Neoplasms in domestic hamsters in Southern Brazil: epidemiological and pathological aspects of 40 cases1 / Neoplasias em hamsters domésticos no Sul do Brasil: aspectos epidemiológicos e patológicos de 40 casos

Although neoplasms are commonly reported in domestic hamsters, retrospective studies approaching spontaneous tumors with data regarding epidemiological findings are scarce. The present study aimed to describe the epidemiological and pathological findings of 40 cases of tumors in domestic hamsters diagnosed in a veterinary pathology laboratory in Southern Brazil from 2002 to 2019. Chinese hamster (Cricetulus griseus) was the most commonly affected species (16/40), followed by Djungarian hamster (Phodopus sungorus, 11/40) and Syrian hamster (Mesocricetus auratus, 4/40). Among the cases, 57.5% were females (23/40), while 42.5% were males (17/40). The affected hamsters' median age was of 14-months old, with an age range of 8- to 36-months old. Twenty-four cases were assessed as anatomopathological samples (biopsies), while 16 were composed of carcasses submitted to postmortem examination, and, therefore, the neoplasm was related to the cause of death. The integumentary system was frequently affected (60%, 24/40), followed by the female reproductive tract (22.5%, 9/40), hematopoietic system (10%, 4/40), digestive tract (5%, 2/40), and endocrine system (2.5%, 1/40). The most frequent neoplasm was squamous cell carcinoma (35%, 14/40), mostly on the lip/nasal region (50%, 7/14). Other tumors included fibrosarcoma (10%, 4/40), lymphoma (10%, 4/40), mammary cystadenoma (10%, 4/40), apocrine sweat gland adenoma (7.5%, 3/40), hemangiosarcoma (5%, 2/40), leiomyosarcoma (5%, 2/40), and granulosa ovarian cell tumor (5%, 2/40). The five remaining cases occurred individually and were composed of hepatoid gland adenoma, solid thyroid carcinoma, cutaneous melanoma, ovarian teratoma, and cutaneous trichoblastoma. Neoplasms were identified as an important cause of death and major reason to perform biopsy in domestic hamsters in Southern Brazil.(AU)

Embora neoplasmas em hamsters domésticos sejam comumente relatados, estudos retrospectivos abordando neoplasias espontâneas e os dados epidemiológicos associados são escassos. O presente estudo teve o objetivo de descrever os principais achados epidemiológicos e patológicos de 40 casos de tumores em hamsters domésticos diagnosticados em um laboratório de patologia veterinária do Sul do Brasil de 2002 a 2019. A principal espécie acometida foi o hamster chinês (Cricetulus griseus, 16/40), seguido por hamster anão russo siberiano (Phodopus sungorus, 11/40) e hamster sírio (Mesocricetus auratus, 4/40). As fêmeas corresponderam a 57,5% dos casos (23/40), enquanto os machos representaram 42,5% (17/40). Foram afetados roedores com uma faixa etária de 8 a 36 meses de idade, e uma mediana de 14 meses. Do total de casos, 24 foram exames anatomopatológicos (biopsias) e 16 casos foram examinados através de necropsia e, portanto, relacionados com a causa da morte dos animais. O sistema tegumentar foi o mais frequentemente acometido (60%; 24/40), seguido pelo trato reprodutivo (22,5%; 9/40), sistema hematopoietico (10%; 4/40), trato digestório (5%; 2/40) e sistema endócrino (2,5%; 1/40). A neoplasia mais frequentemente diagnosticada foi o carcinoma de células escamosas (35%; 14/40), localizado principalmente em região labionasal (50%; 7/14). Outros tumores incluíram fibrossarcoma (10%; 4/40), linfoma (10%; 4/40), adenoma cístico de glândula mamária (10%; 4/40), adenoma de glândula sudorípara (7,5%; 3/40), hemangiossarcoma (5%; 2/40), leiomiossarcoma (5%; 2/40) e tumor de células da granulosa (5%; 2/40). Os outros cinco casos remanescentes ocorreram individualmente e eram compostos por adenoma de glândula hepatoide, carcinoma sólido de tireoide, melanoma cutâneo, teratoma ovariano e tricoblastoma cutâneo. Neoplasmas foram identificados como importantes causas de morte ou razões para realização de biopsia em hamsters domésticos no Sul do Brasil.(AU)

Neoplasms in domestic hamsters in Southern Brazil: epidemiological and pathological aspects of 40 cases1 / Neoplasias em hamsters domésticos no Sul do Brasil: aspectos epidemiológicos e patológicos de 40 casos

Although neoplasms are commonly reported in domestic hamsters, retrospective studies approaching spontaneous tumors with data regarding epidemiological findings are scarce. The present study aimed to describe the epidemiological and pathological findings of 40 cases of tumors in domestic hamsters diagnosed in a veterinary pathology laboratory in Southern Brazil from 2002 to 2019. Chinese hamster (Cricetulus griseus) was the most commonly affected species (16/40), followed by Djungarian hamster (Phodopus sungorus, 11/40) and Syrian hamster (Mesocricetus auratus, 4/40). Among the cases, 57.5% were females (23/40), while 42.5% were males (17/40). The affected hamsters' median age was of 14-months old, with an age range of 8- to 36-months old. Twenty-four cases were assessed as anatomopathological samples (biopsies), while 16 were composed of carcasses submitted to postmortem examination, and, therefore, the neoplasm was related to the cause of death. The integumentary system was frequently affected (60%, 24/40), followed by the female reproductive tract (22.5%, 9/40), hematopoietic system (10%, 4/40), digestive tract (5%, 2/40), and endocrine system (2.5%, 1/40). The most frequent neoplasm was squamous cell carcinoma (35%, 14/40), mostly on the lip/nasal region (50%, 7/14). Other tumors included fibrosarcoma (10%, 4/40), lymphoma (10%, 4/40), mammary cystadenoma (10%, 4/40), apocrine sweat gland adenoma (7.5%, 3/40), hemangiosarcoma (5%, 2/40), leiomyosarcoma (5%, 2/40), and granulosa ovarian cell tumor (5%, 2/40). The five remaining cases occurred individually and were composed of hepatoid gland adenoma, solid thyroid carcinoma, cutaneous melanoma, ovarian teratoma, and cutaneous trichoblastoma. Neoplasms were identified as an important cause of death and major reason to perform biopsy in domestic hamsters in Southern Brazil.(AU)

Embora neoplasmas em hamsters domésticos sejam comumente relatados, estudos retrospectivos abordando neoplasias espontâneas e os dados epidemiológicos associados são escassos. O presente estudo teve o objetivo de descrever os principais achados epidemiológicos e patológicos de 40 casos de tumores em hamsters domésticos diagnosticados em um laboratório de patologia veterinária do Sul do Brasil de 2002 a 2019. A principal espécie acometida foi o hamster chinês (Cricetulus griseus, 16/40), seguido por hamster anão russo siberiano (Phodopus sungorus, 11/40) e hamster sírio (Mesocricetus auratus, 4/40). As fêmeas corresponderam a 57,5% dos casos (23/40), enquanto os machos representaram 42,5% (17/40). Foram afetados roedores com uma faixa etária de 8 a 36 meses de idade, e uma mediana de 14 meses. Do total de casos, 24 foram exames anatomopatológicos (biopsias) e 16 casos foram examinados através de necropsia e, portanto, relacionados com a causa da morte dos animais. O sistema tegumentar foi o mais frequentemente acometido (60%; 24/40), seguido pelo trato reprodutivo (22,5%; 9/40), sistema hematopoietico (10%; 4/40), trato digestório (5%; 2/40) e sistema endócrino (2,5%; 1/40). A neoplasia mais frequentemente diagnosticada foi o carcinoma de células escamosas (35%; 14/40), localizado principalmente em região labionasal (50%; 7/14). Outros tumores incluíram fibrossarcoma (10%; 4/40), linfoma (10%; 4/40), adenoma cístico de glândula mamária (10%; 4/40), adenoma de glândula sudorípara (7,5%; 3/40), hemangiossarcoma (5%; 2/40), leiomiossarcoma (5%; 2/40) e tumor de células da granulosa (5%; 2/40). Os outros cinco casos remanescentes ocorreram individualmente e eram compostos por adenoma de glândula hepatoide, carcinoma sólido de tireoide, melanoma cutâneo, teratoma ovariano e tricoblastoma cutâneo. Neoplasmas foram identificados como importantes causas de morte ou razões para realização de biopsia em hamsters domésticos no Sul do Brasil.(AU)

Isolation and characterization of the human ovarian cell population for transplantation into an artificial ovary

To support survival and growth of follicles, the transplantable artificial ovary should mimic the original organ, offering a physical (3D matrix) and biological support (cells). In order to replicate the ovarian cell populations, the aim of this study is to assess the proportions of stromal and endothelial cells in the ovarian cortex. To this end, ovarian biopsies were obtained from six women (mean age: 49 years). The epithelial layer and medulla were carefully removed. The cortex was finely minced and enzymatically digested and the isolated cells were fixed. For cell characterization, immunostaining for CD31 (for endothelial cells) and inhibin-α (for granulosa cells) was performed. Positive cells in each staining were counted and the proportion of the different cell populations was estimated from the total number of isolated cells. Since there is no specific marker for ovarian stromal cells, we estimated the proportion of these cells by performing a vimentin immunostaining and subtracting the proportions of CD31- and inhibin-α-positive cells. Immunostaining showed that 84% of isolated cells were vimentin-positive. From this pool, 3% were endothelial cells and 1% granulosa cells. Consequently, the population of ovarian stromal cells was 80%. In conclusion, our findings show that stromal cells represent the larger population of cells in the human ovarian cortex. While this ensures follicle survival and development in a normal ovary, we believe that the low proportion of endothelial cells could have a negative impact on the angiogenesis in the artificial ovary after the first days of transplantation.

Isolation and characterization of the human ovarian cell population for transplantation into an artificial ovary

To support survival and growth of follicles, the transplantable artificial ovary should mimic the original organ, offering a physical (3D matrix) and biological support (cells). In order to replicate the ovarian cell populations, the aim of this study is to assess the proportions of stromal and endothelial cells in the ovarian cortex. To this end, ovarian biopsies were obtained from six women (mean age: 49 years). The epithelial layer and medulla were carefully removed. The cortex was finely minced and enzymatically digested and the isolated cells were fixed. For cell characterization, immunostaining for CD31 (for endothelial cells) and inhibin-α (for granulosa cells) was performed. Positive cells in each staining were counted and the proportion of the different cell populations was estimated from the total number of isolated cells. Since there is no specific marker for ovarian stromal cells, we estimated the proportion of these cells by performing a vimentin immunostaining and subtracting the proportions of CD31- and inhibin-α-positive cells. Immunostaining showed that 84% of isolated cells were vimentin-positive. From this pool, 3% were endothelial cells and 1% granulosa cells. Consequently, the population of ovarian stromal cells was 80%. In conclusion, our findings show that stromal cells represent the larger population of cells in the human ovarian cortex. While this ensures follicle survival and development in a normal ovary, we believe that the low proportion of endothelial cells could have a negative impact on the angiogenesis in the artificial ovary after the first days of transplantation.(AU)

Vitrification of Ovarian Tissue from Non-Human Primates

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim

Vitrification of ovarian tissue from non-human primates

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened primate species and the needs to preserve their habitat, but also their gametes, development of preservation protocols are needed. Among the procedures, vitrification appears as a practical method to be applied in the near future. Although a low number of studies is reported, most of them were performed in the recent years. In this context, this article reviews recent information on the vitrification of ovarian tissue of non-human primates. Due to the limited number of studies in these species, observed data are compared with the literature in domestic and human mammals.[...]

Vitrification of ovarian tissue from non-human primates

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened primate species and the needs to preserve their habitat, but also their gametes, development of preservation protocols are needed. Among the procedures, vitrification appears as a practical method to be applied in the near future. Although a low number of studies is reported, most of them were performed in the recent years. In this context, this article reviews recent information on the vitrification of ovarian tissue of non-human primates. Due to the limited number of studies in these species, observed data are compared with the literature in domestic and human mammals.[...](AU)

Selection of pacu females to hormonal induction: effect of age and of evaluation methods / Seleção de fêmeas de pacu para indução hormonal: efeito da idade e dos métodos de avaliação

In this study, we investigated the reproductive cycle and spawning season of old (OFs) and young (YFs) P. mesopotamicus females, aiming to improve the process of selection of suitable fish for induced spawning. We also evaluated the accuracy of using external body characteristics in selecting suitable females. To that, 60 OFs and 60 YFs (10 and four years old, respectively) were submitted to seven samplings during two reproductive cycles. In each sampling, females (five to 10 per treatment) were randomly chosen to evaluation of plasma concentrations of gonadal steroids and ovary composition (biopsies and stereology). Females were also submitted to an external evaluation concerning the length of abdominal thickness, the degree of the abdominal rigidity and the aspect of urogenital papilla. In spite of OFs have shown reduced concentrations of 17α- hydroxyprogesterone during the spawning season, we observed that OFs and YFs kept in captivity had similar reproductive cycles and spawning seasons (in December for both groups). We do not recommend the exclusive use of external evaluation for selecting breeders to spawning induction due to a great inaccuracy of data and lack of association with histology and ovarian biopsy, which are more accurate and reliable.(AU)

Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêmeas “jovens” (FJs) de P. mesopotamicus, visando melhorar o processo de seleção de peixes para a indução hormonal. Avaliamos também a acurácia do uso de características corporais externas para a seleção de fêmeas para indução hormonal. Para isso, 60 FVs e 60 FJs (10 e quatro anos, respectivamente) foram submetidas a sete amostragens durante dois ciclos reprodutivos. Em cada amostra, as fêmeas (cinco a 10 por tratamento) foram capturadas para avaliação das concentrações plasmáticas de esteroides gonadais e composição dos ovários (biópsias e estereologia). As fêmeas foram também submetidas à uma avaliação externa relativa à espessura abdominal, o grau de rigidez abdominal e o aspecto da papila urogenital. Exceto pela ocorrência de concentrações reduzidas de 17α-hidroxiprogesterona nas FVs, durante a época de desova, observamos que FVs e FJs, mantidas em cativeiro, apresentam ciclos reprodutivos e épocas de desova similares (em dezembro para ambos os grupos). Não recomendamos o uso exclusivo da avaliação externa para a seleção de reprodutores para indução a desova, devido a uma imprecisão dos dados e falta de associação destes com a histologia e biópsia de ovário, os quais são mais precisos e confiáveis.(AU)

Selection of pacu females to hormonal induction: effect of age and of evaluation methods / Seleção de fêmeas de pacu para indução hormonal: efeito da idade e dos métodos de avaliação

In this study, we investigated the reproductive cycle and spawning season of old (OFs) and young (YFs) P. mesopotamicus females, aiming to improve the process of selection of suitable fish for induced spawning. We also evaluated the accuracy of using external body characteristics in selecting suitable females. To that, 60 OFs and 60 YFs (10 and four years old, respectively) were submitted to seven samplings during two reproductive cycles. In each sampling, females (five to 10 per treatment) were randomly chosen to evaluation of plasma concentrations of gonadal steroids and ovary composition (biopsies and stereology). Females were also submitted to an external evaluation concerning the length of abdominal thickness, the degree of the abdominal rigidity and the aspect of urogenital papilla. In spite of OFs have shown reduced concentrations of 17α- hydroxyprogesterone during the spawning season, we observed that OFs and YFs kept in captivity had similar reproductive cycles and spawning seasons (in December for both groups). We do not recommend the exclusive use of external evaluation for selecting breeders to spawning induction due to a great inaccuracy of data and lack of association with histology and ovarian biopsy, which are more accurate and reliable.

Neste estudo, investigamos o ciclo reprodutivo e a época de desova de fêmeas “velhas” (FVs) e fêmeas “jovens” (FJs) de P. mesopotamicus, visando melhorar o processo de seleção de peixes para a indução hormonal. Avaliamos também a acurácia do uso de características corporais externas para a seleção de fêmeas para indução hormonal. Para isso, 60 FVs e 60 FJs (10 e quatro anos, respectivamente) foram submetidas a sete amostragens durante dois ciclos reprodutivos. Em cada amostra, as fêmeas (cinco a 10 por tratamento) foram capturadas para avaliação das concentrações plasmáticas de esteroides gonadais e composição dos ovários (biópsias e estereologia). As fêmeas foram também submetidas à uma avaliação externa relativa à espessura abdominal, o grau de rigidez abdominal e o aspecto da papila urogenital. Exceto pela ocorrência de concentrações reduzidas de 17α-hidroxiprogesterona nas FVs, durante a época de desova, observamos que FVs e FJs, mantidas em cativeiro, apresentam ciclos reprodutivos e épocas de desova similares (em dezembro para ambos os grupos). Não recomendamos o uso exclusivo da avaliação externa para a seleção de reprodutores para indução a desova, devido a uma imprecisão dos dados e falta de associação destes com a histologia e biópsia de ovário, os quais são mais precisos e confiáveis.

Equine preantral follicle harvesting, processing, and in vitro culture: the journey has already started

Preantral follicles are of great abundance in mammalian ovaries and the vast majority (>99.9%) never become ovulatory; therefore, the ability to rescue these otherwise wasted follicles seems very aooealing. Considering there are striking similarities in antral follicle dynamics between mares and women, the mare might become a good model to study early (preantral and antral) folliculogenesis in women, with several advantages related to using an animal model. Studies in our laboratory recently validated the use of a transvaginal ultrasound-guided ovarian. Biopsy Pick-Up (BPU) method to harvest preantral follicles using the mare as a model to study early folliculogenesis (Haag et al., 2013a, b, c). This article will review some of the important findings of our recent studies related to the harvesting, processing, and culture of equine preantral follicles and discuss those with the limited information availeble in the literature.

Equine preantral follicle harvesting, processing, and in vitro culture: the journey has already started

Preantral follicles are of great abundance in mammalian ovaries and the vast majority (>99.9%) never become ovulatory; therefore, the ability to rescue these otherwise wasted follicles seems very aooealing. Considering there are striking similarities in antral follicle dynamics between mares and women, the mare might become a good model to study early (preantral and antral) folliculogenesis in women, with several advantages related to using an animal model. Studies in our laboratory recently validated the use of a transvaginal ultrasound-guided ovarian. Biopsy Pick-Up (BPU) method to harvest preantral follicles using the mare as a model to study early folliculogenesis (Haag et al., 2013a, b, c). This article will review some of the important findings of our recent studies related to the harvesting, processing, and culture of equine preantral follicles and discuss those with the limited information availeble in the literature.(AU)

Effects of energy-protein supplementation frequency on performamace of primiparous grazing beef cows during pre and post-partum

O presente estudo foi realizado para avaliar os efeitos da suplementação proteicoenegética e frequência de suplementação durante o pré (105 dias antes do parto) e pósparto (105 dias após o parto) sobre o desempenho e características metabólicas de vacas primíparas de corte em pastejo. Foram utilizadas vinte e quatro vacas primíparas Nelore com PC médio de 409±8.0 kg, 22±0.9 meses de idade e 172±2.5 dias de gestação. Os tratamentos foram distribuídos aleatoriamente aos animais: Controle (mistura mineral ad libitum), Suplementação Diária (1.5 kg/dia de concentrado/animal) e Suplementação Infrequente (4.5 kg de concentrado/animal a cada três dias). O PC, GMD, ECC, esspessura de gordura subcutânea (EGS) e área de olho de lombo (AOL) foram avaliados no pré e pós-parto. Foram realizados dois ensaios de consumo e digestibilidade, o primeiro aos 55 dias antes do parto e o segundo, 55 dias após o parto. Amostras de sangue foram coletadas nos dias 27 e 9 dias antes do parto, ao parto, 9 e 27 dias após o parto para mensuração das concentrações de glicose, IGF-1, AGNE, -OH, colesterol e triglicerídeos, utilizados como indicadores do status energético, e mensuração de aminoácidos livres (AA), nitrogênio ureico sérico (NUS), proteína total, albumina e globulinas utilizados como indicadores de status proteico dos animais. As globulinas foram calculadas subtraindo a albumina quantificada do nível de proteína total. Aos 27 dias antes do parto, foi realizado biópsias de tecido hepático e tecido muscular para avaliar biomarcadores de ureogênese e metabolismo energético. Concentração sérica de progesterona (P4) foi avaliada no 36º dia pós-parto. Todas variáveis foram analisadas utilizando PROC MIXED do SAS 9.4 (SAS Inst., Cary, NC). A frequência de suplementação não alterou (P > 0.10) o PC, PC ajustado ao dia do parto (PCajus), PC após o parto (PCp), GMD, ECC, AOL e EGS no pré e pós parto. O PC (P = 0.079), PCajus (P = 0.078), e GMD (P = 0.074) foram maior para as vacas suplementadas no pré-parto. O ECC (P = 0.251), a AOL (P = 0.352), a EGS sobre o músculo longissimus (P = 0.199) e sobre o músculo Biceps femoris (P = 0.924) não foram afetados pela suplementação durante o pré-parto. Aos 105 dias após o parto, a suplementação não afetou (P > 0.10) o PC, GMD, ECC, AOL e EGS. O PC dos bezerros ao nascer, GMD e PC dos bezerros 105 dias após o parto não foram diferente (P > 0.10) de acordo com a suplementação e a frequência de suplementação. O consumo de matéria seca total e o consumo voluntário de forragem não foram afetados pela suplementação e frequência de suplementação durante o pré-parto (P > 0.10) e pósparto (P > 0.10). Os animais suplementados diariamente apresentaram maiores níveis de glicose (P < 0.001) que os animais suplementados a cada três dias. Interação entre tratamento e tempo de coleta foi significativa para AA (P < 0.001), AA glicogênicos (P < 0.001), AA cetogênicos (P < 0.001), AA glico-cetogênicos (P < 0.001) e NUS (P = 0.005). A suplementação e frequência de suplementação não alteraram (P > 0.10) os níveis séricos de IGF-1, AGNE, -OH, colesterol e triglicerídeos. Do mesmo modo, a suplementação e frequência de suplementação não alteraram (P > 0.10) os níveis de proteína total, albumina e globulinas. A abundância da proteína carbamoil-fosfato sintetase-1 (CPS-1), os níveis de mRNA de carnitina palmitoyltransferase-1 (CPT-1) e do coativador-1 do receptor ativado por proliferador de peroxissoma (PGC-1) não foram afetados (P > 0.10) pela suplementação e frequência de suplementação. Observou-se uma porcentagem semelhante (P = 0.606) de vacas com atividade ovariana (P4 > 1 ng/ml) e taxa de concepção (P = 0.842) entre os tratamentos. A suplementação proteica-energética durante o pré e pós-parto de vacas primíparas de corte em pastejo não melhora o desempenho e características metabólicas. Contudo, a redução da frequência de suplementação não resulta em efeitos negativos sobre o desempenho e características metabólicas.

The present study was performed to evaluate the effects of energy-protein supplementation and supplementation frequency during pre (105 d before calving) and postpartum (105 d after calving) of primiparous beef cows on performance and metabolic characteristics under grazing conditions. Twenty-four pregnant Nellore primiparous cows with average BW of 409±8.0 kg, 22±0.9 mo old and 172±2.5 d of gestation were used in this trial. Treatments were randomly assigned to primiparous cows: Control (ad libitum mineral mix), Daily Supplementation (1.5 kg/d of concentrate/animal) and Infrequent Supplementation (4.5 kg of concentrate/animal every three days). The BW, ADG, BCS, fat-thickness and ribeye area (RA) were evaluated at pre and postpartum period. Two 9-d intake-digestibility trials were performed throughout the experimental period where the first at 55 d before parturition, and the second at 55 d after parturition. Concentrations of glucose, IGF-1, NEFA, - OH, cholesterol and triglycerides were used as indicators of energy status, and free amino acids (AA), serum urea nitrogen (SUN), total protein, albumin and globulins were used as indicators of proteic status collected at 27 and 9 d before parturition, at the day of parturition, 9 and 27 d after parturition. Globulins were calculated subtracting the albumin quantified from the total protein level. Biopsies of hepatic and skeletal muscle tissues were performed at 27 d prior to calving to evaluate ureagenesis and energy metabolism biomarkers. Serum progesterone (P4) level was evaluated at the 36th d postpartum. The response variables were analyzed using PROC MIXED of SAS 9.4 (SAS Inst., Cary, NC). The frequency of supplementation did not alter (P > 0.10) BW, adjusted BW at day of parturition (adjBW), BW after calving upon parturition (calvingBW), ADG, BCS, RA and fat-thickness during pre and postpartum. The BW (P = 0.079), adjBW (P = 0.078) and ADG (P = 0.074) were higher for supplemented cows during the prepartum. The BCS (P = 0.251), RA (P = 0.352), fat-thickness on longissimus muscle (P = 0.199) and on Biceps femoris muscle (P = 0.924) were not affect by supplementation during the prepartum. At 105 d after calving the supplementation did not affect (P > 0.10) BW, ADG, BCS, RA, and fat-thickness. Birth BW of calves, ADG and calf BW 105 d after calving were not different (P > 0.10) according to supplementation and frequency of supplementation. The total dry matter intake and forage voluntary intake were not affected by supplementation and supplementation frequency during prepartum (P > 0.10) and postpartum (P > 0.10). Daily supplemented animals had higher (P < 0.001) glucose levels than animals supplemented every three days. Interaction between treatment and sampling time was significant for AA (P < 0.001), glucogenic AA (P < 0.001), ketogenic AA (P < 0.001), gluco/ketogenic AA (P < 0.001), and SUN (P = 0.005). The supplementation and frequency of supplementation did not alter (P > 0.10) the levels of IGF-1, NEFA, -OH, cholesterol and triglycerides. Likewise, the supplementation and frequency of supplementation did not alter (P > 0.10) the levels of total protein, albumin and globulins. The protein abundance of carbamoyl phosphate synthase-1 (CPS-1), mRNA levels of carnitine palmitoyltransferase-1 (CPT-1) and peroxisome proliferator-activated receptor coactivator-1 (PGC-1) were not affected (P > 0.10) by supplementation and supplementation frequency. Similar percentage (P = 0.606) of cows with ovarian activity (P4 > 1 ng/ml) and conception rate (P = 0.842) were observed among the treatments. The energy-protein supplementation during pre and postpartum of primiparous beef cows under grazing conditions do not improve performance and metabolic characteristics. However, the reduction of frequency of supplementation do not result negative effects on performance and metabolic characteristics.

ENSAIO CLÍNICO COM CÉLULAS-TRONCO MESENQUIMAIS (CTM) NA RECUPERAÇÃO DE OVÁRIOS DE CABRAS SUBMETIDAS A MÚLTIPLAS ASPIRAÇÕES FOLICULARES POR LAPAROSCOPIA (LOPU)

Os problemas ligados à diminuição do potencial produtivo dos ovários estão entre os principais fatores limitantes à reprodução com isso, objetivou-se com este trabalho avaliar o efeito da infusão de células-tronco mesenquimais (CTM) em ovários de cabras submetidas a múltiplas aspirações foliculares por laparoscopia (LOPU). Foram utilizadas cabras sem padrão racial definido (SPRD), não lactantes, com idade média de três anos e escore da condição corporal (ECC) variando de 3,0 a 4,0 e distribuídas em dois experimentos, no primeiro com doze cabras (n=12) e no segundo com sete (n=7). No experimento um as cabras foram sincronizadas com o uso de esponjas intravaginais com acetato de medroxiprogesterona e aplicações de d-cloprostenol e a estimulação ovariana constituiu-se na administração de FSHp e eCG, em aplicações únicas, feita 36 horas anterior às LOPUs, e posteriormente as fêmeas foram submetidas a 5 sessões/animal de LOPU com intervalos de 12 dias. No experimento dois as cabras sincronizadas da mesma forma do experimento um e posteriormente divididas em dois tratamentos: T1-CTR (Controle 3 animais); T2 - CTM (ovários tratados com células tronco mesenquimais 4 animais), incluindo a terapia celular através de infusão ovariana com células tronco CTM, na concentração de 1 x 106 cel/mL, nanomarcadas com Qtracker® e diluídas em 0,2 mL de solução fisiológica (NacL a 0,9%) em duas aplicações: na última LOPU e 70 dias após. As avaliações quanto à funcionalidade ovariana/oocitária foi realizada 45 dias posterior à ultima infusão de CTM. Coletas de sangue para dosagens hormonais de estradiol e progesterona foram realizadas, antes da primeira, e após a quinta LOPU juntamente com biopsias do córtex ovariano. A avaliação histológica foi realizada após 115 dias da primeira infusão de CTM, a partir do abate dos animais e coleta dos ovários. Todas as análises foram executadas através do programa Statistical Analysis System (SAS Institute Inc., 2002). No experimento um, houve diferença significativa (P<0,05) na média de folículos aspirados com menor média na 3ª LOPU. Em relação à média de oócitos recuperados, não observou-se diferença significativa entre a 1ª e a 5ª LOPU (P>0,05), porem as médias dos folículos aspirados na 2ª, 3ª e 4ª LOPU reduziram significativamente em relação a 1ª LOPU (P<0,05). Quanto a taxa de recuperação oocitária, houve uma redução na 3ª e 4ª LOPU em relação à 1ª e 2ª LOPU (P<0,05) e na 5ª LOPU observou-se um aumento da recuperação oocitaria em relação à 3ª e 4ª LOPU (P<0,05). Quanto a qualidade, os CCOs de grau 2 reduziram entre a 1ª e a 4ª LOPU e os oócitos de grau 4 aumentaram na 2ª, 4ª e 5ª LOPU em relação a 1ª e 3ª LOPU(P<0,05). Houve uma redução (P<0,05) de estruturas viáveis na 2ª e 4ª LOPU (46,4% e 53,8% respectivamente), com retomada da viabilidade na 3ª e 5ª LOPU (80,76% e 61,2% respectivamente). As biópsias que antecederam as coletas e pós sucessivas aspirações evidenciaram tecidos ovarianos morfologicamente normais, com alterações discretas no parênquima ovariano. Não houve diferença (P>0,05) na dosagem de progesterona entre as sucessivas LOPUs porem houve um aumento (P<0,05) na dosagem de estradiol da 2º LOPU em relação às demais. No experimento 2, não houve diferença significativa (P>0,05) entre os folículos aspirados, oócitos recuperados e taxa de recuperação em relação aos grupos controle e grupos tratados com CTMs. Em relação a qualidade dos CCOs não ocorreu diferença significativa (P<0,05) entre as classificações das estruturas oocitárias. Nas avaliações histológicas foram observadas discretas alterações no parênquima ovariano, sem comprometimento do mesmo. Conclui-se que, cinco sessões de aspirações foliculares por laparoscopia com intervalos de doses dias não interfere morfofuncionalmente nos ovários de cabras e que a utilização de CTM não influenciou nos parâmetros qualitativos e quantitativos de oócitos recuperados por LOPU de cabras SPRD.

The problems related to the reduction of ovaries productive potential are among the main limiting factors for reproduction, therefore, the objective of this work was to evaluate the effect of the infusion of mesenchymal stem cells (CTM) on ovary of goats submitted to multiple follicular aspirations by laparoscopy (LOPU). Goats with undefined racial pattern (SPRD), non-lactating, with mean age of three years and body condition scoring (ECC) ranging from 3.0 to 4.0 were used and distributed in two experiments, in the first with twelve goats (n= 12) and in the second with seven (n = 7). In the first experiment, the goats were synchronized using intravaginal sponges with medroxyprogesterone acetate and d-cloprostenol applications, ovarian stimulation consisted of administration of FSHp and eCG in single applications 36 hours prior to LOPUs, and later females were submitted to 5 sessions/animal of LOPU at 12-day intervals. In the second experiment, the goats were synchronized in the same way of the first experiment and later divided into two treatments: T1-CTR (Control - 3 animals); T2-CTM (ovaries treated with mesenchymal stem cells - 4 animals), including cell therapy through ovarian infusion in two applications with 1 x 106 cel/mL of stem cells - CTM, nanoparticulated with Qtracker ® and diluted in 0.2 mL of physiological solution (0.9% NaCl), these two applications were performed at the last LOPU and 70 days after. Evaluations for ovarian / oocyte functionality were performed 45 days after the last CTM infusion. Blood collections for hormonal dosages of estradiol and progesterone were performed, before the first LOPU, and after the fifth LOPU simultaneously to biopsies of the ovarian cortex. The histological evaluation was performed after the animals slaughter and collection of the ovaries, 115 days after the first CTM infusion. All analyzes were performed through the Statistical Analysis System (SAS Institute Inc., 2002). In the first experiment, there was a significant difference (P <0.05) in the mean of the aspirated follicles with lower mean in the 3rd LOPU. Regarding the mean of oocytes recovered, no significant difference was observed between the 1st and 5th LOPU (P> 0.05), however the mean of follicles aspirated in 2nd, 3rd and 4th LOPU significantly reduced compared to 1st LOPU (P <0.05). Regarding the oocyte recovery rate, there was a reduction in the 3rd and 4th LOPU in relation to the 1st and 2nd LOPU (P <0.05) and in the 5th LOPU there was an increase in oocyte recovery in relation to the 3rd and 4th LOPU (P <0.05). As regards the quality, the grade 2 levels of the CCO system increased between 1st and 4th LOPU and grade 4 levels increased in 2nd, 4th and 5th LOPU in relation to 1st and 3rd LOPU (P <0.05). There was a reduction (P <0.05) of viable structures in the 2nd and 4th LOPU (46.4% and 53.8%, respectively), with recovery of viability in the 3rd and 5th LOPU (80.76% and 61.2% respectively). The biopsies that preceded the collections and subsequent successive aspirations revealed morphologically normal ovarian tissues, with discrete alterations in the ovarian parenchyma. There was no difference (P> 0.05) in the progesterone dosage between successive LOPU's, but there was an increase (P <0.05) in the estradiol dosage of the 2nd LOPU in relation to the others. In the second experiment, there was no significant difference (P> 0.05) between aspirated follicles, recovered oocytes and recovery rate in relation to the control groups and groups treated with CTM's. Regarding the quality of CCOs, there was no significant difference (P <0.05) between the classifications of oocyte structures. In the histological evaluations, discrete alterations in the ovarian parenchyma were observed, without any impairment. It was concluded that five sessions of follicular aspiration by laparoscopy with dose and day intervals did not interfere morphofunctionally in the ovaries of goats and that the use of CTM did not influence the qualitative and quantitative parameters of oocytes recovered by LOPU from SPRD goats.

Níveis de prostaglandina F durante a desova induzida de Pacu (Piaractus mesopotamicus).

O objetivo deste trabalho foi determinar os níveis plasmáticos de prostaglandina F (PGF) de matrizes de pacu (Piaractus mesopotamicus) ao longo do período de reprodução induzida por hipofisação. Para isso, oito fêmeas foram induzidas com extrato bruto de hipófise de carpa e oito fêmeas foram injetadas apenas com solução salina (grupo controle). Amostras de sangue e biópsias de ovário foram coletadas respectivamente para determinação das concentrações plasmáticas de PGF e determinação dos percentuais de diferentes tipos de ovócitos nos seguintes períodos: primeira dose hormonal, segunda dose hormonal e 2h, 4h, 6h e 8h (período da ovulação) após a segunda dose hormonal. Os peixes foram separados em três grupos: controle (C), tratadas que desovaram (TD) e tratadas que não desovaram (TN). Ovócitos com quebra da vesícula germinativa (GVBD) foram observados pela primeira vez 4 horas após a segunda dose hormonal nas fêmeas TD e TN. Não foram encontrados ovócitos GVBD no grupo C. A concentração de PGF do grupo TD foi superior aos outros grupos seis horas após a segunda dose. Oito horas após a segunda dose a concentração de plasmática de PGF dos grupos de TD e TN foram similares entre si, mas superiores ao grupo C. Concluímos que os valores superiores de PGF das fêmeas TD, 6 horas após a segunda dose, indicam que a elevação nos níveis de PGF neste período pode estar associada com a desova bem-sucedida na espécie, especialmente por se conhecer o potencial papel indutor da ovulação em peixes desta substância. Assim, por meio de uso de PGF exógena, pretendemos, em futuras abordagens, provocar um pico artificial desta substancia 6 horas após segunda dose visando avaliar a possibilidade de se obter maiores taxas de ovulação nesta espécie.

The objective of this work was to determine the plasma levels of prostaglandin F (PGF) from pacu matrices (Piaractus mesopotamicus) throughout the reproduction period induced by hypophysis. For this, eight females were induced with crude carp pituitary extract and eight females were injected with saline solution only (control group). Blood samples and ovarian biopsies were collected for the determination of plasma concentrations of PGF and percentages of different oocyte types in the following periods: first hormonal dose, second hormonal dose and 2h, 4h, 6h, and 8h (ovulation period) after the second hormonal dose. The fish were separated into three groups: control (C), treated that spawned (TD) and treated that did not spawn (TN). Germ cell ruptured oocytes (GVBD) were first observed 4 hours after the second hormonal dose in TD and TN females. No GVBD oocytes were found in-group C. The concentration of PGF in the TD group was higher than the other groups six hours after the second dose. Eight hours after the second dose the PGF plasma concentration of the TD and TN groups were similar to each other, but higher than the C group. We concluded that the higher PGF values of the TD females, 6 hours after the second dose, indicate that the elevation in PGF levels in this period may be associated with successful spawning in the species, especially since the potential role of ovulation in fish of this substance is known. Thus, using exogenous PGF, we intend, in future approaches, to elicit an artificial peak of this substance 6 hours after the second dose in order to evaluate the possibility of obtaining higher rates of ovulation in this species.

Detecção imunoistoquímica de receptores de estrógeno e progesterona no endométrio de vacas Nelore (Bos taurus indicus) durante o anestro pós-parto / Immunohistochemical detection of receptors for estrogen and progesterone in endometrial glands and stroma during the pospartum anestrus in Nelore (Bos taurus indicus) cows

Foram utilizadas 24 vacas Nelore P.O., em anestro pós-parto, diagnosticado pelo histórico reprodutivo, por avaliações ultrassonográficas transretais e por dosagem de progesterona plasmática, que foram submetidas à colheita de fragmento uterino via transcervical. Os animais foram divididos em dois grupos conforme o máximo diâmetro folicular: grupo 1: folículos <6mm (n=12); grupo 2: folículos >6mm (n=12). Para avaliar receptor de estrógeno e receptor de progesterona no epitélio glandular e no estroma, foi utilizada a técnica de imunoistoquímica. Altas contagens relativas e alta intensidade de marcação para receptor de estrógeno e progesterona no epitélio glandular e no estroma foram observadas nos dois grupos. No entanto, a intensidade de marcação para o receptor de progesterona no epitélio glandular foi mais alta no grupo 2 comparado ao grupo 1. Quando o epitélio glandular e o estroma foram comparados, o número relativo de receptor de estrógeno no grupo 1 foi mais alto no epitélio glandular comparado ao estroma, e a intensidade de marcação para o receptor de progesterona no grupo 2 foi mais alta no epitélio glandular comparado ao estroma. Os resultados sugerem que os mecanismos que controlam a expressão de receptores no anestro são semelhantes aos observados durante o ciclo estral.(AU)

Twenty-four postpartum anestrous Nelore purebred cows were used in the study. Anestrous was determined based on the reproductive history which was confirmed in each cow based on plasma progesterone concentration and by transrectal ultrasonography. Endometrial biopsies were collected. The animals were separated into two groups according to maximum follicular diameter- Group 1: follicles <6mm (n=12) and Group 2: follicles >-6mm follicles (n=12). The immunohistochemistry technique was employed to evaluate the presence of estrogen and progesterone receptors in the uterine glandular epithelium and stroma. High counts of positive nuclei and high intensity of immunostain for estrogen and progesterone receptors in the glandular epithelium and stroma were observed in the two groups. However, the immunostain intensity of progesterone receptors in the glandular epithelium was higher in Group 2 compared to Group 1. When glandular epithelium and stroma were compared within each group, the relative number of estrogen receptors in the Group 1 was higher in the glandular epithelium compared to stroma and the immunostain intensity for the progesterone receptor in Group 2 was higher in the glandular epithelium compared to stroma. The results suggest that the mechanisms that control the expression of endomerial receptors in the anestrus are similar to the ones observed during the estrus cycle.(AU)

Avaliação ultrassonográfica torácica em cães com efusão pleural e/ou periocárdica / Thoracic ultrasonographyc evaluation of dogs with pleural and/or pericardial effusion

Com o objetivo de avaliar a ultrassonografia torácica (incluindo a ecocardiografia) como método de exame complementar em pacientes portadores de efusão pleural e/ou pericárdica, realizou-se a ultrassonografia torácica em 30 cães, machos e fêmeas de raças e idades variadas. Animais atendidos nos serviços de Pronto-Atendimento, Clínica Médica ou Clínica Cirúrgica do HOVET/USP com efusão pleural e/ou pericárdica, detectadas por meio de exame radiográfico, ultrassonografia emergencial ou toracocentese exploratória foram incluídos no estudo. Observou-se efusão pleural em 12 cães (40%), efusão pericárdica em oito (26,66%) e efusão pleural e pericárdica em outros dez cães (33,33%). A causa da efusão pleural e/ou pericárdica foram avaliadas ultrassonograficamente como sendo: nódulo/tumor cardíaco (5 - 16,66%), nódulo/tumor intratorácico (5 16,66%), insuficiência cardíaca congestiva por cardiomiopatia dilatada (4 13,33%) ou endocardiose de mitral e tricúspide (3 10%), efusão pericárdica idiopática (3 - 10%), li fossarcoma (2 - 6,66%), piotórax (2 6,66%), ruptura diafragmática (1 3,33%), hérnia peritônio-pericárdica (1 –3,33%), pneumonia e pleurite (1 - 3,33%), tumor de ovário com metástases torácicas (1 3,33%), pericardite infecciosa (cinomose) (1 3,33%) e hipoalbuminemia (1 3,33%).(AU)

To evaluate diagnostic accuracy of thoracic ultrasonography (including echocardiography) of patients with pleural and/or pericardial effusion, thoracic ultrasonography was performed in 30 dogs, males and females, of different breeds and ages. The animals were admitted to the Emergency, Internal medicine or Surgery department of the Faculty of Veterinary Medicine, University of São Paulo, presenting with pleural and/or pericardial effusion, diagnosed by thoracic radiography, emergency thoracic ultrasonography or exploratory thoracocentesis. Twelve (40%) dogs had pleural effusion, 9 (30%) had pericardial effusion and 9 (30%) had both pleural and pericardial effusions. The definitive cause of effusion were obtained by thoracic ultrasonography as follows: heart mass (5 –16,66%), intrathoracic mass (5 16,66%), congestive heart failure by dilated cardiomyopathy (4 13,33%) or mitral and tricuspid insufficiency (3 10%), idiopathic pericardial effusion (3 10%), lymphosarcoma (2 6,66%), pyothorax (2 - 6,66%), traumatic diaphragmatic hernia (1 3,33%), congenital peritoneopericardial hernia (1 3,33%), pneumonia and pleuritis (1 3,33%), ovarian neoplasia and thoracic metastasis (1 3,33%), infectious pericarditis (distemper) (1 3,33%) and hypoalbuminemia (1 3,33%). The conclusion was that thoracic ultrasonography was an excellent auxiliary exam in animals with pleural and/or pericardial effusion, and its not invasive and safe for the patient, allowing to guide biopsies and perform the exam in different decubits, avoiding patient stress.(AU)

Biopsias luteais em vacas nelore / Luteal biosy in nellore cows

O presente estudo descreve uma técnica para obtenção de fragmentos luteais empregando-se a colpotomia. Os animais foram submetidos à anestesia epidural e no fórnix vaginal e, após dez minutos, realizou-se uma incisão no fundo vaginal com lâmina de bisturi, bem como procedeu-se à dissecção do tecido, até que fossem possíveis o acesso à cavidade pélvica e a tração dos ovários para o interior vaginal. Realizou-se, então, a colheita de biopsia luteínica, com o auxílio de uma pinça do tipo Yomann. Sinais de dor e estresse foram observados apenas em duas colheitas durante a incisão no fundo vaginal, a tração do ovário ou durante a biopsia luteínica. Contudo, esses sinais foram observados em dez colheitas durante a dissecção da parede vaginal e peritôneo. Registrou-se a ocorrência de ataxia em 38,81% colheitas, a qual esteve relacionada, normalmente, a um procedimento mais longo. As ataxias podem ser divididas em leve (15/26), moderada (6/26) e severa (5/26). A avaliação da presença de adesões ovarianas ipsilaterais à incisão realizou-se apenas até a quarta colheita, sendo notadas em dezesseis colheitas. O protocolo empregado mostrou-se um método seguro e eficiente na obtenção de fragmentos luteais. A baixa incidência de aderências permite o uso consecutivo das fêmeas sem interferência nas ovulações e colheitas subsequentes.

The present study describes a technique to obtain consecutive luteal samples by colpotomy. The animals received an epidural anesthesia and local anesthesia (vaginal vault) and after ten minutes the vaginal vault was incised with a scalpel blade and tissue was dissected to provide access to the pelvic cavity and to retract the ovary into the vagina. Then, a luteal biopsy was performed with a Yomann biopsy nipper. Signs indicative of pain and stress during the vaginal vault incision, traction of ovary or luteal biopsy were observed only in two collections. However, these signs were observed in ten collections during dissection of the vaginal wall and peritoneum. The occurrence of ataxia was observed in 26 collections and it was usually related to a longer duration of the procedure. Ataxia could be divided in light (15/26), moderate (6/26) and severe (5/26). The occurrence of ovarian adhesions ipsilateral to the incision was evaluated only in the initial four collections. Adhesions were present in 16 collections. The protocol described above provided a safe and efficient method to acquire luteal samples. The low incidence of adhesions allows the consecutive use of females without any interference in subsequent ovulations and collections.

Biopsias luteais em vacas nelore / Luteal biosy in nellore cows

O presente estudo descreve uma técnica para obtenção de fragmentos luteais empregando-se a colpotomia. Os animais foram submetidos à anestesia epidural e no fórnix vaginal e, após dez minutos, realizou-se uma incisão no fundo vaginal com lâmina de bisturi, bem como procedeu-se à dissecção do tecido, até que fossem possíveis o acesso à cavidade pélvica e a tração dos ovários para o interior vaginal. Realizou-se, então, a colheita de biopsia luteínica, com o auxílio de uma pinça do tipo Yomann. Sinais de dor e estresse foram observados apenas em duas colheitas durante a incisão no fundo vaginal, a tração do ovário ou durante a biopsia luteínica. Contudo, esses sinais foram observados em dez colheitas durante a dissecção da parede vaginal e peritôneo. Registrou-se a ocorrência de ataxia em 38,81% colheitas, a qual esteve relacionada, normalmente, a um procedimento mais longo. As ataxias podem ser divididas em leve (15/26), moderada (6/26) e severa (5/26). A avaliação da presença de adesões ovarianas ipsilaterais à incisão realizou-se apenas até a quarta colheita, sendo notadas em dezesseis colheitas. O protocolo empregado mostrou-se um método seguro e eficiente na obtenção de fragmentos luteais. A baixa incidência de aderências permite o uso consecutivo das fêmeas sem interferência nas ovulações e colheitas subsequentes.(AU)

The present study describes a technique to obtain consecutive luteal samples by colpotomy. The animals received an epidural anesthesia and local anesthesia (vaginal vault) and after ten minutes the vaginal vault was incised with a scalpel blade and tissue was dissected to provide access to the pelvic cavity and to retract the ovary into the vagina. Then, a luteal biopsy was performed with a Yomann biopsy nipper. Signs indicative of pain and stress during the vaginal vault incision, traction of ovary or luteal biopsy were observed only in two collections. However, these signs were observed in ten collections during dissection of the vaginal wall and peritoneum. The occurrence of ataxia was observed in 26 collections and it was usually related to a longer duration of the procedure. Ataxia could be divided in light (15/26), moderate (6/26) and severe (5/26). The occurrence of ovarian adhesions ipsilateral to the incision was evaluated only in the initial four collections. Adhesions were present in 16 collections. The protocol described above provided a safe and efficient method to acquire luteal samples. The low incidence of adhesions allows the consecutive use of females without any interference in subsequent ovulations and collections.(AU)

Induction of oocyte maturation in the white croaker Micropogonias furnieri (Pisces: Sciaenidae) by human chorionic gonadotropin

The present work aimed to identify the best doses of human chorionic gonadotropin (hCG) needed to induce oocyte maturation of Micropogonias furnieri and to characterize ovarian dynamics during the periovulatory period. Adult M. furnieri females with fully developed ovaries were injected intraperitoneally with four different doses of hCG. The gonadotropin response was suceeded by analyzing morphologically gonadal biopsies and following the postinjection changes in follicle diameter. Oocyte maturation was induced by three doses used: 100, 300, and 500 IU of hCG kg bw-1, and was reached 48 h after treatment with 300 and 500 IU of hCG kg bw-1, and 72 h after treatment with 100 IU of hCG kg bw-1. Concerning ovarian dynamics, only 100 and 300 IU of hCG kg bw-1 mimicked the natural ones which have a synchronic group maturation. In conclusion, the dose mimicking natural ovarian dynamics and inducing oocyte maturation more quickly is 300 IU of hCG kg bw-1.

O presente estudo teve por objetivo identificar as melhores doses de gonadotrofina cariônica humana (hCG) necessárias para maturação de oócitos de Micropogonias furnieri e caracterizar a dinâmica do ovário ao longo do período pré-ovulatório. Fêmeas adultas de M. furnieri, com ovários completamente desenvolvidos, receberam intraperitonealmente quatro doses diferentes de hCG. A resposta à gonadotrofina foi acompanhada de análise morfológica de biópsias gonadais e mudanças no diâmetro dos folículos após estimulação. A maturação dos oócitos foi induzida por três doses de hCG kg bw-1, 100, 300 e 500 UI. A maturação dos oócitos foi atingida após 48 h de tratamento com 300 e 500 UI de hCG kg bw-1 e após 72 h de tratamento com 100 UI de hCG kg bw-1. Em relação à dinâmica do ovário, apenas os tratamentos com 100 e 300 UI de hCG kg bw-1 reproduziram sua dinâmica natural, apresentando maturação em grupo sincronizada. Esses resultados permitem concluir que a dose capaz de reproduzir a dinâmica do ovário e induzir maturação dos oócitos em curtos períodos é de 300 UI de hCG kg bw-1.

Vitrification of Ovarian Tissue from Non-Human Primates

Background: One of the strategies to preserve genetic material from nonhuman primates (NHP) consists in the implementation of germplasm banks, for future application in reproductive biotechniques, as well as for biomedical research. Based on the success rates achieved in human, there is a prominent possibility to succeed also with NHP. However, studies with NHP are still scarce, especially regarding the cryopreservation of ovarian tissue.Review: Neotropical non-human primates, especially males, have been used in research related to reproductive biotechniques in Brazil. Regarding research on female reproduction and ovarian tissue preservation, most studies were performed using domestic animals as models. Current concepts and controversies in the restoration of gametes in adult females does not exclude the needs to preserve ovarian tissue. Importantly, ovarian tissue can be collected and preserved even after the death of the donors, being applied when finding dead females. Furthermore, collection of ovarian biopsies is also feasible and will not affect reproductive function. Among the cryopreservation methods, the vitrification has been indicated due to practical logistic, as well as because it will avoid the formation of large intracellular ice crystals, and it is claimed that ovarian stromal damage will be decreased under vitrification. Considering the number of threatened prim