Cytogenetic and molecular insight into the genetic background of disorders of sex development in seventeen cats.

The genetic background of feline disorders of sex development (DSDs) is poorly understood. We performed comprehensive cytogenetic, molecular, and histological studies of 17 cats with abnormal external genitalia, unusual behavior, or tricolor coats (atypical in males). The DSD phenotype of three cats was associated with sex chromosome abnormalities: X/Y translocation (38,XXSRY+), 37,X/38,XY mosaicism, and XX/XY leukocyte chimerism. The remaining 14 affected cats were classified as XY DSD (SRY-positive). In this group and 38 normal males, we analyzed a priori selected candidate genes (SRY, TAC3, CYP11B1 and LHCGR). Only a previously reported nonpathogenic variant was found in SRY. Moreover, SRY gene copy number was determined, and three variants were observed: 6, 5 (modal), and 4 copies in a single DSD case. The known variants in TAC3 and CYP11B1, responsible for testicular hypoplasia, persistent primary dentition or congenital adrenal hyperplasia, were not found in the study group. Nine novel polymorphisms were identified in the LHCGR gene, one of which, a potentially regulatory indel variant in 5'UTR, was significantly associated (p = 0.0467) with XY DSD. Our report confirmed that abnormalities of sex chromosomes are important causes of feline DSDs. We also showed that the indel variant of LHCGR can be considered a promising marker associated with XY DSD phenotype.

Mineral status and enteric methane production in dairy cows during different stages of lactation.

BACKGROUND: Lactating dairy cows are the greatest livestock contributor of methane, a major global greenhouse gas (GHG). However, good feeding management with adequate mineral intake can offers an effective approach to maintaining high levels of milk production and the health of dairy cows over the entire course of lactation, while also helping to reduce methane emission. The study described here investigated the plasma concentrations of both macroelements (Ca, Na, K, Mg, P) and microelements (Zn, Cu, Fe, Mn), as well as enteric methane emission and milk composition in high-yielding dairy cows in different lactation periods. The experiment was performed on Holstein-Friesian dairy cows with the average milk yield of 41 (± 9) L/day in a Polish commercial farm with modern dairy systems. A total of thirty high-yielding dairy cows were randomly assigned into three groups differing by lactation stage: early stage (Early, days 25-100), middle stage (Middle, days 101-250), and late stage (Late, day 250 and later). Dietary treatment for all cows was a total mixture ration (TMR) with maize and alfalfa silage the main forage components. RESULTS: The greatest milk yield and methane production were recorded in early-stage lactating cows, but the greatest methane intensity per kg of corrected milk was recorded in the late stage of lactation. Plasma concentrations of macroelements and microelements did not differ by lactation stages, but increased plasma concentrations of Zn and Fe and decreased plasma levels of Mg were noted during lactation. A positive correlation was found between plasma levels of Mg and other macroelements (Ca, Na, K), and between the concentrations of Fe and Zn, P in plasma, but no correlation between methane emission and mineral status was detected in the different lactation stages. CONCLUSIONS: Our results showed different mineral requirements and enteric methane emissions in each lactation stage. The feeding strategy and mineral utilization were adequate to maintain the health, mineral status, and milk production of the Holstein cows during the entire lactation period, and suggest an effective way of reducing methane emission.

Changes in Leukogram and Erythrogram Results in Bitches with Vaginitis.

Vaginitis in female dogs is a problem most veterinarians face in their practice. It manifests as localized inflammation, and its variable etiology and different severities often make diagnosis problematic. The study consisted of comparing blood smears taken from 16 animals: 8 healthy bitches and 8 bitches with confirmed vaginitis. We analyzed the percentage of different types of white blood cells (leukogram) and changes in the shape of red blood cells (erythrogram) in both groups. We observed changes in red blood cell morphology, i.e., a higher percentage of lacrimocytes and schistocytes in female dogs with vaginitis compared to their healthy counterparts. The observed hematological changes may illustrate the severity of inflammation. The analysis of erythrograms showed a significantly higher percentage of lacrimocytes and schistocytes in diseased bitches (1.58 ± 1.19% and 0.13 ± 0.12%) compared to healthy animals (0.58 ± 0.38 and 0.00 ± 0.00, respectively). The obtained results may indicate that the analysis of erythrograms throughout the course of vaginitis in bitches may constitute a diagnostic tool, as opposed to the analysis of leukograms, which is more sensitive when it comes to the systemic inflammatory response of the organism. It seems that simultaneous analysis of erythrograms and leukograms may facilitate the diagnostic process in clinical practice.

Non-Invasive Measurement of Thyroid Hormones in Domestic Rabbits.

Thyroid hormones are essential for metabolism, energy homeostasis and reproduction. Hormones can be measured in various biological source materials: blood, feces, urine, saliva and others. The aim of our study was to verify usefulness of thyroid hormone analysis in the urine and feces of the domestic rabbit (Oryctolagus cuniculus f. domesticus), comparing them with the serum analyses. Samples were collected from 27 does in the age of 12-14 weeks. Total thyroxine (tT4), total triiodothyronine (tT3), free thyroxine (fT4) and free triiodothyronine (fT3) were tested using the radioimmunological method in serum, feces and urine. The highest concentration of tT4 was found in feces (104.72 ± 59.52 nmol/mg) and the lowest in urine (3.03 ± 3.11 nmol/mL). The highest tT3 concentration was found in blood serum (3.19 ± 0.64 nmol/L) and the lowest in urine (0.31 ± 0.43 nmol/L). The highest concentration of fT4 was observed in feces (43.71 ± 4.79 pmol/mg) and the lowest in blood serum (14.97 ± 3.42 pmol/L). The statistically highest concentration of fT3 (28.56 ± 20.79 pmol/L) was found in urine, whereas the lowest concentration of this hormone was found in feces (3.27 ± 1.33 pmol/mg). There was a positive and statistically significant correlation between serum and urine fT3 (r = 0.76) and a high positive correlation between serum and feces fT3 concentration (r = 0.62). Correlations between concentrations of other thyroid hormones between serum, urine and feces were found to be insignificant. The results suggest that fT3 can be accurately and reliably measured in the feces and urine of the domestic rabbit.

The vaginal microflora changes in various stages of the estrous cycle of healthy female dogs and the ones with genital tract infections.

BACKGROUND: Inflammatory diseases of reproductive tract in bitches are a common problem in veterinary practice. The inflammation can lead to serious health problems. Research to determine the correlation between the health status of females, phase of the cycle, age and bacterial flora of the genital tract has been ongoing for years, but the results obtained by individual authors are often contradictory. RESULTS: A total of 39 dogs were included in this study. Ten were qualified to the 1st group with genital tract infections (8 in anestrus and 2 in proestrus) and 29 to the 2nd group without such infections (16 in anestrus, 9 in proestrus and 4 in diestrus). The most common bacterial isolates obtained from the vaginal tract of all dogs were Escherichia coli, Staphylococcus pseudintermedius and Streptococcus canis. The prevalence of Gram-negative rods (other than E. coli) was significantly higher in the group with genital tract infections versus healthy dogs. There was no presence of Chlamydiaceae, Chlamydia abortus and lactic acid-producing bacteria in tested swabs. CONCLUSIONS: Our study identified the most common bacteria in the genital tract of bitches. The total number of bacteria was almost the same in the healthy and infected dogs, as well as between the cycle stages. In our opinion, bacterial culturing of vaginal swab specimens from bitches without signs of genital disease is of little value. Furthermore, it should always be preceded by clinical examination and cytological examination of the vaginal epithelium.

The domestic cat as a research model in the assisted reproduction procedures of wild felids / Kot domowy jako model badawczy w procedurach wspomaganego rozrodu dzikich kotowatych.

The assisted reproduction techniques are an essential part of endangered species conservation programs. All wild felids are threatened with extinction, and the domestic cat is a good and easily available model for research on wild felids. The procedures used in cats and wild felids include artificial insemination, oocytes collection from female and sperm collection from male, in vitro maturation of oocytes, in vitro fertilization by IVF and ICSI, embryo culture, embryo transfer, cryopreservation of gametes and embryos. The effectiveness of individual procedures in cats and wild felids is often much lower than in humans, livestock or laboratory animals, but the success achieved so far confirms the need for further research.

Myocardial proteomic profile in pulmonary arterial hypertension.

Pulmonary arterial hypertension (PAH) is a rare, fatal, and incurable disorder. Although advances in the understanding of the PAH pathobiology have been seen in recent years, molecular processes underlying heart remodelling over the course of PAH are still insufficiently understood. Therefore, the aim of this study was to investigate myocardial proteomic profile of rats at different stages of monocrotaline-induced PAH. Samples of left and right ventricle (LV and RV) free wall collected from 32 Wistar rats were subjected to proteomic analysis using an isobaric tag for relative quantitation method. Hemodynamic parameters indicated development of mild elevation of pulmonary artery pressure in the early PAH group (27.00 ± 4.93 mmHg) and severe elevation in the end-stage PAH group (50.50 ± 11.56 mmHg). In early PAH LV myocardium proteins that may be linked to an increase in inflammatory response, apoptosis, glycolytic process and decrease in myocardial structural proteins were differentially expressed compared to controls. During end-stage PAH an increase in proteins associated with apoptosis, fibrosis and cardiomyocyte Ca2+ currents as well as decrease in myocardial structural proteins were observed in LV. In RV during early PAH, especially proteins associated with myocardial structural components and fatty acid beta-oxidation pathway were upregulated. During end-stage PAH significant changes in RV proteins abundance related to the increased myocardial structural components, intensified fibrosis and glycolytic processes as well as decreased proteins related to cardiomyocyte Ca2+ currents were observed. At both PAH stages changes in RV proteins linked to apoptosis inhibition were observed. In conclusion, we identified changes of the levels of several proteins and thus of the metabolic pathways linked to the early and late remodelling of the left and right ventricle over the course of monocrotaline-induced PAH to delineate potential therapeutic targets for the treatment of this severe disease.

Meiotic Status Does Not Affect the Vitrification Effectiveness of Domestic Cat Oocytes.

Cryopreservation is important for animal fertility and biodiversity. Unfortunately, cryopreservation of feline oocytes is still an experimental technique. The aims of this study were to analyze the potential toxicity of the cryoprotectants in the vitrification solution (VS) on cat oocytes and to investigate whether the meiotic status of oocytes influences their developmental potential after vitrification. Two experiments were conducted with the VS composed of 20% ethylene glycol, 20% dimethyl sulfoxide, 20% fetal calf serum, 1.5 M trehalose, and 10% Ficoll PM-70: (1) toxicity assessment of the VS on immature cumulus oocyte complexes (COCs), and subsequently in vitro maturation (IVM) and in vitro fertilization; (2) assessment of the influence of the meiotic status on vitrification effectiveness, where immature and in vitro matured COCs were vitrified on the Cryotop. After rewarming, vitrified oocytes were subjected to IVM (immature) and intracytoplasmic sperm injection (ICSI) with fresh epididymal sperm. The toxicity test revealed no negative effect of oocyte exposure to the applied VS on their developmental potential (p > 0.05). Although the vitrification procedure itself significantly reduced the meiotic competence of oocytes, their meiotic status before vitrification (immature vs. in vitro matured) did not influence fertilization and morula rates. The only parameter affected by vitrification was the rate of oocytes suitable for ICSI, which was significantly lower for immature oocytes. Regardless of the meiotic status of vitrified oocytes, morphologically normal morulae were obtained. Moreover, the two meiotic stages examined are suitable for vitrification, with mature oocytes being a better choice when a well-equipped laboratory is available.

Changes in heart morphometric parameters over the course of a monocrotaline-induced pulmonary arterial hypertension rat model.

BACKGROUND: Aim of this study was to assess changes in cardiac morphometric parameters at different stages of pulmonary arterial hypertension (PAH) using a monocrotaline-induced rat model. METHODS: Four groups were distinguished: I-control, non-PAH (n = 18); II-early PAH (n = 12); III-end-stage PAH (n = 23); and IV-end-stage PAH with myocarditis (n = 7). RESULTS: Performed over the course of PAH in vivo echocardiography showed significant thickening of the right ventricle free wall (end-diastolic dimension), tricuspid annular plane systolic excursion reduction and decrease in pulmonary artery acceleration time normalized to cycle length. No differences in end-diastolic left ventricle free wall thickness measured in echocardiography was observed between groups. Significant increase of right ventricle and decrease of left ventricle systolic pressure was observed over the development of PAH. Thickening and weight increase (241.2% increase) of the right ventricle free wall and significant dilatation of the right ventricle was observed over the course of PAH (p < 0.001). Reduction in the left ventricle free wall thickness was also observed in end-stage PAH (p < 0.001). Significant trend in the left ventricle free wall weight decrease was observed over the course of PAH (p < 0.001, 24.3% reduction). Calculated right/left ventricle free wall weight ratio gradually increased over PAH stages (p < 0.001). The reduction of left ventricle diameter was observed in rats with end-stage PAH both with and without myocarditis (p < 0.001). CONCLUSIONS: PAH leads to multidimensional changes in morphometric cardiac parameters. Right ventricle morphological and functional failure develop gradually from early stage of PAH, while left ventricle changes develop at the end stages of PAH.