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Ganoderma boninense Pat. is a persistent soil-borne pathogen that causes significant losses in oil palm (Elaeis guineensis Pat.) productivity. In this study, we evaluated the antifungal activity of volatile organic compounds (VOCs) produced by a fungal isolate, later identified as Paraconiothyrium archidendri F10, against G. boninense. The isolate was identified based on ITS-rDNA sequence analysis and BLASTn results. VOCs produced by P. archidendri F10 were found to inhibit G. boninense mycelium growth by up to 55.8% in four days, with the mycelium exhibiting wavy, non-smooth, and wrinkled morphology, abnormal branching, fused, defective hyphae, and lysis from within. The major VOC components were esters, with 7,9-ditert-butyl-1-oxaspiro[4.5]deca-6,9-diene-2,8-dione being the most abundant (16.72%). The other top-ranking components were 2-O-(6-ethyloctan-3-yl) 1-O-hexyl oxalate (8.71%), methyl heptadecanoate (8.66%), and butyl acetate (5.66%), with minor components comprising less than 5% of the total VOCs. Our findings suggest that P. archidendri F10 has potential as a biofungicide for controlling G. boninense in the field.

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The population response of Heliothis armigera larvae to nuclear polyhedrosis virus (NPV) was investigated. The virus was introduced by permitting the larvae to feed on lettuce leaves. Median lethal doses (LD50) were determined. The estimated LD50 values for the first, second, third, early, and late fourth larval instars were 5, 141, 1226, 5168 and 24553 polyhedra per larva respectively. In the fifth larval instar a degree of maturation resistance against virus infection was observed. An inverse relationship between mortality and larval weight was detected. Expressing the results in terms of LD50 / mg of the larval weight eliminated the observed variation in the susceptibility of larvae. The estimated LD50 values for each of the larval instar groups were used to predict the response of the larval population to virus infection. This procedure provided a sound determination of the response pattern, from which 96% of the variation in the larval susceptibility could be described in terms of the log larval weight.

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Colorado potato beetle, Leptinotarsa decemlineata is one of the major insect pests of potato. Toxicity of spinosad, as a bio-rational insecticide, was investigated against various developmental stages of this pest. Bioassays were conducted by using the eggs, neonates, first, second, third and fourth instar larvae and adults. The potato leaves were impregnated with different concentrations of spinosad and applied for the adults and different larvae bioassays. The eggs were tested through dipping its masses into the insecticide solutions. LC₅₀ values of neonates, first, second, third and fourth instar larvae and unsexed adults after 24 hours were 2.06, 3.19, 4.75, 6.46, 20.24 and 11.97 ppm (of commercial formulation), respectively. Results show that spinosad did not possess any ovicidal effects and the fourth instar larvae and neonates were the most tolerant and the most susceptible stages, respectively. Susceptibility of the neonates (up to 24 hrs after hatching) was significantly higher than that of first instar larvae (24-48 hrs after hatching). Developmental stages of Colorado potato beetle responded differentially to this insecticide. Since the control of L. decemlineata mostly relies on early season measures against the most susceptible stage(s), by considering no ovicidal effect, our results propose a limited interval, for avoiding the highly tolerant larvae.

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This study was conducted to investigate commercial formulations of insecticides against western flower thrips, Frankliniella occidentalis (Pergande).The insects were collected from commercial greenhouses of cucumber in Varamin, Iran. The tested insecticides were diazinon (EC 60%), cypermethrin (EC 40%), fipronil (EC 2.5%), imidacloprid (SC 35%) and a botanical insecticide oxymatrine (Kingbo, AS 0.6%). Fipronil had the highest efficacy among all tested insecticides (LC₅₀ = 17.97 ppm). However, imidacloprid had the lowest efficacy (LC₅₀ = 2303 ppm). The oxymatrine was effective (LC₅₀ = 69.94 ppm) after fipronil.

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The Western flower thrips Frankliniella occidentalis, an important greenhouse pest, has acquired rapid resistance to the chemical pesticides. Therefore, biological control is worth consideration as an alternative control method. Among the biological control agents, entomopathogenic fungi showed to be quite successful in some occasions. In this study, three Iranian isolates of Metarhizium anisopliae (‘DEMI001’, ‘DEMI002’ and ‘DEMI003’) were bioassayed for their lethal effects on the adults of the F. occidentalis, in vitro. The ‘DEMI002’ and ‘DEMI003’ had the lowest and highest LC₅₀ at concentrations of 3.06 ´ 10⁴ and 1.90 ´ 10⁵ conidia/ml, respectively. Also, the isolate ‘DEMI002’ had the lowest LT₅₀ of 4.39 ± 2.13 days at the concentration of 10⁶ conidia/ml. The mean comparison showed that there was a significant difference between DEMI002 and DEMI003 in terms of virulence at most of the concentrations. Consequently, the ‘DEMI002’ can be considered as a promising tool in biological control programs of the F. occidentalis.

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The elm leaf beetle, Xanthogalerucella luteola (Muller) (Col.: Chrysomelidae) is a serious pest of elm trees and it has been distributed all over the world. The current study was undertaken to investigate the inhibitory effects of protein extracts of three weed seeds including datura Datura stramonium L., amaranth Amaranthus retroflexus L. and wild oat Avena fatua L. against X. luteola α-amylase using spectrophotometric assay as well as in gel assays. The effects of five concentrations of each seed proteinaceous extracts were tested on α-amylase activity of the larval gut. The results showed a dose dependent manner in inhibition of the insect enzyme. At the highest concentration of protein extracts (12 μg protein) of all three seed extracts including amaranth, wild oat and datura, the inhibition was 71, 79 and 31%, respectively. Whilst, at low concentration (0.75 μg protein), the inhibition observed was 15, 36 and 5%, respectively. Thus, the greatest inhibition percentage was obtained when proteinaceous extract of wild oat seed was used. These results were confirmed when in gel assays were performed. All three seed proteinaceous extracts had an optimum pH inhibition of 6.0. Thus, it is concluded that wild oat seed proteins are potentially good for detailed investigation in order to get a clear picture of its active compound/s and its structure-function relationship.    

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In this study, gelatin was first extracted by alkaline and acidic treatment including 0.19 N NaOH and 0.12 N acetic acid solution by ratio of skin of rainbow trout (Oncorhynchus mykiss (to solution of 1 to 7 and then heat treatment in 50 °C. Then, hydrolysed by alcalase enzyme for 4 hours with the ratio of enzyme to the substrate 1 to 100 and the degree of hydrolysis were measured after 4 hours. DPPH and ABTS free radical scavenging activity, as well as reducing power assay of gelatin hydrolysate were measured. The results showed that the degree of hydrolysis after 4 hours was 46/7%. Also the highest DPPH and ABTS free radical scavenging and reducing power at concentration of 10 mg/ml were 39/8%, 50/7%, and 0/123, respectively. The skin from fish filleting can be a suitable raw material for extraction of peptides with biological activities. The results showed that peptides derived from rainbow trout fish skin gelatin can be considered as a natural antioxidant.

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 Fish protein hydrolysates from whole kilka, using alcalase enzyme (ratio 1: 100) under optimal temperature (55°C) and pH (8.5) was evaluated for its hydrolysis degree and antioxidant activity. Results of the hydrolysis degree recorded at time intervals of 1, 2, 3 and 4 hours indicated the hydrolysis degree increased with increase in the hydrolysis time. The evaluation of FPH antioxidant activity, using DPPH, ABTS and reducing power assay tests at 3 concentrations (1, 2 and 5 mg/ml indicated the highest inhibitory effect at 5 mg/ml was 74.4%, 72.3% and 1.8 absorbance in 700 nm for DPPH, ABTS and reducing power assay, respectively. Generally, the findings of this research confirmed the potential of kilka as a source of natural antioxidants for food applications.

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Background: The outbreak of novel coronavirus (2019-nCoV), which began in Wuhan, China, has rapidly spread in many countries and is currently considered a pandemic. The virus (SARS-CoV-2) causes severe acute respiratory syndrome and is related to SARS-CoV and MERS-CoV.
Methods: In this review, an introduction to SARS-CoV-2 was provided comprising the following items: general features; pathogenesis; the existing knowledge on immunological properties; transmission routs; diagnostic features, especially discussion about new approaches for treatment and prevention; and different diagnostic methods including nucleic acid based assays, serological testing, and MALDI TOF-MS and LC-MS technologies.
Findings and Conclusion: Introducing the different methods for SARS-CoV-2 detection may be useful to provide new insights into the development and improvement of detection primers, probes, methods/techniques, potential targets for drug designation, and therapeutic candidates against the virus.

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Acute toxicity of the field recommended concentration of three conventional insecticides (Diazinon, Malathion and Chlorpyrifos) and mineral oil was evaluated on 3rd and 4th instar larvae and adults of Cryptolaemus montrouzieri Mulsant. The mortalities caused by the insecticides and mineral oil were significantly different. Diazinon and Malathion with 100% mortality showed the highest toxicity to the different stages of the ladybird. Chlorpyrifos and mineral oil caused less than 30% mortality, while mineral oil had the lowest harmful effect on this predator. Based on LC50 and LC90 values 24h after treatment, the male and female adults of C. montrouzieri were more susceptible to Diazinon (701 and 635; 1257 and 1194ppm) than to Chlorpyrifos (4238 and 4316, 5683 and 5480 ppm). Based on International Organization of Biological Control (IOBC) classification, Chlorpyrifos and mineral oil were classified as category 1 (harmless) and Diazinon and Malathion were placed in category 4 (harmful).

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The potato tuber moth (PTM), Phthorimaea operculella (Zeller), is a major pest of potato, both in the field and storehouses. In this study, we have evaluated the lethal effects and persistence of Zingiber officinale (Roscoe) pure (PEO) and nano-formulated essential oil (NFO) on different developmental stages (egg, male and female adults) of PTM. Essential oil was extracted by hydro-distillation using a Clevenger-type apparatus. The essential oil was analyzed by gas chromatography/mass spectrometry (GC/MS). Nanofibers were produced by electrospinning technique. The morphology of nanofibers was investigated by SEM. Fourier transform infrared (FTIR) was used to identify the characteristic functional groups in the PEO, nanofiber and PEO/NFO scaffold. Bioassays were performed in 250 ml glass jars. The essential oil consisted of α-Zingiberene as the most abundant component (14.21%), followed by Ar-curcumene (12.58%), β-sesquiphellandrene (12.48%) and cis-α-bisabolene (10.29%). The results of FTIR spectra showed the establishment of the functional groups of PEO on the structure of the nanofiber. The images of SEM also demonstrated the establishment of PEO in the structure of the nanofiber. LC50 values of PEO and NFO were estimated 75.44 and 30.24µl/l air for eggs, 19.08 and 10.28µl/l air for female adults, and 17.76 and 9.56µl/l air for male adults, respectively. Persistence data showed that nano-formulated essential oil (49 days) in comparison with pure essential oil (15 day) had longer persistence. The results demonstrated that Z. officinale PEO and its nano-formulation could play an important role as natural pesticides for the management of PTM.

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The Cu dependent restriction deoxyribozyme is the unique example of known deoxyribozymes. The uniqueness of this deoxyribozyme is originated from specific cleaving of single stranded DNA and formation of triple helix DNA structure which is necessary for substrate recognition and binding. The most established method for measuring the kinetic parameters of deoxyribozyme is based on use of radiolabeled substrates which have several difficulties. In this study we present accurate, fast and inexpensive methods for kinetic study of the deoxyribozyme which is based on extrinsic fluorescence and UV-visible spectroscopy techniques. As mentioned above, DNA triple helix formation is necessary for substrate identification and also enzyme activity. Circular dichroism spetropolarimetery is used for structural study of enzyme. Analysis of spectrum results from this technique indicates structural changes which is a direct evidence for the triple helix formation in enzyme-substrate complex. Extrinsic fluorescence experiment is based on high affinity of SYBR gold to double stranded DNA compared to single stranded DNA. Enzyme activity can be measured by SYBR gold fluorescence emission upon addition of cofactor to the enzyme-substrate complex. Continuous hyperchromicity assay method which is based on UV-visible spectroscopy was used for measuring of enzyme activity by hyperchromicity of the enzyme-substrate complex after addition of cofactor. Comparison of the results show that the continuous hyperchromicity assay is more accurate than the extrinsic fluorescence method, because of this method is based on intrinsic physicochemical properties of DNA without interference of external factors.

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Backgrounds: This study aimed to investigate the prevalence of cryptococcaemia in HIV infected patients with CD4 counts of ≤100 cells/mm3 in a tertiary care hospital.
Materials & Methods: The present cross sectional study was conducted at the Sri Guru Ram Das Institute of Health Sciences and Research, India, as a tertiary care hospital. All HIV infected patients with CD4 counts of ≤100 cells/mm3, referring to the hospital during May 2020 to May 2021 were enrolled in this study. Blood samples taken from patients were processed for wet mounting, negative staining with India ink, gram staining, fungal culture, and cryptococcal antigen (CrAg) lateral flow assay (LFA). Statistical analysis was done using SPSS software Version 20.0 (SPSS Inc. Chicago, IL, USA) by employing Chi-square and Fisher’s exact tests to compare categorical variables.
Findings: Out of 100 patients enrolled, 28 (28%) cases had CD4 counts below 50, while 72 (84.7%) patients had CD4 counts in the range of 51-100. Also, 55 patients (55%) received antiretroviral therapy (ART), and 45 (45%) cases were ART naï􀇅ve. About 56% of patients had no opportunistic infections, and 37% had pulmonary tuberculosis. Three samples were positive in LFA, showing a prevalence of 3%, while only one of the culture samples was positive for Cryptococcus species. However, low CD4 count was found to be strongly correlated with positive serum cryptococcal antigenemia.
Conclusion: The present study reveals that cryptococcal antigenemia is a health problem, and that cryptococcal antigen screening and treatment policy recommended by WHO should be performed routinely for HIV patients registered in ART centres in the current setting, especially for those who are ART naï􀇅ve and have CD4 counts of ≤100 cells/mm3.

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Current investigations indicate different in vitro infection patterns for Oka and Dumas strains. Infection of cells in culture with Dumas strain produces lower number of infectious particles while the Oka strain is highly infectious in vitro. It is postulated that weak expression of the replication genes in Dumas strain may be the reason for the attenuated phenotype. The objective of this study was to analyze the sequences of the promoter region in two of the VZV replication genes, 16 and 52 by studying the level of expression of a reporter gene. For this purpose, primers were designed from VZV published sequences to amplify the promoter regions of both genes using Polymerase Chain Reaction (PCR) technique. The amplicons were cloned in a lacZ reporter vector. Cotransfaction of 52-Oka reporter plasmid with virus major trans-activator (IE62) in Huh7 cells showed that the presence of 52-Oka promoter was up regulated by the IE 62 trans-activator in a dose dependent manner resulting in ß-gal levels approximately 4-fold higher than those observed with 52-Dumas promoter and 10-fold higher than basal levels. In addition cotransfection of 16-Oka reporter plasmid did not show any significant change in activity in comparison with 16-Domus-reporter plasmid. Sequence determination of the promoter region in gene 52 indicated differences in 3 nucleotides in Dumas strain compared to Oka strain while no change was observed in the promoter sequences of gene 16 of the two strains. It is hence postulated a relationship between mutations in the Dumas promoter of replication genes, and the lower infectivity of the Dumas strain.

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The two-spotted spider mite, Tetranychus urticae (Acari: Tetranychidae) is one of the most important pests of field crops, orchard trees and ornamentals around the world. The short life cycle, high reproductive potential, accompanied by frequent acaricide applications have caused resistance development to wide range of acaricides. In this study the susceptibility of two populations, collected from Karaj and Mahllat, was investigated against fenpyroximate. The bioassay test was carried out by using the leaf-dip method. The results showed that the LC₅₀ values for Karaj and Mahallat population were 2.1 and 92 (mg/ml), respectively. The resistance ratio was 43.8. The enzyme assay results revealed that the activity ratios of esterase in Mahallat to Karaj populations were 2.5 and 1.2 when α-NA and β-NA were used as a substrate, respectively. The activity of cytochrome P₄₅₀ in Mahallat population was 1.37 times higher than the Karaj population. There was no significant difference in glutathion S-transferase activity between the two populations. The gene expression (qRT-PCR) results showed that the expression level of CYP392A11 in Mahallat population was 3.52 times higher than Karaj population. These results suggested that esterase and cytochrome P₄₅₀ monoxygenase are probably involved in resistance of T. urticae to fenpyroximate. 

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Objective: Bacterial meningitis is a dangerous and sometimes fatal infection that affects the central nervous system. Because some antibiotics can prevent some types of these Bacteria and supress them from spreading and infecting, therefore it is important to know what type of virus or bacterium is causing meningitis. Haemophilus influenzae and Neisseria meningitides are the two main pathogens causinig acute bacterial meningitis. Different methods are used for the detection of H. influenzae and N. meningitidis but they are of low sensitivity, taking long time and difficult to perform. Therefore, complementary methods are necessary for more sensitive detection of these agents. Materials and Methods: In this study, a multiplex polymerase chain reaction (mPCR) assay was developed for detection of H. influenzae and N. meningitidis. These strains were confirmed by biochemical methods. Two specific primer pairs were designed for lic-1 and opa genes of H. influenzae and N. meningitidis respectively. Results: DNA amplification product fragments were 150 bp and 320 bp for H. influenzae and N. meningitidis, respectively. Streptococcus pneumoniae used as a negative control and did not yield a PCR product. Conclusion: The results of this study indicated that PCR is a useful complementary diagnostic technique, especially when Gram stain, culture, or antigenic detection is negative or inconclusive.