Isolation, proliferation and characterization of endometrial canine stem cells.

In the last decade, progenitor cells isolated from dissociated endometrial tissue have been the subject of many studies in several animal species. Recently, endometrial cells showing characteristics of mesenchymal stem cells (MSC) have been demonstrated in human, pig and cow uterine tissue samples. The aim of this study was the isolation and characterization of stromal cells from the endometrium of healthy bitches, a tissue that after elective surgery is routinely discarded. Multipotent stromal cells could be isolated from all bitches enrolled in the study (n = 7). The multipotency of cells was demonstrated by their capacity to differentiate into adipocytic, osteocytic and chondrocytic lineages. Clonogenicity and cell proliferation ability were also tested. Furthermore, gene expression analysis by RT-PCR was used to compare the expression of a set of genes (CD44, CD29, CD34, CD45, CD90, CD13, CD133, CD73, CD31 CD105, Oct4) with adipose tissue-derived MSC. Stromal cells isolated from uterine endometrium showed similar morphology, ability of subculture and plasticity, and also expressed a panel of genes comparable with adipose tissue-derived MSC. These data suggest that endometrial stromal cells fulfil the basic criteria proposed by the "Mesenchymal and Tissue Stem Cell Committee of the International Society for Cellular Therapy" for the identification of mesenchymal stem cells. Although endometrial mesenchymal stem cells (EnMSC) showed a lower replicative ability in comparison with adipose tissue-derived MSC, they could be considered a cell therapeutic agent alternative to adipose tissue or bone marrow-derived MSC in dog.

Exogenous Sonic hedgehog modulates the pool of GABAergic interneurons during cerebellar development.

All cerebellar GABAergic interneurons were derived from a common pool of precursor cells residing in the embryonic ventricular zone (VZ) and migrating in the prospective white matter (PWM) after birth, where both intrinsic and extrinsic factors contribute to regulate their amplification. Among the environmental factors, we focused on Sonic hedgehog (Shh), a morphogen well known to regulate neural progenitor cell proliferation. We asked if and how exogenous Shh treatment affects the lineage of cerebellar GABAergic interneurons. To address these issues, exogenous Shh was administered to embryonic and postnatal organotypic slices. We found that Shh is able to expand the pool of interneuron progenitors residing in the embryonic epithelium and in the postnatal PWM. In particular, Shh signalling pathway was highly mitogenic at early developmental stages of interneuron production, whereas its effect decreased after the first postnatal week. Gene expression analysis of sorted cells and in situ hybridization further showed that immature interneurons express both the Shh receptor patched and the Shh target gene Gli1. Thus, within the interneuron lineage, Shh might exert regulatory functions also in postmitotic cells. On the whole, our data enlighten the role of Shh during cerebellar maturation and further broaden our knowledge on the amplification mechanisms of the interneuron progenitor pool.

Use of contrast-enhanced ultrasonography in chronic pathologic canine testes.

Contrast-enhanced ultrasound with sulphur hexafluoride microbubbles was performed in seven healthy dogs without a history of reproductive pathology and with histologically confirmed normal testes and in 42 dogs with chronic scrotal anomalies. All dogs underwent orchiectomy and histological examination. Enhancement patterns and perfusion parameters (peak intensity and regional blood flow) of testes of healthy dogs and testes with chronic lesions were compared. Fourteen non-pathologic and 60 pathologic testes were considered. Forty testes were neoplastic (24 interstitial cell tumours, 9 seminomas, 7 Sertoli cell tumours), 20 were non-neoplastic (16 testicular degenerations, 2 chronic orchitis, 1 testicular atrophy, 1 interstitial cell hyperplasia). In healthy dogs, the contrast medium flow had a rapid homogeneous wash-in and wash-out, with a short peak phase. With contrast ultrasound, testes that were inhomogeneous with a hyperenhancing pattern were associated with neoplasia (sensitivity: 87.5%, specificity: 100%). Lesions with persistent inner vessels and a hypo-to-isoechoic background were significantly associated with seminomas (sensitivity: 77.8%, specificity: 100%). Testes with non-neoplastic lesions were characterized by a scant/moderate homogeneous enhancement. Perfusion parameters were higher in neoplastic lesions. Contrast ultrasound was a feasible diagnostic tool in the assessment of testicular lesions, with hyperenhancement being an important feature in the diagnosis of malignancy.

A novel blood-sampling technique in lactating sows: The mammary vein route.

In the pig industry, labour efficiency and animal welfare have become two of the most important factors for achieving technical goals and farming competitiveness. Blood sampling is one of the most common sample-collecting techniques, but routine on-field blood collection can be very demanding for farm operators and the welfare of the animals, in particular for lactating sows. The aim of this study was to describe and investigate the mammary vein as a novel means of blood access in lactating sows that does not require coercive restraint. The study involved a total of 68 sows: 34 animals were sampled from the jugular vein (Group J) and the other 34 sows from the mammary vein (Group M). Labour time and indicators of the sow welfare (vocalizations during collection and serum cortisol concentration in the 30min after the procedure) were collected from the two groups. The total amount of labour required, calculated as the time employed to perform blood collection multiplied by the number of operators involved in performing the technique (one for Group M and two for Group J, one for restraint and one for sampling), was significantly lower in Group M than Group J (Group M, 39.83±29.45s; Group J, 82.73±55.34s; P<0.001). Mean blood volume collected at T0 was 4.81±2.00mL and 4.84±1.73mL in Group J and Group M, respectively (P>0.05). The percentage of sows that vocalized in Group M was less than in Group J (2.94% vs. 94.12%; P<0.001). Serum cortisol concentrations were not statistically different between the two groups (P>0.05). The present study suggests greater efficiency in terms of saving labour time and reducing sow vocalization during blood collection with the use of mammary vein access compared with jugular vein access.

Early diagnosis of pregnancy in sows by ultrasound evaluation of embryo development and uterine echotexture.

Two operators attempted to detect pregnancy ultrasonographically in 196 sows daily from 15 to 25 days after insemination; 20 unbred sows were also investigated. The probe was applied transcutaneously on the right abdominal wall near the last three mammary glands. During each examination, the embryos were visualised and their transverse and longitudinal dimensions were measured. Pregnancy was confirmed by an ultrasonographic detection of embryos five days after the first ultrasound diagnosis and finally 30 to 32 days after insemination. The accuracy of diagnosis was less than 83 per cent on days 15, 16 and 17 but improved to more than 90 per cent from day 18 onwards. The uterine echotexture was studied in seven sows at oestrus and 15, 17, 19, 21, 23 and 25 days after insemination. The echotexture was more homogeneous from days 15 to 25 after insemination than at oestrus.

Complete uterine prolapse without uterine mucosal eversion in a queen.

A five-year-old female cat weighing 3 kg was presented by the owner after noticing a large pink, bilobed mass protruding through the vulva during labour. The cat was in good condition, with appropriate lactation, and the newborn kittens were nursing normally. The uterus was not reverted or invaginated at examination, and there was rupture of the mesovarium, mesometrium and uterine-vaginal connection around the cervix. Manual reduction of the prolapsed uterus was not possible because of torn ligaments. A coeliotomy was performed to remove the ovaries, and the apex of the uterine horns was passed by the vaginal route. The remaining part of the mesometrium was disconnected, and the prolapsed uterus was removed. The queen and kittens were discharged from the hospital on the second day after surgery. An unusual feature of this case is that the prolapse was complete, without eversion of any part of the uterus through a vaginal tear.

Relationship between body fat and body condition score and their effects on estrous cycles of the Standardbred maiden mare.

Standardbred maiden mares are generally stressed and in poor physical condition because of their incorrect management at the end of their racing careers. The purpose of this study was to identify an objective assessment that, similar to body condition score (BCS) determination, is easy to measure and able to confirm or improve fattening status assessment, as well as to ascertain whether a relationship with reproduction efficiency exists in subjects destined for a first-time insemination program. The authors assessed 29 Standardbred maiden mares (7 +/- 2 years old) during the breeding season. On January 15 (day 0), the same operator performed the following on all subjects: the first gynecological and ultrasound examination, a BCS assessment (range 0 to 5), and an adiposity objective assessment (i.e., measurement of fat thickness by ultrasound scan). At day 0, all mares were in seasonal anestrous. On ovulation day, all subjects exited the study. Both techniques were shown to be significantly and reciprocally correlated (r = 0.976; P < 0.01) to the first seasonal ovulation in maiden mares (-0.772 and -0.805, respectively, for fat thickness and BCS; P < 0.01). Based on the results obtained, regression equations for the prediction of days to the first seasonal ovulation (y) were created. The best predictive equation was the following: y = 26.714x(3) - 202.44x(2) + 446.04x - 195.65 (R(2) = 0.783; SE = 17 d; P0.01), with an independent BCS variable. In conclusion, this study suggests that increasing the plane of nutrition (i.e., flushing), starting approximately 3 weeks before the breeding season, stimulates ovarian activity in stressed maiden mares.

Ultrasonography and cystic hyperplasia-pyometra complex in the bitch.

Cystic endometrial hyperplasia-pyometra complex is the most frequent and important endometrial disorder encountered in bitches. The pathogenesis of the disease is related to the activity of progesterone [Feldman and Nelson, Canine and Feline Endocrinology and Reproduction (1996) W.B. Saunders, Philadelphia]. Cystic endometrial hyperplasia (CEH) is an abnormal response of the bitch's uterus to ovarian hormones [De Bosschere et al. Theriogenology (2001) 55, 1509]. CEH is considered by many authors to be an exaggerated response of the uterus to chronic progestational stimulation during the luteal phase of the oestrous cycle, causing an abnormal accumulation of fluid within the endometrial glands and uterine lumen (De Bosschere et al. 2001). The resulting lesions of pyometra are due to the interaction between bacteria and hormones. The aim of this study was to evaluate if transabdominal uterine ultrasonography can be a useful and reliable diagnostic method to confirm Dow's [Veterinary Record (1958) 70, 1102] and De Bosschere's histopathological classification of CEH-pyometra complex. The study was carried out on 45 bitches with pyometra, 10 purebreds and 35 crossbreeds, 1-15 years old, 20% of which had whelped at least once. None of these animals had received exogenous oestrogen or progesterone treatment. On admission the 45 animals were in the luteal phase of the oestrus cycle. Clinical signs, blood parameters, uterine ultrasonography, bacterial swabs and uterine histopathological results were recorded. Results suggest that ultrasonographic examination is a useful and reliable tool for the diagnosis of cystic endometrial hyperplasia.