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.     

Effect of dietary protein on intake,nutrients utilization,nitrogen balance,blood metabolites,growth and puberty in growing Bhadawari buffalo (Bubalus bubalis) heifers

Fifteen Bhadawari buffalo heifers of 207?±?9.78 kg mean body weight were randomly distributed into three dietary groups to evaluate the effect of protein level on nutrient utilization, nitrogen (N) balance, growth rate, blood metabolites, and puberty. All animals were offered wheat straw-berseem diets supplemented with concentrate mixtures of similar energy (2.7 Mcal/kg) and different protein levels (14.3–22 %). Animals of standard-protein group (SPG) were offered protein and energy as per requirement, while animals of low-protein group (LPG) and high-protein group (HPG) were fed 20 % less and 20 % more protein, respectively, than SPG. Feed dry matter (DM) and metabolizable energy (ME) intake (% body wt. and g/kg w^0.75) were similar for all three diets; however, the crude protein (CP) and digestible crude protein (DCP) intake on percent body weight and per kilogram metabolic weight was higher (P?<?0.05) in HPG than in SPG or LPG. Digestibility of CP, cellulose, and hemicellulose was higher (P?<?0.05) in HPG versus LPG. Fecal N excretion was similar, while urinary N excretion was highest (P?<?0.05) in HPG (74.83 g/day) compared with SPG (50.03 g/day) and LPG (47.88 g/day), which resulted in lower N retention in HPG than in the other dietary groups. Level of dietary N had no effect on blood metabolites viz. glucose, urea, and N. Digestible energy (DE) and ME contents of diets were identical, while DCP contents were higher (P?<?0.05) in HPG than in LPG. Feed and nutrient (CP and ME) conversion efficiency to produce a unit kilogram weight gain was identical among the dietary groups. Dietary protein level had no effect on the heifer’s weight and age at puberty. The mean growth rate of heifers at 240 days was higher (P?>?0.05) in SPG (330.8 g/day) than in LPG (296.7 g/day), while the animals gained more weight in January to March months and the lowest weight in May to July months. Protein level had no effect on conception rate of heifers. Results revealed that 20 % higher or less protein than the ICAR requirement had no significant (P?>?0.05) on feed intake, nutrient conversion efficiency for weight gain, heifer growth, and puberty; however, 20 % more protein increased urinary N loss.

     

Development of a murine ocular posterior segment explant culture for the study of intravitreous vector delivery

The objective of this study was to develop a murine retinal/choroidal/scleral explant culture system to facilitate the intravitreous delivery of vectors. Posterior segment explants from adult mice of 2 different age groups (4 wk and 15 wk) were cultured in serum-free medium for variable time periods. Tissue viability was assessed by gross morphology, cell survival quantification, activated caspase-3 expression, and immunohistochemistry. To model ocular gene therapy, explants were exposed to varying transducing units of a lentiviral vector expressing the gene for green fluorescent protein for 48 h. Explant retinal cells remained viable for approximately 1 wk, although the ganglion cell layer developed apoptosis between 4 and 7 d. Following vector infusion into the posterior segment cups, viral transduction was noted in multiple retinal layers in both age groups. An age of donor mouse influence was noted and older mice did not transduce as well as younger mice. This explant offers an easily managed posterior segment ocular culture with minimum disturbance of the tissue, and may be useful for investigating methods of enhancing retinal gene therapy under controlled conditions.     

Comparative Expression Analysis of Gametogenesis‐Associated Genes in Foetal and Adult Bubaline (Bubalus bubalis) Ovaries and Testes

This study was conducted to identify and analyse the expression of gametogenesis‐associated genes and proteins in foetal and adult buffalo gonads of both the sexes. Relative quantification of the genes was determined by qPCR and Western blotting. Immunohistochemistry was also performed for various gametogenesis‐associated proteins in foetal and adult gonads of both the sexes. We observed significantly (p < 0.05) increased expression of primordial germ cell‐specific, meiotic as well as genes associated with oocyte maturation and development in foetal ovaries as compared to the adult ones. However, significantly (p < 0.05) increased expression of proteins associated with oocyte maturation like GDF9 and ZP4 was found in adult ovaries, indicating temporal regulation of mRNA translation during oogenesis. Meiotic genes showed significantly (p < 0.05) increased expression in adult testes as compared to foetal testes and ovaries, indicating onset of meiosis at a later stage in spermatogenesis. In general, the expression of primordial germ cell‐associated as well as meiotic genes was higher in adult testes, indicating the increased biological activity in the organ. Immunohistochemistry revealed localized expression of gametogenesis‐associated proteins in ovarian follicles and seminiferous tubules of testes, while the surrounding somatic tissues were devoid of these proteins. The study gives an understanding of the sequential and temporal events of gene expression as well as mRNA translation during male and female gametogenesis. It could also be concluded that follicles and seminiferous tubules are the functional units of the female and male gonads, respectively, and their function could be enhanced by appropriate chemical and genetic intervention of the somatic tissue immediately surrounding them. This assumes importance in the context that buffalo attains sexual maturity at an older age of 2–3 years and have smaller ovaries with lesser number of primordial follicles in comparison with cattle, which is suggested to be the main reason of their poor breeding performance.     

Toxicological effects and recovery of the corneal epithelium in Cyprinus carpio communis Linn. exposed to monocrotophos: an scanning electron microscope study

This study was conducted based on the evidence of fish habitats in North India being affected by organophosphate pesticides draining from agricultural fields into bodies of water, especially during the rainy season. Various tissues of fish such as scales, gills ovaries, kidney, and liver have been studied from the toxicological point of view, but the toxicological effects of aquatic pollutants on fish cornea have not been investigated to date. We conducted comparative toxicological studies on the cornea of Cyprinus carpio communis using two sublethal (0.038 and 0.126 ppm) concentrations of monocrotophos pesticide for 30 days. Corneas from all the groups were evaluated by a scanning electron microscope. The fish exposed to the monocrotophos pesticide developed corneal necrosis due to the formation of crystalloid‐like structures, thinning and shrinkage of microridges on the corneal epithelium. After 30 days, fish from the monocrotophos‐treated tank were transferred to normal environmental conditions. After 60 days under natural condition, epithelial cells did not fully recover. In conclusion, exposure to monocrotophos induces irreversible changes in the cornea of C. carpio communis. As fish and mammalian visual systems share many similarities, the reported finding may offer useful insights for further toxicological and ophthalmological studies in humans.     

轴向间距对贯流式水轮机水力性能的影响

为了寻找贯流式水轮机中导叶与转轮轴向间距的合理取值,在保证其结构紧凑性的同时,获得良好的水力性能,在其他主要参数不变的情况下,通过试验与数值计算相结合的方法,分别对导叶与转轮之间轴向间距s值为30, 40, 50, 60和70 mm 的微贯流式水轮机模型进行全流场数值计算,研究轴向间距对贯流式水轮机水力性能的影响.结果表明:水轮机的水头随着轴向间距的增大而增加,水轮机的轴功率随着轴向间距的增大呈现先上升后稳定的趋势;综合各流量条件,水轮机通常在适中的轴向间距下取得较高的效率,该模型在s=40 mm时效率达到最高.选择适中的轴向间距可以改善转轮叶片表面压力分布和流态;过大的轴向间距会导致转轮进口发生负撞击,出现脱流现象;转轮出口的湍动能随着轴向间距的扩大而减小.在最优工况下,轴向间距从30 mm扩大至70 mm的过程中,转轮内部的水力损失逐渐增加,增加幅度达到15.6%,而出水管中的水力损失则减小了19.7%,总水力损失呈增加趋势.综合考虑,选用s=40 mm 作为最终方案.     

Binding of 2-nonanone and milk proteins in aqueous model systems

Interactions of the model flavor compound 2-nonanone with individual milk proteins, whey protein isolate (WPI), and sodium caseinate in aqueous solutions were investigated. A method to quantify the free 2-nonanone was developed using headspace solid-phase microextraction followed by gas chromatography with flame ionization detection. Binding constants (K) and numbers of binding sites (n) for 2-nonanone on the individual proteins were calculated. The 2-nonanone binding capacities decreased in the order bovine serum albumin > beta-lactoglobulin > alpha-lactalbumin > alpha s1-casein > beta-casein, and the binding to WPI was stronger than the binding to sodium caseinate. All proteins appeared to have one binding site for 2-nonanone per molecule of protein at the flavor concentrations investigated, except for bovine serum albumin, which possessed two classes of binding sites. The binding mechanism is believed to involve predominantly hydrophobic interactions.     

Changes in tissue lipid levels in the freshwater catfishClarias batrachus associated with the reproductive cycle

The levels of free fatty acid (FFA), monoglyceride (MG), diglyceride (DG), triglyceride (TG), phospholipid (PL), free cholesterol (CF) and esterified cholesterol (CE) were measured in the liver, plasma, ovary, abdominal fat and muscle during different phases of the annual reproductive cycle in femaleClarias batrachus. During the preparatory phase, hepatic lipogenic activity predominated over mobilization and consumption. In the prespawning season, an increased hepatic lipogenic activity was maintained, but lipids were transferred from the liver to the ovary. In the spawning phase, the diminished food intake, and enhanced caloric demand for spawning behaviour and activity limited hepatic lipogenic activity, and TG lypolysis was increased as was the production of more FFA. Maximum accumulation of vitellogenin, as reflected by maximum rise in ovarian PL titre was characteristic of this phase. Marked reductions in ovarian lipid occurred during the postspawning phase. In the resting phase, there was a recovery of lipogenic activity, but PL synthesis was still inhibited. In contrast to other investigated teleosts, there were extremely high level of FFA in liver, plasma, ovary and muscle throughout the annual reproductive cycle inC. batrachus. FFA appears to be the main lipid metabolite which had a very high turnover. As evidenced by the high TG content, abdominal fat seems to be the main fat depot, not the liver and muscle.Correspondent author     

Role of cathepsins B and D in proteolysis of yolk in the catfish Clarias gariepinus

Fish Physiology and Biochemistry - Yolk processing pathways vary in the oocytes of benthophil and pelagophil teleosts. The present study investigated the yolk processing pattern in the oocytes of...     

Effect of cryoprotectants, equilibration periods and freezing rates on cryopreservation of spermatozoa of mahseer, Tor khudree (Sykes) and T. putitora (Hamilton)

A study was conducted to standardize a protocol for cryopreservation of spermatozoa of the endangered mahseer, Tor khudree (Sykes) and T. putitora (Hamilton). The suitability of the cryoprotectants, dimethyl sulphoxide (DMSO) and glycerol, and the combination of the two were tested. Four equilibration periods and four freezing rates were also tested for their standardization. A combination of 9% DMSO and 11% glycerol gave significantly higher mean percentage of hatching in both T. khudree (45.59±1.86%) (control 71.08±0.59%) and T. putitora (45.00±1.25%) (control 73.48±1.19%) among the eight different treatments. Among the four different equilibration periods tested, the equilibration period of 30 min^?1 yielded the highest mean hatching percentage in T. khudree (39.46±1.94%) (control 71.70±0.61%) and T. putitora (38.28±1.06%) (control 73.11±0.82%). Freezing straws at a height of 8 cm above LN₂ surface for 10 min^?1 gave higher hatching percentages for both T. khudree (41.75±1.72%) (control 73.99±1.17%) and T. putitora (41.34±2.04%) (control 72.48±1.51%). The study reports the superior performance of the combination of DMSO and glycerol for the first time.