Establishing an in vitro production program for buffalo embryos (Bubalus bubalis) in Colombia

Objective. Evaluate the results of the standardization of the in vitro production program of buffalo embryos, using oocytes obtained by ultrasound guided oocyte puncture during the 2012 breeding season in Colombia. Materials and methods. Fifty seven buffalo females were selected for ultrasound guided transvaginal aspiration of follicles, oocytes were identified within follicular fluid, classified and transported to the laboratory and matured in vitro for 18 to 20 hours. Frozen semen of seven Mediterranean bulls were used, motile sperm was obtained using the Percoll technique and oocytes were inseminated with 2 million sperm/ml. Presumptive zygotes were cultured for 6 days, grade 1 embryos obtained were frozen using ethylene glycol. Embryos were transferred to females on day 5 during natural cycle. Results. 97 aspirations were performed on the 57 animals, in 8.2% of the aspirations no oocytes were found. 8 oocytes/aspiration were obtained. Of the 783 oocytes, 92% were classified as viable (721/783) and were fertilized. The cleavage and blastocyst rate were 23% and 19% respectively. 37 embryos were transferred and 11 pregnancies were obtained, confirmed by rectal palpation 60 days after transfer, with a pregnancy rate of 29.7%. Conclusions. The results presented here are comparable with others in literature and shows the feasibility of producing in vitro embryos and pregnancies after the standardization of current protocols, with normal and sexed semen and transfer during natural cycle in buffalo.Key words: Buffaloes, embryo, fertilization in vitro, reproduction (Source: MeSH)

In vitro proizvodnja zametaka bizona u Kolumbiji - rezultati dodavanja faktora rasta i antioksidansa definiranom mediju za uzgoj zametaka

To increase the success rate of the in vitro production (IVP) system of embryos in buffalo, it is necessary to improve parameters. This field requires more research since its results are low compared to cattle. In this study carried out in Colombia, a modification of the methodology to produce buffalo IVP embryos is described, and 24 non-lactating females, aged 4 to 11 years old were included. Follicular wave was synchronised by ablation of the follicles within the ovary, seven days later all follicles were aspirated to obtain oocytes for maturation, fertilisation and embryo production. For all culture media used, animal protein was avoided. Culture media were supplemented with growth factors and other molecules associated with embryonic development. Special care was taken in adding supplements to protect the zygotes against the harmful effect of oxygen free radicals in the culture medium. Two aspiration series were performed 15 days apart. The average number of oocytes obtained by aspiration was 11.33, the number of blastocysts was 3.05, and the obtained blastocyst production rate (36.2%) was higher than literature reports for this species; we hypothesise that this increase is associated with the supplements added. It was possible to observe a female that produced 50 oocytes. Subsequent research is needed to evaluate the effect of the medium on embryo quality and pregnancy rates. A cost analysis of the new proposed medium is required, since all supplements must be imported and handled with a cold chain of up to -80oC, making it more expensive and restricting widespread use. Finally, it is shown that with three embryos per aspiration, at least one pregnancy can be successful after the aspiration, which is one of the objectives of the IVP system.Za povećanje djelotvornosti sustava in vitro proizvodnje (IVP) zametaka u bizona, potrebno je poboljšati parametre. Budući da su rezultati slabi u usporedbi s govedima slavi ovo područje zahtijeva više istraživanja. U ovom radu, provedenom u Kolumbiji, modifikacija metodologije za proizvodnju IVP zametaka bizona opisana je na sljedeći način: korištene su 24 ženke u dobi između 4 i 11 godina, a koje nisu bile u laktaciji. Folikularni val je sinkroniziran ablacijom folikula unutar jajnika, a nakon sedam dana obavljena je aspiracija svih folikula ženki za dobivanje oocista za dozrijevanje, oplodnju i proizvodnju embrija. Za sve rabljene podloge izbjegavan je životinjski protein. Podloge su obogaćene faktorima rasta i drugim molekulama povezanima s razvojem embrija. Posebna pozornost posvećena je pri dodavanju dodataka za zaštitu zigota od štetnih učinaka slobodnih radikala u podlozi. Obavljene su dvije aspiracijske serije u razmaku od 15 dana. Prosječni je broj oocista dobivenih aspiracijom bio 11,33, broj blastocista bio je 3,05, a dobivena je stopa proizvodnje blastocista (36,2 %) bolja je u usporedbi s onom koju su objavili drugi autori u državi, za istu vrstu. Pretpostavljamo da je ovo povećanje povezano s dodanim dodatcima, bilo je moguće promatrati ženku koja je proizvela 50 oocista. Naknadno je istraživanje potrebno za procjenu učinka podloge na kvalitetu zametaka i postotka gravidnosti; potrebna je i analiza troškova novog predloženog medija, jer se svi dodatci moraju uvesti u Kolumbiju i čuvati u hladnim uvjetima do minus 80 oC, što ih čini skupljima, a ograničava i njihovu široku primjenu. Na kraju našeg istraživanja uočeno je da s tri zametka po aspiraciji u ove vrste nije teško postići najmanje jednu gravidnost, a to je i jedan od ciljeva IVP-a

Determinación de la calidad microbiológica en productos cosméticos comercializados en el mercado de Salamanca de Monterrico – Ate

Actualmente, los límites de riesgo para la salud de los diferentes productos cosméticos comercializados son cada vez más cortos, por lo que el control microbiológico de estos productos es fundamental para respaldar la seguridad de los usuarios y la calidad del producto y conservar una buena imagen de empresas manufactureras o comercializadoras en el mercado. En el actual estudio se realizó la determinación de la calidad microbiológica de 50 muestras de lápices labiales que se comercializan en los mercados de Salamanca de Monterrico – Ate. Los análisis se han realizado de acuerdo con lo que indica la USP 43 y las Normas técnicas peruanas para cosméticos. Como resultados se obtuvo que: se detectó un aumento de hasta 120 ufc/g en el número total de microorganismos aerobios mesófilos, ausencia de Pseudomonas aeruginosa, presencia de Staphylococcus aureus en dos muestras y ausencia de Escherichia col, en el cálculo total de levaduras y mohos todas las muestras presentaron recuento llegando a obtenerrecuentos de MNPC (Muy numeroso para contar). El 100 % de las muestras se encontraron dentro de especificación del análisis del número total de microorganismos aerobios mesófilos totales, sin embargo, el 4 % presentó presencia de patógenos y el 36% presentó recuento fuera de especificación de mohos y levaduras. En conclusión, los lápices labiales comercializados en los mercados de Salamanca de Monterrico – Ate no cumplen con la calidad microbiológica arriesgando la salud de los usuarios finales

Establishing an in vitro production program for buffalo embryos (Bubalus bubalis) in Colombia

Objective. Evaluate the results of the standardization of the in vitro production program of buffalo embryos, using oocytes obtained by ultrasound guided oocyte puncture during the 2012 breeding season in Colombia. Materials and methods. Fifty seven buffalo females were selected for ultrasound guided transvaginal aspiration of follicles, oocytes were identified within follicular fluid, classified and transported to the laboratory and matured in vitro for 18 to 20 hours. Frozen semen of seven Mediterranean bulls were used, motile sperm was obtained using the Percoll technique and oocytes were inseminated with 2 million sperm/ml. Presumptive zygotes were cultured for 6 days, grade 1 embryos obtained were frozen using ethylene glycol. Embryos were transferred to females on day 5 during natural cycle. Results. 97 aspirations were performed on the 57 animals, in 8.2% of the aspirations no oocytes were found. 8 oocytes/aspiration were obtained. Of the 783 oocytes, 92% were classified as viable (721/783) and were fertilized. The cleavage and blastocyst rate were 23% and 19% respectively. 37 embryos were transferred and 11 pregnancies were obtained, confirmed by rectal palpation 60 days after transfer, with a pregnancy rate of 29.7%. Conclusions. The results presented here are comparable with others in literature and shows the feasibility of producing in vitro embryos and pregnancies after the standardization of current protocols, with normal and sexed semen and transfer during natural cycle in buffalo.Objetivo. Evaluar los resultados de la estandarización de la técnica de producción in vitro de embriones de búfalo, a partir de oocitos obtenidos por punción folicular durante la estación reproductiva del 2012 en una hacienda en Cordoba, Colombia. Materiales y métodos. Cincuenta y siete búfalas fueron seleccionadas para aspiración transvaginal de folículos guiada por ultrasonido, los oocitos fueron identificados y madurados in vitro. Se utilizó semen congelado de 7 búfalos de la raza Mediterráneo para la fertilización in vitro. La fracción móvil fue separada en un gradiente de Percoll, los oocitos fueron inseminados con 2 millones de espermatozides/mL, los presuntos cigotos fueron cultivados por 6 días y los embriones grado 1 obtenidos fueron congelados utilizando etilenglicol. Posteriormente, los embriones fueron transferidos el día 5 post estro en búfalas en ciclo natural. Resultados. En las 57 búfalas, se realizaron 97 aspiraciones foliculares, no se obtuvieron oocitos en el 8.2% de los procedimientos. Se obtuvieron en promedio 8 oocitos por búfala en cada sesión de aspiración. Se recolectaron 783 oocitos de los cuales fueron clasificados como viables el 92% (721/783), se obtuvo una taza de clivaje del 23% y de blastocistos del 19%. De 37 embriones transferidos se obtuvieron 11 preñeces, confirmadas por palpación rectal a los 60 días postransferencia, obteniéndose una taza de preñez del 29.7% Conclusiones. Los resultados presentados en este trabajo son comparables con los de la literatura, en la cual se muestra cómo es posible obtener embriones de semen convencional y sexado, además de producir gestaciones con protocolos estándar de fertilización in vitro y transferencia en ciclos con celo natural adaptados para la especie