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Characterization of reproductive traits in Mirandesa beef cattle is important for breed improvement and conservation, mainly due to its little genetic diversity. Reduced individual and maternal performance is often associated with inbreeding depression, which could be further aggravate the environmental effects. In this study, 7386 herd records for Mirandesa were used to characterize the main reproductive traits, like age at first calving (AFC), pregnancy length (PL), calving interval (CI), yearly calving distribution (CD) and productive lifespan (PLf). The non-genetic effects were tested using non-parametric methods, as the target variables were not normally distributed. The median for AFC in Mirandesa was close to 32 months; AFC was affected by the production system, farm and by the year and season of birth. The mean for PL was 287±8.9 days, being affected by parity and calf gender. The median CI, of 378 days, was only affected by the breeding program, parity, season and year. Calving season was unevenly distributed over the year, showing different patterns after the production system. The mean productive lifespan of Mirandesa was 6.45 years, though 20% of the cows presented a PLf longer than ten years. The main non-genetic effects suggested that farmer´s decision and nutrition may constrain the expression of the reproductive traits in Mirandesa breed. This aspect needs to be addressed when designing any breeding programs which should prioritise for the increase in the number of calves per year along with a careful selection of reproducers to decrease reported inbreeding. The authors wish to thank ACBRM (Associação de Criadores de Bovinos de Raça Mirandesa) for providing access to the Mirandesa cattle Herdbook. info:eu-repo/semantics/publishedVersion

The apparent lack of classical mechanisms for maternal recognition of pregnancy is one of the most intriguing features of canine reproduction. Consequently, similar levels of circulating luteal steroids are observed in pregnant and non-pregnant dogs. However, the early pre-implantation canine embryo locally modulates uterine responses to its presence, facilitating the successful onset of pregnancy. As a part of this interaction, the canine uterus undergoes a species-specific decidualization. Maternal stroma-derived decidual cells develop, the only cells of the canine placenta expressing progesterone receptor (PGR). There exists an acute need for an in vitro stable cell line model for canine decidualization. Therefore, herein our goal was to establish, immortalize and characterize such a cell line. We immortalized three monolayer dog uterine stromal (DUS) cell lines by stably transfecting them with SV40Tag oncogene. Cells retained their mesenchymal character for over 30 passages, as evidenced by VIMENTIN staining. Genomic incorporation of the SV40Tag protein was confirmed by immunofluorescence and Western blot analyses. Cells submitted to a classical in vitro decidualization protocol (N6,2'-O-dibutyryladenosine-3',5'-cyclic monophosphate) revealed upregulated gene levels of selected major decidualization markers (e.g. PRLR, PGR, IGF1, PTGES). Additionally, the basic decidualization capability of PGE2 was demonstrated, revealing increased levels of, for example, PGR and PRLR gene expression, thereby implying its involvement in the progesterone-dependent decidualization in the canine uterus. In summary, our in vitro model with immortalized DUS cell line could serve as an ideal and unique model to study the underlying molecular and endocrine mechanisms of canine decidualization.

In the present study, parameters related to animal performance and carcass quality were evaluated in order to technically characterize crossbreed calves under commercial farm conditions. Sixty-six male calves of Holstein Frisian crossed with Rubia Gallega (n=43; 65.15%), Limousine (n= 16; 24.24%) and Belgian Blue-White (n=7; 10.61%) were used, in a ratio representative for the beef industry of the area. Nutrition and husbandry practices were identical for all the animals. All the calves were slaughtered at the age of 8 months. Results indicated that the Belgian Blue-White crosses were the heaviest, more compact and with the best body conformation, and also a better carcass performance. On the contrary Limousine crosses showed the leanest carcass and the lowest performance. The carcasses of Rubia Gallega crosses showed intermediate characteristics. No statistically significant differences were found in the fatness content between the three genotypes.

Hypospadias is rarely reported in dogs. In this study we present 2 novel cases of this disorder of sexual development and, in addition, a case of hereditary sex reversal in a female with an enlarged clitoris. The first case was a male Moscow watchdog with a normal karyotype (78,XY) and the presence of the SRY gene. In this dog, perineal hypospadias, bilateral inguinal cryptorchidism and testes were observed. The second case, representing the Cocker spaniel breed, had a small penis with a hypospadic orifice of the urethra, bilateral cryptorchidism, testis and a rudimentary gonad inside an ovarian bursa, a normal female karyotype (78,XX) and a lack of the SRY gene. This animal was classified as a compound sex reversal (78,XX, SRY-negative) with the hypospadias syndrome. The third case was a Cocker spaniel female with an enlarged clitoris and internally located ovotestes. Cytogenetic and molecular analyses revealed a normal female karyotype (78,XX) and a lack of the SRY gene, while histology of the gonads showed an ovotesticular structure. This case was classified as a typical hereditary sex reversal syndrome (78,XX, SRY-negative). Molecular studies were focused on coding sequences of the SRY gene (case 1) and 2 candidates for monogenic hypospadias, namely MAMLD1 (mastermind-like domain containing 1) and SRD5A2 (steroid-5-alpha-reductase, alpha polypeptide 2). Sequencing of the entire SRY gene, including 5'- and 3'-flanking regions, did not reveal any mutation. The entire coding sequence of MAMLD1 and SRD5A2 was analyzed in all the intersexes, as well as in 4 phenotypically normal control dogs (3 females and 1 male). In MAMLD1 2 SNPs, including 1 missense substitution in exon 1 (c.128A>G, Asp43Ser), were identified, whereas in SRD5A2 7 polymorphisms, including 1 missense SNP (c.358G>A, Ala120Thr), were found. None of the identified polymorphisms cosegregated with the intersexual phenotype, thus, we cannot confirm that hypospadias may be associated with polymorphism in the coding sequence of the studied genes.

Tumour necrosis factor (TNF), a pleiotropic cytokine that regulates cell growth and differentiation as well as the synthesis of other cytokines, has been identified in the uterus of several species describing a cyclic pattern, eventually under ovarian steroid regulation. Information is yet limited on the presence of TNF protein in the canine endometrium during the oestrous cycle and early pregnancy. This study depicts the temporal immunolocalization of TNF in the bitch endometrium along the oestrous cycle and changes associated with the early steps of embryo invasion. TNF immunolabelling was found in both the stromal fibroblasts and epithelial components of the canine endometrium in all stages studied. Stromal immunostaining was more intense than that of the epithelia, in all the stages of the oestrous cycle. In addition, a tendency for a decrease in the surface epithelium intensity score was found in early dioestrus. A positive glandular content was only observed in anoestrus and proestrus stages. In early pregnancy (days 13-16), TNF immunolabelling was detected at the embryo-maternal surface, in the syncytium cords and the trophoblast, as well in the endometrial stroma and the basal endometrial glands, but not in the lacunar epithelium. The overall TNF immunoreactivity was higher in early pregnancy samples in comparison with those of the early dioestrus and dioestrus stages, suggesting it plays a role during implantation.

Contents Tumour necrosis factor (TNF), a pleiotropic cytokine that regulates cell growth and differentiation as well as the synthesis of other cytokines, has been identified in the uterus of several species describing a cyclic pattern, eventually under ovarian steroid regulation. Information is yet limited on the presence of TNF protein in the canine endometrium during the oestrous cycle and early pregnancy. This study depicts the temporal immunolocalization of TNF in the bitch endometrium along the oestrous cycle and changes associated with the early steps of embryo invasion. TNF immunolabelling was found in both the stromal fibroblasts and epithelial components of the canine endometrium in all stages studied. Stromal immunostaining was more intense than that of the epithelia, in all the stages of the oestrous cycle. In addition, a tendency for a decrease in the surface epithelium intensity score was found in early dioestrus. A positive glandular content was only observed in anoestrus and proestrus stages. In early pregnancy (days 13-16), TNF immunolabelling was detected at the embryo-maternal surface, in the syncytium cords and the trophoblast, as well in the endometrial stroma and the basal endometrial glands, but not in the lacunar epithelium. The overall TNF immunoreactivity was higher in early pregnancy samples in comparison with those of the early dioestrus and dioestrus stages, suggesting it plays a role during implantation. This is the authors’ version of the following article: R Payan-Carreira, M A Pires and Heriberto Rodriguez-Martinez, Tumour Necrosis Factor in the Canine Endometrium: An Immunohistochemical Study, 2011, REPRODUCTION IN DOMESTIC ANIMALS, (46), 3, 410-418. which has been published in final form at: http://dx.doi.org/10.1111/j.1439-0531.2010.01681.x Copyright: Blackwell Publishing Ltd http://eu.wiley.com/WileyCDA/Brand/id-35.html

The authors describe two cases of cervical hypoplasia in jennets, detected during the gynaecological evaluation of two purebred females of the Portuguese donkey breed Asinina de Miranda. These jennets aged 10 and 15 years were referred for consultation by the local breeders association because of infertility detected during an artificial breeding programme. In both jennets the cervix was small, shorter than the usual and closed. In one of the females, outflow of semen during artificial insemination was reported. Clinical and ultrasonographic evaluation of the genital tract demonstrated that both ovaries were normally sized in both animals and presenting structures suggestive of regular ovarian activity. In one of the jennets, a small amount of air in the uterus was evidenced. In the other, signs indicative of atresia of the cervical canal were found. Vaginal examination showed that the cervix was reduced in size and shallowed in both animals. A diagnosis of congenital cervical hypoplasia was made in both cases and the animals were excluded from reproduction.

A 7-year-old intact male Boxer was referred to our services at the Veterinary Teaching Hospital of the University of Trás-os-Montes and Alto Douro, suffering from a persistently erect penis (including the bulbus glandis) that had been exposed for several days. Radiographic and ultrasonographic examinations detected a 5.0 x 3.5 cm mass located dorso-laterally to the urinary bladder. The microbial culture of the mass revealed Staphylococcus spp. At that time, we suspected the involvement of an abscess in the origin of the priapism. Medical and surgical treatments were promptly instituted, which allowed for penile withdrawal into the prepuce; however, the resolution of the penile erection was not accomplished in the following days and penile amputation was required. Histological evaluation of the excised penis revealed extensive infarction of the erectile tissue of the pars longa and bulbus glandis, and also of the blood vessels of the penis. Following penile amputation and antimicrobial therapy, the animal fully recovered. Ultimately, the animal died as a consequence of gastric torsion. At necropsy, some lesions compatible with a previous perforation of the intestinal wall were recorded. The data gathered from the anamnesis, the physical and imaging examinations, along with the post-mortem findings, allowed us to conclude that in this clinical case the primary cause of priapism was a perineal abscess due to bowel perforation.