Dimethylformamide Preserves the Integrity of Cryopreserved Goat Semen in a Soybean Lecithin-Based Extender.

This study investigated the cryoprotectant effects of dimethylformamide (DMF), ethylene glycol (EG), and dimethyl sulfoxide (DMSO) as substitutes for glycerol (GLY) in a soybean lecithin (SL)-based extender in the cryopreservation of buck sperm. In this study, the semen of three Saanen bucks was individually extended in SL supplemented with 5% GLY (control), DMF, EG, or DMSO. After this, the extended semen was cryopreserved and two straws from each group were thawed (37°C for 30 seconds), pooled, and analyzed for sperm motion parameters, plasma membrane integrity (PMI), acrosomal integrity (ACI), and high mitochondrial membrane potential (HMMP). Samples were analyzed after 15 minutes (T0) and after 2 hours of incubation at 37°C (T2). The results revealed higher values of motility (total and progressive) and sperm motion parameters for DMF than the other cryoprotectants (p < 0.0001). PMI and HMMP did not differ (p > 0.05) between GLY and DMF, but ACI was higher (p < 0.01) for DMF compared with GLY. Based on these results, DMF and GLY samples were used in heterologous in vitro fertilization assays by using bovine oocytes (n = 337) obtained from a slaughterhouse. No differences (p > 0.05) were observed between GLY and DMF for unfertilized (GLY: 38.8%; DMF: 25.33%), pronucleus (GLY: 25.68%; DMF: 27.92%), and cleavage rates (GLY: 35.52%; DMF: 46.75%). Based on these results, it is concluded that DMF preserves sperm motion characteristics and ACI better than GLY, EG, and DMSO, and it is the penetrating cryoprotectant of choice for the cryopreservation of buck sperm in SL extender.

Effects of adiponectin during in vitro maturation of goat oocytes: MEK 1/2 pathway and gene expression pattern.

Recent studies have shown that adiponectin, an adipokine predominantly produced by adipose tissue, regulates several reproductive processes. However, the mechanisms of action of adiponectin on the maturation of goat oocytes remain to be determined. The aim of this study was to investigate whether (a) adiponectin influences the meiotic maturation of goat oocytes; (b) MAPK MEK 1/2 mediates the effects of adiponectin; and 3) adiponectin differentially affects mRNA relative abundance of genes relevant for adiponectin signal transduction in goat oocytes. The addition of adiponectin (5 µg/ml) during the maturation of goat oocytes resulted in a higher percentage of successful nuclear maturation compared to those of the group without adiponectin (p < 0.05). Adiponectin-stimulated nuclear oocyte maturation was significantly impaired by a mitogen-activated protein kinase MEK 1/2 inhibitor, U0126 (p < 0.05). There was no evidence of any adiponectin-induced difference in the relative transcript abundances of AdipoR1, AdipoR2, AMPKα1, AMPKα2, PPARα and PPARγ genes. In conclusion, these results indicate that adiponectin has a positive effect on the meiotic maturation of goat oocytes through the MAPK MEK 1/2 pathway. Furthermore, the adiponectin does not affect the relative abundance of genes relevant for adiponectin signal transduction in goat oocytes.

Expression of adiponectin and its receptors (AdipoR1 and AdipoR2) in goat ovary and its effect on oocyte nuclear maturation in vitro.

Adiponectin is an adipokine secreted primarily by adipocytes and is involved in the control of male and female reproductive functions. Circulating levels of adiponectin are inversely correlated with body fat mass, and its biological effects are predominantly mediated through two receptors, AdipoR1 and AdipoR2. The aim of the present study was to verify the expression of the adiponectin system (adiponectin and its receptors, AdipoR1 and AdipoR2) in goat ovary using qPCR and immunohistochemistry analyses and further investigate the in vitro effects of recombinant adiponectin (5 µg/mL and 10 µg/mL) on goat oocyte nuclear maturation. We demonstrated that the mRNA and proteins of the adiponectin system are present in goat ovary. Gene and protein expression of AdipoR1 and AdipoR2 was detected in follicular cells (oocyte, cumulus, granulosa and theca) of small and large antral follicles, while adiponectin mRNA was not detected in oocytes from small and large follicles or in large follicle cumulus cells. Finally, addition of various concentrations of adiponectin in maturation medium affected the number of oocytes that reached metaphase II. In conclusion, in the present study, we detected expression of adiponectin and its receptors AdipoR1 and AdipoR2 in goat ovarian follicles. Furthermore, we demonstrated that recombinant adiponectin increases nuclear maturation of goat oocytes in vitro.

Oral administration of an anti-inflammatory does not compromise the efficacy of intra-testicular injection of zinc gluconate as a contraceptive for dogs.

The objective was to determine whether the efficacy of zinc gluconate (Testoblock(®)) as a chemical contraceptive in male dogs was compromised in the presence of metamizole sodium (a nonsteroidal antiinflammatory/analgesic agent). Ten sexually mature mongrel dogs were assigned to two groups, a control group (n = 5) and a treated group (n = 5). Testoblock(®), a proprietary zinc-gluconate (13.1 mg zinc/ml) solution in a physiological vehicle, was injected into each testis (0.2-1.0 ml/testis, based on testis width). Half of the dogs (treated group) were also given metamizole sodium (also known as sodium dipyrone) orally (25mg/kg three times a day for 2 days), starting 2-3 h after testis injection. A physical examination and assessment of testis width, hematology, clinical chemistry (hepatic and renal function) and semen characteristics, were done immediately after treatment and then every 2 months for 180 days. There was no post-treatment scrotal biting or licking, although there was transient testicular swelling in both control and treated dogs during the first 3 days after injection. At 60 days after injection, all dogs were azoospermic. At 120 and 180 days, seven dogs had azoospermia and the remaining three (two control and one treated) had apparent aspermia (no ejaculate could be collected). There were no significant differences between groups for clinical findings or any aspect of hematology, renal, or hepatic function. In conclusion, giving metamizole sodium concurrent with an intra-testicular injection of a zinc-based solution did not interfere with chemical sterilization and it improved animal welfare.

Prevalência e fatores de risco associados à infecção por Toxoplasma gondii em granjas suinícolas tecnificadas no Estado de Alagoas / Prevalence and risk factors associated with infection by Toxoplasma gondii on commercial swine farms in the state of Alagoas, Brazil

Objetivou-se com este estudo calcular a prevalência e identificar os fatores de risco associados à infecção por Toxoplasma gondii em suínos criados em granjas tecnificadas no Estado de Alagoas, Brasil. Para compor a amostra do estudo de prevalência foram utilizados 342 suínos, sendo 312 matrizes e 30 varrões, oriundos de sete granjas de ciclo completo e distribuídas em cinco municípios do Estado de Alagoas. O exame sorológico para a pesquisa de anticorpos anti-Toxoplasma gondii foi realizado através da técnica de Imunofluorescência Indireta (RIFI), utilizando-se anticorpos anti-IgG-suíno conjugado ao isotiocianato de fluoresceína. A análise dos fatores de risco foi realizada por meio da aplicação de questionários constituídos por perguntas objetivas referentes ao criador, às características gerais da propriedade, ao manejo produtivo, reprodutivo e sanitário. Determinou-se uma prevalência de 26,9 por cento (92/342) de suínos soropositivos. O fator associado à infecção foi a introdução de reprodutores nas granjas nos últimos cinco anos (p=0,014; OR=1,83; IC=1,13-2,96). Concluiu-se que a infecção por Toxoplasma gondii encontra-se disseminada em suínos criados em granjas tecnificadas no Estado de Alagoas, Brasil. Recomenda-se o controle da população de gatos, a realização de práticas de limpeza e higienização das instalações e realização de testes laboratoriais para diagnóstico da infecção por T. gondii nos animais a serem introduzidos no plantel como medidas de redução dos índices de infecção na região estudada.

This study was aimed to determine the prevalence and to identify risk factors associated with infection by Toxoplasma gondii in pigs from commercial swine farms in the state of Alagoas, Brazil. To compose the sample size of prevalence 342 pigs were used, with 312 sows and 30 boars, from of seven swine farms and distributed in five districts of the state of Alagoas. The serological examination for the detection of anti-Toxoplasma gondii antibodies was achieved by Indirect Immunofluorescence (IFAT), using anti-pig IgG-conjugated to fluorescein isothiocyanate. The analysis of risk factors were performed by the application of questionnaires consisting of objective questions relating to the designer, the general characteristics of the property, the production, reproductive and health management. The prevalence was of 26,9 percent (92/342) of seropositive pigs. The associated factor was the boars introduction on farms in the last five years (p=0,014; OR=1,83; CI=1,13-2,96). It is concluded that infection with Toxoplasma gondii is widespread in commercial pig farms in the state of Alagoas, Brazil. The results suggest the control of cat population, the performance of cleaning and sanitation practices and laboratory testing to detect of infection by T. gondii in animals to be placed on the farms as measures to reduce the infection rates on region studied.