An efficient method for the purification and characterization of nematicidal azadirachtins A,B, and H,using MPLC and ESIMS

Azadirachtin A enriched concentrate containing 60% active ingredient (a.i.) was prepared from the methanolic extract of the de-fatted neem (Azadirachta indica A. Juss) seed kernels. Azadirachtins A, B, and H, the three major bioactive constituents of neem seed kernel, were purified from this methanolic concentrate by employing reverse phase medium-pressure liquid chromatography (MPLC), using methanol-water solvent system as an eluant. The three pure azadirachtin congeners thus obtained were characterized by their unique mass spectral fragmentation, using electrospray probe in positive ion mode (ESI). All three azadirachtins exhibited nematicidal and antifungal activities. Azadirachtin B was the most effective against the reniform nematode Rotylenchulus reniformis (EC(50) 96.6 ppm), followed by Azadirachtin A (119.1 ppm) and H (141.2 ppm). At 200-ppm concentration, the test compounds caused 50-65% mortality of Caenorhabditis elegans nematode. Azadirachtin H showed the highest activity against the phytophagous fungi Rhizoctonia solani (EC(50) 63.7 ppm) and Sclerotium rolfsii (EC(50) 43.9 ppm), followed by B and A. The isolation of pure azadirachtins A, B, and H directly by MPLC purification from its concentrate and their characterization by ESIMS are unique and less time-consuming.     

Effect of gamma-irradiation on the lipid profile of nutmeg (Myristica fragrans Houtt.)

The effect of gamma-irradiation on the lipid constituents of nutmeg (Myristica fragrans) was examined at radiation doses between 2.5 and 10 kGy. The fatty acid composition of the triacylglycerol, the major lipid component, was found to be made up of myristic (90%), palmitic (6%), lauric (3%), petroselinic (0.13%), and stearic acids (0.5%) as determined by gas chromatography-mass spectrometry. A dose-dependent decrease in the triacylglycerol content and a concomitant increase in free fatty acids characterized the lipid profile of the irradiated spice. This suggested a breakdown of acylglycerols during radiation processing, resulting in the release of free fatty acids. These changes were found to be significant at doses above 5 kGy. The impact of the above changes on the flavor of the spice is discussed. These studies suggest that radiation processing of nutmeg should be limited to a dose of 5 kGy.     

Subcutaneous pharmacokinetics and dosage regimen of cefotaxime in buffalo calves (Bubalus bubalis)

The pharmacokinetics and dosage regimen of cefotaxime following its single subcutaneous administration (10 mg/kg) were investigated in buffalo calves. Plasma and urine samples were collected over 10 and 24 h post administration, respectively. Cefotaxime in plasma and urine was estimated by microbiological assay technique using E. coli as test organism. The pharmacokinetic profiles fitted one-compartment open model. The peak plasma levels of cefotaxime were 6.48 ± 0.52 µg/ml at 30 min and the drug was detected upto 10 h. The absorption half-life and elimination half-life were 0.173 ± 0.033 h and 1.77 ± 0.02 h, respectively. The apparent volume of distribution and total body clearance were 1.17 ± 0.10 l/kg and 0.45 ± 0.03 l/kg/h, respectively. The urinary excretion of cefotaxime in 24 h, was 5.36 ± 1.19 percent of total administrated dose. A satisfactory subcutaneous dosage regimen for cefotaxime in buffalo calves would be 13 mg/kg repeated at 12 h intervals.     

Papaya yellow leaf curl virus: A newly identified begomovirus infecting Carica papaya L. from the Indian Subcontinent

India is one of the world’s largest producers of papaya. Viruses, mainly begomoviruses and potyviruses, cause a significant loss in papaya production. The study described here has identified a new species of begomovirus and a new species of betasatellite infecting Carica papaya in India. The sequences of the begomovirus and betasatellite show 90.03% nucleotide sequence identity to an isolate of Radish leaf curl virus and 92.25% identity to an isolate of Tomato leaf curl betasatellite, respectively. Maximum likelihood phylogenetic tree of begomovirus sequence of isolate DP2 (KX353622) showed distant relationships with previously characterised begomoviruses. Recombination analysis proposed six recombination breakpoints in begomovirus genome with other geographical begomovirus isolates.     

Achyranthes aspera (Prickly chaff flower) leaves- and seeds-supplemented diets regulate growth,innate immunity,and oxidative stress in Aeromonas hydrophila-challenged Labeo rohita

ABSTRACT

The immunostimulatory and disease-resistance properties of Achyranthes aspera were evaluated in rohu (Labeo rohita) challenged with Aeromonas hydrophila. Experimental diets were enriched with leaves at 0.25% (D1) and 0.5% (D2) and seeds at 0.5% (D3); the control diet (D4) was without any enrichment. Rohu (2.02 ± 0.23 g) were cultured for 75 days and then challenged with bacteria. The highest average weight was observed in the D3 diet-fed fish. The cumulative mortality rates were 70%, 60%, 40%, and 30% in the D4, D1, D2, and D3 diets fed to rohu respectively. Enriched diets significantly increased myeloperoxidase, nitric oxide synthase, and serum lysozyme levels and decreased malondialdehyde and carbonyl protein content. Expressions of lysozyme C and lysozyme G were significantly (P < .05) higher in the D3 diet-fed fish. In the kidney, IL-1β and TLR 4 were up-regulated in enriched-diet-fed fish. Supplementation of seeds and leaves at 0.5% showed a positive impact in fish.     

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...     

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.     

Wild relatives of sorghum as sources of resistance to sorghum shoot fly, Atherigona soccata

The levels of resistance to shoot fly, Atherigona soccata in sorghum germplasm are low to moderate and therefore, we evaluated 17 wild relatives of sorghum under field and greenhouse conditions as an alternate source of genes for resistance to this pest. Thirty-two accessions belonging to Parasorghum , Stiposorghum and Heterosorghum did not suffer any shoot fly damage under multi-choice conditions in the field, while one accession each of Heterosorghum ( Sorghum laxiflorum ) and Chaetosorghum ( S. macrospermum ) suffered very low shoot fly damage. Accessions belonging to S. exstans (TRC 243601), S. stipoideum (TRC 243399) and S. matarankense (TRC 243576) showed absolute non-preference for oviposition under no-choice conditions. Accessions belonging to Heterosorghum , Parasorghum and Stiposorghum were preferred for oviposition, but suffered low deadheart formation. Manual infestation of seedlings with shoot fly eggs did not result in deadheart formation in some of the accessions belonging to S. exstans (TRC 243601), S. stipoideum (TRC 243399), S. matarankense (TRC 243576) and S. purpureosericeum (IS 18944). Larval mortality was recorded in main stems of the Parasorghums . Within section Sorghum , accessions belonging to S. bicolor ssp. verticilliflorum were highly susceptible to shoot fly, as were those of S. halepense . However, a few accessions such as IS 18226 (race arundinaceum ) and IS 14212 ( S. halepense ) resulted in reduced survival and fecundity. Wild relatives of sorghum exhibited very high levels of antibiosis to A. soccata , while only low levels of antibiosis have been observed in the cultivated germplasm. Therefore, wild relatives with different mechanisms of resistance can be used as a source of alternate genes to increase the levels and diversify the basis of resistance to shoot fly, A. soccata .