Molecular detection of important abortion-causing microorganisms in preputial swab of breeding bucks using PCR-based assays

Infectious diseases and aetiological agents related to female reproductive systems were extensively covered compared to its male counterpart. There needs a proper study to bridge this gap, where microflora and infectious agents of both male and female reproductive are mutually intelligible. With this study, we aimed to evaluate the microbial contamination of the preputial cavity and also screened for abortion-causing agents which are zoonotic as well. In goats, such types of abortions are caused by Brucella melitensis, Chlamydophila, Campylobacter and Coxiella etc. One of the major sources of contamination of semen is the preputial cavity, which is exposed to the external environment leading to spread of infection into the female via semen straws or by natural service. In the current study, good quality bucks (n = 32, Barbari = 12, Jamunapari = 10, Jakhrana = 10) which were routinely used for semen collection were screened for their preputial swabs, for the presence of the above pathogens. For detection of Brucella melitensis, OMP31 based TaqMan® probe real-time PCR assay was used, and for Chlamydia, 16srRNA gene based SYBR® green real-time PCR assay was employed for detection of Chlamydophila abortus. While for Campylobacter spp. and Coxiella burnetii, 16srRNA gene based conventional PCR and Trans-PCR were used, respectively. In the current study, of the screened preputial swabs, none of them showed positive for Brucella and Coxiella, but of the screened 32 samples 17 showed positive for Chlamydia (53.13%) and two (6.25%) showed positive for Campylobacter spp. The current study emphasizes on the farms and laboratories which were regularly involved in screening of brucellosis also often overlook the other potential non-brucella pathogens, causing abortions eventually incurring severe economic losses to the goat keepers.     

Effect of coconut palm wine (Toddy) on carbohydrate metabolism in pregnant rats and fetuses

The objective of this study was to determine the effects of an alcoholic beverage (Toddy) and the equivalent quantity of ethanol on carbohydrate metabolism in utero. Female rats were exposed to Toddy from coconut palm (24.5 ml/kg body weight/day) and ethanol (0.52 ml/kg body weight/day) for 15 days before conception and throughout gestation. On the 19th day of gestation, hypoglycemia was seen in both the treated groups, but it was more in the Toddy-treated group. Synthesis of glycogen was elevated on exposure to ethanol/Toddy but its degradation was enhanced only in alcohol-exposed rats. Key enzymes of citric acid cycle and gluconeogenesis were inhibited on administration of both alcohol and Toddy. Activity of glycolytic enzymes were increased. Toddy seemed to potentiate the toxicity induced by alcohol, indicating the additive effects of congeners.     

Molecular characterization and variability analysis of <Emphasis Type="Italic">Apple scar skin viroid</Emphasis> in India

Apple scar skin viroid (ASSVd) infection is a major limitation to apple fruit quality and causes huge economic losses. In surveys of apple orchards in the northern Indian state of Himachal Pradesh, fruits with dappling symptoms were noticed. ASSVd was detected from these fruits and molecularly characterized. Ten clones from three isolates were sequenced, of which seven were new sequence variants of ASSVd. The clones had significant sequence variability (94–100%) with each other. Variability was more common in the pathogenic domain of the viroid genome. Four of the clones were 330 nucleotides (nt) long, and the other six had an additional nucleotide. Phylogenetic analysis showed close affinity of the present isolates with some Chinese and Korean isolates. The study reports seven new variants of ASSVd and also provides the first molecular evidence of viroid infection (ASSVd) in apple in India.     

Hypoglycemic activity of Nymphaea stellata leaves ethanolic extract in alloxan induced diabetic rats

The ethanolic extract of leaves of Nymphaea stellata given by oral route to diabetic rats at dose of 100 and 200 mg/kg/day for seven days reduced significantly by 31.6 and 42.6 % the plasma glucose level increased by intraperitoneal injection of 120 mg/day of alloxan. Moreover, the treatment significantly affected the plasma level of cholesterol and triglyceride.     

Antibacterial activity of Mappia foetida leaves and stem

Successive petroleum ether, chloroform and methanol extracts of Mappia foetida leaves and stem were tested for their antibacterial activity. The methanol fractions were found to be most effective against all the tested organisms.     

Effect of dihydrodillapiole on pyrethroid resistance associated esterase inhibition in an Indian population of Spodoptera litura (Fabricius)

Resistance in Spodoptera litura (Fabricius) has been attributed to enhanced detoxification of insecticides by increased levels of esterases, oxidases and/or glutathione S-transferases. Enzyme inhibiting insecticide synergists can be employed to counter increased levels of such enzymes in S. litura. Dihydrodillapiole induced synergism of pyrethroid toxicity was examined in the laboratory-reared third instar larval population of S. litura collected in Delhi (susceptible), and Guntur (resistant) region of Andhra Pradesh, India. The Guntur population was found to be 7.04 and 10.19 times resistant to cypermethrin and lambdacyhalothrin, respectively. The activity of cypermethrin, lambdacyhalothrin and profenophos against susceptible and resistance populations of S. litura, was gradually increased when used along with a plant-derived insecticide synergist dihydrodillapiole. The α-naphthyl acetate hydrolysable esterase activity in Delhi population was less as compared to the Guntur population. Resistance associated esterases in Delhi population were inhibited by pre-treatment with dihydrodillapiole. The esterase level in insect was instantly reduced initially, sustained for about 3 h and equilibrated at 4 h post treatment. The esterase activity of Guntur population was increased to 1.28 μmoles/mg/min at 2 h post treatment and subsequently reduced to lower than 0.70 μmoles at 4-12 h post treatment. The variation in esterase activity is suggestive of its homeostatic regulation in test populations. Dihydrodillapiole thus caused significant reduction of resistance in S. litura to cypermethrin, lambda cyhalothrin and profenophos.     

Efficacy of 17 α‐methyl testosterone and letrozole on sex reversal of protogynous grouper,Epinephelus tauvina (Forskal, 1775) and spawning performance of sex‐reversed males

This study aimed to test the efficacy of 17 α‐methyl testosterone (17 α‐MT) alone and in combination with letrozole, an aromatase inhibitor, for the induction of sex reversal in protogynous greasy grouper, Epinephelus tauvina. Further, the long‐lasting effects of these treatments and spawning performance of sex‐reversed males were also investigated. Greasy grouper with oocytes in the perinucleolus stage were implanted with 5 mg 17 α‐MT kg^?1 body weight (T₁), 5 mg 17 α‐MT and 0.2 mg letrozole kg^?1 body weight (T₂) and 5 mg 17 α‐MT with 0.4 mg letrozole kg^?1 body weight (T₃) and no androgens/enzyme inhibitor implanted (C). The 17 α‐MT alone and in combination of letrozole‐induced sex reversal in greasy grouper, whereas untreated control fish (C) showed normal ovarian development. However, T₂ and T₃ group showed 100% sex reversal and completion of spermatogenesis up to functional male phase in 2 and 3 months, respectively, whereas T₁ group resulted in only 66.67% functional male with motile spermatozoa after 4 months. Sex‐reversed males successfully fertilized the eggs during induced spawning. There were significant differences on fertilization and hatching rates between T₂ group (79.00 ± 4.36%; 77.67 ± 2.87%, respectively) and T₁ group (57.67 ± 3.17%; 63.87 ± 2.91%, respectively). The result suggested that 17 α‐MT (5.0 mg kg^?1 BW) in combination with letrozole (0.2 mg kg^?1 BW) has the potential to produce 100% sex‐reversed male in short period in greasy grouper, which might greatly help in seed production of greasy grouper.     

Effect of Spirulina fusiformis on cyclophosphamide and mitomycin-C induced genotoxicity and oxidative stress in mice

Spirulina fusiformis was tested for its possible in vivo protective effects against cyclophosphamide (CP) and mitomycin-C (MMC) induced genotoxicity and oxidative stress in mice. Pre-treatment with S. fusiformis (250, 500 and 1000 mg kg(-1), p.o., daily for 5 days) significantly reduced the chromosomal damage and lipid peroxidation with concomitant changes in antioxidants and detoxification systems. All the three tested doses were effective in exerting a protective effect against CP and MMC.     

Assessment of nifH diversity in rhizobial isolates of different origin and the role of antioxidant in respiratory protection

Rhizobia diversity is considered as one of the most useful resources for bioprospecting due to their symbiotic nitrogen-fixing ability with members of Leguminosae. The highly conserved nature of the nitrogenase reductase gene (nifH) makes it an ideal molecular tool to determine the potential for biological nitrogen fixation in any environment. In the present investigation, 250 rhizobial strains were isolated from legumes belonging to different geographical locations of Chhattisgarh, India. Genetic diversity of the nitrogenfixing bacterial community was analyzed using the nifH gene-specific primer. The polymorphism was found among the nitrogen-fixing population of different sources and origin but not in same source of rhizobia. Further, the symbiotic plasmid DNA was characterized on the basis of size and copy number of plasmids. The plasmid number varying from one to three in different rhizobial isolates had a size greater than 23 kb, while in some rhizobial isolates plasmids were absent. In addition, to examine the role of ascorbate in respiratory protection, the clear black spot margin of ascorbate was observed in the endodermis region of the nodule whereas scarcely dispersed in the infected region. Therefore, our findings demonstrated that knowing the rhizobial nifH gene diversity along with copy number of the plasmid is important for strain identification, deciding its fertility, productivity standards, and potential of biological nitrogen fixation across the geographical region.     

DDT and HCH Residues in Basmati Rice (Oryza sativa) Cultivated in Dehradun (India)

Organochlorine pesticides were used earlier for agricultureproduction. Their residues may still be present in soil and mayaccumulate in food crops, posing potential health problems to consumers. DDT, HCH, their isomers and metabolites were analyzedin samples of soil and rice plants collected from ten differentvillages of a well-known Basmati rice growing area in Dehradun.Residues of both pesticides were found in all samples ofsoil and different parts of rice plants except for a few grainsamples. Maximum residue was observed in husk and minimum ingrains. The average concentration of DDT in soil ranged from0.013 to 0.238 ppm. p,p′-DDE was the major metabolite (>63%). Theaverage concentration of DDT in rice grain varied from 0.002 to 0.040 ppm. o,p′-DDT was the main isomer (>93%). Theaverage concentration of HCH in soil ranged from 0.122 to 0.638 ppm. β-HCH was the predominant (43%) isomerfollowed by α-HCH (21%). The average HCH concentrationin rice grain ranged between 0.013 and 0.113 ppm. All four isomers were present in grains. The levels of DDT and CHCin grains were similar in magnitude as those from differentIndian states, but well below the maximum residue limit of 0.1 ppm for DDT and 0.05 ppm for HCH prescribed by the Government ofIndia and WHO/FAO. As such, the pesticide residue levels in thisexport commodity are not of hazardous nature.