Fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), is a destructive pest of a wide array of agricultural and horticultural crops worldwide. This in vitro research assessed the combined effect of methanolic extracts of indigenous flora of Soone Valley (Khushab, Pakistan) and nine commonly used synthetic insecticides against 3rd instar larvae of S. frugiperda using the leaf-dip bioassay method. Toxicity bioassays with twelve plant extracts revealed that the extracts of Withania somnifera (L.) Dunal, Sophora mollis (Royle) Baker and Rhazya stricta Decne. were the most effective, exhibiting minimum LC₅₀ and LT₅₀ values. Bioassays with synthetic insecticides revealed a significantly higher mortality of S. frugiperda larvae by emamectin benzoate (45%), chlorpyrifos (40%) and chlorantraniliprole (38%). Further bioassays with 10 binary combinations of these most effective botanical and synthetic insecticides showed that seven pesticidal combinations exhibited synergistic toxicity, and three combinations comprising emamectin benzoate exhibited an additive effect on the mortality of S. frugiperda larvae. GC–MS analyses of methanolic extracts of W. somnifera, S. mollis and R. stricta revealed 1,2,4-trimethyl-benzene and 3,5-dimethyl-octane, 1-ethyl-2-methyl-benzene, and 1-monolinoleoylglycerol trimethylsilyl ether, decane, and lupeol as major bioconstituents, respectively. Our results demonstrated that combining botanicals with synthetic insecticides can synergize their toxicity against S. frugiperda larvae, suggesting their potential incorporation into future IPM programs against S. frugiperda and other lepidopterous pests.

In the tropics, termites are major players in the mineralization of organic matter leading to the production of greenhouse gases including nitrous oxide (N₂O). Termites have a wide trophic diversity and their N-metabolism depends on the feeding guild. This study assessed the extent to which N₂O emission levels were determined by termite feeding guild and tested the hypothesis that termite species feeding on a diet rich in N emit higher levels of N₂O than those feeding on a diet low in N. An in-vitro incubation approach was used to determine the levels of N₂O production in 14 termite species belonging to different feeding guilds, collected from a wide range of biomes. Fungus-growing and soil-feeding termites emit N₂O. The N₂O production levels varied considerably, ranging from 13.14 to 117.62 ng N₂O-N d^-1 (g dry wt.)^-1 for soil-feeding species, with Cubitermes spp. having the highest production levels, and from 39.61 to 65.61 ng N₂O-N d^-1 (g dry wt.)^-1 for fungus-growing species. Wood-feeding termites were net N₂O consumers rather than N₂O producers with a consumption ranging from 16.09 to 45.22 ng N₂O-N d^-1 (g dry wt.)^-1. Incubating live termites together with their mound increased the levels of N₂O production by between 6 and 13 fold for soil-feeders, with the highest increase in Capritermes capricornis, and between 14 and 34 fold for fungus-growers, with the highest increase in Macrotermes muelleri. Ammonia-oxidizing (amoA-AOB and amoA-AOA) and denitrifying (nirK, nirS, nosZ) gene markers were detected in the guts of all termite species studied. No correlation was found between the abundance of these marker genes and the levels of N₂O production from different feeding guilds. Overall, these results support the hypothesis that N₂O production rates were higher in termites feeding on substrates with higher N content, such as soil and fungi, compared to those feeding on N-poor wood.

Background: As different biogeographic strains and isolates of entomopathogenic fungi vary in their genetic, enzymatic and pathogenic characteristics, this study assessed the virulence of 2 indigenous strains of Beauveria bassiana (Balsam) Vuillemin and Metarhizium anisopliae (Metschn.) Sorokin (Ascomycota, Hypocreales: Clavicipitaceae), isolated from naturally infected insect cadavers, against the 3rd instar nymphs of Myzus persicae (Sulzer) (Hemiptera: Aphididae) and 3^rd instar larvae of Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae) using leaf-dip and larval-dip methods, respectively. Results: Both fungal isolates exhibited considerable pathogenicity against M. persicae and S. frugiperda. Mortality in all bioassays was conidial concentration and exposure time dependent and increased significantly along with both factors (R² = 0.86–0.99 for B. bassiana and 0.82–0.94 for M. anisopliae). Moreover, M. anisopliae isolate appeared more virulent to S. frugiperda larvae than B. bassiana isolate, while the later fungal isolate was more lethal to M. persicae nymphs than the former one. At the highest conidial concentration (1.0 × 10⁹ conidia/ml), M. anisopliae caused maximum mean mortality of S. frugiperda (88%) and M. persicae (65%) and B. bassiana exhibited maximum mean mortality of S. frugiperda (76%) and M. persicae (94%). Moreover, probit regression analyses showed LT₅₀ values for M. persicae of 4.57 and 6.86 days at 1.0 × 10⁹ conidia/ml for the isolates of B. bassiana and M. anisopliae, respectively, while LC₅₀ values were 7.75 × 10⁶ and 8.70 × 10⁷ conidia/ml after 10th day of application, for the isolates of B. bassiana and M. anisopliae, respectively, against M. persicae. Similarly, LT₅₀ values for S. frugiperda were 7.75 and 7.03 days for 1.0 × 10⁹ conidia/ml concentration and LC₅₀ values were 2.84 × 10⁷ and 8.84 × 10⁵ conidia/ml at 10th day data for the isolates of B. bassiana and M. anisopliae, respectively. Conclusion: Overall study results demonstrated the effectiveness of B. bassiana and M. anisopliae against M. persicae and S. frugiperda, respectively. However, field evaluations of these indigenously isolated promising fungal strains against these insect pests.

Coccinellid beetles (Coccinellidae; Coleoptera) are voracious predators of many insect pests of economic importance. Among 6,000 species of coccinellids described worldwide, only 75 species have been reported from Pakistan so far. Keeping in view the limited work on these important beetles, an extensive faunal survey was conducted in the district Sargodha (32°05’02’’ N and 72°40’18’’ E), Pakistan to assess the species richness and distribution of coccinellid beetles in seven selected regions (Bhera, Bhalwal, Kot Momin, Sahiwal, Sargodha, Shahpur and Silanwali). A total of 1,470 coccinellid specimens were collected from all types of vegetation and were identified up to species level by means of external morphology, published descriptions and male genitalia micrometry. Nine species were identified belonging to 9 genera and 4 subfamilies occurring along with 10 trophic associations. Five species viz; Coccinella septempunctata (Linnaeus 1758), Hippodamia variegata (Goeze 1777), Menochilus sexmaculatus (Fabricius 1781), Micraspis allardi (Mulsant 1853) and Propylea dissecta (Mulsant 1850) belonged to tribe Coccinellini (Latreille 1807) of subfamily Coccinellinae (Latreille 1807). Two species belonged to tribe Chilocorini (Costa 1849) of subfamily Chilocorinae (Mulsant 1846) i.e. Brumoides suturalis (Fabricius 1798) and Exochomus nigripennis (Erichson 1843). Two species namely Henosepilachna vigintioctopunctata (Fabricius 1775) and Scymnus nubilus (Mulsant 1850) represented respectively the tribe Epilachnini (Mulsant 1846) of subfamily Epilachninae (Mulsant 1846) and tribe Scymnini (Mulsant 1846) of subfamily Scymninae (Mulsant 1846). Among all the species, C. septempunctata was found widely distributed in all localities studied, whereas, Sargodha locality represented highest diversity indices of Shannon-Weiner’s (1.65), Margalef’s (1.40) and Simpson’s (0.75).

Green peach aphid, Myzus persicae (Hemiptera: Aphididae), is an economically important pest of crops within more than 40 plant families all over the world. This study encompasses in-vitro pathogenicity of two strains of Beauveria bassiana (BB-72 and BB-252) and one strain of Lecanicillium lecanii (V-4) against green peach aphid (M. persicae). Using a leaf-dip method, three different bioassays were conducted comprised of filtrates and conidial concentrations of BB-72, BB-252 and V-4 fungal strains and their binary combinations. Infiltrate bioassays, 2 mL fungal filtrate of each strain was used. In conidial bioassays, three different concentrations (i.e., 1 × 10⁶, 1 × 10⁷ and 1 × 10⁸ conidia mL⁻¹) of each fungal strain were used, while in binary combination bioassays, LC₅₀ and LC₃₃ of these fungal strains were evaluated. According to the results, maximum pathogenicity against test aphids was observed at 10th day of all treatments. Both strains of B. bassiana (BB-72 and BB-252) exhibited higher mortality of aphids (up to 95 and 91%, respectively) than L. lecanii (V-4) strain (87%) in all three types of bioassays. Moreover, binary combinations of BB-72 and BB-252 strains showed significantly higher aphid mortality (94%) than other combinations. Reduced mortality in case of fungal combinations of V-4 strain might be due to the antagonistic effect of L. lecanii strain along with both strains of B. bassiana. Nevertheless, evaluation of combined pathogenicity of the fungal strains constitutes the novelty of this in-vitro study which revealed that both strains of B. bassiana (i.e., BB-72 and BB-252) are mutually compatible and can be utilized together as new biocontrol tools against destructive insect pests such as M. persicae.