Assessment of Awareness about Antibiotic Resistance and Practices Followed by Veterinarians for Judicious Prescription of Antibiotics: An Exploratory Study in Eastern Haryana Region of India

Tropical Animal Health and Production - The rise of antibiotic resistance and its global outreach has recently become a subject of increasing importance in veterinary research with an objective to...     

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.     

Evaluation of highly sensitive indigenous milk ELISA kit with fecal culture, milk culture and fecal-PCR for the diagnosis of bovine Johne's disease (BJD) in India

Country lacks indigenous diagnostic kits against Johne's disease in animals. Indigenous ELISA and IS 900 PCR kits, originally developed for goats and sheep, have been adapted for screening of lactating cows. Multiple diagnostic tests were used to screen 26 lactating dairy cows against Johne's disease. Milk ELISA was evaluated with fecal culture, milk culture and fecal PCR. Of the 26 samples from lactating cows, 84.6, 96.1, 88.4 and 23.0% were positive in fecal culture, milk culture, m-ELISA and m-PCR, respectively. Comparatively milk sediment and milk fat culture detected 84.6 and 76.9% cows positive, respectively. Comparatively fecal culture and milk culture detected 84.6 and 96.1% cows positive, respectively. M-ELISA detected 11.5, 0.0, 11.5, 61.0 and 15.3%, cows as negative, suspected, low positive, positive and strong positive, respectively. There was good correlation between milk and fecal culture with m-ELISA. Three negative cows in m-ELISA were also detected in milk and fecal culture. Of the 26 decontaminated fecal samples, 23.0% cows were positive using specific IS 900 f-PCR. Comparative evaluation of m-ELISA with fecal and milk culture showed agreement in 80.7 and 84.6% cows, respectively. Sensitivity of m-ELISA with respect to fecal and culture was 90.9 and 95.6%, respectively. Comparative evaluation of four tests (milk culture, fecal culture, m-ELISA and f-PCR) showed that only 15.3% cows were detected in all the four tests. In three tests (fecal and milk culture and m-ELISA), 57.6% cows were detected positive. None of the cow was exclusively detected in f-PCR. Of the four diagnostic tests used milk culture was most sensitive (96.15%), followed by fecal culture (86.6%), m-ELISA (76.9%) and IS 900 PCR (23.0%) for the diagnosis of bovine Johne's disease (BJD). Milk ELISA detected only one cow extra, which was negative in milk culture. In view of the simplicity, rapidity and efficacy present milk ELISA kit employing soluble protoplasmic antigen from native Map 'Bison type' genotype of goat origin can be reliable for screening of bovine population against Johne's disease in India.     

Effect of 12-hour road transportation on physiological, immunological and haematological parameters in bulls housed at different space allowances

The effects of transporting Holstein Friesian bulls (n=72; bodyweight 403+/-3.5 kg) for 12h by road were examined. Adrenal, haematological and immune responses, body temperature and performance were recorded. The animals had been previously housed for 96 days at three space allowances (1.2, 2.7 or 4.2m(2) per bull). The bulls were allocated to one of two treatments: T (transport for 12h; n=16 per space allowance) and C (control; n=8 per space allowance). Basal cortisol plasma concentrations and interferon (IFN)-gamma production from cultured lymphocytes did not show any statistically significant difference (P>0.05) following the housing period. Removing bulls from their home pens and walking them to the pre-loading crush facility, loading onto the transporter, and unloading following the 12h road journey, significantly (P<0.001) increased plasma cortisol concentration. The bulls housed at 4.2m(2) had greater (P<0.05) plasma cortisol concentrations than bulls housed at 1.2m(2) at loading, unloading, or on return to the crush holding facility; those housed at 1.2m(2) had greater (P<0.05) plasma cortisol concentrations than bulls housed at 2.7 and 4.2m(2) in their home pens after transport. There was an increased (P<0.05) plasma cortisol response in the T than in the C bulls following adrenocorticotrophic hormone administration. Transport significantly reduced (P<0.05) IFN-gamma production, lymphocyte % and body weight and significantly increased (P<0.05) neutrophils, eosinophils, packed cell volume, red blood cell numbers and haemoglobin. In conclusion, housing bulls for 96 days in a range of space allowances did not affect basal cortisol response or immune function parameters. Whereas transport increased plasma cortisol and reduced the immune response in the short-term, the changes were transient and within normal physiological ranges, suggesting that 12h road transport had no adverse effect on welfare status over the longer term. Furthermore, transport of bulls housed at increased space allowance (4.2m(2)/bull) resulted in a greater plasma cortisol response, albeit still within normal physiological range.     

Adventitious rooting in shoot cuttings of Azadirachta indica and Pongamia pinnata

Adventitious rooting in shoot cuttings of neem (Azadirachta indica) and karanj (Pongamia pinnata) was investigated in different seasons. The influence of auxins and B-vitamins (neem only) was evaluated. Maximum rhizogenesis coincided with the emergence of new sprouts i.e. February (neem) and March (karanj). Considering both rooting percentage and root biomass, IBA was the most effective auxin tested. Compared to auxin free controls, IBA (1000 ppm in neem and 800 ppm in karanj) significantly increased adventitious rooting percentage (80 to 100 % respectively), the numbers of roots and root biomass for both species. In neem, B-vitamin treatment yielded 40–50% rooting for cuttings. This was significantly greater than rooting for controls but equivalent to IAA treatments. In neem, some of the cuttings treated with 2000–3000 ppm IBA had well developed root systems without any sprouts. In karanj, all auxin treatments promoted sprouting. The level of total soluble sugars in the rooting zone of neem and karanj was higher in non-rooted than rooted cuttings.     

Low level of polymorphism detected by SSR probes in bread wheat

In-gel hybridization patterns were studied in a set of nine diverse bread wheat ( Triticum aestivum L. em. Thell) genotypes using 23 simple sequence repeat (SSR) probes in combination with 14 different restriction enzymes. Multilocus fingerprints due to SSR probes, shown earlier to be characteristic of a majority of plant genomes, were not obtained and only a very low level of polymorphism was detected when using as many as 142 probe-enzyme combinations. The hybridization of a prominent solitary high molecular weight fragment (> 23 kb) with a number of SSR probes suggested the presence of these SSRs (microsatellites) within the long stretches of repeated DNA sequences. This indicates that the genome of bread wheat differs from that of other plants in the organization and distribution of SSRs and that SSR probes detect very little polymorphism.     

Immobilization of cadmium and zinc in soil by Al-montmorillonite and gravel sludge

We investigated the potential of montmorillonite, Al-montmorillonite and gravel sludge to immobilize polluting heavy metals in agricultural soil. Batch experiments showed that both Al-montmorillonite and montmorillonite immobilized zinc and cadmium. Zinc was bound specifically on Al-montmorillonite and became increasingly incorporated into the interlayer hydroxy-Al polymer, whereas there was no specific sorption on montmorillonite. Cadmium was bound on montmorillonite and Al-montmorillonite unspecifically by cation exchange, but there was no incorporation into the lattice. In pot experiments montmorillonite, Al-montmorillonite, or gravel sludge were added to a soil contaminated with zinc and cadmium. Increasing doses of these agents decreased the concentrations of NaNO₃-extractable zinc and cadmium. Aluminium-montmorillonite and gravel sludge were more efficient than montmorillonite in immobilizing both zinc and cadmium. Remobilization tests at pH between 4 and 5.5 showed that cadmium and zinc desorbed more easily from montmorillonite than from Al-montmorillonite. Gravel sludge application increased the buffer capacity of the contaminated soil substantially. The binding agents decreased zinc concentrations in red clover (Trifolium pratense), and gravel sludge also reduced the cadmium concentrations.     

Processing of maize germ oil cake into edible food grade meal and evaluation of its protein quality

The commercial oil cake produced during expeller pressing of maize germ, was extracted with n-hexane and 80 percent ethanol followed by seiving to remove undesirable materials. In defatted maize germ oil cake (DMGOC): protein, starch, fat, crude fiber (CF) and ash were respectively 24.69, 36.55, 5.68, 7.56 and 3.90 percent and they decrease after processing except ash, which increased slightly. It contains better quality protein having only 3 percent zein and 47 percent albumin. Its amino acids like lysine and tryptophan and biological value (BV) were higher than that of whole maize grain, and was comparable with that of the amino acid requirement of preschool children and casein diets both. Its digestible energy (DE) was lower compared with whole maize grain as well as the casein diets. After processing albumin, globulin and zein decreased whereas glutelin and the residual fraction increased. Not much differences were observed in chemical composition and different amino acids, in vitro protein digestibility (IVPD), true digestibility (TD), BV and DE improved after processing.     

Organ culture of chicken bursa as a model to study the pathogenicity of infectious bursal disease virus isolates.

In vitro study with chicken bursal organ culture was attempted to assess the pathogenicity of locally isolated infectious bursal disease virus (IBDV) initially isolated from the bursa of naturally infected birds. In bursal organ culture, lymphoblastic transformation was noticed as early as 24 hr postinoculation and reached maximum at 72 hr postinoculation. The other microscopic changes were increased number of macrophages and formation of plasma cells. The IBDV antigen was detected 24 hr onward by coagglutination test with antibody coated Staphylococcus aureus strain Cowan I. On the basis of lesion score, the three isolates of IBDV (A, B, and C) were graded as virulent (B isolate) and moderately virulent (A and C isolates). A similar pattern of pathogenicity was also observed in the in vivo pathogenicity studies in chicken based on bursa: body weight ratio and histopathologic lesion score. The bursal organ culture thus provides a useful experimental model to differentiate the IBDV isolates on the basis of their virulence.     

Inheritance and allelic relationship among gene(s) for blast resistance in pearl millet [Pennisetum glaucum (L.) R. Br.]

Six blast‐resistant pearl millet genotypes, ICMB 93333, ICMB 97222, ICMR 06444, ICMR 06222, ICMR 11003 and IP 21187‐P1, were crossed with two susceptible genotypes, ICMB 95444 and ICMB 89111 to generate F₁s, F₂s and backcrosses, BC₁P₁ (susceptible parent × F₁) and BC₁P₂ (resistant parent × F₁) for inheritance study. The resistant genotypes were crossed among themselves in half diallel to generate F₁s and F₂s for test of allelism. The F₁, F₂ and backcross generations, and their parents were screened in a glasshouse against Magnaporthe grisea isolates Pg 45 and Pg 53. The reaction of the F₁s, segregation pattern of F₂s and BC₁P₁ derived from crosses involving two susceptible parents and six resistant parents revealed the presence of single dominant gene governing resistance in the resistant genotypes. No segregation for blast reaction was observed in the F₂s derived from the crosses of resistant × resistant parents. The resistance reaction of these F₂s indicated that single dominant gene conferring resistance in the six genotypes is allelic, that is same gene imparts blast resistance in these genotypes to M. grisea isolates.