Organic zinc and copper supplementation on antioxidant protective mechanism and their correlation with sperm functional characteristics in goats

Trace minerals feeding had significant effects on sperm production and fertility with better absorption and proper utilization within the body for optimum reproductive function. Several studies have shown that more influenced trace elements in the diets of animals are copper (Cu) and zinc (Zn). Bucks showing deficiency of this mineral might affect the quality of semen production which in turn would affect the fertility. This experiment was thus designed to test the effects of organic Cu and Zn supplementation on antioxidants enzyme activities and sperm functional attributes in fresh semen of bucks. Forty bucks (n = 40, Aged 5 months) were assigned to ten groups of four animals in each group, supplemented (for a period of 8 months) with different levels of organic Zn: 20 mg (T2), 40 mg (T3) and 60 mg (T4), organic Cu: 12.5 mg (T5), 25 mg (T6), 37.5 mg (T7) and combined organic Zn and Cu: 20 + 12.5 mg (T8), 40 + 25 mg (T9), 60 + 37.5 mg (T10), respectively, per kg dry matter and no additional mineral diet (control; T1). One hundred and sixty semen samples were collected through electro‐ejaculator and analysed for sperm quantity, quality, acrosome intactness and plasma membrane integrity and correlated with the catalase, superoxide dismutase, glutathione peroxidase and glutathione reductase enzyme activities in seminal plasma. The results indicated organic Cu and zinc supplemented bucks produced more sperm cells, had higher sperm concentrations, maintained higher (p < .01) sperm livability, plasma membrane and acrosome integrities, more motility and velocity. The increased antioxidant enzyme activities, reduced oxidative stress and lowered lipid peroxidation were positively correlated (p < .05) with the sperm functional attributes. In conclusion, organic Cu and Zn supplement to male goats showed protective roles against oxidative damage and maintained better fresh semen characteristics.     

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.     

Production of Wild Buffalo (Bubalus arnee) Embryos by Interspecies Somatic Cell Nuclear Transfer Using Domestic Buffalo (Bubalus bubalis) Oocytes

The objective of this study was to explore the possibility of producing wild buffalo embryos by interspecies somatic cell nuclear transfer (iSCNT) through handmade cloning using wild buffalo somatic cells and domestic buffalo (Bubalus bubalis) oocytes. Somatic cells derived from the ear skin of wild buffalo were found to express vimentin but not keratin and cytokeratin‐18, indicating that they were of fibroblast origin. The population doubling time of skin fibroblasts from wild buffalo was significantly (p < 0.05) higher, and the cell proliferation rate was significantly (p < 0.05) lower compared with that of skin fibroblasts from domestic buffalo. Neither the cleavage (92.6 ± 2.0% vs 92.8 ± 2.0%) nor the blastocyst rate (42.4 ± 2.4% vs 38.7 ± 2.8%) was significantly different between the intraspecies cloned embryos produced using skin fibroblasts from domestic buffalo and interspecies cloned embryos produced using skin fibroblasts from wild buffalo. However, the total cell number (TCN) was significantly (p < 0.05) lower (192.0 ± 25.6 vs 345.7 ± 42.2), and the apoptotic index was significantly (p < 0.05) higher (15.1 ± 3.1 vs 8.0 ± 1.4) for interspecies than that for intraspecies cloned embryos. Following vitrification in open‐pulled straws (OPS) and warming, although the cryosurvival rate of both types of cloned embryos, as indicated by their re‐expansion rate, was not significantly different (34.8 ± 1.5% vs 47.8 ± 7.8), the apoptotic index was significantly (p < 0.05) higher for vitrified–warmed interspecies than that for corresponding intraspecies cloned embryos (48.9 ± 7.2 vs 23.9 ± 2.8). The global level of H3K18ac was significantly (p < 0.05) lower in interspecies cloned embryos than that in intraspecies cloned embryos. The expression level of HDAC1, DNMT3a and CASPASE3 was significantly (p < 0.05) higher, that of P53 was significantly (p < 0.05) lower in interspecies than in intraspecies embryos, whereas that of DNMT1 was similar between the two types of embryos. In conclusion, these results demonstrate that wild buffalo embryos can be produced by iSCNT.     

‘Soya Milk Tris‐based Phytoextender Reduces Apoptosis in Cryopreserved Buffalo (Bubalus bubalis) Spermatozoa’

The objective of this study was to investigate the effect of newly developed soya milk Tris (SMT)‐based phytoextender as an alternative to egg yolk Tris (EYT) extender used for cryopreservation of buffalo (Bubalus bubalis) spermatozoa on apoptosis. Fresh buffalo semen (control without dilution) was cryopreserved in conventional EYT (20% egg yolk v/v in Tris) and SMT (25% soya milk v/v in Tris) extender and used for the assessment of expression of apoptotic proteins. Proteins extracted from a total number of nine ejaculates from three individual buffalo bulls chosen at random were separated using SDS–PAGE followed by immunoblotting against caspase‐8, caspase‐9, caspase‐3, poly(ADP‐ribose)polymerase (PARP), cytochrome c and apoptosis inducing factor (AIF). In addition, fluorescence microscopy was used for the detection of mitochondrial membrane potential (JC‐1 assay) and apoptotic cells (annexin V‐FITC/PI assay). The results obtained clearly indicate the significant (p < 0.05) reduction in the expression of caspase‐3 (27 kDa), caspase‐8 (53 kDa), caspase‐9 (50 kDa) precursor and cytochrome c (17 kDa) in semen cryopreserved in SMT extender in comparison with EYT extender. A non‐significant (p > 0.05) reduction in expression of PARP‐DNA‐binding subunit (24 kDa) was observed in SMT extender. No expression of AIF was found in cryopreserved semen samples. A significant (p < 0.05) increase in the mean percentage of cells having high mitochondrial membrane potential and a non‐significant (p > 0.05) decrease in late apoptotic cells (AN+/PI+) was observed in SMT extender when compared to EYT extender. The results demonstrated that cryopreservation of buffalo semen in SMT‐based phytoextender can replace the traditional egg yolk extenders as it reduces the expression of apoptotic proteins maintaining high mitochondrial membrane potential and gives better protection to sperms in terms of its non‐animal origin.     

Treatment of rupture of suspensory ligament and superficial flexor tendon in a bull

Rupture of the suspensory ligament at the insertions on the proximal sesamoid bones, and of the superficial flexor tendon of the left fore limb, occurred in an adult Angus bull as a result of fighting. There was severe hyperextension of the metacarpophalangeal (MCP) joint with the dewclaws almost touching the ground. Radiographs revealed severe hyper-extension of the MCP joint with the sesamoid bones aligned directly distal to the metacarpus. Initially, a full length fiberglass cast was applied with the limb partially flexed within the cast and the heels elevated. The cast was replaced twice. The cast was removed after 136 days and the bull was bearing full weight on the limb. Prolonged immobilisation of the limb produced new bone in the area (a normal response in cattle) to cause ankylosis of the traumatized MCP joint and partial ankylosis of the carpus. The bull was being used for pasture breeding one year after the injury.     

不同年代大豆品种(系)根系生长动态的比较研究

不同年代大豆品种根干重积累动态可以用Logistic曲线方程进行模拟。在不同施肥水平下,根干重最大增长速率及出现的时间和平均增长速率有所不同。根干重最大增长速率出现的时间,中国当代品种、美国当代品种及中国20世纪20年代的品种间差异不显著,各品种间差异及施肥处理间差异也不显著。根干重最大增长速率,中国当代、美国当代及中国20世纪20年代的品种间差异不显著,各品种间差异也不显著,施肥处理间差异显著。平均增长速率,中国当代、美国当代及中国20年代的品种间差异显著,各品种间及施肥处理间差异也显著。不同施肥水平、不同生育时期,中国当代品种与美国当代品种根干重都比中国20年代品种大。     

The experimental production of big liver and spleen disease in broiler breeder hens

SUMMARY Big liver and spleen disease (BLS) was reproduced experimentally by intravenous (IV) and oral (PO) administration of BLS inocula to susceptible broiler breeder hens 34 to 36 weeks of age. Serological and pathological signs of BLS similar to those seen in the natural disease occurred in inoculated and in-contact birds. Splenomegaly was the earliest and often the only necropsy finding, with hepatomegaly and kidney enlargement occurring in some birds later in the course of the disease. After IV administration, serum antigen was detected between 2 and 4 weeks, and antibody between 3 and 5 weeks. After PO administration, antigen was detected between 2 and 4 weeks, and antibody between 3 and 6 weeks. Antibody persisted in all birds to the end of the experiment (6 weeks), and horizontal transmission probably occurred since in-contact birds developed BLS. Liver probably contained the highest concentration of BLS agent because it had the highest infectivity.     

The Effects of the Periodical Use of In-feed Chlortetracycline on the Reproductive Performance of Gilts and Sows of a Commercial Pig Farm with a History of Clinical and Subclinical Viral and Bacterial Infections

The objective of this study was to assess the effects of in‐feed chlortetracycline (CTC) as a measure of preventing or minimizing infectious problems of reproductive failure in gilts and sows. In a farm of 400 Large White × Landrace gilts and sows with a clinical history of porcine respiratory and reproductive syndrome (PRRS) virus, the animals were treated with CTC. Treatment consisted of 10 g CTC sow/day for 15 days every 3 months. It improved the health status of sows by decreasing post‐farrowing clinical mastitis and vaginal discharges, abortions, return‐to‐oestrus and irregular return‐to‐oestrus rates. These beneficial effects had a positive impact on the performance of the litter. More piglets were born live and weaned. These positive effects improved with repeated use of CTC. The serological evidence of PRRS virus, Leptospira spp. and Chlamydia spp. and the subsequent beneficial use of the antimicrobial agent indicate that reproductive failure, possibly resulting from the bacterial agents can be controlled with in‐feed use of broad spectrum antimicrobials.     

Structural and physiological changes associated with the skin spot disorder in apple

Skin spot is an important physiological disorder of ‘Elstar’ apples (Malus × domestica Borkh.) that occurs after fruit have been removed from controlled atmosphere storage. Skin spots are irregular patches of small, round, brown blemishes. Cross-sections reveal a browning of protoplasts (coagulated) and of cell walls that extends into the hypodermis. Skin spots are always associated with linear, gaping and non-gaping microcracks in the cuticle. Staining of apple skin with calcofluor white usually results in white fluorescence of cell walls but, within a skin spot, the white fluorescence is weak or absent. Cell walls within, and in the immediate vicinity of skin spots stain with phloroglucin/HCl indicating the presence of lignin. The area of skin affected by skin spots was positively and linearly correlated with the area of the non-blush fruit surface infiltrated by acridine orange. In general, skin spots were limited to the non-blush fruit surface and occurred more frequently near the stem-end than the calyx region of the fruit. Skin spot areas were correlated with a 2.5-fold increase in water vapour permeability compared with unaffected areas (23.8 ± 4.0 m s⁻¹ with skin spots, 9.6 ± 2.1 × 10⁻⁵ m s⁻¹ without skin spots). Strips of the fruit skin from regions with skin spots had an increased maximum force and modulus of elasticity. Dipping fruit in ascorbic acid (0.1 or 0.3 mM for 10 min) before storage decreased the area affected by skin spots. There was no effect of dipping in ethanol/water (70%, v/v, 15 min) or in solutions of captan (1.5 g L⁻¹, 10 min) or trifloxystrobin (0.1 g L⁻¹, 10 min). In contrast, prestorage treatment with 1-methylcyclopropene (630 nL L⁻¹ for 24 h) or poststorage incubation in H₂O₂ (10% for 2, 6, 10 and 16 h) increased skin spots. Our data are consistent with a typical cell response to an oxidative burst that seems to be focussed on particular regions of the ‘Elstar’ fruit surface by concentrations of cuticular microcracks, and that is possibly caused by reoxygenation injury upon removal from CA storage.