Molecular identification of Hymenopteran insects collected by using Malaise traps from Hazarganji Chiltan National Park Quetta, Pakistan.

The order Hymenoptera holds great significance for humans, particularly in tropical and subtropical regions, due to its role as a pollinator of wild and cultivated flowering plants, parasites of destructive insects and honey producers. Despite this importance, limited attention has been given to the genetic diversity and molecular identification of Hymenopteran insects in most protected areas. This study provides insights into the first DNA barcode of Hymenopteran insects collected from Hazarganji Chiltan National Park (HCNP) and contributes to the global reference library of DNA barcodes. A total of 784 insect specimens were collected using Malaise traps, out of which 538 (68.62%) specimens were morphologically identified as Hymenopteran insects. The highest abundance of species of Hymenoptera (133/538, 24.72%) was observed during August and least in November (16/538, 2.97%). Genomic DNA extraction was performed individually from 90/538 (16.73%) morphologically identified specimens using the standard phenol-chloroform method, which were subjected separately to the PCR for their molecular confirmation via the amplification of cytochrome c oxidase subunit 1 (cox1) gene. The BLAST analyses of obtained sequences showed 91.64% to 100% identities with related sequences and clustered phylogenetically with their corresponding sequences that were reported from Australia, Bulgaria, Canada, Finland, Germany, India, Israel, and Pakistan. Additionally, total of 13 barcode index numbers (BINs) were assigned by Barcode of Life Data Systems (BOLD), out of which 12 were un-unique and one was unique (BOLD: AEU1239) which was assigned for Anthidium punctatum. This indicates the potential geographical variation of Hymenopteran population in HCNP. Further comprehensive studies are needed to molecularly confirm the existing insect species in HCNP and evaluate their impacts on the environment, both as beneficial (for example, pollination, honey producers and natural enemies) and detrimental (for example, venomous stings, crop damage, and pathogens transmission).

Mapping phenotypic performance and novel SNPs for cold tolerance in tomato (Solanum lycopersicum) genotypes through GWAS and population genetics.

The cold stress susceptibility of tomato (Solanum lycopersicum) curtails its cultivation, with significant impact in temperate regions and on cropping seasons. To unravel genomic regions responsible for cold stress resilience, a diverse set of fifty genotypes encompassing cultivated, wild species, and landraces were genotyped using genotyping-by-sequencing. Over two years and six trials employing both early and late sowing, these lines were evaluated. Illumina-based next-generation sequencing produced up to 3 million reads per sample from individually sequenced library pools. The Tassel pipeline yielded 10,802 variants, subsequently filtered to 3,854 SNPs for genome-wide association analysis (GWAS). Employing clustering methods (population structure) via TASSEL, SNPhylo, and Kinship matrix, the fifty genotypes clustered into four distinct gene pools. The GWAS for cold tolerance in tomato integrated key traits including yield. Using six independent phenotypic datasets representing various environments, the study identified 4,517 significant marker-trait associations for cold tolerance traits. Notably, pivotal variations (> 10%) in cold stress tolerance, particularly proline content, were linked to marker-trait associations. Additionally, 5,727 significant marker-trait associations for yield and yield-related traits were unveiled, shedding light on fruit yield and directly associated attributes. The investigation pinpointed 685 candidate genes across all examined traits, including 60 genes associated with biological processes within these genomic regions. Remarkably, 7 out of the 60 genes were directly linked to abiotic stress tolerance, functioning as stress-responsive genes either directly or indirectly. The identified genes, particularly those associated with stress response, could hold the key to enhancing cold tolerance and overall crop productivity in tomato cultivation.

Geographic distribution and genetic diversity of Newcastle disease virus in pigeons from Pakistan.

RESEARCH HIGHLIGHTS: Virulent NDV genotypes were repeatedly isolated from pigeons.Evidence of epidemiological links among viruses isolated from various locations.Distinct phylogenetic branches suggest separate, simultaneous evolution of NDVs.Study information could be helpful in the development of an effective vaccine.

Growth Optimization and Rearing of Mealworm (Tenebrio molitor L.) as a Sustainable Food Source.

As a sustainable food source for humans, mealworms (Tenebrio molitor) have a great deal of potential, due to the fact that they have a very favorable nutritional profile and a low environmental impact. For meal production, feed formulation and optimization are important. The mealworm Tenebrio molitor (Coleoptera: Tenebrionidae) is the most consumed insect in the world. Mealworms were given a variety of diets, including wheat bran as constant diet supplemented with different levels of Ospor (Bacillus clausii) at 0.002 g, 0.004 g, 0.006 g, and 0.008 g; imutec (Lacticaseibacillus rhamnosus) at 0.2 g. 0.4 g, 0.6 g, and 0.8 g; fungi (Calocybe indica) at 250 g, 500 g, and 750 g; yeast (Saccharomyces cerevisiae) at 50 g, 100 g, and 150 g; and wheat bran (standard diet) were examined in complete randomized design (CRD). Different parameters, i.e., the larval, pupal, and adult weight, size, life span, and nutritional profile of mealworm were studied. When compared with other insect growth promoters, only wheat bran was discovered to be the most efficient. It generated the heaviest and longest larvae at 65.03 mg and 18.32 mm, respectively, as well as pupae weighing 107.55 mg and 19.94 mm, respectively, and adults weighing 87.52 mg and 20.26 mm, respectively. It was also determined that fungi (C. indica) and ospor (B. clausii) promoted faster larval development than yeast (S. cerevisiae) and imutec (L. rhamnosus). Larval mortality was also greater in the imutec (L. rhamnosus) and yeast (S. cerevisiae) diets than the others. No pupal mortality was recorded in all diets. Furthermore, the protein content of Tenebrio. molitor raised on a diet including fungi (C. indica) was the highest at (375 g), with a content of 68.31%, followed by a concentration of (250 g) with a content of 67.84%, and wheat bran (1 kg) (normal diet) with the lowest content at 58.91%. T. molitor larvae fed a diet supplemented with bacterial and fungal had lower fat and ash content than bran-fed T. molitor larvae (standard diet). Wheat bran (normal diet) had the highest fat at 16.11%, and ash at 7.71%. Hence, it is concluded that wheat bran alone or diet containing fungi (C. indica) and ospor (B. clausii) performed better in terms of growth, and these diets and protein content are recommended for the mass rearing of mealworms.

Multimodal Intervention Assessing the Appropriateness of Acid Suppression Therapy is Associated With Reduced Prescriptions at the Time of Discharge for Hospitalized Inpatients.

Background: Acid suppression therapy (AST), including proton pump inhibitors and histamine 2 receptor antagonists, are an overused class of medications. When used inappropriately, AST leads to polypharmacy, increased healthcare costs, and possible negative health consequences. Objective: To assess whether an intervention including prescriber education combined with a pharmacist-driven protocol was effective in reducing the percentage of patients who were discharged with inappropriate AST. Methods: This was a prospective pre-post study of adult patients who were prescribed AST before or during their admission to an internal medicine teaching service. All internal medicine resident physicians received education on appropriate AST prescribing. During the 4-week intervention period, dedicated pharmacists assessed the appropriateness of AST and made recommendations regarding deprescribing if no appropriate indication was identified. Results: During the study period, there were 14 166 admissions during which patients were prescribed AST. Out of the 1143 admissions during the intervention period, appropriateness of AST was assessed by a pharmacist for 163 patients. AST was determined to be inappropriate for 52.8% (n = 86) of patients and discontinuation or de-escalate of therapy occurred in 79.1% (n = 68) of these cases. The percentage of patients discharged on AST decreased from 42.5% before the intervention to 39.9% after the intervention (P = .007). Conclusion: This study suggests that a multimodal deprescribing intervention reduced prescriptions for AST without an appropriate indication at the time of discharge. To increase the efficiency of the pharmacist assessment several workflow improvements were identified. Further study is necessary to understand the long-term outcomes of this intervention.

Natural oxidation of Ti3C2Tx to construct efficient TiO2/Ti3C2Tx photoactive heterojunctions for advanced photoelectrochemical biosensing of folate-expressing cancer cells.

The rapid-charge carrier recombination and low conductivity are critical in devising an efficient photoelectrochemical (PEC) sensor. Herein, we propose partial oxidation of few-layered MXene (Ti3C2Tx) to construct a photo-active TiO2/Ti3C2Tx platform that could be configured for PEC sensing of folate receptors (FR), particularly, FR-expressing breast cancer cells (MDA-MB-231). MXene-Ti3C2Tx dispersion was oxidized in natural-open air conditions, where continuous exposure for six (06) days allowed for homogeneous in-situ growth of TiO2 over MXenes nanosheets (MX-06). This exposure enabled partial oxidation of MXene-sheets with a balanced TiO2 to MXene content that could exhibit improved photoresponsive characteristics owing to the synergism of redox-active TiO2 and highly conductive underlying Ti3C2Tx. The photoelectrode was then adapted for biorecognition by conjugating chitosan and folic acid (FA) networks, which permitted selective detection of FR-expressed cells with significant antifouling capabilities against common proteins such as bovine serum album (BSA), hemoglobin, and immunoglobulin G. (Ig G). The detection mechanism relies on FA's strong affinity for cancer cell folate receptors, which proportionally inhibited the photoelectrodes PEC oxidation response to ascorbic acid (AA)(mediator). The proposed inhibition strategy enabled sensitive detection of FR-expressed MDA-MB-231 cells in the concentration range of 1 × 102 to 2 × 107 cells/mL with a detection limit of 1.01 cells/mL (S/N = 3).

An Updated Review on Efficiency of Penthorum chinense Pursh in Traditional Uses, Toxicology, and Clinical Trials.

Traditional Chinese medicines (TCM) play an important role in the control and treatment of several animal diseases. Penthorum chinense Pursh (PCP) is a famous plant for its use in traditional medication practice and therapeutic effects in numerous pathological conditions. In China, PCP is utilized for both food and medication due to numerous bioactivities. PCP is widely administered in prevention and treatment of traumatic injury, edema, and liver diseases with functions of reducing swelling, support diuresis, blood stasis, and mitigation symptoms of excessive alcohol intake. Recently, PCP highlighted for research trials in various fields including pharmacology, pharmacognosy, cosmeceuticals, nutraceuticals, and pharmaceuticals due to medicinal significance with less toxicity and an effective ethnomedicine in veterinary practice. PCP contains diverse important ingredients such as flavonoids, organic acids, coumarins, lignans, polyphenols, and sterols that are important bioactive constituents of PCP exerting the therapeutic benefits and organ-protecting effects. In veterinary, PCP extract, compound, and phytochemicals/biomolecules significantly reversed the liver and kidney injuries, via antioxidation, oxidative stress, apoptosis, mitochondrial signaling pathways, and related genes. PCP water extract and compounds also proved in animal and humans' clinical trial for their hepatoprotective, antiaging, nephroprotective, anti-inflammatory, antidiabetic, antibacterial, antiapoptotic, immune regulation, and antioxidative stress pathways. This updated review spotlighted the current information on efficiency and application of PCP by compiling and reviewing recent publications on animal research. In addition, this review discussed the toxicology, traditional use, comparative, and clinical application of PCP in veterinary practices to authenticate and find out new perspectives on the research and development of this herbal medicine.

Genomic and comparative clinico-pathological assessment of two Pakistani pigeon-derived newcastle disease virus sub-genotypes XXI.1.1 and XXI.1.2 isolated in 2017.

Pigeon paramyxovirus type-1 (PPMV-1) is an antigenic-variant of Newcastle disease virus (NDV) which is associated with infection in Columbidae family. In this study, we isolated two pigeon-derived strains pi/Pak/Lhr/SA_1/17 (designed as SA_1) and pi/Pak/Lhr/SA_2/17 (designed as SA_2) from diseased pigeons collected in Punjab province in 2017. We performed the whole genome, phylogenetic analysis and comparative clinico-pathological evaluation of two viruses in pigeons. Phylogenetic analysis based on fusion (F) gene and complete genome sequences showed that SA_1 belonged to sub-genotype XXI.1.1 and SA_2 clustered in sub-genotype XXI.1.2. SA_1 and SA_2 viruses contributed to morbidity and mortality in pigeons. Remarkably, although the two viruses resulted in comparatively similar pattern of pathogenesis and replication ability in various tissues of infected pigeons, SA_2 could cause more severe histopathological lesions and had comparatively high replication ability in pigeons than SA_1. Moreover, pigeons infected with SA_2 had higher shedding efficiency than that of pigeons infected with SA_1. Moreover, several aa substitutions in the major functional domains of the F and HN proteins might be contributed to the pathogenic differences between the two isolates in pigeons. Overall, these findings provide us with important insight into the epidemiology and evolution of PPMV-1 in Pakistan and laid the foundation for the further elucidation of the mechanism underlying the pathogenic difference of PPMV-1 in pigeons.

A unilateral FLAIR-hyperintense lesions in anti-MOG-associated encephalitis with seizures (FLAMES) case from a developing country: A case report.

Introduction: The new clinical and radiological entity of the myelin oligodendrocyte glycoprotein (MOG) antibody-associated disease (MOGAD) is known as the FLAMES syndrome. It will add to the literature and enhance the understanding of this disease. Case presentation: Our case presented a 25-year-old male patient with no known comorbid presented with a generalized sudden headache of moderate intensity for 10 days who came to our hospital. A right-sided upper motor neuron facial palsy was found on the examination. The diagnosis was further confirmed utilizing the MRI scan and the presence of MOG-IgG antibodies. The patient was started on intravenous methylprednisolone which lead to improvement of his symptoms. In the follow-up contrast-enhancing MRI of the brain, the findings suggested near resolution as compared to the initial MRI. Discussion: The FLAMES have been categorized as a subtype of the MOGAD. It is commonly found in males as compared to females and the mean age in which it commonly affects is 29 years with the common age group of 11-46 years. The clinical symptoms and physical signs along with the findings of unilateral cortical FLAMES on MRI suggest the distinctive findings of the FLAMES syndrome. Patients commonly present with symptoms like fever, headache, and cortical symptoms like aphasia. Conclusion: There is a dire need of improving the understanding of this clinic-radiographic syndrome which makes it critically important to ensure the timely diagnosis and prompt consideration of the required medications.

Ecotoxicity assessment for environmental risk and consideration for assessing the impact of silver nanoparticles on soil earthworms.

Silver nanoparticles (AgNPs) are found in a range of commercial products due to their proven antibacterial properties. The unused silver nanoparticles (AgNPs) may make its way into the soil via biosolids that come from wastewater treatment or the effluent that comes from industrialisation processes, where it could be harmful to the organism that live in terrestrial ecosystems. In addition, silver ions are one of the most toxic forms of heavy metal released from dissolved silver nitrate (AgNO3) and AgNPs through dissolution or oxidation. The study examined the effect of engineered AgNPs, and AgNO3 on earthworms which are one of the most important bioindicator for determining toxicity in soil environment. Epigeic earthworm, Eudrilus eugeniae was exposed to soils spiked with equivalent concentrations of AgNPs or AgNO3 at 0, 10, 100, and 200 mg kg-1 in soil for 56 days of experiments. The survival and growth rate was recorded at 7th, 14th, 21st, 28th days and accumulation of Ag in earthworm tissue at 14th and 28th days, antioxidant enzymes at 28th days and reproduction at 56th days of experiment. Further, a short-term exposure of AgNPs and AgNO3 was conducted to observe avoidance behaviour after 48 h of exposure. The result indicated that survivability was relatively low on exposure of AgNO3 (83.3%) than AgNPs (86.7%) in 200 mg kg-1 spiked soils, besides the growth was inhibited in both AgNPs (3.68%) and AgNO3 (3.25%) at 28th days. The uptake of Ag from AgNO3 in the earthworm tissue was slightly higher than uptake of Ag from AgNPs and it showed concentration-dependent inhibitory effects on reproduction. In AgNO3 spiked soil, a high level of the Malondialdehyde (MDA) based lipid peroxidation and increased activity of antioxidant enzyme catalase (CAT) was observed than AgNPs spiked soil. Similarly, glutathione (GSH), a cofactor for GPx and GST enzymes, was lower in AgNO3-spiked soil than in AgNPs-spiked soil. In terms of avoidance behaviour, there was no discernible difference between the distribution of earthworms in AgNPs and AgNO3 after 48 h. The study found E. eugeniae exhibits concentration-dependent alterations in its competence to survive, antioxidant enzymes, and reproduction. AgNO3 was found to be more sensitive than AgNPs in the study. The research investigates the effect of AgNPs on earthworms in the soil ecosystem since this understanding is crucial for a comprehensive evaluation of AgNPs' environmental consequences.

Physicochemical and adulteration study of fresh milk collected from different locations in Pakistan.

The present research was carried out to assess raw milk's quality as collected from the commercial markets of the Mardan district (Khyber Pakhtunkhwa). The locality from which milk samples were collected included; Bijligar (BG), Manga (M), Chamthara (CM), Main Bazar Mardan (MB), Mahidherai (MD), and Sharif Abad (SB), located in district Mardan, Khyber Pakhtunkhwa (KP). A total of 36 milk samples were collected at the rate of 6 samples per location. The outcome of the data exhibited that the percentage of protein content was highest (3.34%) in MB and SA (3.30%), while lower percentages were recorded in sample M (3.03%) and CM 93.06%). Maximum pH were shown in M and BG to be 7.55 and 7.33, respectively. For fats content, the highest percentage of fats was witnessed in MB as 4.04%, and minimum fats content was noted in Sample M as 3.57%. Water content was highest in Sample SA and BG at 15.85% and 15.64%, respectively. Qualitative analysis of adulterants like detergents, Formalin, starch, and Hydrogen peroxide was also carried out for all the collected samples. Adulteration results were positive for all the milk samples, with the highest being in samples MB (30%), while all the remaining samples had adulteration at 20% each. Both MB and CM samples were adulterated with urea, while the remaining 4 were adulterated with neutralizers. Thus, it may be summarized from the whole analysis that the milk available in commercial markets of district Mardan was adulterated with different adulterants and is not recommended for consumption.

The SARS-CoV-2 differential genomic adaptation in response to varying UVindex reveals potential genomic resources for better COVID-19 diagnosis and prevention.

Coronavirus disease 2019 (COVID-19) has been a pandemic disease reported in almost every country and causes life-threatening, severe respiratory symptoms. Recent studies showed that various environmental selection pressures challenge the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infectivity and, in response, the virus engenders new mutations, leading to the emergence of more virulent strains of WHO concern. Advance prediction of the forthcoming virulent SARS-CoV-2 strains in response to the principal environmental selection pressures like temperature and solar UV radiation is indispensable to overcome COVID-19. To discover the UV-solar radiation-driven genomic adaption of SARS-CoV-2, a curated dataset of 2,500 full-grade genomes from five different UVindex regions (25 countries) was subjected to in-depth downstream genome-wide analysis. The recurrent variants that best respond to UV-solar radiations were extracted and extensively annotated to determine their possible effects and impacts on gene functions. This study revealed 515 recurrent single nucleotide variants (rcntSNVs) as SARS-CoV-2 genomic responses to UV-solar radiation, of which 380 were found to be distinct. For all discovered rcntSNVs, 596 functional effects (rcntEffs) were detected, containing 290 missense, 194 synonymous, 81 regulatory, and 31 in the intergenic region. The highest counts of missense rcntSNVs in spike (27) and nucleocapsid (26) genes explain the SARS-CoV-2 genomic adjustment to escape immunity and prevent UV-induced DNA damage, respectively. Among all, the most commonly observed rcntEffs were four missenses (RdRp-Pro327Leu, N-Arg203Lys, N-Gly204Arg, and Spike-Asp614Gly) and one synonymous (ORF1ab-Phe924Phe) functional effects. The highest number of rcntSNVs found distinct and were uniquely attributed to the specific UVindex regions, proposing solar-UV radiation as one of the driving forces for SARS-CoV-2 differential genomic adaptation. The phylogenetic relationship indicated the high UVindex region populating SARS-CoV-2 as the recent progenitor of all included samples. Altogether, these results provide baseline genomic data that may need to be included for preparing UVindex region-specific future diagnostic and vaccine formulations.

Comparative transcriptomics unravels new genes imparting scab resistance in apple (Malus x domestica Borkh.).

Apple scab is caused by an ascomycete fungus, Venturia inaequalis (Cke.) Wint., which is one of the most severe disease of apple (Malus × Domestica Borkh.) worldwide. The disease results in 30-40% fruit loss annually and even complete loss in some places. Owing to the evolving susceptibility of resistant apple genotypes harboring R-genes to new variants of V. inaequalis, a comparative transcriptome analysis using Illumina (HiSeq) platform of three scab-resistant (Florina, Prima, and White Dotted Red) and three susceptible (Ambri, Vista Bella, and Red Delicious) apple genotypes was carried out to mine new scab resistance genes. The study led to the identification of 822 differentially expressed genes in the tested scab-resistant and scab-susceptible apple genotypes. The most upregulated genes uniformly expressed in resistant varieties compared to susceptible ones were those coding for 17.3 kDa class II heat shock protein-like, chaperone protein ClpB1, glutathione S-transferase L3-like protein, B3 domain-containing protein At3g18960-like, transcription factor bHLH7, zinc finger MYM-type protein 1-like, and nine uncharacterized proteins, besides three lncRNAs. The genes that were downregulated in susceptible and upregulated in resistant cultivars were those coding for non-specific lipid transfer protein GPI-anchored 1, rust resistance kinase Lr10-like, disease resistance protein RPS6-like, and many uncharacterized proteins. DESeq2 analysis too revealed 20 DEGs that were upregulated in scab-resistant cultivars. Furthermore, a total of 361 genes were significantly upregulated in scab-susceptible variety, while 461 were found downregulated (P value < 0.05 and Log2 (FC) > 1). The differentially expressed genes (DEGs) were related to various pathways, i.e., metabolic, protein processing, biosynthesis of secondary metabolites, plant hormone signal transduction, autophagy, ubiquitin-mediated proteolysis, plant-pathogen interaction, lipid metabolism, and protein modification pathways. Real-time expression of a set of selected twelve DEGs further validated the results obtained from RNA-seq. Overall, these findings lay the foundation for investigating the genetic basis of apple scab resistance and defense pathways that might have a plausible role in governing scab resistance in apple against V. inaequalis.

Surveillance and Assessment of Risk Factors for Newcastle Disease Virus from Live Bird Retail Stalls in Lahore District of Pakistan.

Live bird markets (LBMs) in Asian countries are considered hubs for the spread of several poultry viruses. In Pakistan, there is a lack of uniformity in practices used in LBMs, which leads to the spread of poultry diseases. A cross-sectional survey was conducted in June-October 2017 to determine the circulation of Newcastle disease virus (NDV) in chickens being sold in live bird retail stalls (LBRSs) and to identify potential risk factors associated with estimated prevalence. A total of 189 stalls (n = 1134 birds) distributed in eight administrative towns of Lahore were visited. A pool of six oropharyngeal swabs was collected from each stall and tested by real-time reverse transcriptase PCR for the presence of NDV. Forty-two out of 189 swabs were found positive with an overall prevalence of 22.22% (95% confidence interval [Cl]: 16.88%-28.67%). Data for 11 potential risk factors acquired through questionnaires were analyzed by survey-weighted logistic regression and prevalence odds ratios (ORs) for associated risk factors were calculated. A final multivariable model identified three risk factors for NDV prevalence in LBRSs, including trading other poultry breeds alongside broilers (OR = 2.41; 95% confidence interval [CI] = 1.5-6.1), purchasing birds from mixed sources (OR = 3.12; 95% CI = 1.4-11.9), and number of birds sold per day (OR = 6.32; 95% CI = 1.9-23.5). Additionally, 24 selected samples were sequenced and phylogenetic analysis of the complete fusion gene (1662 bp) revealed that all isolates belonged to Subgenotype VII.2. This study provides important information on the epidemiology of NDV in Pakistan and highlights the importance of implementing surveillance and biosecurity practices in LBRSs.

Vigilancia y evaluación de factores de riesgo para el virus de la enfermedad de Newcastle en puestos de venta al menudeo de aves vivas en el distrito de Lahore en Pakistán. Los mercados de aves vivas (LBM, por sus siglas en inglés) en los países asiáticos se consideran centros de propagación de varios virus aviares. En Pakistán, existe una falta de uniformidad en las prácticas utilizadas en los mercados de aves vivas, lo que conduce a la propagación de enfermedades avícolas. Se realizó una encuesta transversal de junio a octubre del 2017 para determinar la circulación del virus de la enfermedad de Newcastle (NDV) en pollos que se venden en puestos minoristas de aves vivas y para identificar posibles factores de riesgo asociados con la prevalencia estimada. Se visitó un total de 189 puestos (n = 1134 aves) distribuidos en ocho ciudades administrativas de Lahore. Se recolectó un grupo de seis hisopos orofaríngeos de cada puesto y se analizó mediante transcripción reversa y PCR en tiempo real para detectar la presencia del virus de Newcastle. Cuarenta y dos de los 189 hisopos resultaron positivos con una prevalencia general del 22.22 % (intervalo de confianza [IC] del 95 % = 16.88­28.67). Los datos para 11 factores de riesgo potenciales adquiridos a través de cuestionarios se analizaron mediante regresión logística ponderada por encuesta y se calcularon las razones de probabilidad (OR) de prevalencia para los factores de riesgo asociados. Un modelo multivariable final identificó tres factores de riesgo para la prevalencia del virus de Newcastle en puestos minoristas de aves vivas, incluido el comercio de otras razas de aves de corral junto con pollos de engorde (OR = 2.41; IC del 95 % = 1.5­6.1), la compra de aves de fuentes mixtas (OR = 3.12; IC del 95 % = 1.4 ­11.9), y número de aves vendidas por día (OR = 6.32; IC 95% = 1.9­23.5). Además, se secuenciaron 24 muestras seleccionadas y el análisis filogenético del gene de fusión completo (1662 pb) reveló que todos los aislamientos pertenecían al subgenotipo VII.2. Este estudio brinda información importante sobre la epidemiología del virus de Newcastle en Pakistán y destaca la importancia de implementar prácticas de vigilancia y bioseguridad en los en puestos minoristas de aves vivas.

Investigating the role of potassium and urea to control fruit drop and to improve fruit quality of "Dhakki" date palm.

Fruit drop is a key issue with date palm cultivars that can be addressed with a variety of methods and strategies. Foliar application of macronutrients can be more effective in inhibiting fruit drop and improving the quality of date fruits. The current study was carried out to investigate the possible role of potassium (K) and urea to reduce fruit drop and improve the fruit quality of "Dhakki" date palm. It was conducted in a complete randomised block design with seven treatments and three replications at Pakistan's Agricultural Research Institute, Dera Ismail Khan. The treatments used were: (i) Control (distilled water spray); (ii) Potassium sulphate (K2SO4) at 1 %; (iii) K2SO4 at 1 % + Urea at 2 %; (iv) K2SO4 at 2 %; (v) K2SO4 at 2 % + Urea at 2 %; (vi) K2SO4 at 3 % and; (vii) K2SO4 at 3 % + Urea at 2 %. All the concentrations were sprayed at Kimri stage of fruit development during two consecutive growing seasons. Twenty-one date palms of equal size and age were chosen for the assessments to measure percent fruit drop and other physicochemical variables, including fruit length, fruit diameter, fruit weight, pulp percentage, yield/bundle, pH, total soluble solids (TSS), K content in fruit, and all sugars (percent) of harvested date fruit. The results revealed that bunch spray of K significantly affected all the parameters during both seasons. Application of K2SO4 alone and in combination with urea not only effectively reduced the fruit drop but also improved fruit quality in date where, K2SO4 applied at 2 % combined with urea was the best concentration in reducing fruit drop, enhancing other physicochemical attributes, and improving fruit quality of "Dhakki" date palm. This study may effectively contribute to reduce the fruit drop and enhance the fruit quality by using K and urea, enabling farmers to improve the date yield and increase economic growth.

A DNA barcode survey of insect biodiversity in Pakistan.

Although Pakistan has rich biodiversity, many groups are poorly known, particularly insects. To address this gap, we employed DNA barcoding to survey its insect diversity. Specimens obtained through diverse collecting methods at 1,858 sites across Pakistan from 2010-2019 were examined for sequence variation in the 658 bp barcode region of the cytochrome c oxidase 1 (COI) gene. Sequences from nearly 49,000 specimens were assigned to 6,590 Barcode Index Numbers (BINs), a proxy for species, and most (88%) also possessed a representative image on the Barcode of Life Data System (BOLD). By coupling morphological inspections with barcode matches on BOLD, every BIN was assigned to an order (19) and most (99.8%) were placed to a family (362). However, just 40% of the BINs were assigned to a genus (1,375) and 21% to a species (1,364). Five orders (Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera) accounted for 92% of the specimens and BINs. More than half of the BINs (59%) are so far only known from Pakistan, but others have also been reported from Bangladesh (13%), India (12%), and China (8%). Representing the first DNA barcode survey of the insect fauna in any South Asian country, this study provides the foundation for a complete inventory of the insect fauna in Pakistan while also contributing to the global DNA barcode reference library.

Preference and performance of peach fruit fly (Bactrocera Zonata) and Melon fruit fly (Bactrocera Cucurbitae) under laboratory conditions.

In Pakistan and all over the world, the Peach Fruit Fly (PFF), Bactrocera zonata (Saunders.) and the Melon Fruit Fly (MFF), Bactrocera cucurbitae (Coquillett.) are considered severe and polyphagous insect pests for various fruits and vegetables. The current study was conducted to check the Laboratory preference and performance of B. cucurbitae and B. zonata on selected Fruits Citrus (Citrus sinensis), Apple (Malus domestica), Banana (Musa acuminate), and vegetable, Sponge gourd (luffa aegyptiaca), Bitter gourd (Momordica charantia) Pumpkin (Cucurbita moschata) under laboratory conditions. The study showed that Sponge Gourd was the preferable host with the mean pupae resurgence of (242.33), followed by Bitter Gourd (78.333) among selected vegetables. At the same time, among fruits, a banana was the preferable host with mean pupae resurgence (204.33), followed by orange (158.33). The pumpkin and apple was the least preferable host for both B. cucurbitae and B. zonata, with mean pupae resurgence (35.667) and (79.000), respectively. Furthermore, the study showed that Banana was the preferable host for B. Zonata among intact and infested fruits, whereas B. cucurbitaee showed the most preference to Bitter gourd among intact and infested vegetables showing significantly different results among intact and infested fruits and vegetables. Maximum number of eggs, pupa, female flies, male flies, adult emergence from pupa (flies) and period of pupa of B. zonata and B. cucurbitae on banana and bitter gourd. While, other fruits and vegetables showed the minimum number of eggs, pupa, female flies, male flies, adult emergence from pupa (flies) and period of the pupa. The current study concluded there is a need to evaluate other host plants against these fruit fly species for effective control.

Ni Nanoparticles Embedded Ti3C2Tx-MXene Nanoarchitectures for Electrochemical Sensing of Methylmalonic Acid.

MXenes-Ti3C2Tx, based on their versatile surface characteristics, has rapidly advanced as an interactive substrate to develop electrochemical sensors for clinical applications. Herein, Ni embedded Ti3C2Tx (MX-Ni) composites were prepared using a self-assembly approach where Ti3C2Tx sheets served as an interactive conductive substrate as well as a protective layer to nickel nanoparticles (Ni NPs), preventing their surface oxidation and aggregation. The composite displayed a cluster-like morphology with an intimate interfacial arrangement between Ni, Ti3C2Tx and Ti3C2Tx-derived TiO2. The configuration of MX-Ni into an electrochemical sensor realized a robust cathodic reduction current against methylmalonic acid (MMA), a biomarker to vitamin B12 deficiency. The synergism of Ni NPs strong redox characteristics with conductive Ti3C2Tx enabled sensitive signal output in wide detection ranges of 0.001 to 0.003 µM and 0.0035 to 0.017 µM and a detection sensitivity down to 0.12 pM of MMA. Importantly, the sensor demonstrated high signal reproducibility and excellent operational capabilities for MMA in a complex biological matrix such as human urine samples.

Comparative conventional preventive strategies for insect pest of okra.

The use of natural substances for pest control in agriculture is economically viable. It benefits both the human being and the environment due to its low persistence and toxicity. Therefore, the biopesticidal potential of three- plants-derived extracts (clove [Syzygium aromaticum], Hing [Hing (Asafetida)], and Wood Ash [Eucalyptus globulas]) was evaluated against different ' 'insect's pests on five okra varieties. All the treatments were sprayed at two stages, 1st before flowering and 2nd at the fruit-bearing stage. The results of the 24 h pre-spray revealed that the mean density of Aphis gossypii, Erias insulana, and Bemisia tabaci were significantly lower on a Shehzadi variety. However, among the treatments mean density of the A. gossypii and E. insulana after 1st and 2nd treatments were substantially more bass with E. globulas. Moreover, the Mean density of aphids was significantly lower after 72 h and 1-week time intervals. Furthermore, after 1st and 2nd treatments, the B. tabaci was considerably lower with hing on Shehzadi variety. It was found in the present study that the yield of five okra varieties was affected significantly by the application of the three treatments-pesticides. Among the various treatments, the application with E. globulas recorded a considerably higher crop yield. Therefore, clove, hing and E. globulas could be effective as alternative pest management methods. Furthermore, biopesticides generally are encouraged since they can proffer the solution of controlling insect pests without any environmental concern.

Identification of the economics, composition, and supplementation of maggot meal in broiler production.

The primary goal of this study was to conduct a preliminary assessment of maggot meal protein supplements in broiler production. Maggot meal comprises 55 percent crude protein (CP), 27.65% ether extract, 8.33% Ash, 3.37 crude fiber (CF), 2.14 NFE, 94.7 percent Dry Matter, 5702 kcal/kg gross energy, and 3955 kcal/kg metabolizing energy, according to proximate analysis. Maggot meal supplementation affected broiler meat feed intake, body weight gain, FCR, dressing %, mortality, antibody titer against ND, and organoleptic features. The cost-effectiveness of maggot supplementation was also evaluated. Birds were put into four experimental groups after a week of adaptation:Control group (M0), the first experimental group (M1), the second experimental group (M2), and the third experimental group (M3), which received supplements of 0, 2, 3, and 4 g/kg, respectively. A plane ratio was given to the control group as well. The overall feed intake findings were inversely proportional to the supplementation rate. Thus, the highly supplemented group (M3) showed the lowest feed intake than the control group (M0). Bodyweight gain was directly proportional to the supplementation rate, as evident by a considerable increase in the highly supplemented group (M3) compared to the control group.