Publication History
Submitted: October 08, 2023
Accepted: October 11, 2023
Published: December 11, 2023
Identification
D-0170
Citation
Lubna Rasheed, Itrat Hussainy Khan, Nimra, Rooha Farooq, Aqsa Mubeen & Kinza Imtiaz (2023). Evaluation of Beauveria Bassiana and Metarhizium Anisopliae in Integration with Insecticide against Dusky Cotton Bug. Dinkum Journal of Natural & Scientific Innovations, 2(12):765-793.
Copyright
© 2023 DJNSI. All rights reserved
765-793
Evaluation of Beauveria Bassiana and Metarhizium Anisopliae in Integration with Insecticide against Dusky Cotton BugOriginal Article
Lubna Rasheed 1*, Itrat Hussainy Khan 2, Nimra 3, Rooha Farooq 4, Aqsa Mubeen 5, Kinza Imtiaz 6
- University of agriculture, Faisalabad, Pakistan; lubnarasheed0@gmail.com
- University of agriculture, Faisalabad, Pakistan; itrathussaink@gmail.com
- Lahore College for Women University, Lahore, Pakistan
- Lahore College Women University, Sialkot, Pakistan; roohafarooq0007@gmail.com
- Lahore College for Women University, Lahore, Pakistan; aqsamubeen2130@gmail.com
- Lahore College for Women University, Lahore, Pakistan; kinzaimtiaz156@gmail.com
* Correspondence: lubnarasheed0@gmail.com
Abstract: Oxycarenus hyalinipennis has becomes serious cotton crop pest in Pakistan during the recent years. In Pakistan cotton is cash crop and the most significant exportable crop. Its fiber is an important raw material for many textile industries as it accounts for more than 60% of all textile fibers. In the present study, evaluation of Metarhizium anisopliae and Beauveria bassiana in integration with the pesticide against dusky cotton bugs was assessed. Adults of the dusky cotton bugs were collected from different field of cotton of the University of Agriculture Faisalabad. Collected specimens were reared in the Biological Control Lab, Department of Entomology University of Agriculture Faisalabad under controlled lab conditions. The efficacy of these microbial agents could be enhanced by applying in combination with insecticides. Two concentrations i.e., 1×106, 1×108 conidia/ml of each entomopathogenic fungi were used in combination with insecticides against 5th instar nymphs of O. hyalinipennis. Complete randomized design (CRD) was used with ten treatment and three replications and each treatment applied homogenously in all the boxes simultaneously. Data collection was started after the application of treatments. The maximum mortality in 5th instar nymph of Oxycarenus hyalinipennis (98.33%) was observed on day 4 in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (83.33%). And the maximum adult mortality of Oxycarenus hyalinipennis was observed on day 6 in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) (100%) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (96.67%). The combined treatment of entomopathogenic fungi and synthetic insecticides caused higher mortality of O. hyalinipennis. There was no significant population reduction recorded in control conditions. The results indicated that these entomopathogenic fungi can provide better control in combination with insecticides and can be used in integrated pest management for reducing the population density of O. hyalinipennis.
Keywords: Beauveria Bassiana, Metarhizium Anisopliae, Insecticide, Dusky Cotton Bug
- INTRODUCTION
Dusky cotton bug (Oxycarenus hyalinipennis) which belongs to the family malvaceae is a major economic insect pest of cotton (El-Rahim et al., 2015). The nymphs and adults of dusky cotton bug get moisture and food from the leaves and seeds of cotton plants (Khan and Ahmed, 2000). The O. hyalinipennis is an emergent threat to the cotton industry of Pakistan. Although in various countries this is a most important insect pest of cotton crop which is responsible for the loss in cotton crop. Dusky cotton bug starts nourishing, reproducing and laying eggs on the mature seeds of cotton. Adults and nymphs absorb oil from the seeds. On the cotton lint female lay eggs and get their feed from the seed (Sweet, 2000). Dusky cotton bugs are dark brown in color with transparent wings. Length of the each adult is about 4 to 5 mm. Nymphs are smaller and have rounded abdomen when they mature look like adults. Female of dusky cotton bug laid eggs either separately or in 2 to 10 groups in lint of partially opened cotton bolls and on seeds. Newly laid eggs of dusky cotton bugs are white then change into the pale color, finally to the light pink in color before the hatching. Color of newly hatched adults is bright red. Females have longer life cycle than males. Life cycle of male is about 33 to 49 days with a usual of 38.95 days as compared female life cycle is about 35 to 51 days with a usual of 41.10 days in lab conditions. Seed are not ripen, lose color and remain light in weight when sap is sucking by nymph and adults of dusky cotton bug (Kedar et al., 2014). In recent years dusky cotton bugs are the major insect pest of the cotton crop in Pakistan. It causes both quantitative and qualitative damages to cotton in several cotton growing countries Worldwide (Atta et al., 2015). Cotton strainer has been identified as one of the crucial pest of cotton in several regions of the world. The damage caused by the bugs on unripe seeds was therefore negligible. Now, however, with the introduction of Bt cotton, the crop span lasts for the whole year. On the other hand, the use of contact poisons has become limited and there is total reliance on systemic insecticides. Under such conditions, the dusky cotton bug was able to flare up, attaining the status of a major pest. It causes a continuous decrease in cotton-seed weight, germination and oil quality (Henry, 1983). Major injury is done in seed by decreasing value, oil content and germination (Sweet, 2000). Adults and nymphs suck sap and moisture from the leaves and seeds (Khan and Ahmed, 2000). Nymphal stages of dusky cotton bug and the adults nourish on a seed in emerging bolls of cotton, making the lint muddy in appearance. Dusky cotton bug feed by piercing the young cotton bolls or the flower buds typically cause decrease in the size and fruiting body might abandon or drop to ground. The damaging behavior of dusky cotton bug disrupts the cotton field at every stage. Cell sap is extracted from the reproductive parts of plants and it also destroys the lint quality. Also injurious the kernels and reproductive parts it also worsens the lint value causing in poor ginning of the cotton fibers. Its massive population is actuality noted in squares subsequent in alteration of the square’s color from olive green to pale creamy and eventually flaking of the criticized squares. Former it was thought that DCB lie dormant un-ginned cotton in ginning factories and houses. (Shah et al., 2016). In Pakistan cotton is cash crop and the most significant exportable crop. Its fiber is an important raw material for many textile industries as it accounts for more than 60% of all textile fibers. It is most important source of foreign exchange for the country. Cotton is major source of income for underprivileged inexperienced people and for farmers especially women. Population of the rural areas consist of more than two third of the country population. In agricultural sector cotton contributes 7% to the national economy in the value added (Farooq, 2014). For last few years dusky cotton bug was stated as a severe insect pest of the cotton crop which caused weight reduction in cotton seed but reduced oil quality and germination. Large population of dusky cotton bug feeds on seed of cotton can cause 88% losses in seed germination. This pest decreased 6.8% yield of cotton, 32% oil content and 6% seed weight When population of DCB was fifty pair per boll it cause damage of 42.9% in cotton seed weight, 35.1% of oil content and 29.3% in seed germination (Srinivas and patil, 2004). The color of the cotton lint turned yellow during ginning process. Due to the attack of dusky cotton bug, cotton seed is normal in shape from the outside but from inside embryos are wasted, discolored and 15% loss in weight can occur (Schaefer and panizzi, 2000). Dusky cotton bug has also been reported feeding on certain vegetables and fruits including corn, dates, avocado, grapes, okra, peach, pomegranate and figs (USDA, 2010). Current method used to control dusky cotton bug is to use pesticides that cause serious problem to natural environment. There is a need to develop alternative strategies such as biological control use of entomopathogenic fungi with new chemistry pesticide as potential management strategy. Entomopathogenic fungi have an endophytic capacity that induces a wide range of changes in the defensive compounds and composition of plant nutrients (Mas et al., 2019). Special set of insects pathogen are entomopathogenic fungi which are integrated by the host is, no obligation till the contamination outcomes from the perforation of the cuticle. There is a significant event to destroy the the insect pest which involve dealing of the specific disease of the specific insect pest caused by a pathogenic fungi. There is growing evidence that, in addition to causing mortality in insects, they improve plant nutrient uptake, stimulate hormone production and increase tolerance to abiotic and biotic stresses, thereby boosting plant growth (Sasan and Bidochka, 2012). For control of forest insect pest and agricultural pest entomopathogenic fungi have been widely used (Maniania, 1993). Entomopathogenic fungi are available all over the world as a biological control agent for control of an agricultural pest (Wraight et al., 2001) including fungal pathogen Vuillemin and Beauveria bassiana. Against the cotton strainer and the red bug of cotton entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana are used. Beauveria bassiana is well recognized microbial entomopathogen of a diverse range of arthropod species. This species is successfully used as a microbial biocontrol agent for integrated pest management of arthropods pest with many B. bassiana based viable product being available on the market. B. bassiana has the potential to protect plants against various pests (Jackson et al 2010). Metarhizium anisopliae is a well-characterized entomopathogen (Santi et al., 2010). Entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (Deuteromycotina: Hyphomycetalis) is a rather common agent causing infection in natural insect populations (Ogarkov and Ogarkova, 2000; Borisov et al., 2001; Hughes et al., 2004). Some species of mites and spiders are also observed as the hosts oh this fungus (Brooks et al., 2004; Kirkland et al., 2004).Such a wide dissemination of Metarhizium anisopliae under natural conditions is likely due to both parasitic and saprophytic development of this fungus. As a rule, the transfer from parasitic to saprophytic development is accompanied with sharp change of fungal virulence (Ogarkov and Ogarkova, 2000; Geshtovt, 2002). It has been widely used for the control of various varieties of arthropods including disease vectors and agricultural insect pest (Lazzarini et al 2006). Conidial yield of M. anisopliae is dependent upon the fungal strain and nutrition and does not appear to be linked with radial growth as an increase in radial growth did not resulted in simultaneous increase in conidial yield. For M. anisopliae, and other fungal biocontrol agents (BCAs), to be commercially viable, mass production has to be done on artificial substrates. The artificial use of many organisms effects depressing insect pest’s populations to the bearable bulks and keeps the targets to a less destructive extent (Gronvold et al., 1996). In Pakistan many new chemistry pesticides including pyrethroid and organophosphate are extensively used for the control of cotton pests including dusky cotton bugs (Afzal et al., 2015). Massively operational novel chemistry pesticides against specific insect pests essentially recognized then handled it to reduce danger of a resistance in the insect pest against pesticides. Former research on a susceptibility of the cotton strainer to a numerous pesticides accompanied. Generally six to ten sprays of various pesticides are used on the crop of cotton in a season and two to four sprays on the vegetables e.g. okra (Ishtiaq et al., 2012). The main ketoenol that was commercially accessible, spirodiclofen, Envidor and spiromesifen-Oberon, validated to be fine suited for the use in the modern IPM. Semi-field-trials with useful creatures were supported out to estimate the effect of product under required conditions. Field trainings succeeding Good Agricultural-Practices allowed the-assessment of the Movento-under practical circumstances. Established on these discoveries introductory use designs were developed and accepted to protection related useful arthropods and-to ensure-full compatibility-of the-product with-modern-IPM performs (Bielza et al., 2005; Elbert et al., 2005). Keeping in a view of the significance of entomopathogenic fungi and noval chemistry pesticides the current study was conducted to assess the evaluation of B. bassiana and M. anisopliae in integration with the insecticide against dusky cotton bugs.
- LITERATURE REVIEW
Sanjaya and Tomson (2016) evaluated that Tetranychus kanzawai is a monetarily significant polyphagous parasite species in East and South Asia. In the Philippines, it normally swarms cassava and papaya plants. The vermin assault and seriously harm the more seasoned leaves of papaya trees just as seedlings. Its genuine harm makes the leaves evaporate, in this way, lessening the photosynthetic movement of the plant. Three entomopathogenic parasitic segregates were tried on bugs under nursery conditions utilizing rewarded papaya seedlings following a totally randomized plan. The parasites tried were inspected under a dismembering extension to decide the causal operator and to affirm mortality. The LT50 of Metarhizium anisopliae, Beauveria bassiana and Paecilomyces lilacinus on T. kanzawai were assessed. Our outcomes show that among these entomopathogenic organisms, the Metarhizium anisopliae Ma6 disengage (LT50= 3.00 days) has probability for the control of T. kanzawai. Klieber et al. (2016) examined that the ongoing presentation and quick spread of the tomato leaf excavator Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) all through Europe, Africa and the Middle East represents an extreme danger to future development of tomato and other Solanaceae. Among the best practicable methods for viable and feasible control of this intrusive bug vermin are entomopathogenic parasites, which can either forestall a further spread of this creepy crawly to new regions or keep populace densities beneath a financial limit level. Here, we report on the viability of an economically accessible mycoinsecticide dependent on the entomopathogen Beauveria bassiana (Bals.) Vuill (Ascomycota: Hypocreales) against every one of the four larval phases of T. absoluta. In bioassays, high death rates and fundamentally diminished life span of hatchlings were evident when hatchlings had taken care of for a time of around 15 days on leaves with B. bassiana propagules present on a superficial level as an epiphyte with mortality esteems revised for change in charge mortality of 90–100%. What’s more, a second method of activity of B. bassiana against T. absoluta hatchlings was apparent in bioassays as an endophytic foundation of this parasite in rewarded tomato plants. Life span of T. absoluta L4 hatchlings was fundamentally lower in people which had benefited from surface‐sterilized tomato leaves acquired from plants rewarded 18 days before the bioassay with a B. bassiana suspension contrasted with hatchlings benefiting from control plants rewarded with water. Amended mortality arrived at values somewhere in the range of 30% and half for every single larval instar. Development of tomato plants was not repressed because of colonization by endophytic B. bassiana and a feeble foundational translocation of contagious propagules in non‐treated leaves was obvious in the measures. In like manner, entomopathogenic growths like B. bassiana express various methods of activity again target creepy crawly bothers, which is of specific significance for the plan of effective administration techniques for obtrusive vermin like T. absoluta. Ullah et al. (2016) evaluated that the Oxycarenus hyalinipennis Costa (Lygaidae: Hemiptera), is polyphagous in nature and has become one of the extreme sucking bugs of cotton in Pakistan.O. hyalinipennis has the possibility to develop resistance to various bug sprays, and as a result,O. hyalinipennis out breaks happen. There is no previous study from Pakistan regarding O. hyalinipennis resistance to bug sprays. In this manner, the point of this investigation was to survey the obstruction of various field populaces of O. hyalinipennis to customary (bifenthrin, deltameth-rin, lambda-cyhalothrin, profenofos, triazophos) and novel science (emamectin benzoate, spinosad, chlorfe-napyr, imidacloprid, and nitenpyram) bug sprays. Five populaces of O. hyalinipennis, gathered from Multan, Khanewal, Muzaffargarh, Lodhran, and Bahawalpur, were tried for protection from chose bug sprays by the leaf plunge technique. For three pyrethroids, the opposition proportions were in the scope of 14-to 30- overlays for bifenthrin, 2.14-to 8.41-overlap for deltamethrin, and 9.12-to 16-crease for lambda-cyhalothrin, contrasted and the laboratory susceptible strain (Lab-PK). For two organophosphates, the scope of obstruction proportions was 12-to 14-crease for master fenofos and 9.04-to 15- overlays for triazophos. For five novel science bug sprays, the scope of obstruction ratios was 4.68-to 9.83-overlap for emamectin benzoate, 6.38-to 17-overlay for spinosad, 16-to 46-crease for chlorfenapyr, 11-to 22-overlay for imidacloprid, and 1.32-to 11-overlap for nitenpyram. Customary evaluation of protection from insecticide sand coordinated administration plans likewise utilization of bug sprays and pivot of bug sprays along with different methods of activity are required to defer opposition advancement in O. hyalinipennis. Ullah et al. (2017) examined that the poisonousness of the insecticides and constancy in the dusky cotton bug field. Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae) has become a genuine financial nuisance of the malvaceae family in Pakistan. It is broadly appropriated on the planet and is polyphagous. In this examination, we researched the poisonousness of bug sprays, bug spray opposition and its dependability and cross-obstruction in abroad strain of O. hyalinipennis. Experiments of all the insecticides were done with leaf dip method. Chlorfenapyr toxicity was considerably greater than fi-pronil, bifenthrin and emamectin benzoate. Chlorfenapyr toxicity was greater as compared to the bifenthrin although chlorfenapyr toxicity was considerably low than the triazophos. Stability outcomes revealed resistance to chlorfenapyr and triazophos were not a constant whereas constant resistance was determining to the bifenthrin, fipronil and emamectin benzoate. Khan et al. (2017) were conducted an experiment to detect the periodic population of the DCB, at the NIAB, Faisalabad. The Data shown that the prevalence, of the Dusky Cotton Bug was started in second term of the September also extended to highest level in the 1st week of October. In first week of the November population, start to fall when the crop season terminate. The humidity and optimal temperature for the cotton strainer was noted on the 41% and 35.5 correspondingly. The study of the area bioassay shown a mean of the population of cotton strainers stayed at the 1.8 insect/boll in imidacloprid. Naik et al. (2017) investigated that various bug sprays were assessed on cotton creepy crawly bothers under high thickness planting framework (HDPS) on research plots at Central Institute for Cotton Research, Nagpur during kharif 2012-13. Viability of Buprofezin, fipronil, imidacloprid, diafenthiuron, spiromesifen, flonicamid and acephate were among the manufactured bug sprays while, biopesticides and botanicals included NSKE, Verticillium lecanii, coarse slaughter, coarse quit, bacterial plan, neem oil was tried against cotton creepy crawly bugs. Flonicamid half WG and Acephate, 75% SP were the best medicines which were essentially better than the various medicines was 2.19 Jassid sprites/3 leaves/plant and 2.59 fairies/3 leaves/plant separately. The least whitefly populace was recorded in the flonicamid half WG (0.42 whitefly/3 leaves/plant), Verticillium lecanii (0.50 whitefly/3 leaves/plant) and Spiromesifen (0.64 whitefly/3 leaves/plant) when contrasted with different medicines. Further, these bug sprays were seen as ecofriendly and safe to the regular foes. Saeed et al. (2017) assesed that the cotton leaf container, Amrasca devastans (Distant) is a key nuisance of cotton (Gossypium hirsutum L.), vegetables, and numerous different yields in a few Asian nations including Pakistan. Bug sprays are frequently utilized as the sole administration strategy for this irritation. Protection from neonicotinoids have been concentrated in a few creepy crawly bugs, however there is an absence of data on A. devastans. In this way, the point of this examination was to screen the obstruction in this nuisance to neonicotinoids. Six populaces of A. devastans gathered from Bahawalpur, Multan, Muzaffargarh, Rajanpur, Sahiwal and Vehari were assessed for protection from chose neonicotinoid bug sprays by the leaf plunge strategy. Obstruction proportions (RRs) were in the scope of 2.3–29.3 overlap for acetamiprid, 4.8–95.0 overlay for imidacloprid, and 19.1–1197.9 crease for thiamethoxam in various populaces of A. devastans contrasted with the reference Multan populace. Bug spray obstruction checking plans and incorporated opposition the executives approaches including the reasonable utilization of bug sprays with right application rate and techniques, revolution of bug sprays with various methods of activity and bio-control apparatuses ought to be actualized to forestall field control disappointments, especially in territories where A. devastans has created protection from the tried neonicotinoids. Rondot et al. (2018) investigated that the fungi are significant normal pathogens of arthropod bugs and are effectively utilized as biocontrol specialists in different harvests. Notwithstanding colonizing arthropods, proof has aggregated that some entomopathogenic parasites like Beauveria bassiana can endophytically colonize a wide cluster of plant species. Notwithstanding, just constrained data is right now accessible on the endophytic colonization of grapevines with B. bassiana and whether the organism despite everything keeps up its pathogenic propensity against bug bugs. Nursery and field tests were led to enhance endophytic foundation of the entomopathogenic organism B. bassiana in more youthful, pruned plants and develop grapevine plants in the vineyard. We utilized two diverse popularized B. bassiana strains, applied either as conidial suspensions (ATCC 74040 and GHA) or as a defined item (Naturalis, strain ATCC 74040) on grapevine leaves. The capability of endophytic B. bassiana to give assurance against putative objective nuisance bugs like the vine mealybug Planococcus ficus was evaluated in a bioassay utilizing surface disinfected leaves. Endophytic endurance of B. bassiana inside leaf tissues of seven-week-old pruned plants was apparent for at any rate 21 days after immunization, independent of the inoculum utilized. Sanchez et al. (2018) investigated that entomopathogenic parasites have generally been expected to help direct bug populaces. In any case, some hypocrealean ascomycetes, for example, Beauveria bassiana, play other, ineffectively comprehended biological jobs that may be helpful in creating novel methodologies for both expanded harvest creation and yield assurance. The essential points of this work were (a) to survey endophytic colonization of bread wheat and durum wheat plants by the applied growth B. bassiana strain EABb 04/01-Tip; (b) to inspect the effect of different B. bassiana vaccination techniques on development, yield, phytohormone levels and supplement take-up in the plants, and (c) to measure mortality of cotton leafworm (Spodoptera littoralis) hatchlings took care of with leaves from immunized plants. Three analyses including distinctive immunization strategies (viz., ‘soil treatment’, ‘seed dressing’ and ‘leaf showering’), and a fourth test to evaluate mortality in S. littoralis hatchlings took care of with leaves from endophytically-colonized plants, and were led by a totally randomized structure. Beauveria bassiana effectively settled inside, and colonized, bread wheat and durum wheat plants. The organism was, just because, re-separated from grains delivered by plants immunized utilizing the ‘seed dressing’ and ‘soil treatment’ strategies. ‘Seed dressing’ expanded grain yield by about 40%, and furthermore root length, in bread wheat contrasted and control plants. Mortality in S. littoralis hatchlings took care of with leaves from immunized plants ran from 30% utilizing the ‘seed dressing’ technique to 57% utilizing the ‘leaf showering’ strategy contrasted and 0% when taken care of the control leaves. Be that as it may, no contagious outgrowth was recognized in larval bodies. The manageability of yield creation and harvest security procedures dependent on B. bassiana in this manner relies upon the viability of the immunization strategy and on the specific host plant. Malik et al. (2018) evaluated that the cotton (Gossypium hirsutumL.) is the world’s driving fiber crop for material industry in numerous nations. Pakistan positions 4thin cotton creation around the world. Cotton is famous for being defenseless to numerous maladies. Bt cotton is fundamentally influenced by sucking bugs after an abatement in the force of pesticides. The accompanying investigation has been intended to check the impact of various dosages of pesticides. Three neonicotinoid bug sprays (nitenpyram, imidacloprid, and acetamiprid) and an equivalent number of ordinary bug sprays (profenophos, λ-cyhalothrin and bifenthrin) are utilized to be checked. Their impact is kept an eye on sucking creepy crawly bothers in the cotton field. RCBD (Randomized Complete Block Design) is utilized to organize the seven medicines containing six bug sprays and one control treatment with three replications each. The information of three sucking creepy crawly bugs, red cotton bug, thrips, and the shadowy cotton bug was recorded in the wake of splashing at the timespan, 72 hours and 7 days from each plot. Bug pervasion was greatest before shower butthere was a recognizable decrease after the utilization of the splash. Best outcomes were given by Acetamiprid against thrips following multi week of use followed by Nitenpyram and Profenophos. Against Dusky, Cotton Bug Nitenpyram was best trailed by Profenophos. While for Red Cotton Bug following multi week of utilization, Imidacloprid gave better outcomes followed by Lambda. Gonzalez-Mas et al. (2019) narrated that the entomopathogenic fungi are currently known to have an endophytic limit that instigates a wide scope of changes in the composition of plant supplements or potentially protective mixes. These progressions could impact collaborations between the plant and higher trophic levels. In this investigation, we assessed the predation/parasitism adequacy of hatchlings of the lacewing, Chrysoperla carnea, and the braconid parasitoid, Aphidius colemani, when offered aphids that had been tested by the entomopathogenic growth Beauveria bassiana. Aphids were either immunized straightforwardly with a parasitic suspension (lacewing bioassay just) or had been benefiting from melon plants endophytically colonized by B. bassiana. Our outcomes show that B. bassiana application didn’t essentially impact the quantity of aphid prey devoured by lacewings, or the time took them to expend every aphid. In a decision bioassay, C. carnea hatchlings wanted to benefit from aphids raised on B. bassiana-colonized plants contrasted and control plants. In another decision measure, the quantity of aphids parasitized by A. colemani and their sex proportion were not impacted by whether the aphids had been benefiting from B. bassiana-colonized plants or not. Our discoveries bolster the theory that endo-phytic entomopathogenic growths can be utilized in mix with other regular foes, for example, predators and parasitoids, in Integrated Pest Management programs. Sufyan et al. (2019) assessed that the biocontrol capability of Beauveria bassiana(Balsamo) Vuillemin and Bacillus thuringiensis(Berliner) was investigated under research facility conditions against maize stem borer Chilo partellus(Swinhoe) (Lepidoptera: Pyralidae). Three portion rates of B. bassiana (1× 104, 1×106and 1× 108conidia/ml) and one of B. thuringiensis (0.75μg/g) were applied alone and in mix against second and fourth larval instars of C. partellus. Larval mortality, pupation, grown-up development, myco-sister and sporulation fluctuated against various individual focuses of B. bassiana and its incorporated application with B. thuringiensis. Consequences of the investigation uncovered that joined use of most elevated focus of B. bassiana(1× 108spores/ml) and B. thuringiensis(0.75μg/g) displayed most noteworthy larval mortality both in second and fourth instars larvae of C. partellus. The mortality information likewise exhibited that second instar hatchlings were more vulnerable to the tried microbial treatments than fourth larval instars. In addition, less pupation and grown-up rise was seen in consolidated medicines of entomopathogens instead of their individual applications. Greatest mycosis and sporulation was recorded in the cadav-ers of C. partellus where B. bassiana was applied alone at portion pace of 1× 104conidia/ml. These results recommend thatcombined application of B. bassiana and the harmful protein created by B. thuringiens could be a promising ecofriendly approach for the fruitful administration of maize stem borer. Nasir et al. (2019) narrated that effectiveness of the diverse pesticides were checked against the DCB in the field conditions. To conclude in the decline of the population of pest 7 pesticides was drenched viz. fipronil, bifenthrin, triazophos, lambda cyhalothrin, chlorpyriphos, deltamethrin and clothianidin. Experiment was directed in using three replications. Chlorpyriphos was utmost active pesticide for the population control of cotton seed bug, with the decline of the 96.98 percent in the population monitored by the lambda, triazophos and cyhalothrin. Ahmed et al. (2020) examined that the dusky cotton bug, Oxycarenus hyalinipennis has showed up as a genuine creepy crawly vermin of different yields including cotton. The current investigation was done to assess the joined impact of some segregates of Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea and six manufactured bug sprays i.e., triazophos, imidacloprid, bifenthrin, nitenpyram, pyreproxifin and lambda cyhalothrin against O. hyalinipennis. The adequacy of these microbial operators could be upgraded by applying in mix with bug sprays. The consolidated treatment of separates of entomopathogenic growths and engineered bug sprays caused higher mortality of O. hyalinipennis. Also, the consolidated medicines diminished the grown-up rise, male and female life span, while expanded the nymphal term. The outcomes showed that these entomopathogenic organisms can furnish better control in blend with bug sprays and can be utilized in coordinated irritation the executives for decreasing the populace thickness of O. hyalinipennis.
- MATERIALS AND METHODS
The experiment was conducted in the Biological control lab of the Entomology University of Agriculture, Faisalabad, under controlled lab conditions at 28±2°C temperature and 60-70% RH. Adults and the nymphs of the cotton strainers were collected from different field, of University of Agriculture, Faisalabad (fig 3.1). Insects for bioassay were nurtured in the clear cage of 30×30cm keep up at the 28±2°C temperature, 60-70% relative humidity, 10L-14D hour of photoperiod and they fed on the both immature and mature cotton bolls. The culture was established until the required population was available for experiment.
Figure: 3.1: Collection of dusky cotton bug from cotton field
3.1 Lab Bioassay
Counted number of specimen per replication was treated against the diverse concentration of the entomopathogenic fungi and new chemistry pesticide movento there were ten treatments, containing the control and the each of treatment was replicate three times. Adult and 5th instar nymph of the dusky cotton bugs were individually sprayed with fungus solution and pesticide around 8-10 second (Fig.3.2). The treated insect of dusky cotton bugs were sited on soft paper to absorb the excessive wetness, shifted into the clear plastic jar and supplied with the lint. Mortality data was recorded on regular basis.
Figure 3.2: Dusky cotton bugs in boxes treated with Beauveria bassiana, Metarhizium anisopliae and in combination with pesticide
3.2 Beauveria bassiana
- bassiana (Racer) product was obtained from Agri. Life Hyderabad India and was used against dusky cotton bug. For the management of dusky cotton bug different concentration of B. bassiana was applied.
3.2.1 General Information
Beauveria bassiana was discovered in 1835 by Agostino bassi. First time it was reported from domesticated silkworm as the cause of muscardine disease. All over the world Entomopathogenic fungus (Beauveria bassiana) grow naturally in soil. Beauveria bassiana is a parasite against many arthropods whiteflies, aphids, bedbugs, thrips, beetles, weevils, grasshoppers, termites, malaria-transmitting mosquitoes and mealybugs. Insect susceptibility depends on various sorts of strains. From dead bodies of insect spores are extracted and prepared into spray able form.
3.2.2 Mode of action
Several different steps are involves to complete the life cycle of entomopathogenic fungi for is hyphae penetrates over cuticle, 4th step is fungi grow inside the body of host, 5th is occurrence of fungi over the body of insect and the final step is conidiogenesis. After the fungal pathogens attach to the body of host enzymes perform significant role.
3.2.3 Mode of Application
Two concentrations of B. bassiana 1×108 condia/ml and 1×106condia/ml. High humidity was necessary for the activity of B. bassiana. Spores mortality is causes by solar radiation. Spray in low temperature for its effective application. Shower should be effective due to small life period of fungus growth. Its life period is about 10 days and when ten days complete its consequence finished. The control of insect is difficult which bore into the plant. Application of entomopathogenic fungi is efficient for the management of pest population during their initial stages of growth. Mortality of insect depends on viability and number of spores contacting the pest.
3.3 Metarhizium anisopliae
Pacer (Metarhizium anisopliae) was obtained from Agri. Life Hyderabad, India and was used against dusky cotton bug population. Different concentration of Metarhizium anisopliae were applied to assess its effect against dusky cotton bug species.
3.3.1 General Information
The common name of Metrhizium anisopliae is pacer which grows naturally throughout the world in soils and act as a parasite to pests’ i.e. entomopathogenic in nature. It is parasitic to the wide range of Arthropods such as thrips, beetles, white flies, caterpillars, ants, bed bugs ,weevils, termites, grasshoppers, aphids, meallybugs, and also mosquitos (transmits malaria). Insect pests show variations to different types of strains. For the commercial use of Metrhizium anisopliae in order to make a specific product, strains of the fungus were collected and cultured from different infected insect pests. Through a process called biofermentation, the product was prepared. To make the fungus into a liquid form (sprayable), the conidia (spores) of Metrhizium anisopliae were extracted .The concentration of substainable spores of Metrhizium anisopliae in the liquid spray should be 2.5× 10. The action and infection of the fungal spores (conidia) become increased with high rate of humidity and water. The solar rays have adverse effect on the action of fungal spores.
3.3.2 Mode of Action
When the insect pests come into contact with the spores (conidia) of Metarhizium anisopliae, it kills them. The contact between the fungus and the insect pest is made by different methods. The most operative and common method is that the droplets of the fungus which are in spray from falls the insect pest or the pest itself walks to the surface of plan treated by the fungus. As the spore of fungus (conidia) adhere to the cuticle of the pest, germination of fungal spores starts and the protruded hyphae of the fungus penetrates into the body of pest and then spreads to the whole body of the insect. Death of the insect pest infected with fungus occurs within three to five days. Second proliferation of the fungus occurs as the dead insect pest play role as a source of spread. The male infected adult insect also plays role in the transmission of fungus while mating.
3.3.3 Mode of Application
The concentration of substainable spores of Metrhizium anisopliae in the liquid spray should be 2.5× 10. The solar rays have adverse effect on the action of fungal spores. So, the plant infected with insect pests should be sprayed in the dawn and dusk or in cool and moderate temperature (Butt and Goettel et al., 2000). The application of the fungus (Metarhizium anisopliae) should be done on that parts of the the plant where pests frequently occur such as on the undersides of the leaves and on the top of leaves. The fungus should be applied in a way that all parts of the plant containing pests must be covered. The life cycle of the spores is comparatively short, so the spray should be enough to contact properly with the insect pest. Control of plant borer insect is quite difficult. For these plant borers, fungus should be applied at the early growth stages of the insect in order to avoid much damage. The efficacy of the fungus to cause mortality of the insect pests depends upon environmental conditions, vulnerability of the insect pest, age of the pest and number of conidia to which the insect pest makes contact properly
3.4 Statistical analysis
The collected data was subjected to analysis of variance (ANOVA) and treatment means with significant differences will be separated using LSD test at p ≤ 0.05(Gomez and Gomez, 1984; kuchl, 2000). All statistical analysis was carried out using statistics, 8.1 packages.
- FINDINGS
4.1. Nymph mortality of 5th instar after after day 1
Significant differences (p ≤ 0.05) were detected in the 5th instar nymph mortality of Oxycarenus hyalinipennis when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis (Table 4.1). The results of the experiment revealed that maximum 5th instar nymph mortality of Oxycarenus hyalinipennis (86.67%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (78.33%). Similarly the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (75%) were also better than combined treatment of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (70%) regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (73.33%). Moreover, the individual applications of both fungal concentration showed minimal mortality and were statistically at par with each other. Overall, results showed that mortality response of Oxycarenus hyalinipennis 5th instar nymph was more profound towards combined applications of treatments rather than their individual applications (Figure 4.1).
Table 4.1: Analysis of variance table for 5th instar nymphal mortality of Oxycarenushyalinipennis after day 1 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 39536.67 | 4392.96 | 351.44 | < 0.001 |
Error | 20 | 250 | 12.5 | ||
Total | 29 | 39786.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.2. Nymph mortality of 5th instar after after day 2
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on 5th instar nymph mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis (Table 4.2). The results of the experiment revealed that maximum 5th instar nymph mortality of Oxycarenus hyalinipennis (90%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (81.67%). Similarly the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (76.67%) were also better than combined treatment of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (75%) regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (78.33%). The results further revealed that individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (26.67%) was significantly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (20%) regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis. However, besides untreated check, minimum mortality was recorded in case of individual application of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) (11.67%).
Table 4.2: Analysis of variance table for 5th instar nymphal mortality of Oxycarenushyalinipennis after day 2 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 32574.17 | 3619.35 | 241.29 | < 0.001 |
Error | 20 | 300 | 15 | ||
Total | 29 | 32874.17 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.3. Nymph mortality of 5th instar after day 3
Significant differences (p ≤ 0.05) were detected in the 5th instar nymph mortality of Oxycarenus hyalinipennis when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis (Table 4.3). The results of the experiment revealed that maximum 5th instar nymph mortality of Oxycarenus hyalinipennis (93.33%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (86.67%). Similarly the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (81.67%) were also better than combined treatment of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (80%) regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (86.67%). The results further revealed that individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (33.33%) was slightly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (31.67%) regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis.
Table 4.3: Analysis of variance table for 5th instar nymphal mortality of Oxycarenushyalinipennis after day 3 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 31703.33 | 3522.59 | 301.94 | < 0.001 |
Error | 20 | 233.33 | 11.67 | ||
Total | 29 | 31936.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.4. Nymph mortality of 5th instar after day 4
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on 5th instar nymph mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the nymphal mortality of Oxycarenus hyalinipennis (Table 4.4). The results of the experiment revealed that maximum 5th instar nymph mortality of Oxycarenus hyalinipennis (98.33%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (91.67%). However, the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (88.33%) were slightly better but statistically at par with combined treatment of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (86.67%) regarding the nymphal mortality of Oxycarenus hyalinipennis. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (91.67%) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (83.33%). However, besides untreated check, minimum mortality was recorded in case of individual application of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) (63.33%) which was statistically at par with lower dose of B. bassiana (BB1: 1×106 conidia/ml) (66.67%) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (83.33%).
Table 4.4: Analysis of variance table for nymph 5th instar mortality of Oxycarenus hyalinipennis after day 4 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 20480 | 2275.56 | 143.72 | < 0.001 |
Error | 20 | 316.67 | 15.83 | ||
Total | 29 | 20796.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.5. Nymph mortality of 5th Intsar after day 5
Significant differences (p ≤ 0.05) were detected in the 5th instar nymph mortality of Oxycarenus hyalinipennis laetus when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the nymphal mortality of Oxycarenus hyalinipennis (Table 4.5). The results of the experiment revealed that maximum 5th instar mortality of Oxycarenus hyalinipennis (100%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (96.67%). However, the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (91.67%) were statistically at par with lower dose of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (91.67%) regarding the 5th instar mortality of Oxycarenus hyalinipennis. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (96.67%). The results further revealed that individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (85%) was significantly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (75%) regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis.
Table 4.5: Analysis of variance table for 5th instar nymph mortality of Oxycarenushyalinipennis after day 5 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 21604.17 | 2400.46 | 205.75 | < 0.001 |
Error | 20 | 233.33 | 11.67 | ||
Total | 29 | 21837.5 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.6. Nymph mortality of 5th instar after day 6
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on 5th instar nymph mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the 5th instar nymph mortality of Oxycarenus hyalinipennis (Table 4.6). The results of the experiment revealed that in case of integrated application of fungal treatments maximum 5th instar nymph mortality of Oxycarenus hyalinipennis was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) (100%) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (98.33%). However, the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (96.67%) were slightly better but statistically at par with combined treatment of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (93.33%) regarding the nymphal mortality of Oxycarenus hyalinipennis. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (100%) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (86.67%).
Table 4.6: Analysis of variance table for 5th instar nymphal mortality of Oxycarenushyalinipennis after day 6 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 22550.83 | 2505.65 | 187.92 | < 0.001 |
Error | 20 | 266.67 | 13.33 | ||
Total | 29 | 22817.5 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.7. Nymph mortality of 5th instar after day 7
Significant differences (p ≤ 0.05) were detected in the 5th instar nymph mortality of Oxycarenus hyalinipennis when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the nymphal mortality of Oxycarenus hyalinipennis (Table 4.7). The results of the experiment revealed that maximum 5th instar nymph mortality of Oxycarenus hyalinipennis (100%) was observed in case of integrated application of highest and lowest concentration of B. bassiana (BB2: 1×108 conidia/ml and BB1: 1×106 conidia/ml) and highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) with Movento (1%). However, the combined applications of lower dose rate of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (98.33%) showed slightly lower but statistically same 5th instar nymph mortality of Oxycarenus hyalinipennis as compared to above mentioned treatments. However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (100%) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (90%). The results further revealed that individual applications of lower concentration of B. bassiana (BB2: 1×106 conidia/ml) (75%) was statistically at par with individual lower concentration of M. anisopliae (MA2: 1×106 conidia/ml) (70%) regarding the nymphal mortality of Oxycarenus hyalinipennis.
Table 4.7: Analysis of variance table for 5th instar nymphal mortality of Oxycarenushyalinipennis after day 7 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 22486.67 | 2498.52 | 249.85 | < 0.001 |
Error | 20 | 200 | 10 | ||
Total | 29 | 22686.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.8. Adult mortality after day 1
Significant differences (p ≤ 0.05) were detected in the adult mortality of Oxycarenus hyalinipennis when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.8). Moreover, a significantly higher adult mortality of Oxycarenus hyalinipennis was recorded in the combined applications of treatments compared to their individual applications (Figure 4.3). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis (73.33%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of lower concentration of M. anisopliae (BA1: 1×106 conidia/ml) and Movento (1%) (70%). The results further revealed that combined applications of lower dose rate of M. anisopliae (MA1: 1×106 conidia/ml) and Movento (1%) (65%) showed slightly lower but statistically same adult mortality of Oxycarenus hyalinipennis as compared to above mentioned treatments. Moreover, the individual applications of both fungal concentration showed minimal mortality and were statistically at par with each other. Overall, results showed that mortality response of Oxycarenus hyalinipennis adults was more profound towards combined applications of treatments rather than their individual applications (Figure 4.3).
Table 4.8: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 1 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 34820 | 3868.89 | 357.13 | < 0.001 |
Error | 20 | 216.67 | 10.83 | ||
Total | 29 | 35036.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
Figure 4.3: Mean effect of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) on adult mortality of Oxycarenus hyalinipennis after day 1. Bars with different lower case letters are significantly different. LSD test at p ≤ 0.05. Vertical bars indicate SE. Where; BB1: 1×106 conidia/ml; BB2: 1×108 conidia/ml; MA1: 1×106 conidia/ml; MA2: 1×108 conidia/ml; Mov: Movento: (1%) (240 SC).
4.9. Adult mortality after day 2
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on adult mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.9). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis (78.33%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (73.33%) along with integration of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (73.33%). However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (76.67%). The results further revealed that besides untreated check, minimum mortality was recorded in case of individual application of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) (13.33%) (Figure 4.4).
Table 4.9: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 2 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 26380 | 2931.11 | 219.83 | < 0.001 |
Error | 20 | 266.67 | 13.33 | ||
Total | 29 | 26646.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
Figure 4.4: Mean effect of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) on adult mortality of Oxycarenus hyalinipennis after day 2. Bars with different lower case letters are significantly different. LSD test at p ≤ 0.05. Vertical bars indicate SE. Where; BB1: 1×106 conidia/ml; BB2: 1×108 conidia/ml; MA1: 1×106 conidia/ml; MA2: 1×108 conidia/ml; Mov: Movento: (1%) (240 SC).
4.10. Adult mortality after day 3
Significant differences (p ≤ 0.05) were detected in the adult mortality of Oxycarenus hyalinipennis when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.10). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis (83.33%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by individual application Movento (1%) (80%). Furthermore, the results regarding the adult mortality of Oxycarenus hyalinipennis showed that the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (76.67%) were statistically at par with the integrated application of highest dose of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (76.67%). The results further revealed that individual applications of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) (33.33%) was slightly better but statistically at par with the individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (28.33%) regarding the adult mortality of Oxycarenus hyalinipennis.
Table 4.10: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 3 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 24920 | 2768.89 | 207.67 | < 0.001 |
Error | 20 | 266.67 | 13.33 | ||
Total | 29 | 25186.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.11. Adult mortality after day 4
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on adult mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.11). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis (88.33%) was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by individual application Movento (1%) (85%). Furthermore, the results regarding the adult mortality of Oxycarenus hyalinipennis showed that the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (81.67%) were statistically at par with the integrated application of highest dose of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (81.67%). The results further revealed that individual applications of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) (83.33%) was significantly better than the individual application of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (75%) regarding the adult mortality of Oxycarenus hyalinipennis.
Table 4.11: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 4 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 16630 | 1847.78 | 123.19 | < 0.001 |
Error | 20 | 300 | 15 | ||
Total | 29 | 16930 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.12. Adult mortality after day 5
Significant differences (p ≤ 0.05) were detected in the adult mortality of Oxycarenus hyalinipennis when tested against different individual and combined concentrations of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov). All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.12). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) (93.33%) followed by individual application Movento (1%) (90%). Furthermore, the results regarding the adult mortality of Oxycarenus hyalinipennis showed that the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (85%) were statistically at par with the integrated application of highest dose of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (86.67%). The results further revealed that individual applications of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) (88.33%) was significantly better than the individual application of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (76.67%) regarding the adult mortality of Oxycarenus hyalinipennis.
Table 4.12: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 5 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 17953.33 | 1994.81 | 170.98 | < 0.001 |
Error | 20 | 233.33 | 11.67 | ||
Total | 29 | 18186.67 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.13. Adult mortality after day 6
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on adult mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.13). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) (100%) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (96.67%). Furthermore, the results regarding the adult mortality of Oxycarenus hyalinipennis showed that the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (91.67%) were statistically at par with the integrated application of lower dose of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (90%). However, in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (95%) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (93.33%).
Table 4.13: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 6 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 20963.333 | 2329.259 | 254.1 | < 0.001 |
Error | 20 | 183.333 | 9.167 | ||
Total | 29 | 21146.667 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
4.14. Adult mortality after day 7
The individual and integrated application of Beauveria bassiana (BB), Metarhizium anisopliae (MA) and Movento (Mov) had a significant effect (p ≤ 0.05) on adult mortality of Oxycarenus hyalinipennis. All the tested treatments differed significantly (p ≤ 0.05) from untreated control regarding the adult mortality of Oxycarenus hyalinipennis (Table 4.14). The results of the experiment revealed that maximum adult mortality of Oxycarenus hyalinipennis was observed in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) (100%) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (98.33%) and individual application of Movento (1%) (98.33%). Furthermore, the results regarding the adult mortality of Oxycarenus hyalinipennis showed that the combined applications of lower dose rate of B. bassiana (BB1: 1×106 conidia/ml) and Movento (1%) (96.67%) were slightly better but statistically at par with the integrated application of lower dose of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (95%). However, in case of individual applications of fungal treatments, maximum mortality was recorded in case of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (96.67%) followed by the highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) (85%).
Table 4.14: Analysis of variance table for adult mortality of Oxycarenushyalinipennis after day 7 of treatments application
SOV | DF | SS | MS | VR | P-Value |
Treatment | 9 | 21370 | 2374.44 | 189.96 | < 0.001 |
Error | 20 | 250 | 12.5 | ||
Total | 29 | 21620 |
SOV = Source of variation; DF = Degree of freedom; SS= Sum of Squares; MS= Mean Squares; VR= Variance Ratio; p ≤ 0.001= highly significant
- DISCUSSIONS
There is a great interest all over the world for utilizing and manipulating entomopathogenic fungi for biological control of insects. Entomopathogenic fungi are just like parasite which kill or disable the insects. Unlike other pathogens like bacteria and viruses which need to be ingested for their action these fungi require contact with cuticle under favorable temperature and humidity (Dhaliwal and Koul, 2007). Entomogenous fungi have been extensively used for the control of agricultural and forest pests (Ferron, 1981; Anderson et al., 1988; Maniania, 1993).The insect pathogenic fungi have been reported to control number of important pests like Aedes aegypti(Darbro et al., 2011), Plutella xylostella, S. exigua(Freed et al., 2012a, b),Maize army worm, Mythimna separata(Malik et al., 2013) different aphid species (Akmal et al., 2013)and on stored-grain beetles (Lord, 2001). Against the cotton strainer and the red bug of cotton entomopathogenic fungi, Metarhizium anisopliae and Beauveria bassiana are used. Beauveria bassiana is well recognized microbial entomopathogen of a diverse range of arthropod species. (Jackson et al 2010). Metarhizium anisopliae is a well-characterized entomopathogen (Santi et al.2010). Entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) Sorokin (Deuteromycotina: Hyphomycetalis) is a rather common agent causing infection in natural insect populations (Ogarkov and Ogarkova, 2000; Borisov et al., 2001; Hughes et al., 2004). The tetramic acid derivative spirotetramat (brand name Movento®), has shown an outstanding performance against sucking insect pests in laboratory and greenhouse assays as well as in semi-field and field trials. The present research was carried out to assess the efficacy of Beauveria bassiana, Metarhizium anisopliae integrated with pesticide Movento against dusky cotton bug. Two different concentrations of fungus were used (1×106 condia/ml and 1×108 condia/ml) with Movento (1%) (Mov: 240 SC). According to the results or research work Beauveria bassiana 1×108 condia/ml with Movento (1%) (Mov: 240 SC) have highest percentage mortality of dusky cotton bug followed by concentration B. bassiana 1×106 condia/ml with Movento (1%) (Mov: 240 SC) and then Metarhizium anisopliae 1×108 condia/ml with Movento (1%) (Mov: 240 SC) then Metarhizium anisopliae 1×106 condia/ml with Movento (1%) (Mov: 240 SC) and the least mortality (%) was recorded on the control treatmemt. The results further revealed that individual applications of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) was significantly better than the individual application of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) regarding the mortality of Oxycarenus laetus. The minimum mortality was recorded in case of individual application of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) which was statistically at par with lower dose of B. bassiana (BB1: 1×106 conidia/ml). The result shows that in the current study the maximum mortality of adults and 5th instar nymph of dusky cotton bug after the 5,6 and 7 day of applied B. bassiana (BB2: 1×108 conidia/ml) with Movento (1%) (Mov: 240 SC). It conforms the finding of Ahmed et al. (2020) that the mixtures of fungi and insecticides at higher doses showed higher mortality than alone application. Highest percent mortality i.e., 90.0 was caused by the mixture of isolate Bb-01 and bifenthrin. Identical results were reported by Dayakar et al. (2000) when combined mixture of B. bassiana and insecticide was applied to Spodoptera litura (Fabricius). B. bassiana is highly compatible with avermectin and pyrethroids than any other insecticide (De Olivera and Neves, 2004). Previous studies focused on the potential use of combination of insecticides and fungi (Sharififard et al., 2011). However recent study revealed that in case of individual applications of tested treatments, maximum mortality was recorded in case of Movento (1%) (96.67%). Similar to present research Bruck et al. (2009) has revealed that tetramic acid.derivative-spirotetramat-(brand name Movento), an exceptional performance against the sucking insect-pests in the laboratory-and greenhouse-assays-as-well-as in the semi-field-and field-trials. There is-no cross-resistance-to any-other-insecticide. After the foliar application of spirotetramat pierces through the-leaf cuticle-and is translocated-as a spirotetramat-enol through the xylem-and-phloem, up to the developing shoots-and depressed to the roots. Current study results showes that after 4 day entomopathogenic fungi cause mortality of adults and 5th nymph instar of dusky cotton bug. The results further revealed that after 4 day individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) 83.33 %( 5th instar), 83 %( adults) was significantly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) 70 % (5th instar), 75 % (adults) mortality of adults and 5th instar nymphal of Oxycarenus hyalinipennis. Similar to the present research khan et al. (2014) used B. bassiana, M. anisopliae and I. fumosorosea against red cotton bug and dusky cotton bug. These fungi were found highly virulent when tested against D. koenigii and O. hyalinipennis. Among the fungi tested B. bassiana isolate (Bb-08) showed the highest percent mortality as compared to (I.f-2.3) and (M.a-2.3). Similar to the present research B. bassiana isolates of diverse origins proved to be highly pathogenic to the whitefly nymphs and the median lethal doses of four of the thirteen B. bassiana isolates ranged between 50 and 150 conidia/mm2. Various isolates of insect pathogenic fungi signifying six species including B. bassiana, M. anisopliae, I. fumosorosea, Isaria farinosa, Isaria flavovirescens, and Lecanicillium spp. were screened as potential biological control agents of A. aegypti. Out of these, two isolates of B. bassiana appeared to be the most promising for the control of A. aegypti (Darbro et al., 2011). After 5 day entomopathogenic fungi cause mortality of adults and 5th instar nymph of dusky cotton bug. The results further revealed that after 5 day individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) 85 %( 5th instar), 88.33 %( adults) was significantly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) 75 %( 5th instar), 76.67 %(adults) mortality of adults and 5th instar nymphal of Oxycarenus hyalinipennis. After 6 day entomopathogenic fungi cause mortality of adults and 5th nymph instar of dusky cotton bug. The results further revealed that after 5 day individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) 86.67 %( 5th instar), 93.33 %( adults) was significantly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) 77 %( 5th instar), 78 %( adults) mortality of adults and 5th nymph instar of Oxycarenus hyalinipennis. After 7 day entomopathogenic fungi cause mortality of adults and 5th instar nymph of dusky cotton bug. The results further revealed that after 5 day individual applications of highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) 90 %( 5th instar), 96.67 %( adults) was significantly better than individual highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) 80 %( 5th instar), 85 %( adults) mortality of adults and 5th instar nymph of Oxycarenus hyalinipennis.The current study demonstrates the excellent bio-control potential of B. bassiana towards the emerging threats. Minimum mortality was recorded after 4 day in case of individual application of lower dose of M. anisopliae (MA1: 1×106 conidia/ml) 63.33 % (5th instar), 70 %( adults) which was statistically at par with lower dose of B. bassiana (BB1: 1×106 conidia/ml) 66.67 %( 5th instar), 71.67 %( adults). Contrary to present research Lubeck et al. (2008) reported for the first time the use of M. anisopliae as a potential biopesticide against D. peruvianusin which females were more sensitive to fungal infection than males. Moreover, the entomopathogenic fungi showed extreme virulence against the caterpillars of Spodoptera litura (Fabricius). After 5 day B. bassiana (BB1: 1×106 conidia/ml) caused 67% mortality as lower concentration of M. anisopliae (MA1: 1×106 conidia/ml) record 75% (5th instar), 71.67% (adults) mortality of Oxycarenus hyalinipennis. After 6 day B. bassiana (BB1: 1×106 conidia/ml) caused 71.67% (5th instar), 80% (adults) mortality as lower concentration of M. anisopliae (MA1: 1×106 conidia/ml) record 66.67% (5th instar), 73.33% (adults) mortality of Oxycarenus hyalinipennis. After 7 day B. bassiana (BB1: 1×106 conidia/ml) caused 75% mortality as lower concentration of M. anisopliae (MA1: 1×106 conidia/ml) record 70% (5th instar), 78.3% (adults) mortality of Oxycarenus hyalinipennis.
- CONCLUSION
In the present study, evaluation of Metarhizium anisopliae and Beauveria bassiana in integration with the pesticide against dusky cotton bugs was assessed. Adults of the dusky cotton bugs were collected from different field of cotton of the University of Agriculture Faisalabad. Collected specimens were reared in the Biological Control Lab, Department of Entomology University of Agriculture Faisalabad under controlled lab conditions. The efficacy of these microbial agents could be enhanced by applying in combination with insecticides. Two concentrations i.e., 1×106, 1×108 conidia/ml of each entomopathogenic fungi were used in combination with insecticides against 5th instar nymphs of O. hyalinipennis. Complete randomized design (CRD) was used with ten treatment and three replications and each treatment applied homogenously in all the boxes simultaneously. Data collection was started after the application of treatments. The maximum mortality in 5th instar nymph of Oxycarenus hyalinipennis (98.33%) was observed on day 4 in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) followed by highest concentrations of B. bassiana (BB2: 1×108 conidia/ml) (83.33%). And the maximum adult mortality of Oxycarenus hyalinipennis was observed on day 6 in case of integrated application of highest concentration of B. bassiana (BB2: 1×108 conidia/ml) and Movento (1%) (Mov: 240 SC) (100%) followed by the integration of highest concentration of M. anisopliae (MA2: 1×108 conidia/ml) and Movento (1%) (96.67%). The combined treatment of entomopathogenic fungi and synthetic insecticides caused higher mortality of O. hyalinipennis.. There was no significant population reduction recorded in control conditions. The results indicated that these entomopathogenic fungi can provide better control in combination with insecticides and can be used in integrated pest management for reducing the population density of O. hyalinipennis.
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Publication History
Submitted: October 08, 2023
Accepted: October 11, 2023
Published: December 11, 2023
Identification
D-0170
Citation
Lubna Rasheed, Itrat Hussainy Khan, Nimra, Rooha Farooq, Aqsa Mubeen & Kinza Imtiaz (2023). Evaluation of Beauveria Bassiana and Metarhizium Anisopliae in Integration with Insecticide against Dusky Cotton Bug. Dinkum Journal of Natural & Scientific Innovations, 2(12):765-793.
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