Curcumin alleviated oxidation stress injury by mediating osteopontin in nephrolithiasis rats

Purpose: To explore the role and mechanism of curcumin (Cur) in reducing oxidative stress damage in rats with nephrolithiasis induced by ethylene glycol (EG). Methods: Thirty male rats were divided into normal control, model, positive (10% potassium citrate), Cur-10 (10 mg/kg curcumin) and Cur-20 (20 mg/kg curcumin) groups. Results: The results of kidney tissue section stained by hematoxylin-eosin and von Kossa showed that curcumin treatment can inhibit the formation of kidney stones. The biochemical test results showed that the urea (Ur), creatinine (Cr), uric acid (UA), inorganic phosphorus and Ca2+ concentrations in urine decreased after being treated with curcumin. There were significant differences between different doses of curcumin (P < 0.05). Compared with the Cur-10 group, Cur-20 had a more significant inhibitory effect on malondialdehyde (MDA) (P < 0.05). In addition, reverse transcription polymerase chain reaction (PCR) detection and immunohistochemical results indicated that the osteopontin (OPN) in the kidney was significantly reduced after curcumin treatment. Conclusion: Curcumin could reduce the oxidative stress damage caused by EG-induced kidney stones.

Utilização da manufatura aditiva (impressão em 3D) no tratamento de osteossarcoma vertebral em cão submetido a vertebrectomia lombar: relato de caso / Use of additive manufacturing (3D printing) in the treatment of vertebral osteosarcoma in dogs submitted to lumbar vertebrectomy: case report / Uso de la fabricación aditiva (impresión 3D) en el tratamiento del osteosarcoma vertebral en perros sometidos a vertebrectomía lumbar: caso clínico

O osteossarcoma é uma das neoplasias ósseas mais comumente relatadas na medicina veterinária, principalmente em cães de raça grande e gigante, sendo que a forma vertebral possui importante cenário clínico apresentando 5% do total de relatos. Este possui um prognóstico desfavorável e tempo de sobrevida pequeno. A apresentação clínica é variável e podem ser observados sinais de dor intensa a déficits neurológicos em decorrência do envolvimento medular progressivo. O diagnóstico pode ser realizado por imagens através de radiografia ou tomografia computadorizada, e por análises laboratoriais de citopatologia ou histopatologia, sendo o último preferível para classificação de tumores. A ressecção tumoral parcial é uma forma paliativa de tratamento, indicada em alguns casos para aliviar temporariamente os sinais clínicos relacionados a dor e déficits neurológicos. Contudo, quando há possibilidade terapêutica, a excisão completa fornece melhor prognóstico e tempo de sobrevida, principalmente quando associada a quimioterapia ou radioterapia. Mediante procedimento de vertebrectomia, a estabilização cirúrgica é necessária, podendo ser realizada respeitando os princípios para estabilização vertebral por trauma, sendo necessária a colocação de um espaçador vertebral para evitar o colabamento medular. Diversos materiais foram propostos para a confecção do espaçador vertebral. A manufatura aditiva, também conhecida como impressão 3D, tem transformado o conceito de prototipagem rápida em realidade, devido a habilidade de fabricar peças geométricas específicas de alta complexidade e de forma rápida, permitindo elaborar protótipos para uso pré ou transoperatórios em cirurgias ortopédicas de alta complexidade. O polietileno tereftalato glicol (PETG), por ser um termoplástico de alta resistência, não-biodegradável, biocompatível e de baixo custo é foco desse trabalho no desenvolvimento do espaçador vertebral como estabilizador em um procedimento de vertebrectomia em cão.

Osteosarcoma is one of the most commonly reported bone neoplasms in veterinary medicine, especially in large and giant breed dogs, and the vertebral form has an important clinical scenario presenting 5% of the total reports. This has an unfavorable prognosis and a short survival time. The clinical presentation is variable and signs of severe pain to neurological deficits can be observed as a result of progressive spinal cord involvement. The diagnosis can be made by imaging with radiography or computed tomography, and by laboratory analysis of cytopathology or histopathology, the latter being preferable for tumor classification. Partial tumor resection is a palliative form of treatment, indicated in some cases to temporarily relieve clinical signs related to pain and neurological deficits. However, when there is therapeutic possibility, complete excision provides better prognosis and survival time, especially when associated with chemotherapy or radiotherapy. Through the vertebrectomy procedure, surgical stabilization is necessary and can be performed following the principles for vertebral stabilization due to trauma, with the placement of a vertebral spacer being necessary to prevent spinal collapse. Several materials have been proposed for the manufacture of the spinal spacer. Additive manufacturing, also known as 3D printing, has transformed the concept of rapid prototyping into reality, due to its ability to quickly manufacture specific geometric parts of high complexity, allowing the elaboration of prototypes for pre- or trans-operative use in highly complex orthopedic surgeries. Polyethylene terephthalate glycol (PETG), being a high-strength, non-biodegradable, biocompatible and low-cost thermoplastic, is the focus of this work in the development of the vertebral spacer as a stabilizer in a vertebrectomy procedure in a dog.

El osteosarcoma es una de las neoplasias óseas más comúnmente reportadas en medicina veterinaria, principalmente en perros de raza grande y gigante, siendo que la forma vertebral tiene importante escenario clínico presentando el 5% del total de reportes. Tiene un pronóstico desfavorable y un tiempo de supervivencia corto. La presentación clínica es variable y pueden observarse desde signos de dolor intenso hasta déficits neurológicos como resultado de la afectación medular progresiva. El diagnóstico puede realizarse mediante imágenes con radiografía o tomografía computarizada, y mediante análisis de laboratorio de citopatología o histopatología, siendo este último preferible para la clasificación del tumor. La resección parcial del tumor es una forma de tratamiento paliativo, indicada en algunos casos para aliviar temporalmente los signos clínicos relacionados con el dolor y los déficits neurológicos. Sin embargo, cuando hay posibilidad terapéutica, la escisión completa proporciona un mejor pronóstico y tiempo de supervivencia, principalmente cuando se asocia con quimioterapia o radioterapia. Mediante el procedimiento de la vertebrectomía, es necesaria la estabilización quirúrgica, que puede realizarse respetando los principios para la estabilización vertebral por traumatismo, siendo necesaria la colocación de un espaciador vertebral para evitar el colapso medular. Se han propuesto varios materiales para la fabricación del espaciador vertebral. La fabricación aditiva, también conocida como impresión 3D, ha transformado el concepto de prototipado rápido en una realidad, debido a la capacidad de fabricar piezas geométricas específicas de gran complejidad y con rapidez, lo que permite la elaboración de prototipos para su uso pre o trans-operatorio en cirugías ortopédicas de gran complejidad. El polietileno tereftalato glicol (PETG), por ser un termoplástico de alta resistencia, no biodegradable, biocompatible y de bajo costo es el foco de este trabajo en el desarrollo del espaciador vertebral como estabilizador en un procedimiento de vertebrectomía en un perro.

The protective effect of wild mustard (Sinapis arvensis L) pollen seeds against the toxicity of a solvent (EGME) in Wistar rats

Environmental pollution is the unfavorable alteration of our environment, wholly or mainly as a by-product of human actions, by the direct or indirect effects of changes in energy patterns, radiation levels, the chemical and physical constitution, and the abundance of the organisms. Thus, the environment's pollution is one of the most severe problems humanity and other forms of life face today on our planet, and this population makes severe disruption. The xenobiotic substances responsible for this pollution are numerous and diverse due to multiple human activities that can be the source. However, attention mainly focuses on fertilizers, pesticides, heavy metals, and certain petrochemicals of significant consumption, such as solvents. The purpose of this study is to highlight the reprotoxic and hepatotoxic effects of Ethylene Glycol Monoethyl Ether (EGME) on male Wistar rats. Male Wistar rats were exposed to Ethylene Glycol Monomethyl Etheralone (EGME) (500 mg/kg), combined with the aqueous extract of the pollen grains of Sinapis arvensis (P-EGME) (300mg/kg), in addition to the control (T) and positive control (P) groups for a period of 4 weeks by gavage, to estimate the protective role of this plant against the intoxication of EGME. The results show that EGME can induce reprotoxic effects revealed by a reduction in testes and epididymis mass accompanied by decreased male fertility indicators (sperm concentration). The results also show that treatment with EGME caused a reduction in the red blood cell number, hemoglobin and hematocrit rate, white blood cells in the treated groups compared to the control groups. The present study revealed that treatment with Ethylene Glycol Monoethyl Ether (EGME) under the same experimental conditions could affect several biological markers, especially male fertility. Besides, the protective activity of the wild mustard (Sinapis arvensis) pollen in the face of cellular [...] (AU)

The protective effect of wild mustard (Sinapis arvensis L) pollen seeds against the toxicity of a solvent (EGME) in Wistar rats

Environmental pollution is the unfavorable alteration of our environment, wholly or mainly as a by-product of human actions, by the direct or indirect effects of changes in energy patterns, radiation levels, the chemical and physical constitution, and the abundance of the organisms. Thus, the environment's pollution is one of the most severe problems humanity and other forms of life face today on our planet, and this population makes severe disruption. The xenobiotic substances responsible for this pollution are numerous and diverse due to multiple human activities that can be the source. However, attention mainly focuses on fertilizers, pesticides, heavy metals, and certain petrochemicals of significant consumption, such as solvents. The purpose of this study is to highlight the reprotoxic and hepatotoxic effects of Ethylene Glycol Monoethyl Ether (EGME) on male Wistar rats. Male Wistar rats were exposed to Ethylene Glycol Monomethyl Etheralone (EGME) (500 mg/kg), combined with the aqueous extract of the pollen grains of Sinapis arvensis (P-EGME) (300mg/kg), in addition to the control (T) and positive control (P) groups for a period of 4 weeks by gavage, to estimate the protective role of this plant against the intoxication of EGME. The results show that EGME can induce reprotoxic effects revealed by a reduction in testes and epididymis mass accompanied by decreased male fertility indicators (sperm concentration). The results also show that treatment with EGME caused a reduction in the red blood cell number, hemoglobin and hematocrit rate, white blood cells in the treated groups compared to the control groups. The present study revealed that treatment with Ethylene Glycol Monoethyl Ether (EGME) under the same experimental conditions could affect several biological markers, especially male fertility. Besides, the protective activity of the wild mustard (Sinapis arvensis) pollen in the face of cellular [...]

Effect of cryoprotectant on the motility, viability, fertilization, and DNA integrity of naleh fish Barbonymus sp. (Cyprinidae) sperm / Efeito do crioprotetor na motilidade, viabilidade, fertilização e integridade do DNA no sêmen de peixes naleh Barbonymus sp. (Cyprinidae)

Naleh fish Barbonymus sp. is a commercial freshwater fish, which is indigenous to Aceh, Indonesia. The population of this species has declined over the years as a result of habitat perturbations and overfishing. Hence, the crucial need to develop a cryopreservation method to support breeding programs. This involved the use of a cryoprotectant as an important component. The objective of this study, therefore, was to explore the best cryoprotectant for naleh fish spermatozoa, and a total of five types were tested. These include the DMSO, Methanol, Ethanol, Glycerol, and Ethylene Glycol at a similar concentration of 10%, which were individually combined with 15% egg yolk, and every treatment was performed in three replications. Conversely, Ringer's solution was adopted as an extender, and the sperm was cryopreserved in liquid nitrogen for 15 days. The results showed significant influence on sperm motility and viability, as well as egg fertility of naleh fish (P <0.05), although the DMSO provided the best outcome, compared to others at 47.17%, 50.13%, and 45.67%, respectively. Furthermore, DNA fragmentation had not occurred in the fresh and cryopreserved sperm samples, indicating the protective effect of tested cryoprotectants. It is concluded that the 10% DMSO and 15% egg yolk is the best cryoprotectant for naleh fish spermatozoa.(AU)

O peixe naleh Barbonymus sp. é um peixe comercial de água doce, originário de Aceh, Indonésia. Durante vários anos, as perturbações provocadas no seu habitat e a pesca predatória determinaram o declínio da sua população, cuja preservação deve apoiar-se em um programa de reprodução controlada, com o emprego de espermatozoides criopreservados. O presente trabalho realizou um estudo comparativo de cinco crioprotetores: dimetilsultóxido, metanol, etanol, glicerol e etileno glicol. Todos os crioprotetores foram testados na concentração de 10%, combinados a 15% de gema de ovo. Cada tratamento foi efetuado em triplicatas. A solução de ringer foi utilizada como extensor e o esperma foi criopreservado em nitrogênio líquido por 15 dias. Os resultados obtidos revelaram a existência de influência significante (P<0,05) na viabilidade e motilidade espermática bem como na fertilidade dos ovos do peixe naleh, em que o dimetilsulfóxido apresentou o melhor resultado com os valores de 47,17%, 50,13% e 45,67%, respectivamente. Por outro lado, a fragmentação do DNA não ocorreu nas amostras de esperma fresco e criopreservado, indicando o efeito protetor dos crioprotetores testados. A conclusão obtida foi que o dimetilsulfóxido e 15% de gema de ovo foram o melhor crioprotetor para os espermatozoides do peixe naleh.(AU)

Effect of cryoprotectant on the motility, viability, fertilization, and DNA integrity of naleh fish Barbonymus sp. (Cyprinidae) sperm / Efeito do crioprotetor na motilidade, viabilidade, fertilização e integridade do DNA no sêmen de peixes naleh Barbonymus sp. (Cyprinidae)

Naleh fish Barbonymus sp. is a commercial freshwater fish, which is indigenous to Aceh, Indonesia. The population of this species has declined over the years as a result of habitat perturbations and overfishing. Hence, the crucial need to develop a cryopreservation method to support breeding programs. This involved the use of a cryoprotectant as an important component. The objective of this study, therefore, was to explore the best cryoprotectant for naleh fish spermatozoa, and a total of five types were tested. These include the DMSO, Methanol, Ethanol, Glycerol, and Ethylene Glycol at a similar concentration of 10%, which were individually combined with 15% egg yolk, and every treatment was performed in three replications. Conversely, Ringer's solution was adopted as an extender, and the sperm was cryopreserved in liquid nitrogen for 15 days. The results showed significant influence on sperm motility and viability, as well as egg fertility of naleh fish (P <0.05), although the DMSO provided the best outcome, compared to others at 47.17%, 50.13%, and 45.67%, respectively. Furthermore, DNA fragmentation had not occurred in the fresh and cryopreserved sperm samples, indicating the protective effect of tested cryoprotectants. It is concluded that the 10% DMSO and 15% egg yolk is the best cryoprotectant for naleh fish spermatozoa.(AU)

O peixe naleh Barbonymus sp. é um peixe comercial de água doce, originário de Aceh, Indonésia. Durante vários anos, as perturbações provocadas no seu habitat e a pesca predatória determinaram o declínio da sua população, cuja preservação deve apoiar-se em um programa de reprodução controlada, com o emprego de espermatozoides criopreservados. O presente trabalho realizou um estudo comparativo de cinco crioprotetores: dimetilsultóxido, metanol, etanol, glicerol e etileno glicol. Todos os crioprotetores foram testados na concentração de 10%, combinados a 15% de gema de ovo. Cada tratamento foi efetuado em triplicatas. A solução de ringer foi utilizada como extensor e o esperma foi criopreservado em nitrogênio líquido por 15 dias. Os resultados obtidos revelaram a existência de influência significante (P<0,05) na viabilidade e motilidade espermática bem como na fertilidade dos ovos do peixe naleh, em que o dimetilsulfóxido apresentou o melhor resultado com os valores de 47,17%, 50,13% e 45,67%, respectivamente. Por outro lado, a fragmentação do DNA não ocorreu nas amostras de esperma fresco e criopreservado, indicando o efeito protetor dos crioprotetores testados. A conclusão obtida foi que o dimetilsulfóxido e 15% de gema de ovo foram o melhor crioprotetor para os espermatozoides do peixe naleh.(AU)

Cryopreservation of Trichomonas gallinae trophozoites / Criopreservação de trofozoítos de Trichomonas gallinae

This study aimed at evaluating the viability of T. gallinae isolates with the use of cryoprotectants – DMSO, ethylene glycol (EG), glycerol (GL) and propylene glycol (PG) in a freezer, in nitrogen and in an ultrafreezer, 120 days. Cryopreservation with GL, the freezing process was only viable in an ultrafreezer (20%). The use of DMSO led to viable trophozoites (40%) when freezing took place in an ultrafreezer and in nitrogen. Freezing was viable when both cryoprotectants EG (90%) and PG (80%) were used in an ultrafreezer and in nitrogen.

Este estudo teve como objetivo avaliar a viabilidade de isolados de T. gallinae com o uso de crioprotetores - DMSO, etileno glicol (EG), glicerol (GL) e propileno glicol (PG) em freezer, nitrogênio e ultrafreezer, por 120 dias. Criopreservação com GL, o processo de congelamento só foi viável em um ultrafreezer (20%). O uso de DMSO levou a trofozoítos viáveis (40%) quando o congelamento ocorreu em um ultrafreezer e em nitrogênio. O congelamento foi viável quando ambos os crioprotetores, EG (90%) e PG (80%), foram utilizados em um ultrafreezer e em nitrogênio.

Cryopreservation of Trichomonas gallinae trophozoites

Este estudo teve como objetivo avaliar a viabilidade de isolados de T. gallinae com o uso de crioprotetores - DMSO, etileno glicol (EG), glicerol (GL) e propileno glicol (PG) em freezer, nitrogênio e ultrafreezer, por 120 dias. Criopreservação com GL, o processo de congelamento só foi viável em um ultrafreezer (20%). O uso de DMSO levou a trofozoítos viáveis (40%) quando o congelamento ocorreu em um ultrafreezer e em nitrogênio. O congelamento foi viável quando ambos os crioprotetores, EG (90%) e PG (80%), foram utilizados em um ultrafreezer e em nitrogênio.(AU)

This study aimed at evaluating the viability of T. gallinae isolates with the use of cryoprotectants DMSO, ethylene glycol (EG), glycerol (GL) and propylene glycol (PG) in a freezer , in nitrogen and in an ultrafreezer, 120 days. Cryopreservation with GL, the freezing process was only viable in an ultrafreezer (20%). The use of DMSO led to viable trophozoites (40%) when freezing took place in an ultrafreezer and in nitrogen. Freezing was viable when both cryoprotectants EG (90%) and PG (80%) were used in an ultrafreezer and in nitrogen.(AU)

Cryopreservation of Trichomonas gallinae trophozoites / Criopreservação de trofozoítos de Trichomonas gallinae

This study aimed at evaluating the viability of T. gallinae isolates with the use of cryoprotectants DMSO, ethylene glycol (EG), glycerol (GL) and propylene glycol (PG) in a freezer, in nitrogen and in an ultrafreezer, 120 days. Cryopreservation with GL, the freezing process was only viable in an ultrafreezer (20%). The use of DMSO led to viable trophozoites (40%) when freezing took place in an ultrafreezer and in nitrogen. Freezing was viable when both cryoprotectants EG (90%) and PG (80%) were used in an ultrafreezer and in nitrogen.(AU)

Este estudo teve como objetivo avaliar a viabilidade de isolados de T. gallinae com o uso de crioprotetores - DMSO, etileno glicol (EG), glicerol (GL) e propileno glicol (PG) em freezer, nitrogênio e ultrafreezer, por 120 dias. Criopreservação com GL, o processo de congelamento só foi viável em um ultrafreezer (20%). O uso de DMSO levou a trofozoítos viáveis (40%) quando o congelamento ocorreu em um ultrafreezer e em nitrogênio. O congelamento foi viável quando ambos os crioprotetores, EG (90%) e PG (80%), foram utilizados em um ultrafreezer e em nitrogênio.(AU)

Cryopreservation of Trichomonas gallinae trophozoites

Este estudo teve como objetivo avaliar a viabilidade de isolados de T. gallinae com o uso de crioprotetores - DMSO, etileno glicol (EG), glicerol (GL) e propileno glicol (PG) em freezer, nitrogênio e ultrafreezer, por 120 dias. Criopreservação com GL, o processo de congelamento só foi viável em um ultrafreezer (20%). O uso de DMSO levou a trofozoítos viáveis (40%) quando o congelamento ocorreu em um ultrafreezer e em nitrogênio. O congelamento foi viável quando ambos os crioprotetores, EG (90%) e PG (80%), foram utilizados em um ultrafreezer e em nitrogênio.

This study aimed at evaluating the viability of T. gallinae isolates with the use of cryoprotectants – DMSO, ethylene glycol (EG), glycerol (GL) and propylene glycol (PG) in a freezer , in nitrogen and in an ultrafreezer, 120 days. Cryopreservation with GL, the freezing process was only viable in an ultrafreezer (20%). The use of DMSO led to viable trophozoites (40%) when freezing took place in an ultrafreezer and in nitrogen. Freezing was viable when both cryoprotectants EG (90%) and PG (80%) were used in an ultrafreezer and in nitrogen.

Efeitos da vitrificação na preservação morfológica do tecido testicular de gato doméstico / Vitrification effects during morphological preservation of testicular tissue from domestic cat

This study aimed to evaluate the effect of domestic cats (Felis Catus) testicular parenchyma vitrification by Ovarian Tissue Cryosystem and conventional straw. Three (n=3) adult cats were submitted to routine orchiectomy. For the vitrification, the samples were exposed to equilibrium solution composed by RPMI, containing 20% of ethylene glycol (EG) and 0,1M of sucrose at 20 °C, for 3 minutes. Subsequently, exposed to vitrification solution, containing RPMI added to 40% of EG and 0,1 M of sucrose at 20 °C, for 2 minutes. After devitrification, 10 seminiferous tubules of each treatment were analyzed by histology assay. The viability of spermatic and Sertolicells were analysed with light histology, as well as the seminiferous tubules morphometry. The vitrified groups were inferior to the control group in the morphologically integral cells analysis. However, the OTC was superior to straw in terms of morphological preservation of the germinative cells. Nevertheless, in morphometric analysis there was no statistical difference between the treatments (control, OTC and straw). Therefore, the vitrification in OTC method showed better results than vitrification in straw based on histological evaluation of germ and Sertoli cells of domestic cats.

Efeitos da vitrificação na preservação morfológica do tecido testicular de gato doméstico / Vitrification effects during morphological preservation of testicular tissue from domestic cat

This study aimed to evaluate the effect of domestic cats (Felis Catus) testicular parenchyma vitrification by Ovarian Tissue Cryosystem and conventional straw. Three (n=3) adult cats were submitted to routine orchiectomy. For the vitrification, the samples were exposed to equilibrium solution composed by RPMI, containing 20% of ethylene glycol (EG) and 0,1M of sucrose at 20 °C, for 3 minutes. Subsequently, exposed to vitrification solution, containing RPMI added to 40% of EG and 0,1 M of sucrose at 20 °C, for 2 minutes. After devitrification, 10 seminiferous tubules of each treatment were analyzed by histology assay. The viability of spermatic and Sertolicells were analysed with light histology, as well as the seminiferous tubules morphometry. The vitrified groups were inferior to the control group in the morphologically integral cells analysis. However, the OTC was superior to straw in terms of morphological preservation of the germinative cells. Nevertheless, in morphometric analysis there was no statistical difference between the treatments (control, OTC and straw). Therefore, the vitrification in OTC method showed better results than vitrification in straw based on histological evaluation of germ and Sertoli cells of domestic cats.(AU)

Taxas de maturação de oócitos congelados lentamente de caprinos / Slowly frozen caprine oocyte maturation rate

The objective was to evaluate the maturation rates of goat oocytes submitted to slow freezing in conventional medium for IVM. For this purpose, cumulus-oocyte complexes aspirated from pubic goat ovaries were classified morphologically and slowly frozen with 1.5 M ethylene glycol. After thawing the cryopreserved oocytes, those classified as viable were matured in conventional in vitro maturation medium. Evaluating the maturation rate, the percentage of matured oocytes in the group that underwent the cryopreservation process is significantly lower (17.6%) when compared to the control group (69.2%), also showing a high percentage of immature oocytes. Several oocyte injuries were found, caused by the studied cryopreservation method, interfering with their oocyte competence. Even with nuclear maturation rates, observed through the extrusion of the first polar corpuscle, the morphologies were altered in most oocytes, and further studies using new techniques and / or other cryoprotectants are necessary.

Ultraestrutura e criopreservação de sêmen de jundiá amazônico (Leiarius marmoratus) em cativeiro / Ultrastructure and sperm cryopreservation of the amazon catfish (Leiarius marmoratus) in captivity

Os objetivos do presente estudo foram analisar a ultraestrutura do espermatozoide do jundiá amazônico e avaliar a sua criopreservação com três agentes crioprotetores (metanol 10%, DMSO 10% e etilenoglicol 10%) e duas soluções ativadoras (NaCl 0,29% e NaHCO3 1%). Como diluente, foi utilizada uma solução de glicose a 5%, sendo o sêmen envasado em palhetas de 0,25mL e congelado em vapor de nitrogênio (botijão dry shipper). No sêmen fresco, o espermatozoide apresentou comprimento de 25,46±2,54µm, cabeça esférica (1,51±0,18µm), ausência de acrossoma, peça intermediária com formato cônico (0,93±0,17µm), ligeiramente assimétrica, com presença de vesículas, e flagelo único (21,48±2,45µm). O sêmen descongelado apresentou valores mais altos (P<0,05) para duração, vigor e taxa de motilidade espermática com os crioprotetores metanol 10% e DMSO 10%. A duração da motilidade espermática foi maior (P<0,05) com o ativador NaHCO3 1% (21-96 s). O sêmen de Leiarius marmoratus criopreservado com DMSO e metanol apresentou, respectivamente, 7,32±4,21% e 8,94±6,69% de taxa de motilidade. No entanto, os resultados não foram satisfatórios para estabelecer um protocolo para a espécie.(AU)

The aims of this study were to describe the spermatozoon ultrastructure and to evaluate the sperm cryopreservation of the amazon catfish with three cryoprotectant agents (10% methanol, 10% DMSO, and 10% ethylene glycol) and two activator agents (0.29% NaCl and 1% NaHCO3). Glucose 5% extender was used as a diluent solution and sperm loaded in 0.25 straws was frozen in nitrogen vapor (dry shipper). Fresh spermatozoon was 25.46±2.54µm long, the head was spherical (1.51±0.18µm) with no acrosome, the midpiece was cone shaped (0.93±0.17µm) with presence of vesicles, slightly asymmetric, and the flagellum was single (21.48±2.45µm). Post-thawed semen presented higher values (P< 0.05) for duration, vigor and sperm motility rate with cryoprotectants 10% methanol and 10% DMSO. The duration of sperm motility was longer (P< 0,05) when triggered in 1% NaHCO3 (96-21 s). Leiarius marmoratus semen cryopreserved with DMSO and methanol, presented respectively 7.32±4.21% and 8.94±6.69% of motility. However, the results were not satisfactory to establish a protocol for the specie.(AU)

Ultraestrutura e criopreservação de sêmen de jundiá amazônico (Leiarius marmoratus) em cativeiro / Ultrastructure and sperm cryopreservation of the amazon catfish (Leiarius marmoratus) in captivity

Os objetivos do presente estudo foram analisar a ultraestrutura do espermatozoide do jundiá amazônico e avaliar a sua criopreservação com três agentes crioprotetores (metanol 10%, DMSO 10% e etilenoglicol 10%) e duas soluções ativadoras (NaCl 0,29% e NaHCO3 1%). Como diluente, foi utilizada uma solução de glicose a 5%, sendo o sêmen envasado em palhetas de 0,25mL e congelado em vapor de nitrogênio (botijão dry shipper). No sêmen fresco, o espermatozoide apresentou comprimento de 25,46±2,54µm, cabeça esférica (1,51±0,18µm), ausência de acrossoma, peça intermediária com formato cônico (0,93±0,17µm), ligeiramente assimétrica, com presença de vesículas, e flagelo único (21,48±2,45µm). O sêmen descongelado apresentou valores mais altos (P<0,05) para duração, vigor e taxa de motilidade espermática com os crioprotetores metanol 10% e DMSO 10%. A duração da motilidade espermática foi maior (P<0,05) com o ativador NaHCO3 1% (21-96 s). O sêmen de Leiarius marmoratus criopreservado com DMSO e metanol apresentou, respectivamente, 7,32±4,21% e 8,94±6,69% de taxa de motilidade. No entanto, os resultados não foram satisfatórios para estabelecer um protocolo para a espécie.(AU)

The aims of this study were to describe the spermatozoon ultrastructure and to evaluate the sperm cryopreservation of the amazon catfish with three cryoprotectant agents (10% methanol, 10% DMSO, and 10% ethylene glycol) and two activator agents (0.29% NaCl and 1% NaHCO3). Glucose 5% extender was used as a diluent solution and sperm loaded in 0.25 straws was frozen in nitrogen vapor (dry shipper). Fresh spermatozoon was 25.46±2.54µm long, the head was spherical (1.51±0.18µm) with no acrosome, the midpiece was cone shaped (0.93±0.17µm) with presence of vesicles, slightly asymmetric, and the flagellum was single (21.48±2.45µm). Post-thawed semen presented higher values (P< 0.05) for duration, vigor and sperm motility rate with cryoprotectants 10% methanol and 10% DMSO. The duration of sperm motility was longer (P< 0,05) when triggered in 1% NaHCO3 (96-21 s). Leiarius marmoratus semen cryopreserved with DMSO and methanol, presented respectively 7.32±4.21% and 8.94±6.69% of motility. However, the results were not satisfactory to establish a protocol for the specie.(AU)

Taxas de maturação de oócitos congelados lentamente de caprinos / Slowly frozen caprine oocyte maturation rate

The objective was to evaluate the maturation rates of goat oocytes submitted to slow freezing in conventional medium for IVM. For this purpose, cumulus-oocyte complexes aspirated from pubic goat ovaries were classified morphologically and slowly frozen with 1.5 M ethylene glycol. After thawing the cryopreserved oocytes, those classified as viable were matured in conventional in vitro maturation medium. Evaluating the maturation rate, the percentage of matured oocytes in the group that underwent the cryopreservation process is significantly lower (17.6%) when compared to the control group (69.2%), also showing a high percentage of immature oocytes. Several oocyte injuries were found, caused by the studied cryopreservation method, interfering with their oocyte competence. Even with nuclear maturation rates, observed through the extrusion of the first polar corpuscle, the morphologies were altered in most oocytes, and further studies using new techniques and / or other cryoprotectants are necessary.(AU)

Efeito de diferentes osmolaridades do diluidor Tris-citrato na congelação com etilenoglicol ou glicerol de espermatozoides epididimários de gato doméstico (Felis catus) / Effect of different osmolarities of Tris-citrate extender on freezing epididymal sperm from domestic cat (Felis catus)with ethyleneglycol or glycerol

Objetivou-se com o estudo investigar o efeito de diferentes osmolaridades do diluidor Triscitrato em espermatozóides epididimários de gatos domésticos (Feliscatus) e a congelação com glicerol ou etilenoglicol. Foram realizados dois experimentos, com 10 gatos. No Experimento 1, avaliou-se a manutenção dos parâmetros espermáticos em diluidor tris-citrato com osmolaridades 275, 325, 375, 425, 475 e 525mOsm, nos tempos (T0= 0, T1= 30 e T2= 60 min). No Experimento 2 a congelação foi realizada utilizando as osmolaridades 325 e 375 do glicerol a 4% ou etilenoglicol a 3%, 6%. Dentre as osmolaridades, quanto à motilidade a 325 mOsm não diferiu estatisticamente com o fluido epidídimal (controle) nos três tempos e a 375 mOsm no T0 e T1, e ambas não apresentaram diferenças estatísticas entre si e entre os tempos em todos os parâmetros espermáticos. O uso de 4% de glicerol em diluidor com 375 mOsm foi superior, apresentando motilidade de 25% ± 6, vigor 4, integridade de membrana plasmática de 48% ± 9, sem diferenças estatísticas com o resfriamento e na morfologia não foram encontradas diferenças estatísticas entre as duas osmolaridades. Portanto, o Tris-citrato com 325 e 375 mOsm entre as osmolaridades testadas e pós congelação com 375 mOsm e glicerol 4% manteve os parâmetros espermáticos.(AU)

The aim of this study was to investigate the effect of different osmotic potentials of Tris-citrate extender in epididymal spermatozoa of domestic cats (Felis catus), frozen with glycerol or ethyleneglycol. Two experiments were carried out, with ten cats. In the first experiment, the influence of extender with the osmolarityof 275, 325, 375, 425, 475 and 525 mOsm on sperm parameters were evaluated. In the second experiment, slow freezing was performed using glycerol at 4% or ethyleneglycolat 3% and 6% added to extender with 325and 375 mOsm. Among the osmolarities, the motilityat 325 mOsm did not differ statistically with epididymal fluid (control) at all times evaluated and at 375 mOsmat T0 and T1, and both showed no statistical differences between each other and between the times in all sperm parameters. Glycerol 4% added to extender with 475 mOsm was superior, presenting motilityof 25% ± 6, vigor 4, plasma membrane integrity of 48% ± 9, without statistical differences with cooling and in morphology, no statistical differences were found between the two osmolarities. Therefore, 325 and 375 mOsm Triscitrate between the osmolarities tested and after freezing with 375 mOsm and 4%, glycerol maintained the sperm parameters.(AU)

Efeito de diferentes osmolaridades do diluidor Tris-citrato na congelação com etilenoglicol ou glicerol de espermatozoides epididimários de gato doméstico (Felis catus) / Effect of different osmolarities of Tris-citrate extender on freezing epididymal sperm from domestic cat (Felis catus)with ethyleneglycol or glycerol

Objetivou-se com o estudo investigar o efeito de diferentes osmolaridades do diluidor Triscitrato em espermatozóides epididimários de gatos domésticos (Feliscatus) e a congelação com glicerol ou etilenoglicol. Foram realizados dois experimentos, com 10 gatos. No Experimento 1, avaliou-se a manutenção dos parâmetros espermáticos em diluidor tris-citrato com osmolaridades 275, 325, 375, 425, 475 e 525mOsm, nos tempos (T0= 0, T1= 30 e T2= 60 min). No Experimento 2 a congelação foi realizada utilizando as osmolaridades 325 e 375 do glicerol a 4% ou etilenoglicol a 3%, 6%. Dentre as osmolaridades, quanto à motilidade a 325 mOsm não diferiu estatisticamente com o fluido epidídimal (controle) nos três tempos e a 375 mOsm no T0 e T1, e ambas não apresentaram diferenças estatísticas entre si e entre os tempos em todos os parâmetros espermáticos. O uso de 4% de glicerol em diluidor com 375 mOsm foi superior, apresentando motilidade de 25% ± 6, vigor 4, integridade de membrana plasmática de 48% ± 9, sem diferenças estatísticas com o resfriamento e na morfologia não foram encontradas diferenças estatísticas entre as duas osmolaridades. Portanto, o Tris-citrato com 325 e 375 mOsm entre as osmolaridades testadas e pós congelação com 375 mOsm e glicerol 4% manteve os parâmetros espermáticos.

The aim of this study was to investigate the effect of different osmotic potentials of Tris-citrate extender in epididymal spermatozoa of domestic cats (Felis catus), frozen with glycerol or ethyleneglycol. Two experiments were carried out, with ten cats. In the first experiment, the influence of extender with the osmolarityof 275, 325, 375, 425, 475 and 525 mOsm on sperm parameters were evaluated. In the second experiment, slow freezing was performed using glycerol at 4% or ethyleneglycolat 3% and 6% added to extender with 325and 375 mOsm. Among the osmolarities, the motilityat 325 mOsm did not differ statistically with epididymal fluid (control) at all times evaluated and at 375 mOsmat T0 and T1, and both showed no statistical differences between each other and between the times in all sperm parameters. Glycerol 4% added to extender with 475 mOsm was superior, presenting motilityof 25% ± 6, vigor 4, plasma membrane integrity of 48% ± 9, without statistical differences with cooling and in morphology, no statistical differences were found between the two osmolarities. Therefore, 325 and 375 mOsm Triscitrate between the osmolarities tested and after freezing with 375 mOsm and 4%, glycerol maintained the sperm parameters.

Vitrification of in vitro produced porcine blastocysts: influence of cryoprotectants toxicity and embryo age

Background: Porcine embryos are sensible to all assisted reproduction manipulations, especially the ones that involvecryopreservation. Despite the high cryoprotectant concentrations routinely applied, vitrification is the most effective technique to date. These substances toxicity can also play a negative role in embryo viability. During in vitro porcine embryoproduction, the speed of development is often unevenly distributed. It is possible that their development speed, affectsembryo tolerance to cryoprotectants. This study aimed to evaluate the toxicity of porcine embryos of days 5 or 6 of cultureto cryoprotectant agents; as well as to assess embryo survival to vitrification.Materials, Methods & Results: Parthenogenetic porcine blastocysts and expanded blastocysts of days 5 and 6 of culturewere exposed to toxicity tests (experiments 1 and 2) and vitrification (experiment 3) using different protocols. In the firstexperiment, three different cryoprotectants were used (Dimethyl sulfoxide - DMSO, Ethylene glycol - EG, and Sucrose- SUC), combined in three different associations (G1: 15% EG + 15% DMSO with 0.5 M SUC; G2: 16% EG + 16%DMSO with 0.4 M SUC; G3: 18% EG + 18% DMSO with 0.5 M SUC). In the fresh Control, embryos of day 6 are moresensible than the ones of day 5, whom showed a lower hatching rate (39.7 vs. 60.8%). After the toxicity (Experiment 1)test, the G1 showed better expansion rates in day 6 (50.0 vs 31.0 and 3.6% for G2 and G3) and higher hatching of day 6compared to G2 and G3 (23.2, vs. 8.6 and 0.0% for G2 and G3). The fresh non hatched embryos at day 8, derived at day6, had a lower percentage of cells with cleaved caspase-3 (20.2%) compared with the G1 (30.5%), G2 (31.4%) and G3(30.5%). The hatched embryos of day 5 from G2 had lower total cell number (TCN) compared with the day 6 hatchedembryos, whereas in G1 the TCN was not affected. The second experiment compared EG combined to one of these threeextracellular...

Vitrification of in vitro produced porcine blastocysts: influence of cryoprotectants toxicity and embryo age

Background: Porcine embryos are sensible to all assisted reproduction manipulations, especially the ones that involvecryopreservation. Despite the high cryoprotectant concentrations routinely applied, vitrification is the most effective technique to date. These substances toxicity can also play a negative role in embryo viability. During in vitro porcine embryoproduction, the speed of development is often unevenly distributed. It is possible that their development speed, affectsembryo tolerance to cryoprotectants. This study aimed to evaluate the toxicity of porcine embryos of days 5 or 6 of cultureto cryoprotectant agents; as well as to assess embryo survival to vitrification.Materials, Methods & Results: Parthenogenetic porcine blastocysts and expanded blastocysts of days 5 and 6 of culturewere exposed to toxicity tests (experiments 1 and 2) and vitrification (experiment 3) using different protocols. In the firstexperiment, three different cryoprotectants were used (Dimethyl sulfoxide - DMSO, Ethylene glycol - EG, and Sucrose- SUC), combined in three different associations (G1: 15% EG + 15% DMSO with 0.5 M SUC; G2: 16% EG + 16%DMSO with 0.4 M SUC; G3: 18% EG + 18% DMSO with 0.5 M SUC). In the fresh Control, embryos of day 6 are moresensible than the ones of day 5, whom showed a lower hatching rate (39.7 vs. 60.8%). After the toxicity (Experiment 1)test, the G1 showed better expansion rates in day 6 (50.0 vs 31.0 and 3.6% for G2 and G3) and higher hatching of day 6compared to G2 and G3 (23.2, vs. 8.6 and 0.0% for G2 and G3). The fresh non hatched embryos at day 8, derived at day6, had a lower percentage of cells with cleaved caspase-3 (20.2%) compared with the G1 (30.5%), G2 (31.4%) and G3(30.5%). The hatched embryos of day 5 from G2 had lower total cell number (TCN) compared with the day 6 hatchedembryos, whereas in G1 the TCN was not affected. The second experiment compared EG combined to one of these threeextracellular...(AU)