Ada Rota

Hypoosmotic swelling (HOS) as a screening assay for testing in vitro fertility of bovine spermatozoa

Abstract Semen from 5 Piedmontese bulls was subjected to the hypoosmotic swelling (HOS) test in order to determine if the results could be correlated to the fertilizing capacity in vitro. Semen was routinely prepared for in vitro fertilization (IVF), with aliquots being sampled after thawing, after separation on a Percoll gradient and after capacitation in a medium containing heparin. The aliquots were added to a fructose-sodium citrate hypoosmotic solution (100 mOsm) and incubated at 37 °C for 5 min. At least 200 spermatozoa were observed at x 400 and classified according to the presence or the absence of a swollen tail. After capacitation, spermatozoa were used to fertilize in vitro-matured bovine oocytes (1.5 × 106 cells/mL); IVF was performed in Fert-TALP medium supplemented with 6 mg/mL BSA and 10 μL/mL heparin in a humidified atmosphere with 5% CO2 and 5% oxygen. Presumptive zygotes were cultured in SOF medium supplemented with 8 mg/mL BSA and amino acids. There were no significant differences in the in vitro fertility of the bulls, but a significant difference was found between bulls in the response to the HOS test. The 3 assays were significantly correlated, while no significant correlation was observed between the percentage of swollen spermatozoa and in vitro fertility. The HOS test does not appear to be sufficiently sensitive to discriminate between semen samples of intermediate fertility like those used in this preliminary research.

Age dependent changes in plasma anti-Müllerian hormone concentrations in the bovine male, female, and freemartin from birth to puberty: relationship between testosterone production and influence on sex differentiation.

To understand the behaviour of the gonads, in terms of hormonal secretion, in a model of intersexual development naturally occurring in mammals, we determined plasma concentrations of testosterone, progesterone, and anti-Müllerian hormone (AMH) in bovine freemartins, and compared them to normal levels measured in males and females from birth to puberty. We found that newborn males and freemartins have very high concentrations of AMH (over 700ng/ml). Conversely, plasma AMH concentration is always below 120ng/ml in females. While values remain stable in males for the first five months of life, they sharply decrease in the freemartins within the first fortnight, and reach female levels, which demonstrates that AMH is essentially originated in the male twin. In young bulls the trend of plasma testosterone concentrations is opposite to that of the AMH. The rise in testosterone production at puberty corresponds to a sharp decline in AMH concentrations. Bovine plasma concentrations of AMH are surprisingly higher than those measured in other mammals, including man and mouse. The results obtained are discussed in reference to comparative aspects of endocrine functions.

Aging and Veterinary Care of Cats, Dogs, and Horses through the Records of Three University Veterinary Hospitals

The present article examines over 63,000 medical records belonging to the Veterinary Hospitals of the Universities of Bologna, Torino, and Padova, all in Northern Italy, and relative to dogs (approximately 50,000), cats (approximately 12,000), and companion horses (slightly less than 1,000). The animals of the three species were divided into age classes and categorized per sex into males, females, and neutered individuals. The mean age at visit and the effects of age classes and category (analyzed via ANOVA) are presented and discussed. The data indicate that many animals are presented to the hospitals either in the early phases of their life (presumably for vaccination and, in cats and dogs, gonadectomy) or in the advanced age (over 10u2009years in dogs, over 15u2009years in cats, and over 17u2009years in horses). The records of very old individuals of the three species are also reported. On the whole, the results suggest that a growing population of mature to old domestic carnivores or companion horses reaches ages that were considered exceptional only a few years ago. The data also testify an evolution in the animal–owner relationship and a renewed respect for the value of life in companion domestic mammals.

Detection of Helicobacter in Gastric Washing of Cats

The aim of the current study was to evaluate the efficacy of gastric washing for detecting the presence of Helicobacter spp. in feline gastric mucus. Gastric fluids were collected from 11 cats undergoing routine surgical procedures. The fluids were centrifuged and the pellets were subjected to cytological examination by May-Grünwald/Giemsa (MGG) staining and testing by polymerase chain reaction (PCR) for the presence of Helicobacter after DNA extraction. Helicobacter spp. were detected in 8 of 11 samples by MGG staining and in 10 of 11 samples by PCR. Gastric washing proved to be a useful sampling technique and a valuable alternative to the gold-standard method of gastric biopsy, which has the risk of missing colonized spots in the case of patchy Helicobacter colonization. Gastric washing, a noninvasive way of sampling, could be applied to cats undergoing general anesthesia for different causes. Detection of Helicobacter in cats could allow epidemiological investigations, and sequencing of samples could assist in assessing the distribution of various Helicobacter species.

Barrett’s Disease and Helicobacter Spp. Detection in Dogs with Gastritis

Barrett’s disease (BE) is a metaplastic disorder of the esophageal mucosa in which specialized columnar epithelium replaces healthy squamous epithelium; it is the most common cause of esophageal carcinoma in humans. The dog has been used as a model of Barrett’s esophagus, although the pathology is very rare in the species. In man, infection with Helicobacter pylori resulted inversely associated with BE. The role of Helicobacter spp. in dogs gastritis has not been fully elucidated, as in esophageal disease. Aim of this work was to evaluate esophageal and gastric biopsies of symptomatic dogs in association with the isolation of Helicobacter spp. and to describe and discuss a case of BE.