Influence of thawing temperature on sperm motility, structure, and metabolism of frozen bovine semen / Influência da temperatura de descongelamento na motilidade, estrutura e metabolismo do sêmen bovino congelado

ABSTRACT: The heating rate used during semen thawing plays an important role in reducing structural and functional damage to spermatozoa. In this study, we evaluated the influence of thawing temperature on semen quality, reactive oxygen species (ROS) production, and mitochondrial activity of cryopreserved bovine semen. A total of 195 straws of 0.5 mL from five Holstein Friesian bulls were used (39 straws per bull). Samples underwent 8 to 22 years of storage; they were processed under a standard protocol with tris-egg yolk and stored in liquid nitrogen. Samples were thawed for 30 seconds in a water bath at T1: 36 °C, T2: 38 °C or T3: 40 °C. Sperm motility and kinematics, morphology, structural membrane integrity (SMI), functional membrane integrity (FMI), acrosome integrity (AI), ROS, and mitochondrial membrane potential (ΔΨM) of post-thawing bovine sperm were evaluated. Generalized linear models were fitted to the data. Each model included the effects of bull, storage time, and treatment. The Shapiro-Wilk test was used to assess data normality, and means were compared using the Tukey test. T2 and T3 showed better results for sperm motility and kinematic parameters, SMI (%) (T1 41.9 ± 2.3; T2 45.7 ± 1.9; T3 47.4 ± 2.8), ROS (RFU/min) (T1 0.026 ± 0.007; T2 0.032 ± 0.001; T3 0.031 ± 0.001) and high-ΔΨM (RFU x 103) (67.1± 0,4; 71.3 ± 0.4; 74.2 ± 0.4) (P < 0.05). However, T1 had higher FMI (39.3 ± 2.3) than T2 (34.0 ± 1.9) (P < 0.05), though not significantly (P > 0.05) different from T3 (38.4 ± 2.2). Thawing temperatures of 38 °C and 40 °C increases motility, kinetics, membrane integrity, mitochondrial activity and ROS of cryopreserved bovine semen, compared with more conventional thawing at 36 °C.

RESUMO: A taxa de aquecimento usada durante o descongelamento do sêmen desempenha um papel importante na redução dos danos estruturais e funcionais nos espermatozóides. O objetivo desta pesquisa foi avaliar a influência da temperatura de descongelamento na qualidade do sêmen, produção de espécies reativas de oxigênio (ROS) e atividade mitocondrial do sêmen bovino criopreservado. Foram utilizados 195 palhetas de 0,5 mL de cinco touros Holstein Friesian (39 palhetas por touro). As amostras passaram por oito a 22 anos de armazenamento e foram processadas sob protocolo padrão com Tris-gema de ovo e armazenadas em nitrogênio líquido. As temperaturas de descongelamento foram T1: 36 °C, T2: 38 °C, T3: 40 °C, cada uma por 30 segundos em banho-maria. Pós-descongelamento, a motilidade e cinética dos espermatozoides, morfologia, integridade estrutural da membrana (SMI), integridade funcional da membrana (FMI), integridade acrossomal (AI), ROS e potencial de membrana mitocondrial (ΔΨM) foram avaliados. Modelos lineares generalizados foram ajustados. Cada modelo incluiu os efeitos de touro, tempo de armazenamento e tratamento. A normalidade dos dados foi avaliada pelo teste de Shapiro-Wilk e as médias comparadas pelo teste de Tukey. T2 e T3 apresentaram resultados mais elevados para a maioria dos parâmetros de motilidade e cinemática espermática, SMI (%) (T1 41,9 ± 2,3; T2 45,7 ± 1,9; T3 47,4 ± 2,8), ROS (RFU/min) (T1 0,026 ± 0,007; T2 0,032 ± 0,001; T3 0,031 ± 0,001) e alto ΔΨM (RFU x 103) (67,1 ± 0,4; 71,3 ± 0,4; 74,2 ± 0,4) (P < 0,05). No entanto, T1 apresentou maior FMI (%) (39,3 ± 2,3) em comparação a T2 (34,0 ± 1,9) (P < 0,05), mas não foi diferente do T3 (38,4 ± 2,2) (P > 0,05). Conclui-se que as temperaturas de descongelamento de 38 °C e 40 °C produzem um aumento na motilidade, cinética, integridade de membrana, atividade mitocondrial e ROS do sêmen bovino criopreservado, em comparação com o uso mais convencional de uma temperatura de descongelamento de 36 °C.

Espermiogênese alterada em um touro doador de sêmen: relato de caso / Altered spermiogenesis in a semen donor bull: case report

A espermiogênese é o processo final da espermatogênese no qual a espermátide se transforma em espermatozoide. Durante esse processo podem ocorrer alterações espermáticas, especialmente no acrossoma, na morfologia da cabeça, na condensação da cromatina e na formação dos vacúolos nucleares. A diferenciação entre os quadros clínicos é feita com repetições dos exames andrológicos. A avaliação morfológica indica a situação da espermiogênese nas 4 semanas anteriores a coleta do sêmen. Com os resultados do exame andrológico é possível identificar a qualidade seminal naquele período. O presente artigo é o relato de caso de um touro jovem doador de sêmen com 26 meses de idade, no início do regime de coletas de sêmen em um Centro de Coleta e Processamento de Sêmen no Sul do Brasil. A produção espermática deste animal foi avaliada durante 12 meses de coleta, apresentando sempre morfologia espermática muito alterada e motilidade média de 54% no exame imediato. Em 13 coletas seminais, o ejaculado deste animal apresentou em média 86% de defeitos maiores, 10% de defeitos menores e 96% de defeitos totais. Os defeitos maiores, que são os que possuem maior efeito na fertilidade, estão diretamente ligados a espermiogênese e os defeitos menores, que possuem menor efeito na fertilidade, são ocasionados principalmente durante o trânsito pelo epidídimo. O quadro clínico desse animal demonstrou a importância do exame morfológico para avaliar a espermatogênese e principalmente o processo de diferenciação final da espermátide em espermatozoide. Casos como este devem ser descritos como espermiogênese alterada e o reprodutor deve ser afastado da reprodução, após descartar alterações morfológicas devido a degeneração testicular grave.(AU)

Spermiogenesis is the final process of spermatogenesis in which the spermatid transforms into sperm. During this process, sperm alterations may occur, especially in the acrosome, head morphology, chromatin condensation and formation of nuclear vacuoles. Differentiation between clinical conditions is made with repetitions of andrological exams. The morphological evaluation indicates the status of spermiogenesis in the 4 weeks prior to semen collection. With the results of the breeding soundness examination, it is possible to identify the seminal quality in that period. The present article is a case report of a young 26-month-old semen donor bull, that started semen collection routines at a Semen Collection and Processing Center in southern Brazil. The sperm production of this animal was evaluated during 12 months of collection, always showing very altered sperm morphology and average motility of 54% in the immediate examination. In 13 seminal collections, the ejaculate of this animal presented an average of 86% of major defects, 10% of minor defects and 96% of total defects. Major defects, which have the greatest impact on fertility, are directly linked to spermiogenesis and minor defects, which have less effect on fertility, are mainly caused during transit through the epididymis. The clinical conditions of this animal demonstrated the importance of the morphological examination to evaluate spermatogenesis and especially the process of final differentiation of the spermatid into spermatozoa. Cases like this should be described as altered spermiogenesis and the bull should be withdrawn from breeding, after ruling out morphological changes due to severe testicular degeneration.(AU)

Influence of thawing temperature on sperm motility, structure, and metabolism of frozen bovine semen / Influência da temperatura de descongelamento na motilidade, estrutura e metabolismo do sêmen bovino congelado

The heating rate used during semen thawing plays an important role in reducing structural and functional damage to spermatozoa. In this study, we evaluated the influence of thawing temperature on semen quality, reactive oxygen species (ROS) production, and mitochondrial activity of cryopreserved bovine semen. A total of 195 straws of 0.5 mL from five Holstein Friesian bulls were used (39 straws per bull). Samples underwent 8 to 22 years of storage; they were processed under a standard protocol with tris-egg yolk and stored in liquid nitrogen. Samples were thawed for 30 seconds in a water bath at T1: 36 °C, T2: 38 °C or T3: 40 °C. Sperm motility and kinematics, morphology, structural membrane integrity (SMI), functional membrane integrity (FMI), acrosome integrity (AI), ROS, and mitochondrial membrane potential (ΔΨM) of post-thawing bovine sperm were evaluated. Generalized linear models were fitted to the data. Each model included the effects of bull, storage time, and treatment. The Shapiro-Wilk test was used to assess data normality, and means were compared using the Tukey test. T2 and T3 showed better results for sperm motility and kinematic parameters, SMI (%) (T1 41.9 ± 2.3; T2 45.7 ± 1.9; T3 47.4 ± 2.8), ROS (RFU/min) (T1 0.026 ± 0.007; T2 0.032 ± 0.001; T3 0.031 ± 0.001) and high-ΔΨM (RFU x 103) (67.1± 0,4; 71.3 ± 0.4; 74.2 ± 0.4) (P < 0.05). However, T1 had higher FMI (39.3 ± 2.3) than T2 (34.0 ± 1.9) (P < 0.05), though not significantly (P > 0.05) different from T3 (38.4 ± 2.2). Thawing temperatures of 38 °C and 40 °C increases motility, kinetics, membrane integrity, mitochondrial activity and ROS of cryopreserved bovine semen, compared with more conventional thawing at 36 °C.

A taxa de aquecimento usada durante o descongelamento do sêmen desempenha um papel importante na redução dos danos estruturais e funcionais nos espermatozóides. O objetivo desta pesquisa foi avaliar a influência da temperatura de descongelamento na qualidade do sêmen, produção de espécies reativas de oxigênio (ROS) e atividade mitocondrial do sêmen bovino criopreservado. Foram utilizados 195 palhetas de 0,5 mL de cinco touros Holstein Friesian (39 palhetas por touro). As amostras passaram por oito a 22 anos de armazenamento e foram processadas sob protocolo padrão com Tris-gema de ovo e armazenadas em nitrogênio líquido. As temperaturas de descongelamento foram T1: 36 °C, T2: 38 °C, T3: 40 °C, cada uma por 30 segundos em banho-maria. Pós-descongelamento, a motilidade e cinética dos espermatozoides, morfologia, integridade estrutural da membrana (SMI), integridade funcional da membrana (FMI), integridade acrossomal (AI), ROS e potencial de membrana mitocondrial (ΔΨM) foram avaliados. Modelos lineares generalizados foram ajustados. Cada modelo incluiu os efeitos de touro, tempo de armazenamento e tratamento. A normalidade dos dados foi avaliada pelo teste de Shapiro-Wilk e as médias comparadas pelo teste de Tukey. T2 e T3 apresentaram resultados mais elevados para a maioria dos parâmetros de motilidade e cinemática espermática, SMI (%) (T1 41,9 ± 2,3; T2 45,7 ± 1,9; T3 47,4 ± 2,8), ROS (RFU/min) (T1 0,026 ± 0,007; T2 0,032 ± 0,001; T3 0,031 ± 0,001) e alto ΔΨM (RFU x 103) (67,1 ± 0,4; 71,3 ± 0,4; 74,2 ± 0,4) (P < 0,05). No entanto, T1 apresentou maior FMI (%) (39,3 ± 2,3) em comparação a T2 (34,0 ± 1,9) (P < 0,05), mas não foi diferente do T3 (38,4 ± 2,2) (P > 0,05). Conclui-se que as temperaturas de descongelamento de 38 °C e 40 °C produzem um aumento na motilidade, cinética, integridade de membrana, atividade mitocondrial e ROS do sêmen bovino criopreservado, em comparação com o uso mais convencional de uma temperatura de descongelamento de 36 °C.

Tungíase em cão de uma comunidade rural do estado do Maranhão / Tungíasis in a dog from a rural community in the state of Maranhão

A tungíase é uma infecção parasitária causada pela fêmea da pulga Tunga penetrans, que penetra na pele do hospedeiro para se alimentar e desenvolver seus ovos, gerando lesões que provocam prurido e dor. Assim, o objetivo deste trabalho foi relatar um caso de tungíase em um cão domiciliado em uma comunidade rural de Colinas, Maranhão. Em julho de 2020, um cachorro da raça American Pit Bull Terrier, com um ano e oito meses de idade, foi levado a uma clínica veterinária apresentando irritação e incômodo nas patas, o que dificultava sua locomoção. O cão vivia em uma residência com solo de chão batido, com acesso à rua e o tutor não possuía outros animais. Ao exame físico, constatou-se a presença de ectoparasitos nos coxins palmares e plantares, que foram retirados com auxílio de pinça cirúrgica e encaminhados ao Laboratório de Parasitologia da UFPI para identificação, confirmando tratar-se de pulgas da espécie Tunga penetrans. O cão recebeu tratamento à base de ivermectina 1%, 0,5mL via subcutânea, dose única, apresentando melhora no quadro clínico, e o tutor recebeu orientações sobre a desinfestação do ambiente. Conclui-se que a tungíase pode acometer cães em áreas rurais de um município do Maranhão, sendo importante o diagnóstico precoce e o tratamento adequado.

Tungiasis is a parasitic infection caused by the female flea Tunga penetrans, which penetrates the host's skin to feed and develop its eggs, generating lesions that provoke itching and pain. Thus, this study aimed to report a case of tungiasis in a dog domiciled in a rural community in Colinas, Maranhão. In july 2020, an American Pit Bull Terrier dog, one year and eight months old, was taken to a veterinary clinic with irritation and discomfort in the paws, which made it difficult to move. The dog lived in a house with beaten ground soil, had access to the street, and the tutor had no other animals. The physical examination showed the presence of ectoparasites on the palm and plantar pads, which were removed with surgical tweezers and sent to the UFPI Parasitology Laboratory for identification, confirming that they were Tunga penetrans fleas. The dog received a treatment based on ivermectin 1%, 0.5mL subcutaneously, single dose, showing improvement in the clinical picture, and the tutor received guidance on the disinfection of the environment. It is concluded that tungiasis can affect dogs in rural areas of a city in Maranhão, being important the early diagnosis and adequate treatment.

Seleção de touros para reprodução baseada no fenótipo e DEPs / Selection of bulls for reproduction based on EPD and phenotype

A fertilidade pode ser definida como a capacidade de gerar filhos normais, o que é essencial para o progresso genético, desde que os ascendentes tenham capacidade de transmitir características que venham impactar positivamente os índices zootécnicos e econômicos. Apesar do aumento expressivo do uso de biotécnicas reprodutivas, a maior parte das fêmeas bovinas aptas à reprodução no Brasil ainda são acasaladas por meio da monta natural. Torna-se importante então, não somente, a estimativa da saúde reprodutiva do touro, mas também a avaliação da sua qualidade genética que pode ser feita, por exemplo, pelo acesso os índices dos reprodutores nos diversos programas de melhoramento genético nacional ou pela utilização de escores de avaliação visual.(AU)

Fertility can be defined as the ability to generate normal descendants, which is essential for genetic progress, as long as the ancestors have the ability to transmit characteristics that will positively impact zootechnical and economic indices. Despite the significant increase in the use of reproduction biotechniques, most bovine females capable of reproduction in Brazil are still mated through natural mating. Therefore, it becomes important not only to estimate the reproductive health of the bull, but also to evaluate its genetic quality, which can be done, for example, by accessing the indices of the bulls in the various national genetic improvement programs or by using visual assessment scores.(AU)

Exame clínico andrológico em búfalos: particularidades e mercado / Clinical andrological examination in buffaloes: particularities and market

A bubalinocultura tem expandido mundialmente com grande importância socioeconômica na produção de leite, carne e na tração animal. No entanto, algumas características como: a sazonalidade, poucos reprodutores com genética superior para ser multiplicada e reduzido número de touros com sucesso de coleta constante de sêmen nas Centrais, dificultam a obtenção de maiores índices reprodutivos a nível de Brasil. Neste contexto, a eficiência reprodutiva do rebanho possui papel de destaque, consequentemente, o touro bubalino e a realização do exame andrológico são essenciais para o desenvolvimento da bubalinocultura. Apesar da realização de pesquisas ao longo dos anos relacionadas à andrologia em búfalos, na prática a campo ainda existe uma lacuna referente aos aspectos reprodutivos nessa espécie, fazendo com que haja uma demanda por conhecimento específico. Diante disso, objetivou-se abordar alguns dos principais aspectos da andrologia na espécie bubalina, para auxiliar o Médico Veterinário no atendimento clínico e andrológico visando alcançar uma boa eficiência reprodutiva.(AU)

Buffalo farming has expanded worldwide with great socioeconomic importance in the production of milk, meat and animal traction. However, some characteristics such as: seasonality, few sires with superior genetics to be multiplied and reduced number of bulls with constant semen collection success in the Centrals, make it difficult to obtain higher reproductive rates in Brazil. In this context, the reproductive efficiency of the herd plays a prominent role, consequently, the buffalo bull and the performance of the andrological examination are essential for the development of buffalo breeding. Despite conducting research over the years related to andrology in buffaloes, in field practice there is still a gap regarding reproductive aspects in this species, causing a demand for specific knowledge. In view of this, the objective was to address some of the main aspects of andrology in the buffalo species, to assist the veterinarian in clinical and andrological care in order to achieve good reproductive efficiency.(AU)

Ruptura bilateral do músculo fibular terceiro em bovino / Bilateral rupture of the peroneus tertius muscle in bovine

Background: The peroneus tertius muscle is responsible for tarsal flexion and coordinate extension of the stifle joint. The most common causes of rupture are excessive effort when trying to rise on a slippery surface, mounting, or being mounted. Rupture of the peroneus tertius muscle is characterized by an inability to flex the hock. Most cases are not curable, presenting an unfavorable to poor prognosis, and the animal has to be euthanized. This study aimed to report the clinical picture and treatment of a bilateral rupture of the peroneus tertius muscle in a cow from the municipality of Castanhal, state of Pará (Amazonian biome). Case: We report the clinical picture and treatment of a bilateral rupture of the peroneus tertius muscle in a 4-year-old cow, mixed breed, weighing approximately 400 kg, calved 3 months ago, belonging to a batch of 40 cattle kept on extensive breeding system in a pasture of Urochloa (Brachiaria) brizantha. The clinical signs appeared after the cow was mounted by a 1,100 kg bull and consisted of lameness, falling into sternal decubitus with hind limbs extended backward, hyperextension of the hocks with flexed stifle, with the tibia and metatarsus in a straight line, which lead to a 90-degree position of the femur and knee. Abrasions in the dorsal region of the fetlocks reinforce evidence of dragging hooves. Diagnosis of bilateral rupture of the peroneus tertius muscle was performed by a clinical examination and semiological test. Nonsteroidal anti-inflammatory drugs (flunixin meglumine, 2,2 mg/kg, every 24 h for 4 days) were prescribed for treatment, in addition to resting in a paddock with good availability of pasture, water, and mineral salt. After 3 months, we observed an improvement of the clinical signs, and after 6 months, an almost complete recovery. Only a slight difficulty in flexing the hock when moving remained. Discussion: In the present report, bull mounting was the probable cause of the hyperextension of both hocks and the bilateral rupture of the peroneus tertius muscle. According to the handler, these clinical signs were only evidenced after the animal was mounted. As stated in the literature, accidents such as falls, slips, mounting or being mounted, as well as excessive traction and over-elevation of the limb are considered predisposing factors for the onset of the disease. Rupture of the peroneus tertius muscle is unusual in production animals; therefore, the low occurrence of reports on this kind of rupture in the literature makes this case even more unique. The bilateral injury differs from cases described in the literature, in which the reported injuries were always unilateral. Our findings reinforce the hypothesis that the act of mounting was the triggering factor for the bilateral rupture of the animal peroneus tertius muscle. The breeding system also differs from the cases diagnosed by the same authors, since they describe the disease in bovines kept on intensive or semi-intensive breeding. In the present case, the cattle were kept on an extensive breeding system. The extension and flexion tests used in the clinical examination allowed for the diagnosis of rupture of the peroneus tertius muscle in both limbs, similarly to what has been reported by other authors. Anamnesis, clinical findings, and exclusion of different diagnoses were decisive for establishing the diagnosis. The treatment chosen was efficient, even though the case was severe, as the injury occurred in both hind limbs.

Fertilidade e suscetibilidade à criopreservação espermática: avanços e desafios / Fertility and susceptibility to sperm cryopreservation: advances and challenges

Estudos sobre a fisiologia espermática demonstram que padrões de fertilidade e funcionalidade espermática pós-criopreservação possuem relação importante com a eficiência do metabolismo energético destas células, bem como com a sua capacidade de manter a homeostase oxidativa. Os conhecimentos sobre a relação entre perfil fisiológico de espermatozoides e fertilidade foram e, ainda estão sendo, aprimorados, com o uso de análises de perfis moleculares, com destaque para a metabolômica. As análises moleculares permitiram a identificação de classes de metabólitos importantes na fisiologia espermática, bem como de potenciais biomarcadores de fertilidade, inclusive em bovinos. No entanto, ainda não há disponível uma avaliação isolada capaz de estimar o padrão de fertilidade de amostras seminais. Há vários desafios a serem superados para a validação de biomarcadores de fertilidade, principalmente considerando-se as diferenças entre perfis metabólicos de raças distintas de touros e a heterogeneidade dos ejaculados. A superação destes desafios pode ser iniciada com um maior aproveitamento dos resultados já obtidos e futuros, com a aplicação de análises mais avançadas e com eficácia em elevado número de dados. Para tal, podem ser utilizados modelos estatísticos de inteligência artificial, cuja aplicação pode aumentar a acurácia das observações obtidas, bem como aproximá-las da aplicação pelo setor de produção animal.(AU)

Studies on sperm physiology demonstrate that fertility outcomes and sperm post-cryopreservation have an important relation with sperm energy metabolism efficiency and ability to maintain oxidative homeostasis. Knowledge on sperm physiology has been enhanced, continually, with the application of molecular profiling analysis, focusing on metabolomics. Molecular analysis allowed the identification of important classes of metabolites in sperm physiology, as well as potential fertility biomarkers, including in bovine. Despite all developments, there is still no isolated assessment available capable of estimating fertility on sperm samples. There are several challenges to be overcome for the validation of fertility biomarkers, especially considering the metabolic profiles differences between bull breeds and the ejaculate heterogeneity. Overcoming these challenges could start with the application of more advanced and effective data analysis from research database already obtained, as well as future ones. To this end, statistical models of artificial intelligence can be used, whose application can increase the accuracy of the observations obtained, as well as bringing them closer to the application by the animal production sector.(AU)

Genetic parameters and genetic trends for production traits in dairy Gir cattle / Parâmetros genéticos e tendências genéticas de características de produção de bovinos Gir Leiteiro

The objective of this research was to estimate genetic parameters and genetic trends (GT) for 305-day milk yield (MY305) and 305-day fat yield (FY305) of purebred Dairy Gir animals of the National Dairy Gir Breeding Program. The restricted maximum likelihood method was used in an animal model. GT were obtained via linear regression and divided into two periods (1935-1992 and 1993-2013 for PL305; 1935-1992 and 1993-2010 for MY305). The estimated heritabilities were 0.23 (MY305) and 0.10 (FY305). The GT (kg/year) values for MY305 in the 2nd period for measured females (25.49), females (26.11), and males (35.13) were higher than those found in the 1st period (2.52; 2.06, and 1.00, respectively). The heritability estimated for MY305 confirmed the possibility of genetic improvement by selection and indicated a lower additive genetic effect on FY305 of purebred animals. The genetic progress for MY305 in all purebred population is denoted by the more expressive gains found from 1990's, when the first bull catalogs were published.

Objetivou-se estimar os parâmetros genéticos e tendências genéticas (GT) para produção de leite (MY305) e produção de gordura (FY305), ambas em 305 dias, de animais puros Gir Leiteiro, integrantes do Programa Nacional de Melhoramento do Gir Leiteiro. Foi utilizada a metodologia da máxima verossimilhança restrita em modelo animal. As GT foram obtidas via regressão linear e divididas em dois períodos (1935-1992 e 1993-2013 para PL305; 1935-1992 e 1993-2010 para MY305). As herdabilidades foram de 0,23 (MY305) e 0,10 (FY305). Para PL305, as GT (kg/ano) do 2º período para fêmeas mensuradas (25,49), fêmeas (26,11) e, machos (35,13) foram claramente superiores às do 1º período (2,52; 2,06 e 1,00; respectivamente). A estimativa de herdabilidade para MY305 reafirma ser possível melhoramento genético por meio de seleção, enquanto para FY305 sugere uma menor influência genética aditiva em animais puros. O progresso genético para MY305 em toda a população pura está evidenciado pelos ganhos mais expressivos, observados a partir da década de 90, quando foram divulgados os primeiros sumários de touros.

Gastroesophageal foreign bodies in dogs - endoscopy and surgical removal

Background: Gastroesophageal foreign bodies (GFD) are commonly diagnosed in dogs and are considered an endoscopic emergency that, although not resulting in serious clinical sequelae or mortality, can compromise the health and well-being of the patient. The use of the digestive endoscopy for the diagnosis and treatment of GFD can be a valuable and viable alternative. There are cases of GFD in dogs for which the indicated treatment is surgery, which can be performed using minimally invasive or conventional techniques, associated or not with flexible endoscopy. The objective of this work is to describe 16 cases of GFD removal in dogs demonstrating the efficiency of upper digestive endoscopy. Cases: Of the 16 GFD cases, 63% (10/16) were male and 37% (6/16) female. Most aged under 1 year (63%), puppies (5/16) and juveniles (5/16). The patient with the lowest body weight was a miniature pinscher weighing 0.8 kg (Case 14) and the heaviest was an American Pit Bull Terrier weighing 28 kg (Case 11), the mean body weight of patients diagnosed with GFD was 10.2 ± 6.7 kg. Small and medium breeds were more affected, 44.7% (7/16) and 44.7% (7/16), respectively, and large breeds (Golden Retrievier and Bull Terrier), from cases 1 and 4, the least affected, 12.6% (2/16) of the cases. The 16 patients underwent a 12 h food fast and a 4 h water fast, as gastrointestinal emptying in these cases of GFD can be influenced by these foreign bodies. All underwent general inhalation anesthesia with monitoring of physiological parameters (temperature, heart rate, respiratory rate, oxygen saturation and blood pressure) before, during and after EGD, being positioned in left lateral decubitus. The 16 canine patients with suspected GFD underwent EGD for diagnostic confirmation and removal of foreign bodies. Five esophageal FB were diagnosed, 31% (5/16), and 11 gastric FB, 69% (11/16). The most frequently diagnosed foreign bodies were bone and tissue, 37.5% (6/16) and 31% (5/16). Other foreign bodies were materials such as plastics, metals, rubber, foam and stone. Of the 16 cases of GFD, EGD efficiently treated 88% (14/16) without the need for hospitalization, with only supportive treatment for the remission of complications caused by the presence of foreign bodies in the gastroesophageal tract. The main complications related to the presence of GFD were esophagitis in 25% (4/16) of cases, gastritis in 38% (6/16) and both alterations in 13% (2/16). Discussion: In this work, we can observe that more than a third of the clinical cases of treated dogs were diagnosed with GFD, demonstrating that these cases are common in the veterinary clinic. Most of these animals were males less than 1 year old. The improvement of learning in this category can lead these animals to exacerbated oral exploration of new objects. Most FBs were found in the stomach because they were of adequate size, consistency and shape for their passage through the esophagus, whereas esophageal FBs were all bone fragments of rigid consistency with diameters and sizes larger than the esophageal lumen. The interval between the ingestion of the object and the veterinary care can be decisive for the removal of the FB in the esophagus or stomach. Most gastric FBs removed were fabrics and plastics, flexible objects that can pass through the esophageal lumen more easily. Removal of GFD by endoscopy was performed with a high success rate, with only 2 cases being resolved by esophagostomy and gastrotomy. Flexible endoscopy proved to be an efficient technique for removing treated GFD, which can help remove FB during esophagotomy and be associated with rigid endoscopy. Patients recovered quickly and without complications, but it is important to emphasize that inadequate maneuvers and conducts can determine other outcomes. The use of endoscopy for GFD removal needs to be more popularized, as it can ensure better results for dogs treated with GFD.

Epididymal primary mast cell tumor in a dog

Background: In the literature, there are a few descriptions of epididymis neoplasia in domestic animals, especially considering primary tumors. In the few reports found in literature, the lesions were a consequence of the invasion of testicular or paratesticular neoplasia, as a papillar carcinoma in a dog's and a bull's epididymis, and mesenchymal tumors - fibrome/ fibrosarcoma, leiomyoma/leiosarcome. On the other hand, mast cell tumors are the second most prevalent neoplasia in dogs in Brazil, affecting especially the skin. The aim of this report is to describe for the first time a low malignancy mast cell tumor in a mixed-breed dog's epididymis, without metastasis or recurrence in a 2-year follow-up period. Case: A 10-year-old male mixed-breed dog was presented for pre-surgical evaluation for elective orchiectomy. In the physical examination, an increase in the volume of approximately 2 cm with an irregular appearance was identified on palpation in the cranial pole of the left testis. In the trans surgical period, an increase in testicular volume (4 cm long x 2 cm wide) was observed, with a firm consistency in the region of the vas deferens with macroscopic changes in the region. The testis was sectioned, and the fragments were sent for histopathological evaluation in 10% buffered formaldehyde. There was a fairly cellular circumscribed neoplastic infiltrate, distributed in a sheet and separated by fibrovascular stroma, and rounded neoplastic cells with a moderate amount of basophilic cytoplasmic granulation, and discrete anisocytosis and anisokaryosis. The nuclei were rounded with vesicular chromatin with 1 or 2 distinct nucleoli. No mitosis figures were observed in 10 high power fields (400x). Few eosinophils were distributed throughout the neoplastic cell population. Immunohistochemistry demonstrated immunostaining for KIT protein with perimembranous staining in 95% of neoplastic mast cells, giving a KIT 1 pattern. There was no positive nuclear staining for Ki67 in any cell of the histological sections examined. A grade II mast cell tumor (low grade of malignancy) was diagnosed. After diagnosis, the animal underwent radiographic evaluation of the chest and abdominal ultrasound, and a new physical inspection in search of nodules, plaques, skin lesions, or subcutaneous masses. There were no metastases in the thorax and abdominal cavity, nor physical alterations, and it can be inferred that the epididymis was the primary site of the mast cell tumor. After 2 years of orchiectomy, there were no recurrences, and no chemotherapy treatment was performed. Discussion: Extracutaneous mast cell tumors are uncommon in animals, but have been reported in oral and nasal mucosa, nasopharynx, larynx, trachea, intestine, visceral lymph nodes, spleen, liver, spinal cord, intestine, ureter, conjunctiva, lung and more recently in tear gland of the third eyelid. However, in the authors' assessment, this is the first description of mast cell tumor in the epididymis in dogs. The diagnosis was established by histopathological examination, which revealed a grade II epididymal mast cell tumor and immunohistochemical evaluation (KIT and Ki-67) as being of low aggressiveness. The diagnosis of a primary tumor was confirmed since the staging was established after the histopathological diagnosis, involving chest radiography, abdominal ultrasound, cutaneous evaluation in search of nodules, plaques, cutaneous and subcutaneous lesions, and did not reveal other abnormalities or metastases not identified in the preoperative evaluation. In addition, immunostaining with KIT and Ki-67 reaffirmed the low degree of malignancy and the potential for metastases, which can be observed by the asymptomatic follow-up of the patient 2 years after the surgical excision.

New alternatives for improving and assessing the color of darkcutting beef a review

ABSTRACT Myoglobin (Mb) is a sarcoplasmic heme protein present in muscle cells, which acts as a shortterm oxygen (O2) reserve in the muscle tissue. After slaughtering and exsanguination, Mb is the major pigment that provides the red color in meat. The concentration of Mb together with its redox state are two pivotal factors that determine meat color. The elevated pH of darkcutting beef can affect both physical and biochemical properties resulting in decreased oxygenation. The darkening observed in high ultimate pH (pHu) beef concerns meat processors as color is the initial attribute that impacts on the purchase. Thus, any atypical meat color (i.e., loss of brightness) reduces consumer interest in the product. Several studies have demonstrated that immunological castration is effective in preventing both aggressive behavior and undesirable darkcutting of bull meat. However, little information is available on the effects of processing techniques that limit the oxidation of ferrous iron (Fe2+), Mb or promote metmyoglobin (MMb) reduction in darkcutting beef. Because of the importance of color to fresh beef marketability, this review aimed at overviewing the significance of pHu in beef color and color stability and to discuss new alternatives for improving and assessing the beef color of darkcutting beef, especially in Nellore bulls and their crossbreds, which are widely used in beef cattle production in Brazil.

What is known so far about bull sperm protamination: a review

Sperm routinary fitness evaluation is not sufficient to predict bull reproductive capacity as they present differences in fertility up to 40%. Among the defects which compromise spermatozoa functionality, new approaches consider the study of sperm chromatin, which is the core structure containing paternal genetic information. Sperm chromatin needs to be compacted to maintain the integrity of DNA, which occurs by binding nucleoproteins with high affinity to DNA. In the last stages of sperm maturation, chromatin is hyper-compacted by basic proteins called protamines in a process named protamination. In this review, we summarized intrinsic and extrinsic factors that are suggested to influence protamination in bull spermatozoa, considering old and new evidence from human and murine spermatozoa. Also, the current approaches to evaluate bull protamination and its relationship with fertility were described. Nevertheless, the physiological mechanisms of protamination are still poorly understood.(AU)

Identification of biomarkers for bull fertility using functional genomics

Prediction of bull fertility is critical for the sustainability of both dairy and beef cattle production. Even though bulls produce ample amounts of sperm with normal parameters, some bulls may still suffer from subpar fertility. This causes major economic losses in the cattle industry because using artificial insemination, semen from one single bull can be used to inseminate hundreds of thousands of cows. Although there are several traditional methods to estimate bull fertility, such methods are not sufficient to explain and accurately predict the subfertility of individual bulls. Since fertility is a complex trait influenced by a number of factors including genetics, epigenetics, and environment, there is an urgent need for a comprehensive methodological approach to clarify uncertainty in male subfertility. The present review focuses on molecular and functional signatures of bull sperm associated with fertility. Potential roles of functional genomics (proteome, small noncoding RNAs, lipidome, metabolome) on determining male fertility and its potential as a fertility biomarker are discussed. This review provides a better understanding of the molecular signatures of viable and fertile sperm cells and their potential to be used as fertility biomarkers. This information will help uncover the underlying reasons for idiopathic subfertility.(AU)

Evaluation of novel SexedULTRA-4M technology for in vitro bovine embryo production

SexedULTRA-4M™ is made using an improved method of sex-sorting sperm in a less damaging environment for better retaining sperm integrity throughout the sorting process. The objective of this research was to compare conventional (CONV) and SexedULTRA-4M™ (ULTRA-4M) semen for bovine IVP using four Angus bulls. Matured slaughterhouse oocytes (n = 4000) were divided into the CONV group and the ULTRA-4M group (2000 COCs for each semen type). The IVF process was implemented with CONV and ULTRA-4M semen from the same bull. The cleavage rates, eight cell embryos and blastocysts on day 7 of culture were evaluated for each semen type and each bull. The statistical analysis was carried out with the ANOVA procedure SAS software. The results were 54.45% ± 1.03 and 58.10% ± 1.07; 35% ± 1.57 and 39.15% ± 1.62; 22.8% ± 1.09 and 27.15% ± 1.12 for CONV and ULTRA-4M, respectively, for cleavage rate, eight cell embryos and blastocysts on day 7 for the average of all bulls, comparing only the semen type. Concerning only the semen type, ULTRA-4M was significantly superior to CONV for cleavage rates (P = 0.01) and blastocysts on day 7 (P = 0.009). There were no significant differences between the CONV and ULTRA-4M groups (P>0.05) for all variables analyzed for Bull 1 and Bull 4, however, for Bull 2 ULTRA-4M was significantly superior to CONV for cleavage rates and blastocysts on day 7 (P< 0.05). In Bull 3, ULTRA-4M was significantly higher (P< 0.05) for blastocysts on day 7 compared to CONV. In conclusion, under the conditions of this research the ULTRA-4M and CONV semen produced similar bovine IVP results overall.(AU)

Physiological effects of land and water treadmill exercise in dogs / Efeitos do exercício em esteira terrestre ou aquática sobre os parâmetros fisiológicos de cães

The application of hydrotherapy is growing in the field of animal rehabilitation to improve strength, endurance, fitness, and range of motion, in addition to reducing pain. In the same way, land-based activity is recommended for various conditions, including hypertension, muscle atrophy, and obesity. There is a lack of studies that evaluate the safety of incremental exercise in American Pit Bull Terriers. This study assessed the influence of activity on underwater and dry land treadmills on the physiological parameters of dogs. In this regard, 12 adult, male, healthy, and untrained American Pit Bull Terriers were subjected to an incremental exercise test (IET) on the underwater treadmill (Group I) and the dry land treadmill (Group II). Heart rate (HR), respiratory rate (RR), rectal temperature (RT), systolic blood pressure (SBP), and electrocardiographic evaluation, were evaluated before activity (T0), immediately after (T1), 30 min after (T2), and 90 min after the end of exercise (T3), and plasmatic lactate concentrations were measured at T0, T1, and T2 time points. Results obtained were submitted to the Shapiro-Wilk test, ANOVA and Tukey's test, considering P<0.05. Water activity induced a reduction in RR (P=0.00674) in all examinations after the test on the underwater treadmill and proved to be more demanding than activity on the land treadmill, considering the presence of a deeper and slower RR. Exercise sessions in both groups did not elevate the lactate concentration. The IET can be performed safely on an underwater or dry land treadmill in healthy American Pit Bulls.

A hidroterapia apresenta crescente aplicação no ramo da reabilitação animal, com o intuito de aumentar a força, resistência muscular e amplitude de movimento articular, além de reduzir a dor e melhorar o condicionamento físico do paciente. Assim como a hidroterapia, a atividade desenvolvida em solo é indicada para diversas afecções, inclusive para pacientes hipertensos, com atrofia muscular ou com sobrepeso. São escassos os estudos que avaliam a segurança de testes incrementais em esteira aquática e em esteira seca em American Pit Bull Terriers. Este estudo teve como objetivo avaliar os efeitos da atividade em esteira aquática e em esteira seca sobre os parâmetros fisiológicos de cães. Para isto, cães da raça American Pit Bull Terrier (n=12) machos, adultos, saudáveis e não treinados foram submetidos a teste de esforço incremental (TEI) em hidroesteira com água (Grupo I) e sem água (Grupo II). A frequência cardíaca (FR), frequência respiratória (FR), temperatura retal (TR), pressão arterial sistólica (PAS) e avaliação eletrocardiográfica, foram aferidas antes da atividade (T0), imediatamente após (T1), 30 minutos após (T2) e 90 minutos após o exercício (T3), e a concentração plasmática de lactato foi dosada nos momentos T0, T1 e T2. Os resultados foram submetidos ao teste de Shapiro-Wilk, análise de variância (ANOVA) e pelo Teste de Tukey, considerando significativo o valor de P < 0.05. Houve diminuição da frequência respiratória (P = 0,00674) após a atividade em esteira aquática, que se mostrou mais exigente que a atividade em esteira terrestre, considerando a apresentação de padrão respiratório mais lento e profundo. As sessões de exercício em ambos os grupos não elevaram a concentração de lactato. O teste de esforço incremental em esteira aquática, ou em esteira seca, pode ser desenvolvido com segurança por cães American Pit Bull saudáveis.

Primary adipsia and severe hypernatremia in a Pit Bull dog / Adipsia primária e hipernatremia severa em um cão Pit Bull

Primary adipsia is a rare condition in which there is failure in the activation threshold of the hypothalamic osmoreceptors, leading to osmolality imbalance. Here, we reported the case of a Pit Bull dog with an altered level of consciousness (started after weaning) and adipsia. There was an increase in plasma osmolality (444 mOsm/kg), sodium (223.7 mg/dL), and chlorine (173 mg/dL) levels. Based on the suspicion of primary adipsia, water was administered via a nasogastric tube, with clinical improvement. The owner was instructed to supply water with food. Eight months after discharge, the dog returned with parvovirus and died. In the anatomopathological examination, no structural changes were observed in the central nervous system. To our knowledge, this is the first report of hypernatremia due to adipsia in a Pit Bull dog, showing that this is a differential diagnosis that should be considered in this breed.

Adipsia primária é uma rara condição em que há falha no limiar de ativação dos osmoreceptores hipotalâmicos, levando ao desequilíbrio da osmolalidade. Este artigo tem como objetivo relatar o caso de um cão da raça Pit Bull apresentando alteração no nível de consciência (iniciado após o desmame) e adipsia. Foi verificado aumento da osmolalidade plasmática (444 mOsm/kg), sódio (223,7 mg/dL) e cloro (173 mg/dL). Baseado na suspeita de adipsia primária, iniciou-se administração de água via sonda nasogástrica, com melhora clínica. O tutor foi orientado a fornecer água junto a alimentação. Oito meses após alta, o paciente retornou com parvovirose e veio a óbito. No exame anatomopatológico, não foram observadas alterações estruturais no sistema nervoso central. Este é o primeiro relato de hipernatremia por adipsia em um cão Pit Bull, mostrando que este é um diagnóstico diferencial que deve ser considerado nesta raça.

New alternatives for improving and assessing the color of dark–cutting beef – a review

Myoglobin (Mb) is a sarcoplasmic heme protein present in muscle cells, which acts as a short–term oxygen (O2) reserve in the muscle tissue. After slaughtering and exsanguination, Mb is the major pigment that provides the red color in meat. The concentration of Mb together with its redox state are two pivotal factors that determine meat color. The elevated pH of dark–cutting beef can affect both physical and biochemical properties resulting in decreased oxygenation. The darkening observed in high ultimate pH (pHu) beef concerns meat processors as color is the initial attribute that impacts on the purchase. Thus, any atypical meat color (i.e., loss of brightness) reduces consumer interest in the product. Several studies have demonstrated that immunological castration is effective in preventing both aggressive behavior and undesirable dark–cutting of bull meat. However, little information is available on the effects of processing techniques that limit the oxidation of ferrous iron (Fe2+), Mb or promote metmyoglobin (MMb) reduction in dark–cutting beef. Because of the importance of color to fresh beef marketability, this review aimed at overviewing the significance of pHu in beef color and color stability and to discuss new alternatives for improving and assessing the beef color of dark–cutting beef, especially in Nellore bulls and their crossbreds, which are widely used in beef cattle production in Brazil.

New alternatives for improving and assessing the color of dark­cutting beef ­ a review

Myoglobin (Mb) is a sarcoplasmic heme protein present in muscle cells, which acts as a short­term oxygen (O2) reserve in the muscle tissue. After slaughtering and exsanguination, Mb is the major pigment that provides the red color in meat. The concentration of Mb together with its redox state are two pivotal factors that determine meat color. The elevated pH of dark­cutting beef can affect both physical and biochemical properties resulting in decreased oxygenation. The darkening observed in high ultimate pH (pHu) beef concerns meat processors as color is the initial attribute that impacts on the purchase. Thus, any atypical meat color (i.e., loss of brightness) reduces consumer interest in the product. Several studies have demonstrated that immunological castration is effective in preventing both aggressive behavior and undesirable dark­cutting of bull meat. However, little information is available on the effects of processing techniques that limit the oxidation of ferrous iron (Fe2+), Mb or promote metmyoglobin (MMb) reduction in dark­cutting beef. Because of the importance of color to fresh beef marketability, this review aimed at overviewing the significance of pHu in beef color and color stability and to discuss new alternatives for improving and assessing the beef color of dark­cutting beef, especially in Nellore bulls and their crossbreds, which are widely used in beef cattle production in Brazil.(AU)

Penile fractures in young bulls raised in feedlots in Southern Brazil

Background: Penile fracture is a pathology of young cattle that perform precocious and disordered breeding. The incompatibility of height between males and females and sodomy between males cause a great pressure on the sigmoid flexure and retractor muscle of the penis, which are the main causes and sites of organ injury. This study aimed to describe the epidemiological and pathological aspects of penile fractures observed in young bulls raised in pre-export feedlots (PEFs) in southern Brazil. Cases: In 2 PEFs located in the municipalities of Pelotas (property 1) and Capão do Leão (property 2), 3 male cattle [1 from property 1 and 2 from property 2] presented subcutaneous edema in the foreskin and perineum, associated with dysuria. The evolution of the clinical picture was approximately 20 days in all cases, with evolution to death. The bovine necropsied on property 1 had an increased volume and inguinal edema, involving the penis and scrotal sac. Necrosis of the subcutaneous tissue and local musculature was also observed. The testicles were surrounded by the necrotic tissue, and the right testis was swollen, with flaccid parenchyma adhering to the tunica albuginea. In the necropsy of 1 bull from property 2, an increase in the inguinal volume was observed, with an extensive area of necrosis and edema extending from the prepuce to the caudal musculature of the scrotal sac. There were also marked varicosis in the sigmoid flexure and necrosis of the adjacent region, without the involvement of the corpus cavernosum. During the necropsy of the 2 young bulls, fragments of organs from the abdominal, thoracic, and brain cavities were collected and fixed in 10% buffered formalin. From the bull of the property 2, an anatomical piece consisting of the penis, prepuce, and testicles was also collected and fixed in 10% buffered formalin. After 48 h, the tissue samples were cleaved, embedded in paraffin, cut into 3-µm-thick sections, and stained using hematoxylin and eosin (HE). A histological evaluation of the penile lesions in both cattle revealed intense hemorrhage, congestion, and necrosis of the muscles and tissues adjacent to the corpus cavernosum. In addition to areas of dystrophic calcification, neutrophil and macrophage infiltration was also observed. In the bull from the property 1, an intense edema and proliferation of fibrous tissue surrounding the urethra were noted. There were also marked tubular degeneration and intense infiltration of neutrophils, lymphocytes, and macrophages in the inner portion of the tunica albuginea. Discussion: In the present cases, the diagnosis was based on epidemiological data associated with clinical signs and pathology. The macroscopic lesions observed were probably due to the involvement of blood vessels adjacent to the penis, which suffered trauma during sodomy mating among cattle. These lesions have been described in other reports of this pathology and in diseases, such as acropostitis-phimosis, fibropapilloma of the glans, preputial abscess, and urolithiasis, and the differential diagnosis of these diseases must be carried out, as they have different etiologies. In the bulls of the present study, no lesions were observed in the corpus cavernosum, and this condition was attributed to the presence of varicosis and accumulation of urine in the prepuce, due to the difficulty in exposing the penis. Histologically, there were intense hemorrhage, congestion, and necrosis of the muscles and tissues adjacent to the corpus cavernosum, with the infiltration of neutrophils and macrophages, and areas of dystrophic calcification. The presence of necrotic lesions in tissues adjacent to the penis may be related to hypoxia, vascular lesions, or the action of chemical elements present in the urine. In both cases, vascular lesions were present, which were attributed to the main triggering factor for the disease.