Silymarin inhibits adipogenesis in the adipocytes in grass carp <Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis> in vitro and in vivo

In this study, two experiments were performed to explore the function of silymarin in adipogenesis in grass carp (Ctenopharyngodon idellus) using in vitro and in vivo models. In experiment 1, differentiated grass carp pre-adipocytes were treated with silymarin for 6 days. Treatment with 100 μg mL^?1silymarin (SM100 group) significantly reduced triglyceride accumulation at day 6. The adipogenic gene expression levels of PPARγ, C/EBPα, SREBP1c, FAS, SCD1, and LPL, and the protein expression level of PPARγ were significantly down-regulated in the SM100 group. Additionally, the SM100 group had significantly lower reactive oxygen species production and reduced glutathione contents compared with the control in vitro. In experiment 2, the juvenile grass carp (mean body weight= 27.4 ± 0.17 g) were fed six isonitrogenous and isocaloric diets in a factorial design containing 0, 100, or 200 mg kg^?1 silymarin (SM0, SM100, SM200) associated with either 4 or 8% lipid levels (low lipid, LL, and high lipid, HL, respectively) for 82 days. The results demonstrated that dietary silymarin supplementation significantly reduced the elevated intraperitoneal fat index in grass carp fed with high-lipid diets, and the gene expression of adipogenesis (PPARγ, FAS) when supplemented with dietary silymarin was notably lower than when no silymarin was supplemented under the high-lipid diets. Thus, our data suggest that silymarin suppressed lipid accumulation in grass carp both in vitro and in vivo, and the effect might be due to an influence on the expression of adipogenesis factors and ROS production partly associated with effects on antioxidant capability.     

Prevalence,molecular fingerprinting and drug resistance profile of enterovirulent <Emphasis Type="Italic">Escherichia coli</Emphasis> isolates from free-ranging yaks of Tawang district,Arunachal Pradesh,India

Of 273 samples (rectal swab) collected from free-ranging yaks of Tawang district, Arunachal Pradesh, 42 Shiga toxin-producing Escherichia coli (STEC), six enteropathogenic E. coli (EPEC) and 27 enterotoxigenic E. coli (ETEC) strains were isolated. All the STEC and EPEC strains were further investigated for respective stx variants (for STEC only) and additional putative virulence factors. The 27 ETEC strains were also screened for characteristic enterotoxin gene(s) and colonization factors. Occurrence of ETEC was significantly (p < 0.05) higher in the diarrheic yaks and yaks of less than 1 year of age. Majority of enterovirulent E. coli isolates were resistant to amikacin, azithromycin, chloramphenicol, colistin, doxycycline, furazolidone, nalidixic acid, nitrofurantoin, streptomycin and tetracycline. Dendrogram, constructed with molecular fingerprinting profiles obtained from RAPD (Randomly Amplified Polymorphic DNA) and ERIC (Enterobacterial Repetitive Intergenic Consensus) PCR, placed the isolates in different clusters irrespective of their serotypes, virulence gene and drug resistance pattern. Collectively, the study indicates that yaks, being a potential reservoir of multidrug resistant STEC and EPEC, may represent significant risk to public health in this region. Higher recovery of ETEC isolates from yaks with diarrhea points out that ETEC may be a major determinant for repeated occurrence of diarrhea in yaks.     

Dietary carbohydrate/lipid ratios and nutritional condition in juvenile southern rock lobster, Jasus edwardsii

The availability of formulated diets is considered economically imperative if rock lobster aquaculture, based on the collection and ongrowing of puerulus, is to progress. Central to the development of diets is an understanding of the nutritional requirements of the species. This study determined the effect of different dietary carbohydrate/lipid ratios (17:1, 5:1, 2:1, 0.8:1) on the growth and nutritional condition of juvenile southern rock lobsters, Jasus edwardsii, by measuring standard growth parameters, proximate composition of the whole body and digestive gland, and from a histological investigation of the digestive gland. Four replicate groups of eight lobsters (initial weight 5.08±0.98 g (mean±S.D.)) per diet treatment were held in 50 l tanks, in a recirculating system at 18 °C for 84 days. Maximum growth and the highest levels of lipid and dry matter in digestive glands and whole bodies was found in lobsters fed a diet containing 27% carbohydrate and 13.5% lipid (2:1 ratio) suggesting that of the four experimental diets, this diet provided the best balance of lipid and carbohydrate. Digestive gland histology supported this conclusion and lobsters fed low carbohydrate, high lipid diets were in the best nutritional condition, with high lipid accumulation, and structurally sound epithelial cells. Digestive gland epithelial cells of lobsters fed the high carbohydrate, low lipid diets were compressed, of inconsistent shape and size, with low lipid accumulation. Histology is therefore seen as a feasible method, in addition to growth and proximate composition data, to further examine the effect of diets in nutritional studies of crustaceans.     

Insect inhabitants of old larval galleries of<Emphasis Type="Italic"> Saperda populnea</Emphasis> (L.) (Coleoptera: Cerambycidae) in Bulgaria

The complex of insect inhabitants of old larval galleries of Saperda populnea (L.) (Coleoptera: Cerambycidae) was studied during the period 2000–2003 in 18 localities in Bulgaria. As a result, 32 insect species were reared from old S. populnea galls: Metopoplax origani (Kolenati) (Heteroptera: Lygaeidae), Malachius sp. (Coleoptera: Malachiidae), Agrilus pratensis pratensis (Ratzeburg) (Coleoptera: Buprestidae), Xylocleptes bispinus (Duftschmidt) (Coleoptera: Scolytidae), Crossocerus cetratus (Shuckard), Crossocerus megacephalus (Rossi), Crossocerus nigritus (Lepeletier & Brullé), Crossocerus acanthophorus (Kohl), Passaloecus brevilabris Wolf, Passaloecus gracilis (Curtis), Psenulus schencki (Tournier), Solierella compedita compedita (Piccioli), Trypoxylon figulus figulus (Linnaeus), Trypoxylon fronticorne fronticorne Gussakovskij (Hymenoptera: Crabronidae), Cleptes schmidti Linsenmaiser, Trichrysis cyanea (Linnaeus) (Hymenoptera: Chrysididae), Clistopyga incitator (Fabricius), Ctenochira sanguinatoria (Ratzeburg), Liotryphon crassisetus (Thomson), Nemeritis fallax (Gravenhorst), Mesochorus georgievi Schwenke, Campoletis sp. (Hymenoptera: Ichneumonidae), Charmon extensor (Linnaeus) (Hymenoptera: Braconidae), Eupelmus urozonus Dalman (Hymenoptera: Eupelmidae), Perilampus aeneus (Rossius) (Hymenoptera: Perilampidae), Trichiocampus grandis (Lepeletier) (Hymenoptera: Tenthredinidae), Janus luteipes (Lepeletier) (Hymenoptera: Cephidae), Cydia corollana (Hübner) (Lepidoptera: Tortricidae), Fiebrigella brevibucca (Duda) (Diptera: Chloropidae), Eustolomyia hilaris (Fallén) (Diptera: Anthomyiidae), Senotaina sp. (Diptera: Sarcophagidae) and Heringia vitripennis (Meigen) (Diptera: Syrphidae). Amongst them, 27 species were established as new inhabitants of old S. populnea galls. Five species (C. schmidti, N. fallax, M. georgievi, P. aeneus and J. luteipes) were recorded for the first time in Bulgaria. The dwellers of old S. populnea galls belong to the following ecological groups: insects using galls as a place to develop (3 species); insects nidificating in empty galls (10 species); insects using old galls as a pupating place (1 species); insects using empty galls as overwintering shelters (3 species); predators and parasites of primary dwellers of empty galls (13 species); or insects with uncertain ecological status (2 species).     

Ecology,physiology and genetics of a phycodnavirus infecting the noxious bloom-forming raphidophyte <Emphasis Type="Italic">Heterosigma akashiwo</Emphasis>

Abstract:   Heterosigma akashiwo virus (HaV) is a large icosahedral virus (∼0.2 μm) harboring a double-stranded DNA (dsDNA) genome (∼294 kbp). The virus is the only member of the genus Raphidovirus in the family Phycodnaviridae. Since its first discovery, a number of ecologic, physiologic and genetic studies about HaV have been conducted; especially, the relationship between H. akashiwo and HaV in nature was studied and viral infection is now regarded as a significant factor influencing the dynamics and termination of H. akashiwo blooms. HaV infection has considerable impacts on H. akashiwo populations in both aspects of fluctuation in biomass (quantity) and changes in clonal composition (quality). Partial sequencing of the HaV genome revealed that a number of genes showed considerable similarity to those of other protist-infecting viruses; still, the phylogenetic position of HaV suggested a number of enigmas in host–virus coevolution. Here are summarized the ecology, physiology and genetics of HaV especially from the viewpoint of the host–virus relationship.     

Detection of Colletotrichum coccodes from soil and potato tubers by conventional and quantitative real-time PCR

Colletotrichum coccodes is the causal agent of the potato blemish disease black dot. Two PCR primer sets were designed to sequences of the ribosomal internal transcribed spacer (ITS1 and ITS2) regions for use in a nested PCR. The genus-specific outer primers (Cc1F1/Cc2R1) were designed to regions common to Colletotrichum spp., and the species-specific nested primers (Cc1NF1/Cc2NR1) were designed to sequences unique to C . coccodes . The primer sets amplified single products of 447 bp (Cc1F1/Cc2R1) and 349 bp (Cc1NF1/Cc2NR1) with DNA extracted from 33 European and North American isolates of C. coccodes. The specificity of primers Cc1NF1/Cc2NR1 was confirmed by the absence of amplified product with DNA of other species representing the six phylogenetic groups of the genus Colletotrichum and 46 other eukaryotic and prokaryotic plant pathogenic species. A rapid procedure for the direct extraction of DNA from soil and potato tubers was used to verify the PCR assay for detecting C. coccodes in environmental samples. The limit of sensitivity of PCR for the specific detection of C. coccodes when inoculum was added to soils was 3·0 spores per g, or the equivalent of 0·06 microsclerotia per g soil, the lowest level of inoculum tested. Colletotrichum coccodes was also detected by PCR in naturally infested soil and from both potato peel and peel extract from infected and apparently healthy tubers. Specific primers and a TaqMan fluorogenic probe were designed to perform quantitative real-time (TaqMan) PCR to obtain the same levels of sensitivity for detection of C. coccodes in soil and tubers during a first-round PCR as with conventional nested PCR and gel electrophoresis. This rapid and quantitative PCR diagnostic assay allows an accurate estimation of tuber and soil contamination by C. coccodes .     

A rapid and miniaturized system using Alamar blue to assess fungal spore viability: implications for biosecurity

Long-lasting viable fungal spores are one of the important aspects in emergence, spread and disease development of pathogenic fungi. We developed a rapid and miniaturized system using Alamar Blue (resazurin dye; 7-hydroxy-3H-phenoxazin-3-one 10-oxide) for assessing fungal spore viability, using the ascomycete Leptosphaeria maculans (causing blackleg disease on canola) as a ‘model pathogen’. The assay is dependent on the metabolic activity of viable fungal spores to convert the dark blue of resazurin (maximum absorbance 605 nm) into the pink colour of resorufin (maximum absorbance 573 nm). The Alamar Blue assay uses an optimised micro-titre based format that was far superior for determining fungal spore viability in comparison with current conventional techniques including trypan blue staining, a TC10 cellometer cell counter, or by assessing germination of the spores under the microscope. This new assay was also more rapid and reproducible than current conventional tests to detect viable spores. Viable spores could be reliably detected within two hours. The successful application of the Alamar Blue assay to measure fungal spore viability in the current study has important benefits for biosecurity operations relating to faster and more reliable confirmation of viability of potential invasive exotic fungal pathogens and in minimising any consequent disease outbreaks. The effectiveness of the Alamar Blue assay was confirmed by successfully determining the relative retention times of viable L. maculans ascospores across a range of different potential spore-carrier materials, including steel, fabric, wood, paper, rubber and leather, over a time period of eight months. To further confirm the wide applicability of the Alamar Blue assay, it was successfully applied to detect viable spores of fungal pathogens of diverse taxonomic groups, including Kabatiella caulivora, Magnaporthe oryzae and Puccinia striiformis f.sp. tritici, and also of the yeast Saccharomyces cerevisiae.     

Impact of sulfur fertilization on distribution of sulfur fractions and use efficiency in blackgram in subtropical acidic soil of Assam,India

A field experiment was conducted to assess the effect of sulfur (S) fertilization on distribution of S in soil and use efficiency on blackgram in subtropical Inceptisol of acidic soil of Assam, India. Five levels of S were applied (0, 10, 20, 30 and 40 kg S ha^?1) along with recommended dose of nitrogen, phosphorus and potassium. Available S content gradually decreased with the advancement of crop growth stages and lowest value was observed at 60 DAS. Different S fractions were found to increase with increasing levels of S application and 40 kg S ha^?1 resulted the highest content for all S fractions. The grain and stover yield of blackgram increased significantly up to 20 kg S ha^?1 which was 95.69% higher over control. Agronomic efficiency, apparent S recovery and recovery efficiency of S were higher at 10 kg S ha^?1and found decreased with increase in level of S.