BIOTECNOLOGIAS COMO FERRAMENTAS PARA O DESENVOLVIMENTO DO NORDESTE DO BRASIL

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.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

Criopreservação do sêmen de caprinos em diluidores alternativos e análise da viabilidade / Cryopreservation of goat semen in alternative extenders and viability analysis

Esse trabalho teve como objetivo avaliar os efeitos da refrigeração e congelação do sêmen caprino utilizando diluentes de origem vegetal. Para isso, foi utilizado 20 amostras de sêmen de cinco machos caprinos, em que foi avaliado imediatamente após a coleta, diluídos em ACP-101 e extrato liofilizado de Aloe vera e controle Tris. O sêmen foi refrigerado a 4 °C e congelado a -196 °C. Ao término da refrigeração e após descongelação as amostras foram avaliadas quanto aos parâmetros cinéticos de motilidade, velocidades e viabilidade. Os resultados das avaliações, demonstraram que o diluente ACP apresentou resultados superiores em comparação aos tratamentos com Aloe vera. Portanto, a utilização do ACP-101 associado aos crioprotetores proporciona um excelente meio para criopreservação do sêmen de caprinos. Novos estudos são necessários para verificar em que concentração o aloe vera pode ser utilizado na criopreservação do semen exercendo outros beneficios

Ram and Goat Semen Immunosexed and Diluted in Powdered Coconut Water-Based Preservation Medium (ACP101/102c).

Background: Sperm sexing aims to separate sperm populations in carriers of the “X” or “Y” chromosome. Currently, flow cytometry is a technique that allows greater accuracy; however, it causes structural changes in sperm, reduces viability, and has a high cost. As a result, other methods have been researched, including immunosexing, which uses monoclonal antibodies to detect sex-specific surface antigens. Thus, the objective of this study was to evaluate the immunosexing technique using a monoclonal antibody against sex-specific protein (HY) in the conservation of ram and goat semen in ACP101/102c.Materials, Methods & Results: Ejaculates from 5 rams and 5 goats were collected with the aid of an artificial vagina; they were 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 ram (ACP101/102c + 20% egg yolk + 7% glycerol) and ACP goat (ACP101/102c + 2.5% egg yolk + 7% glycerol), packaged in 0.25 mL straws, refrigerated at 4°C, stabilized for 30 min, frozen in liquid nitrogen vapor (-60°C) for 15 min, immersed in liquid nitrogen, and stored in cryogenic cylinders. The samples were evaluated in natura (T1), after immunosexing (T2) and after thawing (T3) for sperm motility subjectively using conventional microscopy (40x). Plasma membrane integrity (IMP) and sperm cell morphology were evaluated by the smear staining technique using eosin-nigrosine dye, and the percentages of healthy and morphologically defect spermatozoa were determined. In the evaluation of ram semen regarding sperm motility and IMP, no statistically significant differences were observed between treatments after sexing in the evaluation of absolute data (P > 0.05), with the difference being observed only between T1 and T2, and T3 (P 0.05). Regarding the evaluation of goat semen samples, the motility parameters were consistent with the technique submitted; however, the IMP data did not appear as expected, requiring further evaluation for a better assessment of the technique for this species.Discussion: The data obtained from ram semen submitted to the immunosexing protocol, regarding the absolute evaluation of motility and IMP, demonstrated that the non-sexed semen (T1) was superior to the sexed treatments (T2 and T3); however, it is noteworthy that freezing started with approximately 50% of the cells, since the immunosexing technique results in a loss of viability of approximately 50% of the sperm, which corresponds to the ratio of sperm carrying the X chromosome. In addition, when the data in this study were transformed into relative values, no statistical differences were observed, indicating that the immunosexing protocol, as well as the freezing protocol, did not significantly affect the quality of ram sperm cells. In relation to the immunosexing of goat semen, future studies should be conducted in vitro to define a more appropriate protocol for the species and, in addition, in vivo studies should be performed to prove the quality of the technique. It was concluded that the immunosexing process using a monoclonal antibody against sex-specific protein (HY) associated with the use of powdered coconut water diluent (ACP101/102c) in the cryopreservation of semen proved to be efficient in the in vitro evaluation of ovine species

A carnosina diminui os efeitos da acidose muscular durante o exercício

A carnosina é um dipeptídeo composto pelos aminoácidos beta-alanina e histidina encontrado em maior parte no tecido muscular. Sua síntese ocorre através da enzima carnosina sintase tendo a beta alanina como precursor limitante. Estudos tem mostrado que o aumento nos níveis de carnosina melhora o desempenho esportivo, devido a redução da diminuição do pH intramuscular durante o exercício, proporcionando maior capacidade de tamponamento de íons de hidrogênio (H+). O objetivo da presente revisão foi analisar estudos relacionados aos efeitos da suplementação crônica de beta-alanina no desempenho esportivo nos últimos anos. A estratégia de busca foi realizada através de consulta às bases de dados PubMed e Trip database, utilizando os descritores “carnosineâ€, “beta-alanine†e “exercise†de forma combinada em artigos publicados entre janeiro de 2007 e dezembro de 2013. Foram incluídos na análise apenas artigos originais e foram excluídos artigos que avaliaram amostra não saudável, suplementação de beta-alanina associada a outras substâncias, além de teses, dissertações e trabalhos de revisão. Com base na reunião de informações acreditamos que a suplementação de beta alanina pode ser considerada uma estratégia eficaz no aumento dos níveis de carnosina, refletindo em melhoria de desempenho através de aumento do tempo de exaustão e atraso no limiar de fadiga neuromuscular, apresentando melhores efeitos em atividades de alta intensidade. ABSTRACTDoes carnosine reduce the effects of muscle acidosis during exercise?Carnosine is a dipeptide of beta-alanine and histidine amino acids, being found in most of the muscle tissue. Its synthesis occurs through carnosine synthase enzyme having the beta alanine as a limiting precursor. The most recent works have shown that increased levels of carnosine improves sports performance due to reducing of decrease in pH intramuscular during exercise, providing greater buffering capacity of hydrogen ions (H +). The aim of this review was to analyze works related to the effects of chronic beta-alanine supplementation on exercise performance recent years. The search strategy was done through consultation with PubMed and Trip Database data using the combined descriptors “carnosineâ€, “beta-alanine†and “exercise†in articles published between January 2007 and December 2013. We included in this analysis just original articles and articles that evaluated unhealthy sample, beta-alanine supplementation associated with other substances, as well as these, dissertations and reviews were excluded. Based on information meeting, we believe that beta-alanine supplementation can be considered an effective strategy in increasing carnosine levels, reflecting improved performance through increased time to exhaustion and delayed on neuromuscular fatigue threshold, presenting better effects in high intensity activities

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 methodology 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 using eosin-nigrosine dye, and the percentages of healthy and normal spermatozoa were determined. A bibliographic survey and a market study of similar products and technologies were carried out to provide an economic viability metanalysis of the bioproduct (ACP-102c) and bioprocess (immunosex). The data were analyzed using R-project©, and comparisons made between animals and between thawing periods using T test. There were no statistically significant differences between animals and between periods (P > 0.05), except for the normal sperm parameter, in which animal A1 had the lowest percentage (P < 0.05). As for the cost-benefit ratio, flow cytometry as a technique is more laborious and expensive, while immunosexing associated with cryopreservation in ACP-102c diluent has proven more practical, with regards to both sperm sexing techniques and diluents for sperm conservation.Discussion: In general, the quality of cryopreserved sexed semen was lower than that of non-sexed semen; however, in this study, both in the comparison between animals and between evaluation periods, similar values of motility, viability, and sperm morphology were obtained for sexed and several non-sexed cryopreserved semen samples, demonstrating that immunosexing did not severely affect the sperm structure, and that the ACP-102c conservation medium was efficient at maintaining the plasma membrane of these sperm. In the evaluation of economic aspects, it was observed that immunosexing, associated with cryopreservation in ACP-102c diluent, proved to be the most practical technique, requiring only conventional equipment, and allowing a greater field of application, since the immunosexing semen can be used for primiparous and multiparous females. Thus, it was concluded that immunosexing associated with cryopreservation in an ACP-102c diluent was more cost-effective, more practical, and had significantly improved sperm quality results after sexing

CRIOPRESERVAÇÃO DO SÊMEN OVINO EM MEIO DILUENTE À BASE DE ÁGUA DE COCO EM PÓ (ACP-102C)

The aim of this study was to evaluate the ACP-102c extender in the cryopreservation of ram semen compared to tris-glucose-egg yolk (TRIS) extender and fresh semen. Forty-eight ejaculates were collected from four rams and two aliquots per ejaculate were taken for dilution and cryopreservation in ACP-102c or TRIS and a third aliquot used for the fresh semen analysis. Either the fresh semen and cryopreserved in both extenders were evaluated for viability, integrity of plasma and acrosomal membrane, hypoosmotic swelling test, DNA fragmentation and sperm motility. The extenders did not differ for sperm viability, acrosome and plasma membrane integrity, DNA fragmentation and quantitative and qualitative parameters of sperm velocity after thawing, but differed in hypoosmotic swelling test, total and progressive motility and lateral extent of the head as well as in all motility parameters evaluated (except linearity and straightness) after two hours of incubation at 37 oC. There was variability among rams in total and progressive motility of semen cryopreserved in ACP-102c after thawing. The ACP-102c extender showed less protection in the cryopreservation of ram sperm when compared to TRIS, but the improvement in its formulation and freezing protocols may make it an alternative to freezing ram semen

Morphologic evaluation of goat spermatozoa diluted and frozen in media based on powder coconut water (PCW-101) or TRIS, stained by eosin-nigrosin and bromophenol blue AVALIAÇÃO MORFOLÓGICA DE ESPERMATOZÓIDES CAPRINOS DILUÍDOS E CONGELADOS EM MEIO À BASE DE ÁGUA DE COCO EM PÓ (ACP-101) OU TRIS, CORADOS POR EOSINA-NIGROSINA E AZUL DE BROMOFENOL

&lt;p&gt;This aims of the work were: to evaluate in vitro the goat semen frozen in diluents media based on powder coconut water (PCW-101) and TRIS and, compare the bromophenol blue stain efficiency with the eosin-nigrosin stain. The ejaculateds were divided and diluted into PCW-101 and TRIS, frozed and thawed after 30 days. Spermatic morphology was evaluated, through semen smears stained by eosin-nigrosin (EN) and bromophenol blue (BB). The morphologic parameters evaluated were: normal spermatozoa (N), head alteration (HA), intermediary piece alteration (IPA), tail alteration (TA), proximal citoplasmic drop (PCD), distal citoplasmic drop (DCD), and detached head (DH). There wasn’t significant difference in the observation of N between media, staining and their interactions after 5 minutes of thermo resistance test. After 120, the N was significantly influenced by media, where the TRIS presented better results. The incubation period of 120 minutes at 37ºC affect the spermatic morphology, increasing the HA percentages. The media based on TRIS promoted better protection from the cryoinjuries on frozen goat spermatozoa. BB staining was efficient on the fresh and post-thaw goat semen evaluation.&lt;br /&gt;&lt;br /&gt;KEY WORDS: Bromophenol blue, goat, powder coconut water; TRIS, spermatozoa.&lt;/p&gt; &lt;p&gt;Este trabalho teve como objetivos avaliar a morfologia de espermatozoides caprinos frescos e congelados em meios à base de água de coco em pó (ACP-101) e TRIS, bem como comparar a eficiência dos corantes azul de bromofenol (AB) e eosina-nigrosina (EN). Cada ejaculado foi dividido, diluído em ACP-101 e TRIS, congelado e, após trinta dias, descongelado. Analisou-se a morfologia espermática por esfregaços corados por EN e AB. Os parâmetros morfológicos foram: espermatozoides normais (N), alterações de cabeça (AC), de peça intermediária (API), de flagelo (AF), gotas citoplasmáticas proximal (GCP) e distal (GCD) e cabeça destacada (CD). Não se verificou diferença de N entre tratamentos após cinco minutos do teste de termorresistência. Após 120 minutos, a quantificação de N foi influenciada pelo diluente, tendo o TRIS resultados superiores. O período de incubação afetou a morfologia espermática, aumentando as porcentagens de AC. Conclui-se que o meio à base de TRIS promoveu maior proteção quanto às crioinjúrias em espermatozoides caprinos. O corante AB demonstrou ser mais eficiente na avaliação do sêmen caprino fresco e pós-descongelação.&lt;br /&gt;&lt;br /&gt;PALAVRAS-CHAVES: Água de coco em pó, azul de bromofenol, caprinos, espermatozoides, TRIS.&lt;/p&gt

A carnosina diminui os efeitos da acidose muscular durante o exercício

Does carnosine reduce the effects of muscle acidosis during exercise?Carnosine is a dipeptide of beta-alanine and histidine amino acids, being found in most of the muscle tissue. Its synthesis occurs through carnosine synthase enzyme having the beta alanine as a limiting precursor. The most recent works have shown that increased levels of carnosine improves sports performance due to reducing of decrease in pH intramuscular during exercise, providing greater buffering capacity of hydrogen ions (H +). The aim of this review was to analyze works related to the effects of chronic beta-alanine supplementation on exercise performance recent years. The search strategy was done through consultation with PubMed and Trip Database data using the combined descriptors “carnosine”, “beta-alanine” and “exercise” in articles published between January 2007 and December 2013. We included in this analysis just original articles and articles that evaluated unhealthy sample, beta-alanine supplementation associated with other substances, as well as these, dissertations and reviews were excluded. Based on information meeting, we believe that beta-alanine supplementation can be considered an effective strategy in increasing carnosine levels, reflecting improved performance through increased time to exhaustion and delayed on neuromuscular fatigue threshold, presenting better effects in high intensity activities.A carnosina é um dipeptídeo composto pelos aminoácidos beta-alanina e histidina encontrado em maior parte no tecido muscular. Sua síntese ocorre através da enzima carnosina sintase tendo a beta alanina como precursor limitante. Estudos tem mostrado que o aumento nos níveis de carnosina melhora o desempenho esportivo, devido a redução da diminuição do pH intramuscular durante o exercício, proporcionando maior capacidade de tamponamento de íons de hidrogênio (H+). O objetivo da presente revisão foi analisar estudos relacionados aos efeitos da suplementação crônica de beta-alanina no desempenho esportivo nos últimos anos. A estratégia de busca foi realizada através de consulta às bases de dados PubMed e Trip database, utilizando os descritores “carnosine”, “beta-alanine” e “exercise” de forma combinada em artigos publicados entre janeiro de 2007 e dezembro de 2013. Foram incluídos na análise apenas artigos originais e foram excluídos artigos que avaliaram amostra não saudável, suplementação de beta-alanina associada a outras substâncias, além de teses, dissertações e trabalhos de revisão. Com base na reunião de informações acreditamos que a suplementação de beta alanina pode ser considerada uma estratégia eficaz no aumento dos níveis de carnosina, refletindo em melhoria de desempenho através de aumento do tempo de exaustão e atraso no limiar de fadiga neuromuscular, apresentando melhores efeitos em atividades de alta intensidade.

AVALIAÇÃO DA CAPACIDADE REPRODUTIVA DE CAPRINOS TRATADOS COM TESTOSTERONA BIOIDÊNTICA POR VIA TRANSDÉRMICA

The objective of this study was to evaluate the effect of testosterone bioidentical by transdermal route of applicantion in goat scrotal, sexual behavior, quantitative and qualitative production of sperm, testosterone concentrations, FSH (Follicle Stimulating Hormone) and LH (luteinizing hormone) for the treatment of reproductive temporarily unable to play, since there is no data in the literature concerning association with hormone bioidêntico transdermal route of application in goats. We used four male goats (Saanen=03 and Anglo Nubian=01), with low libido and poor sperm quality, proven by andrological exam. Every week, over two months, it were taken and evalued sperm samples the scrotal circumference (SC) was mensure the bioidentical testosterone was applicated in the scrotal by transdermal route; blood samples were collected to determine serum testosterone, FSH and LH; beyond sexual behavior verification by testing libido. For purposes of data analysis used the statistical program SAS v.8 (2000). Serum testosterone showed significant differences depending on the treatment period, however, no significant difference was observed for FSH and LH between the different treatment periods. At the fresh and cooled sperm, the seminal parameters evaluated were not affected significantly throughout the experimental period. It can be concluded that the transdermal bioidentical testosterone application method in goats was effective, considering that it made the libido of treated animals more explicit, although despite the increase in serum testosterone levels, there was no interference in the seminal quality.O objetivo deste estudo foi avaliar o efeito da aplicação de testosterona bioidêntica, por via transdérmica escrotal em caprinos, sobre o comportamento sexual, a produção quanti-qualitativa dos espermatozoides e, as concentrações de testosterona, FSH (Hormônio Folículo Estimulante) e LH (Hormônio Luteinizante), para o tratamento de reprodutores temporariamente inaptos à reprodução, uma vez que não existem dados na literatura referentes à associação do hormônio bioidêntico com a via de aplicação transdérmica em caprinos. Foram utilizados quatro machos caprinos (Saanen=03 e Anglo Nubiano=01), apresentando baixa libido e qualidade seminal insatisfatória, comprovados por andrológico. Semanalmente, ao longo de dois meses, foram realizadas colheitas e avaliação do sêmen; mensuração da circunferência escrotal (CE); aplicação da testosterona bioidêntica por via transdérmica escrotal e colheita de sangue, para determinar concentração sérica de testosterona, FSH e LH; além da verificação do comportamento sexual através do teste de libido. Para efeito de análise dos dados utilizou-se o programa estatístico SAS v.8 (2000). As concentrações séricas de testosterona apresentaram diferença significativa em função do período de tratamento, entretanto, não foi observada diferença significativa para os valores de FSH e LH. Na análise, do sêmen fresco, e resfriado, os parâmetros seminais avaliados não foram influenciados significativamente ao longo do período experimental. Conclui-se que o método de aplicação da testosterona bioidêntica por via transdérmica em caprinos, foi eficaz, considerando-se que tornou mais explícita a exibição da libido dos animais tratados, embora apesar do aumento dos níveis séricos de testosterona, não houve nenhuma interferência na qualidade seminal

The Spectrophotometric method for determination of Colossoma macropomum (tambaqui) sperm concentration

To assist the reproductive management of tambaqui (Colossoma macropomum) males in laboratory and commercial fish farming, a linear regression model was obtained from concentration curves using the spectrophotometric method. Twenty-two tambaqui males with an average age of three years old were selected and divided into two groups containing 11 animals each. Both groups alternately received a single dose of carp pituitary extract (CPE; 2.0 mg/kg body weight, intracoelomic). Sperm was collected 14 h after hormonal treatment and diluted (1:4000; sperm:formaldehyde saline). The concentration was estimated by counting spermatozoa in a Neubauer chamber and by using a spectrophotometer (λ=540 nm). Individual sperm concentration ranged from 11.40 to 71.13 × 109 sperm/mL. The degree of transmittance ranged from 62.1% to 95.0%. There was a significant correlation (r2 = 0.966; p < 0.0001) between sperm concentration analyzed in a Neubauer chamber and transmittance at 540 nm. Analysis by spectrophotometry and the prediction provided by the equation Y=100.293 – 0.509X proved to be an efficient and fast method for estimating sperm concentration in tambaqui and can be used in routine procedures in artificial fish reproduction laboratories. Keywords: fish; reproduction; semen