Impact of numerous larval diets on the biology of pink bollworm Pectinophora gossypiella(Saunders)(Lepidoptera:Gelechiidae)under laboratory conditions

Background Pink bollworm,Pectinophora gossypiella(Saunders)(Lepidoptera:Gelechiidae)has become a poten-tial pest of cotton by causing substantial yield losses around the world including Pakistan.Keeping in view the facts like limited research investigations,unavailability,and high cost of artificial diet’s constituents and their premixes,the present research investigations on the dietary aspect of P.gossypiella were conducted.The larvae of P.gossypiella were reared on different diets that were prepared using indigenous elements.The standard/laboratory diet com-prised of wheat germ meal 34.5 g,casein 30.0 g,agar–agar 20.0 g,sucrose 10.0 g,brewer’s yeast 5.0 g,α-cellulose 1.0 g,potassium-sorbate1.5 g,niplagin 0.5 g,decavitamin 0.01 g,choline-chloride 0.06 g,maize-oil 3.30 g,honey 2.0 g,and water 730.0 mL.Alternatives to cotton bolls and wheat germ meal were okra seed sprouts,okra fruit,cottonseed meal,and okra seed meals,which were included in the study to introduce an efficient and economic mass-rearing system.Results The larval development completed in 19.68d±0.05 d with a weight of 20.18mg±0.20 mg at the fourth instar fed on the cottonseed meal-based diet instead of wheat germ meal based diet.On the same diet,84.00%±4.00%,17.24 mg±0.03 mg,and 7.76d±0.06 d were recorded as pupae formation,pupal weight,and pupal duration,respectively.Adult emergence,76.00%±1.00%was recorded from pupae collected from larvae raised on cottonseed meal-based diet.These male and female moths lived for 40.25d±0.10 d,and 44.34d±0.11 d,respectively.Females deposited 21.28±0.04 eggs per day with the viability of 65.78%±0.14%.The larval mortal-ity at the fourth instar was 37.20%±1.36%and malformed pupation of 12.00%±1.41%was recorded.Replacement of wheat germ meal with that of local meals(cottonseed and okra seed)in the standard laboratory diet has saved 463.80 to 467.10 PKR with 1.62 to 1.63 cost economic returns,respectively.Conclusion This research is of novel nature as it provides a concise and workable system for the economic and suc-cessful rearing of P.gossypiella under laboratory conditions.

Antihyperglycemic effect of Hypericum perforatum ethyl acetate extract on streptozotocin-induced diabetic rats

Objective:To evaluate the antihyperglycemic activity of ethyl acetate extract of Hypericum perforatum(H.perforatum)in streptozotocin(STZ)-induced diabetic rats.Methods:Acute toxicity and oral glucose tolerance lest were performed in normal rats.Male albino rats were rendered diabetic by ST/(40 mg/kg,intraperitoneally).H.perforatum ethyl acetate extract was orally administered to diabetic rats at SO,100 and 200 mg/kg doses for 15 days to determine the antihyperglycemic activity.Biochemical parameters were determined at the end of the treatment.Results:H.perforatum ethyl acetate extract showed dose dependant fall in fasting blood glucose(FBG).After 30 min of extract administration,FBG was reduced significantly when compared with normal rats.H.perforatum ethyl acetate extract produced significant reduction in plasma glucose level,serum total cholesterol,triglycerides,glucose-6-phosphatase levels.Tissue glycogen content,HDL-cholesterol,glucose-6-phosphate dehydrogenase were significantly increased compared with diabetic control.No death or lethal effect was observed in the toxic study.Conclusions:The results demonstrate that H.perforatum ethyl acetate extract possesses potent antihyperglycemic activity in STZ induced diabetic rats.

Effect of Biophytum sensitivum on streptozotocin and nicotinamideinduced diabetic rats

Objective:To investigate the effect of aqueous solution of Biophytum sensUiuum leaf extract(BSEt)on normal and strcptozotocin(STZ)-nicotinamide-induced diabetic rats.Methods:Diabetes was induced in adult male Wislar rata by the administration of STZ-nicotinamide(40,110 mg/kg b.w.,respectively)intraperitoneally.BSEt(200 mg/kg)was administered to diabetic rats for 28 days.The effect of extract on blood glucose,plasma insulin,total haemoglobin,glycosylated haemoglobin,liver glycogen and carbohydrate melatxilism regulating enzymes of liver was studied in diabetic rats.Results:BSEt significantly reduced the blood glucose and glycosylated haemoglobin levels and significantly increased the total haemoglobin,plasma insulin and liver glycogen levels in diabetic rats.It also increased the hexokinase activity and decreased glucose-6-phosphatase,fructose-1,6-bisphosphatase activities in diabetic rats.Conclusions:The results of our study suggest that BSEt possesses a promising effecl on STZ-nicotinamide-induced diabetes.

Characterization and phylogenetic analysis of novel polyene type antimicrobial metabolite producing actinomycetes from marine sediments:Bay of Bengal India

Objective:To isolate and indentify the promising antimicrobial metabolite producing Streptomyces strains from marine sediment samples from Andraprudesh coast of India.Methods:Antagonistic aetinomycetes were isolated by starch casein agar medium and modified nutrient agar medium with 1%glucose used as a base for primary screening.Significant antimicrobial metabolite producing strains were selected and identified by using biochemical and 16S rDNA level.Minimum inhibitory concentrations of the organic extracts were done by using broth micro dilution method.Results:Among the 210 actinomyeetes,64.3%exhibited activity against Gram positive bacteria,48.5%showed activity towards Cram negative bacteria,38.8%exhibited both Cram positive and negative bacteria and 80.85%isolates revealed significant antifungal activity.However,five isolates AP-5,AP-18,AP-41 and AP-70 showed significant antimicrobial activity.The analysis of cell wall hydrolysates showed the presence of LL-diaminopimelic acid and glycine in all the isolates.Sequencing analysis indicated that the isolates shared 98.5%-99.8%sequence identity to the 16S rDNA gene sequences of the Streptomyces taxons.The antimicrobial substances were extracted using hexane and ethyl acetate from spent medium in which strains were cultivated at 30X for five days.The antimicrobial activity was assessed using broth micro dilution technique.Each of the culture extracts from these five strains showed a typical polyenelike property.The lowest minimum inhibitory concentrations of ethyl acetate extracts against Escherichia coli and Cumularia lunula were 67.5 and 125.0μg/mL,respectively.Conclusions:It can be concluded that hexane and ethyl acetate soluble extracellular products of novel isolates are effective against pathogenic bacteria and fungi.

Antibacterial activity of some actinomycetes from Tamil Nadu,India

Objective:To isolate novel actinomycetes and to evaluate their antibacterial activity.Methods:Three soil samples were collected from Vengodu(village)in Kanchipuram district,Tamil Nadu,India.Actinomycetes were isolated using serial dilution and plating method on actinomycetes isolation agar.Results:Totally 35 isolates were obtained on the basis of colony characteristics on actinomycetes isolation agar.All the isolates were screened for antibacterial activity by cross streak method.Medium and optimization of day were done for the potent strains using Nathan's agar well diffusion method.Isolation of bioactive compounds from significant active isolates was done by using different media.The most active isolate VAS 10 was identified as Actinobacterium Loyola PBT VAS 10(accession No.JF501398)using 16s rRNA sequence method.The hexane,ethyl acetate,dichloromethane and butanol extracts of VAS 10 were tested against bacteria.The maximum antibacterial activity was observed in dichloromethane and ethyl acetate;maximum zones of inhibition were observed against Enterococcus durans.The rRNA secondary structure and the restriction sites of Actinobacterium Loyola VAS 10 were predicted using Genebee and NEBCutter online tools respectively.Conclusions:The present study showed that among the isolated actinomycetes,Actinobacterium Loyola PBT VAS 10(accession No.JF501398)showed good antibacterial activity against the tested bacteria.

Cytotoxic (A549) and antimicrobial effects of Methylobacterium sp.isolate (ERI-135) from Nilgiris forest soil,India

Objective:To assess the antimicrobial and cytotoxic effects of Methylobacterium sp.isolated from soil sample of Doddabetta forest,Nilgiris,Western Ghats of Tamil Nadu.Methods:Isolation of Methylobacterium was performed from soils by serial dilution plate technique.The strain was grown in modified nutrient gulucose agar(MNGA)medium to study the morphology and biochemical characteristics.Methylobacterium sp.was screened for its antimicrobial activity against pathogenic bacteria and fungi.The strain was subjected to 16S rRNA analysis and was identified as Methylohacterium sp.The nucleotide sequence of the 16S rRNA gene of the isolate exhibited close similarity with other Methylobacterium sp.and has been submitted to Genbank.The antibacterial substances were extracted using chloroform and ethyl acetate from MNGA medium in which ERI-135 had grown for 5 d at 30℃.Cytotoxic effect was also studied.GC-MS analysis was carried out.The antimicrobial activity was assessed using broth micro dilution technique.Results:Ethyl acetate extract showed activity against bacteria such as Bacillus subtilis,Klebsiella pneumoniae(K.pneumoniae),Pseudomonas aeruginosa,Salmonella typkimurium,Shigella flexneri,Enterobacter aerogenes,Staphylococcus aureu and Staphylococcus epidermidis(S.epidermidis)and fungi such as,Candida albicans and Trichophyton rubrum.The lowest minimum inhibitory concentrations were:250μg/mL against 5.epidermidis and 250μg/mL against K.pneumonia.The isolate had the ability to produce enzymes such as protease.The exyract showed cytotoxic effect in human adenocarcinoma cancer cell line(A549).GC-MS analysis showed the presence of isovaleric acid(3.64%),2-Methylbulanoic acid(5.03%),isobutyramide(5.05%),N,Noimethylformamide-di-t-butylacetal(9.79%),benzeneacetamide(15.56%),octyl butyl phthalate(3.59%)and diisooctyl phthalate(5.79)in the extract.Conclusions:Methylobaeterium sp.(ERI-135)showed promising antibacterial and cytotoxic activity.This is the first report in the antimicrobial and cytotoxic effect of Methylobaeterium sp.

Larvicidal,ovicidal and repellent activities of marine sponge Cliona celata(Grant) extracts against Anopheles stephensi Liston(Diptera:Culicidae)

Obective:To evaluate the larvieidal.ovieidal and repellent properties of solvent extracts of marine sponge Cliona celala(C.celata)(Grant) against the malarial vector Anopheles stephensi(An.stephensi) Listen.Methods:Marine sponge C.celala was thoroughly washed with distilled water and shade dried for 48 h.Then the sponges were homogenized and extracted sequentially with hexane,ethyl acetate and methanol,Lanvicidal and ovicidal activities were tested at four different concentrations viz.,62.5.125.0.250.0 and 500.0 ppm.for repellent study extracts were taken in three different concentrations viz..5.0.2.5.1.0 mg/cm at.Results:Among the three solvent extracts of C.celata,methanol extract showed thc highest lanvieidal activity at 500 ppm against the fourth inslar larvae of An.stephensi.The LC_(50)and LC_(90) values of C.celata methanol extract were recorded as 80.61 and 220.81 ppm against An.stephensi larvae respectively.High ovieidal activity of 91.2%was recorded al 500 ppm concentration of methanol extract.The haxane extract was found to be the most effective protectant against the adult female mosquitoes of An.stephensi.The mean protection time recorded in hexane extract was up to 245 min at 5 mg/cm^2 dosage against An.Stephensi adults.Conclusions:The screening results suggest that the hexane and methanol extracts of C.celata arc promising in mosquito control.Considering these bioaclivitics.C.celata could the probed further to obtain some novel pesticidal molecules.

Ovicidal activity of Atalantia monophylla(L)Correa against Spodoptera litura Fab.(Lepidoptera:Noctuidae)

Objective:To evaluate the efficacy of Atalantia monophylla(A.monophylla)leaf in different solvent crude extracts and fractions against eggs of Spodoptera litura(S.litura).Methods:Hexane,ethyl acetate and chloroform solvent extracts of A.monophylla leaf and 12 fractions from hexane extract were screened at 5.0%t,2.5%,1.0%and 0.5%for crude extracts and 1000,500,250 and 125 mg/kg for fractions against the eggs of S.litura for the ovicidal activity.LC_(50)and LC_(90)were calculated using probit analysis.Results:Hexane crude extract showed maximum ovicidal activity of 61.94%at 5.0%concentration with a correlation value of r^2=0.81,and least LC_(50)value of3.06%.Hexane extract was fractionated using silica gel column chromatography and 12 fractions were obtained.Fraction 9 was active which showed maximum ovicidal activity of 75.61%at 1000mg/kg with the LC_(50)value of 318.65 mg/kg and LC_(90)value of 1473.31 mg/kg.In linear regression analysis,significant and high correlation(r^2=0.81%)was seen between concentration and ovicidal activity of hexane crude extracts and its active frsctien.Conclusions:As per our knowledge,this is the first report for ovicidal activity of A.monophylla against S.litura,A.monophylla could be used for the management of S.litura and other insect pests.

Effect of Ichnocarpus frutescens (L.) R.Br.hexane extract on preadipocytes viability and lipid accumulation in 3T3-L1 cells

Objective:To investigate the crude extracts of Ichnacarpus frutescens(I.frutescens) for antiobesity effect.Methods:Leaves of I.frutescens were sequentially extracted with hexane,ethyl acetate, and methanol and their effect on viability of 3T3-L1 preadipocytes were evaluated.Based on this the apoptosis on preadipocytes was confirmed by DMA fragmentation and LDH(Lactate dehydrogenase) leakage assays.Anti-adipogenesis was performed by oil red O(ORO) staining and free glycerol release in the medium of differentiated adipocytes.Results:The hexane extract of I.frutescens(IFHE) inhibited cell viability in a time- and dose-related manner.An increased release of LDH.as a marker of membrane integrity,was observed at a dose of 200μg/mL The discontinuous DMA fragments on agarose gel electrophoresis showed the apoptotic effect of the IFHE.Morphological observations of cells stained with ORG showed a decrease in cellular lipid content at the concentrations tested compared to the induced control cells.In the experiment of lipolytic activity,treatment with IFHE enhanced glycerol secretion with the rates of approximately 28%.55%,and 46%at the concentrations of 100,200 and 300μg/mL,respectively.Conclusions: The observed properties clearly revealed the medicinal property of I.frulescens in the treatment of obesity.

Larvicidal and knockdown effects of some essential oils against <i>Culex quinquefasciatus</i>Say, <i>Aedes aegypti</i>(L.) and <i>Anopheles stephensi</i>(Liston)

Efficacy of 25 essential oils was screened against filarial vector, Culex quinquefasciatus, for their larvicidal and knockdown effects in a preliminary study. Of these, 8 oils viz. calamus oil, cinnamon oil, citronella oil, clove oil, eucalyptus oil, lemon oil, mentha oil and orange oil exhibited 100% larvicidal activity at 1000 ppm and 100% knockdown effect at 10% concentration. These 8 oils were screened further against Cx. Quinquefasciatus, Aedes aegypti and Anopheles stephensi for their larvicial and knockdown effects at different concentrations. Mentha oil was the most promising against An. Stephensi and Ae. Aegypti recording LC50 and LC90 values of 39.74 and 115.67 ppm and 46.23 and 165.36 ppm, respectively for larvicidal activity. Calamus oil was the most effective against Cx. Quinquefasciatus with LC50 and LC90 values of 40.40, and 140.07 ppm, respectively for larvicidal activity. Orange oil showed the most potent knockdown effect with the KT50 and KT95 values of 27.44, 26.22 and 29.91 and 70.81, 65.33 and 68.57 min, against An. stephensi, Cx. Quinquefasciatus and Ae. Aegypti, respectively. The results clearly indicated that mentha oil and calamus oil were the most promising larvicides and orange oil had potent knockdown effect against the tested mosquito species. These oils could be used to develop a new formulation to control mosquitoes.

Effect of Selected Insecticides on Helicoverpa armigera Hubner (Lepidoptera: Noctuidae) on Tomato (Lycopersicon esculentum Miller) and Their Successful Management

Helicoverpa armigera H. is a polyphagous notorious pest of a number of economic crops including tomato. In tomato, it can reduce its yield as high as 70% due to fruit boring. Although some predators and parasitoids have been recorded in the field for their natural control, their manipulation to a successful crop management is still awaited. To overcome this pest, insecticides play a significant role in its effective crop management program globally. Tomato is an absolutely perishable commodity and needs the specified period required between the application, of conventional pesticides of organophosphate group, and consumption which can hardly be afforded. Therefore, nine insecticides from the new chemistries, which were relatively safer to human and environment, i.e. Coragen (chlorantraniliprole), steward (indoxacarb), Belt (flubendamide), Delegate (spintoram), Volium Flexy (chlorantraniliprole + thiamethaxim), Fipronil (grafter), Proclaim (emamectin benzoate), Pirate (chlofenapyr) and Lufenuron (lufenuron), were tested on tomato crop heavily infested by this pest in the farmers field in districts Lodhran and Bahawalpur (Pakistan) in the month of April 2014. The mortality was compared after 4, 7 and 10 days and treatments were compared with control as well with one another. The average maximum % mortality, i.e. 89.36 and 85.09 of the pest, was observed with Volium Flexy, and Delegate also worked well even after 7 and 10 days. Similarly, the results on the basis of damaged fruits and percent loss of yield pointed out that Chlorantraniliprole, Flubendiamide and Indoxacarb had resulted better as compared with others, although the difference was statistically non significant. These new chemistry pesticides are suggested to be used at tomato crop against Helicoverpa armigera which are best fitted in the IPM program for the control of the pest.

An unusual cause of late tracheostomy bleed

Human myiasis is common in the tropical and subtropical areas of the world. Tracheostomal myiasis is a rare entity, and only 7 such cases have been reported in literature. Our study emphasizes the importance of health education with regards to hygiene, sanitation and home tracheostomy tube care especially in immunocompromised individuals.

miRNA-24 and miRNA-466i-5p controls inflammation in rat hepatocytes

The discovery of microRNAs （miRNAs） is one of tbe most significant breakthroughs in recent decades, miRNAs are a class of small, endogenous, non-coding RNAs （19-24 nt） that negatively regulate the expression of target genes by binding to their 3＇ untranslated region and are involved in various biological processes. Three main approaches that are globally used for the detection of miRNA include direct cloning, forward genetics and computational approaches using bioinformatics.

Geographic variation in cypermethrin insecticide resistance and morphometry in Spodoptera litura(Fabricius)(Lepidoptera:Noctuidae)

Objective:To find out the most resistant strain of Spodoptera litura(S.litura)from Tamil Nadu,Kerala and Karnataka by synthetic insecticide treatment.Methods:Using leaf disc no-choice method,the insects were tested with different doses for pesticides.The LC_(50)and LC_(90)values were calculated by probit analysis.Results:In the insect bioassay,the cypermethrin insecticide showed significantly higher LC_(50)values of 14.699 g/L and 15.738 g/L against the Tamil Nadu and Kerala S.litura insect cultures respectively.The body length of 2nd,3rd and 4th instar larvae were significantly higher(P≤0.05)in TTP insect population[(19.2±2.3)mm,(28.05±3.20)mm,(36.1±2.0)mm],when compared with KTK[(18.5±2.7)mm,(23.38±2.00)mm,(31.75±2.70)mm]and control,KBB[(15.65±2.30)mm,(23.65±2.70)mm,(33.2±2.2)mm]populations.The body breadth of 3rd instar larvae was significantly higher(P≤0.05)in TTP insect population(4.9±1.1)mm,when compared with KTK(3.93±0.80)mm and control,KBB(3.65±0.70)mm populations.Conclusions:The present study clearly showed that field collected strains were highly resistant when compared to control.Based on our study,we conclude that reduced use of synthetic pesticides is necessary and IPM is a better way to reduce the development of pesticide resistance among strains of S.litura.

Phytopesticidal effects of Spilanthes acmella(L.)Murr.leaves on three economically important lepidopteran insect pests

Objective:To evaluate the antifeedant and larvicidal activities of dichloromethane(DCM),acetone,dimethylsulfoxide(DMSO)and aqueous extracts of Spilanthes acmella(S.acmella)leaves against Earias vitella,Helicoverpa armigera and Spodoptera litura.Methods:DCM,acetone,DMSO and aqueous extracts were studied using fruit and leaf discs no-choice method at 0.625%,1.25%,2.5%and 5%concentration.Results:All the extracts showed antifeedant and larvicidal activities against all the tested insects.However,maximum antifeedant activity was detected in DCM extract of S.acmella against all tested pest,followed by acetone,DMSO and aqueous extracts at 5%concentration.The leaves extract of S.acmella also showed larvicidal activity;maximum larval mortality was observed in DCM extract against Helicoverpa armigera(48.88%),Spodoptera litura(44.88%)and Earias vitella(75.11%)followed by acetone,DMSO and aqueous extracts at 5%concentration.Conclusions:DCM extract of S.acmella could be further purified to develop a formulation for eco-friendly pest control agent.