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Plant growth-promoting bacteria (PGPB), possess multifarious beneficial traits and facilitate plant growth by both direct and indirect mechanisms under hostile conditions. The objective of the current study was to evaluate the potential application of 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) producing plant-growth promoting endophyte bacterium (PGPEB) Kocuria rhizophila strain 14ASP (National Center for Biotechnology Information; NCBI) Accession Number: LFIY00000000.1) heavy metals and salt resistance tolerance under hostile environment. The in vitro study was conducted to evaluate plant growth promotion characteristics, heavy metal and salt resistance of the K. rhizophila under normal and stressful environments. The strain 14ASP revealed the plant-growth promoting and of AAC deaminase activities under abiotic stress environment. K. rhizophila showed significant tolerance against different heavy metals on LB (Luria Ber-tani) agar plates and broth medium supplemented with cadmium (Cd), (50 mg L−1), cooper (Cu), (50 mg L−1), nickel (Ni), (50 mg L−1), lead (Pb), (200 mg L−1) and chromium (Cr) (500 mg L−1), respectively. It also exhibited population density (OD = 0.31–0.45) at different concentrations of sodium chloride (NaCl) (0% 5%, 10%, and 15% w/v) in LB medium. The draft genome of strain 14ASP is 2.6 Mb and was assembled into 183 contigs with 2689,1 bp and a G + C content of 70% and 1,882,646 bp, encoded 2409 protein, 8 rRNAs and 46 tRNAs. Genome analysis identified genes, involved in hydrocarbon metabolism, heavy metal tolerance, biofilm formation, Indole Acetic Acid (IAA) and siderophore biosynthesis. All these properties were confirmed by in silico and draft genome analysis.

Flonicamid is a chordotonal modulator and novel systemic insecticide that has been used frequently for controlling a broad range of insect pests. The risk of flonicamid resistance was assessed through laboratory selection and determining inheritance pattern and cross-resistance potential to five insecticides in house fly, Musca domestica L. Very low to high flonicamid resistance in M. domestica populations was found compared with the susceptible strain (SS). A flonicamid-selected (Flonica-RS) M. domestica strain developed 57.73-fold resistance to flonicamid screened for 20 generations compared with the SS. Overlapping 95% fiducial limits of LC50 of the F1 and F1ǂ, and dominance values (0.87 for F1 and 0.92 for F1ǂ) revealed an autosomal and incomplete dominant flonicamid resistance. The monogenic model of resistance inheritance suggested a polygenic flonicamid resistance. The Flonica-RS strain displayed negative cross-resistance between flonicamid and sulfoxaflor (0.10-fold) or clothianidin (0.50-fold), and very low cross-resistance between flonicamid and flubendiamide (4.71-fold), spinetoram (4.68-fold), or thiamethoxam (2.02-fold) in comparison with the field population. The estimated realized heritability (h2) value of flonicamid resistance was 0.02. With selection mortality 40–90%, the generations required for a 10-fold increase in LC50 of flonicamid were 94–258 at h2 (0.02) and slope (3.29). Flonicamid resistance was inherited as autosomal, incomplete dominant, and polygenic in the Flonica-RS. Negative or very low cross-resistance between flonicamid and sulfoxaflor, clothianidin, flubendiamide, spinetoram, and thiamethoxam means that these insecticides can be used as alternatives for controlling M. domestica. These data can be useful in devising the management for M. domestica.

Coronaviruses (CoVs) infect a wide range of domestic and wild mammals. These viruses have a potential and tendency to cross-species barriers and infect humans. Novel human coronavirus 2019-nCoV (hCoV-19) emerged from Wuhan, China, and has caused a global pandemic. Genomic features of SARS-CoV-2 may attribute inter-species transmission and adaptation to a novel host, and therefore is imperative to explicate the evolutionary dynamics of the viral genome and its propensity for differential host selection. We conducted an in silico analysis of all the coding gene sequences of SARS-CoV-2 strains (n = 39) originating from a range of non-human mammalian species, including pangolin, bat, dog, cat, tiger, mink, mouse, and the environmental samples such as wastewater, air and surface samples from the door handle and seafood market. Compared to the reference SARS-CoV-2 strain (MN908947; Wuhan-Hu-1), phylogenetic and comparative residue analysis revealed the circulation of three variants, including hCoV-19 virus from humans and two hCoV-19-related precursors from bats and pangolins. A lack of obvious differences as well as a maximum genetic homology among dog-, cat-, tiger-, mink-, mouse-, bat- and pangolin-derived SARS-CoV-2 sequences suggested a likely evolution of these strains from a common ancestor. Several residue substitutions were observed in the receptor-binding domain (RBD) of the spike protein, concluding a promiscuous nature of the virus for host species where genomic alternations may be required for the adaptation to novel host/s. However, such speculation needs in vitro investigations to unleash the influence of substitutions towards species-jump and disease pathogenesis. Electronic supplementary material The online version of this article (10.1007/s11033-020-05879-5) contains supplementary material, which is available to authorized users.

We investigated the extent and pattern of damage in wheat caused by Tribolium castaneum Herbst (Coleoptera: Tenebrionidae), Trogoderma granarium Everts (Coleoptera: Dermestidae) and Rhyzopertha dominica F. (Coleoptera: Bostrichidae) over a period of 75 d. Damage parameters included numbers of damaged grains, their weight; numbers of undamaged grains, their weight; frass weight and percent weight loss. Results showed after 75 d, maximum damaged grains were 125.17 ± 22.88 by R. dominica followed 80.50 ± 6.91 by T. granarium and least were 20.00 ± 2.61 after 50 d by T. castaneum. For R. dominica and T. granarium damage to grains increased with time (P 0.05). Maximum percent weight loss was 7.02 ± 1.87 g by R. dominica followed by 2.89 ± 0.33 g by T. granarium and least 0.65 ± 0.08 g by T. castaneum. Maximum dead insects percentage was 82% after 25 d for R. dominica, 59.67 ± 3.44 for T. granarium after 50 d and 100.00 ± 0.00 for T. castenum after 50 or 75 d compared to other time periods (P

A pot trial was established to evaluate the response of different potato genotypes to drought stress. Two potato genotypes (Demon and Hopehely) were exposed to two water levels (80% and 50% water holding capacity). The trial was replicated 16 times and 4 replications were harvested at 36 DAS, 54 DAS, 72 DAS, and 90 DAS. The results revealed that drought significantly reduced the growth and plant development of Demon (G1). Hopehely (G2) produced a higher yield in control as well as drought condition. Tuber yield positively correlated with tubers number, and root weight that was significantly higher in Hopehely under both experimental conditions. Hopehely produced a higher number of tubers (18) than Demon whereas Demon produced 11 tubers per plant. Moreover, under drought conditions, the relative water content of leaves and nitrogen content in foliage increased in Hopehely while decreased in Demon. Drought stress caused a 40% reduction in plant height and a 24.3% reduction in the number of leaves in Demon that was significantly higher than the 11% plant height reduction and 9.1% leaf count reduction in Hopehely. It was observed that the morphology of Hopehely (producing dwarf plants, fewer leaves, maintaining water content of leaves, producing more tubers) helped it to be a better drought-tolerant genotype compared to Demon.

Helicobacter pylori and HCV are the most prevalent pathogens worldwide associated with high rates of morbidity and mortality. This study aimed to determine the prevalence of H. pylori in HCV infected patients and compares changes in liver function tests (LFTs) of both HCV mono-infected and HCV/H. pylori co-infected patients. Eighty-seven participants participated in this study who were first screened for HCV using GeneXpert technology followed by H. pylori antigen stool test. A complete blood count (CBC) analysis and LFTs were performed to determine the impact of HCV and H. pylori on liver enzymes. Our findings suggested that 45% of patients had HCV mono-infection, whereas 55% were co-infected with both HCV and H. pylori. Furthermore, we also observed that ALT, AST, and ALP levels were significantly elevated in HCV/H. pylori co-infected patients as compared to HCV mono-infected patients. To the best of our knowledge, this is the first study to report the prevalence of H. pylori in HCV infected subjects of Lahori population

Introduction Helicobacter pylori (H. pylori) infection is prevalent worldwide. H. pylori therapies' adverse effects can contribute to noncompliance among patients. This study aimed to assess the association between compliance to H. pylori eradication therapy and adverse effects using various drug regimens. Method We conducted an observational study from September 2017 to February 2020 in two tertiary care hospitals in patients with dyspeptic symptoms. H. Pylori detection was done by histopathological examination of gastric mucosa during upper gastrointestinal endoscopy or stool for H. pylori antigen. Patients with positive results were randomly assigned one of the nine different regimens consisting of a combination of proton pump inhibitors along with at least two antibiotics. The antibiotics used in different combinations were amoxicillin, clarithromycin, metronidazole, doxycycline, levofloxacin, and bismuth sulfate. The treatment groups received standard triple therapy with and without probiotics, sequential, concomitant, levofloxacin-based triple therapy, or sequential and bismuth-based quadruple treatments. All treatments were given for two weeks. At the end of the treatment period, patients were interviewed about completing treatment and any adverse effects they may have experienced during therapy. Data were analyzed using IBM Statistical Package for the Social Sciences (SPSS) Statistics for Windows, Version 22.0 (Armonk, NY: IBM Corp.). Results A total of 250 patients were included in the study (62% males, 38% females) with a mean age of 37 years ± 13 years (range 12-84 years). Most patients completed the treatment regimen (80.4%), and 19.6% did not complete treatment because of adverse effects (p

Banana (Musa spp.) is one of the most widely grown and consumed fruits in Pakistan and all around the world due to their distinct aroma and taste. In 2018, anthracnose symptoms were observed on banana fruit harvested from different plantations of Sindh- a major banana producing Province of Pakistan. Approximately, 25% of banana fruit collected from different plantations were infected. The symptoms consisted of small brown to reddish-brown spots on the fruit surface and then became sunken lesions as the disease progressed. To identify the pathogen, infected tissues (5 mm in diameter) from the margin of the lesions were surface sterilized by dipping in 1% sodium hypochlorite (NaOCl) for 2 min, 70% ethanol for 30 s, and then rinsed twice with sterile distilled water, plated onto potato dextrose agar (PDA), and incubated at 27°C for 5 days with 12 h light and darkness cycle. Colonies with a similar pattern were consistently isolated and all colonies were sub-cultured using the single-spore method. Colonies first appeared with white colored mycelium and later turned to dark gray. Conidia produced in acervuli were cylindric, hyaline, straight, and aseptate, with both ends rounded. Conidia measured 14.0 ± 0.5 × 3.4 ± 0.6 μm. Conidiomata were dark brown and spherical. On the basis of morphological characterization, the pathogen was identified as Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. (Weir et al. 2012). Two independent isolates (PDL2031 and PDL2032) were used for further genetic analysis. The internal transcribed spacer (ITS) region and chitin synthase 1 (CHS-1) gene were amplified from genomic DNA using primer pairs of ITS1/ITS4 and CHS-79F/CHS-345R, respectively (White et al. 1990; Damm et al. 2012). The GenBank accession numbers (MW493198, MW504711 for ITS and MW530421, MW530422 for CHS-1) of the sequences exhibited 99% to 100% identity to multiple sequences of C. gloeosporioides. To conduct a pathogenicity test, 10 healthy fruits were selected and surface sterilized with 70% ethanol followed by a wash of sterilized water. The fruits were stabbed with a sterile needle and a drop of 20 µl of spore suspension (106 spores/ml) was placed on each wound independently. Meanwhile 10 fruits inoculated with sterile water were treated as controls. The fruits were incubated at 27°C with 90% relative humidity for 10 days. Inoculated fruits exhibited symptoms similar to the original infection. No visible lesions appeared on control fruit. C. gloeosporioides was successfully reisolated from the inoculated fruit, confirming Koch's postulates. Anthracnose of banana is known to be caused by C. musae, C. gloeosporioides, C. siamense, C. tropicale, C. chrysophilum, C. theobromicola, and C. scovillei (Kumar et al. 2017; Peres et al. 2001; Vieira et al. 2017; Zakaria et al. 2009; Zhou et al. 2017). To our knowledge, this is first report of anthracnose of banana caused by C. gloeosporioides in Pakistan. The new disease primarily reduces the quality and yield of Banana. Effective measures should be taken to manage this disease.

The clinicopathological effects of E. coli infected Psittaculakrameri (ring necked parrots) supplemented with N. sativa seeds in feed were evaluated. Faecal samples from “psittacine” species parrots were collected and E. coli was isolated for cultural, morphological, biochemical and in-vitro studies. A total of 27 parrots were selected and divided into 03 groups (A= control, B= no treatment and C= treatment+ infection) having 09 parrots in each and were fed with N. Sativa seeds @100mg/kg from 3rd day. The parrots of group B and C were challenged with 0.1ml pathogenic E. coli @106CFU/ml/bird subcutaneously on day 9th. Clinical signs, mortality and morbidity rates were observed during the trial. For haematology, histopathology and CRP in sera, three birds were slaughtered weekly from each group. Significant decreased in feed consumption, haematological values like red blood cells, haemoglobin and pack cell volume while significant increase in clinical signs, morbidity, mortality, liver, heart, kidneys weights and white blood cell were recorded in group B. C-Reactive protein values was significantly decreased in group C as compared to group B while degenerative changes were seen in liver, heart and kidneys of group B.

Mungbean yellow mosaic virus (MYMV) is an important constraint in successful production of mungbean (Vigna radiata L.) in many countries, including Pakistan. The MYMV spreads by insect vector whitefly (Bemisia tabaci Gennadius). The use of resistant cultivars is the most effective management tactics for MYMV. Twenty mungbean varieties/lines were screened against insect vector of MYMV under field condition in the current study. Resistance levels for varieties/lines were assessed through visual scoring of typical disease symptoms. Furthermore, the impacts of two insecticides ‘Imidacloprid’ and ‘Thiamethoxam’ and two plant extracts, i.e., neem (Azadirachta indica), and Eucalyptus (Eucalyptus camaldulensis) were tested on the suppression of whitefly. Field screening indicated that none of the tested varieties/lines proved immune/highly resistant, while significant variations were recorded among varieties/lines for resistance level. All varieties/lines were systemically infected with MYMV. The varieties ‘AARI-2006’ and ‘Mung-14043’ were considered as resistant to MYMV based on visual symptoms and the lowest vector population. These varieties were followed by ‘NM-2006’ and ‘NL-31’, which proved as moderately resistant to MYMV. All remaining varieties/lines were grouped as moderately to highly susceptible to MYMV based on visual symptoms’ scoring. These results revealed that existing mungbean germplasm do not possess high resistance level MYMV. However, the lines showing higher resistance in the current study must be exploited in breeding programs for the development of resistant mungbean varieties/lines against MYMV. Imidacloprid proved as the most effective insecticide at all concentrations to manage whitefly population. Therefore, use of the varieties with higher resistance level and spraying Imidacloprid could lower the incidence of MYMV.

Novel mutant camelina has become a crop of interest inspired by its short growing season, low harvesting costs and high oil composition. Despite those advantages, limited research has been done on novel mutant lines to determine applicability for biodiesel production. Jatropha is an extremely hardy, frugal and high oil yielding plant species. The major aim of the present study was not only to compare biodiesel production from jatropha and camelina but was also to test the efficacy of camelina mutant lines (M6 progenies) as superior feedstock. The biodiesel yield from camelina oil and jatropha oil was 96% and 92%, respectively. The gas chromatographic analysis using flame ionization detector (GC-FID) showed that mutant camelina oil biodiesel sample contain major amount of oleic acid (46.54 wt%) followed by linolenic acid (20.41 wt%) and linoleic acid (16.55 wt%). Jatropha biodiesel found to contain major amount of oleic acid (45.03 wt%) followed by linoleic acid (25.07 wt%) and palmitic acid (19.31 wt%). The fuel properties of produced biodiesel were found in good agreement with EN14214 and ASTM D6751 standards. The mutant camelina lines biodiesel have shown comparatively better fuel properties than jatropha. It has shown low saponification value (120.87–149.35), high iodine value (130.2–157.9) and better cetane number (48.53–59.35) compared to jatropha biodiesel which have high saponification value (177.39–198.9), low iodine value (109.7–123.1) and lesser cetane number (47.76–51.26). The results of the present student of utilizing novel mutant camelina lines for biodiesel production are quite promising and are helpful in turning out the outcomes of the previous studies suggesting that C. sativa biodiesel presents serious drawbacks for biodiesel applications.

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a frustrating and often debilitating disease. Current studies have shown that Traditional Chinese Medicine (TCM) can improve patients' quality of life and alleviate CP/CPPS symptoms. In this study, the efficacy of Hedyotis diffusa Willd aqueous extraction in experimental autoimmune prostatitis (EAP) mice models was revealed. The C57BL/6 mice were randomly assigned to three groups. Except for the control group, all other groups were subcutaneously injected with 0.2 ml emulsion of T2 peptide, on day 0 and day 14, for inducing EAP models. After the EAP modelling, oral saline was given to the model group, while the H. diffusa group was treated with aqueous extract of H. diffusa Willd. Micturition habits and withdrawal response frequencies were measured. Haematoxylin and eosin staining and immunohistochemistry were used to investigate inflammatory cell infiltration and TNF-α in the prostate tissue respectively. TNF-α levels in the serum were evaluated by ELISA. The H. diffusa Willd aqueous extraction considerably reduced the urine spots number and increased the pain threshold in H. diffusa group. H. diffusa group showed significantly reduced inflammatory lesion and inflammatory cell infiltration than the model group. The levels of TNF-α in H. diffusa group were considerably reduced.

Renessans is an iodine complex which has proven in vitro antiviral activity including Anti-SARS-CoV-2 activity. The present study was designed to determine its efficacy against SARS-CoV-2 in monkeys (Rhesus macaque). A total of 14 monkeys were divided into four groups: A) Prophylactic group (n=03), (B) Treatment group (n=03), (C) infection control group (n=04) and (D) negative control group (n=04) and were housed in BSL-3 Animal facility while group D was housed at another animal house. Group A was administered with Renessans @ 2.85 mg/7 kg from 5 days prior to the infection to 08 days post infections (DPI). Group B was administered with Renessans from 03-08 DPI @ 2.85 mg/7 kg. Group C was administered with WIF only. The infection @ 2 × 106 TCID of SARS-CoV-2 was given to all group monkeys through intranasal and oral route under anesthesia. Nasal swab samples (at different times) and fecal matter on daily basis were collected for the detection of SARS-CoV-2 through real-time quantitative PCR. Three monkeys (one from each of group A, B and C) were euthanized at 07 DPI to determine the gross pathological lesions and SARS-CoV-2 detection from internal tissues. Nasal swabs from all the monkeys from group A, B and C were positive for SARS-CoV-2 at 02 and 07 DPI (Day 05 of treatment). At 14 DPI, all (100%) nasal swabs from group A were negative for SARS-CoV-2 while 50% and 100% were positive from group B and C, respectively. At 21 DPI, monkeys from group B were negative and all in group C were still positive for SARS-CoV-2. Similarly, fecal matter of monkeys in group A and B was returned negative in significantly lesser time as compared to monkeys from infection control group. Based on these research findings it is concluded that the Renessans has in-vivo SARS-CoV-2 activity and may result in early clearance of SARS-CoV-2. Therefore, a clinical trial of the drug in COVID-19 patients may reveal its anti-COVID-19 potential.

Introduction: Cystic echinococcosis, caused by Echinococcus granulosus, is a neglected zoonosis that affects humans and livestock. This sero-survey was designed for the first time in Pakistan to assess the exposure of butchers to E. granulosus as there was no previous report in the country for this occupational group. Methodology: Blood samples were collected from registered butchers (n = 364) in five different slaughterhouses in Faisalabad and Bahawalnagar Districts. Sera were tested for anti-Echinococcus granulosus IgG with a commercially available ELISA kit (specificity, 100%; sensitivity, 97%). Results: Overall, seroprevalence was 9.61% (35/364). Butchers >30 years of age (10.34%), those involved in small ruminants butchery (11.70%), >10 years’ experience (10.04%), formal education level up to middle standard (10.28%), contact with dogs (12.71%), improper/unhygienic disposal of dog feces (11.87%), and those unaware of the consequences of eating with unwashed hands (13.80%) were more seropositive with significant statistical differences (p < 0.05). Variables like previous cyst encounter, no knowledge of zoonoses and/or cystic echinococcosis, living in rural areas and the presence of stray/feral dogs in surroundings did not show any significant association (p > 0.05) with seroprevalence in butchers. The binary logistic regression model also showed a statistically significant relationship (p < 0.05) for all risk factors found statistically significant (p < 0.05) in the univariate analysis. Conclusions: This study shows high prevalence of cystic echinococcosis among butchers in Pakistan and underscores the need for educating native slaughterhouse personnel on cystic echinococcosis. It also serves as a global warning, especially in developing countries.

Artificially boosting body's immune response is one of the most exciting, effective and promising advancements in the treatment of cancers. Cancer immunotherapeutics consist of variety of treatment approaches such as cytokine therapy, adoptive T-cell transfer therapy, and antibodies that stimulate innate and adoptive immune responses. In addition to this, development of HPV vaccine has paved way toward the development of other cancer vaccines. Checkpoint blockade inhibitors, for example, anti-programmed cell death protein 1 and anti-cytotoxic T-lymphocyte-associated antigen-4, chimeric antigen receptor T-cell therapy and monoclonal antibodies are emerging as other major breakthroughs that are highly effective against cancer. This review addresses the current status of immunotherapeutic strategies against cancer and provides baseline data for future research.

Sesame (Sesamum indicum L.) commonly known as 'til' is the most ancient and widely grown oilseed crop of Pakistan. During 2020, field survey conducted in various research fields of Faisalabad (31°26'00.2″N, 73°04'25.4″E) revealed the occurrence of characteristic leaf blight disease with an incidence of 10 to 13%. The symptoms were characterized by yellow-brown and irregular lesions. At later stages, the lesions expanded and the affected leaves turned grayish to dark-brown and finally became wilted. Symptomatic leaves (both the diseased and healthy tissue) were cut into approximately 2 × 2 mm pieces, surface sterilized with 1% sodium hypochlorite for 30 s, 70% ethanol for 30 s, and finally, three times in sterile distilled water prior to culturing on Potato Dextrose Agar (PDA) and incubated at 25 °C under a 12-h photoperiod for 7 days. To obtain pure cultures, hyphal tips of growing mycelia from leaf tissues were carefully isolated and transferred onto fresh PDA plates. Fungal colonies of 11 isolates were initially white, becoming light to dark-gray. The conidia were black, spherical to subspherical, and single-celled (12 to 14 × 18 to 20 μm) in diameter, which were borne on a hyaline vesicle at the tip of the conidiophore. Further, to identify the pathogen to the species level, genomic DNA was extracted using a modified CTAB protocol described by (Guo et al. 2000). The internal transcribed spacer (ITS) region of the ribosomal DNA and translation elongation factor 1-alpha (TEF1-α) gene were amplified using ITS1/ITS4 (White et al. 1990) and EF1-728F/EF1-986R primer sets (Carbone and Kohn 1999), respectively. The sequences were submitted to GenBank (accession no. MW287214 for ITS and MW325222 for TEF1. The sequences comparison revealed 99% and 100% similarity to multiple sequences of N. sphaerica (GenBank accessions KX834822 and MN995332). On the basis of cultural features, conidial morphology and molecular data, the fungus was identified as Nigrospora sphaerica (Sacc.) Mason (Wang et al. 2017; Chen et al. 2018). To test the pathogenicity, fresh and healthy leaves of ten 6-week-old growth stage sesame plants were spray inoculated with a conidial suspension (105 conidia/ml), collected from a 7-day-old culture on PDA. In addition, 10 plants sprayed with sterile distilled water served as controls. Inoculated plants were covered with polyethylene bags to maintain high humidity and kept at 28°C, and observations were made at regular intervals. After 8-10 days of inoculation, leaves developed blight symptoms similar to those observed on naturally infected leaves, whereas control leaves remained asymptomatic. The pathogen was re-isolated from the inoculated leaves, and its identity was confirmed by morphological and molecular (ITS and TEF1) means, thus fulfilling Koch's postulates. N. sphaerica is distributed on a wide range of hosts and has been reported from different host genera including monocotyledonous and dicotyledonous (Wang et al. 2017). Previously, N. sphaerica has been reported to cause leaf blight of Cunninghamia lanceolata in China (Xu and Liu, 2016). To the best of our knowledge, this is the first report of Nigrospora sphaerica as the causal agent of leaf blight of sesame in Pakistan. Because sesame is an important oilseed crop of Pakistan, appropriate disease management practices should be developed and implemented.

Micronutrients play a vital role in the growth and productivity of cotton crop. A study was carried out to access the exogenous application of micronutrients on growth and yield of cotton crop. The experiment was comprised of nine treatments as T0 (control), T1 (Fe chelated), T2 (B), T3 (Mo), T4 (CuSo4 + ZnSo4 + MnSo4), T5 (CuSo4 + ZnSo4 + MnSo4 + Fe chelated), T6 (CuSo4 + ZnSo4 + MnSo4 + B), T7 (CuSo4 + ZnSo4 + MnSo4 + Mo) and T8 (CuSo4 + ZnSo4 + MnSo4 + Fe chelated + B). Data on different growth attributes showed that there was significant positive increase with the application of micronutrients. Leaf area was increased after applying micronutrients at 99 days after sowing (DAS) and then a decreasing trend was observed. Chlorophyll contents were increased at 81 DAS and then decreased towards the final harvest. Similarly, different yield components showed that seed cotton yield were significantly increased with the application of Fe, B, Mo, Zn, Cu and Mn compared to control treatment. Earliness index, mean maturity date and production rate index were increased significantly after combined use of foliar spray of Zn, Cu, Mn and Mo. Os micronutrientes desempenham um papel vital no crescimento e produtividade da cultura do algodão. Um estudo foi realizado para acessar a aplicação exógena de micronutrientes no crescimento e produção decultura de algodão. O experimento foi composto de nove tratamentos como T0 (controle), T1 (Fe quelatado), T2 (B), T3 (Mo), T4 (CuSo4 + ZnSo4 + MnSo4), T5 (CuSo4 + ZnSo4 + MnSo4 + Fe quelatado) T6 (CuSo4 + ZnSo4 + MnSo4 + B), T7 (CuSo4 + ZnSo4 + MnSo4 + Mo) e T8 (CuSo4 + ZnSo4 + MnSo4 + Fe quelados + B). Dados sobre diferentes atributos de crescimento mostraram aumento significativo positivo com a aplicação de micronutrientes. A área foliar foi aumentada após aplicação de micronutrientes aos 99 dias após a semeadura (DAS), observando - se, então, uma tendência decrescente. Os teores de clorofila foram aumentados em 81 DAS e depois diminuíram para a colheita final. De forma semelhante, diferentes componentes de rendimento mostraram que o rendimento de algodão de sementes aumentou significativamente com a aplicação de Fe, B, Mo, Zn, Cu e Mn em comparação com o tratamento de controlo. O índice de precocidade, a data média de maturidade eo índice de taxa de produção aumentaram significativamente após o uso combinado de pulverização foliar de Zn, Cu, Mn e Mo.

Food production and waste recycling are the two major issues faced globally with rapidly increasing population. Recycling organic wastes to crop amendments could be a possible solution to these issues. Earthworms transfer organic waste to compost, which is used to grow crops and increase crop productivity. This study assessed the impact of vermicompost produced from the residues of six desert plant species, i.e., (Ziziphus mauritiana, Aerva javanica, Calligonum comosum, Sacchrum benghalens, Calligonum polygonoides and Prosopis cineraria) combined with farmyard manure (5 t ha-1) on growth, yield and photosynthetic activity of maize crop. Earthworm species Eisenia fetida (Savigny, 1826) was used to prepare vermicomposting of all tested plant species. The desert species were collected from natural habitats, chopped, dried, mixed with FYM and then earthworms were released to prepare the vermicompost. The earthworms were excluded twenty days after release and resultant was considered as compost and used in the experiment. Results revealed that application of P. cineraria vermicompost resulted in the highest plant height (75.33 cm), stem diameter (22.66 mm), cob length (17.66 cm), number of grains/cob (374.67), 1000-grain weight (260.41 g) and grains yield (3.20 t/ha). Application of P. cineraria vermicompost resulted in the highest uptake of macronutrients, i.e., N (91.01%), P (22.07%), K (80.41%), micronutrients, i.e., Fe (19.07 ppm), Zn (40.05 ppm), and phenolic contents (150). Application of P. cineraria vermicompost also resulted in the highest quantum photosynthetic yield (0.42 mole C/mole of photon), chlorophyll florescence (355.18 moles of photon m-2s-1) and electron transport rate (310.18 micro mole m-2s-1). It is concluded that vermicomposting has the potential to improve growth and yield of maize crop. Particularly, application of vermicompost obtained from P. cineraria can be used to improve the growth and yield of maize crop. Nonetheless, field trials are necessary for a wide scale recommendation.

Pomegranate (Punica granatum L.) is a non-climacteric and a favorite fruit of tropical, sub-tropical and arid regions of the world. During a survey in autumn 2019, leaf lesions were observed on plants (cv. Kandhari) in different orchards of Muzaffargarh (30°4'27.7572″ N, 71°11'4.7544″ E), a major pomegranate-producing region in Punjab Province. Disease incidence ranged from 17 to 20%. Leaf lesions were initially small (1 to 3 mm in diameter), round, purple or reddish-brown, scattered spots. At later stages, spots increased in size and the centers of mature lesions became dark red or black with fungal sporulation. To isolate the pathogen, samples of leaf (5 × 5 mm) were cut from the junction of diseased and healthy tissue, surface disinfected in 75% alcohol for 30 s, sterilized with 6% sodium hypochlorite for 3 min, washed with sterile distilled water three times, air dried in laminar flow hood, and cultured on potato dextrose agar (PDA). After one week of incubation at 25 ± 2°C with a 12-h photoperiod, fungal colonies developed, which were initially white and became pale yellow with olivaceous green mycelium after 20 days. On PDA, ascomata were olivaceous green, with a papillate ostiole, globose or ovoidal to obovoidal (155 to 220 × 120 to 240 µm, n=50). Terminal and lateral setae were abundant, brown, and tapering toward the tips (4 to 6 µm, n=50). Asci were greenish and lemon-shaped (6 to 8 × 9 to 13.5 µm, n=50). Ascospores were limoniform and olivaceous gray-brown (10 to 11.5 × 7 to 9 µm, n=50). These morphological characteristics were consistent with the morphology of Chaetomium globosum (Lan et al. 2011; Wang et al. 2016). Genomic DNA was extracted from two isolates and identification of the pathogen was confirmed by amplification and sequencing of the internal transcribed spacer region (ITS) and the partial translation elongation factor 1-α (TEF1) gene using ITS1/ITS4 (White et al. 1999) and EF1-983F/EF1-2218R primers (Wang et al. 2016), respectively. The sequences of the PCR products were deposited in GenBank with accession numbers MW522514, MW522352 (ITS), and MW530423, MW530424 (TEF1). BLAST results of the obtained sequences of the ITS and TEF1 genes revealed 100% (513/513 bp) and 99.78% (927/929 bp) similarity with those of C. globosum in GenBank (ITS: KX834823 and KT898637, and TEF1: MG812564 and KC485028). To confirm pathogenicity, inoculum was prepared by harvesting conidia from 10-day-old culture grown in PDA. The surface-disinfected (70% ethyl alcohol, 30 s) leaves of ten 1-year-old seedlings (cv. Kandhari) were sprayed with a spore suspension (1×106 conidia/ml). Leaves of ten seedlings sprayed with sterile distilled water served as controls. All seedlings were covered with plastic bags and placed in a greenhouse at 26°C with 12 h photoperiod. After eight days, symptoms on inoculated leaves were similar to those observed in the orchards; no symptoms were observed on controls. The fungus was reisolated from all symptomatic tissues. C. globosum has been reported on Punica granatum (Guo et al. 2015), Cannabis sativa (Chaffin et al. 2020) and Brassica oleracea (Zhu et al. 2020). This is the first report of C. globosum causing leaf spot on pomegranate in Pakistan. This finding suggests a potential threat to pomegranate production in Pakistan and further studies should focus on effective prevention and control practices of this disease.

The research was conducted to evaluate in-vitro efficacy of numerous fungicides against Fusarium oxysporum f. sp. capsici. In present research, six treatments (T) viz. Carbendazim, Benomyl, Topsin-M, Difenoconazole, Nativo, and Alliete along with control, various concentrations (C), days (D), and their interactions, i.e., (T × C), (T × D), (C × D), and (T × C × D) were exploited in a laboratory through food poison technique. Alliete expressed maximum colony growth (1.93 cm) as compared to all other fungicides with respect to control. Interaction between treatments and concentration (T × C) exhibited maximum colony growth of all treatments (Carbendazim, Benomyl, Topsin-M, Difenoconazole, Nativo, and Alliete), i.e., 0.87, 1.23, 1.73, 2.20, 2.53, and 2.93 cm at 300 ppm as compared to 500 and 700 ppm concentrations, respectively. Similar trend was also observed concerning interaction between (fungicides × days) and (tested concentrations × days). Results of the present study revealed that among tested fungicides, Carbendazim at 700 ppm expressed significant reduction in fungal growth.

In this study, ALOS-2 PALSAR L-band dual-pol (HH and HV) synthetic aperture radar (SAR) images were used for above ground biomass (AGB) estimation of Shisham (Dalbergia sissoo) tree species in the ...

Background: The immunological response in COVID-19 individuals with positive SARS-CoV-2 PCR and its association with various demographic characteristics are inadequately characterized. Method: In this study we examined the presence of anti- severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antibodies among SARS-CoV-2 PCR positive patients and associated it further with demographic characteristics, ethnicity, symptoms, and the suspicion of virus exposure. Findings: Of the total 714 COVID-19 suspected individuals, 451 (63.16%) generated positive PCR results. The mean age of the enrolled patients was 38.94 (S.D, 16.86) years (range, 3-88 years). Total 337 (74.72%) were males, 267 (59.20%) were aged 3 to 40 years old, 323 (71.61%) were with Punjabi ethnicity and 347 (76.94%) had one or more COVID-19 specific symptoms. Of the 451 PCR positive cases, 349 (77.38%) tested positive for anti-SARS-CoV-2 antibodies and the observed seroconversion time was 14-28 days for total antibodies except one lady doctor whose antibodies reached to the detectable level at 8th month (56th day) of PCR positivity. The presence of total antibodies was detected in 25.1%, 49.8%, 74.5% and 74.7% of COVID-19 patients on day-14, 21, 28 and 56 respectively. No antibody was seen in 22.62% enrolled subjects till week-8th post PCR positivity. This seroconversion varied by age, ethnicity, and presence of COVID-19 specific symptoms. Antibody production was found significantly high in ages 40 years or below (OR 1.603; 95% CI 1.006-2.554), Punjabi ethnicity (OR 8.222; 95% CI 3.499-19.318), and presence of symptoms (OR 5.213; 95% CI 3.195-8.507). No significant difference was seen in seroconversion in male and female COVID-19 patients. Interpretation: Several important researches emerged from the current study: 1) As anti-SARS CoV-2 antibodies were detected in only 77.38% PCR positive cases and a significant number of COVID-22.62% did not develop anti-SARS-CoV-2 antibodies therefore, the serological tests may not be very useful for seroprevalence studies, if used the prevalence rate may be much higher.2) The time of antibody response is between 2-4 days in majority of the ARS-CoV-2 infected patients after positive PCR results however this time of seroconversion may take 8 weeks even in the absence of active SARS-CoV-2. 3) Public immunity to COVID-19 might be higher significantly than serologic tests indicate and may be a good news from a public health perspective. 4) The most common symptom in this region of the world is loss of smell and/or taste (52%) followed my myalgia (45%). Funding Statement: No funding received from any source. Declaration of Interests: None of the authors of this manuscript have any competing interest. Ethics Approval Statement: This research study was approved by GCMD Institutional Review Board (IRB).

Autism spectrum disorder (ASD) is a genetic heterogeneous neurodevelopmental disorder that is characterized by impairments in social interaction and speech development and is accompanied by stereotypical behaviors such as body rocking, hand flapping, spinning objects, sniffing and restricted behaviors. The considerable significance of the genetics associated with autism has led to the identification of many risk genes for ASD used for the probing of ASD specificity and shared cognitive features over the past few decades. Identification of ASD risk genes helps to unravel various genetic variants and signaling pathways which are involved in ASD. This review highlights the role of ASD risk genes in gene transcription and translation regulation processes, as well as neuronal activity modulation, synaptic plasticity, disrupted key biological signaling pathways, and the novel candidate genes that play a significant role in the pathophysiology of ASD. The current emphasis on autism spectrum disorders has generated new opportunities in the field of neuroscience, and further advancements in the identification of different biomarkers, risk genes, and genetic pathways can help in the early diagnosis and development of new clinical and pharmacological treatments for ASD.

This study investigated the relative performance of ten brinjal (Solamum melongena L.) varieties for yield in fall 2014 in Bahawalpur. The study was conducted at farm area of Islamia University of Bahawalpur. Ten brinjal varieties were evaluated for yield performance in a research trial following randomized complete block design. Significant differences existed in the yield generated by tested varieties. Significantly more yield was recorded in Shamli and Eggplant deep black followed by Advanta 306, Sandhya F1, Black boy, Black nagina and Advanta 305 in descending order. Twinkle star and Kalash F1 generated significantly less yield while the significantly least yield was recorded for Xingchangjishi than all the tested varieties. Whitefly Bemesia tabaci (Homoptera: Aleyrodidae) and jassid Amrasca biguttula biguttula (Homoptera: Cicadellidae) were the major sucking insects attacking this crop. Populations of both pest insects were recorded significantly more on Xingchangjishi while least populations of these pests were recorded on Egg plant deep black and Sandhya F1. Correlation of insect populations with yield showed inverse relationships. These results are important regarding varietal performance for yield test conducted for ten brinjal varieties. Varieties i.e.,Eggplant deep black and Shamli with significantly more yields are recommended for cultivation in this area to get more brinjal yield.

At present, 1.5 million species of fungi are estimated. Among these less than 5% have been described. Many fungal species cause disease in plants. These diseases cause major economic and production losses in the agricultural industry worldwide. Monitoring plant health and detecting the pathogen early are essential to reduce the disease spread, and facilitate effective management practices. DNA-based methods now provide essential tools for accurate plant disease diagnosis. Recently, effective amplification platforms, probe development, various quantitative PCR, DNA barcoding and RNA-Seq-based next-generation sequencing have revolutionized the research in fungal detection field, and differentiation area. Although the molecular diagnostics techniques have grown extensively over the last couple of decades but still there is a long way to go in the development and application of molecular diagnostics to assist the plant disease diagnosticians. Finally, molecular diagnostic techniques used in plant disease diagnostic clinics need to be robust, reliable, inexpensive and easy to be used that they can compete with, and complement traditional techniques. Challenge now remains residue with the researchers to develop the practical techniques used for diagnostic setting. Examples of the recent advancement in the molecular techniques for diagnosing the fungi causing plant disease are discussed in the review. Abbreviations: PCR: polymerase chain reaction; LAMP: loop-mediated isothermal amplification; RCA: rolling circle amplification; NASBA: nucleic acid sequence-based amplification; ITS: internal transcribed spacer; RT: reverse transcriptase; FEB: Fusarium ear blight; qPCR: real-time PCR; SNPs: single nucleotide polymorphism; HRCA: hyper-branched RCA; FIP: forward inner primer; BIP: backward inner primer; COX I: cytochrome c oxidase I; MBs: molecular beacons.

The present work is definitely an approach towards attaining price competency of bio-diesel to petroleum diesel. The oils extracted from abundantly available waste of Zahidi, Basra and Khazravi date seeds were used to produce biodiesel using acid (HCl), base (KOH), immobilized enzyme (lipase), immobilized enzyme/acid (lipase/HCl) and immobilized enzyme/base (lipase/KOH) catalyzed processes. Mixed catalysis (immobilized enzyme + acid or immobilized enzyme + base) resulted in better yields in comparison to acid or base catalysis. The properties of biodiesel were evaluated by fuel standard tests and the results were compared with EN14214 and ASTM D6751 standards. Biodiesel produced from date seed oil was found to have a high cetane number (55–60.3), low iodine value (44–50) and good flash point (135–140 °C). Pour point of pure biodiesel produced from Khazravi and Zahidi was found to range from 2 to −2 °C. Biodiesel produced from Basra exhibited good pour point (−4.7 to −8.3 °C) in comparison to other varieties. The components present in biodiesel produced from various date varieties were determined by gas chromatographic-mass spectrometric analyses (GCMS). The fatty acid (%) detected in date seed biodiesel were oleic acid (33.4–47.4), lauric acid (19–28), palmitic acid (13.6–19.2), myristic acid (13.6–17.44) and linoleic acid (6.4–8.5). A special feature of date seed oil biodiesel was the presence of considerable amounts of low chain fatty acids.

Postharvest disease management is one of the key challenges in commercial mango supply chains. Comprehensive investigations were made regarding the impact of geographic locality on postharvest disease development and other quality parameters in 'Sindhri' and 'Samar Bahisht (S.B.) Chaunsa' mangoes under ambient (33±1°C; 55-60% RH) and low temperature storage/simulated shipping (12±1°C; 80- 85% RH) conditions (28 or 35 days storage for 'Sindhri' and 21 or 28 days for 'S.B. Chaunsa'). Physiologically mature (days from fruit set were 95-100 and 110-115 for 'Sindhri' and 'S.B Chaunsa', respectively) 'Sindhri' and 'S.B. Chaunsa' fruits were harvested from five geographic localities and subjected to ambient and simulated shipping conditions. Under ambient conditions, no disease incidence was observed till fruit eating stage in 'Sindhri'. However, in 'S.B. Chaunsa', significant variation in different localities was observed with respect to disease incidence. Maximum and at par disease was exhibited by the fruit collected from district Vehari and Khanewal in 'S.B. Chaunsa'. Under simulated shipping conditions, disease development varied significantly with respect to different locations and storage durations. In 'Sindhri', fruit of M. Garh, while, 'S.B. Chaunsa' fruit of districts R.Y. Khan, M. Garh and Khanewal showed higher disease incidence. Fruit peel colour development was significantly reduced as storage days increased. Fruit firmness, skin shriveling, fresh weight loss, dry matter, biochemical and organoleptic attributes also varied significantly among the fruit sourced from different orchards of different localities. Analysis of N contents in leaves and fruit peel revealed that N contents of leaf and peel were positively correlated with disease severity in mango. Botryodiplodia spp., Phomopsis mangiferae, Alternaria alternata, Colletotrichum gloeosporioides were the pathogens isolated from fruits of all locations; however, the prevalence frequency varied with the geographic localities. In conclusion, the production locality, cultivar and nutrition (nitrogen content of fruit peel) had significant effect on fruit quality out-turn at ripe stage in terms of disease development so area specific disease management system needs to be implemented for better quality at retail.

Natural pathogen pressure is an important factor that shapes the host immune defense mechanism. The current study primarily aimed to explore the molecular basis of the natural immune defense mechanism of a sporadic pest, Gryllus bimaculatus, during swarming by constructing cDNA libraries of the female mid-gut, male mid-gut, testes, and ovaries. The Illumina HiSeq platform generated an average of 7.9 G, 11.77 G, 10.07 G, and 10.07 G bases of outputs from the male mid-gut, female mid-gut, testes, and ovaries and libraries, respectively. The transcriptome of two-spotted field crickets was assembled into 233,172 UniGenes, which yielded approximately 163.58 million reads. On the other hand, there were 43,055 genes in common that were shared among all the biological samples. Gene Ontology analysis successfully annotated 492 immune-related genes, which comprised mainly Pattern Recognition Receptors (62 genes), Signal modulators (57 genes), Signal transduction (214 genes), Effectors (36 genes), and another immune-related 123 genes. In summary, the identified wide range of immune-related genes from G. bimaculatus indicates the existence of a sophisticated and specialized broad spectrum immune mechanism against invading pathogens, which provides, for the first time, insights into the molecular mechanism of disease resistance among two-spotted field crickets.

*Corresponding author: BrairaWahid, Genome Centre for Molecular Based Diagnostics and Research, Al-Sudais Plaza Abdalian Cooperative Society, Lahore, Pakistan. Tel: +92-4235219020, E-mail: brairawahid@gmail.com

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Plants are frequently exposed to limitations in oxygen availability during their lifetime. During evolution, they have developed a number of physiological and morphological adaptations to tolerate oxygen and other stress conditions. These include regulation of growth by gene expression and ATP generation. The regulation of nuclear genes after hypoxia and anoxia is well studied; however, the regulation of mitochondrial genes in response to oxygen stress has not been characterized to date. Therefore, we have established an Arabidopsis mitochondrial genome-specific microarray that accommodates probes for all mitochondrial DNA-encoded genes and conserved open reading frames. Our analysis showed an up-regulation of mitochondrial transcripts in Arabidopsis roots after 48 h of hypoxia. Since no significant difference was detected in the expression of mitochondrial RNA polymerases or the mitochondrial DNA content per cell, we propose a transcriptional mode of induction of mitochondrial gene expression under hypoxia.

N-acetylcysteine (NAC) is a well-known antidote for acetaminophen toxicity and is easily available over the counter. It has antioxidant and anti-inflammatory properties and an established tolerance and safety profile. Owing to its neuroprotective effects, its clinical use has recently expanded to include the treatment of different psychiatric and non-psychiatric disorders. Although a number of randomized controlled trials have documented the clinical evidence for NAC, there are no reviews that summarize the evidence. The present scoping review summarizes the study designs, the patient characteristics, the evidence and the limitations in randomized controlled trials designed to explore the efficacy of NAC for psychiatric conditions in the pediatric population.

Spodoptera litura is an important polyphagous insect pest and information on its host plant relationship aspects is important for developing IPM approaches. The present study evaluates the effect of five natural hosts (cabbage, maize, okra, alfalfa and sesbania) and an artificial diet on the biology of S. litura under laboratory conditions. It was observed that cabbage is the most suitable host- as the larval length was found maximum on cabbage (3.53 cm) followed by alfalfa (3.50 cm), while the least was on okra (2.79 cm). The life span and fecundity of female was also maximum with cabbage; and the least number of egg masses of 7.5 and 9.5 were observed on maize and okra, respectively. The total number of eggs laid was maximum with alfalfa (7200) followed by cabbage (6000) and artificial diet (5200); the least being with maize (1900) and sesbania (2000). These results reveal that all the biological parameters viz., egg, larva, pupa and adult of S. litura are influenced by the host plants.