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Genus Pinus is a widely dispersed genus of conifer plants in the Northern Hemisphere. However, the inadequate accessibility of genomic knowledge limits our understanding of molecular phylogeny and evolution of Pinus species. In this study, the evolutionary features of complete plastid genome and the phylogeny of the Pinus genus were studied. A total of thirteen divergent hotspot regions (trnk-UUU, matK, trnQ-UUG, atpF, atpH, rpoC1, rpoC2, rpoB, ycf2, ycf1, trnD-GUC, trnY-GUA, and trnH-GUG) were identified that would be utilized as possible genetic markers for determination of phylogeny and population genetics analysis of Pinus species. Furthermore, seven genes (petD, psaI, psaM, matK, rps18, ycf1, and ycf2) with positive selection site in Pinus species were identified. Based on the whole genome this phylogenetic study showed that twenty-four Pinus species form a significant genealogical clade. Divergence time showed that the Pinus species originated about 100 million years ago (MYA) (95% HPD, 101.76.35–109.79 MYA), in lateral stages of Cretaceous. Moreover, two of the subgenera are consequently originated in 85.05 MYA (95% HPD, 81.04–88.02 MYA). This study provides a phylogenetic relationship and a chronological framework for the future study of the molecular evolution of the Pinus species.

Coronavirus disease 2019 (COVID-19) has caused mild illness in children, until the emergence of the novel hyperinflammatory condition paediatric inflammatory multisystem syndrome temporally associated with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) (PIMS-TS). PIMS-TS is thought to be a post-SARS-CoV-2 immune dysregulation with excessive inflammatory cytokine release. We studied 25 hydroxyvitamin D (25OHD) concentrations in children with PIMS-TS, admitted to a tertiary paediatric hospital in the UK, due to its postulated role in cytokine regulation and immune response. Eighteen children (median (range) age 8·9 (0·3-14·6) years, male = 10) met the case definition. The majority were of Black, Asian and Minority Ethnic (BAME) origin (89 %, 16/18). Positive SARS-CoV-2 IgG antibodies were present in 94 % (17/18) and RNA by PCR in 6 % (1/18). Seventy-eight percentage of the cohort were vitamin D deficient (< 30 nmol/l). The mean 25OHD concentration was significantly lower when compared with the population mean from the 2015/16 National Diet and Nutrition Survey (children aged 4–10 years) (24 v. 54 nmol/l (95 % CI −38·6, −19·7); P < 0·001). The paediatric intensive care unit (PICU) group had lower mean 25OHD concentrations compared with the non-PICU group, but this was not statistically significant (19·5 v. 31·9 nmol/l; P = 0·11). The higher susceptibility of BAME children to PIMS-TS and also vitamin D deficiency merits contemplation. Whilst any link between vitamin D deficiency and the severity of COVID-19 and related conditions including PIMS-TS requires further evidence, public health measures to improve vitamin D status of the UK BAME population have been long overdue.

Several studies demonstrated that in insects cuticle melanism is interrelated with pathogen resistance, as melanin‐based coloration and innate immunity possess similar physiological pathways. For some insects, higher pathogen resistance was observed in darker individuals than in individuals with lighter cuticular coloration. Here, we investigated the difference in immune response between two color morphs (black and red) and between the life stages (pupa and adult) of the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae). Here in this study, cuticle thickness, microbial test (antimicrobial activity, phenoloxidase activity, and hemocyte density), and immune‐related gene expression were evaluated at different stages of RPW. Study results revealed that cuticle thickness of black phenotype was thicker than red phenotype at old‐pupa stage, while no significant difference found at adult stage. These results may relate to the development processes of epidermis in different stages of RPW. The results of antimicrobial activity, phenoloxidase (PO) activity, and hemocyte density analyses showed that adults with a red phenotype had stronger pathogen resistance than those with a black phenotype. In addition to antimicrobial activity and PO activity, we tested relative gene expression in the fat body of old pupae. The results of hemolymph antimicrobial analysis showed that old pupae with a red phenotype were significantly different from those with a black phenotype at 12 hr after Staphylococcus aureus injection, suggesting that red phenotype pupae were more sensitive to S. aureus. Examination of gene expression in the fat body also revealed that the red phenotype had a higher immune response than the black phenotype. Our results were inconsistent with the previous conclusion that dark insects had increased immune function, suggesting that the relationship between cuticle pigmentation and immune function in insects was not a direct link. Additional possible factors that are associated with the immune response, such as life‐history, developmental, physiological factors also need to be considered., This paper reported pathogen resistance differences between two different segmental ventral colorations in the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae) (red phenotype and black phenotype) and investigated immune function in the old‐pupa and adult stages. Our results showed cuticle melanism pigmentation does not correspond to enhanced immune function in RPW, which were inconsistent with the previous conclusion that dark insects had increased immune function. We suggest that evaluation of the relationship between cuticle pigmentation and immune function in insects needs to also consider additional possible factors that are associated with the immune response, such as life‐history, developmental, and physiological factors.

The use of pesticides may pose a serious threat to soil and water quality, human health, food safety, aquatic species and beneficial insects. Biological control is an effective component of integrated pest management comprising importation, augmentation and conservation. Conservation of natural enemies could enhance or limit the efficiency of herbivorous insect pests. The present study was designed to evaluate the efficiency of Diaeretiella rapae (McIntosh) and other food resources impacts on parasitoid’s fecundity, survival and efficiency. The effect of different insecticides [Carbosulfan, Imidacloprid, Diafenthiuron and Lufenuron] was also evaluated in a laboratory controlled experiment. The results revealed that females fed on aphid honeydew proved to be of poor quality as compared to other sugar sources, including honey and sugar solution and has a positive impact of mating on monogenesis. The negative impact was observed on oviposition activity by female reared in the laboratory having less number of mature eggs than females collected in the field. All insecticides demonstrated dose-dependent mortality of D. rapae. In conclusion, food and host availability and insecticides affect the efficiency of D. rapae. These results could lead to the hypothesis that D. rapae females have a high mortality rate in the field due to lower aphid hosts.

Understanding circadian foraging rhythms activity of the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae) foragers at different temperatures is an important step towards developing control measures in Integrated Pest Management (IPM) programs. In this study, the circadian foraging rhythm activities of S. invicta foragersat different temperature were investigated under laboratory and field conditions. Results indicated that the foraging activity increased after sunrise, and maximum foraging occurred at 14:00 (foraging rate was 69.22 ± 0.57 and 72.58 ± 1.15 foragers/min in the first and second year, respectively) in the tea fields of Guangzhou during autumn. Furthermore, foragers demonstrated circadian rhythms and exhibited a unimodal after 24 h. A significant correlation was found between foraging activity and temperature. S. invicta colonies were active at moderate soil temperatures (approximately 26.65 °C to 29.24 °C). The preferred temperature of the colonies was 26 °C, followed by 22 °C and 18 °C in the laboratory. The individual S. invicta activity was maximum at 17:00 (18.67 ± 1.66 times /10 min) and minimum at 5:00 (8.33 ± 2.51 times/10 min) at 26 °C. The fluctuating temperature had a significant impact on individual locomotor activity (r = 0.8979, P

Worldwide, stored products are attacked by a large number of pests resulting in significant economic losses. Among these stored grain pests, khapra beetle, Trogoderma granarium (Everts) (Coleoptera: Dermestidae) is one of the top ranked pests that has ability to survive under diverse climate conditions. The management of this pest is mainly done by using synthetic chemicals which have side effects on consumers, ecosystem and non-target organisms. However, phyto-derivatives were found to be the effective environment friendly alternatives against T. granarium. Therefore, in this review, success of phyto-derivatives against khapra beetle from conventional means to modern research has been critically analyzed, summarized and discussed. In context, the different life stages of the insect starting from egg laying to adult development have been briefly explained. The review focuses upon recent research conducted on the evaluation of dozens of phyto-derivatives. In addition, the article has also highlighted some limitations of plant derived compounds and concludes via hoping that the future formulated pesticides will be safer, economical, least toxic to human and our planet ecosystem.

Whitefly, Bemisia tabaci (Hemiptera: Aleyrodidae), is a substantial pest of Okra crop, Abelmoschus esculentus, that caused substantial losses through sucking cell sap. On the basis of susceptibility, a field experiment was carried out to assess the different genotypes of A. esculentus during growing season (July–August) against B. tabaci. Plant genotype resistance and vigor were also evaluated in this study. Meanwhile, morphological parameters were also recorded as well as whitefly-infested and healthy leaves were collected for biochemical analysis. The obtained results revealed that the Okra varieties A. esculentus OK-1304 and Pen Beauty were less infested (mean populations 4.10, 4.97 adults/leaf, respectively) while OK-1307 was highly susceptible to B. tabaci (10.22 adults/leaf). Meanwhile, comparative analysis revealed that B. tabaci population was negatively correlated to relative humidity and positively correlated with low and high temperature. The low concentrations (0.5%) of salicylic acid and citric acid effectively reduced the infestation to 4.63 adults/leaf and 4.87 adult/leaf, respectively, as compared to control (10.17 adult/leaf). The biochemical analysis represented that the catalase concentration decreased in infested leaves and the concentrations of peroxidase, phenolics and superoxide dismutase that act as resistant compound against insect feeding increased.

Panonychus citri (citrus red mite) is a devastating pest of citrus orchards. The conventional chemical acaricides have been strongly forbidden for the management of agricultural insect pests in China. Therefore, we evaluated the susceptibility of adult and nymphs P. citri in laboratory against eight isolates of four fungal species, Akanthomyces lecanii, Metarhizium anisopliae, Beauveria bassiana and Aschersonia aleyrodis. Each citrus seedling having 40 adults (2-d-old) and nymphs (on separate plants) were sprayed with isolates at the concentration of 104 ~ 108 conidia mLˉ1 whereas controlled seedlings were sprayed with 0.02% Tween-80. After 9 days of fungal exposure, the four fungal isolates caused more than 50% mortality of mites, such as; 85.6%, 87.9%, 64.6% and 79.7% by A. lecanii (V3450), B. bassiana (BFZ0409), M. anisopliae (MFZ0706) and A. aleyrodis (AsG0910), respectively. The nymphal mites were less susceptible to applied fungi compared to adults. The LC50s of the tested isolates were determined by the fitted time-concentration-mortality relationships, which declined over days after spray. LT50s were decreased with a high concentration of isolates. After the 9-d inoculation, two isolates of B. bassiana (BFZ0409 and D1344) and one isolate of A. lecanii (V3450) were highly effective at the minimal dose of LC50 of 104 conidia mLˉ1 and are promising candidates to control mites, as compared to other tested fungal isolates.

Hybrid seeds of several important crops with supreme qualities including yield, biotic and abiotic stress tolerance have been cultivated for decades. Thus far, a major challenge with hybrid seeds is that they do not have the ability to produce plants with the same qualities over subsequent generations. Apomixis, an asexual mode of reproduction by avoiding meiosis, exists naturally in flowering plants, and ultimately leads to seed production. Apomixis has the potential to preserve hybrid vigor for multiple generations in economically important plant genotypes. The evolution and genetics of asexual seed production are unclear, and much more effort will be required to determine the genetic architecture of this phenomenon. To fix hybrid vigor, synthetic apomixis has been suggested. The development of MiMe (mitosis instead of meiosis) genotypes has been utilized for clonal gamete production. However, the identification and parental origin of genes responsible for synthetic apomixis are little known and need further clarification. Genome modifications utilizing genome editing technologies (GETs), such as clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (cas), a reverse genetics tool, have paved the way toward the utilization of emerging technologies in plant molecular biology. Over the last decade, several genes in important crops have been successfully edited. The vast availability of GETs has made functional genomics studies easy to conduct in crops important for food security. Disruption in the expression of genes specific to egg cell MATRILINEAL (MTL) through the CRISPR/Cas genome editing system promotes the induction of haploid seed, whereas triple knockout of the Baby Boom (BBM) genes BBM1, BBM2, and BBM3 cause embryo arrest and abortion, which can be fully rescued by male-transmitted BBM1. The establishment of synthetic apomixis by engineering the MiMe genotype by genome editing of BBM1 expression or disruption of MTL leads to clonal seed production and heritability for multiple generations. In the present review, we discuss current developments related to the use of CRISPR/Cas technology in plants and the possibility of promoting apomixis in crops to preserve hybrid vigor. In addition, genetics, evolution, epigenetic modifications, and strategies for MiMe genotype development are discussed in detail.

Root-knot nematodes Meloidogyne incognita (Kofoid and White) Chitwood and Rhizoctonia bataticola (Taub.) Butler, fungus, are very dangerous root damaging pathogens. Present study was planned to establish a chemical control of these root deteriorating pathogens under lab conditions as well as in field. Maximum death rate of nematode juveniles and minimum numbers of nematode eggs hatched were recorded in plates treated with Cadusafos (Rugby® 100G) @12 g/100 ml and Cartap® (4% G) @9g/100 ml. Chemical treatment of Rhizoctonia bataticola with Trifloxystrobin + Tebuconazole (Nativo®) @0.2 g/100 ml and Mancozeb + Matalaxyl (Axiom) @0.25 g/100 ml significantly controlled the mycelial growth in plates. The best treatments tested in laboratory were applied in field as protective and curative treatments. Results proved that chemical control of root-knot nematode and root rot fungi by tested chemicals at recommended time and dose is a significant management technique under field conditions.

The lepidopteran insect pests have significant importance in vegetable production. The present study was performed to investigate the baseline studies about the assessment of feeding and consumption potential, utilization indices and losses promises of leafworm, Spodoptera litura (Fab.) on Okra. The data regarding feeding potential, food utilization and consumption indices as well as losses of different larval instars were recorded and subjected to appropriate statistical analysis. The results showed that, in the beginning, the approximate digestibility of various instars was increase, e.g. third instar (51.36%–64.03%), fourth instar (63.42%–69.45%) and fifth instar (70.25%–76.10%). However, after a certain period, the digestibility was decreased and efficiency to convert the ingested food into biomass varied significantly. The consumption index values increased with an increase in time but the consumption and growth rate was declined of fourth instar larvae. The ingestion and digestion increased of third (10.01–13.06, 8.32–11.91 mg), fourth (11.27–17.28, 10.96–14.03 mg) and fifth (12.60–19.40, 11.93–15.28 mg) larval instars. The corrected weight of consumed leaves increased with a gain in body weight. However, in the third instar, a decline was observed on the last day of feeding. Maximum leaf area was consumed by fifth instar larvae (44.66 cm2) followed by fourth (35.41 cm2) and third (27.98 cm2) instars. In conclusion, all the dependent parameters, including food utilization potential, consumption indices and losses were higher for fifth instar larvae than others. These results emphasized the re-establishment of fundamental (economic threshold level: ETL, economic injury level: EIL) integrated pest management concepts.

Whiteflies, Bemisia tabaci Gennadius (Aleyrodidae: Hemiptera), are a polyphagous economically destructive pest of several solanaceous crops around the world. Many secondary metabolites are synthesized by different biotrophic and necrotrophic fungi which are capable of inducing systemic resistance in plants against various phytophagous pests. The present laboratory work demonstrated the anti-insect impact of a protein extracted and purified partially from an entomopathogenic fungus (EPF) Lecanicillium lecanii (Zimmermann) against B. tabaci. Three different concentrations (i.e., 7.43, 11.15, and 22.31 μg mL−1) of this protein were bioassayed to assess its effect on the fecundity rate of B. tabaci on cotton (Gossypium hirsutum L.) plants. Furthermore, the possible implication of this fungal protein in defense pathways of cotton plants was evaluated by determining the expression profiles of salicylic acid (SA) and jasmonic acid (JA) pathways related to major genes through reverse transcription qPCR (RT-qPCR). According to the results, all protein concentrations exerted a significant (F3, 252 = 62.51, p ≤ 0.001) and negative impact on the fecundity rate of B. tabaci females. At the highest protein concentration (22.31 μg mL−1), the minimum rate of fecundity (i.e., 2.46 eggs female−1day−1) of B. tabaci was noted on the seventh day, whereas fecundity rates for the other two protein concentrations (i.e., 11.15 and 7.43 μg mL−1) were, respectively, 3.06 and 3.90 eggs day−1 female−1. The maximum rate of fecundity (6.01 eggs female−1day−1) was recorded in untreated (control) treatments. In addition, the foliar application of L. lecanii derived protein significantly upregulated all SA linked genes (OPR3, PPO1 and COI1) and slightly triggered up the JA linked genes (LOX1, UBQ7 and AOS) in the cotton plants. These findings revealed that this L. lecanii extracted partially purified protein triggered systemic resistance against B. tabaci in the cotton plants, proposing its putative effectiveness as an innovative biological control technique against B. tabaci and other phloem-feeding hemipteran pests. Nevertheless, further investigations such as purification and molecular and functional characterization of this L. lecanii-derived partially purified protein are required.

As an antioxidant, alpha-tocopherol (α-Toc) protects plants from salinity-induced oxidative bursts. This study was conducted twice to determine the effect of α-Toc as a foliar spray (at 0 (no spray), 100, 200, and 300 mg L−1) to improve the yield and biochemical constituents of fresh green capsules of okra (Abelmoschus esculentus L. Moench) under salt stress (0 and 100 mM). Salt stress significantly reduced K+ and Ca2+ ion concentration and yield, whereas it increased H2O2, malondialdehyde (MDA), Na+, glycine betaine (GB), total free proline, total phenolics, and the activities of catalase (CAT), guaiacol peroxidase (GPX), and protease in both okra varieties (Noori and Sabzpari). Foliar application of α-Toc significantly improved the yield in tested okra varieties by increasing the activity of antioxidants (CAT, GPX, SOD, and ascorbic acid), accumulation of GB, and total free proline in fruit tissues under saline and non-saline conditions. Moreover, α-Toc application as a foliar spray alleviated the adverse effects of salt stress by reducing Na+ concentration, MDA, and H2O2 levels and improving the uptake of K+ and Ca2+. Among the tested okra varieties, Noori performed better than Sabzpari across all physio-biochemical attributes. Of all the foliar-applied α-Toc levels, 200 mg L−1 and 300 mg L−1 were more effective in the amelioration of salinity-induced adverse effects in okra. Thus, we concluded that higher levels of α-Toc (200 mg L−1 and 300 mg L−1) combat salinity stress more effectively by boosting the antioxidant potential of okra plants.

The experiment was conducted during summer seasons (2014), in Demonstration Farm of the Collage of Agricultural Studies at Shambat, Sudan University of Science and Technology, in order to study the effect of phosphorus fertilizer on growth and productivity of five cultivars of sorghum (Sorghum bicolor L. (Moench)., and also to evaluate phosphorus use efficiency. The experiment was arranged in split plot trial with addition of phosphorus as main plot (with and without) and five sorghum cultivars (Butana, Tabat, Arfagadamak, Wad Ahmed, Tetron) as the sub-plot with four replications. Plant height(cm), number Leaves, Leaf area(cm2), Stem diameter plant fresh weight(kg), plant dry weight(kg), length of panicle(cm), weight of seeds panicle(g), weight of 100 seeds(g), number of panicle, yied/t/ha and phosphorus use efficiency, were measured. The results revealed that phosphorus affected yield and growth characters. Arfagadamak cultivar revealed the best use, therefore the highest productivity (12.75 tan/h), and Wadahmed showed the lowest use of phosphorus and therefore the lowest productivity (9.45 tan/h).

Noncoding RNAs play essential functions during epigenetic regulation of gene expression and development in numerous organisms. Three type of small noncoding RNAs found in eukaryotes, which are small interfering RNAs (siRNAs), microRNAs (miRNAs) and piwi-interacting RNAs (piRNAs). Small RNAs (sRNAs) originated from infecting viruses are known as virus-derived small interfering RNAs (vsiRNAs), are responsible for RNA silencing in plants. However, Virus-induced gene silencing (VIGS) is mainly dependent on RNA silencing (RNAi). Interestingly, RNA silencing happens in plants and insects during viral infections. VsiRNAs originate from dsRNA molecules which further require hosts Dicer-like (DCL) proteins, RNA dependent RNA polymerase (RdRP) proteins, and Argonaute (AGO) proteins. RdRP uses ssRNA for complete RNA amplification process as well as DCL dependent secondary vsiRNA formation. Viral Suppressors of RNA silencing (VSRs) interfere with the movement of signals during silencing mechanism. Moreover, intercellular movement of viruses is facilitated by virus-encoded movement proteins. Proteomic and Transcriptomic mechanisms regulated by specific factors like microRNAs, which has become an essential factor of gene regulation. RNAi is also involved in gene suppression by regulating the transcriptional and post-transcriptional gene expression in many eukaryotes. Rice grassy stunt virus (RGSV) is a member of genus Tenuivirus. Although, there is no much work done on RGSV, but this virus has become very potent and destructive, and effects rice crop in many Asian countries, particularly in China. In this review, we have highlighted the rice viruses' biology and silencing suppressors. This work will be helpful for plant virologists in understanding the role of vsiRNAs mechanism in rice viruses especially RGSV.

Global food security is threatened by insect pests of economically important crops. Chemical pesticides have been used frequently for the last few decades to manage insect pests throughout the world. However, these chemicals are hazardous for human health as well as the ecosystem. In addition, several pests have evolved resistance to many chemicals. Finding environment friendly alternatives lead the researchers to introduce biocontrol agents such as entomopathogenic fungi (EPF). These fungi include various genera that can infect and kill insects efficiently. Moreover, EPFs have considerable host specificity with a mild effect on non-target organisms and can be produced in bulk quantity quickly. However, insights into the biology of EPF and mechanism of action are of prime significance for their efficient utilization as a biocontrol agent. This review focuses on EPF-mediated insect management by explaining particular EPF strains and their general mode of action. We have comprehensively discussed which criteria should be used for the selection of pertinent EPF, and which aspects can impact the EPF efficiency. Finally, we have outlined various advantages of EPF and their limitations. The article summarizes the prospects related to EPF utilization as biocontrol agents. We hope that future strategies for the management of insects will be safer for our planet.

Cotton holds immense economic potential owing to its premium quality fiber, oil, and protein contents. The ever-growing human population along with a global shift in climate requires a significant upsurge in cotton production. In the past years, conventional breeding tools were utilized to improve yield and quality through introgression of genes in elite cotton cultivars. The conventional breeding methods hold several limitations especially time-consuming, reliance on naturally available genetic variations, and extensive backcrossing. The modern plant breeding approaches especially modern genome editing technologies (GETs) can be utilized to achieve objectives within a limited time. The GETs including zinc-finger nucleases, transcription-activator-like effector nucleases, and clustered regularly interspaced palindromic repeats and CRISPR-associated proteins systems (CRISPR/Cas)-based technologies have been exploited in various crop improvement programs. The CRISPR/Cas system possesses immense potential owing to simplicity, competency, and versatility which lacks in other GETs. In cotton, the CRISPR/Cas system can help to enhance biotic and abiotic stress resistance, modify gene expression, and gene stacking of important traits with minimum chances of segregation. The transgene clean approach further enhances CRISPR acceptability, and plants can further utilized for selfing or backcrossing to improve traits under investigation. In this chapter, we highlight the avenues of research to utilize CRISPR/Cas system to combat biotic and abiotic stresses, fiber quality, plant architecture and flowering, epigenetic modifications, and gene stacking for economically important traits. Furthermore, the challenges associated with CRISPR application in cotton and its future prospects are addressed in detail.

Background Pink bollworm, Pectinophora gossypiella (Lepidoptera: Gelechiidae) is a native pest of Asia and preferably invasion on cotton (Gossypium hirsutum L.) crop as a commendatory host plant. Commercially, G. hirsutum is known as white gold and is an important cash crop all over the globe. Limited studies were published to focus on certain dietary compositions against different cotton pests. Therefore, the present study was undertaken in the laboratory under controlled conditions (temperature: 27 ± 2°C and relative humidity: 60 ± 10%) to determine the impact of three different treatment diets (wheat germ meal, okra, and chickpea) on the biological aspects (lifetime, developmental period) of P. gossypiella. Results Results revealed that the shortest larval time of P. gossypiella was observed on the okra feed diet while the longest period was recorded on the wheat germ diet. Meanwhile, the pupation delay was noted on the wheat germ diet. The dietary influence was also observed on adult stages of female and male P. gossypiella (43.00 and 37.50 days respectively) and compared with a standard diet (56.50 and 52.50 days respectively). Furthermore, larval weighed more on the okra and chickpea diet followed by the wheat germ diet, whereas highest pupal weight was observed on the standard diet followed by the chickpea diet and okra diet. Conclusion Developmental parameters were significantly variant across all treatment diets, whereas the higher significant difference was reported on the okra diet. Therefore, the existing data of this study offers fruitful interventions for the future as a modified diet for large-scale and rapid mass production of P. gossypiella larvae.

The new species of a ladybird beetle, Cryptogonus bhalwalnensis Azad & Wang sp. nov. is described from Pakistan. Cryptogonus nepalensis bhutanensis Bielawski is recorded for the first time from Pakistan. Both species of Cryptogonus from Pakistan are diagnosed and illustrated; a distribution map is also presented.

Background Whitefly, Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae), is one of the most harmful pests in greenhouses and in open fields. Main body The present study aimed to assess in vitro virulence of 3 entomopathogenic fungal strains (EPFs) of Lecanicillium lecanii 3 (V-3), 4 (V-4), and 5 (V-5) against the whitefly, Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae), by spray method. The 3 disparate bioassays were performed encompassed of conidial concentrations and fungal filtrate of the strains, V-3, V-4, and V-5, and also their binary combinations. In the filtrate bioassay, 2 ml of fungal filtrate of each strain was used. In the conidial bioassay, 3 disparate concentrations (1 × 105, 1 × 106, and 1 × 107 conidia ml−1) were used for each fungal strain, while in the binary combinations (1 ml filtrate + 1 ml conidia) of V-3 × V-3, V-4 × V-4, and V-5 × V-5 were used. Mortality rates against the whitefly were recorded on the 7th day. In the conidial bioassay, maximum mortality rates were found at V-3 strain (90.6%), V-4 strain (78.4%), and V-5 strain (83.6%) at the highest concentration (1 × 107 conidia ml−1) on the 7th day. In the filtrate bioassay, V-3 strain revealed a maximum mortality (93%), V-4 strain (85%), and then V-5 strain (87%) on the 7th day. Moreover, in the bioassay of binary combinations, the highest mortality rate of the whitefly was counted in V-3 × V-3 strain (84.6%), V-4 × V-4 strain (70.6%), and V-5 × V-5 strain (79.8%) on the 7th day. Conclusion All treatments had the potential to control B. tabaci significantly. In all bioassays, the V-3 strain was the extreme virulent, and the filtrate application of V-3 strain was the utmost impressive against B. tabaci.

The present study was performed to assess the individual and combined potential of 3 different concentrations of Trichoderma sp. (Ts) and Bacillus thuringiensis (Bt) to control brinjal insect pests. Tested formulations were applied on larval and adult stages of the pest. The Trichoderma laboratory bioassays revealed 73% mortality of the aphid species, Aphis gossypii (Glover), and 53% mortality of the cotton leafhopper (Jassids) Amrasca bigutulla bigutulla (Ishida), while opposite results were observed in case of Bt at the highest concentration (1 × 108 cfu ml−1) used. In vivo results revealed that Ts caused a significant population reduction of the aphid (87%) than the jassid (72%), 7 days of post-treatment, at the highest concentration, while non-significant results were observed at the lowest concentrations. A. gossypii was significantly found more susceptible to the mixture of Trichoderma + Bt than the jassid (62%) and brinjal shoot and fruit borer (65%) even after 48 h of treatment application. The combined application (Trichoderma + Bt) showed maximum population reduction of jassid (88%), aphid (95%), and BSFB (96%), respectively, 7 days post-applications. The positive correlation among time and concentration was observed. The result may imply that consortium of these microbial organisms could be effective and can be incorporated in IPM programs for effective control of sucking and chewing insect pests of brinjal.

The hybrid seeds of several important crops with supreme qualities, including yield, biotic and abiotic stress tolerance, have been cultivated from decades. Thus far, a major challenge with hybrid seed, it does not hold ability to produce plants with same qualities over subsequent generations. Apomixis exist naturally an asexual mode of reproduction in flowering plants via avoiding meiosis and ultimately leads to seed production. Apomixis possess potential to preserve hybrid vigor for multiple generations for economically important plant genotypes. The evolution and genetics of asexual seed production is unclear and need much more efforts to find its genetic architecture. To fix hybrid vigor synthetic apomixis has been suggested an alternative. The development of MiMe (Mitosis instead of Meiosis) genotypes are utilized further for clonal gametes production. However, the identification and parental origin of genes responsible for synthetic apomixis are less known and need further understanding. Genome modifications utilizing genome editing technologies (GETs) like clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (cas9) a reverse genetics tool has paved way to utilize emerging technologies in plant molecular biology. From the last decade, several genes in important crops have been successfully edited. The vast availability of GETs has made the functional genomics studies easy to conduct in crops important for food security. The disruption of expression of genes specific to egg cell MATRILINEAL (MTL) or BABY BOOM1 (BBM1) through CRISPR/Cas genome editing system can promote haploid plants. The establishment of synthetic apomixis by engineering MiMe genotype by genome editing BBM1 expression or disruption of MTL leads toward clonal seed production. In present review, we discussed the current development in plants by utilizing CRISPR/Cas9 technology and its possibility of promoting apomixis in crops to preserve hybrid vigour. In addition to this, genetics, evolution, epigenetic modifications and strategy for MiMe genotype development has been discussed in detail.

The Nipa palm hispid, Octodonta nipae (Maulik) is an important invasive pest of palm trees particularly in Southern China. How this beetle interacts with invading pathogens via its immune system remains to be dissected. Steinernema carpocapsae is a pathogenic nematode that attacks a number of insects of economic importance. The present study systematically investigates the cellular immune responses of O. nipae against S. carpocapsae infection using combined immunological, biochemical and transcriptomics approaches. Our data reveal that S. carpocapsae efficiently resists being encapsulated and melanized within the host's hemolymph and most of the nematodes were observed moving freely in the hemolymph even at 24 h post incubation. Consistently, isolated cuticles from the parasite also withstand encapsulation by the O. nipae hemocytes at all-time points. However, significant encapsulation and melanization of the isolated cuticles were recorded following heat treatment of the cuticles. The host's phenoloxidase activity was found to be slightly suppressed due to S. carpocapsae infection. Furthermore, the expression levels of some antimicrobial peptide (AMP) genes were significantly up-regulated in the S. carpocapsae-challenged O. nipae. Taken together, our data suggest that S. carpocapsae modulates and surpasses the O. nipae immune responses and hence can serve as an excellent biological control agent of the pest.

Wolbachia pipientis is a diverse, ubiquitous and most prevalent intracellular bacterial group of alpha-Proteobacteria that is concerned with many biological processes in arthropods. The coconut hispine beetle (CHB), Brontispa longissima (Gestro) is an economically important pest of palm cultivation worldwide. In the present study, we comprehensively surveyed the Wolbachia-infection prevalence and mitochondrial DNA (mtDNA) polymorphism in CHB from five different geographical locations, including China’s Mainland and Taiwan, Vietnam, Thailand, Malaysia and Indonesia. A total of 540 sequences were screened in this study through three different genes, i.e., cytochrome oxidase subunit I (COI), Wolbachia outer surface protein (wsp) and multilocus sequencing type (MLST) genes. The COI genetic divergence ranges from 0.08% to 0.67%, and likewise, a significant genetic diversity (π = 0.00082; P = 0.049) was noted within and between all analyzed samples. In the meantime, ten different haplotypes (H) were characterized (haplotype diversity = 0.4379) from 21 different locations, and among them, H6 (46 individuals) have shown a maximum number of population clusters than others. Subsequently, Wolbachia-prevalence results indicated that all tested specimens of CHB were found positive (100%), which suggested that CHB was naturally infected with Wolbachia. Wolbachia sequence results (wsp gene) revealed a high level of nucleotide diversity (π = 0.00047) under Tajima’s D test (P = 0.049). Meanwhile, the same trend of nucleotide diversity (π = 0.00041) was observed in Wolbachia concatenated MLST locus. Furthermore, phylogenetic analysis (wsp and concatenated MLST genes) revealed that all collected samples of CHB attributed to same Wolbachia B-supergroup. Our results strongly suggest that Wolbachia bacteria and mtDNA were highly concordant with each other and Wolbachia can affect the genetic structure and diversity within the CHB populations.

American bollworm (ABW), Helicoverpa armigera (Hubner), is considered as a major pest of cotton, Gossypium hirsutum, all over the globe. Due to its destructive feeding nature and continuous consumption of the same chemicals, it devolved resistant against many insecticides. Therefore, a combined application of bio- and synthetic-pesticide need to evaluate against this pest. The entomopathogenic viruses like nuclear polyhedrosis virus (NPV), a member of baculoviruses, can be the potential candidates for better control against ABW. The present study was conducted to assess the comparative efficacy of NPV and Spinosad 240SC (with the concentration of 250 mL · ha‾ˡ) against ABW in the controlled environment. The ABW was treated with different concentrations of NPV and Spinosad separately and in a combination of NPV with 0.1% Spinosad. The results revealed that highest concentrations showed highest mortality (95%) followed by 95%, 92%, 84%, 82% and 78% mortality at 1 × 109, 1 × 108, 1 × 107, 1 × 106 and 1 × 105 POBs, respectively. Spinosad when mixed in diet give 100% mortality at 0.8% followed by 50.87%, 42.10%, 29.82%, 26.31% and 22.80% mortality at 0.4%, 0.2%, 0.1%, 0.5% and 0.025% respectively. The results of this study revealed that microbial control of ABW through NPV is an effective tool. The repeated use of synthetic pesticides caused the resurgence of many insect pests, and this study results would provide useful insight to build a framework for future investigations for the management of many major insect pests.

Studies on the insect pollinators diversity and their relative abundance in Eruca sativa Mill. (Arugula) and Brassica rapa L. (field mustard) was carried out during spring season from February to April consecutively during all the three years of 2016–18. Insect pollinators observed belonged to four orders i.e. Hymenoptera, Diptera, Lepidoptera, and Coleoptera. A total of 20 major species of insect pollinators were recorded. The highest abundance of pollinator species belonged to Hymenoptera. The most prominent insect pollinator species were Apis mellifera followed by other three honey bee species of A. cerana, A. florea, and A. dorsata respectively. Some species of solitary bees were also recorded. From Diptera, four species of syrphid fly and one species from Muscidae family were also recorded. Insect pollinators recorded from order Lepidoptera were Pieris brassicae, Vanessa cardui, and Papilio demoleus. Lady bird beetle Coccinella septempunctata was recorded from Coleoptera order as occasional visitor. It was noticed that E. sativa attracted more insect pollinators than B. rapa which may be attributed to different amount and chemical properties of nectar, with number of pollen grains, and flower canopy of both crops. Further studies are needed to confirm the reasons for higher pollinator visitation to E. sativa than B. rapa through chemical analysis of nectar, amount of pollens, flower physiology and phenology of both crops. Keywords: Insect pollinators, Diversity, Hymenoptera, Diptera, Coleoptera, Lepidoptera, Brassicaceae, Cruciferous crops

The whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is a cryptic species complex distributed worldwide. In Pakistan, B. tabaci poses a serious threat to agriculture production. To understand its diversity in Pakistan, a large-scale sampling was conducted from various locations of all four provinces of the country and Mitochondrial cytochrome oxidase I (mtCOI) gene sequencing was used to determine the whiteflies genetically. The study revealed the presence of five different cryptic species in Pakistan namely Asia II-1, Asia II-5, Asia II-7, Asia II-8 and MEAM-1, respectively. Among them, Asia II-1, which was previously reported from a few areas in the country, had been found now to be prevalent all over the country covering 88.7% of all the sequenced samples. Based on the mtCOI sequences and genetic distance analyses, the diversity of Asia II-1 was much greater than all other cryptic species, which exist only in small patches.

The intracellular bacterium Wolbachia pipientis is widespread in arthropods. Recently, possibilities of novel Wolbachia-mediated hosts, their distribution, and natural rate have been anticipated, and the coconut leaf beetle Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), which has garnered attention as a serious pest of palms, was subjected to this interrogation. By adopting Wolbachia surface protein (wsp) and multilocus sequence type (MLST) genotypic systems, we determined the Wolbachia infection density within host developmental stages, body parts, and tissues, and the results revealed that all the tested samples of B. longissima were infected with the same Wolbachia strain (wLog), suggesting complete vertical transmission. The MLST profile elucidated two new alleles (ftsZ-234 and coxA-266) that define a new sequence type (ST-483), which indicates the particular genotypic association of B. longissima and Wolbachia. The quantitative real-time PCR analysis revealed a higher infection density in the eggs and adult stage, followed by the abdomen and reproductive tissues, respectively. However, no significant differences were observed in the infection density between sexes. Moreover, the wsp and concatenated MLST alignment analysis of this study with other known Wolbachia-mediated arthropods revealed similar clustering with distinct monophyletic supergroup B. This is the first comprehensive report on the prevalence, infection dynamics, and phylogeny of the Wolbachia endosymbiont in B. longissima, which demonstrated that Wolbachia is ubiquitous across all developmental stages and distributed in the entire body of B. longissima. Understanding the Wolbachia infection dynamics would provide useful insight to build a framework for future investigations, understand its impacts on host physiology, and exploit it as a potential biocontrol agent.

The humoral immune responses of the nipa palm hispid beetle Octodonta nipae involves the inducible expression of the genes coding for antimicrobial peptides (AMPs) which are mediated by immune deficiency signaling pathways. In insects, the nuclear factor-κB (NF-κB) transcription factor, Relish, has been shown to regulate AMP gene expressions upon microbial infections. Here, we dissect the expression patterns of some AMPs in O. nipae during infections by entomopathogenic nematodes (EPNs) and their symbionts, before and after Relish knock down. Our results indicate that, prior to gene silencing, the AMPs attacin C1, attacin C2, and defensin 2B were especially expressed to great extents in the insects challenged with the nematodes Steinernema carpocapsae and Heterorhabditis bacteriophora as well as with their respective symbionts Xenorhabdus nematophila and Photorhabdus luminescens. The study also established the partial sequence of OnRelish/NF-κB p110 subunit in O. nipae, with an open reading frame coding for a protein with 102 amino acid residues. A typical Death domain-containing protein was detected (as seen in Drosophila) at the C-terminus of the protein. Phylogenetic analysis revealed that in O. nipae, Relish is clustered with registered Relish/NF-κB p110 proteins from other species of insect especially Leptinotarsa decemlineata from the same order Coleoptera. Injection of OnRelish dsRNA remarkably brought down the expression of OnRelish and also reduced the magnitude of transcription of attacin C1 and defensin 2B upon S. carpocapsae and H. bacteriophora and their symbionts infections. Altogether, our data unveil the expression pattern of OnRelish as well as that of some AMP genes it influences during immune responses of O. nipae against EPNs and their symbionts.

In order to interpret the molecular mechanisms that modulating the organism variations and selection signatures to drive adaptive evolutionary changes are indispensable goals in the new evolutionary ecological genetics. Here, we identified the gene locus associated to royal jelly production through whole-genome sequencing of the DNA from eight populations of honeybees. The analysis of the samples was composed of 120 individuals and each pointed extremely opposite trait values for a given phenotype. We identified functional single nucleotide polymorphisms (SNPs) candidate that might be essential in regulating the phenotypic traits of honeybee populations. Moreover, selection signatures were investigated using pooling sequencing of eight distinct honeybee populations, and the results provided the evidence of signatures of recent selection among populations under different selection objectives. Furthermore, gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that selected genes were potentially involved in several biological processes and molecular functioning, which could directly or indirectly influence the production of royal jelly. Our findings can be used to understand the genomic signatures, as well as implicate a profound glance on genomic regions that control the production trait of royal jelly in honey bees.

Plants under natural environment facing various pathogens, tend to produce defense to maintain their fitness and minimize pathogenic damage. Plant-pathogens interaction is gaining more importance by researches as, their means of the fight are primary metabolites. The ultimate result of either means of defense is pathogenesis or resistance. Plant defense mechanisms can be grouped either into inducible and constitutive defense or chemical, structural and morphological defense. Majority of defense mechanisms have a passive role, i.e. only defensive against pathogens, but a few are very active. Plant primary metabolites are catching interest in their immunity role. Deep information of molecular mechanisms involved during the plant-pathogen system is need of the day for future disease control. This review will highlight the role of primary metabolites and their mechanism of action in plant defense.

Induction of stress-related genes is primarily reckoned as a major step in plant responses to any stress. The plant genomes possess multiple types of transcription factors (TFs) belonging to diverse families and many of them are unique to plants. The TFs are regulators of transcriptional reprogramming linked with stress responses in plants. TF gene(s) frequently retorts to numerous stresses and then their respective polypeptides may contribute in the positive or negative regulation of apparently dissimilar processes. Many plant TFs are induced by biotrophic as well as necrotrophic pathogens. Functional characterization of TFs provides solid evidence of their role in the regulation of plant defense responses. This advocates the fact that TF does not work alone, but the immune response to pathogens mediated by it, is strongly linked to enhancing the effect of TF on the transcription of defense associated genes. The plant-plant difference in transcriptional responses of TF and Defense associated genes highlights that a TF may regulate defense positively or negatively. This diversity in responses is proof of functional diversity among TFs. Their functions in plant immunity and abiotic stress tolerance in plants have been characterized but still need investigations in Capsicum annuum. This article focuses upon recent progress in our understanding of the role of the TFs involved in Capsicum defense against Microbial pathogenesis especially against bacterial pathogens. We have dissected collateral interactions between TFs, proteins and nuclear receptors working for Capsicum defense. Besides, we have also reviewed and discussed the topical concepts of transcriptional intervention for controlling the activity of TFs.

The present study was conducted to highlight the existing level of organochlorine-pesticides (OCPs) from human milk (n = 45) and blood serum (n = 40) of female workers who pick cotton in Khanewal District, southern Punjab, Pakistan. Source apportionment, congener-specific analysis, and risk surveillance of OCPs are reported from human milk and blood samples. Levels of OCPs in milk and blood serum samples ranged from 15.7 ppb to 538.3 ppb and from 16.4 ppb to 747.1 ppb, respectively, and were lower than previously published reports from other regions of the globe. Congener-specific analysis revealed that DDTs were predominant, followed by hexachlorocyclohexane, chlordane, and hexachlorobenzene. Calculated results for source apportionment analysis suggested that contamination load was a new input of DDTs as well as the historic use of lindane in the study area. Levels of OCPs in milk and blood serum were significantly (p

The endo-symbiotic relationship between microorganism and eukaryotes are very common and has been broadly investigated from all insect species. The genus Wolbachia are obligatory intracellular bacteria that induce evolutionary alterations and have been frequently reported in egg cytoplasm’s of various invertebrates including insects, spiders, mites, scorpions, crustaceans and nematodes, which exert a profound impact on host biology and behavior through a number of phenotypic alternations. Due to its ubiquitous phenotypic behavior, Wolbachia becomes a novel and promising natural micro-biocontrol agent to induce cytoplasmic incompatibility, killing of male embryos, parthenogenesis, and feminization. Autonomous transgenic capability, survival on several host species and the ability to modify the host reproductive systems makes it the most prominent and focusing scientific genomic research from last few decades. The aim of this review is to summarize the significance and recent discoveries of Wolbachia related to various arthropods which lead positive directions to be exploited in future for integrated pest management (IPM) strategies. Further, this review also discussed the biology, phylogeny, distributions, classifications and types of Wolbachia infections on insect orders and coleopterans in particular.

Salinity is a major factor limiting irrigated agriculture in arid regions. Vermi amendments can be used for improving the fertility of salt-affected soils. Current study was aimed to find out the response of different earthworm species to soil salinity and to check the effects of salinity and different vermi amendments on growth of Sorghum bicolor under salt stress. Eight earthworm species were subjected to different salinity levels for 4 weeks. Various vermi amendments and salinity treatments were provided in a factorial combination to S. bicolor plants to see their effect on growth and biomass parameters. L. mauritii, E. incommodus and P. posthuma were found to be the most salt-tolerant species showing good survival and growth till soil ECe value of 10.48 mS cm−1. Results showed that salinity significantly decreased plant growth that was enhanced by the application of different vermi amendments. Maximum growth of S. bicolor was recorded when vermicompost and vermiwash were used together under bo...

The study was carried out in eight provinces of China, with 16 different localities. A total of 150 adult damselflies were collected for the study from the area. Mitochondrial (COI) was the target gene to sequence from all 51 representative samples of the entire collection. From the resulted sequences, ten species belonging to seven genera and four families were identified. The families are Calopterygidae, Coenagrinidae, Chlorocyphidae and Devadattiade. After alignment through BioEdit v6, MEGA7 was used for phylogenetic tree construction as well as the calculation of genetic divergence, whereas genetic diversity was calculated by DnaSP v5. All species from the respective families had clustered together within the groups, but divided into sub-groups. Maximum genetic divergence (9.31%) was found in the genus Rhinocypha, followed by Hetaerina with (8.23%), while, minimum divergence was observed for the genus Ceriagrion (0.32%) followed by Ischnura (0.48%). However, significant genetic diversity was found for all sequences at 346 mutations confirmed by two tests, Tajima's D and Fu's Fs. Maximum genetic diversity among genera was also observed, the genus Hetaerina has maximum genetic diversity (181 mutations) followed by Rhinocypha (68 mutations) while minimum genetic diversity was observed for the genus Ceriagrion followed by Ischnura. The presented results showed a higher diversity of damselflies in the south China regions.

To determine population genomic structure through high-throughput sequencing techniques has revolutionized research on non-model organisms. The coconut leaf beetle, Brontispa longissima (Gestro), is a widely distributed pest in Southern China. Here, we used restriction site-associated DNA (RAD) genotyping to assess the invasion pathway by detecting and estimating the degree of genetic differentiation among 51 B. longissima accessions collected from Southern China. A total of 10,127 SNPs were obtained, the screened single nucleotide polymorphism (SNP) information was used to construct the phylogenetic tree, FST analysis, principal component analysis, and population structure analysis. Genetic structure analysis was used to infer the population structure, the result showed that all accessions were divided into Hainan population and non-Hainan population. The Hainan population remained stable, only the Sansha population differentiated, and the non-Hainan populations have gradually differentiated into smaller sub-populations. We concluded that there are two sources of invasion of B. longissima into mainland China: Taiwan and Hainan. With the increase of the invasion time, the Hainan population was relatively stable, and the Taiwan population was differentiated into three sub-populations. Based on the unrooted phylogenetic tree, we infer that Taiwan and Hainan are the two invasive base points. The Taiwan population invaded Fujian, Guangdong, and Guangxi, while the Hainan population invaded Yunnan and Sansha. Our results provide strong evidence for the utility of RAD sequencing (RAD-seq) in population genetics studies, and our generated SNP resource could provide a valuable tool for population genomics studies of B. longissima in the future.

Microbial consortia accompanied to all eukaryotes can be inherited from ancestors, environment, and/or from various food source. Gut microbiota study is an emerging discipline of biological sciences that expands our understanding of the ecological and functional dynamics of gut environments. Microorganisms associated with honey bees play an important role in food digestion, colony performance, immunity, pollination, antagonistic effect against different pathogens, amelioration of food and many more. Although, many repots about honey bee gut microbiota are well documented, microbiome with other key components of honey bees such as larvae, adults, their food (pollen, beebread, and honey), honey combs, and floral nectar are poorly understood. Mutual interactions and extent of the roles of microbial communities associated with honey bees are still unclear and demand for more research on the nutritional physiology and health benefits of this ecologically and economically important group. Here in this study, we highlighted all the honey bee microbiome that harbored from different life stages and other relevant components. The anatomical parts of honey bee (larvae, adults), food source (pollen, beebread, and honey), honey combs, and floral nectar were highly flourished by numerous microorganisms like bacteria (Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Actinobacteria, Actinomycetes, Bacilli, Bacteroidetes, Cocci, Clostridia, Coliforms, Firmicutes, Flavobacteriia, Mollicutes) and fungi (Dothideomycetes, Eurotiomycetes, Mucormycotina, Saccharomycetes, Zygomycetes, Yeasts, Molds). Some distinctive microbial communities of a taxonomically constrained species have coevolved with social bees. This contribution is to enhance the understanding of honey bee gut microbiota, to accelerate bee microbiota and microbiome research in general and to aid design of future experiments in this growing field.

Adult dragonflies (Anisoptera) were collected from different localities of South China covering eight provinces. Representative sequences were sixty-one, including 16 species, 11 genera and three families (Aeshnidae, Gomphidae and Libellulidae), under cytochrome oxidase subunit I (COI) gene. After alignment of sequences by BioEdit v6, genetic interaction and divergence were computed by MEGA 7 whereas all the indices of genetic diversity were calculated by DnaSP v5 software. Phylogenetic trees were constructed through Neighbor-Joining method under Jukes-Cantor model, and all species of respective families were assembled with each other into individual groups. Maximum divergence was observed by Trithemis genus (18.69%), followed by Orthetrum genus (18.16%), whereas a minimum value of divergence was noted for Pantala genus (0.31%). On the other hand, maximum genetic diversity was recorded for Orthetrum genus up to 142 mutations, followed by Trithemis genus (126 mutations), while the minimum value (two mutations) was observed for Pantala genus. Genetic diversity for overall and Libellulidae family sequences was much higher, up to 404 mutations and 344 mutations, respectively. Current results suggest a high diversity of odonates in the South China region and results are valuable in gaining a total obligation of the diversity of Asian odonates and conservation measures of this insect group.

The Asian citrus psyllid (ACP), Diaphorina citri Kuwayama (Hemiptera: Psyllidae) is a devastating pest of Citrus spp. The aim of present study was to investigate the development and mortality of ACP on citrus (Citrus sinensis) (healthy and Huanglongbing- (HLB) diseased) and jasmine (Murraya paniculata) plants at various temperatures. Two new Isaria strains were collected from citrus orchards of Fuzhou (China), and HLB-diseased plants were verified by running PCR for 16S gene of Candidatus Liberibacter asiaticus (CLas). Development observations were recorded for egg, nymph and adult stages on all plants and three different temperatures (20, 25 and 30 °C) whereas mortality observations were recorded for the nymph (fifth instar) and adults on all plants at 25 °C. Field collected Isaria strains were belonged to previously reported Chinese strains under Maximum Parsimony (MP) and Maximum Likelihood methods, as well as, CLas isolates were belonged to previously reported Chinese isolates under MP and Neighbor-Joining methods. The fastest development and mortality was observed on HLB-diseased plants whereas longest time was taken by development and mortality completion on jasmine plants at all temperatures. The fastest developmental times of egg, nymph (first to fourth and fifth instar) and adult stages were ranged from 3.02 to 3.72 d, to 7.63–9.3 d, 5.35–5.65 d and 24.46–28.47 d on HLB-diseased plants at 30-20 °C, respectively. On the other hand, I. javanica caused the fastest mortality of nymphs and adults (32.21 ± 4.47% and 19.33 ± 4.51%) on HLB-diseased plants with the concentration of 1 × 108 conidia.mL−1 after 3 d and 7 d, respectively. It is concluded that there is a need for extensive molecular work to understand the extra-development and mortality of ACP on diseased plants, because, CLas bacterium can be supportive to uptake more sap from plant phloem.

To elucidate taxonomic eminence of identical pest species is essential for many ecological and conservation studies. Without proficient skills, accurate molecular identification and characterization are laborious and time-consuming. The coconut leaf beetle, Brontispa longissima (Gestro) (Coleoptera: Chrysomelidae), is biologically and morphologically identical to hispid leaf beetle, Octodonta nipae (Maulik) (Coleoptera: Chrysomelidae), and is known as the most harming nuisances of palm cultivation worldwide. The present examination was to establish Wolbachia genotyping analysis along with host cytochrome oxidase subunit I (COI) gene for accurate identification between these individuals of the same family (Chrysomelidae). Here, we have cloned and sequenced a gene coding Wolbachia surface protein (wsp) and COI gene regions amplified from both species by polymerase chain reaction. The nucleotide sequences were directly determined (≈600 bp for wsp and ≈804 bp for COI) and aligned using the multiple alignment algorithms in the ESPript3 package and the MEGA5 program. Comparative sequence analysis indicated that the representative of wsp and COI sequences from these two beetles were highly variable. To ensure this bacterial variation, multilocus sequence typing (MLST) of bacterial genes was conducted, and the results vindicated the same trend of variations. Furthermore, the phylogenetic analysis also indicates that B. longissima and O. nipae being the two different species harbors two distinct Wolbachia Hertig and Burt (Rickettsiales: Anaplamataceae) supergroups B and A, respectively. The present outcomes quickly discriminate between these two species. Considering its simplicity and cost-effectiveness, it can be used as a diagnostic tool for discriminating such invasive species particularly B. longissima and O. nipae which has overlapping morphologic characters.

The hispid leaf beetle, Octodonta nipae (Maulik), (Coleoptera: Chrysomelidae), is a devastating pest of palm cultivation worldwide. Endosymbiotic bacteria in the genus Wolbachia are arguably one of the most abundant bacterial group associated with arthropods. Owing to its critical effects on host reproduction, Wolbachia has garnered much attention as a prospective future tool for insect pest management. However, their association, infection dynamics, and functionality remain unknown in this insect pest. Here, we diagnosis for the first time, the infection prevalence, and occurrence of Wolbachia in O. nipae. Experimental evidence by the exploration of wsp gene vindicate that O. nipae is naturally infected with bacterial symbiont of genus Wolbachia, showing a complete maternal inheritance with shared a common Wolbachia strain (wNip). Moreover, MLST (gatB, fbpA, coxA, ftsZ, and hcpA) analysis enabled the detections of new sequence type (ST-484), suggesting a particular genotypic association of O. nipae and Wolbachia. Subsequently, quantitative real-time PCR (qPCR) assay demonstrated variable infection density across different life stages (eggs, larvae, pupae and adult male and female), body parts (head, thorax, abdomen), and tissues (ovaries, testes, and guts). Infection density was higher in egg and female adult stage, as well as abdomen and reproductive tissues as compared to other samples. Interestingly, Wolbachia harbored dominantly in a female than the male adult, while, no significant differences were observed between male and female body parts and tissues. Phylogeny of Wolbachia infection associated with O. nipae rectified from all tested life stages were unique and fall within the same monophyletic supergroup-A of Wolbachia clades. The infection density of symbiont is among the valuable tool to understand their biological influence on hosts, and this latest discovery would facilitate the future investigations to understand the host-symbiont complications and its prospective role as a microbiological agent to reduce pest populations.

A total of 300 dragonflies (Odonata) were collected from six different localities of China and Pakistan. Sixty seven representative samples were selected to sequence their mitochondrial cytochrome oxidase subunit I (COI). An examination of the resultant sequences identified 21 different dragonfly species, belonging to 15 distinct genera, two families, Libellulidae and Gomphidae. Sequence alignment was executed using Clustal-W in BioEdit v6. The phylogenetic tree was constructed through Neighbor-joining method by using Jukes-Cantor model, and genetic divergence was calculated via Kimura 2-parameter using MEGA7, while Genetic diversity was calculated by DnaSP v5. The maximum genetic divergence was observed for Crocothemis servilia, at 20.49%, followed by Libellulidae sp. with 22.30% while minimum divergence (0.82%) was observed for Melligomphus ardens. Likewise, a significant genetic diversity was observed for all species. However, Crocothemis servilia species presented maximum value (176 mutations) followed by Libellulidae spp. (150 mutations), whereas minimum value (3 mutations) was observed by Orthetrum testaceum. Interestingly, the diversity of C. servilia, all of which are collected from a single location of China, is much higher than those from Pakistan, which were collected from 5 different places with a spatial distance exceeding 500 Kms. Our results are useful in gaining a full appreciation of the global diversity of dragonflies and the development of conservation measures of this insect.

ObjectiveTo appraise the awareness and knowledge levels of midwives and nurses concerning early screening for Zika virus (ZIKV) infection among pregnant women attending health care facilities in Bahrain.MethodsThis was a cross-sectional, purposely chosen study of Bahraini and expatriate midwives, nurses, and supervisors employed in gynecology/obstetrics and labor wards of Salmanya hospital, a maternity hospital, 4 private hospitals, and health centers in Bahrain. The chosen individuals were invited to participate in a survey on awareness and knowledge of early screening for ZIKV infection.ResultsOf 266 midwives and nurses employed in the study sites, 170 (64%) consented to participate in the study. Of those who agreed to participate, 76 were midwives and 94 were nurses. Admittedly, 39% of midwives and nurses were unaware of ZIKV infection. The grand mean knowledge score in the study was 39%. Expatriate midwives and nurses scored better than did Bahrainis (PConclusionsThe awareness and knowledge scores of midwives and nurses concerning ZIKV infection were inadequate, which supported our hypothesis. By harnessing modern technology and support systems, lifelong learning can be used as a means to enhance preparedness for public health crises such as ZIKV. (Disaster Med Public Health Preparedness. 2018;12:7–13)

Surface Plasmon Resonance (SPR) biosensors have emerged as a promising technique for bioanalysis as well as microbial detection and identification. Real time, sensitive, cost effective, and label free detection of biomolecules from complex samples is required for early and accurate diagnosis of infectious diseases. Like many other types of optical techniques, SPR biosensors may also be successfully utilized for microbial detection for accurate, point of care, and rapid results. In the present study, we have utilized a commercially available automated SPR biosensor of BI company to study the microbial detection form water samples spiked with different concentration of Staphylococcus aureus bacterial cells. The gold thin film sensor surface was functionalized to react with proteins such as protein G, which was used for directec immobilization of monoclonal antibodies against Staphylococcus aureus. The results of our work reveal that this immunosensor can be used to detect very small number of bacterial cells with higher sensitivity and specificity. In our case 103 cells/ ml of water have been successfully detected. Therefore, it may be concluded that this technique has a strong potential to be used in microbial detection and identification.

We investigated nesting of Barn Owls (Tyto alba) in central Punjab, Pakistan, from 1999–2002. We also documented diet of Barn Owls in southern Punjab and southeastern Baluchistan, Pakistan. Of 40 nests found in central Punjab, 33 were located in tree hollows, four in buildings, and three in nest boxes. Egg-laying was observed primarily between the second week of August and October (N = 23), with a smaller number of nests started May–July (N = 5). Fledging of the autumn nests was completed by early January, and all breeding efforts observed in May–July were unsuccessful. During the August–January breeding period, clutch size averaged 5.83 eggs (SE = 0.47; range = 1–12; N = 23) and brood size 4.15 nestlings (SE = 0.28; range = 2–7; N = 20). Shrews (59%), birds (19%), rats and mice (13%), and insects (7%) were the main prey in southern Punjab, whereas rats and mice (93%), shrews (4%), and amphibians and reptiles (3%) were the main prey in southeastern Baluchistan.