as the limit for a predominantly functional CL. P4 concentrations < 5.48 ng/mL indicated a nonfunctional CL or CL that is not yet fully formed, which is observed in metestrus. A P4 standard curve, constructed based on the Lorentz distribution, was used to determine values of < 5.48 ng/mL of serum P4 concentration for the follicular phase and ≥ 5.48 ng/mL for the luteal phase. Data obtained from US examinations were consistent with P4 concentrations determined using CLIA. In conclusion, the automated CLIA was efficient in determining the P4 concentrations during the various stages of estrous cycles in dairy cows. The findings of the study will help researchers in the animal reproduction field. We recommend the use of CLIA because it´s available in many laboratories worldwide with the ability to process thousands of samples per day.

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.

Reproductive Performance of Prebubertal and Pubertal Heifers Submitted to Timed Artificial Insemination Protocols

High numbers of prepubertal heifers at the beginning of the breeding season (BS) reduces reproductive efficiency in beef cattle.1 In Brazil, grazing Bos indicus heifers reach puberty between 22 and 36months, and the age at first calving is between 44 and 48months.2 Seasonality, poor pasture management, and poor nutrition during the heifer’s growth phase are some of the variables that lead to the delayed onset of puberty in Bos indicus cattle.3 Studies have been conducted to induce puberty in heifers, using either nutritional management strategies,4 or intravaginal progesterone (P4) devices.5 Sales et al.6 induced prepubertal Bos indicus heifers by administering P4 with estradiol cypionate (ECP) after 10days. Silveira et al.5 used prepubertal and pubertal zebu heifers for timed artificial insemination (TAI). They found a higher pregnancy rate (PR) in the groups that had received P4 with estradiol benzoate (EB) and ECP compared to prepubertal heifers only exposed to bulls. The exposure of prepubertal heifers to P4 for 8days induced and synchronized estrus. Similar effects were observed by Wheaton et al.7 and Demeterco et al.8 Treatment with a P4 intravaginal device induced prepubertal heifers to cycle and also increased the uterine diameter6 even during anestrus.9 The addition of EB at the time of P4 removal potentiates this action.3