Sónia Miranda

Temporal changes in neutral endopeptidase/CD10 immunoexpression in the cyclic and early pregnant canine endometrium

CD10 is a multifunctional transmembrane neutral endopeptidase (NEP) that is considered to be a reliable marker of ectopic human endometrial stroma. Available information on NEP/CD10 protein expression in animal endometria is scarce. This study focused on the immunolocalization of NEP/CD10 in the canine uterus and on its temporal changes during the estrous cycle and early pregnancy (Days 11 to 23 post-LH surge) in healthy females. NEP/CD10 expression was found in the canine endometrial stroma in all stages of the estrous cycle, showing cyclic differences both in intensity and in distribution pattern. A small population of negative stromal cells in subsurface position was also observed. This population shared some morphological characteristics with the human predecidual cells, which became positive in progesterone-associated stages of the cycle. In addition, positive immunolabeling was also observed in canine myometrial stroma. In early pregnancy, the basal glandular epithelia and the syncytium cords remained negative to this marker contrasting with the trophoblast and the lacunar epithelium. A weak to moderate intensity of immunolabeling was observed in the decidual cells, whereas stromal immunolabeling was more intense at the delimitation of the syncytium cords. In conclusion, CD10 is consistently expressed in the canine endometrial stroma and myometrium but not in the endometrial epithelia. The characteristic pattern seen in early pregnancy also suggests a role for this molecule in the process of embryo invasion at implantation.

Artificial Insemination in Dogs

In Artificial Insemination (AI) the semen is collected manually from a stud male and thereafter deposited (inseminated) in the female so that fertilization can occur in the absence of natural mating. Artificial Insemination, one of the earliest techniques for assisted reproduction in animals and humans, took longer to be implemented in dogs due to speciesspecific particularities. In past decades, progresses in the knowledge of canine physiology and new advances in canine semen technology allowed these services to become available worldwide. Hence, subsequent to the increase in the artificial insemination demand among dog breeders and owners and the broaden of the AI to preserved semen as a management tool in canine breeding, as through international exchange of frozen semen, inbreeding within breeds can be reduced. Therefore, with spread of canine AI dog, breeders now may select stud dogs from all over the world to improve their kennel genetics, without transport-associated stress to the animals. Also, it is possible to save semen from valuable dogs into sperm bank to be used in next generations, after their death or the peak of reproductive age. In addition, breeders also are aware of the sanitary benefits associated with AI. Avoiding direct contact between the male and female, AI also prevents the spread of sexually transmitted diseases, as those originated by Brucella canis or Herpes virus (Farstad, 2010; Linde Forsberg, 2005a). Although the first reports on AI in dogs subsequent to the Spallanzani experiments (in late XVIII century) appeared by the end of the fifties, reporting the use of fresh semen, or in the sixties, the use of frozen semen, only in the nineties this technique was introduced into dog breeding practice, particularly in USA and Nordic countries (Foote, 2002; England & Millar, 2008). The reproductive physiology of this species and unfavourable response of the dog sperm to freezing were the two major constraints to the initial efforts to improve the AI technique in dogs (Linde Forsberg, 2005a). A lot of research was performed in those areas, especially in the northern Europe, to overcome these issues, generating a large amount of information and allowing technical development, in particular in the canine semen technology. Nowadays, as a consequence of the demand for reproductive technologies, in particular the AI with fresh or refrigerate semen, this is a current service offered in the small animal veterinary practice.

Proliferative Endometrial Lesions Hidden behind the Feline Pyometra

The literature refers to pyometra as the most important pathology in the feline uterus, which is often associated with cystic endometrial disease (cystic endometrial hyperplasia/ pyometra complex or CEH-Pyo). The etiology of pyometra is complex and probably mul‐ tifactorial, but hormonal influences are suggested to play an important role in the patho‐ genesis. Progestagen-based contraceptives may be risk factors for the CEH-Pyo syndrome, for endometrial adenocarcinoma and also to mammary tumors in this species. The histopathological descriptions of pyometra include an enlarged uterus containing purulent fluid, variable endometrial infiltration of neutrophils and bacterial colonization. The degree of hyperplasia of endometrial glands is variable, and frequently the endome‐ trium becomes atrophic. The severity of endometritis is variable. Thereby, the type of in‐ flammatory cells infiltrating the uterine wall or lumen varies accordingly and may include neutrophils, macrophages, plasma cells and lymphocytes. The clinical diagnosis of pyometra is often based on the clinical signs and the physical examination, supported by ultrasound findings. The surgical excision of the uterus is the recommended treatment when the animal is not intent for breeding, as most pyometra clinical signs resolve after ovariohysterectomy. Nevertheless, our clinical practice demonstrated that, in cats, pyometra often masks other uterine conditions that may present a worst prognosis and may interfere with the expect‐ ed outcome. Thus, although seldom requested, the pathological analysis of the uterus with pyometra should be performed following surgery, even if significant macroscopic alterations are not visible, as one frequent finding in pyometra specimens is the co-exis‐ tence of feline endometrial adenocarcinoma (FEA). FEA is usually described as a rare pathology in cats, but recent descriptions suggest that it may be more frequent than thought. Some morphological and clinical features of FEA, as well as molecular markers, have been recently described. Moreover, age is not an ade‐ quate factor for triage, since some FEA cases were described in young animals, prompt‐ ing pathologists, clinicians and researchers into this new reality.

Acute phase proteins and antioxidant responses in queens with pyometra

Acute phase proteins (APP) and biomarkers of oxidative status have proved to be clinically useful biomarkers of pyometra in different species. Despite pyometra is considered one of the most important feline reproductive diseases, information about the APP response and the oxidative status in queens with pyometra is lacking. This study aimed to evaluate the APP and the antioxidant responses at diagnosis and in the post-operative period in feline pyometra. Serum concentrations of serum amyloid A (SAA), haptoglobin (Hp), albumin, total serum thiols (Thiol) and total antioxidant capacity determined by different assays, including trolox equivalent antioxidant capacity (TEAC) assessed by two different methodologies (TEAC1 and TEAC2), ferric reducing ability of plasma (FRAP), and cupric reducing antioxidant capacity (CUPRAC), were determined in 23 queens with pyometra at diagnosis and in 13 healthy control queens submitted to elective ovariohysterectomy. The APP and antioxidants were also evaluated in 11 queens of the pyometra group at days two and 10 after surgery. At diagnosis, queens with pyometra had serum concentrations of SAA, Hp, and FRAP significantly higher (P < 0.001, P < 0.001 and P < 0.05, respectively), and of albumin, Thiol, CUPRAC and TEAC2 significantly lower (P < 0.001, P < 0.001, P < 0.001 and P < 0.01, respectively) than controls. Moreover, concentrations of APP and antioxidants were significantly different (with a tendency to return to physiologic levels) at day 10 after surgery than before surgery. Significant associations were found between APP and antioxidants. According to these results, an APP response and the development of oxidative stress were detected in queens with pyometra. In addition, APP and antioxidants tended to return to physiologic values after surgery in the queens that recovered from the disease. Therefore, our results suggest that APP and selected antioxidants, such as Thiol and CUPRAC, could be potentially useful biomarkers in diagnosis and assessment of the post-operative period in feline pyometra.

In vivo assessment of canine muscle adiposity by real time ultrasonography and image analysis.

Obesity in dogs is a growing concern. Different surveys report a recent increase in dog obesity, their estimates ranging from 22 to over 50%. Surveillance of dog body condition score (BCS) is a routine practice but it is not sensitive enough. Ultrasound (US) assessment of body fat deposits was previously used in different species to predict adiposity, and shows several advantages, like its relative low cost, portability, and great repeatability. In human, intramuscular fat was associated with obesity systemic effects.