Insecticidal Activity of Tannins from Selected Brown Macroalgae against the Cotton Leafhopper Amrasca devastans.

Seaweeds, also known as marine macroalgae, are renewable biological resources that are found worldwide and possess a wide variety of secondary metabolites, including tannins. Drifted brown seaweed (DBSW) is particularly rich in tannins and is regarded as biological trash. The cotton leaf hopper Amrasca devastans (Distant) has caused both quantitative and qualitative losses in cotton production. Drifted brown seaweeds (DBSWs) were used in this study to extract, qualitatively profile, and quantify the levels of total tannins, condensed tannins, hydrolyzable tannins, and phlorotannins in the seaweeds; test their insecticidal activity; and determine the mechanism of action. The largest amount of tannin extract was found in Sargassum wightii Greville (20.62%) using the Soxhlet method (SM). Significantly higher amounts of hydrolyzable tannins (p = 0.005), soluble phlorotannins (p = 0.005), total tannins in the SM (p = 0.003), and total tannins in the cold percolation method (p = 0.005) were recorded in S. wightii. However, high levels of condensed tannins (CTAs) were observed in Turbinaria ornata (Turner) J. Agardh (p = 0.004). A. devastans nymphs and adults were examined for oral toxicity (OT) and contact toxicity (CT) against DBSW tannin crude extract and column chromatographic fractions 1 (Rf = 0.86) and 2 (Rf = 0.88). Stoechospermum polypodioides (J.V. Lamouroux) J. Agardh crude tannin was highly effective against A. devastans using the OT method (LC50, 0.044%) when compared with the standard gallic acid (LC50, 0.044%) and tannic acid (LC50, 0.122%). Similarly, S. wightii fraction 2 (LC50, 0.007%) showed a greater insecticidal effect against A. devastans adults in OT than gallic acid (LC50, 0.034%) and tannic acid (LC50, 0.022%). The mechanism of action results show that A. devastans adults treated with crude tannin of T. ornata had significantly decreased amylase, protease (p = 0.005), and invertase (p = 0.003) levels when compared with the detoxification enzymes. The levels of glycosidase, lactate dehydrogenase, esterase, lipase, invertase, and acid phosphate activities (p = 0.005) of S. wightii were reduced when compared with those of the Vijayneem and chemical pesticide Monocrotophos. In adult insects treated with LC50 concentrations of S. wightii tannin fraction 1, the total body protein (9.00 µg/µL) was significantly reduced (OT, LC50-0.019%). The SDS-PAGE analysis results also show that S. wightii tannin fraction 1 (OT and CT), fraction 2 (OT), and S. polypodioides fraction 2 (CT) had a significant effect on the total body portion level, appearance, and disappearance of some proteins and polypeptides. This study shows that the selected brown macroalgae can be utilized for the safer management of cotton leaf hoppers.

Insecticidal Mechanism of Botanical Crude Extracts and Their Silver Nanoliquids on Phenacoccus solenopsis.

In recent years, intensive studies have been carried out on the management of agricultural insect pests using botanical insecticides in order to decrease the associated environmental hazards. Many studies have tested and characterized the toxic action of plant extracts. Four plant extracts (Justicia adhatoda, Ipomea carnea, Pongamia glabra, and Annona squamosa) containing silver nanoparticles (AgNPs) were studied for their effects on Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) using the leaf dip method. The effects were estimated based on assays of hydrolytic enzyme (amylase, protease, lipase, acid phosphatase, glycosidase, trehalase, phospholipase A2, and invertase) and detoxification enzyme (esterase and lactate dehydrogenase) levels; macromolecular content (total body protein, carbohydrate, and lipid); and protein profile. The results show that the total body of P. solenopsis contains trypsin, pepsin, invertase, lipase, and amylase, whereas J. adathoda and I. carnea aqueous extracts considerably decreased the protease and phospholipase A2 levels, and A. squamosa aqueous extract dramatically increased the trehalase level in a dose-dependent manner. The enzyme levels were dramatically decreased by P. glabura-AgNPs (invertase, protease, trehalase, lipase, and phospholipase A2); I. carnea-AgNPs (invertase, lipase, and phospholipase A2); A. squamosa-AgNPs (protease, phospholipase A2); and J. adathoda-AgNPs (protease, lipase, and acid phosphatase). Plant extracts and their AgNPs significantly reduced P. solenopsis esterase and lactate dehydrogenase levels in a dose-dependent manner. At higher concentrations (10%), all of the investigated plants and their AgNPs consistently decreased the total body carbohydrate, protein, and fat levels. It is clear that the plant extracts, either crude or together with AgNPs, may result in the insects having inadequate nutritional capacity, which will impact on all critical actions of the affected hydrolytic and detoxication enzymes.

A chalcone (Pongamol) and phytoconstituents of Tephrosia purpurea.

Tephrosia purpurea stem extract fractions shows more amount of Cyclohexane, 1-ethenyl-1-methyl-2,4-bis(1-methylethenyl)-, (1α,2ß,4ß)- (62.32%). Secondary metabolites contents were higher in stem than root of T. purpurea. It is the first time a chalcone (Pongamol) was crystallised from the column chromatography fraction of T. purpurea stem extract identified by using FT-IR and XRD. Quantitative analysis of T. purpurea stem and root reveals that total saponins, tannins and phenols (mg/g plant powder) contents were higher in the former than the latter part of the plant, respectively.

Biofoam of Spittlebug, Poophilus costalis (Walker): Preferential Sites, Temperature Regulation, Chemical Composition and Antimicrobial Activity.

The foam produced by nymphs of Poophilus costalis on eleven different host plants belonging to eight families on St. Xavier's College campus in India was studied over five months. The chemical composition and antimicrobial activity of these biofoams were investigated. The results revealed that P. costalis preferred Theporsia purpurea and Mimosa pudica for laying their eggs and producing foam, over the other tested plants. P. costalis produce their foam on either nodes or internodes on monocotyledons (30%) (p < 0.05), whereas on dicotyledons, they produce more foam on the stems (63.8%) than on the leaves (6.2%) (p < 0.01). The number of nymphs in each piece of foam from P. costalis varied from 1 to 3 (mean = 1.8 per plant). They produced their foam (5.7 to 45.2 cm) from the ground level on a plant. The length and breadth of a piece of foam ranged from 1.0 to 3.9 cm and 0.6 to 4.7 cm, respectively. The foam tended to be cooler than the environment. Qualitative profiling showed that the foam consists of carbohydrates, including maltose; trypsin; amino acids; protease. The foam was also analyzed using a spectrophotometer, Fourier transform infrared spectroscopy (FT-IR), gas chromatography-mass spectroscopy (GC-MS), and high-performance liquid chromatography (HPLC). The antimicrobial activity of the biofoam was the greatest against Staphylococcus aureus, the growth of which was reduced by 55.9 ± 3.9%, suggesting that the foam could be used as an antimicrobial product. However, no activities were observed against Fusarium oxysporum and Candida albicans.

Bioefficacy of Rhynocoris longifrons (Stål) (Heteroptera: Reduviidae) against multiple cotton pests under screen house and field conditions.

Rhynocoris longifrons (Hemiptera: Reduviidae) is a generalist predator of many cotton insect pests. The hiding behaviour of this predator, which is one of the key factors of predation success, was investigated under screen house conditions. Moreover, we evaluated its biocontrol potential against Aphis gossypii (Hemiptera: Aphididae), Dysdercus cingulatus (Hemiptera: Pyrrhocoridae), Phenacoccus solenopsis (Hemiptera: Pseudococcidae), and Helicoverpa armigera Hübner (Lepidoptera: Noctuidae) under screen house and field conditions. Results showed that R. longifrons life stages preferred to hide under small pebbles in the screen house tests. All the R. longifrons life stages showed a biocontrol potential against the four insect pests under screen house conditions. However, their biocontrol potential had not varied in relation to day and night hours. Augmentative releases of R. longifrons were carried out for two seasons such as South-west monsoon, 2011 and post-monsoon, 2012. The augmentative release of R. longifrons reduced significantly insect pests on cotton. In fact, the release of this predator in cotton fields was capable to reduce the population of H. armigera (50%), P. solenopsis (28%), D. cingulatus (18.8%), and A. gossypii (11.8%) during the rain fed condition (south-west monsoon season). During irrigated condition (post-monsoon season), populations of D. cingulatus were reduced by 26%, than P. solenopsis (20.6%), and A. gossypii (16.8%). Except ants, no negative impact was reported on other natural enemies present in the cotton field. Significantly higher crop yield and cost benefit ratio was observed in the predator release plots indicating that R. longifrons can be used in an integrated pest management program for multiple cotton pests.

Toxicity and developmental effect of cucurbitacin E from Citrullus colocynthis L. (Cucurbitales: Cucurbitaceae) against Spodoptera litura Fab. and a non-target earthworm Eisenia fetida Savigny.

Pest management with natural botanical insecticides is a significant implementation for the sustainability of agroecosystem by reducing the unnecessary risk from the inputs of synthetic insecticides. In this research, we isolated the bioactive compound cucurbitacin E from Citrullus colocynthis (L.) Schrad, and their toxicological effects were screened against different larval instars of Spodoptera litura. The bioactive compound cucurbitacin E was chemically characterized through TLC, FT-IR, and NMR analyses. The larval mortality bioassay revealed that the larvae exposed to cucurbitacin E at the discriminating dose of 50 ppm display higher mortality rate against second (93.8%), third (86.4%), and fourth (73.2%) instar respectively. The lethal concentrations (LC50 and LC90) was detected as 15.84 and 67.60 ppm for third instar respectively. The sub-lethal concentration of cucurbitacin E (2, 4, and 6 ppm) intentionally altered the percentage of survival, pupation, fecundity, and egg hatchability of S. litura. Moreover, antifeedant activity of cucurbitacin E was analyzed using choice-based test. In addition, we found the toxic effects of cucurbitacin E (50 and 100 ppm) and chemical pesticides (cypermethrin and monocrotophos) against terrestrial beneficial earthworm Eisenia fetida, and the result revealed that cucurbitacin E has no harmful effect on non-target organism. Hence, the present study reveals that cucurbitacin E might be a part of a new biorational product alternative to synthetic pesticides.

Green preparation of seaweed-based silver nano-liquid for cotton pathogenic fungi management.

Silver nanoparticles (Ag NPs) were synthesised using the crude ethyl acetate extracts of Ulva lactuca and evaluated their bioefficacy against two crop-damaging pathogens. The sets of lattice planes in the XRD spectrum for the Ag NPs were indexed to the 111, 200, 220 and 311 orientations and support the crystalline nature of the Ag NPs. The 3414 and 2968 cm-1 peaks were observed in crude algal thallus extract and they were characteristic of terpenoids. Further, a peak at 1389 cm-1 was observed as fatty acids. The marine macroalgae terpenoids and palmitic acid acted as reducing agent and stabiliser, respectively. The size (3 and 50 nm) and shape (spherical) of Ag NPs were recorded. The energy-dispersive X-ray spectroscopy analysis exemplified the presence of silver in its elemental nature. Moreover, U. lactuca Ag NPs were effective against two cotton phytopathogens namely Fusarium oxysporum f.sp. vasinfectum (FOV) and Xanthomonas campestris pv. malvacearum (XAM). The minimum inhibitory concentration was found to be 80.0 and 43.33 µg ml-1 against FOV and XAM, respectively. Results confirmed the anti-microbial activity of green nanoparticles against select pathogens and suggest their possible usage in developing antifungal agents for controlling destructive pathogens in a cotton agroecosystem.

Lethal and Sublethal Effects of Three Microbial Biocontrol Agents on Spodoptera litura and Its Natural Predator Rhynocoris kumarii.

Entomopathogenic microbes such as Spodoptera litura nucleopolyhedrovirus (SpltNPV), Metarhizium anisopliae, and Pseudomonas fluorescens are biological agents used for the control of multiple arthropod pests. The objective of this study was to assess their effects on the biological parameters of Spodoptera litura (Lepidoptera: Noctuidae) larvae, and its natural reduviid predator Rhynocoris kumarii (Hemiptera: Reduviidae) under laboratory conditions. Results suggested that P. fluorescens reduced the food consumption index, relative growth rate, approximate digestibility, the efficiency of conversion of ingested food, and the efficiency of conversion of digested food of S. litura third instar larvae compared to prey infected with M. anisopliae and SpltNPV. Both SpltNPV and M. anisopliae caused similar mortality of S. litura life stages after 96 h of observation. To observe the effect of an infected prey diet on predator behavior, infected S. litura larvae were offered to the third, fourth, and fifth instar nymphs of R. kumarii, and their prey handling time, predation rate (number/day/predator), developmental period, and the survival rate was recorded. When the life stages of R. kumarii were offered entomopathogen-infected S. litura larvae, their predation rate was comparable to or higher than the untreated control. The juvenile predator, after feeding on P. fluorescens-infected S. litura larvae, had a significantly longer developmental period (2⁻4 days) compared to those fed on larvae infected with other microbial control agents. However, feeding on P. fluorescens alone did not affect the predator nymphal survival rate or the adult sex ratio. Although three entomopathogens had some degree of effect on the biological parameters of R. kumarii, the outcome of this study suggests that integration of reduviids with the tested entomopathogens are a compatible and potentially effective strategy for the management of S. litura populations. However promising, this combined strategy needs to be tested under field conditions to confirm the laboratory findings.

Mass rearing and augmentative biological control evaluation of Rhynocoris fuscipes (Hemiptera: Reduviidae) against multiple pests of cotton.

BACKGROUND: Rhynocoris fuscipes (Fab.) (Hemiptera: Reduviidae) is a generalist predator of cotton pests and is commonly found inhabiting cotton-growing regions in southern India. With the goal of integrating this predator in standard management practices used against cotton pests on a commercial scale, (1) we developed a protocol for adult group rearing of this predator inside micro-environmental cages (MECs), and (2) we evaluated the biocontrol potential of mass-produced predators against cotton pests under potted and field conditions. RESULTS: Higher fecundity and adult longevity of R. fuscipes was recorded in the MECs than under natural growing conditions. The reduviid predator preferred stones and fallen leaves as hiding places in the MECs. The predator showed a higher biocontrol potential during the night hours against two pests, Phenacoccus solenopsis Tinsley and Dysdercus cingulatus (Fab.), than during the day under potted conditions. Under field conditions, R. fuscipes significantly reduced the population of Aphis gossypii Glover, P. solenopsis, D. cingulatus and Helicoverpa armigera (Hübner) by 28, 70, 29 and 50%, respectively. No negative impact of R. fuscipes was reported on other natural enemies present in the cotton agroecosystem. CONCLUSION: Significantly higher crop yield and cost benefit ratio were observed in R. fuscipes-released plots than in the control plots. The results suggest that R. fuscipes can be mass produced efficiently under controlled conditions in MECs, and used in an integrated management program for multiple cotton pests. © 2017 Society of Chemical Industry.

Route of infection and hematological effect of Metarhizium anisopliae (Metsch.) Sorokin on Dysdercus cingulatus (Fab.) adult.

The primary objective of this work was to identify, under laboratory conditions, the route of infection and hemogram of Dysdercus cingulatus (Fab.) adults by Metarhizium anisopliae. The infection process in D. cingulatus by M. anisopliae involved the conidia adherence to the host cuticle and germination after 24 h post-infection, accompanied by falling of bristles. The subsequent step, within 24-48 h post-infection, comprised penetration of fungus through spiracles, root of bristles, hemolymph, and the three dorsal sacs. Subsequently, within 72-96 h post-infection, the fungus penetrated into trachea and sacs, then emerged on cuticular surface and was found to be maximum in hemolymph. A great decrease in hemocytes count was observed within 96 h from infection. The hemosomic index (HSI) decreased gradually as the incubation period increased. As far as we know, this is the first study to know the mechanism of action of M. anisopliae to D. cingulatus.

Stage preference and functional response of Rhynocoris longifrons (Stål) (Hemiptera: Reduviidae) on three hemipteran cotton pests

In this work, the stage preference and functional response of the indigenous reduviid bug Rhynocoris longifrons feeding on five different densities of the cotton aphid Aphis gossypii, Phenacoccus solenopsis, and Dysdercus cingulatus was examined in Petri dish arenas containing cotton leaves under laboratory conditions. The reduviid predator exhibited a Type II functional response at all hemipteran pests evaluated when data were fit to Holling's disc equation. Predatory rate gradually increased while the predator grew older and adults consumed maximum number of D. cingulatus and P. solenopsis. An opposite trend was observed, while the reduviid was provided with Aphis gossypii. The rate of attack on P. solenopsis was quite low but fairly consistent, with the different life stages of the predator generally more effective. Further investigation of the biological control potential of R. longifrons against cotton pests under pot and controlled filed should be done due to the predator's ability to kill adult stages of all prey species evaluated. These results indicated that R. longifrons could eat more aphids at high prey densities; however, predators also considerably reduced other cotton pests too so it could be considered a prospective candidate for use as a commercial biological control agent for cotton hemipteran pests in India.