Canine sperm vitrification with nonpermeable cryoprotectants and coconut water extender

This study was aimed to assess the efficiency of coconut water extender with addition of soy lecithin and sucrose as nonpermeable cryoprotectants for canine semen vitrification, using a simple method that yields a high survival rate of spermatozoa for clinical use. Twelve ejaculates from 12 adult normozoospermic dogs were collected separately by digital manipulation and only the second semen fraction was used in this study. After evaluation of volume, concentration, viability, total and progressive motility, velocity parameters and morphology, semen was diluted with a coconut water extender (50% (v/v(volume per volume)) coconut water, 25% (v/v) distilled water and 25% (v/v) 5% anhydrous monosodium citrate solution) with addition of soy lecithin and fructose at 1% and 0.25M sucrose until final concentration of 100x106 spermatozoa/ml. After equilibration at 5ºC for 60 minutes, semen was vitrified by "direct dropping method" into liquid nitrogen in spheres with a volume of 30 µl. After a week of storage the spheres were devitrified as three of them were dropped into 0.5 mL of CaniPlus AI medium (Minitüb, Germany), which was previously warmed in a water bath at 42ºC for 2 minutes and evaluated about the above mentioned parameters. It was found that vitrification resulted in a lower percentage of viable sperms, normal morphology, total and progressive motilities (p0.05) compared to fresh semen samples. In conclusion, our results demonstrate that vitrification with coconut water extender with addition of 1% soy lecithin and 0.25M sucrose as cryoprotectants, has an excellent potential for routine canine sperm cryopreservation.(AU)

Sustained deficit irrigation on yield and fruit water quality of dwarf green coconut / Efeito da irrigação com déficit hídrico sustentado sobre a produtividade e qualidade da água dos frutos do coqueiro anão verde

This study evaluated the effects of sustained deficit irrigation (SDI) on fruit yield and fruit water quality of dwarf green coconut trees. The experiment was carried out in a commercial orchard located in Camocim, Ceará, Brazil. Four years old coconut trees were irrigated during 29 months, using micro-sprinklers, at irrigation depths equivalent to 55%, 77%, 100% and 131% of crop evapotranspiration (ETc). Green coconut fruits were harvested six months after flower aperture and evaluated for number of fruits per plant, volume of coconut water per fruit and total soluble solids of the coconut water. SDI reduced coconut fruit yield, fruit water volume and coconut water yield. Conversely, SDI increased total soluble solids of the coconut water and irrigation water productivity in terms of fruits and coconut water. Deficit irrigation showed no economic advantage over full irrigation due to the small reduction in irrigation costs compared to the substantial reduction in gross revenue.

Este estudo teve como objetivo avaliar os efeitos da irrigação com déficit sustentado (SDI) sobre a produção e a qualidade da água de frutos do coqueiro Anão Verde. O experimento foi realizado em um pomar comercial localizado em Camocim, Ceará, Brasil. Plantas com quatro anos de idade foram irrigadas por microaspersão, durante 29 meses, com lâminas de irrigação equivalentes a 55%, 77%, 100% e 131% da evapotranspiração da cultura (ETc). Os frutos foram colhidos seis meses após a abertura das flores e avaliados quanto ao número de frutos por planta, volume de água de coco por fruto e sólidos solúveis totais da água de coco. A SDI reduziu a produtividade de frutos, o volume de água do fruto e a produção de água de coco por planta. Por outro lado, a SDI aumentou os sólidos solúveis totais da água de coco e a produtividade da água de irrigação em termos de frutas e água de coco. A SDI não apresentou vantagem econômica sobre a irrigação plena devido à pequena redução nos custos de irrigação em comparação à redução substancial da receita bruta.

Assessment of coconut water added to Numida meleagris egg yolk as cryoprotectant for goat semen

Effects were assessed of the dilutants TRIS and ACP - 101c® with the addition of different guinea fowl (Numida meleagris) egg yolk concentrations. Fifteen ejaculates were collected from five goats of the Anglo Nubian breed. The ejaculates were pooled and then divided into 12 groups, two control groups (GC1 TRIS, with 2.5% Gallus gallus domesticus hen egg yolk GOGD), (GC2 Control Group ACP - 101c®, with the addition of 2.5% Gallus gallus domesticus hen egg yolk GOGD) and ten experimental groups (EG), containing TRIS and ACP added with different concentrations of egg yolk from guinea hen (Numida meleagris) (TRIS 2,5% GONM; TRIS 5% GONM; TRIS 10% GONM; TRIS 15% GONM; TRIS 20% GONM; ACP® 2,5% GONM; ACP® 5% GONM; ACP® 10% GONM; ACP® 15% GONM; ACP® 20% GONM). Then cryopreservation was carried out and the samples stored in liquid nitrogen (-196 °C). After seven days, the samples were thawed and assessed for spermatic kinetics, immunofluorescence and sperm morphology. Analysis of GOMN by the CASA system showed that the various parameters were similar to those of GOGD (P>0.05). The membrane integrity, mitochondrial potential and the acrosome were not influenced by the treatment (P>0.05) nor by the dilutant used for cryopreservation (P>0.05). The spermatic morphology was also preserved by the different GOGD and GONM concentrations in the ACP® and TRIS dilutants, with no statistically significant differences (P<0.05). It was concluded that Numida meleagris egg yolk, as external membrane cryoproctant added to the dilutants ACP-101c® and TRIS, improved goat semen quality.(AU)

Parâmetros microscópicos de açúcar de coco comercializados na cidade de São Paulo, Brasil: identidade e qualidade / Microscopic parameters of coconut sugar marketed in the city of São Paulo, Brazil: identity and quality

O açúcar de coco vem se tornando popular no Brasil e no mundo devido às alegações nutricionais de ser um adoçante natural e mais saudável em relação a outros açúcares. Sua produção ocorre de forma artesanal a partir da extração da seiva do floema da inflorescência da palmeira. Este trabalho teve por objetivo analisar microscopicamente a identidade e a presença de matérias estranhas em amostras de açúcar de coco, bem como verificar sua conformidade em relação às legislações sanitárias brasileiras. Foram analisadas 13 amostras comerciais de açúcar de coco da cidade de São Paulo quanto aos ensaios de pesquisa e identificação de elementos histológicos e pesquisa de matérias estranhas. A identificação de elementos histológicos vegetais revelou a presença de amidos morfologicamente semelhantes ao padrão de Triticum sp. (trigo), Manihot sp. (mandioca) e Oryza sativa (arroz), além de amidos alterados. Também foram encontrados elementos histológicos vegetais compatíveis com padrão de Saccharum officinarum (cana-de-açúcar) e de Cocos nucifera (coco). Todas as amostras analisadas continham matérias estranhas com predomínio de fragmentos de insetos em 100% delas. Os resultados do estudo indicam falhas nas Boas Práticas de produção do açúcar de coco e a necessidade da adequação deste produto em relação às normas vigentes.

Coconut sugar has become popular in Brazil and worldwide due to the nutritional claims of being a natural and healthier sweetener compared to other sugars. Its production takes places in an artisanal way from the extraction of phloem sap from the inflorescence of the palm. The aim of this work was to microscopically analyze the identity and the presence of foreign matter in samples of coconut sugar, as well as to verify its compliance with Brazilian health legislation. Thirteen commercial samples of coconut sugar from the city of São Paulo were analyzed for the research and identification of histological elements and foreign matter. The identification of vegetable histological elements revealed the presence of starches morphologically similar to the Triticum sp. (wheat), Manihot sp. (cassava) and Oryza sativa (rice) pattern, besides altered starches. Vegetable histological elements compatible with the Saccharum officinarum (sugar cane) and Cocos nucifera (coconut) pattern were also found. All samples analyzed contained foreign matter with predominance of insect fragments in 100% of the samples. The results of the study indicate flaws in the Good Practices for the production of coconut sugar and the need to adapt this product to current standards.

Coconut Cake in Diets for Quail in the Laying Phase

Currently, agro-industrial by-products have increasingly been used in animal feeding, as they constitute an alternative source of nutrients for the animal diet and a way to simultaneously reduce environmental pollution. The objective of this study was to examine increasing levels of inclusion of coconut cake in Japanese quail diets in the laying phase on their production performance and egg quality. A total of 360 Japanese quails were allotted to eight treatments with nine replicates and eight birds per experimental unit, in a randomized-block design. Five diets were formulated: a diet without inclusion of the by-product; and diets containing 3, 6, 9, and 12% coconut cake. The experiment lasted 63 days, with evaluations occurring at every 21 days. The following variables were analyzed: feed intake, laying rate, feed conversion, egg weight, specific gravity, Haugh unit, yolk, albumen and shell percentage, shell thickness, and shell weight. The treatments elicited a positive linear response from laying rate, whereas feed conversion per egg mass decreased linearly. In terms of egg-quality traits, shell percentage was influenced, increasing linearly. Coconut cake inclusion at 12% in the diet of Japanese quail in the laying improved feed conversion per egg mass and increased egg-laying rate and eggshell percentage.(AU)

Parâmetros microscópicos de açúcar de coco comercializados na cidade de São Paulo, Brasil: identidade e qualidade / Microscopic parameters of coconut sugar marketed in the city of São Paulo, Brazil: identity and quality

O açúcar de coco vem se tornando popular no Brasil e no mundo devido às alegações nutricionais de ser um adoçante natural e mais saudável em relação a outros açúcares. Sua produção ocorre de forma artesanal a partir da extração da seiva do floema da inflorescência da palmeira. Este trabalho teve por objetivo analisar microscopicamente a identidade e a presença de matérias estranhas em amostras de açúcar de coco, bem como verificar sua conformidade em relação às legislações sanitárias brasileiras. Foram analisadas 13 amostras comerciais de açúcar de coco da cidade de São Paulo quanto aos ensaios de pesquisa e identificação de elementos histológicos e pesquisa de matérias estranhas. A identificação de elementos histológicos vegetais revelou a presença de amidos morfologicamente semelhantes ao padrão de Triticum sp. (trigo), Manihot sp. (mandioca) e Oryza sativa (arroz), além de amidos alterados. Também foram encontrados elementos histológicos vegetais compatíveis com padrão de Saccharum officinarum (cana-de-açúcar) e de Cocos nucifera (coco). Todas as amostras analisadas continham matérias estranhas com predomínio de fragmentos de insetos em 100% delas. Os resultados do estudo indicam falhas nas Boas Práticas de produção do açúcar de coco e a necessidade da adequação deste produto em relação às normas vigentes.(AU)

Coconut sugar has become popular in Brazil and worldwide due to the nutritional claims of being a natural and healthier sweetener compared to other sugars. Its production takes places in an artisanal way from the extraction of phloem sap from the inflorescence of the palm. The aim of this work was to microscopically analyze the identity and the presence of foreign matter in samples of coconut sugar, as well as to verify its compliance with Brazilian health legislation. Thirteen commercial samples of coconut sugar from the city of São Paulo were analyzed for the research and identification of histological elements and foreign matter. The identification of vegetable histological elements revealed the presence of starches morphologically similar to the Triticum sp. (wheat), Manihot sp. (cassava) and Oryza sativa (rice) pattern, besides altered starches. Vegetable histological elements compatible with the Saccharum officinarum (sugar cane) and Cocos nucifera (coconut) pattern were also found. All samples analyzed contained foreign matter with predominance of insect fragments in 100% of the samples. The results of the study indicate flaws in the Good Practices for the production of coconut sugar and the need to adapt this product to current standards.(AU)

Coconut Cake in Diets for Quail in the Laying Phase

Currently, agro-industrial by-products have increasingly been used in animal feeding, as they constitute an alternative source of nutrients for the animal diet and a way to simultaneously reduce environmental pollution. The objective of this study was to examine increasing levels of inclusion of coconut cake in Japanese quail diets in the laying phase on their production performance and egg quality. A total of 360 Japanese quails were allotted to eight treatments with nine replicates and eight birds per experimental unit, in a randomized-block design. Five diets were formulated: a diet without inclusion of the by-product; and diets containing 3, 6, 9, and 12% coconut cake. The experiment lasted 63 days, with evaluations occurring at every 21 days. The following variables were analyzed: feed intake, laying rate, feed conversion, egg weight, specific gravity, Haugh unit, yolk, albumen and shell percentage, shell thickness, and shell weight. The treatments elicited a positive linear response from laying rate, whereas feed conversion per egg mass decreased linearly. In terms of egg-quality traits, shell percentage was influenced, increasing linearly. Coconut cake inclusion at 12% in the diet of Japanese quail in the laying improved feed conversion per egg mass and increased egg-laying rate and eggshell percentage.

Aspectos econômicos e avaliação do período de viabilidade de espermatozoides ovino imunossexados e diluídos em meio de conservação à base de água de coco em pó (ACP-102c) / Economic aspects and evaluation of the viability period of immunosexed and diluted ram sperm in powdered coconut water conservation extender (ACP-102c)

Background: Sperm sexing is increasing in use because pre-determining the sex of the calf allows greater profitability and promotes significant gains in the productive systems that utilize the technique. Deployment of a low-cost and practical preservation methodol-ogy may further favor the cost-benefit ratio. Flow cytometry, the most commonly used sexing technique, has high costs and is very restricted. As an alternative, immunosexing has been studied, which uses sex-specific monoclonal antibodies. Thus, the objective of this study was to evaluate the immunosexing technique in conjunction with cryopreservation in ACP-102c and examine its economic aspects with regard to ram semen.Materials, Methods & Results: Ejaculates from two ram individuals were collected with the aid of an artificial vagina, evaluated, and submitted to the immunosexing protocol, according to the manufacturer’s recommendations, using the Monoclonal Antibody Kit specific for mammalian sperm with “Y” chromosomes (HY; HY Biotechnology, Rio de Janeiro-RJ, Brazil). After sexing, the supernatant was resuspended in the cryopreservation diluent: ACP (ACP-102c + 20% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated to 4°C, stabilized for 30 min, frozen in liquid nitrogen vapors (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were thawed and evaluated for sperm kinetics both by using computerized semen analysis with SCA® software (Sperm Class Analyzer version 5.0) and subjectively comparing specimens from the two animals using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique...

Biotecnologias como ferramentas para o desenvolvimento do Nordeste do Brasil / Biotechnologies as tools for the development of northeast Brazil

A biotecnologia tem sido um ramo de estudo diferencial para diversos setores da sociedade, apresentando, através de bioprodutos e bioprocessos, melhorias para o avanço e desenvolvimento da região Nordeste do Brasil. Vale pontuar que um importante meio que traz anualmente acréscimos inovadores à área biotecnológica é o setor acadêmico, que através de cursos stricto sensu a nível de mestrado e doutorado, promovem pesquisas relevantes para vários setores como economia, agroindústria, saúde, dentre outros. Exemplos disso são: o curso de Doutorado em Biotecnologia da RENORBIO) e o Programa Profissional de Pós-Graduação em Biotecnologia em Saúde Humana e Animal (PPGBiotec). O presente artigo se sobre o percurso trilhado pelos cursos stricto sensu mencionados, bem como ressalta a relevância da Biotecnologia para a região Nordeste do Brasil, em seus diferentes campos de investigação, com ênfase nos Recursos Naturais, Agropecuária e Saúde. Exemplificamos as biotecnologias utilizando a água de coco que vêm sendo trabalhadas desde os anos 1980’s e sua evolução até os dias atuais. Com base em toda a potencialidade da Região Nordeste para a geração de bioprodutos e bioprocessos, ressaltamos que os mesmos só serão úteis se realmente forem tratados como inovação tecnológica, gerarem nota fiscal e impactarem positivamente para o bem estar da sociedade.

Biotechnology has been a branch of differential study for various sectors of society, presenting, through bioproducts and bioprocesses, improvements for the advancement and development of the Northeast region of Brazil. It is worth noting that an important means that annually brings innovative additions to the biotechnological area is the academic sector, which through stricto sensu courses at the master's and doctoral level, promote relevant research for various sectors such as economics, agribusiness, health, among others. Examples of this are: the Doctorate course in Biotechnology (RENORBIO) and the Professional Graduate Program in Biotechnology in Human and Animal Health (PPGBiotec). This article is about the path taken by the stricto sensu courses mentioned, as well as emphasizing the relevance of Biotechnology for the Northeast region of Brazil, in its different research fields, with emphasis on Natural Resources, Agriculture and Health. We exemplify biotechnologies using coconut water that has been worked since the 1980's and its evolution to the present day. Based on all the potential of the Northeast Region for the generation of bioproducts and bioprocesses, we emphasize that they will only be useful if they are really treated as technological innovation, generate invoices and have a positive impact on society's well-being.

Biotecnologias como ferramentas para o desenvolvimento do Nordeste do Brasil / Biotechnologies as tools for the development of northeast Brazil

A biotecnologia tem sido um ramo de estudo diferencial para diversos setores da sociedade, apresentando, através de bioprodutos e bioprocessos, melhorias para o avanço e desenvolvimento da região Nordeste do Brasil. Vale pontuar que um importante meio que traz anualmente acréscimos inovadores à área biotecnológica é o setor acadêmico, que através de cursos stricto sensu a nível de mestrado e doutorado, promovem pesquisas relevantes para vários setores como economia, agroindústria, saúde, dentre outros. Exemplos disso são: o curso de Doutorado em Biotecnologia da RENORBIO) e o Programa Profissional de Pós-Graduação em Biotecnologia em Saúde Humana e Animal (PPGBiotec). O presente artigo se sobre o percurso trilhado pelos cursos stricto sensu mencionados, bem como ressalta a relevância da Biotecnologia para a região Nordeste do Brasil, em seus diferentes campos de investigação, com ênfase nos Recursos Naturais, Agropecuária e Saúde. Exemplificamos as biotecnologias utilizando a água de coco que vêm sendo trabalhadas desde os anos 1980s e sua evolução até os dias atuais. Com base em toda a potencialidade da Região Nordeste para a geração de bioprodutos e bioprocessos, ressaltamos que os mesmos só serão úteis se realmente forem tratados como inovação tecnológica, gerarem nota fiscal e impactarem positivamente para o bem estar da sociedade.(AU)

Biotechnology has been a branch of differential study for various sectors of society, presenting, through bioproducts and bioprocesses, improvements for the advancement and development of the Northeast region of Brazil. It is worth noting that an important means that annually brings innovative additions to the biotechnological area is the academic sector, which through stricto sensu courses at the master's and doctoral level, promote relevant research for various sectors such as economics, agribusiness, health, among others. Examples of this are: the Doctorate course in Biotechnology (RENORBIO) and the Professional Graduate Program in Biotechnology in Human and Animal Health (PPGBiotec). This article is about the path taken by the stricto sensu courses mentioned, as well as emphasizing the relevance of Biotechnology for the Northeast region of Brazil, in its different research fields, with emphasis on Natural Resources, Agriculture and Health. We exemplify biotechnologies using coconut water that has been worked since the 1980's and its evolution to the present day. Based on all the potential of the Northeast Region for the generation of bioproducts and bioprocesses, we emphasize that they will only be useful if they are really treated as technological innovation, generate invoices and have a positive impact on society's well-being.(AU)

Produção in vivo de embriões caprinos e ovinos submetidos ao meio de manutenção ACP® / In vivo production of goat and sheep embryos submitted to the ACP® maintenance médium

The use of ACP® as a means of maintenance in the stages of multiple ovulation and embryo transfer, has not yet been reported in the literature, although ACP® is a plant fluid rich in nutrients. Thus, the objective was to compare the efficiency of ACP® as a substitute for embryo maintenance medium during MOTE biotechnology in goats and sheep. For this, three donor goats, Anglo-Nubian breed and three donor sheep, Dorper breed were used. Fifteen recipient females of each species were used. The donors were submitted to the superovulation protocol and inseminated, and later the embryos were collected. After harvesting, the embryos were submitted to the control maintenance medium, TQC Holding® or ACP® maintenance medium. The recipients were synchronized simultaneously with the donors, and after 30 days the pregnancy diagnosis was made. It was obtained 10% of pregnant in goats and 75% of pregnant in sheep, whose embryos were submitted to the ACP® maintenance medium before the innovation. It is concluded that the means of maintenance of ACP® embryos did not negatively influence the embryonic quality and the development of pregnancy in small ruminants.

Produção in vivo de embriões caprinos e ovinos submetidos ao meio de manutenção ACP® / In vivo production of goat and sheep embryos submitted to the ACP® maintenance médium

The use of ACP® as a means of maintenance in the stages of multiple ovulation and embryo transfer, has not yet been reported in the literature, although ACP® is a plant fluid rich in nutrients. Thus, the objective was to compare the efficiency of ACP® as a substitute for embryo maintenance medium during MOTE biotechnology in goats and sheep. For this, three donor goats, Anglo-Nubian breed and three donor sheep, Dorper breed were used. Fifteen recipient females of each species were used. The donors were submitted to the superovulation protocol and inseminated, and later the embryos were collected. After harvesting, the embryos were submitted to the control maintenance medium, TQC Holding® or ACP® maintenance medium. The recipients were synchronized simultaneously with the donors, and after 30 days the pregnancy diagnosis was made. It was obtained 10% of pregnant in goats and 75% of pregnant in sheep, whose embryos were submitted to the ACP® maintenance medium before the innovation. It is concluded that the means of maintenance of ACP® embryos did not negatively influence the embryonic quality and the development of pregnancy in small ruminants.(AU)

Avaliação microbiológica de coco ralado úmido e adoçado obtido no comércio / Microbiological evaluation of damp and sweetened coconut flakes obtained in trade

Coco ralado é o produto obtido do fruto do coqueiro (Cocos nucifera L.), podendo ser classificado quanto á umidade, adição de açúcar e caramelização. Por ser facilmente encontrado em comércios e amplamente utilizado, o objetivo desse trabalho foi avaliar a qualidade microbiológica de coco ralado úmido e adoçado comercializados nos estados do Rio de Janeiro e Mato Grosso. Das três amostras analisadas, apenas uma apresentou coliformes a 45° enquanto que duas apresentaram Staphylococcus coagulase positiva acima do limite estabelecido em legislação, além disso, duas das três amostras apresentaram suspeita de presença de Salmonella sp. De acordo com os resultados obtidos, duas amostras estavam impróprias para o consumo, pois estavam fora dos padrões exigidos pela legislação brasileira.(AU)

Avaliação microbiológica de coco ralado úmido e adoçado obtido no comércio / Microbiological evaluation of damp and sweetened coconut flakes obtained in trade

Coco ralado é o produto obtido do fruto do coqueiro (Cocos nucifera L.), podendo ser classificado quanto á umidade, adição de açúcar e caramelização. Por ser facilmente encontrado em comércios e amplamente utilizado, o objetivo desse trabalho foi avaliar a qualidade microbiológica de coco ralado úmido e adoçado comercializados nos estados do Rio de Janeiro e Mato Grosso. Das três amostras analisadas, apenas uma apresentou coliformes a 45° enquanto que duas apresentaram Staphylococcus coagulase positiva acima do limite estabelecido em legislação, além disso, duas das três amostras apresentaram suspeita de presença de Salmonella sp. De acordo com os resultados obtidos, duas amostras estavam impróprias para o consumo, pois estavam fora dos padrões exigidos pela legislação brasileira.

Supplemented powdered coconut water (ACP-406®) promotes growth of goat secondary follicles and oocyte meiotic resumption

The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.(AU)

Supplemented powdered coconut water (ACP-406®) promotes growth of goat secondary follicles and oocyte meiotic resumption

The objective of this study was to test the efficiency of powdered coconut water (ACP-406®) base-medium without or with the addition of supplements on in vitro culture of isolated goat secondary follicles. Follicles were cultured for 18 days in α-MEM or in ACP-406®, both without supplements (referred to as α-MEM and ACP, respectively), or both supplemented with BSA, insulin, transferrin, selenium, glutamine, hypoxanthine, and ascorbic acid (referred to as α-MEM+ and ACP+). Follicular morphology, antrum formation, follicular and oocyte diameter, levels of glutathione (GSH), and chromatin configuration after in vitro maturation were evaluated. At the end of culture, ACP-406® base-medium (without or with supplements) showed a higher (P < 0.05) percentage of normal follicles than α-MEM (without or with supplements). Antrum formation was similar among α-MEM+, ACP and ACP+, and significantly higher than α-MEM without supplements. The follicular diameter was greater in ACP+ than α-MEM, and similar to other treatments. Moreover, fully and daily grown rates were higher (P < 0.05) in ACP-406® base-medium (without or with supplements) than α-MEM (without or with supplements). Levels of GSH were similar between ACP+ and α-MEM+ treatments. Both ACP+ and α-MEM+ allowed meiotic resumption without a significant difference between the two groups. In conclusion, supplemented ACP-406® base-medium maintained follicular survival and promoted the development as well as meiotic resumption of isolated goat secondary follicles cultured in vitro for 18 days.

Influência do meio de conservação à base de água de coco em pó (ACP-102c) na manutenção da atividade mitocondrial de espermatozoides ovinos criopreservados / Influence of coconut powder water-based conservation medium (APC-102c) for maintaining mitochondrial activity of cryopreserved ram sperm

Background: Semen extenders are required to protect and preserve semen, and the development of suitable extenders iskey for artificial insemination. Although the use of Tris-based diluent is widespread, new diluents such as powdered coconut water have been developed for better sperm protection. One way to evaluate the effectiveness of diluents is throughmicroscopic analyses that evaluate sperm motility, vigor, and concentration. However, these analyses are limited, and maynot provide accurate results. New evaluation techniques have been studied, and one of the tests that can be used to addreliability to these analyses is mitochondrial activity evaluation, which can sum all the parameters, and provide a moreaccurate evaluation. Thus, the present study aimed to evaluate the efficacy of ACP-102c in cryopreserved ram semen.Materials, Methods & Results: Five semen samples were collected from two ram breeders using artificial vagina (n = 10).Each ejaculate was divided into the following two treatments: T1 - ACP-102c + 20% egg yolk + 7% glycerol and T2 - TRIS+ 20% egg yolk + 7% glycerol. Extended semen samples were then packed in 0.5 mL plastic straws, subjected through therefrigeration curve up to 4°C (0.35°C/min), and equilibrated for 2 h at 4°C. Subsequently, the straws were placed at 4 cmabove liquid nitrogen level (-60°C) for 15 min, immersed, and then finally stored in the liquid nitrogen at -196°C. Bothfresh and thawed samples were evaluated for total and progressive sperm motility using conventional microscopy (40x),and the same evaluator on each occasion. For plasma membrane integrity (IMP), the smear staining technique with theEosin-Nigrosin staining was used; 200 sperms were counted and classified as whole (unstained) and unhealthy (stained).Mitochondrial activity was evaluated using a cytochemical technique based on the oxidation of 3,3-diaminobenzidine(DAB); 200 sperms were counted, and classified into four...(AU)

Powdered coconut water (ACP 406®) as an alternative base culture medium for in vitro culture of goat preantral follicles enclosed in ovarian tissue

This study evaluated a powdered coconut water solution (ACP 406®) as a base culture medium on the in vitro survival and development of in situ goat preantral follicles. The ovarian fragments were either immediately fixed in Carnoy solution (non-cultured control) or individually cultured for 2 or 6 days. The following culture media (all containing 100 μg/mL penicillin and 100 μg/mL streptomycin) were evaluated: α-MEM (α-MEM alone, without additional supplementation); α-MEM+ (supplemented α-MEM); ACP (ACP®406 alone); or ACP+ (supplemented ACP®406). Additional supplementation includes: 1.25 mg/mL bovine serum albumin, 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 2 mM glutamine, and 2 mM hypoxanthine. The endpoints (i) follicular morphology; (ii) development; (iii) estradiol production; and (iv) reactive oxygen species (ROS) were recorded. Data were analyzed using chi-square, Turkey, t-test or One-Way ANOVA. Differences were considered significant when P < 0.05. At day 2 of culture, a greater (P < 0.05) percentage of morphologically normal follicles was observed between ACP+ and ACP treatments. Moreover, at day 2 of culture, no hormonal difference (P < 0.05) was observed between ACP+ and both α-MEM treatments. At day 6 of culture when ACP and α-MEM treatments were compared the percentage of healthy follicles were similar (P > 0.05) among treatments. Overall, all treatments had lower primordial follicles (P < 0.05) accompany by greater developing follicles (P < 0.05) percentages than non-cultured control treatment, indicating primordial follicle activation. However, at day 6 of culture, the percentage of primordial follicle development were similar (P > 0.05) among the treatments. Likewise, no differences (P > 0.05) were observed for ROS production and follicular and oocyte diameters among treatments. Therefore, ACP+ has the equivalent efficiency to MEM+ in maintaining the survival and development of goat preantral follicles, representing an alternative plant-based low-cost culture medium for in vitro culture.(AU)

Powdered coconut water (ACP 406®) as an alternative base culture medium for in vitro culture of goat preantral follicles enclosed in ovarian tissue

This study evaluated a powdered coconut water solution (ACP 406®) as a base culture medium on the in vitro survival and development of in situ goat preantral follicles. The ovarian fragments were either immediately fixed in Carnoy solution (non-cultured control) or individually cultured for 2 or 6 days. The following culture media (all containing 100 μg/mL penicillin and 100 μg/mL streptomycin) were evaluated: α-MEM (α-MEM alone, without additional supplementation); α-MEM+ (supplemented α-MEM); ACP (ACP®406 alone); or ACP+ (supplemented ACP®406). Additional supplementation includes: 1.25 mg/mL bovine serum albumin, 10 μg/mL insulin, 5.5 μg/mL transferrin, 5 ng/mL selenium, 2 mM glutamine, and 2 mM hypoxanthine. The endpoints (i) follicular morphology; (ii) development; (iii) estradiol production; and (iv) reactive oxygen species (ROS) were recorded. Data were analyzed using chi-square, Turkey, t-test or One-Way ANOVA. Differences were considered significant when P 0.05) among treatments. Overall, all treatments had lower primordial follicles (P 0.05) among the treatments. Likewise, no differences (P > 0.05) were observed for ROS production and follicular and oocyte diameters among treatments. Therefore, ACP+ has the equivalent efficiency to MEM+ in maintaining the survival and development of goat preantral follicles, representing an alternative plant-based low-cost culture medium for in vitro culture.

Influência do meio de conservação à base de água de coco em pó (ACP-102c) na manutenção da atividade mitocondrial de espermatozoides ovinos criopreservados / Influence of coconut powder water-based conservation medium (APC-102c) for maintaining mitochondrial activity of cryopreserved ram sperm

Background: Semen extenders are required to protect and preserve semen, and the development of suitable extenders iskey for artificial insemination. Although the use of Tris-based diluent is widespread, new diluents such as powdered coconut water have been developed for better sperm protection. One way to evaluate the effectiveness of diluents is throughmicroscopic analyses that evaluate sperm motility, vigor, and concentration. However, these analyses are limited, and maynot provide accurate results. New evaluation techniques have been studied, and one of the tests that can be used to addreliability to these analyses is mitochondrial activity evaluation, which can sum all the parameters, and provide a moreaccurate evaluation. Thus, the present study aimed to evaluate the efficacy of ACP-102c in cryopreserved ram semen.Materials, Methods & Results: Five semen samples were collected from two ram breeders using artificial vagina (n = 10).Each ejaculate was divided into the following two treatments: T1 - ACP-102c + 20% egg yolk + 7% glycerol and T2 - TRIS+ 20% egg yolk + 7% glycerol. Extended semen samples were then packed in 0.5 mL plastic straws, subjected through therefrigeration curve up to 4°C (0.35°C/min), and equilibrated for 2 h at 4°C. Subsequently, the straws were placed at 4 cmabove liquid nitrogen level (-60°C) for 15 min, immersed, and then finally stored in the liquid nitrogen at -196°C. Bothfresh and thawed samples were evaluated for total and progressive sperm motility using conventional microscopy (40x),and the same evaluator on each occasion. For plasma membrane integrity (IMP), the smear staining technique with theEosin-Nigrosin staining was used; 200 sperms were counted and classified as whole (unstained) and unhealthy (stained).Mitochondrial activity was evaluated using a cytochemical technique based on the oxidation of 3,3’-diaminobenzidine(DAB); 200 sperms were counted, and classified into four...