RESUMO
Neste estudo, a avaliação do sêmen foi utilizada para investigar prováveis efeitos da presença do transgene sobre os parâmetros seminais de caprinos. Para tanto, amostras de sêmen foram colhidas semanalmente de três caprinos, sendo um não transgênico (NTG) e dois transgênicos (TG) para o Fator Estimulante de Colônia de Granulócitos humano (hG-CSF). Observou-se que, com exceção do volume ejaculado, não foram verificadas diferenças estatísticas para os dados relativos aos demais parâmetros: motilidade massal, concentração espermática, motilidade individual progressiva e patologias espermáticas. Concluiu-se que a presença do transgene hG-CSF não teve influência sobre os principais parâmetros seminais dos animais avaliados.
In this study, semen evaluation was used to investigate likely effects of transgene presence on seminal parameters of goats. For this purpose, semen samples were collected weekly from three goats, one non-transgenic (NTG) and two transgenic (TG) for the Human Granulocyte Colony Stimulating Factor (hG-CSF). It was observed that, with the exception of the ejaculated volume, no statistical differences were verified for the data related to the other parameters: mass motility, sperm concentration, individual progressive motility and spermatic pathologies. It was concluded that the presence of the hG-CSF transgene had no influence on the main seminal parameters of the animals evaluated.
Assuntos
Masculino , Animais , Animais Geneticamente Modificados , Fator Estimulador de Colônias de Granulócitos , Ruminantes/genética , Sêmen/fisiologiaRESUMO
Neste estudo, a avaliação do sêmen foi utilizada para investigar prováveis efeitos da presença do transgene sobre os parâmetros seminais de caprinos. Para tanto, amostras de sêmen foram colhidas semanalmente de três caprinos, sendo um não transgênico (NTG) e dois transgênicos (TG) para o Fator Estimulante de Colônia de Granulócitos humano (hG-CSF). Observou-se que, com exceção do volume ejaculado, não foram verificadas diferenças estatísticas para os dados relativos aos demais parâmetros: motilidade massal, concentração espermática, motilidade individual progressiva e patologias espermáticas. Concluiu-se que a presença do transgene hG-CSF não teve influência sobre os principais parâmetros seminais dos animais avaliados.(AU)
In this study, semen evaluation was used to investigate likely effects of transgene presence on seminal parameters of goats. For this purpose, semen samples were collected weekly from three goats, one non-transgenic (NTG) and two transgenic (TG) for the Human Granulocyte Colony Stimulating Factor (hG-CSF). It was observed that, with the exception of the ejaculated volume, no statistical differences were verified for the data related to the other parameters: mass motility, sperm concentration, individual progressive motility and spermatic pathologies. It was concluded that the presence of the hG-CSF transgene had no influence on the main seminal parameters of the animals evaluated.(AU)
Assuntos
Animais , Masculino , Ruminantes/genética , Animais Geneticamente Modificados , Fator Estimulador de Colônias de Granulócitos , Sêmen/fisiologiaRESUMO
Vários sistemas para a produção de proteínas recombinantes têm sido utilizados para a otimização daprodução das mesmas. A glândula mamária é considerada um sistema muito interessante para a produção destasproteínas devido ao seu alto nível de expressão e à sua capacidade para realizar modificações pós-traducionais.Caprinos podem produzir elevadas quantidades de leite durante um longo período de lactação e, portanto,tornam-se importantes candidatos como biorreatores para a expressão de proteínas recombinantes no leite. Noentanto, caprinos transgênicos não são de fácil obtenção devido à baixa eficiência dos métodos e à expressãoimprevisível da proteína recombinante. O incremento na eficiência dos métodos na transgênese em caprinos éum grande desafio a superar. Esta revisão tem por objetivo apresentar o estado atual dos métodos tradicionais,bem como apresentar técnicas emergentes para obtenção eficiente de caprinos transgênicos.(AU)
Aiming the optimization for recombinant protein production, various systems have been used. Themammary gland is considered to be a very interesting system for the production of these proteins due to its highlevel of expression and its ability to perform post-translational modifications. Goats can produce largequantities of milk over a long lactation period, and therefore this species is an important candidate forrecombinant protein expression in milk. However, it is not easy to generate transgenic goats due to the lowefficiency of methods and unpredictable expression of recombinant protein. An increase in efficiency fortransgenic methodologies for goats is a big challenge to overcome. This review aims to present the state of art ofthe traditional methods, as well as the emerging technologies for obtaining transgenic goats efficiently.(AU)
Assuntos
Animais , Feminino , Lactente , Ruminantes/genética , Ruminantes/metabolismo , Proteínas Recombinantes/análise , Glândulas Mamárias Animais , BiotecnologiaRESUMO
Vários sistemas para a produção de proteínas recombinantes têm sido utilizados para a otimização daprodução das mesmas. A glândula mamária é considerada um sistema muito interessante para a produção destasproteínas devido ao seu alto nível de expressão e à sua capacidade para realizar modificações pós-traducionais.Caprinos podem produzir elevadas quantidades de leite durante um longo período de lactação e, portanto,tornam-se importantes candidatos como biorreatores para a expressão de proteínas recombinantes no leite. Noentanto, caprinos transgênicos não são de fácil obtenção devido à baixa eficiência dos métodos e à expressãoimprevisível da proteína recombinante. O incremento na eficiência dos métodos na transgênese em caprinos éum grande desafio a superar. Esta revisão tem por objetivo apresentar o estado atual dos métodos tradicionais,bem como apresentar técnicas emergentes para obtenção eficiente de caprinos transgênicos.
Aiming the optimization for recombinant protein production, various systems have been used. Themammary gland is considered to be a very interesting system for the production of these proteins due to its highlevel of expression and its ability to perform post-translational modifications. Goats can produce largequantities of milk over a long lactation period, and therefore this species is an important candidate forrecombinant protein expression in milk. However, it is not easy to generate transgenic goats due to the lowefficiency of methods and unpredictable expression of recombinant protein. An increase in efficiency fortransgenic methodologies for goats is a big challenge to overcome. This review aims to present the state of art ofthe traditional methods, as well as the emerging technologies for obtaining transgenic goats efficiently.
Assuntos
Feminino , Animais , Lactente , Glândulas Mamárias Animais , Proteínas Recombinantes/análise , Ruminantes/genética , Ruminantes/metabolismo , BiotecnologiaRESUMO
In transgenic murine models, the study of certain organs or tissues can be performed after euthanasia of some specimens. However, this practice may not be economically feasible when applied to livestock such as transgenic goats. It is necessary to use minimally invasive methods to perform in vivo studies of organs that may be affected by disorders related to the activity of the transgene, particularly during milk production, when the recombinant protein is secreted. The aim of this study was to describe ultrasonographic findings of the liver, gallbladder, spleen, kidneys, and mammary glands in transgenic goats for evaluating the effect of human granulocyte-colony stimulating factor (hG-CSF) expression in milk during induced lactation. Six female Canindé goats-three transgenic (T) and three non-transgenic (NT)-were subjected to hormone therapy to induce lactation; ultrasonographic examinations of the liver, gallbladder, spleen, kidneys, and mammary gland were performed during both the hormonal therapy and the lactation period at different intervals depending on the organ being examined. On Day 16 (Day 1 = hormonal therapy initiation), all goats were lactating and presented healthy mammary glands, characterized by echogenic parenchyma showing a granular echotexture. Transgenic and nontransgenic goats were compared on the basis of measurements and ultrasound images obtained from each organ. No differences between T and NT animals were observed in the examined area for the liver, gallbladder, spleen, and kidneys. Liver and renal echogenicity and appearance of gallbladder and portal and hepatic veins were similar in all females. Ultrasonographic findings of the mammary gland, liver, gallbladder, spleen, and kidneys in transgenic goats did not show any difference from those in nontransgenic goats.[...](AU)
Em camundongos transgênicos, o estudo de certos órgãos ou tecidos pode ser viabilizado por meio da eutanásia de alguns exemplares. Entretanto, esta prática pode não ser economicamente viável quando aplicada a animais de produção, como caprinos transgênicos. É necessário utilizar métodos minimamente invasivos para estudar in vivo os órgãos que podem estar envolvidos em desordens relacionadas à atividade do transgene, principalmente durante a produção de leite, quando a proteína recombinante é secretada. O objetivo deste estudo foi descrever as características ultrassonográficas de fígado, vesícula biliar, baço, rins e glândula mamária de caprinos transgênicos expressando o fator estimulante de colônias de granulócitos humano (hG-CSF) no leite, durante lactação induzida. Seis fêmeas caprinas da raça Canindé: três não transgênicas (NT) e três transgênicas (T) foram submetidas a tratamento hormonal para indução da lactação; os exames ultrassonográficos de fígado, vesícula biliar, baço, rins e glândula mamária foram realizados durante tratamento hormonal e lactação, em diferentes intervalos de acordo com o órgão. No dia 16 (Dia 1 = início do tratamento hormonal) todas as cabras estavam em lactação e apresentando uma glândula mamária saudável, caracterizada por parênquima ecogênico, com uma ecotextura granular. As cabras T e NT foram comparadas em relação às mensurações e aparência ultrassonográfica obtida de cada órgão. Não foram observadas diferenças com relação à área para exame de fígado, vesícula biliar, baço e rins entre T e NT. A ecogenicidade de fígado e rins, a aparência da vesícula biliar e das veias porta e hepática foram similares em todas as fêmeas. As mensurações ultrassonográficas dos órgãos abdominais examinados não revelaram diferenças entre animais T e NT.[...](AU)
Assuntos
Animais , Ruminantes/anatomia & histologia , Hormônio do Crescimento Humano/efeitos adversos , Ultrassom/métodos , Lactação , Fígado , Vesícula Biliar , Baço , Rim , Glândulas Mamárias AnimaisRESUMO
In transgenic murine models, the study of certain organs or tissues can be performed after euthanasia of some specimens. However, this practice may not be economically feasible when applied to livestock such as transgenic goats. It is necessary to use minimally invasive methods to perform in vivo studies of organs that may be affected by disorders related to the activity of the transgene, particularly during milk production, when the recombinant protein is secreted. The aim of this study was to describe ultrasonographic findings of the liver, gallbladder, spleen, kidneys, and mammary glands in transgenic goats for evaluating the effect of human granulocyte-colony stimulating factor (hG-CSF) expression in milk during induced lactation. Six female Canindé goats-three transgenic (T) and three non-transgenic (NT)-were subjected to hormone therapy to induce lactation; ultrasonographic examinations of the liver, gallbladder, spleen, kidneys, and mammary gland were performed during both the hormonal therapy and the lactation period at different intervals depending on the organ being examined. On Day 16 (Day 1 = hormonal therapy initiation), all goats were lactating and presented healthy mammary glands, characterized by echogenic parenchyma showing a granular echotexture. Transgenic and nontransgenic goats were compared on the basis of measurements and ultrasound images obtained from each organ. No differences between T and NT animals were observed in the examined area for the liver, gallbladder, spleen, and kidneys. Liver and renal echogenicity and appearance of gallbladder and portal and hepatic veins were similar in all females. Ultrasonographic findings of the mammary gland, liver, gallbladder, spleen, and kidneys in transgenic goats did not show any difference from those in nontransgenic goats.[...]
Em camundongos transgênicos, o estudo de certos órgãos ou tecidos pode ser viabilizado por meio da eutanásia de alguns exemplares. Entretanto, esta prática pode não ser economicamente viável quando aplicada a animais de produção, como caprinos transgênicos. É necessário utilizar métodos minimamente invasivos para estudar in vivo os órgãos que podem estar envolvidos em desordens relacionadas à atividade do transgene, principalmente durante a produção de leite, quando a proteína recombinante é secretada. O objetivo deste estudo foi descrever as características ultrassonográficas de fígado, vesícula biliar, baço, rins e glândula mamária de caprinos transgênicos expressando o fator estimulante de colônias de granulócitos humano (hG-CSF) no leite, durante lactação induzida. Seis fêmeas caprinas da raça Canindé: três não transgênicas (NT) e três transgênicas (T) foram submetidas a tratamento hormonal para indução da lactação; os exames ultrassonográficos de fígado, vesícula biliar, baço, rins e glândula mamária foram realizados durante tratamento hormonal e lactação, em diferentes intervalos de acordo com o órgão. No dia 16 (Dia 1 = início do tratamento hormonal) todas as cabras estavam em lactação e apresentando uma glândula mamária saudável, caracterizada por parênquima ecogênico, com uma ecotextura granular. As cabras T e NT foram comparadas em relação às mensurações e aparência ultrassonográfica obtida de cada órgão. Não foram observadas diferenças com relação à área para exame de fígado, vesícula biliar, baço e rins entre T e NT. A ecogenicidade de fígado e rins, a aparência da vesícula biliar e das veias porta e hepática foram similares em todas as fêmeas. As mensurações ultrassonográficas dos órgãos abdominais examinados não revelaram diferenças entre animais T e NT.[...]
Assuntos
Animais , Baço , Fígado , Glândulas Mamárias Animais , Hormônio do Crescimento Humano/efeitos adversos , Lactação , Rim , Ruminantes/anatomia & histologia , Ultrassom/métodos , Vesícula BiliarRESUMO
The aim of this study was to report the occurrence of pregnancy loss in transgenic goat submitted toprenatal ultrasonography monitoring. We used on goat transgenic for human Granulocyte Colony StimulatingFactor (hG-CSF). The animal was submitted to mating after estrous synchronization. Ultrasound examinationswere carried out until D60 of pregnancy by transrectal via and after D60 by transabdominal via. Someparameters were observed such as morphology, organogenesis and formation of skeletal fetuses, viability withcardiac activity and fetus movements. During all sonographic evaluations, the conceptus showed normaldevelopment of their morphology and viability. Goat had fetal development completed by D150, however, therewere problems at birth and the fetus has died. In conclusion, the transgenic fetus remained viable and showednormal development during pregnancy, although still birth.(AU)
Assuntos
Animais , Feminino , Cabras/embriologia , Gravidez , Ultrassom/métodos , Fator Estimulador de Colônias de Granulócitos/administração & dosagemRESUMO
The aim of this study was to report the occurrence of pregnancy loss in transgenic goat submitted toprenatal ultrasonography monitoring. We used on goat transgenic for human Granulocyte Colony StimulatingFactor (hG-CSF). The animal was submitted to mating after estrous synchronization. Ultrasound examinationswere carried out until D60 of pregnancy by transrectal via and after D60 by transabdominal via. Someparameters were observed such as morphology, organogenesis and formation of skeletal fetuses, viability withcardiac activity and fetus movements. During all sonographic evaluations, the conceptus showed normaldevelopment of their morphology and viability. Goat had fetal development completed by D150, however, therewere problems at birth and the fetus has died. In conclusion, the transgenic fetus remained viable and showednormal development during pregnancy, although still birth.
Assuntos
Feminino , Animais , Cabras/embriologia , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Gravidez , Ultrassom/métodosRESUMO
Background: The production of transgenic animals has been envisioned as a viable strategy to improve food quality, animal yield, and for the production of bioproducts that can be used for the benefit of the human and animal population. Transgenic animals have been used to improve production traits, to add value to animal products, to minimize the impact on the environment, to promote disease resistance, and most notably, to produce recombinant proteins in natural fluids, such as milk, that can be collected, purified and used as biomedical products (biopharming). This review aims to discuss past and recent technological advances in animal transgenesis, and the perspective for biopharming in Brazil.Review: Since the production of recombinant human insulin from Escherichia coli in the 1970s, continuous development of new platforms has allowed a significant expansion in the biopharmaceutical market. The animal platform has been shown to be highly competitive by adding value as low cost implementation, production and scale up, as well as high productivity of synthesized proteins. The expression of recombinant proteins in milk represents the most developed system for production of biopharmaceutical drugs in animals, with two approved biopharmaceuticals for human use: Atryn®, a recombinant antithrombin produced in the milk of goats, approved in 2006 by European Medicines Agency (EMA) and in 2009 by US Food and Drug Administration (FDA), and more recently, Ruconest®, a recombinant human C1 esterase inhibitor protein (C1INH) produced in the milk of rabbits, first approved by EMA in 2012, followed by the FDA approval in 2014. Transgenic animals have been produced by many strategies that have gradually evolved over the decades, including the use of embryo microinjection, viral vectors and transposable elements, sperm-mediated gene transfer, and cloning by somatic cell nuclear transfer (SCNT).[...]
Assuntos
Animais , Animais Geneticamente Modificados , Produtos Biológicos , Proteínas Recombinantes/uso terapêutico , Brasil , Clonagem de Organismos , Glândulas Mamárias Animais , Microinjeções/veterináriaRESUMO
Background: The production of transgenic animals has been envisioned as a viable strategy to improve food quality, animal yield, and for the production of bioproducts that can be used for the benefit of the human and animal population. Transgenic animals have been used to improve production traits, to add value to animal products, to minimize the impact on the environment, to promote disease resistance, and most notably, to produce recombinant proteins in natural fluids, such as milk, that can be collected, purified and used as biomedical products (biopharming). This review aims to discuss past and recent technological advances in animal transgenesis, and the perspective for biopharming in Brazil.Review: Since the production of recombinant human insulin from Escherichia coli in the 1970s, continuous development of new platforms has allowed a significant expansion in the biopharmaceutical market. The animal platform has been shown to be highly competitive by adding value as low cost implementation, production and scale up, as well as high productivity of synthesized proteins. The expression of recombinant proteins in milk represents the most developed system for production of biopharmaceutical drugs in animals, with two approved biopharmaceuticals for human use: Atryn®, a recombinant antithrombin produced in the milk of goats, approved in 2006 by European Medicines Agency (EMA) and in 2009 by US Food and Drug Administration (FDA), and more recently, Ruconest®, a recombinant human C1 esterase inhibitor protein (C1INH) produced in the milk of rabbits, first approved by EMA in 2012, followed by the FDA approval in 2014. Transgenic animals have been produced by many strategies that have gradually evolved over the decades, including the use of embryo microinjection, viral vectors and transposable elements, sperm-mediated gene transfer, and cloning by somatic cell nuclear transfer (SCNT).[...](AU)
Assuntos
Animais , Animais Geneticamente Modificados , Proteínas Recombinantes/uso terapêutico , Produtos Biológicos , Brasil , Microinjeções/veterinária , Clonagem de Organismos , Glândulas Mamárias AnimaisRESUMO
The objective of this study was to evaluate the development of transgenic (T) goat embryos and fetuses for human Granulocyte Colony Stimulating Factor (hG-CSF) by ultrasonography. Four pregnancies in non-transgenic (NT) goats were obtained after fertilization (either fixed-time artificial insemination or natural mating) using the T male for hG-CSF. Ultrasound examinations were carried out at 30, 40 (transrectal via), 50, 60, 90 and 120 days of pregnancy (transabdominal via). Some parameters were observed such as morphology, organogenesis and formation of skeletal fetuses, viability with cardiac activity and fetuses movements. Measurements were taken of the crown-rump length, diameter of embryonic vesicle, thorax, abdomen, umbilical cord and placentomes. After parturition, DNA testing was conducted in all offspring and 4 T and 2 NT kids were identified. The conceptus started their differentiation at 40 days. The heart was detected in all examinations and the heart chambers were assessed at 50 days. Gastric compartments, liver and kidneys were observed at 60 days, the same period that all bony structures were visualized. Average values of all evaluated parameters had a gradual increase with the progression of pregnancy. T and NT goat embryos and fetuses had a similar growth and all remained viable throughout the experimental period.(AU)
O objetivo deste estudo foi avaliar o desenvolvimento de embriões e fetos transgênicos (T) para o Fator Estimulante de Colôniasde Granulócitos humano (hG-CSF) por ultrassonografia. Quatro gestações em cabras não transgênicas (NT) foram obtidas pos fecundação (inseminação artificial em tempo fixo ou monta controlada) utilizando o bode T para o hG-CSF. Exames ultrassonográficos foram realizados aos 30, 40 (via transretal), 50, 60, 90 e 120 dias de gestação (via transabdominal). Alguns parâmetros foram observados como morfologia, organogênese e formação do esqueleto fetal, viabilidade por meio de atividade cardíaca e movimento fetal. As seguintes mensurações foram realizadas: comprimento crânio caudal, diâmetro da vesícula embrionária, do tórax, do abdomen, do cordão umbilical e dos placentomas. Após o parto, o exame por PCR foi conduzido em todas as crias e 4 T e 2NT foram identificadas. O concepto iniciou sua diferenciação aos 40 dias. O coração foi detectado em todos os exames e as câmeras cardíacas foram identificadas aos 50 dias. Compartimentos gástricos, fígado e rins foram observados aos 60 dias, o mesmo período que todas as estruturas ósseas foram visualizadas. Valores médios de todos os parâmetros avaliados tiveram um aumento gradual com o avanço da gestação. Embriões e fetos T e NT tiveram um crescimento similar e todos permaneceram viáveis durante o período experimental.(AU)
Assuntos
Animais , Feminino , Cabras/embriologia , Cabras/anatomia & histologia , Ultrassonografia/veterinária , Organismos Geneticamente Modificados , Fator Estimulador de Colônias de Granulócitos , Reação em Cadeia da Polimerase/veterinária , Feto/diagnóstico por imagemRESUMO
The objective of this study was to evaluate the development of transgenic (T) goat embryos and fetuses for human Granulocyte Colony Stimulating Factor (hG-CSF) by ultrasonography. Four pregnancies in non-transgenic (NT) goats were obtained after fertilization (either fixed-time artificial insemination or natural mating) using the T male for hG-CSF. Ultrasound examinations were carried out at 30, 40 (transrectal via), 50, 60, 90 and 120 days of pregnancy (transabdominal via). Some parameters were observed such as morphology, organogenesis and formation of skeletal fetuses, viability with cardiac activity and fetuses movements. Measurements were taken of the crown-rump length, diameter of embryonic vesicle, thorax, abdomen, umbilical cord and placentomes. After parturition, DNA testing was conducted in all offspring and 4 T and 2 NT kids were identified. The conceptus started their differentiation at 40 days. The heart was detected in all examinations and the heart chambers were assessed at 50 days. Gastric compartments, liver and kidneys were observed at 60 days, the same period that all bony structures were visualized. Average values of all evaluated parameters had a gradual increase with the progression of pregnancy. T and NT goat embryos and fetuses had a similar growth and all remained viable throughout the experimental period.
O objetivo deste estudo foi avaliar o desenvolvimento de embriões e fetos transgênicos (T) para o Fator Estimulante de Colôniasde Granulócitos humano (hG-CSF) por ultrassonografia. Quatro gestações em cabras não transgênicas (NT) foram obtidas pos fecundação (inseminação artificial em tempo fixo ou monta controlada) utilizando o bode T para o hG-CSF. Exames ultrassonográficos foram realizados aos 30, 40 (via transretal), 50, 60, 90 e 120 dias de gestação (via transabdominal). Alguns parâmetros foram observados como morfologia, organogênese e formação do esqueleto fetal, viabilidade por meio de atividade cardíaca e movimento fetal. As seguintes mensurações foram realizadas: comprimento crânio caudal, diâmetro da vesícula embrionária, do tórax, do abdomen, do cordão umbilical e dos placentomas. Após o parto, o exame por PCR foi conduzido em todas as crias e 4 T e 2NT foram identificadas. O concepto iniciou sua diferenciação aos 40 dias. O coração foi detectado em todos os exames e as câmeras cardíacas foram identificadas aos 50 dias. Compartimentos gástricos, fígado e rins foram observados aos 60 dias, o mesmo período que todas as estruturas ósseas foram visualizadas. Valores médios de todos os parâmetros avaliados tiveram um aumento gradual com o avanço da gestação. Embriões e fetos T e NT tiveram um crescimento similar e todos permaneceram viáveis durante o período experimental.
Assuntos
Feminino , Animais , Cabras/anatomia & histologia , Cabras/embriologia , Fator Estimulador de Colônias de Granulócitos , Organismos Geneticamente Modificados , Ultrassonografia/veterinária , Feto/diagnóstico por imagem , Reação em Cadeia da Polimerase/veterináriaRESUMO
Milk production of transgenic does was evaluated by ultrasound measurements of the mammary gland. Two Canindé goats, which were nine months of age were used in the trial, one non-transgenic or other transgenic for hG-CSF. For hormone-induced lactation, animals were given estradiol (0.25mg/kg, IM), progesterone (0.75mg/kg, IM), and prednisolone (0.4mg/kg, IM). Ultrasonographic exams were carried out during milking, using a Falcon 100 ultrasound equipment with a 5MHz convex probe and were performed by the same operator. The results were expressed as mean±standard error. The maximum greater length and shorter length of the cistern were respectively 5.14cm and 1.36cm for the transgenic animal and 7.28cm and 2.25cm for non-transgenic, which is consistent with the maximum milk volume produced. The relationship between the average area of cisterns and milk yield was expressed as a linear correlation curve, with a correlation coefficient significantly positive for both transgenic (Y=-1.1314+10.8538*x; r=0.97) and non-transgenic (Y=-21.7551+18.3634*x; r=0.97) animals. In conclusion, the ultrasound is a practice and appropriate technique to evaluate the cisterns in ruminant udders in transgenic animal.
RESUMO
Milk production of transgenic does was evaluated by ultrasound measurements of the mammary gland. Two Canindé goats, which were nine months of age were used in the trial, one non-transgenic or other transgenic for hG-CSF. For hormone-induced lactation, animals were given estradiol (0.25mg/kg, IM), progesterone (0.75mg/kg, IM), and prednisolone (0.4mg/kg, IM). Ultrasonographic exams were carried out during milking, using a Falcon 100 ultrasound equipment with a 5MHz convex probe and were performed by the same operator. The results were expressed as mean±standard error. The maximum greater length and shorter length of the cistern were respectively 5.14cm and 1.36cm for the transgenic animal and 7.28cm and 2.25cm for non-transgenic, which is consistent with the maximum milk volume produced. The relationship between the average area of cisterns and milk yield was expressed as a linear correlation curve, with a correlation coefficient significantly positive for both transgenic (Y=-1.1314+10.8538*x; r=0.97) and non-transgenic (Y=-21.7551+18.3634*x; r=0.97) animals. In conclusion, the ultrasound is a practice and appropriate technique to evaluate the cisterns in ruminant udders in transgenic animal.(AU)
Assuntos
Animais , Ovinos/crescimento & desenvolvimento , Animais Geneticamente Modificados , Glândulas Mamárias Animais , Ultrassonografia/veterinária , Biometria/métodosRESUMO
In order to produce transgenic goats with hG-CSF, a total of 24 adult Saanen and 48 adult undefined breed goats were used as donors and recipients, respectively. Donors were estrus-synchronized with vaginal sponges and superovulated by a treatment with 200 mg FSH given twice daily in decreasing doses over 3 days starting 48 h before sponge removal. Ovulation was induced by injecting 100µg GnRH 36 h after sponge removal. The recipients also received an estrus synchronization treatment. Donors were mated with fertile Saanen bucks and, approximately 72 h after sponge removal, zygotes were recovered surgically by flushing oviducts. The recovered zygotes were briefly centrifuged to a reliable visualization of the pronuclei. The DNA construct containing hG-CSF gene flanked by goat and bovine alphas1-casein sequences was injected into pronuclei of 129 zygotes. The microinjected embryos (3-6 per female) were transferred to 27 recipients. Ten recipients became pregnant and 12 kids were born. One transgenic male founder was identified in the group of kids. This is the first report of a birth of a transgenic goat in Latin America.
A fim de produzir caprinos transgênicos para o hG-CSF, utilizou-se 24 cabras Saanen adultas e 48 cabras sem raça definida adultas como doadoras e receptoras, respectivamente. As doadoras tiveram o estro sincronizado por esponjas vaginais e foram superovuladas com 200 mg de FSH em doses decrescentes, duas vezes ao dia e iniciando 48 h antes da retirada da esponja. A ovulação foi induzida pela injeção de 100 µg de GnRH às 36 h após a retirada da esponja. As receptoras também receberam um tratamento de sincronização do estro. As doadoras foram cobertas por bodes Saanen férteis e, aproximadamente 72 h após a retirada da esponja, os zigotos foram colhidos cirurgicamente por lavagem dos ovidutos. Os zigotos colhidos foram rapidamente centrifugados para uma melhor visualização dos pró-núcleos. A construção de DNA, contendo o gene do hG-CSF flanqueado pelos genes caprino e bovino da alfas1-caseína, foi injetada em 129 embriões. Os embriões microinjetados (3 a 6 por receptora) foram transferidos para 27 receptoras que responderam ao tratamento. Dez receptoras ficaram gestantes e 12 crias foram produzidas. Um macho transgênico fundador foi identificado no grupo de crias nascidas. Este é o primeiro relato do nascimento de um caprino transgênico na América Latina.