Search results for: pyrethroid insecticide

Authors: Mohd Shazad, Kamal Kumar Gupta

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Aedes aegypti (Diptera: Culicidae), commonly known as tiger mosquito is the vector of dengue fever, yellow fever, chikungunya and zika virus. In the absence of any effective vaccine against these diseases, control the mosquito population is the only promising mean to prevent the diseases. Currently used chemical insecticides cause environmental contamination, high mammalian toxicity and hazards to non-target organisms, insecticide resistance and vector resurgence. Present research work aimed to explore the potentials of phytochemicals present in the Ocimum sanctum in management of mosquito population. The leaves of Ocimum were extracted with ethanol by ‘cold extraction method’. 0-24h old fourth instar larvae of Aedes aegypti were treated with the extract of concentrations 50ppm, 100ppm, 200ppm and 400ppm for 24h. Survival, growth and development of the treated larvae were evaluated. The adults emerged from the treated larvae were used for the reproductive fitness studies. Our results indicate 77.2% mortality in the larvae exposed to 400 ppm. At lower doses, although there was no significant reduction in the survival after 24h however, it decreased during subsequent days of observations. In control experiments, no mortality was observed. It was also observed that the larvae survived after treatment showed severe growth and developmental abnormalities. There was significant increase in larval duration. In control, fourth instar moulted into pupa after 3 days while larvae treated with 400 ppm extract were moulted after 4.6 days. Larva-pupa intermediates and the pupa-adult intermediates were observed in many cases. The adults emerged from the treated larvae showed impaired mating and oviposition behaviour. The females exhibited longer preoviposition period, reduced oviposition rate and decreased egg output. GCMS analysis of the ethanol extract revealed presence of JH mimics and intermediates of JH biosynthetic pathway. Potentials of Ocimum sanctum in integrated vector management programme of Aedes aegypti were discussed.

Keywords: Aedes aegypti, Ocimum sanctum, oviposition, survival

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Authors: Swarupa Shashi Senivarapu Vemuri

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Curry leaf (Murraya koeinigii (L.)) exported from India had insecticide residues above maximum residue limits, which are hazardous to consumer health and caused rejection of the commodity at the point of entry in Europe and middle east resulting in a check on export of curry leaf. Hence to study current pesticide usage patterns in major curry leaf growing areas, a survey on pesticide use pattern was carried out in curry leaf growing areas in Guntur districts of Andhra Pradesh during 2014-15, by interviewing farmers growing curry leaf utilizing the questionnaire to assess their knowledge and practices on crop cultivation, general awareness on pesticide recommendations and use. Education levels of farmers are less, where 13.96 per cent were only high school educated, and 13.96% were illiterates. 18.60% farmers were found cultivating curry leaf crop in less than 1 acre of land, 32.56% in 2-5 acres, 20.93% in 5-10 acres and 27.91% of the farmers in more than 10 acres of land. Majority of the curry leaf farmers (93.03%) used pesticide mixtures rather than applying single pesticide at a time, basically to save time, labour, money and to combat two or more pests with single spray. About 53.48% of farmers applied pesticides at 2 days interval followed by 34.89% of the farmers at 4 days interval, and about 11.63% of the farmers sprayed at weekly intervals. Only 27.91% of farmers thought that the quantity of pesticides used at their farm is adequate, 90.69% of farmers had perception that pesticides are helpful in getting good returns. 83.72% of farmers felt that crop change is the only way to control sucking pests which damages whole crop. About 4.65% of the curry leaf farmers opined that integrated pest management practices are alternative to pesticides and only 11.63% of farmers felt natural control as an alternative to pesticides. About 65.12% of farmers had perception that high pesticide dose will give higher yields. However, in general, Curry leaf farmers preferred to contact pesticide dealers (100%) and were not interested in contacting either agricultural officer or a scientist. Farmers were aware of endosulfan ban 93.04%), in contrast, only 65.12, per cent of farmers knew about the ban of monocrotophos on vegetables. Very few farmers knew about pesticide residues and decontamination by washing. Extension educational interventions are necessary to produce fresh curry leaf free from pesticide residues.

Keywords: Curry leaf, decontamination, endosulfan, leaf roller, psyllids, tetranychid mite

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Authors: Girija Jha

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This paper investigates the need of cotton to integrate sustainability. The methodology used in the paper is to do secondary research to find out the various environmental implications of cotton as textile material across its life cycle and try to look at ways and possibilities of minimizing its ecological footprint. Cotton is called ‘The Fabric of Our Lives’. History is replete with examples where this fabric used to be more than a fabric of lives. It used to be a miracle fabric, a symbol India’s pride and social Movement of Swaraj, Gandhijee’s clarion call to self reliance. Cotton is grown in more than 90 countries across the globe on 2.5 percent of the world's arable land in countries like China, India, United States, etc. accounting for almost three fourth of global production. But cotton as a raw material has come under the scanner of sustainability experts because of myriad reasons a few have been discussed here. It may take more than 20,000 liters of water to produce 1kg of cotton. Cotton harvest is primarily done from irrigated land which leads to Salinization and depletion of local water reservoirs, e.g., Drying up of Aral Sea. Cotton is cultivated on 2.4% of total world’s crop land but accounts for 24% usage of insecticide and shares the blame of 11% usage of pesticides leading to health hazards and having an alarmingly dangerous impact on the ecosystem. One of the possible solutions to these problems as proposed was GM, Genetically Modified cotton crop. However, use of GM cotton is still debatable and has many ethical issues. The practice of mass production and increasing consumerism and especially fast fashion has been major culprits to disrupt this delicate balance. Disposable fashion or fast fashion is on the rise and cotton being one of the major choices adds on to the problem. Denims – made of cotton and have a strong fashion statement and the washes being an integral part of their creation they share a lot of blame. These are just a few problems listed. Today Sustainability is the need of the hour and it is inevitable to incorporate have major changes in the way we cultivate and process cotton to make it a sustainable choice. The answer lies in adopting minimalism and boycotting fast fashion, in using Khadi, in saying no to washed denims and using selvedge denims or using better methods of finishing the washed out fabric so that the environment does not bleed blue. Truly, the answer lies in integrating state of art technology with age old sustainable practices so that the synergy of the two may help us come out of the vicious circle.

Keywords: cotton, sustainability, denim, Khadi

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Authors: Godfrey Nsereko, Daniel Kadobera, Denis Okethwangu, Joyce Nguna, Alex Riolexus Ario

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Background: Malaria is a leading cause of morbidity and mortality in Uganda. In April 2018, malaria cases surged in Nwoya District, northern Uganda, exceeding the action thresholds. We investigated to assess the outbreak’s magnitude, identify transmission risk factors, and recommend evidence-based control measures. Methods: We defined a malaria case as onset of fever in a resident of Nwoya District with a positive Rapid Diagnostic Test or microscopy for malaria P. falciparum from 1 February to 22 May 2018. We reviewed medical records in all health facilities of affected sub-counties to find cases. In a case-control study, we compared exposure risk factors between 107 case-persons and 107 asymptomatic controls matched by age and village. We conducted entomological assessment on vector-density and behavior. Results: We identified 3,879 case-persons (attack rate [AR]=6.5%) and 2 deaths (case-fatality rate=5.2/10,000). Females (AR=8.1%) were more affected than males (AR=4.7%). Of all age groups, the 5-18 year age group (AR=8.4%) was most affected. Heavy rain started on 4 March; a propagated outbreak began during the week of 2 April. In the case-control study, 55% (59/107) of case-patients and 18% (19/107) of controls had stagnant water around households for several days following rainfall (ORM-H=5.6, 95%CI=3.0-11); 25% (27/107) of case-patients and 51% (55/107) of controls wore long-sleeve cloths during evening hours (ORM-H=0.30, 95%CI=0.20-0.60); 29% (31/107) of case-patients and 15% (16/107) of controls did not sleep under a long-lasting insecticide-treated net (LLIN) (ORM-H=2.3, 95%CI=1.1-4.9); 37% (40/107) of case-patients and 52% (56/107) of controls had ≥1 LLIN per 2 household members (ORM-H=0.54, 95%CI=0.30-0.97). Entomological assessment indicated active breeding sites; Anopheles gambiae sensu lato species were the predominant vector. Conclusion: Increased vector breeding sites after heavy rainfall, together with inadequate malaria preventive measures caused this outbreak. We recommended increasing coverage for LLINs and larviciding breeding sites.

Keywords: malaria outbreak, Plasmodium falciparum, global health security, Uganda

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Authors: Bunindro Nameirakpam, Sonia Sougrapakam, Shannon B. Olsson, Rajashekar Yallappa

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Natural sources for new pesticidal compounds hold promise in view of their eco-friendly nature, selectivity and mammalian safety. Despite a large number of plants that show insecticidal activity and diversity of natural chemistry with inherent eco-friendly nature, newer classes of insecticides have eluded discovery. Artemisia vulgaris, known as Mugwort, is a universal herb used for folk medicine and religious purposes throughout the ancient world. In India, the essential oils of Artemisia vulgaris are used for its insecticidal, anti parasiticidal and antimicrobial properties. Traditionally, the dried leaves of Artemisia vulgaris are used to repel insects as well as rats in and around the granaries in the North-East India. Artemisia vulgaris collected during November from different ecological sites were studied for the bio-pesticidal utility against the stored grain pests. The insecticidal activities were found in the crude extracts of n-hexane and methanol from the samples collected in Sikkim and Manipur respectively. Using silica gel column chromatography protocol, we have isolated one novel bioactive molecule from the aerial parts of Artemisia vulgaris L based on various physical-chemical and spectroscopic techniques (IR, 1H NMR, 13C NMR and mass). The novel bioactive molecule is highly toxic and very low concentration (4.35 µg/l) is needed to control the stored product insects. In additional experiment results clearly showed the involvement of sodium pumps inhibition in the insecticidal action of purified compound in the Sitophilus oryzae. The knockdown activity of the purified compound is concomitant with the in vivo inhibition of Na+/ K+- ATPase. Further, our study showed insignificant differences in the seed germination of control and the treated grains. The lack of adverse effect of the novel bioactive molecule on the seed germination is highly desirable for seed/grain protectant and showing the potential to be developed as possible natural fumigants for the control of stored grain pests. The novel bioactive molecule is selective insecticide with a high margin of safety to mammals and showed promise as novel biopesticide candidate for grain protection. It is believed that Bio-pesticides can serve as the most important pest management tools as far as global safety is concerned.

Keywords: Indian traditional plant, Artemisia vulgaris, bio-pesticides, Na+/ K+- ATPase, seed germination

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Authors: Vanitha Varadharaj, Vijalakshmi Krishnamurthy

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Annona squamosa Linn, commonly known as Sugar apple, belonging to the family Annonaceae, is said to show varied medicinal effects, including insecticide, antiovulatory and abortifacient. The alkaloid and flavonoids present in Annona squamosa leaf has proved to have antioxidant activity. The present work has been planned to investigate the effect of ethanolic leaf extract of Annona squamosa leaf on Den Induced wistar albino rats. The study was carried out to analyze the biochemical Parmeters like Total Proteins, Bilirubin, Enzymatic and Non –Enzymatic enzymes, Marker enzymes and Tumor markers in serum and also the histopathological studies in liver is carried out in control and DEN induced rats. Supplementation of ELAS (Ethanolic Leaf Extract Of Annona squamosa) reduced the liver weight and also reduced the tumour incidence. Chemoprevention group showed near normal values of bilirubin when compared with the control rats. Total protein was decreased in the cancer bearing group and on treatment with the extract the levels of protein were restored. Both in pre and post treatment group, the activities of enzymatic antioxidants such as superoxide dismutase, catalase, and Glutathione peroxidase were increased but in pre treated animals it was more effective than post treated animals. The non- enzymatic antioxidants such as vitamin C and vitamin E were brought back to normal level significantly in post and pre treated animals. Activities of marker enzymes such as SGOT, SGPT, ALP, γ GT were significantly elevated in the serum of cancer animals and the values returned to normal after treatment with the extract suggesting the hepato protective effect of the extract. Lipid peroxide was found to be elevated in the cancer induced group. This condition was brought back to the normal in the pre and post treated animals with ELAS. Histological examination also confirmed the anti- carcinogenic potential of ELAS, Cancer induced groups had a triple fold increase in their AFP values when compared to other groups. DEN treatment increased the level of AFP expression while ELAS partially counteracted the effect of it. So the scientific validation obtained from this study may pave way to many budding scientists to find new drugs from Annona squamosa for various ailments.

Keywords: annona squamosa, biochemical parmeters, cancer, leaf extract

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Authors: Leonardo Parra, Daniel Martínez, Jorge Pizarro, Fernando Ortega, Manuel Chacón-Fuentes, Andrés Quiroz

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Control of Dalaca pallens or blackworms, one of the most important hypogeous pest in grassland in southern Chile, is based on the use of broad-spectrum insecticides such as organophosphates and pyrethroids. However, the rapid development of insecticide resistance in field populations of this insect and public concern over the environmental impact of these insecticides has resulted in the search for other control methods. Specifically, the use of endophyte fungi for controlling pest has emerged as an interesting and promising strategy. Endophytes from ryegrass (Lolium perenne), establish a biotrophic relationship with the host, defined as mutualistic symbiosis. The plant-fungi association produces alkaloids where peramine is the main toxic substance against Listronotus bonariensis, the most important epigean pest of ryegrass. Nevertheless, the effect of peramina on others pest insects, such as D. pallens, to our knowledge has not been studied, and also its possible metabolization in the body of the larvae. Therefore, we addressed the following research question: Do larvae of D. pallens store peramine after consumption of ryegrass endophyte infected (E+)? For this, specimens of blackworms were fed with ryegrass plant of seven experimental lines and one commercial cultivar endophyte free (E-) sown at the Instituto de Investigaciones Agropecuarias Carillanca (Vilcún, Chile). Once the feeding period was over, ten larvae of each treatment were examined. Individuals were dissected, and their gut was removed to exclude any influence of remaining material. The rest of the larva's body was dried at 60°C by 24-48 h and ground into a fine powder using a mortar. 25 mg of dry powder was transferred to a microcentrifuge tube and extracted in 1 mL of a mixture of methanol:water:formic acid. Then, the samples were centrifuged at 16,000 rpm for 3 min, and the supernatant was colected and injected in the liquid chromatography of high resolution (HPLC). The results confirmed the presence of peramine in the larva's body of D. pallens. The insects that fed the experimental lines LQE-2 and LQE-6 were those where peramine was present in high proportion (0.205 and 0.199 ppm, respectively); while LQE-7 and LQE-3 obtained the lowest concentrations of the alkaloid (0.047 and 0.053 ppm, respectively). Peramine was not detected in the insects when the control cultivar Jumbo (E-) was tested. These results evidenced the storage and metabolism of peramine during consumption of the larvae. However, the effect of this alkaloid present in 'future ryegrass cultivars' (LQE-2 and LQE-6) on the performance and survival of blackworms must be studied and confirmed experimentally.

Keywords: blackworms, HPLC, alkaloid, pest

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Authors: Somayeh Khanjani, Samaneh Nabavi, Shirin Jalili

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Aluminum phosphide (ALP) is an inorganic phosphide used to control insects and is a highly effective insecticide and rodenticide used frequently to protect stored grain. Acute poisoning with this compound is common in some countries including India and Iran, and is a serious health problem. In Iran it was known as "rice tablet", for its use to preserve rice. Two kinds of rice tablets one being herbal while other containing 3g aluminum phosphide (AlP) are available for use in Iranian households to protect stored food grains from pests and rodents. The toxicity of Aluminum phosphide is attributed to the liberation of phosphine gas in contact with water or weak acid and is the major cause of poisoning and deaths. Rice tablet (Aluminum Phosphid) poisoning may be associated with serious and sometimes incurable complications. In 61.3% of patients were shown uniform ingestion. Vomiting was the most common symptoms reported by 96.4% patients. Agitation was reported in 36.9% and felling of thirsty in 27.9 %. Although many complications such as Hypotension, Adult Respiratory Distress Syndrome (ARDS), Acute Renal Failure (ARF) AND Multi Organ Failure (MOF) were the common complications observed in these patients, but the most lethal complication was Cardiac Arrhythmias occurred in 36.9% of cases. Abdominal pain in 31.4% of the patients, nausea in 79.4% of the patients and 41.1% of the patients showed metabolic acidosis. Suicidal intention was the most common cause of poisoning leading to deaths in 18.6% of the patients. Aluminum phosphide can cause either elevation, decrease or no change in electrolytes, bicarbonate and blood glucose level. The possible mechanism for changes in blood glucose levels are complex and depend on the balance of factors which increase its concentration and those which reduce it. AlP poisoning has been postulated to stimulate cortisol which leads to increasing blood level of cortisol, also it may cause stimulation of glucagon, and Adrenaline secretion; in addition, it can inhibit insulin synthesis which may lead to hyperglycemia. Another suggested mechanism of hyperglycemia is rennin activity in some cases, an increase in magnesium level of plasma and that of tissues, and high phosphate level. Although hyperglycemia is most frequent in this poisoning and also is known as a marker of poor prognostic, hypoglycemia in aluminum phosphide poisoning is a rare finding which may be so dangerous. Patients showed sever hypotension and sever acidosis in addition to sever hypoglycemia. The presenting features of AlP intoxication are rapid onset of shock, severe metabolic acidosis, cardiac dysrhythmias and adult respiratory distress syndrome (ARDS).

Keywords: aluminum phosphide (ALP), rice tablet, poisoning, phosphine gas

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Authors: Ante Turudic, Filip Varga, Zlatko Liber, Jernej Jakse, Zlatko Satovic, Ivan Radosavljevic, Martina Grdisa

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Microsatellites (SSRs) are highly informative repetitive sequences of 2-6 base pairs, which are the most used molecular markers in assessing the genetic diversity of plant species. Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip) is an outcrossing diploid (2n = 18) endemic to the eastern Adriatic coast and source of the natural insecticide pyrethrin. Due to the high repetitiveness and large size of the genome (haploid genome size of 9,58 pg), previous attempts to develop microsatellite markers using the standard methods were unsuccessful. A next-generation sequencing (NGS) approach was applied on genomic DNA extracted from fresh leaves of Dalmatian pyrethrum. The sequencing was conducted using NovaSeq6000 Illumina sequencer, after which almost 400 million high-quality paired-end reads were obtained, with a read length of 150 base pairs. Short reads were assembled by combining two approaches; (1) de-novo assembly and (2) joining of overlapped pair-end reads. In total, 6.909.675 contigs were obtained, with the contig average length of 249 base pairs. Of the resulting contigs, 31.380 contained one or multiple microsatellite sequences, in total 35.556 microsatellite loci were identified. Out of detected microsatellites, dinucleotide repeats were the most frequent, accounting for more than half of all microsatellites identifies (21,212; 59.7%), followed by trinucleotide repeats (9,204; 25.9%). Tetra-, penta- and hexanucleotides had similar frequency of 1,822 (5.1%), 1,472 (4.1%), and 1,846 (5.2%), respectively. Contigs containing microsatellites were further filtered by SSR pattern type, transposon occurrences, assembly characteristics, GC content, and the number of occurrences against the draft genome of T. cinerariifolium published previously. After the selection process, 50 microsatellite loci were used for primer design. Designed primers were tested on samples from five distinct populations, and 25 of them showed a high degree of polymorphism. The selected loci were then genotyped on 20 samples belonging to one population resulting in 17 microsatellite markers. Availability of codominant SSR markers will significantly improve the knowledge on population genetic diversity and structure as well as complex genetics and biochemistry of this species. Acknowledgment: This work has been fully supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir/ Sch. Bip.) insecticidal potential’ - (PyrDiv) (IP-06-2016-9034).

Keywords: genome assembly, NGS, SSR, Tanacetum cinerariifolium

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Authors: Maged El-Sayed Mohamed

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Eucalyptus camaldulensis is one on the most well-known species of the genus Eucalyptus in the Middle east, its importance relay on the high production of its unique volatile constituents which exhibits many medicinal and pharmacological activities. The gall-forming wasp (Leptocybe invasa) has recently come into sight as the main pest attacking E. camaldulensis and causing severe injury. The wasp lays its eggs in the petiole and midrib of leaves and stems of young shoots of E. camaldulensis, which leads to gall formation. Gall formation by L. invasa damages growing shoot and leaves of Eucalyptus, resulting in abscission of leaves and drying. AIM: This study is an attempt to investigate the effect of the gall wasp (Leptocybe invasa) attack on the volatile constitutes of E. camaldulensis. This could help in the control of this wasp through stimulating plant defenses or production of a new allelochemicals or insecticide. The study of volatile constitutes of Eucalyptus before and after attack by the wasp can help the re-use and recycle of the infected Eucalyptus trees for new pharmacological and medicinal activities. Methodology: The fresh gall wasp-attacked and healthy leaves (100 g each) were cut and immediately subjected to hydrodistillation using Clevenger-type apparatus for 3 hours. The volatile fractions isolated were analyzed using Gas chromatography/mass spectrometry (GC/MS). Kovat’s retention indices (RI) were calculated with respect to a set of co-injected standard hydrocarbons (C10-C28). Compounds were identified by comparing their spectral data and retention indices with Wiley Registry of Mass Spectral Data 10th edition (April 2013), NIST 11 Mass Spectral Library (NIST11/2011/EPA/NIH) and literature data. Results: Fifty-nine components representing 89.13 and 88.60% of the total volatile fraction content respectively were quantitatively analyzed. Twenty-six major compounds at an average concentration greater than 0.1 ± 0.02% have been used for the statistical comparison. From those major components, twenty-one were found in both the attacked and healthy Eucalyptus leaves’ fractions in different concentration and five components, mono terpene p-Mentha-2-4(8) diene and the sesquiterpenes δ-elemene, β-elemene, E-caryophyllene and Bicyclogermacrene, were unique and only produced in the attacked-leaves’ fraction. CONCLUSION: Newly produced components or those commonly found in the volatile fraction and changed in concentration could represent a part of the plant defense mechanisms or might be an element of the plant allelopathic and communication mechanisms. Identification of the components of the gall wasp-damaged leaves can help in their recycling for different physiological, pharmacological and medicinal uses.

Keywords: Eucalyptus camaldulensis, eucalyptus recycling, gall wasp, Leptocybe invasa, plant defense mechanisms, Terpene fraction

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Authors: Fouzia Khanam, Tridib Chowdhury, Belal Hossain, Sajedur Rahman, Mahfuzar Rahman

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Introduction: Over the last decades, the world has achieved a noticeable success in preventing malaria. Nevertheless, malaria, a vector-borne infectious disease, remains a major public health burden globally as well as in Bangladesh. To achieve the goal of eliminating malaria, BRAC, a leading organization of Bangladesh in collaboration with government, is distributing free LLIN to the 13 endemic districts of the country. The study was conducted with the aim of assessing the gap between coverage, access, and utilization of LLIN among the people of the 13 malaria endemic districts of Bangladesh. Methods: This baseline study employed a community cross-sectional design triangulated with qualitative methods to measure households’ ownership, access and use of 13 endemic districts. A multistage cluster random sampling was employed for the quantitative part and for qualitative part a purposive sampling strategy was done. Thus present analysis included 2640 households encompassing a total of 14475 populations. Data were collected using a pre-tested structured questionnaire through one on one face-to-face interview with respondents. All analyses were performed using STATA (Version 13.0). For the qualitative part participant observation, in-depth interview, focus group discussion, key informant interview and informal interview was done to gather the contextual data. Findings: According to our study, 99.8% of households possessed at least one-bed net in both study areas. 77.4% households possessed at least two LLIN and 43.2% households had access to LLIN for all the members. So the gap between coverage and access is 34%. 91.8% people in the 13 districts and 95.1% in Chittagong Hill Tracts areas reported having had slept under a bed net the night before interviewed. And despite the relatively low access, in 77.8% of households, all the members were used the LLIN the previous night. This higher utilization compared to access might be due to the increased awareness among the community people regarding LLIN uses. However, among those people with sufficient access to LLIN, 6% of them still did not use the LLIN which reflects the behavioral failure that needs to be addressed. The major reasons for not using LLIN, identified by both qualitative and quantitative findings, were insufficient access, sleeping or living outside the home, migration, perceived low efficacy of LLIN, fear of physical side effects or feeling uncomfortable. Conclusion: Given that LLIN access and use was a bit short of the targets, it conveys important messages to the malaria control program. Targeting specific population segments and groups for achieving expected LLIN coverage is very crucial. And also, addressing behavior failure by well-designed behavioral change interventions is mandatory.

Keywords: long lasting insecticide net, malaria, malaria control programme, World Health Organisation

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Authors: Abhishek Sraw, Amit Sobti, Yamini Pandey, R. K. Wanchoo, Amrit Pal Toor

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Monocrotophos (MCP) is a widely used pesticide in India, which belong to an extremely toxic organophosphorus family, is persistent in nature and its toxicity is widely reported in all environmental segments in the country. Advanced Oxidation Process (AOP) is a promising solution to the problem of water pollution. TiO₂ is being widely used as a photocatalyst because of its many advantages, but it has a large band gap, due to which it is modified using metal and nonmetal dopant to make it active under sunlight and visible light. The use of nanosized powdered catalysts makes the recovery process extremely complicated. Hence the aim is to use low cost, easily available, eco-friendly clay material in form of bead as the support for the immobilization of catalyst, to solve the problem of post-separation of suspended catalyst from treated water. A recirculation type photocatalytic reactor (RTPR), using ultraviolet light emitting source (blue black lamp) was designed which work effectively for both suspended catalysts and catalyst coated clay beads. The bare, TiO₂ and W-TiO₂ coated clay beads were characterized by scanning electron microscopy (SEM), electron dispersive spectroscopy (EDS) and N₂ adsorption–desorption measurements techniques (BET) for their structural, textural and electronic properties. The study involved variation of different parameters like light conditions, recirculation rate, light intensity and initial MCP concentration under UV and sunlight for the degradation of MCP. The degradation and mineralization studies of the insecticide solution were performed using UV-Visible spectrophotometer, and COD vario-photometer and GC-MS analysis respectively. The main focus of the work lies in checking the recyclability of the immobilized TiO₂ over clay beads in the developed RTPR up to 30 continuous cycles without reactivation of catalyst. The results demonstrated the economic feasibility of the utilization of developed RTPR for the efficient purification of pesticide polluted water. The prepared TiO₂ clay beads delivered 75.78% degradation of MCP under UV light with negligible catalyst loss. Application of W-TiO₂ coated clay beads filled RTPR for the degradation of MCP under sunlight, however, shows 32% higher degradation of MCP than the same system based on undoped TiO₂. The COD measurements of TiO₂ coated beads led to 73.75% COD reduction while W-TiO₂ resulted in 87.89% COD reduction. The GC-MS analysis confirms the efficient breakdown of complex MCP molecules into simpler hydrocarbons. This supports the promising application of clay beads as a support for the photocatalyst and proves its eco-friendly nature, excellent recyclability, catalyst holding capacity, and economic viability.

Keywords: immobilized clay beads, monocrotophos, recirculation type photocatalytic reactor, TiO₂

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Authors: Sunil Kumar Ghosh

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Tomato (Lycopersicon esculentus L.) is an annual vegetable crop grown in the sub-Himalayan region of north east India throughout the year except rainy season in normal field cultivation. The crop is susceptible to various insect pests of which whitefly (Bemesia tabaci Genn.) causes heavy damage. Thus, a study on its occurrence and sustainable management is needed for successful cultivation. The pest was active throughout the growing period. During 38th standard week to 41st standard week that is during 3rd week of September to 2nd week of October minimum population was observed. The maximum population level was maintained during 11th standard week to 18th standard week that is during 2nd week of March to 3rd week of March with peak population (0.47/leaf) was recorded. Weekly population counts on white fly showed non-significant negative correlation (p=0.05) with temperature and weekly total rainfall where as significant negative correlation with relative humidity. Eight treatments were taken to study the management of the white fly pest such as botanical insecticide azadirachtin botanical extracts, Spilanthes paniculata flower, Polygonum hydropiper L. flower, tobacco leaf and garlic and mixed formulation like neem and floral extract of Spilanthes were evaluated and compared with the ability of acetamiprid. The insectide acetamiprid was found most lethal against whitefly providing 76.59% suppression, closely followed by extracts of neem + Spilanthes providing 62.39% suppression. Spectophotometric scanning of crude methanolic extract of Polygonum flower showed strong absorbance wave length between 645-675 nm. Considering the level of peaks of wave length the flower extract contain some important chemicals like Spirilloxanthin, Quercentin diglycoside, Quercentin 3-O-rutinoside, Procyanidin B1 and Isorhamnetin 3-O-rutinoside. These chemicals are responsible for pest control. Spectophotometric scanning of crude methanolic extract of Spilanthes flower showed strong absorbance wave length between 645-675 nm. Considering the level of peaks of wave length the flower extract contain some important chemicals of which polysulphide compounds are important and responsible of pest control. Neem and Spilanthes individually did not produce good results but when used as a mixture they recorded better results. Highest yield (30.15 t/ha) were recorded from acetamiprid treated plots followed by neem + Spilanthes (27.55 t/ha). Azadirachtin and Plant extracts are biopesticides having less or no hazardous effects on human health and environment. Thus they can be incorporated in IPM programmes and organic farming in vegetable cultivation.

Keywords: biopesticides, organic farming, seasonal fluctuation, vegetable IPM

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Authors: Shiva Mardani, Mehdi Nasr-Esfahani, Majid Olia, Hamid Molahosseini, Hamed Hassanzadeh Khankahdani

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Plant-parasitic nematodes cause serious diseases on plants and effectively reduce food production in quality and quantity worldwide, with at least 17 nematode species in the three important and major genera, including Meloidogyne, Heterodera, and Pratylenchus. Root-knot nematodes (RKN), Meloidogyne spp. with the dominant species, Meloidogynejavanica, are considered as the important plant pathogens of agricultural products globally. The hosts range can be vegetables, bedding plants, grasses, shrubs, numerous weeds, and trees, including forests. In this study, chemical management was carried out on RKN, M. javanica, to investigate the efficacy of Iranian Abamectin insecticide product [acaricide Abamectin (Vermectin® 2% EC, Gyah Corp., Iran)] verses imported normal Abamectin available in the Iran markets [acaricide Abamectin (Vermectin® 1.8% EC, Cropstar Chemical Industry Co., Ltd.)] each of which at the rate of 8 L./ha, on Tomatoes, Solanumlycopersicum L., (No. 29-41, Dutch company Siemens) as a test plant, and the controls (infested to RKN and without any chemical pesticides treatments); and (sterile soil without any RKN and chemical pesticides treatments) at the greenhouse in Isfahan, Iran. The trails were repeated thrice. The results indicated a highly significant reduction in RKN population and an increase in biomass parameters at 1% level of significance, respectively. Relatively similar results were obtained in all the three experiments conducted on tomato root-knot nematodes. The treatments of Gyah-Abamectin (51.6%) and external Abamectin (40.4%) had the highest to least effect on reducing the number of larvae in the soil compared to the infected controls, respectively. Gyah-Abamectin by 44.1% and then external one by 31.9% had the highest effect on reducing the number of larvae and eggs in the root and 31.4% and 24.1% reduction in the number of galls compared to the infected controls, respectively. Based on priority, Gyah-Abamectin (47.4 % ) and external Abamectin (31.1 %) treatments had the highest effect on reducing the number of egg- masses in the root compared to the infected controls, with no significant difference between Gyah-Abamectin and external Abamectin. The highest reproduction of larvae and egg in the root was observed in the infected controls (75.5%) and the lowest in the healthy controls (0.0%). The highest reduction in the larval and egg reproduction in the roots compared to the infected controls was observed in Gyah-Abamectin and the lowest in the external one. Based on preference, Gyah-Abamectin (37.6%) and external Abamectin (26.9%) had the highest effect on the reduction of the larvae and egg reproduction in the root compared to the infected controls, respectively. Regarding growth parameters factors, the lowest stem length was observed in external Abamectin (51.9 cm), with nosignificantly different from Gyah-Abamectin and healthy controls. The highest root fresh weight was recorded in the infected controls (19.81 gr.) and the lowest in the healthy ones (9.81 gr.); the highest root length in the healthy controls (22.4 cm), and the lowest in the infected controls and external Abamectin (12.6 and 11.9 cm), respectively. Conclusively, the results of these three tests on tomato plants revealed that Gyah-Abamectin 2% compared to external Abamectin 1.8% is competitive in the chemical management of the root nematodes of these types of products and is a suitable alternative in this regard.

Keywords: solanum lycopersicum, vermectin, biomass, tomato

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Authors: Filip Varga, Nina Jeran, Martina Biosic, Zlatko Satovic, Martina Grdisa

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Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.), a species endemic to the eastern Adriatic coastline, is the source of natural insecticide pyrethrin. Pyrethrin is a mixture of six compounds (pyrethrin I and II, cinerin I and II, jasmolin I and II) that exhibits high insecticidal activity with no detrimental effects to the environment. A recently optimized matrix-solid phase dispersion method (MSPD), using florisil as the sorbent, acetone-ethyl acetate (1:1, v/v) as the elution solvent, and sodium sulfate anhydrous as the drying agent was utilized to extract the pyrethrins from 10 wild populations (20 individuals per population) distributed along the Croatian coast. All six components in the extracts were qualitatively and quantitatively determined by high-performance liquid chromatography with a diode array detector (HPLC-DAD). Pearson’s correlation index was calculated between pyrethrin compounds, and differences between the populations using the analysis of variance were tested. Additionally, the correlation of each pyrethrin component with spatio-ecological variables (bioclimate, soil properties, elevation, solar radiation, and distance from the coastline) was calculated. Total pyrethrin content ranged from 0.10% to 1.35% of dry flower weight, averaging 0.58% across all individuals. Analysis of variance revealed significant differences between populations based on all six pyrethrin compounds and total pyrethrin content. On average, the lowest total pyrethrin content was found in the population from Pelješac peninsula (0.22% of dry flower weight) in which total pyrethrin content lower than 0.18% was detected in 55% of the individuals. The highest average total pyrethrin content was observed in the population from island Zlarin (0.87% of dry flower weight), in which total pyrethrin content higher than 1.00% was recorded in only 30% of the individuals. Pyrethrin I/pyrethrin II ratio as a measure of extract quality ranged from 0.21 (population from the island Čiovo) to 5.88 (population from island Mali Lošinj) with an average of 1.77 across all individuals. By far, the lowest quality of extracts was found in the population from Mt. Biokovo (pyrethrin I/II ratio lower than 0.72 in 40% of individuals) due to the high pyrethrin II content typical for this population. Pearson’s correlation index revealed a highly significant positive correlation between pyrethrin I content and total pyrethrin content and a strong negative correlation between pyrethrin I and pyrethrin II. The results of this research clearly indicate high intra- and interpopulation diversity of Dalmatian pyrethrum with regards to pyrethrin content and composition. The information obtained has potential use in plant genetic resources conservation and biodiversity monitoring. Possibly the largest potential lies in designing breeding programs aimed at increasing pyrethrin content in commercial breeding lines and reintroduction in agriculture in Croatia. Acknowledgment: This work has been fully supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir/ Sch. Bip.) insecticidal potential’ - (PyrDiv) (IP-06-2016-9034).

Keywords: Dalmatian pyrethrum, HPLC, MSPD, pyrethrin

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Authors: M. D. Nirmani, K. L. N. Perera, G. H. Galhena

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Dengue has become one of the most important arbo-viral disease in all tropical and subtropical regions of the world. Aedes aegypti, is the principal vector of the virus, vary in both epidemiological and behavioral characteristics, which could be finely measured through DNA sequence comparison at their population level. Such knowledge in the population differences can assist in implementation of effective vector control strategies allowing to make estimates of the gene flow and adaptive genomic changes, which are important predictors of the spread of Wolbachia infection or insecticide resistance. As such, this study was undertaken to investigate the phylogenetic relationships of Ae. aegypti from Galle and Colombo, Sri Lanka, based on the ribosomal protein region which spans between two exons, in order to understand the geographical distribution of genetically distinct mosquito clades and its impact on mosquito control measures. A 320bp DNA region spanning from 681-930 bp, corresponding to the ribosomal protein, was sequenced in 62 Ae. aegypti larvae collected from Galle (N=30) and Colombo (N=32), Sri Lanka. The sequences were aligned using ClustalW and the haplotypes were determined with DnaSP 5.10. Phylogenetic relationships among haplotypes were constructed using the maximum likelihood method under Tamura 3 parameter model in MEGA 7.0.14 including three previously reported sequences of Australian (N=2) and Brazilian (N=1) Ae. aegypti. The bootstrap support was calculated using 1000 replicates and the tree was rooted using Aedes notoscriptus (GenBank accession No. KJ194101). Among all sequences, nineteen different haplotypes were found among which five haplotypes were shared between 80% of mosquitoes in the two populations. Seven haplotypes were unique to each of the population. Phylogenetic tree revealed two basal clades and a single derived clade. All observed haplotypes of the two Ae. aegypti populations were distributed in all the three clades, indicating a lack of genetic differentiation between populations. The Brazilian Ae. aegypti haplotype and one of the Australian haplotypes were grouped together with the Sri Lankan basal haplotype in the same basal clade, whereas the other Australian haplotype was found in the derived clade. Phylogram showed that Galle and Colombo Ae. aegypti populations are highly related to each other despite the large geographic distance (129 Km) indicating a substantial genetic similarity between them. This may have probably arisen from passive migration assisted by human travelling and trade through both land and water as the two areas are bordered by the sea. In addition, studied Sri Lankan mosquito populations were closely related to Australian and Brazilian samples. Probably this might have caused by shipping industry between the three countries as all of them are fully or partially enclosed by sea. For example, illegal fishing boats migrating to Australia by sea is perhaps a good mean of transportation of all life stages of mosquitoes from Sri Lanka. These findings indicate that extensive mosquito migrations occur between populations not only within the country, but also among other countries in the world which might be a main barrier to the successful vector control measures.

Keywords: Aedes aegypti, dengue control, extensive mosquito migration, haplotypes, phylogeny, ribosomal protein

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Authors: Nautiyal Ruchira, Nautiyal Hemant, Chaudhury Devnanda, Bhargava Surbhi, Chauhan Nidhi

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Introduction: South East Asian region contributes 2.5 million cases of malaria each year to the global burden of 300 to 500 million of which 76% is reported from India. Government of India launched a national program almost half a century ago, still malaria remains a major public health challenge. Pregnant women are more susceptible to severe malaria and its fetomaternal complications. Inadequate surveillance and under-reporting underestimates the problem. Aim: Present study aimed to analyze the clinical course and pattern of malaria during pregnancy and to study the feto-maternal outcome. Methodology: This is a prospective observational study carried out at Himalayan Institute of Medical Sciences – a tertiary care center in the sub-Himalayan state of Uttarakhand, Northern India. All the pregnant women with malaria and its complications were recruited in the study during 2009 to 2014 which included referred cases from the state of western Uttar Pradesh. A thorough history and clinical examination were carried out to assess maternal and fetal condition. Relevant investigations including haemogram, platelet count, LFT, RFT, and USG was done. Blood slides and rapid diagnostic tests were done to diagnose the type of malaria.The primary outcomes measured were the type of malaria infection, maternal complications associated with malaria, outcome of pregnancy and effect on the fetus. Results: 67 antenatal cases with malaria infection were studied. 71% patients were diagnosed with plasmodium vivax infection, 25% cases were plasmodium falciparum positive and in 3% cases mixed infection was found. 38(56%) patients were primigravida and 29(43%) were multiparous. Most of the patients had already received some treatment from their local doctors and presented with severe malaria with the complications. Thrombocytopenia was the commonest manifestation seen in 35(52%) patients, jaundice in 28%, severe anemia in 18%, and severe oligohydramnios in 10% and renal failure in 6% cases. Regarding pregnancy outcome there were 44 % preterm deliveries, 22% had IUFD and abortions in 6% cases.20% of newborn were low birth weight and 6% were IUGR. There was only one maternal death which occurred due to ARDS in falciparum malaria. Although Plasmodium vivax was the main parasite considering the severity of clinical presentation, all the patients received intensive care. As most of the patients had received chloroquine therapy hence they were treated with IV artesunate followed by oral artemesinin combination therapy. Other therapies in the form of packed RBC’s and platelet transfusions, dialysis and ventilator support were provided when required. Conclusion: Even in areas with annual parasite index (API) less than 2 like ours, malaria in pregnancy could be an alarming problem. Vivax malaria cannot be considered benign in pregnancy because of high incidence of morbidity. Prompt diagnosis and aggressive treatment can reduce morbidity and mortality significantly. Increased community level research, integrating ANC checkups with the distribution of insecticide-treated nets in areas of high endemicity, imparting education and awareness will strengthen the existing control strategies.

Keywords: severe malaria, pregnancy, plasmodium vivax, plasmodium falciparum

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Authors: Vijay Kumar, G. K. Grewal, Prasad S. Burange

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India is one of the largest cultivators of cotton in the world. Among the various constraints, insect pests are posing a major hurdle to the success of cotton cultivation. Various bollworms, including the pink bollworm, Pectinophora gossypiella (Saunders), cause serious losses in India, China, Pakistan, Egypt, Brazil, tropical America, and Africa, etc. Bt cotton cultivars having Cry genes were introduced in India in 2002 (Cry1Ac) and 2006 (Cry1Ac+ Cry2Ab) for control of American, spotted, and pink bollworms. Pink bollworm (PBW) larvae infest flowers, squares, and bolls. Larva burrows into flowers and bolls to feed on pollen and seeds, respectively. It has a shorter lifecycle and more generations per year, so it develops resistance more quickly than other bollworms. Further, it has cryptic feeding sites, i.e., flowers and bolls/seeds, so it is not exposed to harsh environmental fluctuations and insecticidal applications. The cry toxin concentration is low in its feeding sites, i.e., seeds and flowers of cotton. The use of insecticide and Bt cotton is the primary control measure that has been successful in limiting the damage of PBW. But with the passage of time, it has developed resistance against insecticides and Bt cotton. However, the use of insecticides increases chemical control costs while causing secondary pest problems and environmental pollution. Extensive research has indicated that monitoring and control measures such as biological, cultural, chemical, and host plant resistance methods can be integrated for effective PBW management. The potential of various biological control organisms needs to be explored. The impact of transgenic cotton on non-target organisms, particularly natural enemies, which play an important role in pest control, is still being debated. According to some authors, Bt crops have a negative impact on natural enemies, particularly parasitoids. An experiment was carried out in the Integrated Pest Management Laboratory of the Department of Entomology, Punjab Agricultural University, Ludhiana, Punjab, India, to study the natural parasitization of PBW on Bt cotton in 2022. A large population of larvae of PBW were kept individually in plastic containers and fed with cotton bolls until the emergence of a parasitoid cocoon. The first cocoon of the parasitoid was observed on October 25, 2022. Symptoms of parasitization were never seen on larvae. Larvae stopped feeding and became inactive before the emergence of parasitoids for pupation. Grub makes its way out of larvae by making a hole in the integument, and immediately after coming out, it spins the cocoon. The adult parasitoid emerged from the cocoon after eight days. The parasitoids that emerged from the cocoon were identified as Cotesia (Braconidae: Hymenoptera) based on the features of the adult. Out of 475 larvae of PBW, 87 were parasitized, with 18.31% of parasitization. Out of these, 6.73% were first instar, 10.52% were second instar, and 1.05% were third instar larvae of PBW. No parasitization was observed in fourth instar larvae. Parasitoids were observed during the fag end of cropping season and mostly on the earlier instars. It is concluded that the potential of Cotesia may be explored as a biological control agent against PBW, which is safer to human beings, environment and non-taraltoget organisms.

Keywords: biocontrol, Bt cotton, Cotesia, Pectinophora gossypiella

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Authors: Siddharudha B. Patil

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Cotton is a premier commercial fibre crop of India subjected to ravages of insect pests. Sucking pests viz thrips, Thrips tabaci,(lind) leaf hopper Amrsca devastance,(dist) miridbug, Poppiocapsidea beseratense (Dist) and bollworms continue to inflict damage Bt Cotton right from seeding stage. Their infestation impact cotton yield to an extent of 30-40 percent. Chemical control is still adoptable as one of the techniques for combating these pests. Presently, growers have many challenges in selecting effective chemicals which fit in with an integrated pest management. Spinetoram has broad spectrum with excellent insecticidal activity against both sucking pests and bollworms. Hence, it is expected to make a great contribution to stable production and quality improvement of agricultural products. Spinetoram is a derivative of biologically active substances (Spinosyns) produced by soil actinomycetes, Saccharopolypara spinosa which is semi synthetic active ingredient representing Spinosyn chemical class of insecticide and has demonstrated higher level of efficacy with reduced risk on beneficial arthropods. The efforts were made in the present study to test the efficacy of Spinetoram against sucking pests and bollworms in comparison with other insecticides in Bt Cotton under field condition. Field experiment was laid out during 2013-14 and 2014-15 at Agricultural Research station Dharwad (Karnataka-India) in a randomized block design comprising eight treatments and three replications. Bt cotton genotype, Bunny BG-II was sown in a plot size of 5.4 m x5.4 m. Recommend agronomical practices were followed. The Spinetoram 12% SC alone and incombination with sulfaxaflore with varied dosages against pest complex was tested. Performance was compared with Spinosad 45% SC and thiamethoxam 25% WG. The results of consecutive seasons revealed that nonsignificant difference in thrips and leafhopper population and varied significantly after 3 days of imposition. Among the treatments, combiproduct, Spinetoram 10%WG + Sulfoxaflor 30% WG@ 140 gai/ha registered lowest population of thrips (3.91/3 leaves) and leaf hoppers (1.08/3 leaves) followed by its lower dosages viz 120 gai/ha (4.86/3 leaves and 1.14/3 leaves of thrips and leaf hoppers, respectively) and 100 gai/ha (6.02 and 1.23./3 leaves of thrips and leaf hoppers respectively) being at par, significantly superior to rest of the treatments. On the contrary, the population of thrips, leaf hopper and miridbugs in untreated control was on higher side. Similarly the higher dosage of Spinetoram 10% WG+ Sulfoxaflor 30% WG (140 gai/ha) proved its bioefficacy by registering lowest miridbug incidence of 1.70/25 squares, followed by its lower dosage (1.78 and 1.83/25 squares respectively) Further observation made on bollworms incidence revealed that the higher dosage of Spinetoram 10% WG+Sulfoxaflor 30% WG (140 gai/ha) registered lowest percentage of boll damage (7.22%), more number of good opened bolls (36.89/plant) and higher seed cotton yield (19.45q/ha) followed by rest of its lower dosages, Spinetoram 12% SC alone and Spinosad 45% SC being at par significantly superior to rest of the treatments. However, significantly higher boll damage (15.13%) and lower seed cotton yield (14.45 q/ha) was registered in untreated control. Thus Spinetoram10% WG+Sulfoxaflor 30% WG can be a promising option for pest management in Bt Cotton.

Keywords: Spinetoram10% WG+Sulfoxaflor 30% WG, sucking pests, bollworms, Bt cotton, management

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Authors: Ajay Kumar Pandey, Mayank Kumar

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White grub (Coleoptera: Scarabaeidae) is a major destructive pest in western Himalayan region of Uttarakhand. Beetles feed on apple, apricot, plum, walnut etc. during night while, second and third instar grubs feed on live roots of cultivated as well as non-cultivated crops. Collection and identification of scarab beetles through light trap was carried out at Crop Research Centre, Govind Ballab Pant University Pantnagar, Udham Singh Nagar (Uttarakhand) during 2018. Field trials were also conducted in 2018 to evaluate pre and post sown application of different insecticides against the white grub infesting soybean. The insecticides like Carbofuran 3 Granule (G) (750 g a.i./ha), Clothianidin 50 Water Dispersal Granule (WG) (120 g a.i./ha), Fipronil 0.3 G (50 g a.i./ha), Thiamethoxam 25 WG (80 g a.i./ha), Imidacloprid 70 WG (300 g a.i./ha), Chlorantraniliprole 0.4% G(100 g a.i./ha) and mixture of Fipronil 40% and Imidacloprid 40% WG (300 g a.i./ha) were applied at the time of sowing in pre sown experiment while same dosage of insecticides were applied in standing soybean crop during (first fortnight of July). Commutative plant mortality data were recorded after 20, 40, 60 days intervals and compared with untreated control. Total 23 species of white grub beetles recorded on the light trap and Holotrichia serrata Fabricious (Coleoptera: Melolonthinae) was found to be predominant species by recording 20.6% relative abundance out of the total light trap catch (i.e. 1316 beetles) followed by Phyllognathus sp. (14.6% relative abundance). H. rosettae and Heteronychus lioderus occupied third and fourth rank with 11.85% and 9.65% relative abundance, respectively. The emergence of beetles of predominant species started from 15th March, 2018. In April, average light trap catch was 382 white grub beetles, however, peak emergence of most of the white grub species was observed from June to July, 2018 i.e. 336 beetles in June followed by 303 beetles in the July. On the basis of the emergence pattern of white grub beetles, it may be concluded that the Peak Emergence Period (PEP) for the beetles of H. serrata was second fortnight of April for the total period of 15 days. In May, June and July relatively low population of H. serrata was observed. A decreasing trend in light trap catch was observed and went on till September during the study. No single beetle of H. serrata was observed on light trap from September onwards. The cumulative plant mortality data in both the experiments revealed that all the insecticidal treatments were significantly superior in protection-wise (6.49-16.82% cumulative plant mortality) over untreated control where highest plant mortality was 17.28 to 39.65% during study. The mixture of Fipronil 40% and Imidacloprid 40% WG applied at the rate of 300 g a.i. per ha proved to be most effective having lowest plant mortality i.e. 9.29 and 10.94% in pre and post sown crop, followed by Clothianidin 50 WG (120 g a.i. per ha) where the plant mortality was 10.57 and 11.93% in pre and post sown treatments, respectively. Both treatments were found significantly at par among each other. Production-wise, all the insecticidal treatments were found statistically superior (15.00-24.66 q per ha grain yields) over untreated control where the grain yield was 8.25 & 9.13 q per ha. Treatment Fipronil 40% + Imidacloprid 40% WG applied at the rate of 300 g a.i. per ha proved to be most effective and significantly superior over Imidacloprid 70WG applied at the rate of 300 g a.i. per ha.

Keywords: bio efficacy, insecticide, soybean, white grub

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Authors: F. Varga, Z. Liber, J. Jakše, A. Turudić, Z. Šatović, I. Radosavljević, N. Jeran, M. Grdiša

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Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip.; Asteraceae), a source of the commercially dominant plant insecticide pyrethrin, is a species endemic to the eastern Adriatic. Genetic diversity of T. cinerariifolium was previously studied using amplified fragment length polymorphism (AFLP) markers. However, microsatellite markers (single sequence repeats - SSRs) are more informative because they are codominant, highly polymorphic, locus-specific, and more reproducible, and thus are most often used to assess the genetic diversity of plant species. Dalmatian pyrethrum is an outcrossing diploid (2n = 18) whose large genome size and high repeatability have prevented the success of the traditional approach to SSR markers development. The advent of next-generation sequencing combined with the specifically developed method recently enabled the development of, to the author's best knowledge, the first set of SSRs for genomic characterization of Dalmatian pyrethrum, which is essential from the perspective of plant genetic resources conservation. To evaluate the effectiveness of the developed SSR markers in genetic differentiation of Dalmatian pyrethrum populations, a preliminary genetic diversity study was conducted on 30 individuals from three geographically distinct natural populations in Croatia (northern Adriatic island of Mali Lošinj, southern Adriatic island of Čiovo, and Mount Biokovo) based on 12 SSR loci. Analysis of molecular variance (AMOVA) by randomization test with 10,000 permutations was performed in Arlequin 3.5. The average number of alleles per locus, observed and expected heterozygosity, probability of deviations from Hardy-Weinberg equilibrium, and inbreeding coefficient was calculated using GENEPOP 4.4. Genetic distance based on the proportion of common alleles (DPSA) was calculated using MICROSAT. Cluster analysis using the neighbor-joining method with 1,000 bootstraps was performed with PHYLIP to generate a dendrogram. The results of the AMOVA analysis showed that the total SSR diversity was 23% within and 77% between the three populations. A slight deviation from Hardy-Weinberg equilibrium was observed in the Mali Lošinj population. Allele richness ranged from 2.92 to 3.92, with the highest number of private alleles observed in the Mali Lošinj population (17). The average observed DPSA between 30 individuals was 0.557. The highest DPSA (0.875) was observed between several pairs of Dalmatian pyrethrum individuals from the Mali Lošinj and Mt. Biokovo populations, and the lowest between two individuals from the Čiovo population. Neighbor-joining trees, based on DPSA, grouped individuals into clusters according to their population affiliation. The separation of Mt. Biokovo clade was supported (bootstrap value 58%), which is consistent with the previous study on AFLP markers, where isolated populations from Mt. Biokovo differed from the rest of the populations. The developed SSR markers are an effective tool for assessing the genetic diversity and structure of natural Dalmatian pyrethrum populations. These preliminary results are encouraging for a future comprehensive study with a larger sample size across the species' range. Combined with the biochemical data, these highly informative markers could help identify potential genotypes of interest for future development of breeding lines and cultivars that are both resistant to environmental stress and high in pyrethrins. Acknowledgment: This work has been supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.) insecticidal potential’- (PyrDiv) (IP-06-2016-9034) and by project KK.01.1.1.01.0005, Biodiversity and Molecular Plant Breeding, at the Centre of Excellence for Biodiversity and Molecular Plant Breeding (CoE CroP-BioDiv), Zagreb, Croatia.

Keywords: Asteraceae, genetic diversity, genomic SSRs, NGS, pyrethrum, Tanacetum cinerariifolium

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