Insights into the influence of canine breed on proteomics of the spermatozoa and seminal plasma.

This study aimed to characterize the proteome of spermatozoa and seminal plasma of 4 purebred dogs (Golden Retriever, Great Dane, Bernese Mountain Dog, and Maremmano-Abruzzese Sheepdog). The ejaculate of 13 dogs was collected, and sperm characteristics were subjectively evaluated. Seminal plasma and sperm cells were separated and prepared individually for mass spectrometry. Data were evaluated by univariate and multivariate statistical analysis. A total of 162 proteins were identified, 47 in spermatozoa, 109 in seminal plasma, and 6 in both samples. Serum albumin in spermatozoa and tubulin alpha-3E chain, acrosin binding protein, and tubulin alpha-3 chain in plasma seminal were statistically relevant. Serum albumin and acrosin binding protein improve the sperm capacitation, acrosome reaction, and seminal quality. The tubulin family proteins are related to structural cell organization and flagella movement, and their presence in seminal plasma may be related to sample handling. According to cluster formation, a high association was observed among Bernese Mountain Dog and Great Dane, Golden Retriever, and Maremmano-Abruzzese Sheepdog for sperm proteins. For seminal plasma proteins, Bernese Mountain Dog, Great Dane, and Maremmano-Abruzzese Sheepdog were related. Further studies on breed-specific proteins in the semen of purebred dogs need to be performed to clarify its fertility roles. SIGNIFICANCE: For the first time spermatozoa proteins of dogs are described. The comparison of spermatozoa and seminal plasma proteins of four purebred dogs were performed. These results supporting that differences in semen protein profile of different canine breeds exist, which can improve the biotechnologies of reproduction in this species.

Proteomic data of seminal plasma and spermatozoa of four purebred dogs.

Semen contains several proteins that are important to fertilization and to identify reproductive failures. There are proteins that are specie-specific expressed, although differs among several breeds. This article provides experimental data describing the protein profile of seminal plasma and spermatozoa of four healthy purebred dogs: Golden Retriever (n=3), Bernese Mountain Dog (n=4), Great Dane (n=3), and Maremmano-Abruzzese Sheepdog (n=3), housed at São Paulo state, Brazil. Semen samples were collected by manual stimulation of the penis in a presence of a teaser bitch, when possible. The seminal plasma and sperm cells were separated by centrifugation and prepared for mass spectrometry. The gene ontology annotation of the proteins found is described. This is the first time that proteomic profile of the semen of purebred dogs is described. These data are a valuable resource to improve the biotechnologies of reproduction applied to canid species.

Influence of cooling temperature in sperm subpopulations of domestic cats.

The objective of this study was to identify and compare domestic feline sperm subpopulations, chilled at -1 °C for 24 and 48 h, as well as to analyze the sperm frequency in different subpopulations. Ten adult cats were used. Sperm collection was performed using electroejaculation (EEJ). Spermatic kinetics were evaluated using a computerized system at three moments: fresh, 24 and 48 h after refrigeration. The ejaculates were divided into a group refrigerated at -1 °C (n = 5,) and a group refrigerated at 4 °C (n = 5),. A total of 1560 spermatozoa were analyzed individually, and the sperm subpopulations were identified using multivariate statistics. Three spermatic subpopulations were defined using prior analysis of the hierarchical dendrogram. A principal components analysis (PCA) identified the existence of three groups with higher iterations at the three moments: PC1 (VAP, VCL, VSL, ALH, SVI), PC2 (STR, LIN, WOB and SMI) and PC3 (BCF). Subpopulation 1, after 48 h of refrigeration at -1 °C, and subpopulation 3, after 24 h of refrigeration at 4 °C, maintained their sperm quality, which allowed us to characterize the groups of spermatozoa that were resistant to cryopreservation. The present study identified three well defined ejaculate spermatozoa subpopulations, with proportional distributions between the groups and two refrigeration resistant subpopulations.

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.

Effects of chemical castration using 20% CaCl2 with 0.5% DMSO in tomcats: Evaluation of inflammatory reaction by infrared thermography and effectiveness of treatment.

This study used infrared thermography to monitor the immediate inflammatory reaction to an intratesticular injection of 20% (calcium chloride) CaCl2 with 0.5% dimethyl sulfoxide (DMSO) for chemical castration and to provide evidence of the treatment's clinical viability. Six animals received a 0.25 mL intratesticular injection of 20% CaCl2 with 0.5% DMSO in each testis. Thermographic imaging, testicular measurement, penile spine evaluation, electroejaculation, and sperm analysis were performed before the injection. Eighty days post-injection, the cats were evaluated again, then received an orchiectomy followed by histological analysis. Infrared thermography of the testicular area was performed before (M0) and after anesthesia (M0A); after electroejaculation (M0E); 10 min (M1), 1 h, (M2) and 6 h (M3) after the injection for seven consecutive days (M4-10); after 15 (M11) and 30 days (M12); and after 80 days, before and after anesthesia and after electroejaculation (M13, M13A and M13E). No cats had a significant increase in testicular temperature or behavioral changes. All animals were azoospermic and had a 50% reduction in total testicular volume after 80 days. Histologically, the testes showed different degrees of degeneration, necrosis, calcification and replaced connective tissue, as well as Leydig cell hyperplasia in 7/12 of the testes. Infrared thermography efficiently diagnosed and monitored scrotal inflammation caused by intratesticular injection of 20% CaCl2 solution with 0.5% DMSO, and it is suggested that a single injection of this compound can cause azoospermia after 80 days. We concluded by infrared thermography with a clinical exam, that the adverse reactions were minimal and did not interfere with animal welfare.

Aspectos emocionais gerados pela morte do animal de estimação / Emotional aspects generated by the death of a pet / Aspectos emocionales generados por la muerte de una mascota

A ligação entre o homem e os animais de companhia tem sido cada vez mais intensa, assim como os benefícios dela resultantes. As emoções identificadas no proprietário que perde o seu animal de estimação evidenciam a importância deste no núcleo familiar. Por meio da aplicação de um questionário, verificou-se que os sentimentos mais comuns gerados pela perda do animal de estimação foram: tristeza, nó na garganta, choro e sensação de falha. É importante que o veterinário identifique esses sentimentos, respeite e seja solidário, contribuindo para a evolução e resolução das etapas que compreendem o processo do luto.

The connection between a man and a fellow animal has been increasingly intense as well as its benefits. The emotions identified in an owner who loses his pet highlight the importance of the family. Through the application of a questionnaire, it was found that the most common feelings generated by the loss of a pet were: sadness, knot in the throat, crying and feeling of failure. It is important that the veterinarian identifies such feelings, respects and has solidarity to contribute to the evolution and resolution of the steps that comprise the process of mourning.

La conexión entre el hombre y los animales de compañía ha sido cada vez más intensa, así como las ventajas de ella resultantes. Las emociones identificadas en el propietario que pierde a su mascota evidencian la importancia de este en el núcleo familiar. A través de la aplicación de un cuestionario, se constató que los sentimientos más comunes generados por la pérdida de los animales de compañía fueron: tristeza, nudo en la garganta, llanto y sentimiento de fracaso. Es importante que el veterinario identifique esos sentimientos, respete y sea solidario, contribuyendo para la evolución y resolución de las etapas que comprenden el proceso de luto.