Evaluation of pharmacological alternatives to reduce the pain and discomfort produced by electroejaculation in rams.

Electroejaculation (EE) represents the main technique for semen collection from domestic and wild animals independently of libido. However, the technique is associated with intense involuntary muscle contractions, vocalization, ataxia and lying down, caused by the electric stimulation of the nerves in the caudal epigastric region. These clinical manifestations represent important indicators of discomfort. In this context, the objective of this study was to evaluate two protocols of local anaesthetic blockade and two anatomical access for pharmacological desensitization of the caudal epigastric innervation as alternatives to promote comfort and reduce stress associated with EE in rams. For the study, four clinically healthy Dorper rams were selected. All animals were subjected to a design consisting of five semen collection treatments (n = 3 collections per treatment): T1-control, conventional EE without local anaesthetic blockade; T2, EE with ventral blockade (VB) of epigastric innervation using lidocaine hydrochloride 2%; T3, EE with VB of epigastric innervation using a combination of lidocaine hydrochloride 2% and fentanyl citrate; T4, EE with blockade of epigastric innervation through the perineal access using lidocaine hydrochloride 2%; T5, EE with blockade of epigastric innervation through the perineal access using a combination of lidocaine hydrochloride and fentanyl citrate. Seminal samples resulting from EE were subjectively evaluated for sperm motility and concentration, vigour and volume. Additionally, blood serum samples were collected for quantification of cortisol and creatine kinase (CK) enzyme. Assessments of stress and discomfort were conducted by measuring blood pressure, heart rate (HR) and respiratory rate (RR), as well as observing involuntary muscle contractions, ataxia and animal vocalization. No variations in blood pressure, sperm motility, vigour, CK, and cortisol were observed among the treatments. Individual variations were observed for the occurrence of vocalization (p = .0066), but there were no differences between the groups. Anaesthetic blockades conducted using the combination of lidocaine and fentanyl resulted in a lower incidence of ataxia during EE (p < .0001). It is concluded that the combination of fentanyl citrate and lidocaine hydrochloride results in less discomfort for animals undergoing EE, regardless of the anatomical access used for local anaesthetic blockades.

Collection, Handling, and Preservation of Wild Bird Semen: Current Status, Challenges, and Perspectives.

Semen preservation is a significant biotechnology used to safeguard the genetic material of birds, especially those with declining populations, through biobanking. However, there are limited reports on the successful chilling or cryopreservation of wild bird semen. In general, these techniques are not yet well-established for several species of wild birds and pose several challenges such as the need for bird handling and training, contamination of semen samples, low volume of semen collected, and inefficient preservation protocols. To address these challenges and improve post-thawing outcomes, new possibilities are being investigated, including alternative collection methods to traditional digital massage, the use of antioxidants and enzymes in the medium for chilling or freezing, storage methods using different straws from the usual pellet, and slower freezing rates. This review aims to discuss the various aspects of applying semen preservation in wild birds to create germplasm banks, highlighting the primary results obtained and the challenges that need to be addressed.

Particularidades da reprodução de aves silvestres / Particularities of wild birds reproduction

O conhecimento sobre a reprodução de aves silvestres está em constante evolução com o avanço de novas pesquisas. O número gigantesco de espécies de aves e a dificuldade de acesso aos seus gametas são aspectos complicadores para o estabelecimento de técnicas de reprodução assistida. Aspectos muitos básicos como métodos de coleta de sêmen e as características seminais da grande maioria das aves silvestres são desconhecidos. Essas informações são fundamentais para entender a biologia reprodutiva e planejar programas de multiplicação com o uso de inseminação artificial. Por isso o levantamento das principais particularidades da reprodução de aves silvestres norteará ações para o aprofundamento de pesquisas já realizadas e estímulo ao desenvolvimento de novas metodologias de coleta de sêmen e avaliação seminal em aves silvestres até o momento não estudadas.(AU)

The knowledge about the reproduction of wild birds is constantly evolving with the advancement of new research. The large number of bird species and the difficult access to their gametes are complicating aspects for the establishment of assisted reproduction techniques. Very basic aspects such as semen collection methods and the seminal characteristics of the vast majority of wild birds are unknown. This information is fundamental for understanding reproductive biology and planning multiplication programs using artificial insemination. Therefore, the survey of the main particularities of the reproduction of wild birds will guide actions for the deepening of research already carried out and encourage the development of new methodologies for semen collection and seminal evaluation in wild birds that have not been studied so far.(AU)

Using transrectal ultrasound-guided massage of the accessory sex glands for buck semen collection yields semen with greater cryoresistance than electroejaculation alone during the breeding season.

The cryoresistance of bucks' semen collected by transrectal ultrasound-guided massage of the accessory sex glands (TUMASG) and electroejaculation (EE) was compared during the breeding season (Study 1) or the late non-breeding season (Study 2). Semen was collected from 10 Gabon bucks with both methods in each season and then frozen according to a standardized protocol. The time required for ejaculation, the number of electrical pulses applied, the sum of the electrical pulses applied∗voltage and the total number of vocalizations emitted during the collection by each animal were recorded. In Study 1, TUMASG required a longer time (P = 0.0006) but fewer electrical pulses and a lower sum of pulses∗voltage than EE (P < 0.0001 for both comparisons). Bucks vocalized fewer times during TUMASG than during EE (P < 0001). Semen collected with TUMASG had greater sperm concentration, sperm mass motility, total number of motile sperm and of sperm with progressive motility, and tended to have greater total number of sperm with functional membrane than semen collected with EE (P = 0.02; P = 0.003; P = 0.02; P = 0.02; P = 0.06, respectively). After the freezing-thawing process, sperm collected with TUMASG had a greater quality of the motility, a total number of motile sperm and sperm with progressive motility, a greater percentage of motile sperm and sperm with functional membrane, and tended to have a greater total number of sperm with normal morphology than semen collected with EE (P = 0.04; P = 0.04; P = 0.03; P = 0.02; P = 0.04; P = 0.06, respectively). In Study 1 sperm collected with TUMASG had greater cryoresistance for almost all the variables considered than when it was collected with EE. In Study 2, the number of electrical pulses and the sum of pulses∗voltage were greater with EE than with TUMASG (P < 0001; P = 0.0002, respectively). There were no differences in any sperm variable in fresh or thawed samples collected with TUMASG or EE. However, the other seminal characteristics evaluated did not differ according to the method, and there were no differences in the cryoresistance of any variable. Overall, TUMASG affected animal welfare less than EE and was suitable for collecting semen of good quality with high cryoresistance during the breeding season in buck.

Técnicas alternativas para colheita de sêmen de cão / Alternative dog semen collection techniques

Métodos eficazes para a colheita de sêmen permitem avanços na inseminação artificial em cães domésticos e canídeos selvagens geneticamente valiosos, permitindo a manutenção de linhagens genéticas e possibilitando a propagação de material genético post-mortem, bem como, diminuindo os custos e os riscos de transporte de animais para reprodução. Dentre os métodos de colheita de sêmen utilizados em cães, podemos destacar os métodos ejaculatórios como a manipulação digital e a eletroejaculação; e os métodos não ejaculatórios, que compreendem aqueles que envolvem os espermatozóides coletados do epidídimo, em casos de óbito ou orquiectomia ou em animais submetidos a vasectomia. Mais recentemente a colheita farmacológica de sêmen foi descrita em cães como um método não ejaculatório eficiente. Essa nova metodologia de colheita de sêmen surge como mais uma alternativa para facilitar a colheita de sêmen em cães domésticos que não aceitam a manipulação digital e eliminar a necessidade da eletroejaculação em animais selvagens. No entanto, os protocolos anestésicos utilizados ainda apresentam alguns efeitos colaterais indesejáveis, indicando que novos protocolos, mais seguros para a colheita de sêmen via cateterismo uretral necessitam ser testados.

Effective methods for semen collection are determinant for artificial insemination advances in genetically valuable domestic dogs and wild canids, allowing the genetic maintenance and enabling the post-mortem genetic material use, as well as, reducing the animals transporting costs and risks for breeding. The semen collection methods used in canids are ejaculatory methods, such as digital manipulation in dogs and electroejaculation in wild canids; and non-ejaculatory methods, which include sperm collected from epididymis after death or vasectomy. More recently, pharmacological semen collection was described in dogs as an efficient non-ejaculatory method. This new semen collection methodology emerges as another alternative to facilitate semen collection in domestic dogs that do not accept digital manipulation and eliminate the electroejaculation in wild animals. However, side effects undesirable was observed indicating that new anesthetic protocols, safer for semen collection via urethral catheterization, need to be tested.

Collection of semen in a dog with partial penectomy followed by successful artificial insemination: case report / [Coleta de sêmen em um cão parcialmente penectomizado seguida de inseminação artificial bem sucedida: relato de caso]

This study describes a case of a dog with a lacerated penile tear treated with partial penectomy followed by successful semen collections for artificial insemination. A 1.5-year-old Jack Russel Terrier with normal libido, genital organs and semen, had a penile laceration after copulation. The dog underwent a partial penectomy without orchiectomy, thus preserving the possibility of semen collection. Semen was successfully collected at 45 and 53 days after surgery, and it was used for artificial insemination of two bitches, one of which became pregnant. Therefore, this report demonstrated that semen may be collected from dogs with partial penectomy for artificial insemination, this technique has the potential to preserve fertility of dogs with penile lesions that require penectomy.(AU)

Este estudo descreve o caso de um cão que teve laceração peniana tratada por penectomia parcial seguida de coleta de sêmen para inseminação artificial bem sucedida. Um cão Jack Russel Terrier de 1,5 anos, com libido, órgãos genitais e sêmen normais, teve laceração peniana após cópula. O cão foi parcialmente penectomizado sem orquiectomia, preservando a possibilidade de coleta de sêmen. A coleta de sêmen foi bem sucedida 45 e 53 dias após a cirurgia, sendo utilizado para inseminação artificial de duas cadelas, uma delas se tornando gestante. Portanto, sêmen para inseminação artificial pode ser coletado em cães parcialmente penectomizados, esta técnica revelando ser uma potencial forma de preservação da fertilidade de cães com lesões penianas que necessitem penectomia.(AU)

Semen collection, sperm characteristics and ultrasonographic features of reproductive tissues in crab-eating fox (Cerdocyon thous).

Our objective was to condition adult male crab-eating fox (Cerdocyon thous) to allow semen collection by digital manipulation of the penis, assess sperm characteristics, and use ultrasonography to assess and measure testes and prostate and determine echotexture, as well as hemodynamics of testicular and spermatic cord arteries. Five captive C. thous (2-5 y, ∼7 kg) were used. After conditioning for digital sperm collection, 13 ejaculates were obtained from two males. Ejaculate characteristics (mean ± SD) were volume (393.2 ± 178.0 µL), concentration (463.7 ± 594.4 × 106 sperm/mL), total sperm number (217.4 ± 304.0 × 106 sperm), total motility (68.0 ± 6.1%), progressive motility (48.8 ± 5.1%) and rapid sperm (57.2 ± 6.8%). The most prevalent sperm morphological defects were abnormal acrosome, small abnormal head, and diadem defect. Under chemical restraint, testes and prostate were evaluated with B-mode and spectral Doppler ultrasonography. Based on ultrasonography, testicular volumes were (2.2 ± 0.7 and 2.1 ± 0.7 cm3 for left and right testes, respectively), whereas prostate dimensions were 2.3 ± 0.4 cm long and 1.5 ± 0.3 cm thick. Lastly, using triplex Doppler, peak systolic velocity (PSV) was higher in testicular versus intratesticular arteries. In conclusion, adult male C. thous were conditioned to allow sperm collection by digital manipulation without urine contamination, enabling evaluation of sperm characteristics. Furthermore, B-mode ultrasound facilitated evaluation of testes and prostate morphometry and echotexture, whereas Doppler was used to characterize testicular blood flow. This was proof of concept for semen collection and baseline data for future male reproductive soundness examinations and development of assisted reproductive techniques in this species.

Artificial insemination in rabbits: factors that interfere in assessing itsresults

Understanding the rabbit's sexual behavior andreproduction process is very important when using artificial insemination (AI) instead of natural mating in intensive rearing systems. Ovulation in rabbit females not occur spontaneously/ induced ovulatory animals, but it has to be induced through a neuro-hormonal reflex, which is stimulated during mating. The mating effect of satiety on specific measures of rabbit female sexual behavior has not beeninvestigated. When applying AI, in the absence of a male, ovulation has to be induced by exogenous hormonal analogs. Bio-stimulation techniques are suggested as a substitute for hormones in that respect. One of the most effective approachesto distribute ejaculates from bucks of superior quality or genetic value is the systematic use of AI in intensive rabbit production. Rabbit's sexual receptivity has a significant influence on fertility at insemination. In addition, a limitationfactor for rabbit AI spread is related to their semen preservation. Fresh diluted semen has been used but its quality can be maintained only for a short period. However, there are few articles available have indicated the negativity of AI usage on rabbit, and the factors that can influence the AIprocess including the sexual desire behavior. Therefore, the objective of this review is to identify the important factors that directly or indirectly affect AI process success, meanwhileinterfering with and/or relying on AI assessment.(AU)

Artificial insemination in rabbits: factors that interfere in assessing itsresults

Understanding the rabbit's sexual behavior andreproduction process is very important when using artificial insemination (AI) instead of natural mating in intensive rearing systems. Ovulation in rabbit females not occur spontaneously/ induced ovulatory animals, but it has to be induced through a neuro-hormonal reflex, which is stimulated during mating. The mating effect of satiety on specific measures of rabbit female sexual behavior has not beeninvestigated. When applying AI, in the absence of a male, ovulation has to be induced by exogenous hormonal analogs. Bio-stimulation techniques are suggested as a substitute for hormones in that respect. One of the most effective approachesto distribute ejaculates from bucks of superior quality or genetic value is the systematic use of AI in intensive rabbit production. Rabbit's sexual receptivity has a significant influence on fertility at insemination. In addition, a limitationfactor for rabbit AI spread is related to their semen preservation. Fresh diluted semen has been used but its quality can be maintained only for a short period. However, there are few articles available have indicated the negativity of AI usage on rabbit, and the factors that can influence the AIprocess including the sexual desire behavior. Therefore, the objective of this review is to identify the important factors that directly or indirectly affect AI process success, meanwhileinterfering with and/or relying on AI assessment.

Modeling of behavioral responses for successful selection of easy-to-train rams for semen collection with an artificial vagina.

The aim of this study was to analyze the reproductive behavioral responses in Australian Merino rams, to identify those related to a faster training for semen collection with an artificial vagina. Eight Australian Merino rams, aged 1.5 years and with no prior sexual experience, were randomly selected from an extensively grazed flock. One immobilized ewe with no hormone stimulation was used for rams to sexually interact and mount. The frequencies of approaching, sniffing, flehmen, pushing, pawing with chin resting, and tongue flicking were recorded during eight training and three post-training assessments periods. In addition, the duration of sniffing and flehmen responses, as well as the time from when the ram started to approach the ewe until the mount with ejaculation (completed mount) were recorded. Descriptive, correlation, and modeling analyses were performed. Amongst the rams, four mounted the ewe and ejaculated for the first time during the training phase, and three mounted and ejaculated for the first time after the training phase. The remaining ram mounted the ewe and ejaculated for the first time during the post-training evaluation in the following year. A great variability in the behavior repertoire was observed among rams. The correlation analysis indicated that the completed mount was associated with the behaviors during the approaching response. The expression of the sniffing response decreased between the training phase and post-training evaluation, while the responses of pushing the ewe and tongue flicking ceased to occur. Pawing the side of the ewe with the chin resting on the back of the ewe and flehmen responses, however, continued between the training and post-training phases. This led to a decrease in the time from when the ram started to approach the ewe until the completed mount. It is concluded that the responses of approaching the ewe, pawing the side of the ewe with chin resting on the ewe, and sniffing of the ewe (the latter occurring only during the training phase) are behavioral indicators that could be used for selection of easy-to-train rams for purposes of semen collection with an artificial vagina.

Aspectos microbiológicos do sêmen de bovinos mantidos em central de reprodução animal / Microbiological aspects of sperm bovine maintained in the central specialized in animal reproduction

A produção de sêmen congelado nas centrais de reprodução é uma ferramenta importante para a pecuária brasileira, todavia o processo de coleta do sêmen pode influenciar nas características biológicas do ejaculado. Considerando a coleta para a inseminação artificial um procedimento não estéril, podendo promover vários gêneros de bactérias no ejaculado, o objetivo do trabalho foi avaliar os aspectos microbiológicos de amostras de sêmen bovino, in natura e congelado, assim como os diluidores utilizados na rotina em uma central de produção de sêmen. Para o presente trabalho foram utilizados cinco touros da raça nelore, e, destes, avaliados dois ejaculados de cada animal, sendo coletados no intervalo de duas semanas. As colheitas de sêmen foram realizadas pelo método de vagina artificial. Também foram avaliados dois diluidores, Triladyl® e BotuBov®. Realizou-se o experimento pelo método de semeadura em Ágar TSA por esgotamento, e após 24h, 48h e 72 horas foram realizadas análises para a identificação e contagem de colônias crescidas de UFC/mL. As colônias foram avaliadas por coloração pelo método de Gram e microscopia óptica (100x) e classificadas conforme morfologia. Os dados laboratoriais foram submetidos ao teste de Kolmogorov-Smirnov para a verificação de normalidade ou homocedasticidade. Observou-se o predomínio de Bastonetes Gram Negativos e Gram Positivos ao fim do experimento, bem como a presença de outros microrganismos em menores quantidades, sugerindo contaminação nas amostras. Houve presença de microrganismos tanto no sêmen in natura e congelado, quanto nos diluidores comerciais, mesmo seguindo protocolos microbiológicos para garantir a esterilidade das amostras Os resultados demonstram níveis aceitáveis de microrganismos no sêmen e nos diluidores; entretanto, estabelecer novas diretrizes no controle da qualidade microbiológica tende a ser uma linha de trabalho promissora na área da biotecnologia.

The production of frozen semen in the central specialized in animal reproduction is an important tool for the Brazilian livestock. However the semen collection process can influence the biological characteristics of the ejaculated. Considering the collection for artificial insemination as non-sterile procedure, it can promote several types of bacteria in the ejaculated semen. In this context, the aim of this study was to evaluate the microbiological aspects of bovine semen samples, fresh, frozen and thinners used in routine in a insemination center. For this study, it was used five bulls of the Nelore breed, and evaluated two ejaculated semen from each animal, being collected at two weeks interval. Semen samples were collected by the method of artificial vagina. There were also evaluated two thinners, Triladyl® and BotuBov®. Furthermore, the experiment was conducted by seeding method Agar TSA by exhaustion, and after 24 hours; 48h; and 72 hours analysis was performed for identification and counting of colonies grown in CFU / ml. The colonies were assessed by staining by the Gram method and optical microscopy (100x) and classified as its morphology. The laboratory data were submitted to the Kolmogorov-Smirnov test for normality or check homoscedasticity. There was a predominance of Rods Gram Negative and Gram Positive at the end of the experiment, as well as presence of other microorganisms in smaller amounts, suggesting contamination in the samples. There were microorganisms present in both fresh and frozen semen,and in commercial thinners, even following microbiological protocols to ensure sample sterility. The results demonstrate acceptable microorganismslevelsin semen and thinners; however, establishing new guidelines for microbiological quality control tends to be a promising biotechnology area.

Aspectos microbiológicos do sêmen de bovinos mantidos em central de reprodução animal / Microbiological aspects of sperm bovine maintained in the central specialized in animal reproduction

A produção de sêmen congelado nas centrais de reprodução é uma ferramenta importante para a pecuária brasileira, todavia o processo de coleta do sêmen pode influenciar nas características biológicas do ejaculado. Considerando a coleta para a inseminação artificial um procedimento não estéril, podendo promover vários gêneros de bactérias no ejaculado, o objetivo do trabalho foi avaliar os aspectos microbiológicos de amostras de sêmen bovino, in natura e congelado, assim como os diluidores utilizados na rotina em uma central de produção de sêmen. Para o presente trabalho foram utilizados cinco touros da raça nelore, e, destes, avaliados dois ejaculados de cada animal, sendo coletados no intervalo de duas semanas. As colheitas de sêmen foram realizadas pelo método de vagina artificial. Também foram avaliados dois diluidores, Triladyl® e BotuBov®. Realizou-se o experimento pelo método de semeadura em Ágar TSA por esgotamento, e após 24h, 48h e 72 horas foram realizadas análises para a identificação e contagem de colônias crescidas de UFC/mL. As colônias foram avaliadas por coloração pelo método de Gram e microscopia óptica (100x) e classificadas conforme morfologia. Os dados laboratoriais foram submetidos ao teste de Kolmogorov-Smirnov para a verificação de normalidade ou homocedasticidade. Observou-se o predomínio de Bastonetes Gram Negativos e Gram Positivos ao fim do experimento, bem como a presença de outros microrganismos em menores quantidades, sugerindo contaminação nas amostras. Houve presença de microrganismos tanto no sêmen in natura e congelado, quanto nos diluidores comerciais, mesmo seguindo protocolos microbiológicos para garantir a esterilidade das amostras Os resultados demonstram níveis aceitáveis de microrganismos no sêmen e nos diluidores; entretanto, estabelecer novas diretrizes no controle da qualidade microbiológica tende a ser uma linha de trabalho promissora na área da biotecnologia.(AU)

The production of frozen semen in the central specialized in animal reproduction is an important tool for the Brazilian livestock. However the semen collection process can influence the biological characteristics of the ejaculated. Considering the collection for artificial insemination as non-sterile procedure, it can promote several types of bacteria in the ejaculated semen. In this context, the aim of this study was to evaluate the microbiological aspects of bovine semen samples, fresh, frozen and thinners used in routine in a insemination center. For this study, it was used five bulls of the Nelore breed, and evaluated two ejaculated semen from each animal, being collected at two weeks interval. Semen samples were collected by the method of artificial vagina. There were also evaluated two thinners, Triladyl® and BotuBov®. Furthermore, the experiment was conducted by seeding method Agar TSA by exhaustion, and after 24 hours; 48h; and 72 hours analysis was performed for identification and counting of colonies grown in CFU / ml. The colonies were assessed by staining by the Gram method and optical microscopy (100x) and classified as its morphology. The laboratory data were submitted to the Kolmogorov-Smirnov test for normality or check homoscedasticity. There was a predominance of Rods Gram Negative and Gram Positive at the end of the experiment, as well as presence of other microorganisms in smaller amounts, suggesting contamination in the samples. There were microorganisms present in both fresh and frozen semen,and in commercial thinners, even following microbiological protocols to ensure sample sterility. The results demonstrate acceptable microorganismslevelsin semen and thinners; however, establishing new guidelines for microbiological quality control tends to be a promising biotechnology area.(AU)

Feasibility of semen collection in red-winged tinamou (Rhynchotus rufescens) by manual stimulation and sazonality implications.

This study aimed to report in detail, the technique and challenges of cloacal massage, to collect and evaluate semen from red-winged tinamou (Rhynchotus rufescens) keep in captivity, performed by only one technician. Sixty-four semen collection attempts, from 16 adult males, during breeding season and 16 attempts form these same 16 males in non-breeding season, were performed. Prior to collection, all animals were conditioned to cloacal massage for 6 weeks and the ejaculates were succeed with viable spermatozoa and then, evaluated for feces, urine and mucus contamination, volume, concentration, sperm vigor, motility, morphological defects and acrosome integrity. Semen collection success rate was 63% in breeding season and 2 (5%) samples were discarded by grade 5 contamination. Only 3 ejaculates from 16 tinamou were obtained in non-breeding season. Sperm concentration and acrosome integrity was higher (p = 0.00) in breeding season, and the percentage of total sperm morphological defects, were high in both in breeding and out breeding season. Overall, we concluded that the red-winged tinamou breeding season, is linked to photoperiod (spring and summer), and at this period time, semen can be obtained by cloacal massage collection satisfactorily, allowing its use in reproduction biotechnologies and sperm cryopreservation.

Ejaculation does not contribute to the stress response to electroejaculation in sheep.

Electroejaculation procedures (EEPs) provoke stress; nevertheless, ejaculation produces physiological changes similar as those usually used to measure stress responses. The application of EEP to animals that cannot ejaculate-as ewes-may be useful to discriminate the responses induced by ejaculation from those provoked by EEP. The aim was to determine the stress response to EEP in rams and ewes. The EEPs were applied to 10 rams and 10 ewes during the non-breeding season, and the number of vocalizations, the heart rate, rectal temperature, serum cortisol concentration, biochemical and haematological parameters were measured. Overall, EEP provoked increases in cortisol concentration, glycaemia, rectal temperature and concentration of creatine kinase (all them: p < .0001) as well as relative concentration of granulocytes (p = .003) and absolute granulocyte concentration (p = .0002) in both, rams and ewes. Heart rate, relative concentration of lymphocytes (p = .001), haematocrit (p = .02) and haemoglobin (p = .045) decreased in animals from both genders after EEP. Besides, cortisol (p < .0001), rectal temperature (p = .002) and glycaemia (p = .001) were greater in ewes than rams, and creatine kinase also tended to be greater in ewes than rams (p = .054). On the other hand, the number of animals that vocalized (p = .006), white blood cells (p = .02) and absolute lymphocytes (p = .02) were greater in rams than ewes. The general trends show a similar pattern of stress responses in animals from both genders. Therefore, we concluded that ejaculation does not contribute to the stress response provoked by the EEP. This procedure also provokes muscular damage and probably pain.

Cooling of canine semen contaminated with urine

A contaminação do sêmen canino pode ser causada pela micção durante a ejaculação, ou quando o sêmen flui diretamente para a bexiga, que são problemas comuns durante a eletroejaculação para a criopreservação. Os efeitos que a urina causa ao processo de criopreservação, entretanto, não estão completamente entendidos. O presente estudo determinou a resistência do sêmen a diferentes níveis de contaminação por urina após 24h de resfriamento. As amostras de sêmen obtidas de 10 cães foram incubadas em soluções de Ringer, e em diferentes concentrações de urina (0,3,6,13,25,50 e 100%), a 37°C, por 20 min. As amostras foram centrifugadas e os espermatozoides decantados foram ressuspendidos, e resfriados a 5°C por 24h em diluidor a base de leite. A qualidade das amostras de sêmen contaminado por urina diferiu do controle apenas após a exposição de soluções com concentração superior a 13% de urina. Após o resfriamento, apenas os espermatozoides incubados com soluções de urina superiores a 25% exibiram menor atividade que as incubadas na solução simples de Ringer. Em conclusão, o sêmen canino resiste à contaminação com até 13% de urina durante 20 minutos e a motilidade das amostras contaminadas com até 25% recuperam-se de forma semelhante às amostras não contaminadas, se resfriadas a 5°C durante 24 h em um diluidor à base de leite.

Contamination of canine semen with urine, caused by urination during ejaculation or semenflowinto theurinary bladder,is a common problem of sperm collected by electroejaculation for cryopreservation. The effects of urine on sperm cryopreservation, however, are not fully understood. The present study determined the acceptable upper level of contamination with urine for canine semen preservation after 24-h cooling. Semen samples obtained from 10 dogs were incubated with Ringer's solution and different urine concentrations (0, 3, 6, 13, 25 50% and 100%) at 37°C for 20 min. The samples were centrifuged and the decanted sperm resuspended and cooled to 5°C for up to 24 h in a milk-based semen extender. The quality of the contaminated semen samples only differed from that of the control treatment after exposure to urine concentrations above 13%. After cooling, only sperm incubated with urine solutions above 25% exhibited lower activity than those incubated in simple Ringer's solution. In conclusion, canine semen resists contamination with up to 13% urine for 20 min, and the motility of samples contaminated with up to 25% recovers similarly to that of uncontaminated samples if cooled at 5°C for 24 h in a milk-based extender.

Coleta de sêmen para inseminação artificial em cães com desvio ventral do pênis / Semen collection for artificial insemination in dogs with ventral deviation of penis

São apresentados dois casos de desvio peniano ventral em cães. No Caso 1, um Poodle de quatro anosde idade com adequada libido, mas incapacidade de cópula devido a desvio ventral do pênis, cuja coleta desêmen foi realizada e inseminadas artificialmente quatro cadelas. No Caso 2, um Dachshund de um ano e meiode idade com adequada libido, mas desvio ventral do pênis, submetido à coleta de sêmen para espermiograma.Este cão foi capaz de copular uma mesma cadela em dois estros distintos. Ao exame radiográfico do cão do Caso2, foi identificada inflexão ventral do osso peniano de 30º, sem sinais agudos ou crônicos de fratura. Em nenhumdos casos havia outras alterações genitais e urinárias concomitantes. Conclui-se pela variável capacidade decópula e possibilidade de coleta de sêmen para inseminação artificial nestes cães.(AU)

Two cases of ventral penile deviation in dogs are presented. In Case 1, a four years old Poodle withadequate libido but disability of copulation because of ventral penile deviation, whose semen collection wassuccessfully achieved and four bitches artificially inseminated. In Case 2, a one and a half years old Dachshundwith adequate libido but ventral penile deviation, who had semen collected for spermogram. This dog was ableto copulate one bitch in two different heats. At the radiographic examination of the Case 2 dog, it was identifieda ventral penile bone inflection of 30º, without acute or chronic signs of fracture. In none of the cases otherconcomitant genital and urinary abnormalities was observed. The results showed the variable copulationcapacity and the possibility of semen collection for artificial insemination of these dogs.(AU)

Coleta de sêmen para inseminação artificial em cães com desvio ventral do pênis / Semen collection for artificial insemination in dogs with ventral deviation of penis

São apresentados dois casos de desvio peniano ventral em cães. No Caso 1, um Poodle de quatro anosde idade com adequada libido, mas incapacidade de cópula devido a desvio ventral do pênis, cuja coleta desêmen foi realizada e inseminadas artificialmente quatro cadelas. No Caso 2, um Dachshund de um ano e meiode idade com adequada libido, mas desvio ventral do pênis, submetido à coleta de sêmen para espermiograma.Este cão foi capaz de copular uma mesma cadela em dois estros distintos. Ao exame radiográfico do cão do Caso2, foi identificada inflexão ventral do osso peniano de 30º, sem sinais agudos ou crônicos de fratura. Em nenhumdos casos havia outras alterações genitais e urinárias concomitantes. Conclui-se pela variável capacidade decópula e possibilidade de coleta de sêmen para inseminação artificial nestes cães.

Two cases of ventral penile deviation in dogs are presented. In Case 1, a four years old Poodle withadequate libido but disability of copulation because of ventral penile deviation, whose semen collection wassuccessfully achieved and four bitches artificially inseminated. In Case 2, a one and a half years old Dachshundwith adequate libido but ventral penile deviation, who had semen collected for spermogram. This dog was ableto copulate one bitch in two different heats. At the radiographic examination of the Case 2 dog, it was identifieda ventral penile bone inflection of 30º, without acute or chronic signs of fracture. In none of the cases otherconcomitant genital and urinary abnormalities was observed. The results showed the variable copulationcapacity and the possibility of semen collection for artificial insemination of these dogs.

Risk factors for bacterial contamination during boar semen collection.

The aim of this study was to evaluate the influence of multiple factors on bacterial contamination in 213 ejaculates from four boar studs. Semen contamination by aerobic mesophiles increased in ejaculates where the preputial fluid flowed into the collection container, collection glove was dirty, preputial hair was long (>1.0 cm), the collection lasted >7 min and boars were older than 18 months. An increase in coliforms occurred when preputial fluid dripped into the collection container, collections lasted >7 min or when penis escaped during collection. Semen contamination increased when two or more factors related to hygiene (poor hygiene of the boar, dirty preputial ostium, large preputial diverticulum, long preputial hair, dirty gloves, preputial liquid trickling from the hand of the technician into the semen container and penis escaping) were present. A vigilant protocol of collection must be followed to minimize bacterial contamination, especially avoiding dripping of preputial liquid into the semen container.

Utilização de cones de polietileno para coleta de sêmen canino / The use of polyethylene cones for canine semen collection

A utilização de um cone de polietileno produzido com luva de palpação retal para grandes animais (SuperFlex®, WalMur, Argentina) acoplado a tubo de centrífuga para coleta de sêmen foi avaliada em 230 cães de 48 diferentes raças. Houve 94,5% de êxito nas coletas sem traumatismo peniano ou perda de sêmen. Para testar a qualidade do sêmen obtido, ejaculados de 10 cães Pastores Alemães foram submetidos a três tratamentos: 1) controle (C); 2) efusão da amostra pela superfície de um fragmento de polietileno (PP); e 3) manutenção de um fragmento de polietileno mergulhado na amostra (CP). Após 10 min de incubação a 37ºC, foram mensuradas a motilidade progressiva (M) e o vigor espermático (V) das amostras. Houve redução significativa de M e V após incubação nos três grupos em relação à amostra inicial, mas M e V foram semelhantes entre os grupos após incubação. O contato do sêmen com o polietileno por curto período (PP) ou por 10 min (CP) não causou efeito deletério sobre a qualidade seminal. Conclui-se ser viável a utilização desse método de coleta de sêmen canino.(AU)

The use of a polyethylene cone manufactured with large animal rectal examination glove (SuperFlex®, WalMur, Argentina) attached to a centrifuge tube for collection of semen was evaluated in 230 dogs from 48 different breeds. There was a 94.5% success on collection without penile trauma or semen loss. In order to test the quality of semen collected, ejaculates from 10 German Shepherd dogs were subjected to three treatments: 1) control (C); 2) the sample was flowed through the surface of a polyethylene fragment (PP) and 3) one fragment of polyethylene was kept immersed in the sample (CP). After 10 min of incubation at 37ºC, progressive motility (M) and sperm vigor (V) were measured. There was a significant reduction in M and V on the three groups in relation to the initial sample, but M and V were similar among groups after incubation. The contact of semen with polyethylene for a short period (PP) or for 10 min (CP) did not cause deleterious effects on semen quality. It is concluded that is viable to use semen of dogs collected with the proposed method.(AU)

Utilização de cones de polietileno para coleta de sêmen canino / The use of polyethylene cones for canine semen collection

A utilização de um cone de polietileno produzido com luva de palpação retal para grandes animais (SuperFlex®, WalMur, Argentina) acoplado a tubo de centrífuga para coleta de sêmen foi avaliada em 230 cães de 48 diferentes raças. Houve 94,5% de êxito nas coletas sem traumatismo peniano ou perda de sêmen. Para testar a qualidade do sêmen obtido, ejaculados de 10 cães Pastores Alemães foram submetidos a três tratamentos: 1) controle (C); 2) efusão da amostra pela superfície de um fragmento de polietileno (PP); e 3) manutenção de um fragmento de polietileno mergulhado na amostra (CP). Após 10 min de incubação a 37ºC, foram mensuradas a motilidade progressiva (M) e o vigor espermático (V) das amostras. Houve redução significativa de M e V após incubação nos três grupos em relação à amostra inicial, mas M e V foram semelhantes entre os grupos após incubação. O contato do sêmen com o polietileno por curto período (PP) ou por 10 min (CP) não causou efeito deletério sobre a qualidade seminal. Conclui-se ser viável a utilização desse método de coleta de sêmen canino.

The use of a polyethylene cone manufactured with large animal rectal examination glove (SuperFlex®, WalMur, Argentina) attached to a centrifuge tube for collection of semen was evaluated in 230 dogs from 48 different breeds. There was a 94.5% success on collection without penile trauma or semen loss. In order to test the quality of semen collected, ejaculates from 10 German Shepherd dogs were subjected to three treatments: 1) control (C); 2) the sample was flowed through the surface of a polyethylene fragment (PP) and 3) one fragment of polyethylene was kept immersed in the sample (CP). After 10 min of incubation at 37ºC, progressive motility (M) and sperm vigor (V) were measured. There was a significant reduction in M and V on the three groups in relation to the initial sample, but M and V were similar among groups after incubation. The contact of semen with polyethylene for a short period (PP) or for 10 min (CP) did not cause deleterious effects on semen quality. It is concluded that is viable to use semen of dogs collected with the proposed method.