Evaluation of numbers of microscopic and macroscopic follicles in cattle selected for twinning

We hypothesized that the number of microscopic follicles present in the ovaries of cattle selected for twin births (Twinner) would be greater than in the ovaries of contemporary Controls. Ovaries were collected from seven Control and seven Twinner cows at slaughter. The number of Small (1 to 3.9 mm), Medium (4 to 7.9), and Large (> 8 mm) surface follicles was counted and one ovary was fixed for histological evaluation. Fifty to sixty consecutive 6-microm slices were taken from a piece of cortical tissue, approximately 1 cm x 1 cm in area, located between the surface follicles. Microscopic follicles were classified as primordial (oocyte surrounded by a single layer of squamous pregranulosa cells), primary (oocyte surrounded by a single layer of one or more cuboidal granulosa cells), secondary (oocyte surrounded by two or more layers of granulosa cells), or tertiary (oocyte surrounded by multiple layers of granulosa cells with initiation of antrum formation to < or = 1 mm in diameter). The total number of follicles was counted in 200 fields (2 mm x 2 mm) per ovary. A field containing no follicles was classified as empty. There were significantly more secondary follicles in Twinner compared with Control ovaries (12.9 vs 6.3; P < .05). Twinners also tended to have more small surface follicles (35.4 vs 49.0; P < 0.1). We conclude that ovaries of Control and Twinner cows do not differ in the number of primordial follicles or in the number of follicles activated into the growing pool; however, Twinner cows are able to maintain more growing follicles at the secondary and subsequent stages of development.     

Grafts of porcine small intestinal submucosa seeded with cultured homologous smooth muscle cells for bladder repair in dogs

Background

Due to numerous complications associated to gastrointestinal augmented cystoplasty, this study aimed to analyze the anatomic repair of the bladder of 10 female dogs using grafts of porcine small intestinal submucosa (SIS) seeded with cultured homologous smooth muscle cells, and compare them with the acellular SIS grafts.

Results

We assessed the possible side effects and complications of each type of graft by clinical examination, abdominal ultrasound and laboratory findings. Anatomic repair of neoformed bladder was assessed by histological staining for H/E and Masson''s Trichrome, analyzed with a Nikon Photomicroscope connected to the system of image analysis Image J.

Conclusions

We propose that SIS associated to homologous smooth cells can improve the quality of tissue repair, and consequently decrease the potential complications inherent to acellular SIS.     

Expression and localization of angiopoietin family in corpus luteum during different stages of oestrous cycle and modulatory role of angiopoietins on steroidogenesis,angiogenesis and survivability of cultured buffalo luteal cells

The objective of this study was to document the expression and localization of angiopoietin (ANGPT) family members comprising of angiopoietin (ANGPT1 and ANGPT2), and their receptors (Tie1 and Tie2) in buffalo corpus luteum (CL) obtained from different stages of the oestrous cycle, and the modulatory role of ANGPT1 and ANGPT2 alone or in combinations on progesterone (P₄) secretion and mRNA expression of phosphotidylinositide‐3kinase‐protein kinase B (PI3K‐AKT), phosphoinositide‐dependent kinase (PDK), protein kinase B (AKT), Bcl2 associated death promoter (BAD), caspase 3 and von willebrand factor (vWF) in luteal cells obtained from midluteal phase (MLP) of oestrous cycle in buffalo. Real‐time RT‐PCR (qPCR), Western blot and immunohistochemistry were applied to investigate mRNA expression, protein expression and localization of examined factors whereas, the P₄ secretion was assessed by RIA. The mRNA and protein expression of ANGPT1 and Tie2 was maximum (p < .05) in mid luteal phase (MLP) of oestrous cycle. The ANGPT2 mRNA and protein expression was maximum (p < .05) in early luteal phase, decreased in MLP and again increased in late luteal phase of oestrous cycle. ANGPT family members were localized in luteal cells and endothelial cells with a stage specific immunoreactivity. P₄ secretion was highest (p < .05) with 100 ng/ml at 72 hr when luteal cells were treated with either protein alone. The mRNA expression of PDK, AKT and vWF was highest (p < .05) and BAD along with caspase 3 were lowest (p < .05) at 100 ng/ml at 72 hr of incubation period, when cultured luteal cells were treated with either protein alone or in combination. To conclude, our study explores the steroidogenic potential of angiopoietins to promote P₄ secretion, luteal cell survival and angiogenesis through an autocrine and paracrine actions in buffalo CL.     

Influence of vitamin A injection before mating on oocyte development, follicular hormones, and ovulation in gilts fed high-energy diets

Previous research revealed that treatment with vitamin A approximately 5 d before ovulation may increase litter size in weaned sows and improve embryonal survival in gilts fed high-energy diets that reduced embryonal survival. For the current study, the hypothesis was that administration of vitamin A before ovulation would alter development of follicles and oocytes in a way favorable to enhanced embryonal survival. (Landrace x Large White) x (Duroc x Hampshire) gilts (n = 44) were fed 11.0 Mcal ME x gilt(-1) x d(-1) beginning 7 d after second estrus and given (i.m.) corn oil or 1 x 10(6) IU of vitamin A (retinyl palmitate) on d 15 after second estrus. Gilts were checked for estrus every 4 h, mated naturally at third estrus, and assigned randomly to undergo midventral laparotomy beginning at 24 to 28, 28 to 32, 32 to 36, or 36 to 40 h after onset of third estrus. At laparotomy, ovulated oocytes and early-stage embryos were recovered from oviducts, and ovaries were removed for aspiration of oocytes and granulosa cells from unovulated follicles. Oocytes and embryos were stained for assessment of stage of development. Granulosa cells were cultured to assess their ability to secrete progesterone. Follicular fluid was assayed for progesterone, estradiol-17beta, IGF-I, and PGF2alpha. Treatment with vitamin A altered development of oocytes and embryos by decreasing the percentage at the germinal vesicle stage and increasing the percentage at advanced stages. Mean stage of development was increased by vitamin A, but variation in stage was decreased. Among follicles matched by meiotic stage of oocyte, follicular fluid concentrations of progesterone, IGF-I, and PGF2alpha were greater in vitamin A-treated gilts than in controls, but treatment with vitamin A in vivo did not affect LH-stimulated or unstimulated secretion of progesterone by granulosa cells in vitro. These data provide evidence that vitamin A may influence embryonic development by advancing resumption of meiosis and altering follicular hormonal environment during follicle maturation.     

DIAGNOSIS OF AORTIC THROMBOEMBOLISM IN TWO DOGS WITH RADIONUCLIDE ANGIOGRAPHY

Radionuclide angiography was used to document occlusion of the distal aorta by thromboemboli in two dogs. Findings were confirmed by necropsy. The location of the thrombus corresponded with the scintigraphic lesion. Information obtained by the radionuclide angiogram was instrumental in patient management. We suggest the use of radionuclide angiography as a quick, non-invasive alternative to contrast angiography in animals suspected of having aortic thromboembolism.