Ribotyping of Indian Isolates of <Emphasis Type="Italic">Pasteurella multocida</Emphasis> Based on 16S and 23S rRNA Genes

The applicability of ribotyping based on 16S and 23S rRNA was evaluated for molecular epidemiological studies. Forty-eight isolates of Pasteurella multocida isolated from different hosts and geographical locations and one reference isolate were ribotyped. Only four ribotypes were found. All the isolates including reference isolate from wild carnivores had the same ribotype, though they had different serotypes. The isolate from a tiger had one band in addition to the bands present in the major ribotype. The isolates from lions represented two ribotypes; of these ribotypes, one (r2) had an additional band of 3.6 kbp, which was absent in all other ribotypes. The second ribotype (r4) from a lion had one band missing (6 kbp) that was present in the other ribotypes. These isolates were further typed using ERIC-PCR and REP-PCR. With ERIC-PCR and REP-PCR, higher D values of 0.83 and 0.89 were obtained. The current study revealed that ribotyping is not a very efficient typing tool for use in molecular epidemiology for differentiation of isolates.     

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.     

CONTRAST RADIOGRAPHY OF THE OVINE GASTROINTESTINAL TRACT

Contrast radiography of the alimentary canal was conducted in six adult sheep with barium sulfate (70% W/V, 25–30 ml/kg). A period of 12 hours was adequate for opacification of most parts of the gastrointestinal tract. The presence of contrast medium in different parts of the alimentary canal was noted for as long as 60 hours. The right lateral view was adequate for visualization of most structures, but the typical laminar mucosal pattern of the omasum and the spiral mucosal folds of the abomasum were demonstrated better on the ventrodorsal view.     

Effect of Supplementation of Taurine or Trehalose in Extender on Immunolocalization of Tyrosine Phosphoproteins in Buffalo and Cattle (Karan Fries) Cryopreserved Spermatozoa

The present study assessed the effects of incorporation of Taurine or Trehalose in extender on immunolocalization of tyrosine phosphoproteins, Cryocapacitation and other sperm quality parameters (motility, viability and membrane integrity) in post‐thawed sperm from Buffalo (Murrah) and Cattle (Karan Fries). Six ejaculates from six individual bulls from both species were chosen at random and split into four aliquots: one aliquot without dilution (fresh sample), another diluted in egg yolk tris‐citrate (EYTC) extender and the rest of aliquots with EYTC dilution supplemented with taurine (50 mm ) or trehalose (100 mm ), respectively, and cryopreserved. Following cryopreservation, semen were thawed and assessed for standard semen quality parameters. Extent of capacitation in cryopreserved spermatozoa was measured by inducing in vitro acrosome reaction followed by dual staining. Immunolocalization of tyrosine phosphoproteins was carried out by immunocytochemistry using primary antibody clone pT‐154 (anti‐phosphotyrosine antibody) and FITC‐conjugated secondary antibody. Immunofluorescent signals were analysed for level of protein tyrosine phosphorylation in spermatozoa. Post‐thaw semen evaluation showed supplementation of taurine or trehalose to EYTC extender significantly (p < 0.05) increased motility, viability and membrane integrity of spermatozoa in both species. Percentage of cryocapacitated spermatozoa was significantly (p < 0.05) higher in cattle as compared to buffalo and degree of cryocapacitaion of spermatozoa decreased significantly (p < 0.05) upon supplementation of additives in both the species. It was also found that tyrosine phosphoproteins were localized differentially in fresh and cryopreserved spermatozoa. Supplementation of taurine or trehalose to freezing extender changed the localization of tyrosine phosphoproteins in cryopreserved spermatozoa similar to fresh in both the species. The results obtained clearly indicated that supplementation of taurine or trehalose to EYTC prior to cryopreservation improves Buffalo and Cattle sperm quality in terms of cryocapacitation and immunolocalization of tyrosine phosphoproteins during freezing–thawing process.     

Status of selenium and vitamin E on Norwegian organic sheep and dairy cattle farms

Herbage selenium (Se) concentration is generally low in Norway. It is unknown whether feeding practices on Norwegian organic farms fulfil the dietary needs of Se and vitamin E for sheep and dairy cattle. Therefore we analysed Se in soil and herbage, and Se and vitamin E in animal blood in the indoor feeding season at 14 organic dairy and 14 organic sheep farms. The herbage Se concentration was low. Approximately 50 and 35% of all samples in the first and second cut, respectively, had Se concentrations below the detection limit of 0.01 mg/kg dry matter (DM). The median (10th, 90th percentile) Se concentrations were <0.01 (<0.01, 0.03) and 0.02 (<0.01, 0.06) mg/kg DM in the first and second cuts, respectively. Whole blood Se concentrations were 0.10 (0.04, 0.15) μg/g in dairy cattle and 0.14 (0.03, 0.26) μg/g in sheep. Vitamin E concentrations were 4.2 (2.7, 8.4) mg/l in dairy cattle and 1.3 (0.9, 2.4) mg/l in sheep. None of the soil or plant variables explained the variation in herbage Se concentration, although Se in soil and plant tended to be correlated. Herbage Se concentration was inadequate to meet the dietary Se requirements. Vitamin E requirement was only met in dairy herds. We recommend Se and vitamin E supplementation to ruminants on organic farms.     

Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers

True-to-type clonal fidelity is one of the most important pre-requisites in micropropagation of crop species. Genetic fidelity of in vitro raised 45 plants of gerbera (Gerbera jamesonii Bolus) derived from three different explants, viz., capitulum, leaf and shoot tips, was assessed by 32 ISSR markers, for their genetic stability. Out of 32 ISSR markers, 15 markers produced clear, distinct and scorable bands with an average of 5.47 bands per marker. The markers designed from AG motif amplified more number of bands. The markers anchored at 3′ ends produced high number of consistent bands than unanchored markers. Fifteen ISSR markers generated a total of 3773 bands, out of which 3770 were monomorphic among all the clones. The Jaccard's similarity coefficient revealed that out of 45 clones derived from different explants, 44 were grouped into a single large cluster alongwith the mother plant with a similarity coefficient value of 1.00, whereas one clone (C38) remained ungrouped. The clones derived from capitulum and shoot tip explants did not show any genetic variation, whereas, one of the leaf-derived clones exhibited some degree of variation.     

Growth,feed efficiency and blood profile of buffalo calves consuming high levels of fluoride

Twelve male buffalo calves of 10 to 12 months of age were divided into 3 groups of four each. They were fed wheat straw+concentrate mixture +3 Kg greens. The chemical composition of the diet was same in all the three groups except fluoride which was added (as NaF) in concentrate mixture of group B and C to make the final fluoride concentration 30 ppm and 60 ppm respectively. The animals were kept on scheduled diet for a period of 90 days. Body weights were recorded at the start of the experiment and at fortnightly interval thereafter. Analysis of data revealed that the dry matter intake decreased non significantly in group B and C as compared to control group. A significant decrease in serum calcium and a significant increase in phosphorus concentration were observed in group C animals. A significant increase was observed in alkaline phosphatase activity in group C animals. A non significant decrease was observed in T₄ values in group C animals. On the basis of these results it could be concluded that fluoride in the diet of buffalo calves @ 30 ppm is a safe level whereas 60 ppm has affected the blood metabolites.     

Isotope-edited nuclear magnetic resonance: novel methodologies for investigating metabolism

While the use of NMR and stable isotopes in metabolism studies is hardly new, it is only recently that isotope-edited NMR spectroscopy has been applied in kinetic studies of glyphosate metabolism of soil microbes. NMR can detect multiple species simultaneously and non-destructively, yielding valuable information on structural identification of metabolites. T riple R esonance I sotope ED ited spectroscopy (TRIED), [²H]NMR, and [²H–¹³C] INEPT (I nsensitive N ucleus E nhancement through P olarization T ransfer) are three isotope-edited techniques which have been used in combination to examine the microbial degradation of glyphosate (N-phosphonomethylglycine). Using ¹³C- and ¹⁵N-labeled glyphosate, TRIED can detect multiple metabolites in crude matrices at submicrogram levels, an improvement over earlier techniques where milligrams were needed. It can detect 500 nanograms of ¹³C–¹⁵N-labeled compound in a crude sample (1 : 1400 mass ratio), only a few hours work being required. [²H]NMR and [²H–¹³C]INEPT were also used as complementary techniques to further examine metabolites whose ¹³C–¹⁵N bond has been cleaved. The three-isotope-edited methods produced results consistent with both radioactivity and HPLC analyses. Accordingly, we are able to detect minute levels of metabolites in the presence of complex mixtures, minimizing the costs and time of sample purification. ©1999 Society of Chemical Industry     

A comparative study on certain enzymes of the granulocyte from different ruminant species

In the present study the level of enzyme hydrolases (alkaline phosphatase, myeloperoxidase, elastase, arginase, lysozyme and β-galactosidase) of polymorphonuclear cell (PMN) granules in different ruminant species and their release in response to activation was studied. Buffalo PMN alkaline phosphatase activity was higher (P < 0.01) than in PMNs of cattle and goats. Interestingly, myeloperoxidase was higher in cattle PMNs and least in goat PMNs (P < 0.01), a similar pattern was observed in the distribution of enzyme arginase. As far as lysozyme is concerned, its activity was significantly higher (P < 0.01) in PMNs of buffaloes than in the case of cattle and goat PMNs. On activation, these cells released MPO and elastase, in all the species studied, while lysozyme was secreted only in buffalo PMN cells. Activity of certain enzymes related to oxidant defence systems such as glutathione peroxidase and glutathione reductase were higher in cattle and goats compared to that in buffaloes. These observations are likely to have bearing on immunodefense roles played by PMNs and reflected differences among the ruminant species studied.