Ethyl Formate Fumigation against Pineapple Mealybug, Dysmicoccus brevipes, a Quarantine Insect Pest of Pineapples.

Pineapple mealybug, Dysmicoccus brevipes (Hemiptera: Pseudococcidae), is a significant pest in pineapple production and a key trade barrier. We explored the potential use of ethyl formate (EF) as a methyl bromide alternative for the postharvest fumigation of D. brevipes in imported pineapples. When treated at 8 °C for 4 h, EF fumigation was effective against D. brevipes with LCt99, the lethal concentration × time product of EF necessary to achieve 99% mortality of D. brevipes nymphs and adults at 64.2 and 134.8 g h/m3, respectively. Sorption trials conducted with 70 g/m3 EF for 4 h at 8 °C using 7.5, 15 and 30% pineapple loading ratios (w/v) indicated that loading ratio lower than 30% is necessary to achieve the LCt99 values required to control D. brevipes. In a scaled up trial using 1 m3 chamber, EF fumigation with 70 g/m3 for 4 h at 8 °C with 20% pineapple loading ratio (w/v) resulted in a complete control of D. brevipes treated. There were no significant differences in hue values, sugar contents, firmness, and weight loss between EF-treated and untreated pineapples. Our results suggest that EF is a promising alternative to methyl bromide fumigation for the postharvest phytosanitary disinfection of D. brevipes in pineapples.

Development of a Specific Nested PCR Assay for the Detection of 16SrI Group Phytoplasmas Associated with Sisal Purple Leafroll Disease in Sisal Plants and Mealybugs.

Sisal purple leafroll disease (SPLD) is currently the most destructive disease affecting sisal in China, yet its aetiology remains unclear. In our previous research, it was verified to be associated with phytoplasmas, and nested PCR based on the 16S rRNA gene using universal primers R16mF2/R16mR1 followed by R16F2n/R16R2 was confirmed as the most effective molecular method for the detection of phytoplasmas associated with SPLD (SPLDaP). However, the method has a shortcoming of inaccuracy, for it could produce false positive results. To further manage the disease, accurate detection is needed. In this study, we developed a specific nested PCR assay using universal primers R16F2n/R16R2, followed by a set of primers designed on 16Sr gene sequences amplified from SPLDaP, nontarget bacteria from sisal plants, and other phytoplasma subgroups or groups. This established method is accurate, specific, and effective for detection of 16SrI group phytoplasma in sisal, and its sensitivity is up to 10 fg/µL of total DNA. It also minimized the false positive problem of nested PCR using universal primers R16mF2/R16mR1 followed by R16F2n/R16R2. This method was further used to verify the presence of phytoplasma in Dysmicoccusneobrevipes, and the results showed that D. neobrevipes could be infected by SPLDaP and thus could be a candidate for vector transmission assays.

Comparative proteomic analysis reveals insights into the response of Cryptolaemus montrouzieri to bottom-up transfer of cadmium and lead across a multi-trophic food chain.

Contamination of agro-ecosystems with heavy metals can affect the development and reproduction of insect natural enemies. This study reports a detailed Tandem Mass Tag based quantitative proteomic analysis of underlying mechanisms responsible for stress response of Cryptolaemus montrouzieri against heavy metals (cadmium (Cd) and lead (Pb)) transported across a multi-trophic food chain. A total of 6639 proteins were detected under Cd as well as Pb stress. In Pb versus the control cluster, 69 proteins (28 up-regulated and 41 down-regulated) were differentially expressed whereas 268 proteins were differentially expressed under Cd versus the control cluster, having 198 proteins up-regulated and 70 down-regulated proteins. The analysis of differentially expressed proteins showed that 27 proteins overlapped in both clusters representing the core proteome to Pb and Cd stress. The bioinformatics analysis demonstrated that these proteins were mapped to 57 and 99 pathways in Pb versus control and Cd versus control clusters, respectively. The functional classification by COG, GO and KEGG databases showed significant changes in protein expression by C. montrouzieri under Pb and Cd stress. The heavy metal stress (Pb and Cd) induced significant changes in expression of proteins like hexokinase (HK), succinyl-CoA, trypsin like proteins, cysteine proteases, cell division cycle proteins, and yellow gene proteins. The results provide detailed information on the protein expression levels of C. montrouzieri and will serve as basic information for future proteomic studies on heavy metal responses of insect predators within a multi-trophic food chain.

The Impact of Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) on Control of Dysmicoccus neobrevipes Beardsley (Hemiptera: Pseudococcidae).

Cryptolaemus montrouzieri (Coleoptera: Coccinellidae) is an important predator of the mealybug Dysmicoccus neobrevipes (Hemiptera: Pseudococcidae), a major pest of Agave sisalana in China. Limited reports on the efficacy of C. montrouzieri against D. neobrevipes are available. This study reports the predatory efficacy and functional response of C. montrouzieri against D. neobrevipes under laboratory conditions. The prey consumption rate per day of 4th instar larvae of C. montrouzieri feeding on 1st instar D. neobrevipes nymphs (241.3 mealybugs) was the highest among the different larval life stages of the beetle. For C. montrouzieri, the prey consumption per day of adult females (19.8 mealybugs) was significantly higher compared to males (15.2 mealybugs) when feeding on 3rd instar D. neobrevipes nymphs. The functional responses of C. montrouzieri on 1st and 2nd instar D. neobrevipes nymphs were determined as Holling type II. The search rates of C. montrouzieri 4th instar larvae towards the 1st and 2nd instar nymphs of D. neobrevipes were higher than those of the other beetle life stages. In addition, the handling times of 4th instar larvae were shorter than those of the other beetle life stages. The results from this study indicate that C. montrouzieri can be used as a predator of D. neobrevipes and, therefore, it should be evaluated further for use as a biocontrol agent in D. neobrevipes management programs.

Three new mealybug species (Hemiptera: Coccomorpha: Pseudococcidae) from Iran.

Three new mealybug species, Dysmicoccus caspianensis sp. n., Dysmicoccus zagrosicus sp. n. and Phenacoccus bromi sp. n. (Hemiptera: Pseudococcidae), collected from protected areas in Iran, are described and illustrated. The identification keys to Phenacoccus species lacking ventral multilocular disc pores and Dysmicoccus species in Iran are presented.

A survey of mealybugs infesting South-Brazilian wine vineyards

ABSTRACTMealybugs (Hemiptera: Pseudococcidae) are important pests of the grapevine Vitis spp. and are responsible for direct and indirect damage to production. The main mealybug species present in wine grapevine (Vitis vinifera L.) in Southern Brazil were identified and their incidence evaluated. Bunch-samples (n = 50) from 131 vineyards located in the Serra Gaúcha Region (RS) of Brazil were analyzed at harvest, and the occurrence of mealybugs in the roots was evaluated at the time of eradication of plants for replanting. Mealybugs were reared in laboratory until adulthood for species determination. The species Dysmicoccus brevipes (Cockerell, 1983), Dysmicoccus sp., Planococcus citri (Risso, 1813), Pl. minor(Maskell, 1897), Pseudococcus viburni (Signoret, 1875) and Pseudococcus sp. were identified in bunches. Dysmicoccus sp., D. umbambae Granara de Willink, 2009, Pl. citri and Pseudococcus sp. were found in the roots. Pl. citri (31.4%) and Dysmicoccus sp. (22.7%) were the most common species found in wine grape bunches in the Serra Gaúcha Region.

Extracellular lipase of an entomopathogenic fungus effecting larvae of a scale insect.

Lipases play an important role in the infection process of entomopathogenic fungi by hydrolyzing the ester bonds of lipoproteins, fats and waxes present on the insect surface and in the body. Here we report the purification and characterization of an extracellular lipase from Isaria fumosorosea. The enzyme was purified (138.46-fold) in three steps using (NH4 )2 SO4 precipitation followed by DEAE-cellulose and Sephadex G-100 column chromatography. The molecular weight of purified enzyme was determined to be 31 KDa by SDS-PAGE. The optimum temperature and pH for enzyme activity were 35 °C and 7.0, respectively, using p-nitrophenylpalmitate as the substrate. Lipolytic activity was enhanced in the presence of Ca(+2) , Mg(+2) , Na(+) , and NH4 (+) salts, while Zn(+2) , Fe(+2) , and Cu(+2) inhibited enzyme activity. The enzyme displayed broad substrate specificity with the highest activity observed for coconut oil and p-nitrophenyl carprate. Topical co-application of purified lipase with fungal conidial suspensions decreased the median survival time (ST50 ) of Dysmicoccus neobrevipes nymphs as compared to the fungus alone. Our results indicate that an extracellular lipase produced by I. fumosorosea can be exploited for development of enzyme-based insect management.

Sobre a nomenclatura das espécies de cochonilhas-farinhentas do cafeeiro nos Estados de Minas Gerais e Espírito Santo / About the nomenclature of coffee mealybug species in Minas Gerais and Espírito Santo States, Brazil

Root coffee (Coffea arabica L.) mealybugs (Hemiptera: Pseudococcidae) collected in Boa Esperança, southern Minas Gerais State, were identified as Dysmicoccus texensis (Tinsley) (= bispinosus Beardsley) and these from aerial part collected in Castelo, State of the Espírito Santo, as Planococcus minor (Maskell). However, Brazilian literature mentions other mealybug species of coffee tree as Pseudococcus cryptus Hempel in roots and Planococcus citri (Risso) in the aerial part. Therefore, more than one mealybug species may be occurring in the roots as in aerial part of coffee trees, and the survey and taxonomic studies are necessary before setting a control program, specially biological control.

Cochonilhas-farinhentas (Hemiptera: Pseudococcidae) das raízes do cafeeiro (Coffea arabica L.) coletadas em Boa Esperança, Estado de Minas Gerais, foram identificadas como Dysmicoccus texensis (Tinsley) (= bispinosus Beardsley) e da parte aérea dos cafeeiros em Castelo, Estado do Espírito Santo, como Planococcus minor (Maskell). Porém, na literatura brasileira são citadas outras espécies de pseudococcídeos em cafeeiros, Pseudococcus cryptus Hempel nas raízes e Planococcus citri (Risso) na parte aérea. Assim, é possível que mais de uma espécie de cochonilhas-farinhentas possam estar ocorrendo nas raízes e na parte aérea dos cafeeiros, tornando os levantamentos e estudos taxonômicos necessários para o estabelecimento de programas de controle, sobretudo o biológico.