Effects of time-dependent degradation and freezing on biomechanical properties of healthy bovine jejunum / Efeitos da degradação dependente do tempo e do congelamento sobre as propriedades biomecânicas do jejuno bovino saudável

Degradation of bovine small intestine and respective effects on biomechanics have not been described to date. Biomechanical testing of intestinal tissues is often carried out within a few hours of donor death and tissue deterioration is not accounted for. Freezing is efficient for the preservation of several tissues; however, it may cause cellular damage. This study investigated the morphologic and biomechanical changes of bovine jejunum at different postmortem moments. Effects of freezing and thawing on morphology and biomechanical behavior were also examined. Macroscopic changes were first noted within eight hours of death. At this time, histologic changes also started to set in, and biomechanical tests revealed lower bursting pressure (203.10±46.14mmHg). At 12 hours, tissue rearrangement was noted, and bursting pressure increased (238.43±31.04mmHg). A second drop in pressure was detected at 18 hours (235.20±38.21mmHg), followed by a progressive drop until the end of the experimental period. Histologic changes revealed progressive deterioration. Mechanical resistance did not differ between thawed and fresh specimens. It was concluded that bovine jejunal specimens retain biomechanical resistance up to 6 hours after death. Freezing and thawing did not affect the mechanical resistance of the intestinal wall in this experimental model.

A degradação do intestino delgado de bovinos e seu efeito biomecânico não são descritos na literatura. Trabalhos que envolvem a biomecânica intestinal realizam os ensaios em poucas horas após o óbito dos doadores, sem avaliação de seu estado de deterioração. O congelamento é eficiente na conservação de vários tecidos, porém pode provocar danos em nível celular. Este estudo teve por objetivo avaliar a degradação morfológica e biomecânica do jejuno de bovinos em diferentes tempos post mortem. O efeito do congelamento e do descongelamento sobre essas características também foi avaliado. As primeiras alterações macroscópicas foram observadas oito horas após o óbito. No mesmo momento, se iniciaram as alterações histológicas e a menor pressão suportada ao teste biomecânico (203,10±46,14mmHg). A partir de 12 horas, o tecido sofreu um rearranjo, suportando maior pressão (238,43±31,04mmHg). Uma segunda queda foi detectada após 18 horas (235,20±38,21mmHg), seguida de redução progressiva até o término do experimento. As lesões histológicas foram progressivas e graduais. As amostras testadas após descongelamento não apresentaram diferença estatística quanto à resistência mecânica em comparação com os espécimes frescos. Concluiu-se que as amostras mantêm sua resistência biomecânica até 6 horas após o óbito. Também se mostrou que o congelamento e o descongelamento, nas condições testadas, não alteraram a resistência mecânica da parede intestinal.

Critérios e metodologias para produzir sêmen de caprinos e ovinos em nível de fazenda / The criteria and methodologies used to produce ram and goat semen on the farm

A criopreservação de sêmen é uma importante estratégia para o armazenamento de material genético de reprodutores ovinos e caprinos, considerados superiores do ponto de vista fenotípico, mas especialmente, pelos seus méritos genéticos. Hoje no Brasil apenas uma central regulamentada pelo MAPA está ema atividade, o que significa que existe pouca dose de sêmen comercial à disposição para programas de melhoramento genético. Programas de banco de sêmen podem ser realizado nas fazendas, entretanto, tem seu uso limitado a propriedade onde está o reprodutor. Neste caso, apesar de maiores limitações de estrutura e equipamentos para a execução do serviço, o domínio das variações da técnica de congelação de sêmen e o cuidado com as questões sanitárias dos reprodutores possibilitará ao Médico-Veterinário a execução do serviço com níveis adequados de segurança e qualidade das doses de sêmen produzidas. O objetivo deste estudo é apresentar e discutir critérios e metodologias para congelação de sêmen de pequenos ruminantes em diferentes condições, fruto de diversas experiências e trabalhos desenvolvidos por nosso grupo e outros, nos últimos vinte anos, e que tem possibilitado resultados satisfatórios.(AU

Sperm cryopreservation is an important strategy for the storage of genetic material from ovine and goat breeders, considered superior from the phenotypic point of view, but especially for their genetic merits. Today in Brazil only one center regulated by the Ministry of Agriculture, Livestock and Supply is in operation, which means that there is little dose of commercial semen available for genetic improvement programs. Semen bank programs can be carried out on farms; however, their use is limited to the property where the breeder is located. In this case, despite greater limitations of structure and equipment for the execution of the service, mastering variations in the semen freezing technique and taking care of the health issues of the breeders will enable the Veterinarian to perform the service with adequate levels of safety and quality of semen doses produced. This study aims to present and discuss criteria and methodologies for freezing semen from small ruminants under different conditions, the result of several experiences and works carried out by our group and others, in the last twenty years, which have enabled satisfactory results.(AU))

Effect of heat stress and solar radiation on dry matter intake, biochemical indicators, production, and quality of Holstein and Jersey cows' milk / Efeito do estresse calórico e da insolação sobre consumo, indicadores bioquímicos, produção e qualidade do leite de vacas Holandês e Jersey

The objective of this work was to compare the dry matter intake, milk yield and quality, physiological and biochemical parameters in Holstein (n=10) and Jersey (n=10) cows under heat stress and insolation, in two treatments: CL - cooling by ventilation and sprinkling and HS - heat stress and insolation. Data were submitted to ANOVA. There was an interaction between treatment and breed and day effect for dry matter intake. For consumption in % of body weight, CL and Jersey cows consumed more. CL cows produced more milk and 3.5% fat-corrected milk. Feed efficiency was similar between treatments and breeds. Fat, lactose, total solids, and somatic cell score did not differ. The concentration of milk urea nitrogen was higher for CL cows. Milk from Holstein cows had greater stability to alcohol, and from HT cows had a greater freezing point of milk. HT cows had higher respiratory rates in the morning and surface temperatures in the afternoon. There were no differences in beta-hydroxybutyrate and glucose concentrations. Heat stress, with insulation, reduces intake, especially in Holstein cows, as well as milk production and increases the freezing point of milk, respiratory rate, and surface temperature.

O objetivo deste trabalho foi comparar o consumo de matéria seca, a produção e a qualidade do leite, os parâmetros fisiológicos e bioquímicos em vacas das raças Holandesa (n=10) e Jersey (n=10) sob estresse calórico e insolação, em dois tratamentos: CL - resfriamento por ventilação e aspersão; HS - estresse térmico e insolação. Os dados foram submetidos à análise de variância. Houve interação entre tratamento e raça e efeito de dia para consumo de matéria seca. Para consumo em % de peso vivo, vacas CL e Jersey consumiram mais. Vacas CL produziram mais leite e leite corrigido a 3,5% de gordura. A eficiência alimentar foi similar entre tratamentos e raças. Teores de gordura, lactose, sólidos totais e escore de células somáticas não diferiram. A concentração de nitrogênio ureico do leite foi maior para vacas CL. O leite das vacas Holandesas apresentou maior estabilidade ao álcool, e de vacas HT maior crioscopia. Vacas HT apresentaram maior frequência respiratória de manhã e temperatura superficial à tarde. Não houve diferenças para concentração de beta-hidroxibutirato e glicose. O estresse calórico, com insolação, reduz o consumo, especialmente em vacas Holandesas, bem como a produção de leite, com aumento da crioscopia, elevando a frequência respiratória e a temperatura superficial.

Novel technologies combined with osmotic dehydration for application in the conservation of fruits: an overview / Novas tecnologias combinadas com desidratação osmótica para aplicação na conservação de frutas: uma revisão

ABSTRACT: Osmotic dehydration (OD) is a technique used for the partial removal of water from foodstuff, including fruit and vegetables, with the aim of producing a desiccated product. The process involves placing the material in a hypertonic solution for several hours and allowing water to move from the cell compartment into the solution by osmosis. OD is influenced by various factors such as the concentration and composition of the osmotic solution, the solution temperature, the type of agitation and the time of exposure, as well as the size, shape and compactness of the food material. The main advantages of OD over conventional drying processes are the superior quality of the dried products and the minimization of shrinkage. In recent years, research effort has focused on the combination of OD with other technologies, such as ultrasound, cryogenic freezing with liquid nitrogen, pulsed electric field, gamma radiation and high hydrostatic pressure. The application of these methods prior to or concomitant with OD accelerates mass transfer and reduces the drying rate of fruit and vegetables by increasing the permeability of cell membranes. In this manner, combined processes tend to be more efficient and economical in comparison with conventional OD because they reduce operating times and; consequently, energy consumption. In addition, the dried products generated by such coupled processes typically exhibit improved nutritional and physicochemical characteristics. This review summarizes the basic principles and applications of OD in combination with other methods, with particular emphasis on the production of dried fruits.

RESUMO: A desidratação osmótica (DO) é uma técnica utilizada para remover parcialmente a água dos alimentos, incluindo frutas e vegetais, com vistas a produção de alimentos secos. O processo consiste em colocar o material em uma solução hipertônica por várias horas e deixar a água passar do compartimento celular para a solução por osmose. A DO é influenciada por vários fatores como a concentração e composição da solução osmótica, a temperatura da solução, o tipo de agitação e o tempo de exposição, assim como o tamanho, forma e compactação do material alimentar. As principais vantagens da DO em relação aos processos de secagem convencionais são que ela dá origem a produtos secos de qualidade superior e minimiza o encolhimento. Nos últimos anos, tem-se investigado a combinação da DO com outras tecnologias, tais como ultrassom, congelamento criogênico com nitrogênio líquido, campo elétrico pulsado, radiação gama e alta pressão hidrostática. A aplicação desses métodos antes ou simultaneamente com a DO acelera a transferência de massa e reduz a taxa de secagem de frutas e vegetais através do aumento da permeabilidade das membranas celulares. Assim, os processos combinados tendem a ser mais eficientes e econômicos do que a DO convencional, pois reduzem o tempo de operação e, consequentemente, o consumo de energia. Adicionalmente, os produtos desidratados gerados através de processos associados geralmente apresentam melhores características nutricionais e físico-químicas. Esta revisão sumariza os princípios básicos e aplicações da DO em combinação com outros métodos, com ênfase especial dada à produção de frutas secas.

Kinetic vitrification: concepts and perspectives in animal sperm cryopreservation

Sperm cryopreservation is an important tool for genetic diversity management programs and the conservation of endangered breeds and species. The most widely used method of sperm conservation is slow freezing, however, during the process, sperm cells suffer from cryoinjury, which reduces their viability and fertility rates. One of the alternatives to slow freezing is vitrification, that consist on rapid freezing, in which viable cells undergo glass-like solidification. This technology requires large concentrations of permeable cryoprotectants (P- CPA's) which increase the viscosity of the medium to prevent intracellular ice formation during cooling and warming, obtaining successful results in vitrification of oocytes and embryos. Unfortunately, this technology failed when applied to vitrification of sperm due to its higher sensitivity to increasing concentrations of P-CPAs. Alternatively, a technique termed 'kinetic sperm vitrification' has been used and consists in a technique of permeant cryoprotectant-free cryopreservation by direct plunging of a sperm suspension into liquid nitrogen. Some of the advantages of kinetic vitrification are the speed of execution and no rate-controlled equipment required. This technique has been used successfully and with better results for motility in human (50-70% motility recovery), dog (42%), fish (82%) and donkey (21.7%). However, more studies are required to improve sperm viability after devitrification, especially when it comes to motility recovery. The objective of this review is to present the principles of kinetic vitrification, the main findings in the literature, and the perspectives for the utilization of this technique as a cryopreservation method.(AU)

Efecto de la morfología espermática y la concentración de colesterol y triglicéridos del plasma seminal sobre la supervivencia espermática del semen congelado en felinos / Effect of sperm morphology and seminal plasma cholesterol and triacylglycerides concentrations on semen freezability in felines

Se ha documentado la gran variedad y el alto porcentaje de morfoanomalías espermáticas presentes en gatos fértiles. Estudios recientes han demostrado que el semen de buena calidad tiene una alta concentración de colesterol (CHOL) y triglicéridos (TAG) en gatos. Sin embargo, en el gato doméstico hay escasa información sobre el efecto de la composición bioquímica del plasma seminal y la morfología espermática sobre la supervivencia espermática luego de la congelación y descongelación del semen. Estudios recientes sugieren que concentraciones plasmáticas seminales más altas de CHOL y TAG podrían mejorar la capacidad de congelación del semen. Sin embargo, no hay evidencia de que la morfología de los espermatozoides afecte la resistencia de los espermatozoides a la criopreservación.(AU)

The great variety and high percentage of sperm morpho-anomalies present in fertile cats have been documented. Recent studies have shown that good-quality semen has high cholesterol (CHOL) and triglycerides (TAG) in cats. However, in the domestic cat, there is little information on the effect of seminal plasma biochemical composition and sperm morphology on sperm survival after semen frozen-thawed. Recent studies suggest that higher seminal plasma concentrations of CHOL and TAG could improve semen freezing. However, no evidence exists that sperm morphology affects sperm resistance to cryopreservation.(AU)

Seleção de reprodutores caprinos e ovinos para produção de sêmen / Selection of caprine and ovine sires for semen production

A presente revisão propôs analisar quais características são desejáveis para seleção de reprodutores caprinos e ovinos, visando à produção de sêmen. A escolha dos reprodutores deve ser feita de acordo com os padrões exigidos da raça, além da higidez reprodutiva dos animais. Biotecnologias reprodutivas oferecem oportunidades consideráveis para a produção animal, como a inseminação artificial, transferência de embriões e congelamento de sêmen. Análises da produção de espermatozoides são de grande importância, pois está diretamente relacionada com a atividade sexual. A genética do criatório é um fator crítico que influencia o resultado final e a saúde animal. A utilização de marcadores moleculares é uma ferramenta que temos para selecionar características desejáveis em uma prole, através da identificação de biomarcadores. Técnicas de genética molecular potencializam o efeito de biotécnicas reprodutivas e consequentemente a lucratividade da pecuária intensiva.(AU)

The present review proposed to analyze which characteristics are desirable for the selection of goat and sheep breeders, aiming at the production of semen. The choice of breeders must be made in accordance with the standards required for the breed, in addition to the reproductive health of the animals. Reproductive biotechnologies offer considerable opportunities for animal production, such as artificial insemination, embryo transfer and semen freezing. Analyzes of sperm production are of great importance, as it is directly related to sexual activity. Farm genetics is a critical factor that influences the end result and animal health. The use of molecular markers is a tool that we have to select desirable characteristics in an offspring, through the identification of biomarkers. Molecular genetic techniques enhance the effect of reproductive biotechniques and consequently the profitability of intensive livestock farming.(AU)

Criopreservação e cultivo de tecido testicular em mamíferos - estado da arte, desafios e perspectivas / Cryopreservation and culture of testicular tissue in mammals - state of the art, challenges and perspectives

O tecido testicular contém células germinativas, que possuem potencial para se desenvolver em espermatozoides viáveis por meio do cultivo in vitro ou xenotransplante, sendo uma alternativa interessante a ser utilizada na formação de biobancos. Esta revisão compila as atualizações, desafios e perspectivas relacionadas às técnicas de criopreservação e cultivo de tecido testicular como estratégia para a conservação de espécies mamíferas. O tecido testicular pode ser obtido tanto de indivíduos adultos como pré púberes, seja após orquiectomia ou até mesmo após a sua morte. O tecido fragmentado pode ser criopreservado por congelação lenta ou rápida e por métodos de vitrificação. Os crioprotetores são indispensáveis durante a criopreservação e podem variar o tipo e concentração de acordo com a espécie. Com os avanços da criopreservação deste material, espermatozoides podem ser obtidos por transplante de fragmentos testiculares ou células germinativas isoladas em camundongos imunodeficientes. No entanto, a obtenção de espermatozoides no cultivo in vitro ainda é um desafio.(AU)

The testicular tissue contains germ cells, which have the potential to develop into viable spermatozoa through in vitro culture or xenotransplantation, being an interesting alternative to be used in the formation of biobanks. This review compiles updates, challenges and perspectives related to cryopreservation techniques and testicular tissue culture as a strategy for the conservation of mammalian species. Testicular tissue can be obtained from both adult and pre-pubertal individuals, either after orchiectomy or even after their death. Fragmented tissue can be cryopreserved by slow or fast freezing and by vitrification methods. Cryoprotectants are indispensable during cryopreservation and may vary in type and concentration according to the species. With advances in cryopreservation of this material, spermatozoa can be obtained by transplanting testicular fragments or isolated germ cells into immunodeficient mice. However, obtaining spermatozoa in in vitro culture is still a challenge.(AU)

Effect of milking hygiene, herd size, water hardness and temperature-humidity index on milk quality of dairy farms

The objective of this study was to evaluate the effect of milking hygiene practices, herd size, water hardness, and temperature-humidity index (THI) on the physicochemical and microbiological characteristics of raw milk, and standard plate count (SPC) in milking machines of dairy farms in the central region of Mexico. Data were collected from fifty-three dairy farms during one year. The evaluated effects included milking hygiene conditions (good, medium, poor), herd size (1-50, 51-100, 101-150, ≥151 heads), water hardness (soft or moderately hard), and THI (comfortable or stressful). The increase in milking hygiene produced greater milk yield (MY) and energy corrected milk (ECM) but lower protein content, and decreased the individual bacterial count (IBC) and somatic cell count (SCC). The MY, ECM, protein content, IBC, and SCC were higher on bigger farms. The use of soft water reduced MY, IBC, and SCC, but improved fat, lactose, total solids (TS), and non-fat solids (NFS). Heat stress negatively affected fat, protein, TS, NFS, acidity, freezing point (FP), SCC, and methylene blue dye reduction test. Poor milking hygiene contributes to higher SPC in milking machine parts. Water hardness and THI did not affect SPC in all milking machine parts. Proper milking hygiene practices, larger herd size, softer water, lower THI, and adequate cleaning and disinfection of the milking machine parts benefits the physicochemical and microbiological quality of the milk.

Viability of Toxoplasma gondii in cattle semen cryopreserved with different concentrations of cryoprotectant / Viabilidade do Toxoplasma gondii em sêmen de bovino criopreservado em diferentes concentrações de crioprotetores

ABSTRACT: Toxoplasma gondii can be eliminated in bovine semen. Cryopreserved semen is often used due to the fact that artificial insemination in dairy and beef cattle provides benefits in terms of production. However, little is known regarding the viability and infectivity of T. gondii tachyzoites in cryopreserved bovine semen. In the present study, cattle semen negative for T. gondii were contaminated with 1 x 106 tachyzoites (RH strain) and cryopreserved with and without different cryoprotectants, such as DMSO (concentrations of 2.5%, 5.0%, 7.5%, 8.0% and 10.0%) and glycerol (2.25%, 2.5%, 3.0%, 5.0%, 7.5% and 10.0%), followed by freezing in liquid nitrogen (-196°C). After 24 hours, the samples were thawed and inoculated in 10 mice per cryoprotectant concentration. The mice were evaluated for clinical signs of toxoplasmosis (rough coat, diarrhea, hypoactivity and sudden death) as well as serum titers of IgM and IgG and the presence of tachyzoites in the peritoneal lavage. The results revealed that T. gondii remained infective in all samples. Clinical signs of toxoplasmosis were observed in the mice beginning with the 6th day post-inoculation (DPI) and 100% lethality was found between the 7th and 9th DPI. Viable tachyzoites were recovered from peritoneal exudate of dead mice (except for the control group), with higher mean of tachyzoite counts in the intraperitoneal lavage for 5% DMSO (±3.32 x 106), 8% DMSO (±3.53 x 106), 3% glycerol (±4.75 x 106), 7.5% glycerol (±6.26 x 106) and the absence of cryoprotectant (±3.11 x 106). Seroconversion occurred in the treated groups, with titers of IgG from 1:16 to 1:128 and IgM from 1:16 to 1:512. T. gondii viability and infectivity were maintained in cattle semen during 24 hours of cryopreservation at -196°C with and without cryoprotectant. However, further studies are necessary to determine whether cryopreserved semen contributes to the spread of toxoplasmosis through artificial insemination.

RESUMO: Sabe-se que Toxoplasma gondii pode ser eliminado no sêmen bovino. A inseminação artificial em bovinos leiteiros e de corte proporcionou avanços e benefícios nas produções e para isso o sêmen criopreservado é frequentemente utilizado. No entanto, pouco se sabe sobre a viabilidade e infectividade dos taquizoítos de T. gondii em sêmen bovino criopreservado. Para isso o sêmen bovino, negativo para T. gondii, foi contaminado com 1x106 taquizoítos (cepa RH), criopreservados com ou sem diferentes crioprotetores como DMSO (2.5%, 5.0%, 7.5%, 8.0% e 10.0%) e Glicerol (2.25%, 2.5%, 3.0%, 5.0%, 7.5% e 10.0%) e congelados em nitrogênio líquido (-196°C). Após 24 horas, essas amostras foram descongeladas e inoculadas em 10 camundongos por diluente de concentração de crioprotetor. Os camundongos foram avaliados quanto a sinais clínicos de toxoplasmose (pele áspera, diarreia, hipoatividade e morte súbita), títulos séricos de IgM e IgG e presença de taquizoítos no lavado peritoneal. Os resultados mostraram que T. gondii se manteve infectante em todas as amostras, inclusive naquelas sem crioprotetor. Sinais clínicos de toxoplasmose foram observados nos camundongos a partir do 6º dia pós-inoculação (DPI) e 100% de letalidade foi verificada entre o 7º ao 9º DPI. Nos camundongos mortos, exceto no grupo controle, taquizoítos viáveis foram recuperados do exsudato peritoneal, com maior média de taquizoítos quantificados na lavagem intraperitoneal para DMSO a 5% (±3.32x106), 8% (±3.53x106) e glicerol 3% (±4.75x106), 7,5% (±6.26x106) e livre de crioprotetor (±3.11x106). A soroconversão ocorreu nos grupos tratados com títulos de IgG (1:16 a 1:128) e IgM (1:16 a 1:512). A viabilidade e infectividade do T. gondii no sêmen bovino durante as 24 horas de criopreservação a -196°C foram mantidas com ou sem crioprotetor. No entanto, mais estudos são necessários para verificar se o sêmen criopreservado contribui para a disseminação da toxoplasmose na inseminação artificial.

Criopreservação do sêmen suíno: desafios a serem superados que auxiliam a identificação de espermatozoides mais férteis / Boar semen cryopreservation: challenges to be overcome that help the identification of more fertile spermatozoa

A criopreservação é o melhor método de armazenamento de sêmen por período indeterminado, conferindo alta flexibilidade de uso. Além disso, é o principal meio para formação de banco de germoplasma. No entanto, congelar sêmen suíno ainda é um desafio a ser enfrentado. Isso porquê, além das características de membrana espermática que ocasionam maiores efeitos deletérios às células durante os processos de congelação e descongelação, ainda é preciso lidar com a grande variabilidade de resultados, seja entre indivíduos, ejaculados de um mesmo indivíduo ou ainda, frações de um mesmo ejaculado. Frente a isso, diferentes estratégias têm sido investigadas a fim de obter melhores resultados e assim, aumentar a aplicabilidade do sêmen congelado suíno a nível comercial. Além disso, acreditamos que haja a possibilidade de se relacionar a capacidade de criotolerância de um determinado espermatozoide a sua maior ou menor capacidade de fertilizar o oócito.(AU)

Cryopreservation is the best method of storing semen indefinitely, providing high flexibility of use. In addition, it is the primary means for forming a germplasm bank. However, freezing boar semen is still a challenge to be faced because, in addition to the characteristics of the spermatic membrane that cause deleterious effects to the cells during the freezing and thawing processes, it is still necessary to deal with the significant variability of results, whether between individuals, ejaculates from the same individual or even fractions of an even ejaculated. Because of this, different strategies have been investigated to obtain better results and thus increase the applicability of frozen boar semen at a commercial level. Furthermore, we believe it is possible to relate the cryotolerance capacity of a given sperm to its greater or lesser capacity to fertilize the oocyte.(AU)

Sêmen refrigerado ­ estado da arte em diferentes espécies / Chilled semen ­ state of the art in different species

Na atual conjuntura da criação artificial de bovinos e bubalinos, o material genético masculino de qualidade superior de reprodutores é explorado ao máximo possível através da inseminação artificial em tempo fixo de um grande número de fêmeas com apenas um único ejaculado. Para isso, é necessário um sêmen de boa qualidade que desempenhe um papel indispensável na melhoria das taxas de fertilidade, independente de qual tipo seja utilizado (fresco, refrigerado e congelado). Porém, o processo de congelação/descongelação causa uma série de injúrias aos espermatozoides, ocasionando resultados inferiores para percentuais de viabilidade espermática, motilidade, membrana plasmática e integridade acrossomal, potencial de membrana mitocondrial, cinemática do esperma, quando comparado ao sêmen refrigerado. Assim, o objetivo desta revisão é disseminar o conhecimento sobre o uso sêmen refrigerado na preservação de germoplasma de reprodutores bovinos e bubalinos para melhorar as taxas de concepção em propriedades. Para isso, serão abordados comentários sobre o armazenamento do sêmen refrigerado, com ênfase nas diferenças entre curvas de refrigeração, suas vantagens e desvantagens relativas para procedimentos de uso na IATF, identificando o método mais indicado por diversos autores, o estado atual da biotécnica, seus méritos e possibilidades futuras.(AU)

In the current conjuncture of the artificial creation of bovines and buffaloes, the male genetic material of superior quality of sires is exploited to the maximum possible through the fixed-time artificial insemination of a large number of females with only a single ejaculate. For this, a good quality semen is needed that plays an indispensable role in improving fertility rates, regardless of which type is used (fresh, chilled and frozen). However, the freezing/thawing process causes a series of injuries to spermatozoa, causing lower results for percentages of sperm viability, motility, plasma membrane and acrosomal integrity, mitochondrial membrane potential, sperm kinematics, when compared to refrigerated semen. Thus, the objective of this review is to disseminate knowledge about the use of chilled semen in the preservation of germplasm of bovine and buffalo breeders to improve conception rates in properties. For this, comments on the storage of refrigerated semen will be addressed, with emphasis on the differences between refrigeration curves, their relative advantages and disadvantages for procedures for use in FTAI, identifying the method most indicated by several authors, the current state of biotechnics, its future merits and possibilities.(AU)

Camellia sinensis extract delays microbiological and oxidative changes in striped catfish fillets during frozen storage / Extrato de chá verde age sobre a qualidade de filés de pescado durante o armazenamento por congelamento

This study investigated the effects of tea leaf (Camellia sinensis) extract on the quality of striped catfish (Pangasianodon hypophthalmus) fillets during 18-months of frozen storage (-20 ± 2 °C). Fillet samples were submitted to the treatments Control (cold tap water), CS 7.63 (C. sinensis extract solution 7.63 µg / mL) and CS 625 (C. sinensis extract 625 µg / mL) and stored for 18 months, with collections performed at 0, 1, 3, 6, 9, 12 and 18 months. Total viable count, physicochemical parameters (water holding capacity, total volatile basic nitrogen, peroxide value, thiobarbituric acid reactive substances, moisture and pH), sensory properties and color measurement were evaluated. Results showed that fillets treated with C.a sinensis extracts slightly reduced lipid oxidation, inhibited bacterial growth and improved sensory properties compared to untreated samples, without causing significant changes in the other quality indicators. The findings indicated that the green tea leaf extract immersion treatments, contributed to the improved quality preservation of striped catfish fillets during frozen storage.

O objetivo deste estudo foi investigar os efeitos do extrato de folhas de chá verde (Camellia sinensis) na qualidade de filés de bagre listrado (Pangasianodon hypophthalmus) durante 18 meses de armazenamento congelado (-20 ± 2 ° C). O estudo incluiu três tratamentos: imersão dos filés de bagre listrado em água fria da torneira como um tratamento de controle, em solução de extrato de Camellia sinensis 7,63 µg / mL e em solução de extrato de Camellia sinensis 625 µg / mL. As amostras foram armazenadas por 18 meses e as coletas foram feitas aos zero, um, três, seis, nove, 12 e 18 meses. Os parâmetros avaliados incluíram contagem viável total, parâmetros físico-químicos (capacidade de retenção de água, nitrogênio básico volátil total, valor de peróxido, substâncias reativas ao ácido tiobarbitúrico, umidade e pH), propriedades sensoriais e medição de cor. Os resultados mostraram que os filés de bagre listrado tratados com extratos de Camellia sinensis reduziram ligeiramente a oxidação de lipídios, inibiram o crescimento de bactérias e aumentaram as propriedades sensoriais em comparação com as amostras não tratadas. Além disso, o tratamento do extrato de Camellia sinensis não afetou o pH, a umidade, a capacidade de retenção de água, o nitrogênio básico volátil total e a cor do filé durante o armazenamento congelado. Com base na contagem viável total, parâmetros físico-químicos e qualidade sensorial, pode-se concluir que filés de bagre listrados não tratados ou tratados com extratos de Camellia sinensis (7,63 e 625 µg / mL) podem ser usados ​​por até 18 meses.

Dimethyl acetamide and dimethyl sulfoxide associated at glucose and egg yolk for cryopreservation of Pseudoplatystoma corruscans semen

This study aimed to develop a protocol for the cryopreservation of Pseudoplatystoma corruscans semen. For this, mature males were hormonally induced with a single dose of carp pituitary extract (5 mg/kg body weight). Semen was collected and evaluated. Two cryoprotectants were tested to compose the diluents: dimethyl acetamide (DMA) and dimethyl sulfoxide (Me2SO), in two concentrations (8% and 10%), + 5.0% glucose + 10% egg yolk. The semen was diluted in a 1: 4 ratio (semen: extender), packed in 0.5 mL straws and frozen in a dry shipper container in liquid nitrogen vapors. After thawing, sperm kinetics, sperm morphology and DNA integrity of cryopreserved sperm were evaluated. Pseudoplatystoma corruscans males produced semen with sperm motility > 80%. After thawing, all treatments provided semen with total sperm motility > 40%, with no significant difference (P < 0.05) between them, as well as between the other sperm kinetic parameters evaluated. The treatments with DMA provided a smaller fragmentation of the DNA of the gametes. Sperm malformations were identified in both fresh and cryopreserved semen, with a slight increase in these malformations being identified in sperm from thawed P. corruscans semen samples.(AU)

Este estudo teve como objetivo desenvolver um protocolo para a criopreservação do sêmen de Pseudoplatystoma corruscans. Para tal, machos maduros foram induzidos hormonalmente com uma dose única de extrato de hipófise de carpa (5 mg/kg de peso vivo). O sêmen foi coletado e avaliado. Sendo testados para compor os diluentes, dois crioprotetores: dimetil acetamida (DMA) e dimetil sulfóxido (Me2SO), em duas concentrações (8% e 10%), + 5,0% glicose + 10% gema de ovo. O sêmen foi diluído na proporção 1: 4 (sêmen: extensor), embalado em palhetas de 0,5 mL e congelado em container dryshipper em vapores de nitrogênio líquido. Após o descongelamento, foram avaliados os aspectos cinéticos espermáticos, a morfologia espermática e a integridade do DNA dos espermatozoides criopreservados. Os machos de P. corruscans produziram sêmen com motilidade espermática > 80%. Todos os tratamentos proporcionaram após o descongelamento sêmen com motilidade espermática total > 40%, sem diferença significativa (P < 0,05) entre eles, como também entre os demais parâmetros cinéticos espermáticos avaliados. Os tratamentos com DMA proporcionaram uma menor fragmentação do DNA dos gametas. Malformações espermáticas foram identificadas, tanto no sêmen fresco, como no criopreservado, sendo identificado um aumento discreto dessas malformações nos espermatozoides das amostras de sêmen descongeladas de P. corruscans.(AU)

Cryopreservation of jaguar (Panthera onca) sperm cells using different cryoprotectants and different thawing temperatures

The cryopreservation of jaguar semen must be improved to produce high-quality biobanking doses. Until now, the rare studies of semen freezing in the species have only evaluated glycerol, always with a significant reduction in sperm quality in thawed semen. The purpose of this study was to assess the efficacy of three cryoprotectants, dimethylsulfoxide (DMSO), glycerol (GLY), and methanol (MET), in the cryopreservation of jaguar semen in an LDL-based extender, as well as the effect of thawing temperature on dosage quality. Five mature males with a history of reproduction were used. On the males, an infrared thermal image (IRT) was captured, the spicules and testes were analyzed, and the CASA system was used to evaluate the quality of fresh and thawed sperm. The superficial IRT was 4.6 ± 1.2 °C cooler than the anal sphincter, and the semen measured between 27.3 and 28.7 °C shortly after exiting the urethra. The total motility of fresh sperm was 55.3 ± 22.6%, and progressive motility was 36.3 ± 18%. The total motility of thawed sperm was 5.28 ± 2.51%, 4.49 ± %2.49, and 0.51 ± 0.62% for DMSO, GLY, and MET, respectively. DMSO and GLY performed better than MET, and there was no difference in thawing temperature (37°C 30 s vs. 50°C 12 s). All animals exhibit a considerable level of morphological changes in sperm. Low amounts of total and progressive motility were found in the thawed sperm. Males with a high level of sperm morphological changes were found to be fertile, but the lone male with normospermia was infertile. Thus, we contest the applicability of the commonly used morphological classification for bovines to felid species.(AU)

Impact of quercetin, carnosine, and ozone in the cryopreservation on Nellore (Bos indicus) semen

The objective of this study was to reduce the effects of cryoinjury caused in bovine semen by cryopreservation. Ejaculates were collected from Nellore bulls and subjected to freezing in C (control), ozone (15, 30, and 60 µg mL-1 of ozone), quercetin (25, 50, and 100 µg mL-1 of quercetin), and carnosine groups (100, 200, and 300 ng mL-1 of carnosine). Samples were evaluated post-thaw (M0) and post-rapid thermoresistance test (M30) for sperm kinetics (total motility, progressive motility, curvilinear speed, linearity and amplitude of lateral head displacement) and cell structure viability (plasma membrane integrity, acrosomal integrity, mitochondrial potential, membrane fluidity, and lipid peroxidation). There were no differences (P > 0.05) between the control, quercetin, and carnosine-treated groups for the parameters evaluated at M0 and M30. In turn, supplementation with ozone resulted in lower values for sperm kinetics (P < 0.05) and lower mitochondrial potential at M30 (P < 0.05). Quercetin and carnosine at the concentrations used did not promote significant gains in frozen semen, nor did they demonstrate cytotoxicity. We expected to obtain positive results with the use of ozone. Nonetheless, the addition was harmful to the parameters of sperm kinetics, and its effect was not observed as a possible pro-antioxidant. We believe that the fact that the gas did not harm the sperm structure opens avenues for tests with lower dosages, since, by reducing its concentration, we could minimize the damage to sperm kinetics.(AU0

Comparative study of bullfrog (Rana catebeiana) skin preservation for use as a biomaterial / Estudo comparativo de meios de conservação de pele de rã-touro (Rana catesbeiana) para utilização como biomaterial

The use of biological membranes in wound dressings has increasingly become a reality. Accordingly, an ideal means of preservation is sought that can provide tissue maintenance for long periods without interfering with its quality or clinical applicability. Therefore, the objective of the present study was to evaluate and histologically and microbiologically compare frog skins subjected to two different preservation methods. Sixteen frog skins were evaluated and, depending on the preservation method, subdivided into two groups with eight skins each, namely, the Freezing Group, in which the skins were frozen at -4º in a 20% glycerin solution; and the Glycerin Group, whose skins were kept in 98% glycerin at room temperature (average of 28 ºC and average humidity of 78%). The skins were analyzed fresh (T0) and at 30 (T1), 60 (T2), 90 (T3), and 120 (T4) days of preservation. Data were analyzed comparatively. There was no bacterial or fungal growth, and the skin structure and collagen arrangement remained intact at all time points in both treatments. In conclusion, both preservation methods are efficient and capable of maintaining the tissue morphological structure and preventing the growth and proliferation of contaminants for up to 120 days.

A utilização de membranas biológicas em curativos tem se tornado cada vez mais uma realidade. Concomitante, busca-se um meio de conservação ideal que possa proporcionar a manutenção do tecido por longos períodos de tempo sem interferir em sua qualidade e aplicabilidade clínica. Desta forma, o objetivo deste trabalho foi o de avaliar e comparar histologicamente e microbiologicamente peles de rã submetidas a dois diferentes métodos de conservação. Foram avaliadas 16 peles de rã-touro as quais foram, em função do método de conservação, subdivididas em dois grupos com 08 peles cada: O Grupo Congelamento (GC) no qual as peles foram submetidas ao congelamento a -4º, em solução de glicerina a 20%; e o grupo Glicerina (GG), no qual as peles foram conservadas em temperatura ambiente (média de 28ºC e umidade média de 78%) em glicerina a 98%. As peles foram analisadas a fresco (T0) e com 30 (T1), 60 (T2), 90 (T3) e 120 (T4) dias de conservação. Os dados foram analisados de forma comparativa. Em todos os tempos analisados e em ambos os tratamentos, não houve crescimento bacteriano ou fúngico e a estrutura da pele e o arranjo de colágeno mantiveram-se íntegros. Conclui-se que, ambos os métodos de conservação são eficientes e capazes de manter a estrutura morfológica tecidual, e impedem o crescimento e a proliferação de contaminantes por até 120 dias.

Glaucoma in dogs and cats - management with cyclocryotherapy

Background: The treatment of glaucoma often requires numerous therapeutic modalities to achieve the desired reduction in intraocular pressure (IOP). Cyclodestructive procedures or ciliary body destruction have been performed using techniques with considerable differences in efficacy and complication rates. Among these methods, cyclocryotherapy is non-invasive and simple for the management of uncontrolled glaucoma in dogs and cats. The objective of this case report is to describe the technique of carbon dioxide cyclocryotherapy to reduce intraocular pressure in dogs and cats with glaucoma. Cases: Nine canine patients and one cat with glaucoma were treated with cyclocryotherapy performed under general anaesthesia. Clinical signs patients included blepharospasm, ocular pain, episcleral congestion and ocular hypertension. The patients showed higher levels of IOP, higher than 30 mmHg. Surgical treatment with general anaesthesia was applied. The pre-anaesthesia protocol included acepromazine 0.05 mg/kg with methadone 0.2 mg/kg, followed by intravenous propofol and maintenance with isoflurane and oxygen. An ophthalmological cryocautery unit was used with carbon dioxide as the cryogenic agent and a retinal cryoprobe of 3.2 mm diameter tip for the procedure. The method used was a double cycle of freezing and thawing for 60 s in each site. The cryoprobe was centred approximately 5 mm posterior to the corneoscleral limbus over the ciliary body. The temperature of each cyclocryotherapy spot was between -60°C and -80°C and each site was maintained in place for 60 s; 4 to 6 spots of the double freeze-thaw cycle were made. This technique did not have any serious complications during or after the application of cryotherapy, but chemosis and hyperaemia of the bulbar conjunctiva developed. Subconjunctival anti-inflammatory steroids were injected to minimise swelling and patient discomfort. Satisfactory results were observed; in all cases, the intraocular pressure decreased, with the usual result being a cosmetic and painless eye. Discussion: Even with recent surgical and medical advances, pain and blindness are still common occurrences in glaucoma in human and veterinary practice. The cyclodestructive procedures included cyclodialysis, cyclodiathermy, cyclocryotherapy, and cyclophotocoagulation. The cryosurgery in veterinary ophthalmology has many indications for the treatment of ocular diseases and is effective at decreasing intraocular pressure in patients with persistent uncontrolled glaucoma. Cyclocryotherapy has been shown to reduce intraocular pressure in dogs, cats, rabbits and humans with normotensive and glaucomatous eyes. The application of a cryoprobe over the ciliary processes results in ablating ciliary tissue so that aqueous humour inflow is reduced to acceptable levels. In the clinical cases evaluated, there was a reduction in intraocular pressure reaching acceptable levels, with the usual result being cosmetic and painless eye. Medical therapy remains the predominant method for treating glaucoma in veterinary patients; therefore, cyclocryotherapy is an effective, simple way to lower IOP and is a reasonable treatment option for glaucoma management. Cyclocryotherapy has shown good results, with a low learning curve and it can also be repeated if necessary.

Effect of sulforaphane on long-term storage of rabbit semen

In this study, it was aimed to determine the effect of sulforaphane (SFN) on rabbit semen cryopreservation. Semen collected from animals was divided into 5 equal volumes as Control, SFN 5 µM, SFN 10 µM, SFN 25 µM and SFN 50 µM groups. Afterwards, semen analyzes were performed. According to our results, there was no statistical difference between the groups at 4°C. However after freezing thawing, the highest total motility, progressive motility and rapid spermatozoa rate was seen in the 10 µM SFN group, while the lowest was observed in the 50 µM SFN group (P<0.05). Static sperm ratio was highest in the 50 µM group, while the lowest was observed in the 10 µM SFN group. When flow cytometry results examined the rate of acrosomal damaged and dead sperm was the lowest in the 10 µM SFN group, a statistical difference was observed between the control group (P<0.05). The highest rate of sperm with high mitochondrial membrane potential was seen in the 5 µM SFN and 10 µM SFN groups. Apoptosis and ROS rates were found to be lower in the experimental groups compared to the control groups (P<0.05). As a result, SFN supplementation at a dose of 10 µM increased the quality of sperm in the freezing and thawing processes of rabbit semen. In conclusion, 10 µM SFN improved the quality of cryopreservation of rabbit semen.(AU)

Addition of orange, pineapple and beet juices as extenders for cryopreservation of ram semen / Adição de sucos de laranja, abacaxi e beterraba em diluidor para criopreservação de sêmen de carneiros

Searching for improvements in semen cryopreservation, natural substances are commonly studied focusing to improve the sperm quality. The aim of this study were evaluated the effect of adding orange, pineapple, and beet juices in different concentrations and combinations to the ram semen cryopreservation extender. Five ejaculates from five adult rams were used. The semen pool was diluted in egg yolk-based extender and mixed with the following 15 treatments (at a final concentration of 400.106 sptz/mL): orange 10% (O10) and 15% (O15); pineapple 10% (P10) and 15% (P15); beet 10% (B10) and 15% (B15); pineapple + orange 10% (PO10) and 15% (PO15); pineapple + beet 10% (PB10) and 15% (PB15); beet + orange 10% (BO10) and 15% (BO15); pineapple + beet + orange 10% (PBO10) and 15% (PBO15); and the control group (CON). Post-thaw in 0.25 mL straws semen quality analysis of cryopreserved semen was performed by CASA and flow cytometry. Analysis of variance (PROC GLM) was carried out and the averages were compared using the SNK test. Pearson's correlation test was also performed. No effect was noted in the addition of juices to the semen extender prior to cryopreservation. Post-thawed, although, statistically similar to the control group, the total motility of the B10 group reached acceptable standards of total motility. In addition, B10 group showed the highest values (p< 0.05) of progressive motility than control group or other treatments. The addition of 10% beet juice to the ram semen extender can improve the cryopreservation of sperm motility.

Em busca de melhorias na criopreservação do sêmen, substâncias naturais são comumente estudadas com o objetivo de melhorar a qualidade do sêmen. O objetivo deste estudo foi avaliar o efeito da adição de sucos de laranja, abacaxi e beterraba em diferentes concentrações e combinações ao diluidor de criopreservação de sêmen ovino. Foram utilizados cinco ejaculados de cinco carneiros adultos. O pool de sêmen foi diluído em diluente à base de gema de ovo e misturado com os seguintes 15 tratamentos (na concentração final de 400x106 sptz/ml): laranja 10% (O10) e 15% (O15); abacaxi 10% (P10) e 15% (P15); beterraba 10% (B10) e 15% (B15); abacaxi + laranja 10% (PO10) e 15% (PO15); abacaxi + beterraba 10% (PB10) e 15% (PB15); beterraba + laranja 10% (BO10) e 15% (BO15); abacaxi + beterraba + laranja 10% (PBO10) e 15% (PBO15); e o grupo controle (CON). Pós-descongelação em palhetas de 0,25 ml a análise da qualidade do sêmen criopreservado foi realizada pelo CASA e citometria de fluxo. A análise de variância foi realizada e as médias comparadas pelo teste SNK. O teste de correlação de Pearson também foi realizado. Nenhum efeito foi observado na adição de sucos ao diluidor de sêmen antes da criopreservação. Após o descongelamento, embora estatisticamente semelhante ao grupo controle, a motilidade total do grupo B10 atingiu padrões aceitáveis de motilidade total. Além disso, o grupo B10 apresentou os maiores valores (p<0,05) de motilidade progressiva que o grupo controle ou os outros tratamentos. A adição de 10% de suco de beterraba ao diluente de sêmen ovino pode melhorar a criopreservação da motilidade espermática.