In vitro embryo production in small ruminants

This paper reviews the technical bases of in vitro embryo production in small ruminants with special attention to the results obtained by our group in Northeastern Brazil. The laparoscopic oocyte recovery in hormonally treated live animals indicates a promising future for the application of this technique to genetic improvement program. New molecular biology tools should provide information to improve the efficiency of in vitro maturation. Furthers efforts have to be made to improve the oocyte maturation and to standardize the semen-capacitating process

Production of recombinant proteins in milk of transgenic and non-transgenic goats

Among all the transgenic mammalians produced so far, goats have represented an excellent model of transgenesis when considering the factors such as the market demand for protein, volume of milk produced per lactation and reproductive rate. Various recombinant proteins have been obtained from the transgenic and non-transgenic goats, and among these, human antithrombin, produced by the transgenic goats, was the first recombinant protein of animal origin to be released as a drug for the clinical use in humans. This review reports the aspects inherent to the production of recombinant proteins in the goats, from the production of the animal bioreactors up to the expression of these proteins in their milk

In vitro production of small ruminant embryos: Late improvements and further research

Beyond the potential use of in vitro production of embryos (IVP) in breeding schemes, embryos are also required for the establishment of new biotechnologies such as cloning and transgenesis. Additionally, the knowledge of oocyte and embryo physiology acquired through IVP techniques may stimulate the further development of other techniques such as marker assisted and genomic selection of preimplantation embryos, and also benefit assisted procreation in human beings. Efficient in vitro embryo production is currently a major objective for livestock industries, including small ruminants. The heterogeneity of oocytes collected from growing follicles by laparoscopic ovum pick up or in ovaries of slaughtered females, remains an enormous challenge for IVM success, and still limits the rate of embryo development. In addition, the lower quality of the IVP embryos, compared with their in vivo-derived counterparts, translates into poor cryosurvival, which restricts the wider use of this promising technology. Therefore, many studies have been reported in an attempt to determine the most suitable conditions for IVM, IVF, and in vitro development to maximize embryo production rate and quality. This review aims to present the current panorama of IVP production in small ruminants, describing important steps for its success, reporting the recent advances and also the main obstacles identified for its improvement and dissemination

Saneamento básico e impactos ambientais na bacia hidrográfica do Rio Curu – Estado do Ceará – ne do Brasil

A bacia hidrográfica do rio Curu, localizada na porção noroeste do estado do Ceará, Região Nordeste do Brasil, vem sendo assistida por ações governamentais (federal e estadual) desde a década de 1930. Destaca-se dentre as outras bacias do Estado por ter sido a primeira a implantar o sistema de perímetros irrigados (1975) e a organizar um Comitê de Bacia Hidrográfica (1996). A presente pesquisa, de caráter inédito, tem por objetivo analisar as condições ambientais das sedes municipais da bacia do Curu e utilizou como principal método a “listagem de controle” (check-list). Durante o estudo foi observado que a infra-estrutura de saneamento básico não acompanhou o desenvolvimento econômico e o crescimento demográfico da região, revelando-se como potencial poluidora dos recursos hídricos superficiais, através do lançamento de efluentes provindos das atividades urbanas e rurais. Isso acarreta vários prejuízos ambientais, como a alteração da qualidade das águas e o aumento da incidência de doenças de veiculação hídrica.Since 1932, Curu hydrographic basin, placed in the northwest of the Ceará state, has been assisted by governmental actions in both federal and state levels. It stands out among other Ceará basins because has been the first to implement the irrigated perimeter system (1975) and also to organise a Basin Committee (1996). Nowadays, it shows serious environmental problems related with the deficient basic sanitation and the effects of inadequate utilization of the natural resources. That way, this investigation, of unpublished character, analyze the sanitary conditions of the districts of this basin, identifying through "check list" method. During this study it was observed that the basic sanitation did not expand together with the economic and demographic development of the region, being a potential pollutant of the hydric superficial recourse. This results in several environmental damage such change of the water quality and the increase of hydric vehicle disease.La cuenca hidrográfica del río Curu, localizada en la porción noroeste del estado de Ceará, región Nordeste de Brasil, está siendo asistida por acciones gubernamentales (federales y estatales) desde la década de 1930. Se ha sobresalido entre las otras bacías del estado por haber sido la primera a implementar el sistema de perímetros irrigados (1975) y a organizar un Comité de Bacía Hidrográfica (1996). Actualmente posee serios problemas ambientales respecto a la deficiente infraestructura de saneamiento básico de las ciudades que componen la bacía y a los efectos de la utilización inadecuada de los recursos naturales. Así que la presente investigación, de carácter inédito, analiza las condiciones ambientales de las sedes municipales de bacía hidrográfica del Curu, considerando la precariedad del saneamiento básico como uno de los principales factores potenciales de degradación de los recursos hídricos superficiales. En visita a campo se identificó los aspectos sanitarios de las sedes municipales, a través del método "listado de control" (check-list). A lo largo del estudio se observó que la infraestructura no acompañó el desarrollo económico y el crecimiento demográfico de la región, revelándose la falta de este tipo de infraestructura como factor potencial de polución de los recursos hídricos superficiales, ya que los efluentes provenientes de actividades urbanas y rurales se tiran al río. Eso trae varios perjuicios ambientales, como la alteración de la calidad de las aguas y el aumento de la incidencia de enfermedades de transmisión por medio hídrico.A bacia hidrográfica do rio Curu, localizada na porção noroeste do estado do Ceará, Região Nordeste do Brasil, vem sendo assistida por ações governamentais (federal e estadual) desde a década de 1930. Destaca-se dentre as outras bacias do Estado por ter sido a primeira a implantar o sistema de perímetros irrigados (1975) e a organizar um Comitê de Bacia Hidrográfica (1996). A presente pesquisa, de caráter inédito, tem por objetivo analisar as condições ambientais das sedes municipais da bacia do Curu e utilizou como principal método a “listagem de controle” (check-list). Durante o estudo foi observado que a infra-estrutura de saneamento básico não acompanhou o desenvolvimento econômico e o crescimento demográfico da região, revelando-se como potencial poluidora dos recursos hídricos superficiais, através do lançamento de efluentes provindos das atividades urbanas e rurais. Isso acarreta vários prejuízos ambientais, como a alteração da qualidade das águas e o aumento da incidência de doenças de veiculação hídrica

Porcine oocyte preincubation in oviductal fluid flush before in vitro fertilization in the presence of oviductal epithelial cells improves monospermic zygote production

The present study was designed to evaluate the effect of the combination of oviduct fluid flush (OFF) and oviduct epithelial cells (OEC) in modulating the incidence of polyspermy in pigs. Therefore, for in vitro fertilization (IVF), oocyte and sperm were co-cultured in Tris-buffered medium (TBM) either supplemented with 10% OFF (OFFD group), or in the presence of a bovine OEC monolayer (OEC group), or the oocytes were exposed to OFF for 30 min before IVF (OFFB group), or in the presence of an OEC monolayer (OFFB + OEC group). Regardless of sperm concentration used (0.5, 1.5, and 4.5 × 105 cells/ml), supplementation of IVF medium with 10% OFF led to an increased (P 0.05) of the penetration rate in comparison with the control and OEC groups. When the IVF medium was supplemented with heparin, an overall increase (P < 0.05) of the final output of the IVF system in terms of zygotes with two pronuclei (2PN) was observed in the OFFD group, compared with the control and OEC groups, at a sperm concentration of 4.5 × 105 cells/ml. At this concentration, OFFB improved the monospermy rate but decreased the penetration rate, resulting in low efficiency of monospermic zygotes production. Despite this, no major effect was observed in the developmental competence of the presumed zygotes up to the blastocyst stage. The combination of OFFB with OEC improved the penetration rate, while maintaining the high monospermic rate induced by OFFB. In conclusion, the combination of treatment of oocytes by diluted OFF 30 min before IVF, followed by IVF in the presence of OEC, improved monospermic zygote production without reducing the penetration rate, when the IVF medium was supplemented with heparin.Fil: Batista, Ribrio Ivan Tavares Pereira. Institut National de la Recherche Agronomique; FranciaFil: Moro, Lucía Natalia. Institut National de la Recherche Agronomique; Francia. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Corbin, Emilie. Institut National de la Recherche Agronomique; FranciaFil: Alminana, Carmen. Institut National de la Recherche Agronomique; FranciaFil: Goncalves de Souza Fabjan, Joanna Maria. Universidade Federal do Ceara; BrasilFil: de Figueirêdo Freitas, Vicente José. Universidade Federal do Ceara; BrasilFil: Mermillod, Pascal. Institut National de la Recherche Agronomique; Franci

Combination of oviduct fluid and heparin to improve monospermic zygotes production during porcine in vitro fertilization

In vivo, the oviduct provides appropriate microenvironment conditions for monospermic fertilization and early embryo development. In addition, glycosaminoglycans such as heparin are present in the oviduct and have been shown to modulate the activity of oviduct-secreted proteins on the regulation of sperms parameters. Thus, the present study was designed to evaluate the effect of porcine oocytes exposure to oviduct fluid (OF) before in vitro fertilization (IVF; incubation of oocytes in OF for 30 minutes before IVF), during IVF (supplementation of IVF medium with 10% OF), and during IVF in combination with heparin (10% OF + 10-μg/mL heparin) on IVF parameters. Regardless of sperm concentration used (0.5, 1.5, or 4.5 × 105 cells/mL), exposure of oocytes to OF led to an increased (P 0.05) of the penetration rate in comparison with the control group. This resulted in a general increase (P 0.05) on the monospermy rate in comparison with 10% OF alone. This resulted in a general reduction (P < 0.05) in the final output of the IVF system (%), which was 33 ± 6% and 52 ± 8%, for 10% OF + heparin and 10% OF, respectively. In conclusion, the OF, used in porcine IVF, exerted a beneficial effect on oocytes by reducing the incidence of polyspermy without decreasing the penetration rate. However, the association of the OF with heparin reduced the efficiency of monospermic zygotes' production.Fil: Tavares Pereira Batista, Ribrio Ivan. Centre National de la Recherche Scientifique; Francia. Universidade Estadual Do Ceara; BrasilFil: Moro, Lucía Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Centre National de la Recherche Scientifique; FranciaFil: Corbin, Emilie. Centre National de la Recherche Scientifique; FranciaFil: Alminana, Carmen. Centre National de la Recherche Scientifique; FranciaFil: Gonçalves Souza Fabjan, Joanna Maria. Universidade Estadual Do Ceara; BrasilFil: de Figueirêdo Freitas, Vicente José. Universidade Estadual Do Ceara; BrasilFil: Mermillod, Pascal. Centre National de la Recherche Scientifique; Franci