IndexNatural enemiesHymenoptera parasitoidsPredatorsFungal control against bark beetlesPinus Wallichiana branches used in experimentsFungal species used in treatmentFungal treatment of bark beetle adults (Petri dish test)PhotographyStatistical analysisNatural enemiesStudies on important enemies natural species associated with bark beetles were followed according to the standard methods adopted by Dahlsten and Stephen (1974) and Narendran et al. (2001) with minor adjustments. Say no to plagiarism. Get a tailor-made essay on "Why Violent Video Games Shouldn't Be Banned"? Get an original essay Hymenoptera parasitoids To record the seasonal incidence of hymenoptera parasitoids associated with bark beetles, namely I. stebbingi, P. major and P. scitus, experiments were conducted during 2015-2016 in Nowpora village ( 33°61.078' N, 075°18.700' E, altitude 5920 feet) in Anantnag district, Jammu and Kashmir. The samples were collected over a two-year period (April to November 2015 and April to November 2016). After each month, ten branches (1–8 cm in diameter, 20–50 cm long) were cut by careful observation from host trees (P. wallichiana) naturally infested with bark beetles. The sample branches were taken to the laboratory and stored in breeding boxes made of glass equipped with a white muslin sheet for the possible appearance of parasitoids (Figures 2-4). After every ten days, the parasitoids emerging from the infested branches were counted and some branches were also debarked to examine the activities of the parasitoid stages associated with the bark beetles. The same procedure was followed throughout the year and the collected parasitoids were listed and the proportion of a particular species out of the total (i.e. the dominance coefficient) was determined. The dominance coefficient of a parasitoid species was calculated as follows:During the year 2016, the same sampling procedure was followed and the dominance coefficient (%) was determined according to the above formula.PredatorsTo record the seasonal presence of predators associated with bark beetles, namely ., I. stebbingi, P. major and P. scitus, experiments were conducted in the same study area mentioned above. Samples were collected over a two-year period (April to November 2015 and April to November 2016) with an interval of 15 days between subsequent samplings. Since the three bark beetle species mentioned above occupy different parts of the host tree, three wooden structures (sampling units) were made available, one for each species. The dimensions of the wooden frames were 0.06 m2, 0.10 m2 and 0.16 m2 for P. scitus, P. major and I. stebbingi, respectively. Every fortnight, a total of thirty samples (ten for each species) were taken from severely infested trunks, first marking the surface of the bark using wooden frames, then carefully debarking the sample area occupied by each species (Fig. 3– 4). Data for each predator species with its associated host beetle stage were recorded. The same procedure was followed throughout the year and the collected predators were listed and the proportion of a particular species in the total (i.e. dominance coefficient) was determined according to the above-mentioned formula used for parasitoids in both years. Fungal control studies against bark beetles on the use of entomopathogenic fungi against bark beetles was followed according to previous standard methods adopted by Batta (2007) and Jakus and Blanzee (2011) Pinus Wallichiana branches used in experiments Branches naturallyinfested with P. wallichiana were collected during 2017 (April to November 2017) from a severely infested pine forest located in Nowpora village (33°61.078' N, 075°18.700' E, altitude 5920 ft) in Anantnag district , Jammu and Kashmir (Figure 1) and forest check point, Tangmarg (340 03.797' N, 074024.948' E, elevation 7552 ft) in Baramulla district, Jammu and Kashmir (Figures 1¬¬¬¬–3). The infested branches were then selected by observing bark beetle infestations (Figure 4–5). The sample branches were transported to the Animal House, Department of Zoology, Kashmir University in plastic boxes for evaluation of fungal treatments against I. stebbingi. Fungal species used in treatment Commercial bioprepration of three entomopathogenic fungi, namely B. bassiana, M. anisopliae and L. lecanii were obtained from Green Life Biotech Laboratory, Somanur, Coimbatore, India. The experiment was performed from April to November 2017. A total of 90 branches naturally infested with bark beetles, classified into five groups (G1–G5), for each bark beetle species, were used in the experiment. Each replicate represented three infested branches, and six replicates per experimental treatment were used for each bark beetle species (Table 1). The insecticide used was cyclone (active ingredient: Chlorpyrifos 50% + Cypermethrin 5%). The fungal preparation was diluted in water: 1 ml of biopreparation/1000 ml of water with four drops of a common detergent as a wetting agent. Each fungal suspension contained 1.0 x 109 fungal spores in 1 ml. Fungal suspensions were applied with a hand sprayer at 500 ml per log (Table 1). For effective treatment, high volumes of fungal suspensions were used so that the suspensions penetrated spontaneously after application. After 10 days, nine branches of three treated replicates in each group were carefully debarked, and the percentage mortality of each bark beetle species was calculated and compared (Table 1). The same procedure was applied to calculate the percent mortality of each bark beetle species after 20 days of treatment. Fungal Treatment of Bark Beetle Adults (Petri Dish Test) A total of 15 Petri dishes containing filter papers were used in this method; three replicates were maintained for each treatment. Treatments were performed by applying two rapid jet sprays standardized to 1.0 ml per replicate using a small calibrated hand sprayer (1 liter capacity) equipped with a nozzle suitable for low volume spray application (Batta, 2007). 40 adults of each bark beetle species were introduced into each Petri dish before spraying. The same spray volumes (1 ml per replicate) were applied in the other treatments (Table 2). The mortality rate of each treated group was evaluated 2, 4 and 6 days after treatment. This mortality was demonstrated by the lack of movement of treated adults within a five-minute period of continuous observation or by the appearance of micellar growth on the bodies of dead adults. The beetles were then incubated in Petri dishes under humid conditions for a week to promote micellar growth with conidia and conidiophores on their bodies. PhotographyPhotographs during the field study were taken using the Canon PowerShot SX60 camera equipped with a macro lens (Raynox MSN-505, 37mm). Digital image analysis was performed using ImageJ analysis software (version 2006.02.01). For the morphometric description of the collected predators, images were taken from the haplotype with a Leica DFC295 camera connected to 10.00).