Rising concerns over chemical insecticides are demanding alternative pest management methods. The present study planned to explore the potential of elemental-ES, bio-sulphur-BS, compost-Cp, and compared to insecticide (Carbosulfan) and untreated controls for sustainable management of canola aphids (Brevicoryne brassicae). Split doses were more useful (screen house experiments) over single dose (in field). BS mixed with Cp provided best results to suppress aphid reproduction. Insecticide effects were instant but not long-lasting. Sulphur mixture also improved crop yield and oil contents; however, plant height was maximum in insecticide. RP-HPLC revealed activation of different phenolic defense chemicals (three flavonoids, five phenolic acids, and six hydroxycinnamic acid) in canola. Phenolic compounds were higher in sulphur-treated plants but their distribution patterns were altered in leaves, shoots, and seeds. The results could be extended to insect pests on other crops. However, further genetic studies are needed to discover the intricate pattern of sulphur resistance.

Climate change is among the greatest drivers of biodiversity loss, threatening up to 15–30% of described species by the end of the twenty-first century. We estimated the current suitable habitat and forecasted future distribution ranges of Indian pangolin (Manis crassicaudata) under climate change scenarios. We collected occurrence records of Indian pangolin using burrow counts, remote camera records and previously published literature in Pakistan during 2021–2023. We downloaded bioclimatic data for current (1970–2000) and future (2041–2060, 2061–2080, 2081–2100) climate scenarios from the WorldClim database using the Hadley Global Environment Model (HadGEM3-GC31-LL). We used MaxEnt software to predict current and future distributions of Indian pangolin, then computed the amount of habitat lost, gained, and unchanged across periods. We obtained 560 Indian pangolin occurrences overall, 175 during the study, and 385 from our literature search. Model accuracy was very good (AUC = 0.885, TSS = 0.695), and jackknife tests of variable importance showed that the contribution of annual mean temperature (bio1) was greatest (33.4%), followed by the mean temperature of the coldest quarter (bio-12, 29.3%), temperature seasonality (bio 4, 25.9%), and precipitation seasonality (bio 15, 11.5%). The maxent model predicted that during the current time period (1970–2000) highly suitable habitat for Indian pangolin was (7270 km2, 2.2%), followed by moderately suitable (12,418 km2, 3.7%), less suitable (49,846 km2, 14.8%), and unsuitable habitat (268,355 km2, 79.4%). Highly suitable habitat decreased in the western part of the study area under most SSPs and in the central parts it declined under all SSPs and in future time periods. The predicted loss in the suitable habitat of the Indian pangolin was greatest (26.97%) under SSP 585 followed by SSP 126 (23.67%) during the time 2061–2080. The gain in suitable habitat of Indian pangolin was less than that of losses on average which ranged between 1.91 and 13.11% under all SSPs during all time periods. While the stable habitat of the Indian pangolin ranged between 64.60 and 83.85% under all SSPs during all time periods. Our study provides the current and future habitat ranges of Indian pangolin in the face of a changing climate. The findings of our study could be helpful for policymakers to set up conservation strategies for Indian pangolin in Pakistan.

Sesame (Sesamum indicum L.), a member of the Pedaliaceae family, is referred to as the queen of oilseeds. There are numerous barriers to its production and productivity, including a lack of economically feasible and technically adequate manufacturing technology, but insect pests particularly, sesame leaf webber (Antigastra catalaunalis) and Mirid Bug (Creontiadus dilutes) play a prime role in declining its potential yield. In this study, five treatments, viz. T1:2-WAE (weeks after emergence), T2:2,4-WAE, T3: 2,4,6 WAE, T4: 2,4,6,8 WEA, T5: weekly spray after emergence (positive control) and T6: control (zero application of spray) spray were applied, while T5=weekly spray started one week after emergence, prior to the onset of A. catalaunalis infestation on the sesame crop and was continuously applied until the maturity of the crop. A Hand knap sack sprayer was used for the Polytrin C 440EC (Profenophos 400 G/L + Cypermethrin G/L) @500ml/acre application. The results revealed that the incidence of A. catalaunalis and C. dilutes throughout the season during both years (2022-23) was significantly different (p < 0.001) among all treatments. High insecticide application frequency lowers the leaf, flower and capsule damage, which ultimately decreases seed loss and enhances the end product, so it was observed that T5: weekly spray after emergence (positive control) gave maximum control, followed by four-time spray application (T4: 2, 4, 6, 8WEA) while maximum infestation was observed in T6: control (zero application of spray), followed by T1:2-WAE. Positive control gave the best results. It is suggested that three-time spray application was found to be most suitable for the management of both devastating insects. Based on evaluation, it appears that adjusting the frequency of insecticide application can effectively control sesame leaf Webber and mirid bug populations. However, it’s important to balancer pest control with environmental concerns and resistance development.

Fruit flies are among the most destructive pests, causing significant economic losses worldwide, including in Pakistan. Various management strategies have been explored, with entomopathogenic nematodes (EPNs) emerging as promising biocontrol agents. This study evaluated the effectiveness of two EPNs, Steinernema glaseri and Heterorhabditis bacteriophora, along with two insecticides, lufenuron and bifenthrin, against Bactrocera dorsalis under laboratory conditions. EPNs were cultured and reared in the lab on the last larval instar of Galleria mellonella. Fruit fly larvae were collected from the field and reared in the lab to ensure a sufficient population for bioassays. Different concentrations of insecticides and EPNs alone and in combination were applied to larvae and pupae of B. dorsalis. Both insecticides and EPN revealed higher mortality rates against fruit flies with increased exposure time and higher concentrations. Alone treatment of both insecticides produced 100% of the mortality of both larvae and pupae at their highest concentrations. The maximum mean mortality of individual larvae (9.75 individuals) and pupae (10.00 individuals) have been produced by H. bacteriophora. While S. glaseri produced the mean mortality of 9.00 individual larvae and 9.25 individual pupae after 144h of treatment application. The combined treatments of insecticides and EPNs also caused mortality, the effectiveness was comparatively lower than that observed with individual treatments of either insecticides or EPNs. The maximum mean mortality has been produced by H. bacteriophora+ bifenthrin 8.75 individual larvae and 9.25 individual pupae. These results suggest that synergistic or additive effects may occur under certain conditions.

The SPL gene family (for Squamosa Promoter-binding like Proteins) represents specific transcription factors that have significant roles in abiotic stress tolerance, development and the growth processes of different plants, including initiation of the leaf, branching and development of shoot and fruits. The SPL gene family has been studied in different plant species; however, its role is not yet fully explored in pigeon pea (Cajanus cajan ). In the present study, 11 members of the CcSPL gene family were identified in C. cajan . The identified SPLs were classified into nine groups based on a phylogenetic analysis involving SPL protein sequences from C. cajan , Arabidopsis thaliana , Cicer arietinum , Glycine max , Phaseolus vulgaris , Vigna unguiculata and Arachis hypogaea . Further, the identification of gene structure, motif analysis, domain analysis and presence of cis -regulatory elements in the SPL family members were studied. Based on RNA-sequencing data, gene expression analysis was performed, revealing that CcSPL2.1, 3 and 13A were significantly upregulated for salt-tolerance and CcSPL14 and 15 were upregulated in a salt-susceptible cultivar. Real-time qPCR validation indicated that CcSPL3, 4, 6 and 13A were upregulated under salt stress conditions. Therefore, molecular docking was performed against the proteins of two highly expressed genes (CcSPL3 and CcSPL14 ) with three ligands: abscisic acid, gibberellic acid and indole-3-acetic acid. Afterward, their binding affinity was obtained and three-dimensional structures were predicted. In the future, our study may open avenues for harnessing CcSPL genes in pigeon pea for enhanced abiotic stress resistance and developmental traits.

Red spider mite, Tetranychus urticae Koch, causes severe damage to economically important crops, fruits, vegetables, ornamentals etc. Management of red spider mite largely depends on chemicals. Application of insecticides may cause massive outbreaks of mite population. Sublethal effects of imidacloprid, thiamethoxam and acetamiprid doses were investigated on life-table parameters of red spider mite. All neonicotinoids revealed significant effects on life characteristics i.e. development duration and fecundity of red spider mite. Mites treated with imidacloprid, thiamethoxam and acetamiprid significantly reduced the net reproductive rate (Ro). No significant difference in finite rates of increase (λ) was observed among the treatments. Imidacloprid reduces the egg duration and mean generation time (T). Emerged adult longevity was shorter when exposed to insecticides. The total adult longevity was significantly reduced by the insecticides in comparison with the control. Highest mean generation time was observed in acetamiprid treated mites. The results suggested that sublethal effects of tested insecticides affected the biological parameters of red spider mite.

The CRISPR/Cas system has been recognized as a transformative tool in cereal improvement, offering precise and efficient gene editing capabilities. Pathogens and pests trigger biotic stress, posing a significant danger to agricultural productivity and food security. Gene knockouts can disable vulnerabe genes, while gene editing enables the incorporation of naturally occurring resistance genes from other plant species. Additionally, gene regulation can fine-tune the expression of stress-responsive genes, bolstering the plant's ability to fend off biotic stressors. Beyond the well-known CRISPR/Cas9 system, there exists another CRISPR effector (class 2) called Cas12a or Cpf1, belonging to the V type CRISPR/Cas system. Unlike Cas9, Cas12a lacks the HNH domain but contains the RuvC domain. Further, there are three separate class 2 CRISPR systems; C2c1, C2c2, and C2c3. Among them, presence of RuvC-like endonucleases in C2c1 and C2c3 categorized them as type V-C (Cas12c) and V-B (Cas12b), respectively. Whereas, C2c2 (Cas13) stands out with its unique properties and designated as class 2 type VI. These discoveries in diverse CRISPR systems hold significant implications for cereal biotechnology and crop improvement. Despite these promising prospects, it is essential to consider regulatory and societal aspects to ensure the responsible and sustainable deployment of this biotechnological approach in agriculture. By combining CRISPR with other genetic technologies, it may be possible to engineer cereals that are more climate-resilient and sustainable, addressing the ever-growing global demand for food and contributing to food security and agricultural sustainability.

Leucinodes orbonalis is a potential pest of eggplant, commonly known as fruit and shoot borer. Owing to its devastating nature, chemical pesticides are frequently applied and insecticide pressure is significantly higher in developing nations. Keeping the current situation in mind the most effective and practical strategy for managing insect pests found in humid and subtropical environments is the bio-control. Entomopathogenic nematodes, has been utilized against many pests for their effective management. Keeping in view the biocidal potential of entomopathogenic nematodes, the current study was planned to explore the biocidal potential of Steinernema and Heterorhabditis, two significant genera of entomopathogenic nematodes against L. orbonalis. White trap extraction technique was used to isolate nematodes from several insect larvae. Four discrete concentrations of entomopathogenic nematodes viz. 60, 90, 120, and 150 infective juveniles per larvae were applied on larval stages. The virulence trial was followed by completely randomized block design. Multiple comparisons showed that among both the tested nematodes H. bacteriophora induced significant 81.33% mortality corresponding to 150 IJs/larva after 72 h of exposure as compared to S. glaseri. In the view of current research, entomopathogenic nematodes are recommended to be used in integrated pest management program for the management of insect pests.

A laboratory study was carried out in which wheat flours of five conventional non -fortified cultivars (AAS-2011, Gold, Manthar, Fareed-2006 and Miraj-2008), one zinc fortified cultivar (Zincol) and two non -fortified lines (1620 and 1091) were offered to adults of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) to assess their oviposition, larval, pupal and adult emergence and developmental durations of eggs, larvae, pupae and adults. The results revealed that the lowest ovi position was recorded on Zincol, while the highest fecundity was recorded on flour of cultivar Gold. Contrarily, the incubation period of T. castaneum eggs was extended on Zincol flour as compared to the rest of the cultivars and lines. Similar trends were also observed for the number of hatched larvae, larval period duration, pupation, pupal period duration, adult emergence, and adult long evity. Overall, Zincol flour red uced the production of eggs, larvae, pupae and adults of T . castaneum and extended their developmental durations. Our study suggested that zinc-fortified wheat cultivar negatively affected the reproductive potential of T. castaneum and h inders its development. Hen ce, it is concluded that zinc-fortified wheat cultivars can play important roles in integrated pest management (IPM) of wheat for long -term storage.

Niche overlap between sympatric species can indicate the extent of interspecific competition. Sympatric competing species can exhibit spatial, temporal, and dietary adjustments to reduce competition. We investigated spatial, temporal, and dietary niche overlap of sympatric Asian palm civet (Paradoxurus hermaphroditus) and small Indian civet (Viverricula indica), in and around Pir Lasura National Park, Pakistan. We used remote cameras to determine the frequency and timing of detections to estimate spatial and temporal overlap, and prey remains from scats to estimate dietary overlap. We collected scat samples of Asian palm civet (n = 108) and small Indian civet (n = 44) for dietary analysis. We found low spatial (Oij = 0.32) and temporal (Δ = 0.39) overlap, but high dietary niche overlap (0.9) between these two civet species. Both civet species were detected at only 11 camera sites and small Indian civets were detected most frequently during 2:00–5:00 h and 8:00–10:00 h, whereas Asian palm civets detections were greatest during 20:00–2:00 h. The overall niche breadth of Asian palm civet was slightly narrower (L = 9.69, Lst = 0.31) than that of the small Indian civet (L = 10, Lst = 0.52). We identified 27 dietary items (15 plant, 12 animal) from scats of Asian palm civet including Himalayan pear (Pyrus pashia; 27%), Indian gerbil (Tatera indica; 10%), Rhesus monkey (Macaca mulatta; 4%), and insects (5%). Scat analysis of small Indian civets revealed 17 prey items (eight plant, nine animal) including Himalayan pear (24%), domestic poultry (15%), Indian gerbil (11%), and house mouse (Mus musculus; 5%). Both civet species consumed fruits of cultivated orchard species. Spatial and temporal partitioning of landscapes containing diverse foods appears to facilitate coexistence between Asian palm civets and small Indian civets.

Nanotechnology is a well advanced area and different types of nanoparticles are now produced. The most revolutionary topic of the twenty-first century is nanotechnology. Recently, metallic nanoparticles (NPs) have gained a lot of attention. In this study a straightforward, safe, and economical method is used for synthesizing copper nanoparticles (CuNPs) by using cell free filtrate of Aspergillus niger. Fungi secretes enormous amount of enzymes which act as reducing as well stabilizing agents. Synthesized CuNPs were characterized by different techniques. Ultraviolet (UV)-visible spectrophotometer showed a characteristic peak at 380nm, Dynamic Light Scattering (DLS) analysis indicated that CuNPs have average size of 570nm with 0.480 PDI. SEM (Scanning Electron Microscopy) revealed CuNPs are of cubic shape. As per FTIR (Fourier Transform Infrared Analysis) two functional groups i.e., -O-H- and C=C- on the surface of CuNPs. Genetic analysis of Aspergillus niger revealed the presence of nitrate reductase enzyme that may involve in reduction of metal into NPs. Furthermore, CuNPs showed significant antibacterial action against Escherichia coli, Bacillus subtilis, Staphylococcus aureus, and Klebsiella pneumoniae with 17mm, 20mm, 17mm and 20mm respectively. CuNPs also showed antioxidant potential with 59.41µg/ml. Fungi secretes valuable enzymes with inherent ability to reduce metal ions into NPs. This reduced the cost of NPs synthesis and also synthesized NPs exhibit greater biomedical applications.

Cotton is an important cash crop of Pakistan and export commodity.Yield of cotton crop is affected by many factors but insect pests especially whitefly Bemisia tabaci (Homoptera: Aleyrodidae) is destroying cotton crop yield and quality in recent times.Many control measures are adopted against this pest including chemical control methods.Due to certain environmental issues and insecticide resistance problems, adoption of alternate methods of insect control is imperative.In this experiment we used some insecticides alone and in combination with Salicylic acid against B. tabaci.Flonicamid 50% DF at the rate of 80 g/ac and Diafenthiuran 50% SC at the rate of 200 mL/ac were used against Whitefly.Salicylic acid (SA) by Sigma-Aldrich 500 G was used 01 mL/L of water along with the tested insecticides under field conditions.Highest reduction 90.63% was observed in Flonicamid+SA treatment followed by 82.89% in Diafenthiuran +SA, Flonicamid 77.76%, Diafenthiuron 76.28% and lowest 29.36% reduction was observed in SA treatment with respect to control after 72 hrs. of treatment application.After 7 days maximum reduction 83.43% was observed in Flonicamid+SA followed by 69.02 % in Diafenthiuron+SA, 77.76% in Flonicamid, 66.37% in Diafenthiuron and lowest 21.97% recorded in SA treatment.In SA treatments population of rove beetle, green lace wing and spiders was significantly higher as compared to other treatments.It is evident from the results that SA has enhanced plant resistance against whitefly and thus reduced the damage of pest.SA can be used as alternative combination for the management of cotton whitefly and the amount of insecticide can be reduced for the control of whitefly.

The study was aimed to isolate and identify the methicillin-resistant strains and then detect the genetic variants and investigate S. aureus isolates that were resistant to methicillin found in the community of Southern Punjab. Collecting the isolates of S. aureus from the Southern Punjab region of Pakistan. Isolation and identification of these collected isolates were done by subjecting these isolates to laboratory procedures. A polymerase chain reaction was performed for the molecular and genetic analysis. 60 urine and 40 blood samples were taken from outdoor and indoor patients of the Nishtar Medical College & Hospital, Multan. Gram staining and different biochemical assays were done to confirm the presence of S. aureus. After the confirmation of S. aureus, DNA extraction was performed by a modified method of CTAB. A polymerase chain reaction was performed to analyze the size of amplicons found in the Southern Punjab community. In order to check the resistance and susceptibility pattern of S. aureus against beta-lactam antibiotics and fluoroquinolones, the Kirby-Bauer method was used. Out of 100 samples, 98 were cultured on blood agar and mannitol salt agar. 92 tested gram-positive and out of which only 88 gave positive results for the catalase test. When a coagulase test was performed, 85 produced coagulations with plasma in the test tubes. Upon antibiotic susceptibility testing, 50 samples were found as methicillin-resistant S. aureus.67% of methicillin-resistant S. aureus contains mecA3, femA3, aac(6’)/aph(2’’), Tet(K)13, Tet(M)13 genes and methicillin-susceptible S. aureus has 33% less prevalence as compared to methicillin-resistant S. aureus. Southern Punjab region of Pakistan was found to possess the genes mecA3, femA3, aac(6')/aph(2''), Tet(K)13, and Tet(M)13. Southern Punjab region outnumbered in Methicillin-resistant S. aureus(MRSA) isolates in terms of the prevalence of mecA3, femA3, aac(6’)/aph(2’’), Tet(K)13, Tet(M)13 genes. Non-beta lactam antibiotics can be used to treat MRSA infections.

Tribolium castaneum is devastating cosmopolitan insect pest of stored grain. The present study investigated the impact of three insect growth regulators on different life stages of T. castaneum. Percent mortality (%) induced was observed at 0.75. 1.25, 2.5, 5, 10, and 20 ppm doses of each IGR after 7, 14 and 21 day exposure. In addition to larval and parental mortality, progeny suppression data was also recorded. The larvae of the 2nd instar were more susceptible to lufenuron, with mortality reaching 100% at 10ppm even after a 14-day interval. In the case of pyriproxifen, 2nd instar did not responded to all dose rates until 14 days however 21 days, statistically corresponding 2.5 to 10 ppm dose rates was detected. At 20 ppm, methoxy fenazide resulted in 92 percent mortality of 2nd larval instar mortality, and the larvae responded to all dose rates at all exposure intervals that varied significantly among themselves. The mean mortality of the 4th instar larvae at 20 ppm for lufenuron after a 21-day interval was 90%, which was not significantly different from the mortalities (62 and 80%) at the same dose rate after 7 and 14 days, respectively. All the IGRs were very effective in progeny suppression (>80%) and complete progeny suppression (100%) was observed in adults treated with methoxyfenazide and lufenuron at 2.5 and 10 ppm doses, respectively. It is concluded that all the screened IGRs were effective in progeny suppression and in some cases, complete progeny suppression was achieved

Odontotermes obesus (Blattodea: Termitidae) is a prevalent subterranean wood-eating termite species that causes damage to mature trees, saplings and seedlings. The efficacy of most synthetic insecticides against this notorious pest has been compromised primarily because of its enigmatic feeding behavior and development of resistance to a number of insecticides. It has therefore become necessary to explore other alternative biologically sound and low-impact termite control methods, particularly for use in forests. Hence, this study was designed to verify the efficacy of different indigenous EPN isolates (Steinernema carpocapsae, Heterorhabditis bacteriophora and Heterorhabditis indica) against workers of Odontotermes obesus. The pathogenicity of each nematode isolate was assessed in laboratory conditions using filter paper and sawdust bioassay at two different temperatures (16 ± 1 and 26 ± 1 °C). Additionally, the efficacy of the nematode species was also assessed in field conditions. The results of the experiments revealed that the mortality of termite workers was more pronounced in sawdust bioassay in comparison with filter paper bioassay at both the tested temperatures. The mortality response in both bioassays was more pronounced at the higher temperature. A significantly higher mortality was recorded at both tested temperatures for S. carpocapsae followed by H. bacteriophora and H. indica. A dose-dependent positive mortality response was also recorded at both tested temperatures. Similar to the laboratory trials, the field applications of the three tested nematode species showed that maximum mortality was recorded for S. carpocapsae followed by H. bacteriophora and H. indica. It was therefore concluded that indigenous EPNs can provide more effective control of termites, possibly because of their direct interaction with pest species in the soil and the possibility of causing secondary infection through infected cadavers.

High Quality fodder crops are a key component of the white uprising of milk yield. Berseem is a top forage crop also known as the Queen of all Cultivated Fodders or the Green Gold of the Asia. The experiment was designed to screen out the six varieties of berseem viz. F-01-16, F-02-16, F01-17, F-02-17, F-01-18 and F-02-18 against aphid and leaf minor. The data regarding aphid and leaf miner was recorded on weekly bases from five selected plants. The results showed significant difference in aphid and leaf miner population among six berseem varieties. The results depicted that maximum seasonal mean population of aphid was observed in F-02-16 (1.06) followed by F-01-17 (1.03) while minimum mean population was observed in the case of F-02-17 (0.84) followed by F-01-16 (0.96). In case of leaf miner, it was observed that maximum seasonal population was recorded in F-01-18 (13.21) followed by F-02-16 (12.47) while minimum mean population was calculated in the case of F-02-17 (8.45) followed by F-01-16 (11.65).