Deleterious Effects of Progestagen Treatment in VEGF Expression in Corpora Lutea of Pregnant Ewes

The aim of the current study was to determine the possible effects of progestagen oestrous synchronization on vascular endothelial growth factor (VEGF) expression during sheep luteogenesis and the peri‐implantation period and the relationship with luteal function. At days 9, 11, 13, 15, 17 and 21 of pregnancy, the ovaries from 30 progestagen treated and 30 ewes cycling after cloprostenol injection were evaluated by ultrasonography and, thereafter, collected and processed for immunohistochemical evaluation of VEGF; blood samples were drawn for evaluating plasma progesterone. The progestagen‐treated group showed smaller corpora lutea than cloprostenol‐treated and lower progesterone secretion. The expression of VEGF in the luteal cells increased with time in the cloprostenol group, but not in the progestagen‐treated group, which even showed a decrease between days 11 and 13. In progestagen‐treated sheep, VEGF expression in granulosa‐derived parenchymal lobule capillaries was correlated with the size of the luteal tissue, larger corpora lutea had higher expression, and tended to have a higher progesterone secretion. In conclusion, the current study indicates the existence of deleterious effects from exogenous progestagen treatments on progesterone secretion from induced corpora lutea, which correlate with alterations in the expression of VEGF in the luteal tissue and, this, presumably in the processes of neoangiogenesis and luteogenesis.     

IMAGING DIAGNOSIS—FDG‐PET/CT OF A CANINE SPLENIC PLASMA CELL TUMOR

An 8‐year‐old Shih Tzu developed abdominal pain and hyperglobulinemia. A round splenic mass was noted radiographically and sonographically. The patient was evaluated by fluorodeoxyglucose positron emission tomography coupled with computed tomography (FDG‐PET/CT). There was no evidence of metastasis or bone marrow involvement on PET/CT images. The standardized uptake value (SUV) of the splenic mass was increased over the reference range (SUV=4.83). The patient was diagnosed as splenic extramedullary plasmacytoma through immunohistopathologic study. After the splenectomy, the globulin level normalized and the patient is alive without complications.     

Gross Morphometry of the Bovine Placentome during Gestation

Forty-seven pregnant uteri were collected from an abattoir at eight chronological stages of gestation. Assessments were made of placentome number, weight, length shape and ratio of foetal to maternal tissue by weight. The study showed that there was a great variation in number, size, weight and shape of placentomes. There was no significant change in the total number of placentomes during gestation. Both mean weight and length increased significantly during gestation, but there was a significant change in the relationship between the two as gestation progressed. There was no correlation between the number of placentomes and the mean weight or length of the placentomes. The development of the placenta in the non-pregnant horn was significantly different from that of the pregnant horn, with fewer, smaller, lighter placentomes. However, there was no evidence that placentome development in the non-pregnant horn was affected by placentome development in the pregnant horn. The proportion of the placentome that was maternal tissue increased during gestation, but there was no significant effect of placentome number, or individual placentome weight or length on this proportion. Two placentome shapes were identified, but no effect on foeto-maternal proportion or relationship between shape and total placentome number or weight was identified.     

Prickly paddy melon (Cucumis myriocarpus) poisoning of cattle

Twenty-six Hereford heifers died after eating mostly ripe fruit of Cucumis myriocarpus growing in a fallowed cultivation paddock. Four affected cattle were dehydrated and apparently had abdominal pain. Necropsy of three revealed intense congestion with haemorrhage of the alimentary tract, numerous C. myriocarpus seeds in ruminal contents, pulmonary congestion and oedema and, in two, swollen livers. Midzonal swelling and vacuolation of hepatocytes occurred in these two. C. myriocarpus fruit (83% by weight ripe) were dosed to two calves at 60 g wet weight/kg live weight. Both collapsed with tachycardia and dyspnoea and died within 6 h. Their packed cell volumes just before death had increased to 0.7. They had hydropic degeneration and necrosis of the ruminal mucosa, intense congestion and oedema of the rumen, abomasum and intestines, swollen and vacuolated hepatocytes and foci of myocardial degeneration and necrosis. Two other calves were dosed daily with 20 g fruit/kg for three days, then 40 g/kg for three days. One calf received a further 40 g/kg next day. Both calves developed persistent diarrhoea and neutrophilia, and their plasma gamma glutamyltransferase and bilirubin concentrations increased. Necropsy revealed necrosis and oedema of the rumen and swollen degenerate hepatocytes.     

α‐6 Integrin Expression in Bovine Spermatogonial Cells Purified by Discontinuous Percoll Density Gradient

The study of spermatogonial stem cells (SSCs) provides a model to better understand adult stem cell biology. Besides the biomedical potential to perform studies of infertility in many species, SSCs hold a promising application at animal transgenesis. Because stem cells are thought to be associated with basement membranes, expression of α‐6 integrin has been investigated as a marker of type A spermatogonial cells, which are considered SSCs because of their undifferentiated status and self‐renewal ability. In this manner, the aim of this study was to isolate type A SSCs from adult bulls by a two‐step enzymatic procedure followed by a discontinuous Percoll density gradient purification and verify the expression of α‐6 integrin by flow cytometry and real‐time RT‐PCR before and after Percoll purification. Spermatogonial cells were successfully obtained using the two‐step enzymatic digestion. An average of 1 × 10⁵ viable cells per gram of testis was isolated. However, the discontinuous Percoll did not purify isolated cells regarding α‐6 integrin expression. Flow cytometry analysis demonstrated no differences in the α‐6 integrin expression between cell samples before and after Percoll purification (p = 0.5636). The same was observed in the real‐time PCR analysis (p > 0.05). In addition to α‐6 integrin, the expression of GFRa‐1 and PGP9.5, known bovine SSCs markers, was detected in all samples studied. Considering that Percoll can reduce cell viability, it is possible to conclude that Percoll density gradient is not suitable to purify bovine SSC, according to α‐6 integrin expression.