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.     

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.     

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.     

Characterization of the long-term immune response to vaccination against Mycobacterium avium subsp. paratuberculosis in Danish dairy cows

Vaccination of cattle against Mycobacterium avium subsp. paratuberculosis (MAP) provides partial protection by delayed shedding of MAP and reduced numbers of clinically affected animals. The duration of vaccine induced immune response is not known. The primary objective of this study was therefore to characterize the long-term effect of whole-cell based vaccination against MAP on the immune response. A secondary objective was to evaluate whether immunodiagnosis of MAP and Mycobacterium bovis infections is affected by MAP vaccination. Two studies were performed: (1) A retrospective longitudinal study including 895 vaccinated and 2526 non-vaccinated dairy cows in 9 Danish dairy herds aiming at characterizing the long-term antibody-response to vaccination; and (2) a cross-sectional study of responses in the IFN-γ assay carried out in 140 vaccinated animals in two herds to evaluate the effect of vaccination on the cell-mediated immune response and to evaluate a possible interference with the diagnosis of M. bovis infections. The results showed that 37% of samples from vaccinated animals and 5% of samples from non-vaccinated animals, respectively, were test positive in the milk antibody ELISA. The prevalence of antibody responses of the vaccinated animals was relatively constant from 2 to 6 years of age, but decreased in older animals. Among the 140 vaccinated animals 88% tested positive with the IFN-γ test to johnin PPD and 50% responded to PPDb with IFN-γ production above a similar cut-off. Although Denmark is free of M. bovis, two of the vaccinated animals responded with higher IFN-γ levels when cultured with PPDb compared to PPDa. In conclusion, immunization with whole-cell MAP vaccines elicits both humoral and cell-mediated immune reactions, which may interfere with surveillance and diagnosis of both MAP and M. bovis infections using currently available tests.     

Longitudinal study comparing the dynamics of Clostridium difficile in conventional and antimicrobial free pigs at farm and slaughter

Clostridium difficile is the leading cause of nosocomial diarrhea in humans and a major cause of enteritis in neonatal piglets, foals and calves. The aim of this longitudinal study was to determine and compare the prevalence, antimicrobial susceptibility, and toxinotype profiles of C. difficile isolated from pigs and their environment in the indoor conventional and outdoor antimicrobial free (ABF) production systems. Ten conventional and eight ABF cohorts of 35 pigs each and their environment were sampled at different stages of production at farm and slaughter. C. difficile prevalence in pigs was highest at the farrowing stage in both conventional (34%, 120/350) and ABF (23%, 56/244) systems, and decreased with age. This reduction in C. difficile prevalence in pigs at later stages of production mirrored the decreased prevalence in the farm environment. At slaughter, C. difficile was isolated at a low frequency from the carcasses and processing environment in both production systems. All but three isolates were resistant to ciprofloxacin (99%, 505/508), while 1.0% (5/508) and 6.0% (23/508) of isolates exhibited resistance to tetracycline and erythromycin, respectively. Toxinotype V (tcdA(+)tcdB(+)) was the predominant strain identified in both systems (conventional: 94%, 376/401; ABF: 82%, 88/107), while the rest were toxinotype XIII (tcdA(+)tcdB(+)). To conclude, we isolated antimicrobial resistant C. difficile regardless of antimicrobial use on the farm. Based on the phenotypic and genotypic similarity of C. difficile isolated in this study, we conclude that the unique production practices employed in conventional and ABF production systems have no impact on the pathogen population.     

Assessment of biomass and net primary productivity of a dry tropical forest using geospatial technology

This study quantifies biomass, aboveground and belowground net productivity, along with additional environmental factors over a 2-3 year period in Barnawapara Sanctuary of Chhattisgarh, India through satellite remotesensing and GIS techniques. Ten sampling quadrates20×20, 5×5 and 1×1 m were randomly laid for overstorey (OS), understorey (US) and ground vegetation(GS), respectively. Girth of trees was measured at breast height and collar diameters of shrubs and herbs at 0.1 m height. Biomass was estimated using allometric regression equations and herb biomass by harvesting. Net primary productivity (NPP) was determined by Ssumming biomass increment and litter crop values. Aspect and slope influenced the vegetation types, biomass and NPP in different forests. Standing biomass and NPP varied from 18.6 to101.5 Mg ha^-1 and 5.3 to 12.7 Mg ha^-1 a^-1, respectively,in different forest types. The highest biomass was found in dense mixed forest, while net production recoded in Teak forests. Both were lowest in degraded mixed forests of different forest types. OS, US and GS contributed 90.4, 8.7and 0.7%, respectively, for the total mean standing biomass in different forests. This study developed spectral models for the estimation of biomass and NPP using Normalized Difference Vegetation Index and other vegetation indices.The study demonstrated the potential of geospatial tools for estimation of biomass and net productivity of dry tropical forest ecosystem.     

Lysimeter study on the use of biodrainage to control waterlogging and secondary salinization in (canal) irrigated arid/semi-arid environment

The study describes the capacity of trees to control the rise in water table and thus prevent the formation of waterlogged soils and development of secondary salinization in canal irrigated areas. It was conducted in RCC lysimeters of 1.2 m dia. and 2.5 m depth filled with sandy loam alluvial soil (Typic Ustochrept), with provisions to maintain water table depth at 1, 1.5 and 2 m from the surface and groundwater salinity at 0.4, 3, 6, 9 and 12 dS m-1. The amount of water biodrained by eucalyptus (Eucalyptus tereticornis) and bamboo (Bambusa arundinacea) at the given water table depths and groundwater salinity levels was monitored over four years by daily measuring the water needed for maintaining the water table. The trees continued to absorb and transpire water throughout the year, the capacity being more in summer and rainy than that was in the winter season. The eucalyptus plant could biodrain 2880, 5499, 5518 and 5148 mm of water in the first, second, third and fourth year of study period, from non-saline groundwater and a water table depth of 1.5 m. The amount of water biodrained was more at 1.5 m as compared to 1 and 2 m water table depths. The biodrainage capacity of trees was significantly affected by the salinity of the groundwater. However, even at salinity of 12 dS m-1, the eucalyptus plant biodrained 53% of that under non-saline conditions. It was calculated that biodrainage could control water table rises upto 1.95, 3.48, 3.76 and 3.64 m in first, second, third and fourth year, respectively. The secondary salinity developed in the root zone, upto 45 cm depth, did not exceed 4 dS m-1 even at water table depth of 1 m with salinity of 12 dS m-1. The volume of water biodrained by bamboo increased with time and could control water table rises upto 1.09, 1.86, 2.46 and 2.96 m in first, second, third and fourth year of growth, respectively.This study indicates that due to high transpiration capacity and an ability to extract water from deeper layers containing saline groundwater, the trees can control the rise in water table in irrigation command areas and prevent the formation of waterlogged and eventually the saline wastelands.     

Infectious agent detections in archived Sockeye salmon (Oncorhynchus nerka) samples from British Columbia,Canada (1985–94)

In response to concerns that novel infectious agents were introduced through the movement of eggs as Atlantic salmon aquaculture developed in British Columbia (BC), Canada, we estimated the prevalence of infectious agents in archived return‐migrating Sockeye salmon, from before and during aquaculture expansion in BC (1985–94). Of 45 infectious agents assessed through molecular assays in 652 samples, 23 (7 bacterial, 2 viral and 14 parasitic) were detected in liver tissue from six regions in BC. Prevalence ranged from 0.005 to 0.83 and varied significantly by region and year. Agent diversity ranged from 0 to 12 per fish (median 4), with the lowest diversity observed in fish from the Trans‐Boundary and Central Coast regions. Agents known to be endemic in Sockeye salmon in BC, including Flavobacterium psychrophilum, Infectious haematopoietic necrosis virus, Ceratonova shasta and Parvicapsula minibicornis, were commonly observed. Others, such as Kudoa thyrsites and Piscirikettsia salmonis, were also detected. Surprisingly, infectious agents described only recently in BC salmon, Ca. Branchiomonas cysticola, Parvicapsula pseudobranchicola and Paranucleospora theridion, were also detected, indicating their potential presence prior to the expansion of the aquaculture industry. In general, our data suggest that agent distributions may not have substantially changed because of the salmon aquaculture industry.     

Design standards for experimental and field studies to evaluate diagnostic accuracy of tests for infectious diseases in aquatic animals

Design and reporting quality of diagnostic accuracy studies (DAS) are important metrics for assessing utility of tests used in animal and human health. Following standards for designing DAS will assist in appropriate test selection for specific testing purposes and minimize the risk of reporting biased sensitivity and specificity estimates. To examine the benefits of recommending standards, design information from published DAS literature was assessed for 10 finfish, seven mollusc, nine crustacean and two amphibian diseases listed in the 2017 OIE Manual of Diagnostic Tests for Aquatic Animals. Of the 56 DAS identified, 41 were based on field testing, eight on experimental challenge studies and seven on both. Also, we adapted human and terrestrial‐animal standards and guidelines for DAS structure for use in aquatic animal diagnostic research. Through this process, we identified and addressed important metrics for consideration at the design phase: study purpose, targeted disease state, selection of appropriate samples and specimens, laboratory analytical methods, statistical methods and data interpretation. These recommended design standards for DAS are presented as a checklist including risk‐of‐failure points and actions to mitigate bias at each critical step. Adherence to standards when designing DAS will also facilitate future systematic review and meta‐analyses of DAS research literature.