Oryctes rhinoceros (coconut rhinoceros beetle) is a major pest of oil palm in Malaysia, with its impact intensified under zero-burn replanting systems that create persistent breeding sites. The emergence of the CRB-G (Guam) haplotype, which exhibits reduced susceptibility to Oryctes rhinoceros nudivirus (OrNV), has further compromised conventional biological control strategies, highlighting the need for updated management approaches. This study presents a PRISMA-compliant systematic review, complemented by exploratory bibliometric analysis, to synthesize evidence on the biology, establishment, and management of O. rhinoceros in Malaysia. Literature published between 1970 and 2025 was retrieved from major scientific databases using predefined PICOS criteria, yielding 14 studies eligible for qualitative synthesis. The review indicates that residue-intensive replanting practices significantly enhance beetle population persistence, whereas biological control measures, particularly Metarhizium anisopliae-based formulations, demonstrate more consistent field efficacy against CRB-G populations than OrNV alone. Pheromone trapping remains an effective tool for monitoring and population suppression when integrated within comprehensive integrated pest management (IPM) programs, while chemical control is increasingly limited by environmental and sustainability concerns. Bibliometric trends reveal a progressive research shift from chemical-based interventions toward biological, molecular, and sustainable IPM strategies. This review consolidates current knowledge, identifies critical gaps regarding CRB-G adaptation, and outlines priorities for developing resilient, sustainable management strategies in Malaysian oil palm systems.
Bacterial blight of walnut (Juglans regia L.), caused by the Gram-negative bacterium Xanthomonas arboricola pv. juglandis, has recently emerged as a serious threat to walnut production in the temperate regions of Azad Jammu and Kashmir (AJK), Pakistan. The present study provides the first comprehensive surveillance and biochemical characterization of walnut bacterial blight across major walnut-producing districts of AJK. Systematic field surveys were conducted in eight districts, with ten locations surveyed per district, to assess disease prevalence, incidence, and severity on walnut leaves and fruits using a standardized 0-4 disease rating scale. The mean percent disease index (PDI) across the region ranged from 69.00% to 76.63%, indicating moderate to very high disease pressure. Disease prevalence varied between 60% and 100%, reflecting widespread distribution of the disease, while disease incidence reached 100% at several locations. The pathogen was consistently isolated from symptomatic tissues and produced characteristic yellow, mucoid colonies on nutrient agar and yeast dextrose calcium carbonate media. Biochemical characterization revealed typical reactions of Xanthomonas spp., including a Gram-negative reaction, positive catalase activity, and a negative oxidase test. Pathogenicity assays conducted on walnut leaflets and immature fruits successfully reproduced typical blight symptoms, and Koch’s postulates were fulfilled through re-isolation of the pathogen. Overall, the findings demonstrate severe disease pressure in the region and confirm X. arboricola pv. juglandis as the primary causal agent of walnut bacterial blight in AJK. This study provides a critical baseline for future epidemiological investigations and supports the development of integrated, region-specific disease management strategies.
This study investigated the impact of heat stress on reproductive development in two pepper varieties, Cayenne and Yolo Wonder, by comparing two planting dates (March 20th and May 1st) in Iraq. The experimental design was a Randomized Complete Block Design in a 2×2 factorial arrangements. The later planting date exposed plants to severe heat stress, which significantly impaired all measured reproductive parameters. A dramatic increase in flower abscission was observed, particularly in Yolo Wonder (67.85%), along with a severe reduction in fruit set percentage (54.9% for Cayenne vs. 20.8% for Yolo Wonder). Heat stress also negatively affected fruit morphology, reducing average fruit weight to 51.8g for Cayenne and 74.8g for Yolo Wonder. Seed quality was considerably compromised, as an evident reduction in the number of seeds per fruit (95.0 for Cayenne vs. 62.4 for Yolo Wonder) and final germination percentage (88.36% for Cayenne vs. 65.05% for Yolo Wonder). Statistical analysis confirmed significant main effects of variety and planting date, as well as a significant interaction for most traits, underscoring the differential response of the two varieties. The Cayenne variety consistently exhibited superior thermotolerance across all parameters, highlighting its greater genetic resilience compared to the highly sensitive Yolo Wonder. Therefore, it is recommended to cultivate the heat-sensitive Yolo Wonder variety only during spring or autumn to avoid summer heat. In contrast, the heat-tolerant Cayenne variety can be successfully cultivated during summer months, maintaining acceptable productivity under heat stress conditions. These findings emphasize the critical importance of variety selection and planting schedule optimization for sustainable pepper production in heat-prone regions like Iraq.
Botanical insecticides are a sustainable option to synthetic compounds in controlling insect pests of stored grains and products. Datura innoxia contains bioactive alkaloids with established insecticidal and behavioral activity, however, its activity, against the southern cowpea beetle, Callosobruchus maculatus, is poorly studied. This research was carried out to explore the alkaloid profiles of alcoholic extracts of D. innoxia leaves and seeds on reproductive performance of C. maculatus as well as behavioral responses. The alkaloid richness of the plant was confirmed by the presence of two major tropane alkaloids in the leaf extract, hyoscyamine (5.1 ppm; retention time 3.68 min) and scopolamine (7.8 ppm; retention time 0.6 04 min) as revealed by high performance liquid chromatography analysis. The treatment of cowpea seeds using alcoholic leaf extracts caused a significant reduction in oviposition of C. maculatus females. Seed extracts performed better than leaf extracts. Treatments did not show any major variations among concentrations, which led to a high process of activity even in case of low doses. The assays conducted with repellency showed concentration-based effects, the highest repellency rates of 80 and 90 percent respectively were observed when leaf and seed extracts were used at a 3000 ppm concentration. The highly decreased reproduction and the effective repellencies can be linked to the toxicity of alkaloids, oviposition and disruption in insect-development. In general, the results demonstrate the potential of D. innoxia, especially seed extracts, as a green and efficient botanical protectant to preserve cowpea seeds.
Matiari district, located in Sindh Province, Pakistan, is endowed with diverse agricultural resources, including rice paddies, guava orchards, and crops such as barley, wheat, and lucerne (Medicago sativa). Beetle populations were surveyed in Matiari and neighboring areas, Hala, Saeedabad, Sekhat, Banoth, Odero Lal, and Bhitshah, during 2022-2023 using light traps and pitfall traps to collect adult specimens. Beetles (order Coleoptera) represent one of the most diverse insect orders globally, with over 400,000 described species. A total of 709 beetle specimens were collected, of which 10 species, representing four families, were accurately identified. The recorded species included Anthia sexguttata (Fabricius, 1775), Julodis euphratica (Laporte and Gory, 1835), Melolontha furcicauda (Ancey, 1881), Chiloloba acuta (Wiedemann, 1823), Protaetia indica (Mikšić, 1965), Carabus cashmirensis (Redtenbacher, 1844), Chlaenius hamifer (Chaudoir, 1856), Chlaenius quadricolor (Oliver, 1790), Catharsius pithecius (Fabricius, 1775), and Hycleus lugens (Latreille, 1817). Among these, Catharsius pithecius was the most abundant species. Most beetles were collected from muddy soils and cultivated fields during July, August, and September, coinciding with peak crop growth. Members of the family Carabidae are recognized as important biological control agents and bioindicators of environmental health. Scarabaeidae play key roles in nutrient cycling, pest regulation, and seed dispersal. Meloidae species are primarily agricultural pests, whereas Buprestidae include species that damage forestry and agricultural systems, although some contribute to decomposition and nutrient cycling. Despite their ecological significance, comprehensive studies on beetle diversity in Pakistan remain scarce.
This study investigates the fitness costs and biochemical implications associated with spinetoram resistance in Earias vittella (Lepidoptera: Noctuidae), a major cotton pest in Pakistan. Three experimental approaches were employed: (i) evaluation of fitness costs using life table parameters, (ii) assessment of insecticide-synergist interactions, and (iii) analysis of biochemical changes in resistant populations. Spinetoram-selected populations (SPINE-SEL) were compared with an unselected susceptible population (UNSEL). The results revealed significant fitness costs in the resistant populations. Developmental durations were markedly prolonged, with total development time increasing from 23.2 days in the unselected population to 30.4 days in the resistant population. Fecundity was significantly reduced, with susceptible females laying an average of 295.6 eggs compared to 197.4 eggs in resistant females. Mating success was also lower in resistant populations (56%) than in the unselected controls (82%). Biochemical assays indicated significantly elevated activities of detoxification enzymes, including mixed-function oxidases (MFOs), glutathione S-transferases (GSTs), and general esterases, in the resistant populations. Synergist bioassays revealed a 6.8-fold increase in spinetoram toxicity when combined with piperonyl butoxide (PBO) against resistant strains, highlighting the potential of synergists to mitigate resistance. Overall, these findings demonstrate substantial metabolic and reproductive trade-offs associated with spinetoram resistance and underscore the importance of integrated pest management strategies for the sustainable control of E. vittella in Pakistan.
Stem-end rot (SER) is a major postharvest disease that limits mango (Mangifera indica L.) production and export potential in Pakistan. This study developed a sustainable SER management framework by integrating pathogen characterization with chemical control strategies. Fungal isolates collected from six districts of Punjab were identified morphologically and molecularly as Lasiodiplodia theobromae, Colletotrichum gloeosporioides, Curvularia spicifera, and Alternaria alternata. Pathogenicity assays confirmed their role in SER, with L. theobromae producing the largest lesions (74.6 mm). In vitro fungicide screening revealed fludioxonil as the most effective compound, inhibiting mycelial growth by 84.6-94.4% across all pathogens at 500-2000 ppm. Field trials showed that fludioxonil at 2000 ppm reduced SER incidence and severity by 87.1% and 92.1%, respectively, compared to untreated controls (71.3% incidence). Azoxystrobin and difenoconazole demonstrated moderate efficacy, while fluoxastrobin was the least effective. Phylogenetic analysis indicated close genetic relatedness between Pakistani isolates and those from India and China, suggesting regional pathogen dispersal. These results establish fludioxonil as a highly effective fungicide for SER management under Pakistani conditions. To mitigate resistance development and enhance sustainability, its use should be integrated with cultural practices and biocontrol agents. Overall, this study provides a science-based, region-specific integrated disease management model to improve mango quality, reduce postharvest losses, and strengthen Pakistan’s export competitiveness.
Chilgoza pine (Pinus gerardiana) forests in Upper Dir are facing serious threats due to poor natural regeneration and slow growth. This study was conducted to isolate and identify plant growth-promoting rhizobacteria (PGPR) from the rhizospheric soils and root saplings of Chilgoza pine, evaluate their effects on seed germination and seedling growth, and compare their performance with phytohormones. The study area was Narkoon, located in the Pathrak forest range of Upper Dir district. The isolated bacterial strains were Enterobacter (from soil) and Klebsiella spp. (from roots). Chilgoza pine seeds were treated with gibberellic acid (150 ppm), PGPR, and their combinations. The experiment was carried out under controlled conditions in a growth chamber (CRD) and in a forest nursery (RCBD). Germination was recorded when the radicle reached 2 mm in length, and growth parameters were measured after one growing season. One-way ANOVA was applied to assess significant differences among treatments. The results showed that seeds treated with Klebsiella pneumoniae + 150 ppm gibberellic acid exhibited the highest germination energy (83%). Both Enterobacter cancerogenus and Klebsiella pneumoniae, alone or in combination with gibberellic acid, significantly enhanced germination percentage, mean daily germination, germination index, and seedling growth traits. Notably, (E. cancerogenus + 150 ppm gibberellic acid) resulted in the greatest root length (7.8 cm) and shoot length (14.6 cm). Statistical analysis confirmed significant treatment effects on germination and growth parameters (p 0.05). Overall, the study demonstrates that PGPR inoculation combined with gibberellic acid can substantially improve seed germination and seedling growth in P. gerardiana.
Leafhoppers (Hemiptera: Cicadellidae) constitute one of the most diverse and economically significant groups of phytophagous insects; however, their diversity and distribution in the arid regions of Pakistan remain poorly documented. The present study provides a comprehensive taxonomic and distributional assessment of Cicadellidae from Tharparkar and adjacent areas of Sindh, Pakistan, based on extensive field surveys conducted during 2023-2024. Specimens were collected using sweep nets, yellow sticky traps, and light traps from four major localities, Umerkot, Nangarparkar, Viravah, and Kaloi, and were identified using detailed morphological characteristics, with particular emphasis on male genitalia. A total of 19 species belonging to 15 genera, 9 tribes, and 3 subfamilies (Deltocephalinae, Megophthalminae, and Cicadellinae) were recorded. Deltocephalinae was the dominant subfamily in terms of both species richness and abundance. Among the recorded species, Cofana unimaculata (Cicadellinae) and Aconurella prolixa (Deltocephalinae) were the most abundant, indicating strong adaptation to hot, arid conditions and grass-dominated agro-ecosystems. Nangarparkar exhibited the highest overall species abundance, likely due to comparatively better vegetation cover and moisture availability, whereas Kaloi showed the lowest diversity, reflecting poor soil fertility and limited host plant availability. This study represents the first comprehensive checklist of Cicadellidae from Tharparkar and provides baseline data essential for future biodiversity assessments, ecological studies, and the development of pest management strategies in the arid and semi-arid ecosystems of Pakistan.
Copper resistance in citrus orchards is an emerging concern, requiring effective strategies to manage the spread of bacterial citrus canker. During a two-year surveillance (2020-2021), 5,490 citrus canker lesion samples were collected from major citrus-growing regions of Pakistan, including Punjab, Khyber Pakhtunkhwa, and the Federal area. Of these, 3,160 isolates were successfully cultured, giving an overall recovery rate of 58%, with variation across regions, districts, and cultivation zones. Sargodha district, the primary citrus-producing area of Punjab, was the main focus for copper-resistance assessment. A total of 120 bacterial isolates, along with a copper-resistant reference strain (LH-201), were evaluated on nutrient glucose agar amended with copper sulfate at 0.5 g, 1 g, and 2 g concentrations. Phenotypic characterization showed that most isolates tolerated 0.5 g CuSO₄ (60% recovery), fewer survived at 1 g (24%), and none grew at 2 g. In contrast, the reference strain (LH-201) exhibited complete resistance at all tested levels. Copper-tolerance patterns also varied among Sargodha sub-locations: isolates from Chak 48 and Choranga (2020) and Chak 82 (2021) showed higher recovery at 1 g CuSO₄. Overall, resistance frequency was greater in 2020 than in 2021. Molecular screening using primers targeting copper-resistance genes (CopLAB) detected no such genes in any isolates, despite their phenotypic tolerance at lower copper concentrations. Overall, Pakistani citrus canker populations were largely copper-sensitive, particularly at higher concentrations, with no genetic evidence of stable copper-resistance mechanisms. These findings highlight spatial and temporal variation in pathogen recovery and provide important understandings for copper-based management practices in Pakistan.
Mushrooms are increasingly recognized as valuable sources of bioactive compounds with potential applications in agriculture and medicine. Among them, Morchella rufobrunnea has gained attention for its antimicrobial and antifungal properties. However, reports of its occurrence and biological activity remain limited in many regions, including Iraq. The present study aimed to identify M. rufobrunnea isolates collected from Iraq using molecular tools and to evaluate their antifungal potential against major phytopathogenic fungi. Molecular identification was performed by sequencing the internal transcribed spacer (ITS) region of DNA. PCR amplification yielded products of approximately 550 bp, and phylogenetic analysis confirmed the specimens as M. rufobrunnea. Sequence alignment revealed 99% similarity with reference sequences in the NCBI database, specifically matching accession number OQ808797.1. This represents the first confirmed report of M. rufobrunnea in Iraq, expanding its known geographical distribution. The antifungal activity of ethanolic extracts of M. rufobrunnea was evaluated against Rhizoctonia solani, Fusarium oxysporum, F. graminearum, Alternaria alternata, and Sclerotium rolfsii. A concentration-dependent inhibitory effect was observed, with inhibition indices ranging from 24.70% to 55.03%. The highest antifungal activity was recorded at 1000 mg/100 ml, where F. graminearum showed maximum susceptibility (55.03%), while R. solani exhibited the lowest inhibition (36.93%). Statistical analysis confirmed significant differences among treatments (LSD₀.₀₅ = 3.79-6.51). These findings highlight both the taxonomic significance and the biocontrol potential of M. rufobrunnea. Its strong antifungal activity suggests promising applications in sustainable plant disease management.
Azolla fern, rich in proteins (up to 30%), carbohydrates, and vitamins, holds nutritional, environmental, and economic importance as a biofertilizer, animal feed, and water purifier. Its occurrence in Iraq has been reported in Kalar, the Hawr al-Hartha marshes near Basra, and the Tigris River, with studies from 2015 to 2023 confirming its local distribution. In the present study, three Azolla samples were collected from different locations: Abu Ghraib district in Baghdad province, Karbala province, and Salah al-Din province. DNA was extracted and amplified to assess the genetic diversity of the available Azolla species. PCR analysis revealed that the ferns A. microphylla and A. nilotica each produced a single DNA band. Sequence analysis showed 99.2% similarity of one sample with A. microphylla (accession number PQ587101.1), thereby identifying it as A. microphylla. Another sample showed 99% similarity with A. nilotica (accession number PQ587100.1), confirming its identification as A. nilotica. A phylogenetic tree was constructed using the NCBI-BLASTn program. Comparative analysis of the nucleotide sequences with reference data from the National Center for Biotechnology Information (NCBI), using BioEdit software, indicated that the A. nilotica isolate shared 99% similarity with an American isolate, while A. microphylla showed 99% similarity with Indian isolates. This study provides new insights into the biodiversity of Azolla in Iraq and identifies its species through molecular characterization.
Apis mellifera L. face growing threats from sub-lethal pesticide exposure that disrupts their physiology and behavior, endangering colony survival and pollination efficiency. The present study evaluated the residual sub-lethal effects of bifenthrin on key health biomarkers, social interactions, and foraging behavior of worker honey bees under laboratory and semi-field conditions. Bees were exposed to four bifenthrin treatment levels along with a control group. Phenoloxidase (PO) activity, lifespan, antennation, trophallaxis, and foraging behavior were assessed using standard protocols and subjected to statistical analysis. PO activity was highest in the control group (26.64 ± 1.11 U/mg tissue) and lowest in the T1 treatment (8.90 ± 0.59 U/mg tissue). Lifespan declined from 43.00 ± 0.58 days (control) to 34.33 ± 0.33 days (T1). Similarly, antennation and trophallaxis frequencies decreased from 65.00 ± 4.93 and 45.00 ± 4.04 bees/event (control) to 37.00 ± 3.46 and 21.33 ± 3.18 bees/event (T1), respectively. Foraging activity, including incoming and pollen-carrying bees, also declined significantly, from 349.00 ± 17.58 to 237.00 ± 11.59 bees/5 min and from 121.67 ± 6.12 to 76.33 ± 5.93 bees/5 min, respectively. Strong positive correlations were observed among behavioral and physiological parameters, indicating their interdependence in maintaining colony performance. These findings demonstrate that residual sub-lethal exposure to bifenthrin significantly impairs immunity, longevity, social interaction, and foraging efficiency in A. mellifera, posing serious risks to colony health. Future studies should clarify the mechanisms of these impairments, employ diverse biomarkers, and develop pollinator-safe pest management strategies.
The current study presents a comprehensive morphological and ecological comparison of two orthopteran species, Acheta domesticus (Linnaeus, 1758) and Gryllodes supplicans (Walker, 1859), collected from multiple agroecological zones in Sindh Province, Pakistan. A total of 431 male and 357 female A. domesticus and 282 male and 240 female G. supplicans were examined. Detailed morphological assessments revealed species-specific distinctions, with A. domesticus exhibiting a larger body size and more robust morphological traits compared to G. supplicans. Females of both species were consistently larger than males. Particularly, A. domesticus developed faster, completing its life cycle in approximately 56.7 days, while G. supplicans required around 81 days, suggesting differential life history strategies. Reproductive analysis indicated that G. supplicans follows an r-selected strategy, characterized by high fecundity and a 92% hatching rate, while A. domesticus exhibits traits consistent with K-selection, including higher larval survival and broader environmental tolerance. Thermal and humidity tolerance studies showed that A. domesticus survived across a wider temperature range (15-30°C), whereas G. supplicans preferred narrower, more stable tropical conditions (25-35°C; 60-90% RH). Longitudinal monitoring revealed that G. supplicans exhibited faster population growth, while A. domesticus maintained steady population levels, reinforcing their respective adaptive strategies. These findings emphasize the ecological flexibility of A. domesticus and the reproductive efficiency of G. supplicans, offering valuable insights into species distribution, pest potential, and environmental adaptability. This comparative study enhances our understanding of orthopteran life history evolution and has implications for integrated pest management in agroecosystems under shifting climatic conditions.
Honey bees (Apis mellifera L.), vital agricultural pollinators, face growing threats from insecticide exposure, highlighting the need to understand their trophic-level impacts for sustainable apiculture and crop production. The present study aimed to evaluate the effects of three insecticides viz., flonicamid, pyrifluquinazon, and nitenpyram, on colony strength and honey quality. A sunflower crop was cultivated at four distinct locations, where twelve honey bee colonies were established (three per site). Each colony was either exposed to one of the insecticides or maintained as a pesticide-free control. Standardized protocols were employed to assess adult bee population, brood area, honey yield, and honey quality parameters. The lowest adult bee population (11,874 ± 512.95 bees/colony), brood area (807.14 ± 19.65 cm²), and honey yield (3.44 ± 0.19 Kg/colony) were observed in pyrifluquinazon-treated colonies. In contrast, control colonies exhibited the highest values for adult bee population (20,499 ± 487.31 bees/colony), brood area (1297.8 ± 9.73 cm²), and honey yield (8.24 ± 0.22 kg/colony). Colonies exposed to nitenpyram showed intermediate effects across all measured parameters. A similar trend was observed in honey quality, with pyrifluquinazon-treated colonies producing honey with the lowest enzymatic and antioxidant profiles, followed by flonicamid and nitenpyram. These findings reveal that insecticide exposure disrupts colony health and honey quality, emphasizing the need to avoid pyrifluquinazon and flonicamid during crop bloom and to carefully time nitenpyram applications. Pollinator-safe pest management is essential for sustainable agriculture and food security.
This study evaluated the potential effects of biosynthesized silver nanoparticles (BSN) produced using plant pigments from Clitoria ternatea, Solanum nigrum, Catharanthus roseus (CR), and Cucumis prophetarum at four concentrations: control, 10, 15, and 20 mg L⁻¹. Scanning electron microscopy revealed that the synthesized silver nanoparticles (AgNPs) were predominantly spherical, with an average particle size of approximately 18 nm, as confirmed by particle size distribution analysis. Fourier transform infrared spectroscopy indicated the presence of O–H (carboxylic acid) and N–O (nitro compound) functional groups, suggesting that plant biomolecules played a dual role in the reduction and stabilization of AgNPs. X-ray diffraction analysis showed four distinct diffraction peaks at 38.19° (111), 44.37° (200), 64.56° (220), and 77.47° (311), confirming the crystalline nature and high purity of the AgNPs synthesized from the plant extracts. Artificial inoculation of Fusarium oxysporum on gladiolus corms was conducted to assess the antifungal efficacy of BSN. Corms treated with 20 mg L⁻¹ CR-based BSN exhibited dark brown lesions with approximately 90% disease suppression, reduced fungal colony size (2.36 cm and 2.14 cm), and significantly extended vase life (20 and 21 days) in the ‘White Prosperity’ and ‘Red Advance’ cultivars, respectively. BSN-coated corms stored under ambient and cold storage conditions showed minimal disease incidence, with reductions of 1.56% and 1.05% in ‘White Prosperity’ and ‘Red Advance’, respectively, at 20 mg L⁻¹ CR treatment. Overall, BSN, particularly CR-based nanoparticles at 20 mg L⁻¹, proved highly effective in controlling Fusarium wilt and enhancing vase life, highlighting their potential as an eco-friendly strategy for crop protection and postharvest management in floriculture.
Conocybe telasar sp. nov. was identified as a distinct species of Conocybe from various locations in district Mansehra, including Telasar, Oghi, Sumb, Balakot, and Attar Sheesha. The novelty of this taxon was established based on morpho-anatomical characteristics and molecular phylogeny of the ITS region, in comparison with already described species retrieved from GenBank. Phylogenetic analysis revealed that our species formed a clade closely related to Conocybe fuscimarginata (JX968238) and C. velutipes (JX968233). It is distinguished from allied species by several morphological features, such as creamy white basidiomata with central discoloration, white stipe arising from non-fragile ground grassland habitat, smooth pileus lacking dust and hairs, and ovate-elliptical spores measuring 7.5-10 × 4-6 µm. Microscopic observations showed cheilocystidia measuring 20-33 × 6.2-8.9 µm, while pleurocystidia were absent. Both morpho-anatomical and molecular evidence support the novelty of this species. Detailed descriptions and comparisons with allied taxa are provided in the results section.
Euconocephalus pallidus (Redtenbacher, 1891), a katydid species belonging to the family Tettigoniidae, plays a significant ecological role across diverse habitats in Pakistan. Despite its ecological importance, molecular data on its genetic diversity and evolutionary relationships remain scarce. To address this knowledge gap, the present study investigates the genetic variation of E. pallidus using DNA barcoding of the mitochondrial Cytochrome c Oxidase I (COI) gene. PCR amplicons were purified and sequenced unidirectionally using the Sanger sequencing method. Raw sequences were processed, aligned using ClustalW, and compared with reference sequences from the NCBI database via BLAST analysis. Phylogenetic relationships were inferred using the Maximum Likelihood method based on the Tamura-Nei model in MEGA11 software. BLAST results showed high sequence identity between the obtained COI sequences and those of closely related species, supporting the morphological identification of E. pallidus. The resulting phylogenetic tree placed E. pallidus in a clade with congeners such as E. nasutus and E. varius, supported by moderate bootstrap values. Furthermore, the observed intra-population genetic variation suggests potential phylogeographic structuring within the sampled population. This study provides novel insights into the genetic diversity and evolutionary positioning of E. pallidus in Pakistan. It represents the first molecular assessment of this species using DNA barcoding in the ecological context of the Bahawalpur division, addressing a critical gap in molecular data for Orthopteran insects in the region.
The present study aimed to evaluate the insecticidal efficacy of alkaloid extracts from Duranta repens fruits at concentrations of 4%, 6%, and 8% against different developmental stages of the southern cowpea beetle (Callosobruchus maculatus) under laboratory conditions. The tested stages included eggs, last instar larvae, and pupae at 24 and 120 h of age. The 8% alkaloid extract exhibited the highest ovicidal activity, achieving 100% egg mortality, compared to 60% mortality at the 4% concentration. Similarly, the 8% concentration induced the highest larval mortality (82%) in the last instar stage, significantly outperforming the lower concentrations. Pupae treated at 24 h old with the 8% extract recorded a 93% mortality rate, while 120-h-old pupae showed a reduced mortality rate of 77% at the same concentration. In contrast, the 4% extract resulted in 68% and 60% mortality for 24- and 120-h-old pupae, respectively. Furthermore, the 8% alkaloid extract was most effective in protecting stored cowpea seeds, recording the lowest seed loss (6.55%) after six months of storage, compared to the 4% and 6% treatments. The study also demonstrated that seed germination was not adversely affected by any treatment across all evaluation periods (1 day, 90 days, and 180 days). Overall, the 8% alkaloid extract of D. repens fruits proved to be the most effective concentration in controlling C. maculatus and preserving seed viability during storage.
Sucking insect pests, including Bemisia tabaci (whitefly), Amrasca devastans (jassid), and Thrips tabaci (thrips), cause substantial yield losses in cotton, with reported reductions of up to 40%. To evaluate effective management options, a field experiment was conducted in Muzaffargarh, South Punjab, Pakistan, to assess the toxicity of five insecticides, Nitenpyram, Acephate, Amidecloprid, Buprofezin, and Diafenthiuron, against Bt cotton (Gossypium hirsutum L. cv. IUB-B). The experiment was laid out in a randomized complete block design with three replications, and pest populations were recorded at 24, 48, and 72 h after insecticide application. The results revealed that all insecticidal treatments significantly reduced pest populations compared with the untreated control (p 0.05). Among the tested insecticides, Acephate exhibited the highest overall efficacy, reducing B. tabaci populations to 3.64 individuals at 48 h compared with 15.56 individuals in the control, and completely suppressing T. tabaci populations by 72 h (control: 4.02 individuals). Diafenthiuron was particularly effective against A. devastans, reducing its population to zero within 48 h, compared with 14.9 individuals in the untreated plots. Nitenpyram and the remaining insecticides also caused significant population reductions, though to a comparatively lesser extent. These findings demonstrate the strong field efficacy of Acephate and Diafenthiuron against major cotton sucking pests. However, prolonged reliance on chemical insecticides may accelerate resistance development and pose environmental risks. Therefore, the integration of these insecticides into an Integrated Pest Management framework, incorporating insecticide rotation, biological control agents, and reduced chemical inputs, is strongly recommended to ensure sustainable and long-term pest management in cotton production systems.
