Evaluation of Insecticidal Potentials of Five Plant Extracts against the Stored Grain Pest, Callosobruchus maculatus (Coleoptera: Bruchidae).

Plant based insecticides are considered among the most economic and ecofriendly chemicals for the protection of plants and stored grains. The cowpea weevil (Callosbruchus maculatus) causes more than 90% damage to sored grains in three to six months. The current study investigates insecticidal potentials of five selected botanicals: Melia azedarach, Nicotiana rustica, Azadirachta indica, Nicotiana tabacum and Thuja orientalis. They are explored at six different concentrations (0.5, 1.0, 1.5, 2.0, 2.5 and 3.0%) against C. maculatus and compared to effects of distilled water which is used as a control. Toxicities of 3%(V/V) extracts of N. tabacum, N. rustica, A. indica and T. orientalis against C. maculatus were 100%, 86.11%, 80.56% and 72.22%, respectively. Maximum mortality was caused by N. tabacum and N. rustica (100%), followed by A. indica (82%), whereas minimum mortality was observed in T. orientalis (64%) at 2.5%. Several phytochemicals, alkaloids, saponins, diterphenes, phytosterol, flavonoids and phenols were identified in N. tabacum and N. rustica, while few were present in A. indica. Phytosterol was present in greatest abundance. Saponins were only detected in aqueous extracts of N. rustica and N. tabacum. Taken together, these results indicate the utility of N. tabacum, N. rustica and A. indica as potential botanicals to control pest beetle and cowpea weevil.

A Novel Protein Elicitor (PELL1) Extracted from Lecanicillium lecanii Induced Resistance against Bemisia tabaci (Hemiptera: Aleyrodidae) in Gossypium hirsutum L.

Protein elicitors play a key role in signaling or displaying plant defense mechanism and emerging as vital tools for biocontrol of insects. This study was aimed at the characterization of the novel protein elicitor isolated from entomopathogenic fungi Lecanicillium lecanii (V3) strain and its activity against whitefly, Bemisia tabaci, in cotton (Gossypium hirsutum L.). The sequence of purified elicitor protein showed 100% similarity with hypothetical protein LEL_00878 (Cordyceps confragosa RCEF 1005) (GenBank accession no. OAA81333.1). This novel protein elicitor has 253 amino acid residues and 762 bp with a molecular mass of 29 kDa. Their combatant protein was expressed in Escherichia coli using pET-28a (+) plasmid. Bioassay was revealed to quantify the impact of numerous concentrations of protein (i.e., 58.32, 41.22, and 35.41 µg/ml) on the fecundity rate of B tabaci on cotton plants. Bioassay results exhibited a significant effect (P ≤ 0.001) of all the concentrations of protein on the fecundity rate of B. tabaci. In addition, the gene expression analysis found a significant upregulation of the major genes associated with salicylic acid (SA) and jasmonic acid (JA) defense pathways in elicitor protein-treated plants. Our results showed that the potential application of novel protein elicitor derived from Lecanicillium lecanii will be used as future biointensive controlling approaches against whitefly, Bemisia tabaci.

Ameliorative effect of L-carnitine on lambda-cyhalothrin-induced anatomical and reproductive aberrations in albino mice.

The objective of this research was to look at how the pesticide lambda-cyhalothrin (LCT) affected the liver, kidney, testis, and ovary of albino mice; and on morphological and skeletal features of the newborn of treated females. The study also aimed to test the ameliorative effects of L-carnitine (LC) against (LCT). Five sets of mice were created, Group 1 acted as the control, while Group 2 received a high dose of LCT, Group 3 received a high dose of LCT + LC, Group 4 received a low dose of LCT that was a residue of in khat (Qat), and Group 5 received a low dose of LCT + LC. The findings revealed that the treated groups' body weights were reduced significantly, whereas the absolute and relative weights of the liver in all groups were statistically decreased insignificantly. There were histopathological changes in the tissues in groups 2 and 4. While the tissues of the ovary and testis showed recovery in groups 3 and 5. When compared to the control group, the values of the seminiferous tubules parameters were statistically significant in the 3 and 5 groups. The newborn had a high dose of pesticides and showed some malformations in the skeleton. However, in group 3 the skeletal malformation was minimized and in-group 5 the skeleton malformations had completely disappeared. It could be concluded that LCT is highly harmful to mouse tissues and caused neonatal malformations, whereas LC has a marked protective effect against LCT.

Correction: Path analysis based on genetic association of yield components and insects pest in upland cotton varieties.

[This corrects the article DOI: 10.1371/journal.pone.0260971.].

Effect of temperature & humdity on population dynamics of insects' pest complex of cotton crop.

The current study was directed to investigate the effect of temperature and humidity on insect pest complex of cotton. This study was carried out on the farmer field of a farmer of MouzaMehraywala tehsil and district Rajanpur southern part of the Punjab, Pakistan (29.1044° N, 70.3301° E) in the month of May, 2019 to November 2019. The climatic conditions of the study site were; a Rajanpur lie on 96m above sea level Rajanpur has a desert climate. During the year, there is virtually no rainfall. The average temperature in Rajanpur is 26.0°C | 78.8°F. Precipitation here is about 205 mm / 8.1 inch per year. Our results indicate that's the correlation of temperature (minimum and maximum), humidity (minimum and maximum), rainfall and weather condition (clear or cloud), it is clear that except temperature there is a positive correlation of jassid population. While among the relation of jassid with different factor there is very weak relation with jassid population regarding temperature (-0.001), the relation of humidity, rainfall and sky condition with jassid population is positive and moderate (0.520, 0.668 & 0.575 respectively), while the relation of jassid population among these factor is significant except temperature. The results indicate that's the correlation of temperature, humidity, rainfall and weather condition, it is clear that there is positive correlation of thrips population. While among the relation of thrips with different factor there is very weak relation of thrips population with temperature (.103), the relation of humidity, rainfall and sky condition with thrips population is positive and moderate (.515.751& .577 respectively), while the relation thrips population among these factor is significant except temperature. The results indicate that's correlation of temperature, humidity, rainfall and weather condition, it is clear that there is positive correlation of Whitefly population. While among the relation of Whitefly with different factors there is very moderate relation of Whitefly population with temperature (.076), the relation of humidity, rainfall and sky condition with Whitefly population is negative and moderate (-.051.-.368 & -.559 respectively), while the relation Whitefly population among these factor is significant except temperature.

Microbiological investigation study for Apis mellifera yemenitica and Apis mellifera carnica bee venoms on selected bacterial strains.

Bees are one of the ancient and the most social insects worldwide. They are of great economic and medical importance. Bee venom (BV) has many therapeutic effects and has been used since ancient times for the treatment of many diseases. The present study aimed to evaluate and compare the antibacterial effect of BV from two different bee subspecies Apis mellifera yemenitica (A. m. yemenitica) (indigenous strain) and Apis mellifera carnica (A. m. carnica) (carniolan strain) against selected Gram-positive and Gram-negative bacteria. Experimentally, venoms were extracted using an electrical venom collector from honey bee colonies of the subspecies, A. m. yemenitica and A. m. carnica, in Hail, Saudi Arabia. Each venom was tested against selected medically important Gram-negative strains, Salmonella Typhimurium, Pseudomonas aeruginosa, and Escherichia coli, while Staphylococcus aureus was selected as Gram-positive test organism. The minimum inhibitory concentration (MIC) method was used to compare the effect of BV from the two subspecies on the growth of the selected bacterial strains. Results showed that BV from both subspecies could equally inhibit the growth of Salmonella Typhimurium, Pseudomonas aeruginosa, and Escherichia coli at an MIC of 10 mg/ml. However, S. aureus was inhibited by an MIC of 5 and 10 mg/ml of BV from A. m. carnica and A. m. yemenitica, respectively. This suggested that the BV of the carnica subspecie was more inhibitory to this Gram-positive pathogen than its counterpart produced by the yemenitica subspecies. The present study shows that bee venom has a promising antibacterial effect.

Awareness and perception of malaria and dengue at school and college level in the district of Multan.

The purpose of this study is to examine the awareness and perception of malaria and dengue fever in Multan Punjab, Pakistan while taking into account the important role of government policies and other variables. The goal of this study is to examine the awareness of students in Multan, Pakistan on malaria and dengue. This study is based on a quantitative approach of secondary evidence from scientific journals and questionnaire surveys. It is also based on observational evidence gathered in Multan Punjab Pakistan, in a field study. The survey with school children, teachers and healthcare professionals were both formal and semi-structuralize. Studies have found that malaria and dengue mainly affect children's schooling through their absence, but can also induce brain loss and cognitive disability. In questionnaires, students were seen to have different understanding of the illness, but also to be able to serve as agents of health reform only through teachers. A sample size of 500 respondents has been selected from different colleges of district Multan Punjab, Pakistan. Correlation technique is used for the data analysis. According to our results it is concluded that the students at college level are aware of malaria and dengue diseases, but they are not capable of engaging and serving as agents for health reform. On the basis of results it is recommended that students must teach about epidemics diseases regarding how to handle these diseases.

Path analysis based on genetic association of yield components and insects pest in upland cotton varieties.

Gossypium hirsutum L. is also called upland cotton or Mexican cotton. It is the most widely cultivated species of cotton in the whole world. Globally, about 90% of all cotton production comes from the cultivars derived from this species. Some genetic parameters like monopodial branches per plant, sympodial branches per plant, sympodial branch length, bolls per plant, boll weight, sympo-boll distance, Ginning Out Turn%, staple length (rg = 0.9199**), and fiber strength along with seed cotton yield were evaluated for their potential utilization via selection in seed cotton yield improvement. Significant positive genetic correlations were estimated for monopodial branches per plant (rg = 0.9722**), sympodial branches per plant (rg = 0.7098**), sympodial branch length (rg = 0.617**), bolls per plant (rg = 0.8271**), boll weight (rg = 0.8065**), sympo-boll distance (rg = 0.6507**), Ginning Out Turn (GOT)% (rg = 0.7541**), staple length (rg = 0.9199**), and fiber strength (rg = 0.7534**) with seed cotton yield. A path analysis of all the yield traits under study revealed strong positive direct effects of monopodial branch length (1.1556), sympo-boll distance (0.8173) and staple length (0.7633), while plant height exerted a highly strong direct negative effect (-1.2096) on yield. It is concluded that a direct selection based on monopodial branch length and sympo-boll distance, and staple length is effective, whereas, monopodial branch length, and sympodial branch length are good selection indicators via bolls per plant for yield improvement in cotton.

Prevalence Rate and Molecular Characteristics of Oestrus ovis L. (Diptera, Oestridae) in Sheep and Goats from Riyadh, Saudi Arabia.

Heads of sheep (n = 600) and goats (n = 800) slaughtered at Al-Aziziah Abattoir in Riyadh, Saudi Arabia, were inspected for the presence of O. ovis larvae (L). Heads were split along the longitudinal axes, and larvae (L1, L2, and L3) were gathered. The infestation rate was significantly higher in goats (44.5%; 356/800) than that in sheep (22.3%; 134/600). Out of the 151 collected larvae from sheep, 0% were L1, 1.3% were L2, and 98.7% were L3. Out of the total of 468 larvae from goats, 0% were L1, 1.2% were L2, and 98.8% were L3. The infestation rate was significantly higher in males than that in females. Myiasis-causing larvae collected from Riyadh, Saudi Arabia, were authenticated as O. ovis, according to morphological characteristics. Polymerase chain reaction (PCR) amplification of a partial fragment (600 bp) of the mitochondrial cytochrome c oxidase subunit I (mtCOI) gene further confirmed the species. Phylogenetic analysis based on the partial mtCOI gene sequence demonstrated that 23 unique sequences showed high similarity based on nucleotide pairs of O. ovis accessions retrieved from GenBank.

Molecular identification of Campanulotes bidentatus Scopoli, 1763 (Phthiraptera, Philopteridae) infecting the domestic pigeon Columba livia from Saudi Arabia.

The taxonomy of the order Phthiraptera is unstable and still problematic to researchers. Most of the current taxon classifications are mainly based on morphological features. Campanulotes bidentatus belongs to the chewing lice of the Philopteridae family that mostly parasitic on birds. There is a lack of sequence data and phylogenetic analyses on the family Philopteridae. In the current study, C. bidentatus was collected from the domestic pigeon Columba livia and identified morphologically and molecularly based on the mitochondrial cytochrome c oxidase subunit 1 gene (COI). The infection rate of the Campanulotes genus was approximately 58.82% in this study. Phylogenetic analysis based on the mt COI gene was informative for members of Philopteridae and the group taxon genera formed distinct clades. Future studies were recommended using the 16s rRNA to enhance the tree topology and obtain clear differentiation between genera.

Sarcocystis camelicanis increases interleukin (IL)-6 expression in one-humped camels (Camelus dromedarius) from Riyadh and Al Qassim, Saudi Arabia.

Sarcocystis spp. are intracellular protozoan parasites with an intermediate-definitive host life cycle based on a prey-predator relationship. Sarcocystis infection is common among different vertebrates including humans. The pathogenicity of Sarcocystis spp. is of varied significance including a possible lethal effect for the host. The goal of the present study was to investigate the inflammatory activity of Sarcocystis spp. in different organs of naturally infected camels. The tongue, esophagus, heart, diaphragm, and skeletal muscles were collected from 50 camels, and the tissues assessed for the presence of Sarcocystis spp. by macroscopic examination, light microscopy, and transmission electron microscopy (TEM). Moreover, expression of the interleukin (IL)-6 was analyzed using reverse transcriptase quantitative polymerase chain reaction (qPCR). Microscopic Sarcocystis spp. cysts were found in camels. TEM identified the cysts as Sarcocystis camelicanis (S. camelicanis). Sarcocystis infection increased inflammation by stimulation of IL-6 expression in different organs of the camels, particularly in those from the Al-Qassim region.

A rapid and simple single-step method for the purification of Toxoplasma gondii tachyzoites and bradyzoites.

This study describes a simple method for the large-scale isolation of pure Toxoplasma gondii tachyzoites and bradyzoites. T. gondii tachyzoites were obtained from infected human foreskin fibroblasts (HFFs) and peritoneal exudates of mice, while tissue cysts containing bradyzoites were collected from chronically infected mice. Harvested cells and brain tissues were incubated in Hanks balanced salt solution (HBSS), containing 0.25% trypsin and 0.5% taurodeoxycholic acid (TDC) for 5 min. Subsequent washes in phosphate buffered saline (PBS) were conducted, and the cell viability of the preparations was good, as determined by flow cytometry and ability to reinfect HFF cells and propagate in mice. The purification procedure allowed for a rapid preparation of pure T. gondii tachyzoites and bradyzoites in sufficient quantity that can be used for downstream procedures. The advantage of the new method is that it is convenient and inexpensive.

Prevalence of eimeriosis in the one-humped camels (Camelus dromedarius) from Riyadh and Al-Qassim, Saudi Arabia.

BACKGROUND: The one-humped camels are economically important for several countries in Africa, Asia, and the Arabian Peninsula. Coccidiosis causes significant economic impact. Studies on coccidian parasite species causing such infections are limited. The present study aimed to carry out a survey of Eimeria spp. in camels from Riyadh and Al-Qassim, Saudi Arabia. METHODS: A total of 209 fecal samples from Camelus (C.) dromedarius slaughtered in West Abattoir in Riyadh and Onaizah Modern abattoir in Al-Qassim were collected. Samples were examined by flotation methods and oocyst sporulation. RESULTS: Of the 209 examined fecal samples, 75 were positive for Eimeria spp..The prevalence of oocysts in Riyadh and Al-Qassim were 33.89% (40/118) and 38.46% (35/92), respectively. The prevalence in young male camels was 41.02% (32/78) and 39.62% (21/53), respectively and in adult males was 19.35% (6/31) and 36% (9/25), respectively. Adult females displayed a prevalence of 22.22% (2/9) and 38.46% (5/13) in Riyadh and Al-Qassim, respectively. Three Eimeria spp. were identified; E. cameli, E. rajasthani, and E. pellerdyi. The presence of E. pellerdyi is considered the first record in Saudi Arabia.

Identification of Sarcocystis Spp. in One-humped Camels (Camelus dromedarius) from Riyadh and Dammam, Saudi Arabia, via Histological and Phylogenetic Approaches.

Sarcocystis (S.) spp. are intracellular protozoan parasites that infect birds and animals, resulting in substantial commercial losses. Sarcocystis spp. have an indirect life cycle; canines and felines are known to act as final hosts, and numerous domestic and wild animals act as intermediate hosts. The presence of sarcocysts in camel meat may diminish its commercial quality. There is limited knowledge regarding the taxonomy and diagnosis of Sarcocystis spp. that infect camels in Saudi Arabia. In this study, transmission electron microscopy (TEM) revealed S. cameli and S. camelicanis (camelicanis) in Camelus (C.) dromedarius. This is the first report of S. camelicanis in Saudi Arabia and is considered a significant finding. Based on cytochrome c oxidase subunit I gene (COX1) sequences, two samples of Sarcocystis spp. isolated from C. dromedarius in Riyadh and Dammam were grouped with S. levinei hosted by Bubalus bubalis in India, S. rangi hosted by Rangifer tarandus in Norway, S. miescheriana hosted by Sus scrofa in Italy and S. fayeri hosted by Equus caballus in Canada. The sequences obtained in this study have been deposited in GenBank.

The role of honeybee pollen as a natural source of antioxidants in the in vitro maturation medium of sheep oocytes and its effect on gene expression.

This study aimed to investigate the effect of honeybee pollen as an antioxidant source in a maturation medium of sheep oocytes on the in vitro maturation rate, glutathione concentration, and gene expression. To our knowledge, this study might be the first of its kind in this field. Sheep oocytes were cultured in vitro with honeybee pollen at four different concentrations (0.0, 1.0, 10.0, and 50.0 µg/ml). The results indicated that the ratio of oocytes that reached metaphase II stage was higher in the honeybee pollen-treated groups than in the control group (p ≤ 0.05). The reduced glutathione (GSH) mean content of matured oocytes was 9.85 nmol/25 oocytes, when honeybee pollen was added to the in vitro maturation (IVM) medium at a concentration of 1.0 µg/ml, compared with 5.84 and 4.44 nmol when using 10.0 and 50.0 µg/ml honeybee pollen, respectively. On the other hand, there was no significant difference in glutathione concentration between the control and 1.0 µg/ml honeybee pollen groups. Expression of candidate genes (GDF-9, BAX, Cyclin B, C-MOS, and IGF1) was upregulated in oocytes cultured with honeybee pollen when compared with oocytes cultured without honeybee pollen. In conclusion, the addition of honeybee pollen at a concentration of 1.0 µg/ml to IVM medium improved the in vitro maturation rate of sheep oocytes, increased the glutathione concentration, and improved gene expression.

Use of black seed (Nigella sativa) honey bee to improve sheep oocyte maturation medium.

Sheep are important livestock and a source of milk, meat, and wool globally. The increasing demand for animal protein requires increased productivity in sheep. In vitro fertilization and maturation can improve sheep productivity. The aims of this study were to evaluate the effects of honey bee addition as a supplementation medium on in vitro maturation improvement, gene expression of matured sheep oocytes, and determine the optimum concentration from honey bee for in vitro maturation of sheep oocytes. Cumulus oocyte complexes were obtained from the ovaries of slaughtered female sheep. Grade A and B oocytes were cultured for 24 h in medium without honey bee (control, G1) or medium supplemented with 5% (G2), 10% (G3), or 20% (G4) honey bee. Oocyte maturation rate, glutathione concentration, and the expression of candidate genes (GDF-9, BAX, Cyclin B, C-MOS, IGF1) were determined in the matured oocytes. The maturation rate of sheep oocyte was better in the presence of 5% and 10% honey bee; the mean number of oocytes in metaphase II stage was higher than that in G1 and G4 groups. Glutathione concentration was highest in G2 (10.93 ± 0.57). In general, gene expression levels were similar in G2 and G3, which were greater that in G1 and G4. In conclusion, the optimal concentration of black seeds honey bee that can be added to the maturation medium is 5% to obtain the highest mean MII and glutathione concentration values, and to improve gene expression in in vitro matured sheep oocytes.

Silver Nanoparticles Biosynthesized With Salvia officinalis Leaf Exert Protective Effect on Hepatic Tissue Injury Induced by Plasmodium chabaudi.

Malaria is an important health problem in subtropical and tropical areas around the world. Infection with protozoan parasites of the Plasmodium genus, which grow inside host erythrocytes, causes malaria and may lead to morbidity and mortality. Liver tissue plays an important role in the pathogenesis of malaria and is closely involved in parasitic pre-erythrocytic development. Numerous published studies have demonstrated that the liver is not only the source of Plasmodium parasites prior to erythrocytic growth but is also a primary immune effector toward the blood stage of the malaria life cycle. Despite efforts to improve antimalarial drugs and vaccines, Plasmodium species that cause severe malaria are being detected increasingly frequently in endemic regions. In this study, Salvia officinalis (S. officinalis) leaf extract was employed to synthesize silver nanoparticles (Ag-NPs). This method is eco-friendly and represents a single-step technique for the biosynthetic process; therefore, it has attracted considerable attention. Accordingly, we biosynthesized Ag-NPs with extract of the S. officinalis leaf and examined the antimalarial activity of these nanoparticles in a murine model of Plasmodium chabaudi malaria (P. chabaudi malaria). Forty mice were chosen and classified into four types: infected group, healthy control, pretreated mice infected after treatment with 50 mg/kg of S. officinalis leaf extract-biosynthesized Ag-NPs for two weeks, and post-treated mice infected before treatment with 50 mg/kg of S. officinalis leaf extract-biosynthesized Ag-NPs (administered daily for 7 d). In this study, both pre-treatment and post-treatment with Ag-NPs produced a substantial reduction in parasitemia relative to the infected group. We investigated the antiplasmodial and hepatoprotective effects of S. officinalis leaf extract-biosynthesized Ag-NPs on P. chabaudi-induced inflammation and hepatic oxidative stress markers.

Forensic insects attracted to human cadavers in a vehicular environment in Riyadh, Saudi Arabia.

Insect fauna attracted to cadavers at the crime scene can be identified and used to estimate the post-mortem interval (PMI). In the current study, insects associated with two human cadavers in a vehicular environment were collected and analysed. The first cadaver was found five days' post mortem in a garage. The second cadaver was found in a car ten days after his death. The weather conditions were obtained from the nearest weather station located to the scenes of the death. During the study, six adults, 32 larvae and egg batches were collected from case 1 and identified as Chrysomya albiceps Wiedemann (Diptera: Calliphoridae). From the second case, two larvae of Megaselia scalaris Loew (Diptera: Foridae) and seven larvae of Musca domestica L. (Diptera: Muscidae) were identified. mPMI ranged from five to twelve days. This information expands the knowledge on the insect fauna in the vehicular environment, which could be used to assist estimation of the PMI.

Anti-Toxoplasma activity of silver nanoparticles green synthesized with Phoenix dactylifera and Ziziphus spina-christi extracts which inhibits inflammation through liver regulation of cytokines in Balb/c mice.

Toxoplasmosis constitutes a global infection caused by oblige intracellular apicomplexan protozoan parasite Toxoplasma gondii Although often asymptomatic, infection can result in more severe, potentially life threatening symptoms particularly in immunocompromised individuals. The present study evaluated the anti-Toxoplasma effects in experimental animals of silver nanoparticles synthesized in combination with extracts of natural plants (Phoenix dactylifera and Ziziphus spina-christi) as an alternative method to standard sulfadiazine drug therapy. Liver functions estimated by and AST and ALT were significantly increased in T. gondii-infected mice compared with the control group as well as hepatic nitric oxide (NO), lipid peroxidation (LPO) levels and caused significant decrease in superoxide dismutase (SOD), catalase (CAT) and glutathione activities in the liver homogenates. Nanoparticles pretreatment prevented liver damage as determined by enzyme activity inhibition, in addition to significant inhibition of hepatic NO levels and significant elevation in liver SOD and CAT activities. Moreover, nanoparticle treatment significantly decreased hepatic LPO and NO concentrations and proinflammatory cytokines but significantly boosted the antioxidant enzyme activity of liver homogenate. In addition, histological examinations showed distinct alterations in the infected compared with untreated control groups. Conversely, nanoparticles pretreatment showed improvement in the histological features indicated by slight infiltration and fibrosis, minimal pleomorphism and less hepatocyte and degeneration. Furthermore, nanoparticles treatment induced a reduction in immunoreactivity to TGF-ß and NF-κB in hepatic tissues. Therefore, the present study provides new insights into various natural plants that are used traditionally for the treatment of toxoplasmosis and other parasitic infections, which may be useful as alternative treatment option for T. gondii infections.