Spermatozoa Concentration,Seminal Plasma Composition and Their Physiological Relationship in the Endangered Stellate Sturgeon (Acipenser stellatus) and Russian Sturgeon (Acipenser gueldenstaedtii )

Comparative studies of ionic composition, osmolality, protein concentration and pH of seminal plasma along with spermatozoa concentrations were carried out in stellate sturgeon, Acipenser stellatus, and Russian sturgeon, Acipenser gueldenstaedtii. Analysis of A. gueldenstaedtii sperm showed significantly higher concentrations of Na⁺ (34.58 ± 4.61 mm ), Ca²⁺ (0.35 ± 0.12 mm ), Mg²⁺ (0.70 ± 0.25 mm ), Cl^? (13.50 ± 4.04 mm ) and proteins (0.60 ± 0.29 mg/ml) in the seminal plasma than did seminal plasma of A. stellatus: Na⁺ (20.08 ± 10.75 mm ), Ca²⁺ (0.28 ± 0.06 mm ), Mg²⁺ (0.29 ± 0.05 mm ), Cl^? (7.50 ± 3.00 mm ) and 0.30 ± 0.11 mg/ml proteins. Significantly higher concentration of K⁺ (5.42 ± 1.06 mm ) was observed in A. stellatus compared to A. gueldenstaedtii K⁺ (2.29 ± 0.50 mm ). Concentration of Na⁺ was positively correlated with osmolality (r = 0.819), levels of Cl^? (r = 0.922) and Mg²⁺ (r = 0.727) and pH (r = 0.848). The concentration of Mg²⁺ was positively correlated with protein concentration (r = 0.774), Na⁺ (r = 0.727), Cl^? (r = 0.872) and Ca²⁺ (r = 0.801). A positive relationship was also found between concentration of K⁺ and spermatozoa concentration (r = 0.709). Results revealed strong inter‐species differences in several parameters. The data should be useful for artificial fertilization and for cryopreservation of sturgeon sperm.     

Effects of Hormonal Supplementation on Nuclear Maturation and Cortical Granules Distribution of Canine Oocytes During Various Reproductive Stages

The aim of this study was to evaluate the effects of hCG, progesterone and oestradiol supplementation on nuclear and cytoplasmic maturation of canine oocytes cultured for 24, 48, 72 and 96 h. Oocytes obtained from 18 healthy bitches were divided into three groups according to their reproductive status (follicular, luteal and anoestrus stages) and cultured in TCM 199 + 25 UI/ml of hCG + 1 μg/ml of progesterone + 1 μg/ml of 17‐β oestradiol or without hormonal supplementation (control) for different periods. Then, they were stained with FITC‐LCA‐Hoescht for chromatin configuration and cortical granules distribution and evaluated under an epifluorescence microscope. Culture time and the influence of different stages of the oestrous cycle were also evaluated. The present study demonstrated that there was no significant difference among the reproductive stages. With regards to culture medium, only oocytes from the supplemented medium were able to complete meiosis; however, significant difference was only noticed in the percentage of MI stage oocytes (p < 0.05) in the follicular and luteal group at 72 h of culture. Most oocytes in germinal vesicle, germinal vesicle breakdown and metaphase I stage had cortical granules distributed throughout the cytoplasm (immature pattern), irrespective of the culture period (p < 0.05). Cortical granules distributed immediately beneath the plasma membrane (mature) was only observed in metaphase II stage oocytes, but not all of them presented matured cytoplasm. Our results reveal that cortical granules distribution in canine oocytes matured in vitro did not progressed in correspondence with nuclear stage changes and are in accordance with those from other species.     

GROSS AND HISTOPATHOLOGIC CORRELATION OF LOW‐FIELD MAGNETIC RESONANCE IMAGING FINDINGS IN THE STIFLE OF ASYMPTOMATIC HORSES

With the recent introduction of a 0.25T rotating MRI system, clinical evaluation of the equine stifle joint is now possible in the average equine athlete. A recent publication described common abnormalities of horses with stifle lameness detected with a low‐field MRI system; however, postmortem corroboration of the lesions detected was not possible. Therefore, our objective was to compare postmortem findings with low‐field MRI findings in equine cadaver stifle joints. Ten fresh cadaver stifle joints from horses without clinical signs of stifle disease were evaluated using low‐field MRI, gross dissection, and histopathology. In eight stifles, either the lateral or medial cranial meniscotibial ligament had an irregular shape, fiber separation, or moderate abnormal signal intensity (SI) on all sequences. In five stifles, the medial femoral condyle had articular cartilage fibrillation with or without an osteochondral defect over the weight bearing surface of the medial femoral condyle. All stifles had abnormal SI on all sequences within the patellar ligaments that corresponded with adipose tissue infiltrating between the collagen bundles. Other abnormalities identified included articular cartilage fibrillation of the tibial condyles in three stifles, and articular cartilage fibrillation with chondral defects in the patella in three stifles. All abnormalities detected with low‐field MRI were corroborated by gross dissection. Findings from the current study supported the use of low‐field MRI for detection of stifle joint lesions in horses and demonstrated that some stifle joint pathologies may be subclinical in horses.     

Evaluation of the carryover effect of antibiotic,bee pollen and propolis on growth performance,carcass traits and splenic and hepatic histology of growing rabbits

Sixty‐four nulliparous female rabbits were distributed among eight groups (eight animals/group). Group one was the unsupplemented control group; the other seven groups were supplemented with zinc bacitracin (ZnB) at 100 mg, or bee pollen (BP) and/or propolis (Pro) at 150 and 300 mg in a capsulated form, three times a week, day after day, continuously all over the experimental period. The experiment was run for eight parties; at each parity, 28 kids of each doe group (a total of 224 rabbits) were divided into two subgroups weaned, respectively, at 24 and 30 days of age. Thus, for each parity, there were 16 groups (eight does treatments × two weaning age, 14 rabbits per group). The growing rabbits fed the standard diets without supplements. The growth performance, the carcass traits, the liver and the spleen histology of rabbits were checked up to 90 days of age to find possible carryover effects of the supplements. The supplements had no significant effect on most of the growth performance at 90 days of age, but BP150 and BP+Pro300 increased the growth rate in comparison with ZnB group. The liver weight in the control, BP300 and Pro300 groups was higher than the ZnB one. The spleen weight was higher in the groups ZnB, BP150, Pro300 and BP+Pro300, followed by the control, BP300 and BP+Pro150 and thus Pro150. The heart % in the BP150 and Pro300 groups was higher than ZnB and BP+Pro150 groups. A lymphoid hyperplasia of splenic white pulp was observed in the BP+Pro groups, while propolis alone showed a mild activation of lymphobiosis. The Pro and BP groups showed the same picture of the control group exhibiting a hydropic degeneration of mostly hepatic cells, while the ZnB group exhibited adverse effect on the bile ducts featuring portal periductal inflammatory cells infiltration with epithelial hyperplasia reflecting chronic cholangitis.     

Association of macrophage and lymphocyte infiltration with outcome in canine osteosarcoma

Immunotherapeutic strategies have shown promise for the treatment of canine osteosarcoma (cOSA). Very little is known about the immune microenvironment within cOSA, however, limiting our ability to identify potential immune targets and biomarkers of therapeutic response. We therefore prospectively assessed the disease‐free interval (DFI) and overall survival time (ST) of 30 dogs with cOSA treated with amputation and six doses of adjuvant carboplatin. We then quantified lymphocytic (CD3+, FOXP3+) and macrophage (CD204+) infiltrates within the primary tumours of this cohort using immunohistochemistry, and evaluated their association with outcome. Overall, the median DFI and ST were 392 and 455 days, respectively. The median number of CD3+ and FOXP3+ infiltrates were 45.8 cells/mm² (4.6‐607.6 cells/mm²) and 8.5 mm² (0‐163.1 cells/mm²), respectively. The median area of CD204+ macrophages was 4.7% (1.3%‐23.3%), and dogs with tumours containing greater than 4.7% CD204+ macrophages experienced a significantly longer DFI (P = 0.016). Interestingly, a significantly lower percentage of CD204+ macrophages was detected in cOSA arising from the proximal humerus compared to other appendicular bone locations (P = 0.016). Lymphocytic infiltrates did not appear to correlate with outcome in cOSA. Overall, our findings suggest that macrophages may play a role in inhibiting cOSA progression, as has been suggested in human osteosarcoma.     

Ultra‐frequent HRAS p.Q61R somatic mutation in canine acanthomatous ameloblastoma reveals pathogenic similarities with human ameloblastoma

Ameloblastoma is a locally aggressive odontogenic tumour that occurs in humans and dogs. Most ameloblastomas (AM) in humans harbour mutually‐exclusive driving mutations in BRAF, HRAS, KRAS, NRAS or FGFR2 that activate MAPK signalling, and in SMO that activates Hedgehog signalling. The remarkable clinical and histological similarities between canine acanthomatous ameloblastoma (CAA) and AM suggest they may harbour similar driving mutations. In this study, aimed at characterizing the mutational status of SMO, BRAF, HRAS, KRAS, NRAS and FGFR2 in CAA, we used RNA sequencing, Sanger sequencing and restriction fragment length polymorphism assays to demonstrate that 94% of CAA (n = 16) harbour a somatic HRAS p.Q61R mutation. The similarities in MAPK‐activating mutational profiles between CAA and AM implicate conserved molecular mechanisms of tumorigenesis, thus, qualifying the dog as a potentially useful model of disease. Given the relevance of RAS mutations in the pathogenesis of odontogenic tumours and other types of cancer, the results of this study are of comparative, translational, and veterinary value.