Utilização de extrato de própolis verde no resfriamento de sêmen equino

Seminal cooling aims to prolong spermatic lifespan and fertilizing capacity due to reduction in energy metabolism of spermatozoa, providing enough period for transportation from the stallion to the mare. Cooling process can result in ultraestructural, biochemical and functional alterations to spermatozoa. The aim of this study was to evaluate the effect of green propolis hydroalcoholic extract (GPHE) added into equine semen extender and stored at 5ºC. For that, semen was collected from five stallions (Crioulo and Quarter horse) in four opportunities (n=20). These ejaculates were submitted to the following treatments: control group (skim milk glucose semen extender) and groups with the addition of green propolis extract in different concentrations (2,5μl/mL; 5μl/mL; 7,5μl/mL; 10μl/mL). Seminal analyses included motility, mitochondrial functionality, integrity of plasma membrane, DNA and acrossome, being used ejaculates with ≥70% of intact cells. Evaluation was performed at 0; 30; 60; 120; 180; 240; 300; 360 and 1440 minutes during cooled storage at 5ºC. Seminal parameters declined according to storage time in all treatments (P<0.05). Sperm motility reached 0% in 360 minutes in the groups with propolis. The addition of green propolis extract into the semen extender maintained structural characteristics of stallion sperm cell, however it was detrimental no functional characteristics.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPESO resfriamento seminal tem como intuito o prolongamento da viabilidade espermática e capacidade fecundante pela redução no metabolismo energético dos espermatozoides, proporcionando período hábil para transporte do sêmen do garanhão até a égua. O processo de resfriamento pode resultar em alterações ultraestruturais, bioquímicas e funcionais na célula espermática. O objetivo deste experimento foi avaliar o efeito do extrato de própolis verde adicionado ao diluente para resfriamento de sêmen equino a 5ºC. Para isso, foram coletados 4 ejaculados de 5 garanhões da raça Crioula e Quarto de Milha (n=20). Estes ejaculados foram submetidos aos seguintes tratamentos: controle (diluente a base de leite desnatado) e grupos com adição de extrato de própolis verde em diferentes concentrações (2,5μl/mL; 5μl/mL; 7,5μl/mL; 10μl/mL). As análises espermáticas incluíram motilidade, funcionalidade mitocondrial, integridade de membrana plasmática, DNA e acrossoma, sendo utilizados ejaculados contendo ≥70% de células íntegras ás 0h. As avaliações foram realizadas ás 0; 30; 60; 120; 180; 240; 300; 360 e 1440 minutos durante o resfriamento a 5ºC. Os parâmetros seminais diminuíram de acordo com o aumento do tempo em resfriamento em todos tratamentos (P<0,05). A motilidade espermática atingiu 0% em 360 minutos nos tratamentos com própolis. A adição de extrato de própolis verde ao diluente de resfriamento seminal manteve as características estruturais da célula espermática equina, no entanto prejudicou as caraterísticas funcionais

Testicular Teratoma in a Unilateral Right-Sided Abdominal Cryptorchid Horse

Background: Cryptorchidism is characterized by the incomplete descent of one or both testicles to the scrotum, being a hereditary alteration and frequently an unilateral condition. Besides the sexual and aggressive behaviour, the retained testicle is commonly located in abdominal cavity, being considered a risk factor for neoplasm development. The most common testicular neoplasm reported in mammalian species are Sertoli cell tumors, Leydig cell tumors, seminomas and teratomas. A presumptive diagnosis of testicular tumor can be achieved by ultrasonography, although the definitive diagnosis is obtained only by histopathology. In this report, we are presenting a of testicular teratoma in an unilateral abdominal cryptorchid horse. Case: A stallion, American Quarter Horse, 3 year-old, was attended and presented right testicle retained and a left testicle in the scrotum. Transrectal palpation was used to identify a round and firm structure, presumably the right testicle, lateral to the urinary bladder and located in the right side of the abdomen.  Further, a transrectal ultrasound examination showed a complex, round mass with irregular edges containing both cystic and solid structures, hypoechoic fluid-filled cavities separated by linear hyperechoic septa. After a clinical examination, the animal was diagnosed with cryptorchidism and was submitted to orchiectomy and cryptorchidectomy by inguinal approach. Surgery was performed under general anesthesia and postoperative care included cold shower, anti-inflammatory and antibiotic therapy. Testicles were surgically removed and further sent for histopathological examination. The visual appearance of the right undescended testicle showed multiple round, cystic, and solid structures on outer surface, while the left descended testicle was apparently normal. The macroscopic evaluation showed that the affected testicle consisted of a firm to soft solid mass with multiple fluid-filled cystic areas. Microscopically, the testicular architecture was replaced by cysts, fibrous tissue, adipose tissue, glandular structures, and foci of calcification. The histology revealed that the retained testicle had a testicular teratoma. Discussion: Reproductive disorders are common in horses and represent a significant part of the equine practitioner routine. Equine cryptorchidism is the most common non-lethal developmental defect of stallions; Surgery is the best treatment, since this alteration is hereditary. Teratomas have been reported more often in cryptorchid testicles, being usually just diagnosed as an incidental finding during surgical procedure. Under field conditions, usually the testicles are not sent for histopathological evaluation and this fact can contribute to underdiagnoses. Ultrasonography allows clinicians to determine testis location as well as morphological changes in the testes, as well as to elaborate a presumptive diagnose of testicular neoplasm. Histopathology is the best exam to achieve definitive diagnoses in the presence of testicular alterations. In our report, diagnosis of testicular neoplasia was not made before surgery and testicular mass was an incidental finding during the pre-surgical examination. Before testicular enlargement or presence of testicular mass, neoplasia should be included in the differential diagnosis. In conclusion, although rare, teratoma should be included in differential diagnoses of retained testicles, especially those with morphological alterations

Epididymal stallion semen submitted to centrifugation with colloid

A coleta de sêmen da cauda do epidídimo é a última oportunidade de obter espermatozoides de garanhões valiosos, sendo que durante a criopreservação, a etapa de centrifugação é considerada um ponto crítico. A principal hipótese é que a centrifugação com coloides pode melhorar a qualidade dos espermatozoides coletados da cauda do epidídimo de garanhões. Para avaliação da hipótese foram realizados dois experimentos. O experimento um teve por objetivo avaliar o efeito da centrifugação com cushion e com coloide em camada única (SLC) na motilidade de sêmen do epidídimo de garanhões após a etapa de centrifugação. O experimento dois teve o objetivo de determinar o efeito da SLC prévio ao congelamento e após o descongelamento. Experimento 1) Oito garanhões foram submetidos à orquiectomia bilateral e o sêmen foi coletado da cauda dos epidídimos (n=16). Após a coleta, as amostras foram submetidas a três protocolos de centrifugação: Convencional (20 minutos a 600xg), cushioned (20 minutos a 900xg) e SLC (20 minutos a 300xg). Os pellets foram ressuspendidos e as amostras foram submetidas à avaliação laboratorial de motilidade e morfologia espermática. Experimento 2) Dez garanhões foram submetidos a orquiectomia bilateral e o sêmen foi coletado da cauda dos epidídimos (n=20). Para criopreservação, as amostras foram submetidas a: centrifugação convencional (20 minutos a 600xg), SLC prévio a criopreservação (SLC-Pre) (20 minutos a 300xg) e SLC após a criopreservação (SLC+) (20 minutos a 600xg seguidos de uma segunda centrifugação descrita após descongelamento). Os pellets foram ressuspendidos em diluente de congelamento, submetidos ao processo de congelamento em nitrogênio líquido e descongelamento. Os grupos de 6 centrifugação convencional e SLC-Pre foram avaliados imediatamente após descongelamento. O grupo SLC+ foi descongelado e submetido à SLC (20 minutos a 300xg) e ressupendido em diluente de congelamento (SLC+F) ou resfriamento (SLC+C). A motilidade total e a motilidade progressiva das amostras foram avaliadas com análise computadorizada do movimento espermático. A morfologia foi avaliada com auxílio de microscópio com contraste de fase. Funcionalidade de mitocôndria, integridade de membrana e DNA foram avaliados com auxílio de microscópio de fluorescência. Os dados foram analisados por estatística descritiva, simple one-way ANOVA e Teste de Tukey. Experimento 1) a motilidade de espermatozoide submetidos à SLC (p0,05) foi superior do que os submetidos a centrifugação convencional. Experimento 2) SLC-Pre e SLC+F apresentaram maior motilidade total, enquanto SLC+F apresentou maior motilidade progressiva. O percentual de espermatozoides com morfologia normal foi maior em SLC-Pre e SLC+F. A funcionalidade de mitocôndria foi maior em todos grupos com SLC, enquanto a integridade de membrana foi maior em SLC-Pre. A centrifugação com coloides melhorou a qualidade de espermatozoides coletados da cauda do epidídimo de garanhões, tanto no momento prévio ao congelamento como após o descongelamento.Epididymis cauda sperm recovery and cryopreservation are last opportunity to obtain spermatozoa from a valuable animal, even though during cryopreservation centrifugation step is considered as a critical point. It is hypothesized that colloidal centrifugation could enhance epididymal stallion sperm parameters. To evaluate this hypothesis two experiments were performed. In experiment one, the objective was to evaluate the effect of cushioned and Single Layer Centrifugation (SLC) on epididymal stallion sperm motility postcentrifugation. In experiment two, the objective was to determine the effect of SLC on epididymal stallion sperm quality pre-freezing and post-thawing. Experiment 1) Eight stallions were submitted to bilateral orchiectomy and the resulting epididymal cauda (n = 16) were flushed with semen extender. After harvesting, samples were submitted to three centrifugation protocols: conventional (20 minutes at 600xg), cushioned (20 minutes at 900xg), and SLC (20 minutes at 300xg). Pellets were resuspended, motility and morphology were evaluated. Experiment 2) Ten stallions were submitted to bilateral orchiectomy and epididymal cauda (n=20) were harvested. For cryopreservation, epididymal sperm were submitted to: conventional centrifugation (20 minutes at 600xg), Single Layer Centrifugation prior cryopreservation (SLC-Pre) (20 minutes at 300xg) and Single Layer Centrifugation after cryopreservation (SLC+) (20 minutes at 600xg followed by a second centrifugation described after thawing). Pellets were resuspended in freezing extender, submitted to cryopreservation process in liquid nitrogen and thawed. Conventional and SLC-Pre were evaluated immediately after thawing. SLC+ samples were thawed, submitted to SLC (20 minutes at 300xg) and the pellets were resuspended with freezing (SLC+F) and cooling extender (SLC+C). Total motility (TM) and progressive 8 motility (PM) were evaluated with computer-assisted semen analyses. Sperm morphology was evaluated under a phase-contrast microscope. Mitochondrial functionality, membrane e DNA integrity were evaluated with an epifluorescence microscope. Data was evaluated by descriptive statistics, simple one-way ANOVA and comparison between means by Tukey test. Significance was assigned to all values p0.05) were higher than those recovered by conventional centrifugation. Experiment 2) SLC-Pre and SLC+F yielded the highest TM, while SLC+F yielded the highest PM. Higher morphological normal sperm was observed in SLC-Pre and SLC+F. Mitochondrial functionality was significantly higher in all treatments with SLC, while membrane integrity was higher in SLC-Pre. Colloidal centrifugation improved epididymal sperm quality before freezing and after thawing

Epididymal stallion semen submitted to centrifugation with colloid

A coleta de sêmen da cauda do epidídimo é a última oportunidade de obter espermatozoides de garanhões valiosos, sendo que durante a criopreservação, a etapa de centrifugação é considerada um ponto crítico. A principal hipótese é que a centrifugação com coloides pode melhorar a qualidade dos espermatozoides coletados da cauda do epidídimo de garanhões. Para avaliação da hipótese foram realizados dois experimentos. O experimento um teve por objetivo avaliar o efeito da centrifugação com cushion e com coloide em camada única (SLC) na motilidade de sêmen do epidídimo de garanhões após a etapa de centrifugação. O experimento dois teve o objetivo de determinar o efeito da SLC prévio ao congelamento e após o descongelamento. Experimento 1) Oito garanhões foram submetidos à orquiectomia bilateral e o sêmen foi coletado da cauda dos epidídimos (n=16). Após a coleta, as amostras foram submetidas a três protocolos de centrifugação: Convencional (20 minutos a 600xg), cushioned (20 minutos a 900xg) e SLC (20 minutos a 300xg). Os pellets foram ressuspendidos e as amostras foram submetidas à avaliação laboratorial de motilidade e morfologia espermática. Experimento 2) Dez garanhões foram submetidos a orquiectomia bilateral e o sêmen foi coletado da cauda dos epidídimos (n=20). Para criopreservação, as amostras foram submetidas a: centrifugação convencional (20 minutos a 600xg), SLC prévio a criopreservação (SLC-Pre) (20 minutos a 300xg) e SLC após a criopreservação (SLC+) (20 minutos a 600xg seguidos de uma segunda centrifugação descrita após descongelamento). Os pellets foram ressuspendidos em diluente de congelamento, submetidos ao processo de congelamento em nitrogênio líquido e descongelamento. Os grupos de 6 centrifugação convencional e SLC-Pre foram avaliados imediatamente após descongelamento. O grupo SLC+ foi descongelado e submetido à SLC (20 minutos a 300xg) e ressupendido em diluente de congelamento (SLC+F) ou resfriamento (SLC+C). A motilidade total e a motilidade progressiva das amostras foram avaliadas com análise computadorizada do movimento espermático. A morfologia foi avaliada com auxílio de microscópio com contraste de fase. Funcionalidade de mitocôndria, integridade de membrana e DNA foram avaliados com auxílio de microscópio de fluorescência. Os dados foram analisados por estatística descritiva, simple one-way ANOVA e Teste de Tukey. Experimento 1) a motilidade de espermatozoide submetidos à SLC (p0,05) foi superior do que os submetidos a centrifugação convencional. Experimento 2) SLC-Pre e SLC+F apresentaram maior motilidade total, enquanto SLC+F apresentou maior motilidade progressiva. O percentual de espermatozoides com morfologia normal foi maior em SLC-Pre e SLC+F. A funcionalidade de mitocôndria foi maior em todos grupos com SLC, enquanto a integridade de membrana foi maior em SLC-Pre. A centrifugação com coloides melhorou a qualidade de espermatozoides coletados da cauda do epidídimo de garanhões, tanto no momento prévio ao congelamento como após o descongelamento.Epididymis cauda sperm recovery and cryopreservation are last opportunity to obtain spermatozoa from a valuable animal, even though during cryopreservation centrifugation step is considered as a critical point. It is hypothesized that colloidal centrifugation could enhance epididymal stallion sperm parameters. To evaluate this hypothesis two experiments were performed. In experiment one, the objective was to evaluate the effect of cushioned and Single Layer Centrifugation (SLC) on epididymal stallion sperm motility postcentrifugation. In experiment two, the objective was to determine the effect of SLC on epididymal stallion sperm quality pre-freezing and post-thawing. Experiment 1) Eight stallions were submitted to bilateral orchiectomy and the resulting epididymal cauda (n = 16) were flushed with semen extender. After harvesting, samples were submitted to three centrifugation protocols: conventional (20 minutes at 600xg), cushioned (20 minutes at 900xg), and SLC (20 minutes at 300xg). Pellets were resuspended, motility and morphology were evaluated. Experiment 2) Ten stallions were submitted to bilateral orchiectomy and epididymal cauda (n=20) were harvested. For cryopreservation, epididymal sperm were submitted to: conventional centrifugation (20 minutes at 600xg), Single Layer Centrifugation prior cryopreservation (SLC-Pre) (20 minutes at 300xg) and Single Layer Centrifugation after cryopreservation (SLC+) (20 minutes at 600xg followed by a second centrifugation described after thawing). Pellets were resuspended in freezing extender, submitted to cryopreservation process in liquid nitrogen and thawed. Conventional and SLC-Pre were evaluated immediately after thawing. SLC+ samples were thawed, submitted to SLC (20 minutes at 300xg) and the pellets were resuspended with freezing (SLC+F) and cooling extender (SLC+C). Total motility (TM) and progressive 8 motility (PM) were evaluated with computer-assisted semen analyses. Sperm morphology was evaluated under a phase-contrast microscope. Mitochondrial functionality, membrane e DNA integrity were evaluated with an epifluorescence microscope. Data was evaluated by descriptive statistics, simple one-way ANOVA and comparison between means by Tukey test. Significance was assigned to all values p0.05) were higher than those recovered by conventional centrifugation. Experiment 2) SLC-Pre and SLC+F yielded the highest TM, while SLC+F yielded the highest PM. Higher morphological normal sperm was observed in SLC-Pre and SLC+F. Mitochondrial functionality was significantly higher in all treatments with SLC, while membrane integrity was higher in SLC-Pre. Colloidal centrifugation improved epididymal sperm quality before freezing and after thawing

Essential basil oil effect (Ocimum basilicum L.) on cattle tick Rhipicephalus (Boophilus) microplus in in vitro experiments<br>Efeito do óleo essencial de manjericão (Ocimum basilicum L.) sobre o carrapato bovino Rhipicephalus (Boophilus) microplus em ensaios in vitro

The excessive use of acaricidal products induces environmental and human contamination, whereas there is also the selection of resistant tick’s strains. Most of the chemical acaricides available in the market have low efficacy due intensive use and inadequate management of treatments. Thus, ecologically viable alternatives for the tick’s control is a promising area of research. Herbal medicine has as fundamental issue the study of medicinal plants and their use in the control and treatment of human and animal diseases. This experiment has evaluated the in vitro efficacy of Ocimum basilicum essential oil on engorded teleogines of Rhipicephalus (Boophilus) microplus. Six concentrations of the oil (1; 5; 10; 25, 50; 100%) were tested, diluted in distilled water, using the engorged female immersion test. The inhibition of oviposition were 10; 26; 26; 33; 100 and 100%, the egg hatch were 75; 60; 80; 45; 0 and 0%, and the effectiveness of treatment 28; 38; 36; 62; 100 and 100%, respectively. The statistic evaluation was done by the linear regression test, and it was observed a significant association between the concentration of the oil and the inhibition of oviposition, egg hatch and effectiveness. The oil at 25% had higher than 50% of effectiveness, and at 50 and 100% had total inhibition of oviposition with maximum efficiency. According to the results, it is possible to conclude that the Ocimum basilicum oil had in vitro acaricidal action on R. (B) microplus teleogines, obtaining partial control of this parasite. O uso excessivo de acaricidas predispõe a contaminação do meio ambiente, do homem e estimula a seleção de cepas de carrapatos resistentes. A maioria dos acaricidas químicos disponíveis no mercado apresenta eficácia reduzida devido ao intenso uso e manejo inadequado dos tratamentos. Sendo assim, alternativas ecologicamente viáveis utilizadas no controle dos carrapatos é uma área de pesquisa promissora. A fitoterapia tem como fundamento o estudo de plantas medicinais, assim como sua utilização no controle e tratamento de enfermidades tanto na área humana quanto na animal. Neste experimento foi avaliado o efeito in vitro do óleo essencial de manjericão (Ocimum basilicum) sobre fêmeas ingurgitadas de Rhipicephalus (Boophilus) microplus. Para isso foram testadas 6 concentrações do óleo de manjericão (1; 5; 10; 25, 50; 100%), diluído em água destilada, seguindo a metodologia do exame de biocarrapaticidograma. Perante os resultados foi observada inibição de postura de 10; 26; 26; 33; 100 e 100%, eclosão larval de 75; 60; 80; 45; 0 e 0% e a eficiência do tratamento de 28; 38; 36; 62; 100 e 100%, respectivamente. A avaliação estatística foi realizada pela análise de regressão linear, onde foi constatada associação significativa entre a concentração do óleo e a inibição de postura, eclosão larval e eficiência do tratamento. O óleo de manjericão á 25% obteve acima de 50% de eficiência, e nas concentrações de 50 e 100% obteve total inibição de postura com consequente eficiência máxima. Conforme os resultados, pode-se concluir que óleo de manjericão apresentou efeito acaricida in vitro sobre as teleóginas de R. (B) microplus, obtendo um controle parcial do parasito

Efeito de diferentes concentrações de óleo e tintura de citronela sobre os parâmetros reprodutivos de teleóginas de Rhipicephalus (Boophilus) microplus

The tick Rhipicephalus (Boophilus) microplus is the most important ectoparasite in cattle rising, and it is responsible for severe economic losses. Parasite control is based on chemicals, which are used indiscriminately and result in effectiveness reduction of these compounds. In order to delay the onset of parasite resistance, some alternative methods are being researched, including herbal medicine. The aim of this experiment was to evaluate the in vitro efficacy of citronella (Cymbopogon nardus) oil and tincture on R. (B) microplus by the engorged female immersion test. Four oil concentrations were tested (1; 25; 50; 100%) and the inhibition of oviposition were 16; 40; 53; 73%, egg hatch were 28; 16; 23; 6% and the effectiveness were 78; 100; 86; 98%, respectively. The gradual rise of the inhibition of oviposition and low egg hatch had a positive influence in the effectiveness of treatment. The solution at 1% of citronella oil had a partial control of the teleogines and the solution at 25% had 100% of effectiveness. Six dilutions of the tincture (1; 5; 10; 25; 50; 100%) were tested and the inhibition of oviposition were 10; 63; 80; 83; 86; 96%, egg hatch were 91; 31; 6; 26; 0; 0% and the effectiveness were 23; 93; 94; 97; 100; 100%, respectively. Most of the treatments with the citronella tincture had inhibition of oviposition higher than 80% and low egg hatch rate. The tincture at 5% had similar efficacy compared to the groups with higher concentrations. A significant association between the oil and tincture of citronella and the treatment effectiveness was observed in the statistical evaluation. It is possible to conclude that both the oil and the tincture of citronella had negative influence in the reproductive parameters of R. (B) microplus.O carrapato Rhipicephalus (Boophilus) microplus é o ectoparasito mais importante em áreas de exploração pecuária, sendo responsável por severas perdas econômicas. O controle deste parasito é feito através de produtos químicos, que são utilizados de forma indiscriminada e resultam na redução da eficiência destes compostos. A fim de retardar o aparecimento da resistência parasitária, alguns métodos de controle alternativo vêm sendo pesquisados, entre eles a fitoterapia. O objetivo deste experimento foi avaliar a eficiência in vitro do óleo e da tintura de citronela (Cymbopogon nardus) sobre fêmeas ingurgitadas de R. (B) microplus seguindo a metodologia do exame de biocarrapaticidograma. Para isto foram testadas 4 diluições do óleo (1; 25; 50 e 100%), sendo que a inibição de postura foi de 16; 40; 53 e 73%, a eclosão larval de 28; 16; 23 e 6% e a eficácia de 78; 100; 86 e 98%, respectivamente. O gradativo aumento da inibição da postura e a baixa eclosão larval influenciaram positivamente na eficiência dos tratamentos. A solução do óleo a 1% apresentou controle parcial das teleóginas e a solução a 25% apresentou 100% de eficiência. Foram testadas 6 diluições da tintura de citronela (1; 5; 10; 25; 50 e 100%), sendo que a inibição da postura foi de 10; 63; 80; 83; 86 e 96%, a eclosão larval de 91; 31; 6; 26; 0 e 0% e a eficácia de 23; 93; 94; 97; 100 e 100%, respectivamente. A maioria dos tratamentos com a tintura de citronela apresentou inibição de postura superior a 80% e baixa taxa de eclosão larval. A tintura a 5% teve eficácia semelhante aos tratamentos com maiores concentrações. Na avaliação estatística foi verificado associação significativa entre a concentração do óleo e da tintura de citronela com a eficiência do tratamento. Conclui-se que tanto o óleo como a tintura de C. nardus demonstraram efeito negativo nos parâmetros reprodutivos do carrapato R. (B) microplus, em experimento in vitro

Posição de cabeça e administração de disofenolato de levamisole em ovinos

A ovinocultura sofre grandes perdas econômicas devido ao parasitismo por nematoides gastrintestinais, sendo que o principal método de controle é através da administração de produtos químicos. Em anti-helmínticos orais, a taxa de absorção sistêmica pode ser alterada por fatores relacionados ao trato gastrintestinal, incluindo a estimulação da goteira esofágica. O objetivo deste experimento foi avaliar a eficiência do disofenolato de levamisole administrado com a cabeça do animal em posição erguida ou alinhada à linha dorsal em ovinos. Para isso foram utilizados 51 ovinos previamente submetidos a um teste de eficácia, onde o disofenolato de levamisole apresentou 99% de eficiência. Os animais foram separados aleatoriamente em três grupos e submetidos a 24h de restrição alimentar prévia a administração do anti-helmíntico. No tratamento 1 (T1), o disofenolato de levamisole foi administrado mantendo a cabeça do animal em posição erguida, acima da linha horizontal do dorso. No tratamento 2 (T2), o mesmo medicamento foi administrado com a cabeça do animal em posição normal, alinhada à linha dorsal. O tratamento 3 (T3) foi o grupo controle, não tratado com anti-helmíntico. Amostras fecais foram coletadas no dia 0, 7 e 14 para realização dos exames de contagem de ovos por grama de fezes (OPG) e coprocultura. Os resultados foram submetidos ao teste estatístico Qui-quadrado. A eficácia no dia 7 e 14 foi de 98% e 100%, respectivamente, tanto para o T1 quanto para o T2. A administração de disofenolato de levamisole com a cabeça do animal em posição erguida ou alinhada à linha horizontal em ovinos submetidos à restrição alimentar não resulta em alteração na eficiência do anti-helmíntico

Water immersion effect on reproductive parameters of Rhipicephalus (Boophilus) microplus ticks

O carrapato Rhipicephalus (Boophilus) microplus é um parasita de destaque na pecuária brasileira devido às perdas econômicas causadas ao rebanho bovino. Este parasito apresenta duas fases de vida, uma fase parasitária que ocorre no corpo do hospedeiro e uma fase de vida livre que ocorre no solo e vegetação. O acúmulo de água no solo influencia na biologia dos ixodídeos, além de outros fatores fundamentais como a temperatura e a umidade relativa do ar. Este experimento teve a intenção de investigar a ação deletéria da imersão em água, em condições controladas, de fêmeas de R. microplus. Para isso, as teleóginas permaneceram imersas em água por 0, 2, 4, 6, 12, 18, 24, 30, 36, 42, 48, 60, 72, 84, 96, 108 e 120 horas conforme tratamento, sendo posteriormente colocadas em uma estufa climatizada para verificação da postura de ovos e eclosão das larvas. Foi observado que em um período de até 30h de imersão, as teleóginas realizaram postura, no entanto mais de 50% ovos não eclodiram. Após 48h, foi observado a postura em mais de 50% das fêmeas e a taxa de eclosão larval foi inferior a 31%. A partir de 84h não houve mais eclosão larval e o tratamento atingiu 100% de eficiência. Com base nos resultados obtidos, conclui-se que a imersão em água por um período superior a 2 horas possui efeito negativo nos parâmetros reprodutivos do carrapato bovino, através de alterações na postura e na eclosão larval.The tick Rhipicephalus (Boophilus) microplus is an important parasite in the cattle raising field due to the economic losses it causes to Brazilian herds. This parasite has two stages of life, a parasitic stage that takes place in the host body and a free-living stage that takes place on soil and vegetation. The water accumulation on the soil has a strong influence in the biology of the Ixodidae, as well as other factors such as temperature and relative humidity. The aim of this experiment was to evaluate the possible deleterious effect of water immersion in controlled conditions of R. microplus females. For that, the teleogines were immersed in water for 0, 2, 4, 6, 12, 18, 24, 30, 36, 42, 48, 60, 72, 84, 96, 108 and 120 hours, according to the treatment. After this part, the ticks were taken to an incubator to finish the egg-laying and hatching. After 30 hours of water immersion, the teleogines had done the egg-laying, however more them half of the eggs did not hatch. After 48h, half of the ticks had done the laying and the hatching rate was lower than 31%. After 84h, there were no more hatching and the treatment had 100% efficacy. After analyzing the results, we can conclude that water immersion for more than 2 hours have a negative effect on cattle ticks reproduction parameters, through alterations in the egg-laying and in the hatching

Cushioned and single layer centrifugation improve epididymal stallion sperm motility postcentrifugation

Traumatic injuries and sudden death can prematurely end the breeding career of a stallion. In such cases, a final spermatozoa collection can be obtained by harvesting the cauda epididymis. Semen samples can then be used for fresh artificial insemination or cryopreserved. Centrifugation is a critical point of sperm cryopreservation processing and can be detrimental to spermatozoa. Colloid centrifugation approaches reduce this physical damage and can be used to select better quality sperm. The aim of this research was to determine the effect of cushioned and single layer centrifugation (SLC) on epididymal stallion sperm motility postcentrifugation. Eight stallions were submitted to bilateral orchiectomy and the resulting epididymal cauda (n = 16) were flushed with semen extender. After harvesting, the samples were submitted to three centrifugation protocols: conventional (20 minutes at 600×g), cushioned (20 minutes at 900×g), and SLC (20 minutes at 300×g). The pellets were resuspended and sperm parameters were evaluated. Sperm morphology was evaluated under a phase-contrast microscope, total motility (TM) and progressive motility (PM) were evaluated with computer-assisted semen analyses. The proportion of morphologically normal spermatozoa was 72.2% for SLC, 72% for cushioned, and 67% for conventional (P > .05). After conventional centrifugation, it was recorded a TM of 7.4% and PM of 2.7%. After cushioned centrifugation, it was recorded a TM of 13.9% and PM of 6.5%. After SLC, it was recorded a TM of 46% and PM of 32.1%. The motility of spermatozoa recovered by SLC (P .05) were higher than those recovered by conventional centrifugation. Colloids, including cushioned and SLC, improved epididymal stallion sperm motility postcentrifugation

Diagnostic Process in a Crioulo Horse with Cushing’s Syndrome

Background: Equine pituitary pars intermedia dysfunction, also known as equine Cushing’s syndrome, is a neurodegenerative disease. An important risk factor for Cushing’s is advanced aging and it is the most common endocrine disorder in older horses. The prevalence in horses aged over 10 and 15 years is reported as 9.3% and 21%, respectively. Due to the slow progressive nature of the disease, seasonal variation in hormone output and overlapping endocrine response to other events, accurate diagnosis is challenging. The diagnosis requires the combination of anamnesis, clinical signs, in addition to laboratory tests results. This study aimed to report Cushing’s syndrome in a Crioulo breed horse focusing on diagnostic methods.Case: A 13-year-old male Crioulo breed, orchiectomized, was attended at the Universidade de Passo Fundo (UPF), in Passo Fundo, RS, Brazil. The owner reported that the animal had progressive weight loss and coat abnormal growth, with curly appearance. From visual inspection, body condition score was 4 (1-9) bulging abdomen was noticed, hirsutism, depression and lethargy. Also, there was a large neoplastic mass on the left side of gluteal region. Later, this mass was classified in histopathological examination as a fibroblastic sarcoid and was treated. The animal presented physical parameters within the physiological limits of the specie. Normochromic normocytic anemia and neutrophilic leukocytosis were reported in the hematologic evaluation. In coproparasitological examination, there were 300 eggs per gram of feaces. Hyperadrenocorticism was suspected in the clinical examination and dexamethasone suppression test was performed to confirm the fact. Basal serum was collected at 17 h (M0) and subsequently 40 µg/kg of dexamethasone was administered intramuscularly. Serum samples were taken after 15 (M15) and 19 (M19) h, resulting in cortisol levels of 1.7 and 1.8 μg/dL, respectively. The M15 and M19 results were above reference values for horses (below 1 μg/dL). Combination of information gathered from anamnesis, clinical examination and dexamethasone suppression test resulted in the definitive diagnosis of hyperadrenocorticism, also known as Cushing’s syndrome. Paliative treatment included shearing all over the body and vitamin supplementation.Discussion: In animals without obvious clinical signs, Cushing's syndrome diagnosis is challenging. The most unique and specific clinical signs are the development of abnormal hair coat, mainly hirsutism, delayed or incomplete shedding, and in aged horse, lightening of coat color. The mechanistic cause of these signs is still barely understood. Cushing's is a collection of syndromes each with a unique set of clinical signs and hormone profiles, which varies according to each individual. Complementary examinations are extremely important and endocrine tests are highly recommended in addition to suggestive findings. However, despite the variety of existing tests, false negatives or false positives can frequently happen. Dexamethasone suppression test is considered the gold standard, well validated, practical and low cost for the diagnosis of this disease. In the present report, the combination of anamnesis (13 years old, weight loss, and abnormal coat), clinical exam (hirsutism) and dexamethasone suppression test (over 1 μg/dL of cortisol 15 h and 19 h after dexamethasone administration) resulted in the definitive diagnosis of Cushing’s syndrome. Measurements of plasma concentrations of cortisol and adrenocorticotropic hormone (ACTH), thyrotropin releasing hormone (TRH) stimulation test, serum insulin concentration and necropsy are other available tests. History, clinical signs and test results are important to achieve the definitive diagnoses, and when possible, it is advisable to perform post-mortem evaluation of the pituitary gland