Purification of fusaproliferin from cultures of Fusarium subglutinans by preparative high-performance liquid chromatography

Thirty-six Fusarium strains were grown on cracked yellow corn and evaluated for optimum fusaproliferin production, with Fusarium subglutinans E-1583 producing the highest levels (1600 microg/g). Three solvent systems were tested for extracting fusaproliferin from the cultures of F. subglutinans E-1583. Methanol gave the highest fusaproliferin recovery, followed by methanol/1% aqueous NaCl (55:45, v/v) and acetonitrile/methanol/H(2)O (16:3:1, v/v/v). Hexane partitioning was effective in removing many impurities from the crude fusaproliferin extracts prior to the liquid chromatography step. Fusaproliferin samples were further purified by high-performance liquid chromatography (HPLC) with a C18 preparatory column using a mobile phase of acetonitrile/H(2)O (80:20, v/v). The purity of the fusaproliferin was verified by analytical HPLC, GC/MS, (1)H NMR spectroscopy, and electrospray ionization (ESI) MS. The isolated fusaproliferin was shown to be free of impurities and can be used as a standard for routine analysis. Fusaproliferin was shown to be temperature-sensitive when samples were stored at room temperature (20-24 degrees C) for more than several days. After 30 days at 4 degrees C, approximately 8% of the fusaproliferin had been transformed to deacetyl-fusaproliferin; however, samples stored at -20 degrees C for 1 year contained only trace amounts of the deacetylated form.     

Diofenolan: a novel insect growth regulator in common citrus butterfly, <Emphasis Type="Italic">Papilio demoleus</Emphasis>

Penultimate and last instar larvae of the common citrus swallowtail butterfly, Papilio demoleus L. (Papilionidae: Lepidoptera), were treated with different doses of a novel juvenoid, diofenolan. Several deformities were observed as a result of topical administration of diofenolan which include delay in larval–larval and larval–pupal ecdysis, ecdysial failure, mortality, severe reduction in pupation, deformed pupae and complete inhibition of adult emergence. The juvenoid diofenolan severely hampers the normal growth, development and metamorphosis of P. demoleus and can be incorporated into integrated pest management programs for successful control of this important pest of citrus all over the world.     

Assessment of Atmospheric Arsenic Level in Airborne Dust Particulates of an Urban City of Central India

An assessment of arsenic contamination in Raipur city (21°14′N, 18°38′E) of Chhattisgarh in the central part of India is reported here, for a monitoring period between November 1996 to June 1997, in airborne dust particulates. The concentration level of As were higher in the industrial site, followed by heavy traffic as compared to other sites. The monthly atmospheric arsenic deposition, in μg As per g of dust fall, of 6 sites are in the range of 0.100(μ0.020)–4.00(μ0.020); site no. 1 industrial area, 0.100(μ0.020)–0.320(μ0.020); site no. 2 residential area, 0.044(μ0.070)–0.337(μ0.030); site no. 3 commercial area, 0.093(μ0.068)–1.870(μ0.020); site no. 4 residential area, 0.111(μ0.020)–1.912(μ0.010); site no. 5 residential area and 0.068(μ0.040)–3.037(μ0.060); site no. 6 heavy traffic area. The total annual flux of As in the fall-out at different zones is in the range 0.033–1.12 kg km^-2 yr^-1. The month wise collection and analysis of dust fall out rate between 3.0(μ0.10)–91.3(μ1.4) mt (metric tonnes) km^-2 month^-1 were observed at all 6 sampling sites. Anthropogenic and environmental factors play important roles in the contribution of arsenic in airborne particulate matters.     

Species affinities between Cajanus cajan and some Atylosia species based on esterase isoenzymes

Summary Esterase isozymes were studied in seed extracts of Cajanus cajan and six Atylosia species by polyacrylamide gel electrophoresis and isoelectrofocusing. The isozyme patterns were stable and accession specific. Within the accessions of the Atylosia species, A. albicans and A. scarabaeoides showed three common bands indicating that they are more closely related to each other than to the other species. Of the accessions of Atylosia only A. cajanifolia shares the esterase isozyme of C. cajan and hence seems to be the closest wild relative of C. cajan.     

Potential of a novel antibiotic, 2-methylheptyl isonicotinate, as a biocontrol agent against fusarial wilt of crucifers

Screening for newer bioactive compounds from microbial metabolites resulted in the isolation of a novel antibiotic from the culture filtrate, Streptomyces sp 201, of a soil. The bioactive compound, with antifungal and antibacterial activity, was identified as 2-methylheptyl isonicotinate. The antifungal activity of live culture, culture broth and the isolated bioactive compound showed marked inhibition against dominant soil-borne phytopathogens such as Fusarium oxysporum Schlect, F moniliforme Sheldon, F semitectum Berkeley & Ravenel, F solani (Martius) Sacc and Rhizoctonia solani Kuehn. The compound had no effect on seed germination and seedling development as displayed by root and stem growth of the test plant species. In pot experiments with seedlings of cruciferous plants such as Raphanus sativus L (radish), Brassica campestris L (yellow mustard), Brassica oleracea var botrytis L (cauliflower), the antibiotic compound showed promising protective activity of 92% when seeds of the test plants were treated at a dose of 50 micrograms ml-1 prior to sowing. Seed treatment with a spore suspension (3 x 10(8) spores ml-1) of the Streptomyces sp 201 displayed protective activity in the range of 56-60%. Seeds coated with 2.5% methyl cellulose-amended spores of the antagonist showed protective activity in the range of 64-72%. Further, seed treatment with the culture filtrate of the antagonist also showed promising protective activity in the range of 64-84%.     

Biological Control of Late Leaf Spot of Peanut (Arachis hypogaea) with Chitinolytic Bacteria

ABSTRACT Late leaf spot (LLS), caused by Phaeoisariopsis personata, is a foliar disease of groundnut or peanut (Arachis hypogaea) with high economic and global importance. Antifungal and chitinolytic Bacillus circulans GRS 243 and Serratia marcescens GPS 5, selected among a collection of 393 peanut-associated bacteria, were applied as a prophylactic foliar spray and tested for control of LLS. Chitin-supplemented application of B. circulans GRS 243 and S. marcescens GPS 5 resulted in improved biological control of LLS disease. Supplementation of bacterial cells with 1% (wt/vol) colloidal chitin reduced lesion frequency by 60% compared with application of bacterial cells alone, in the greenhouse. Chitinsupplemented application of GRS 243 and GPS 5 also resulted in improved and stable control of LLS in a repeated field experiment and increased the pod yields by 62 and 75%, respectively, compared with the control. Chitin-supplemented application of GPS 5 was tested in six onfarm trials, and the increase in pod yields was up to 48% in kharif (rainy season). A 55-kDa chitinase was purified from the cell-free culture filtrate of GPS 5 by affinity chromatography and gel filtration. Purified chitinase of S. marcescens GPS 5 (specific activity 120 units) inhibited the in vitro germination of P. personata conidia, lysed the conidia, and effectively controlled LLS in greenhouse tests, indicating the importance of chitinolysis in biological control of LLS disease by GPS 5.     

An outbreak of pine lappet moth, Kunugia latipennis, in mid-altitude hills of Meghalaya state, India

The pine lappet moth, Kunugia latipennis Walker (Lepidoptera: Lasiocampidae), is the major insect pest of pines found in the northeastern hilly region of India. Recently, an outbreak of lappet moths was observed in the mid-altitude hills of Meghalaya during May?CJune 2011. In this report, attempts are made to describe a recent outbreak of K. latipennis in the region. Females were found to be highly fecund and have the potential to cause severe damage to the pine forest. Incubation and pupal period were found to be 8?±?0.2?days and 16.1?±?0.3?days, respectively. However, many natural regulatory factors were observed during the course of time, which had substantial impact on their survival: these include mainly extreme weather fluctuations and natural enemies. Moths were found to be positively phototrophic; therefore, light traps could be the best option to manage the outbreak.     

Diagnosis of paratuberculosis in goats by cell mediated immune response,conventional and molecular diagnostic techniques

In the present study efficacy of single intradermal Johnin test, acid fast staining of faecal smear and IS 900 faecal polymerase chain reaction tests was evaluated in 200 goats for detection of Mycobacterium avium subsp paratuberculosis. Two hundred goats comprising 150 goats from an organised farm in Trichur district and 50 goats reared under field condition at farmers premise from Malappuram district of Kerala state formed the study population. Faecal smear from all the 200 goats was stained by Ziehl–Neelsen acid fast stain and faecal polymerase chain reaction (PCR) specific for M. avium subsp paratuberculosis (MAP); IS 900 was performed on all samples. All the animals were subjected to single intradermal Johnin test. Out of 200 goats screened for paratuberculosis, six goats (3%), 11 goats (5.5%) and 42 goats (21%) were found positive by Ziehl–Neelsen acid fast staining of faecal smear, single intradermal Johnin test and IS 900 PCR respectively. Results of the present study indicate that amplification of IS 900 insertion element was the most specific and sensitive diagnostic detection method. Single intradermal Johnin test and Ziehl–Neelsen acid fast staining did not show any significant difference.