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Optimization method for proteomic analysis of the larva and adult tissues of plutella xylostella (l.) (lepidoptera: plutellidae)

Maizom Hassan, NorazilaYusoff, Wan Mohd Aizat, Idris Abd Ghani, Nurul Wahida Othman

Sains Malaysiana, 2018;47:2975-2983.

Plutella xylostella (L.) (Lepidoptera: Plutellidae), the major insect pest of cruciferous crops worldwide shows significant
resistance to almost all classes of insecticides. In order to effectively prevent and manage the insecticidal resistance,
it is crucial to understand the physiological adaptation of insects against insecticides. Identification of insect protein
that interacting with insecticides and characterization of their modification in resistant strains can be done by using
differential proteomics approach. This study focuses on optimizing a sensitive and rapid method for the extraction of
high quality protein of both larva and adult tissues of P. xylostella to be used in two-dimensional gel electrophoresis.
Five extraction methods were evaluated for protein concentration, yields and resolving patterns of one-dimensional
and two-dimensional electrophoresis. The results showed that trichloroacetic acid/acetone extraction methods with
two different concentrations of 2-mercaptoethanol produced the highest protein concentration and yield for both adult
and larva tissues, respectively. Meanwhile, trichloroacetic acid/acetone with dithiothreitol extraction method gave
better separation of spots and intensity for both larva and adult tissues compared to other methods tested. As such, we
concluded that trichloroacetic acid/acetone with dithiothreitol successfully yielded high total protein concentration and
good separation of two-dimensional electrophoresis gel spots in both adult and larva P. xylostella.

Matched MeSH terms: Lepidoptera
Assessing diet of the rufous-winged philentoma (philentoma pyrhoptera) in lowland tropical forest using next-generation sequencing

Mohammad Saiful Mansor, Shukor Md. Nor, Rosli Ramli

Sains Malaysiana, 2018;47:1045-1050.

Dietary study provides understanding in predator-prey relationships, yet diet of tropical forest birds is poorly understood.
In this study, a non-invasive method, next-generation sequencing (Illumina MiSeq platform) was used to identify prey in
the faecal samples of the Rufous-winged Philentoma (Philentoma pyrhoptera). Dietary samples were collected in lowland
tropical forest of central Peninsular Malaysia. A general invertebrate primer pair was used for the first time to assess
diet of tropical birds. The USEARCH was used to cluster the COI mtDNA sequences into Operational Taxonomic Unit (OTU).
OTU sequences were aligned and queried through the GenBank or Biodiversity of Life Database (BOLD). We identified
26 distinct arthropod taxa from 31 OTUs. Of all OTUs, there was three that could be identified up to species level, 20 to
genus level, three to family level and five could not assigned to any taxa (the BLAST hits were poor). All sequences were
identified to class Insecta belonging to 18 families from four orders, where Lepidoptera representing major insect order
consumed by study bird species. This non-invasive molecular approach provides a practical and rapid technique to
understand of how energy flows across ecosystems. This technique could be very useful to screen for possible particular
pest insects consumed by insectivores (e.g. birds and bats) in crop plantation. A comprehensive arthropod studies and
local reference sequences need to be added to the database to improve the proportion of sequences that can be identified.

Matched MeSH terms: Lepidoptera
Fulltext Insect community in riparian zones of Sungai Sepetang, Sungai Rembau and Sungai Chukai of Peninsular Malaysia

Abdullah NA, Radzi SNF, Asri LN, Idris NS, Husin S, Sulaiman A, et al.

Biodivers Data J, 2019;7:e35679.
PMID: 31582889 DOI: 10.3897/BDJ.7.e35679

Riparian areas hold vast number of flora and fauna with exceptional contributions to the ecosystem. A study was conducted in Sungai Sepetang, Sungai Rembau and Sungai Chukai to identify the insect community in a riparian zone of Peninsular Malaysia. Sampling was conducted in six consecutive months from December 2017 to May 2018 during both day and night using sweep nets. Twenty sampling stations (S1-S20) had been assembled along the riverbanks with an average distance of 200 m between each station. The 17,530 collected insects were from 11 orders and consisted of Diptera, Coleoptera, Hemiptera, Hymenoptera, Lepidoptera, Neuroptera, Orthoptera, Blattodea, Thysanoptera, Mantodea and Odonata. The three most abundant orders were Diptera (33.84%; 5933 individuals), Coleoptera (28.82%; 5053 individuals) and Hemiptera (25.62%: 4491 individuals). The collected insect community consisted of different guilds such as the scavenger, predator, herbivore, pollinator and parasitoid. Sungai Sepetang and Sungai Rembau were dominated by mangrove flora, Sonneratia caseolaris (Myrtales: Lythraceae), while Sungai Chukai was dominated by Barringtonia racemosa. There was a significant difference (p < 0.05) in the composition of insects between the three rivers though clustering analysis showed that the insect communities in Sungai Sepetang and Sungai Rembau were 100% similar compared to Sungai Chukai which consisted of a totally different community. There is a significant negative correlation between abundance of insects with salinity and wind speed at Sungai Chukai and Sungai Sepetang.

Matched MeSH terms: Lepidoptera
Fulltext Persistent mosquito fogging can be detrimental to non-target invertebrates in an urban tropical forest

Lee NSM, Clements GR, Ting ASY, Wong ZH, Yek SH

PeerJ, 2020;8:e10033.
PMID: 33062440 DOI: 10.7717/peerj.10033

Background: Human population growth has led to biodiversity declines in tropical cities. While habitat loss and fragmentation have been the main drivers of urban biodiversity loss, man-made interventions to reduce health risks have also emerged as an unintentional threat. For instance, insecticide fogging to control mosquito populations has become the most common method of preventing the expansion of mosquito-borne diseases such as Dengue. However, the effectiveness of fogging in killing mosquitoes has been called into question. One concern is the unintended effect of insecticide fogging on non-target invertebrates that are crucial for the maintenance of urban ecosystems. Here, we investigate the impacts of fogging on: (1) target invertebrate taxon (Diptera, including mosquitoes); (2) non-target invertebrate taxa; and (3) the foraging behavior of an invertebrate pollinator taxon (Lepidoptera) within an urban tropical forest.
Methods: We carried out fogging with Pyrethroid insecticide (Detral 2.5 EC) at 10 different sites in a forest situated in the state of Selangor, Peninsular Malaysia. Across the sites, we counted the numbers of knocked-down invertebrates and identified them based on morphology to different taxa. We constructed Bayesian hierarchical Poisson regression models to investigate the effects of fogging on: (1) a target invertebrate taxon (Diptera) 3-h post-fogging; (2) selected non-target invertebrate taxa 3-h post-fogging; and (3) an invertebrate pollinator taxon (Lepidoptera) 24-h post-fogging.
Results: A total of 1,874 invertebrates from 19 invertebrate orders were knocked down by the fogging treatment across the 10 sites. Furthermore, 72.7% of the invertebrates counted 3-h post-fogging was considered dead. Our regression models showed that given the data and prior information, the probability that fogging had a negative effect on invertebrate taxa 3-h post-fogging was 100%, with reductions to 11% of the pre-fogging count of live individuals for the target invertebrate taxon (Diptera), and between 5% and 58% of the pre-fogging count of live individuals for non-target invertebrate taxa. For the invertebrate pollinator, the probability that fogging had a negative effect 24-h post-fogging was also 100%, with reductions to 53% of the pre-fogging count of live individuals.
Discussion: Our Bayesian models unequivocally demonstrate that fogging has detrimental effects on one pollinator order and non-target invertebrate orders, especially taxa that have comparatively lower levels of chitinisation. While fogging is effective in killing the target order (Diptera), no mosquitos were found dead in our experiment. In order to maintain urban biodiversity, we recommend that health authorities and the private sector move away from persistent insecticide fogging and to explore alternative measures to control adult mosquito populations.

Matched MeSH terms: Lepidoptera
Fulltext Integrative taxonomy and analysis of species richness patterns of nocturnal Darwin wasps of the genus Enicospilus Stephens (Hymenoptera, Ichneumonidae, Ophioninae) in Japan

Shimizu S, Broad GR, Maeto K

Zookeys, 2020;990:1-144.
PMID: 33269011 DOI: 10.3897/zookeys.990.55542

The predominantly tropical ophionine genus Enicospilus Stephens, 1835 is one of the largest genera of Darwin wasps (Hymenoptera, Ichneumonidae), with more than 700 extant species worldwide that are usually crepuscular or nocturnal and are parasitoids of Lepidoptera larvae. In the present study, the Japanese species of Enicospilus are revised using an integrative approach (combined morphology and DNA barcoding). On the basis of 3,110 specimens, 47 Enicospilus species are recognised in Japan, eight of which are new species (E. acutus Shimizu, sp. nov., E. kunigamiensis Shimizu, sp. nov., E. limnophilus Shimizu, sp. nov., E. matsumurai Shimizu, sp. nov., E. pseudopuncticulatus Shimizu, sp. nov., E. sharkeyi Shimizu, sp. nov., E. takakuwai Shimizu, sp. nov., and E. unctus Shimizu, sp. nov.), seven are new records from Japan (E. jilinensis Tang, 1990, E. laqueatus (Enderlein, 1921), E. multidens Chiu, 1954, stat. rev., E. puncticulatus Tang, 1990, E. stenophleps Cushman, 1937, E. vestigator (Smith, 1858), and E. zeugos Chiu, 1954, stat. rev.), 32 had already been recorded in Japan; three (E. biharensis Townes, Townes & Gupta, 1961, E. flavicaput (Morley, 1912), and E. merdarius (Gravenhorst, 1829)) have been erroneously recorded from Japan based on misidentifications, and four names that were previously on the Japanese list are deleted through synonymy. The following taxonomic changes are proposed: E. vacuus Gauld & Mitchell, 1981, syn. nov. (= E. formosensis (Uchida, 1928)); E. multidensstat. rev.; E. striatus Cameron, 1899, syn. nov. = E. lineolatus (Roman, 1913), syn. nov. = E. uniformis Chiu, 1954, syn. nov. = E. flatus Chiu, 1954, syn. nov. = E. gussakovskii Viktorov, 1957, syn. nov. = E. striolatus Townes, Townes & Gupta, 1961, syn. nov. = E. unicornis Rao & Nikam, 1969, syn. nov. = E. unicornis Rao & Nikam, 1970, syn. nov. (= E. pungens (Smith, 1874)); E. iracundus Chiu, 1954, syn. nov. (= E. sakaguchii (Matsumura & Uchida, 1926)); E. sigmatoides Chiu, 1954, syn. nov. (= E. shikokuensis (Uchida, 1928)); E. yamanakai (Uchida, 1930), syn. nov. (= E. shinkanus (Uchida, 1928)); E. ranunculus Chiu, 1954, syn. nov. (= E. yezoensis (Uchida, 1928)); and E. zeugosstat. rev. = E. henrytownesi Chao & Tang, 1991, syn. nov. In addition, the following new regional and country records are also provided: E. flavocephalus (Kirby, 1900), E. puncticulatus, and E. vestigator from the Eastern Palaearctic region, E. laqueatus from the Eastern Palaearctic and Oceanic regions, and E. maruyamanus (Uchida, 1928) from the Oriental region; E. abdominalis (Szépligeti, 1906) from Nepal, E. flavocephalus from Laos, E. formosensis from Laos and Malaysia, E. insinuator (Smith, 1860) from Taiwan, E. maruyamanus from India and Philippines, E. nigronotatus Cameron, 1903, E. riukiuensis (Matsumura & Uchida, 1926), and E. sakaguchii from Indonesia, E. pungens from 14 countries (Australia, Bhutan, Brunei, Indonesia, Laos, Malaysia, Nepal, New Caledonia, Papua New Guinea, Philippines, Solomon Islands, Sri Lanka, Tajikistan, and Taiwan), and E. yezoensis from South Korea. An identification key to all Japanese species of Enicospilus is proposed. Although 47 species are recognised in the present study, approximately 55 species could potentially be found in Japan based on ACE and Chao 1 estimators. The latitudinal diversity gradient of Enicospilus species richness is also tested in the Japanese archipelago based on the constructed robust taxonomic framework and extensive samples. Enicospilus species richness significantly increases towards the south, contrary to the 'anomalous' pattern of some other ichneumonid subfamilies.

Matched MeSH terms: Lepidoptera
Vairimorpha imperfecta n.sp., a microsporidian exhibiting an abortive octosporous sporogony in Plutella xylostella L. (Lepidoptera: Yponomeutidae)

Canning EU, Curry A, Cheney S, Lafranchi-Tristem NJ, Haque MA

Parasitology, 1999 Sep;119 ( Pt 3):273-86.
PMID: 10503253

The microsporidian genus Nosema is characterized by development in direct control with host cell cytoplasm, diplokaryotic nuclei throughout development and disporous sporogony. The genus Vairimorpha exhibits the same features plus an octoporous sporogony producing uninucleate spores in a sporophorous vesicle. A microsporidium from diamondback moth, Plutella xylostella, falls between Nosema and Vairimorpha in that it initiates but fails to complete the octosporous sequence in this host. The name Vairimorpha imperfecta n.sp. is proposed. Merogony is mainly by formation of buds from multinucleate meronts, the buds remaining attached in chains. Diplokaryotic spores measure 4.3 x 2.0 microns (fresh) and have 15.5 coils of the polar tube in 1 rank. The octosporous sporogony is aborted owing to irregular formation of nuclear spindles, incomplete cytoplasmic fission and bizarre deposition of electron-dense episporontal secretions. Phylogenetic analyses of the sequences of the small subunit rRNA genes of V. imperfecta and of several Nosema and Vairimorpha spp. place V. imperfecta in a clade with Nosema spp. from Lepidoptera rather than in the clade containing the more typical species of Vairimorpha. It is suggested that the ancestors of the Vairimorpha/Nosema complex of species exhibited both disporous and octosporous sporogonies, as does the type species of Vairimorpha, Vairimorpha necatrix. It would follow that true Nosema spp. have lost the ability to express an octosporous sequence and that V. imperfecta is in the process of losing it. It is proposed that the genera Nosema and Vairimorpha be placed in the same family Nosematidae Labbé 1899, rather than in separate families and orders as at present.

Matched MeSH terms: Lepidoptera/parasitology*
Molecular characterization and enzyme inhibition studies of NADP+- farnesol dehydrogenase from diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae)

Zifruddin AN, Mohamad-Khalid KA, Suhaimi SA, Mohamed-Hussein ZA, Hassan M

Biosci Biotechnol Biochem, 2021 Jun 24;85(7):1628-1638.
PMID: 33890631 DOI: 10.1093/bbb/zbab072

Juvenile hormone III (JH III) plays an important role in insect reproduction, development, and behavior. The second branch of JH III production includes oxidation of farnesol to farnesal by farnesol dehydrogenase. This study reported the identification and characterization of Plutella xylostella farnesol dehydrogenase (PxFoLDH). Our results showed that PxFoLDH belongs to the short-chain dehydrogenase/reductase superfamily, consisting of a single domain with a structurally conserved Rossman fold, an NAD(P) (H)-binding region and a structurally diverse C-terminal region. The purified enzyme displayed maximum activity at 55$\ $°C with pH 9.5 and was stable in the temperature below 70$\ ^\circ $C. PxFoLDH was determined to be a monomer with a relative molecular weight of 27 kDa and highly specific for trans, trans-farnesol, and NADP+. Among analog inhibitors tested, farnesyl acetate was the most effective inhibitor with the lowest Ki value of 0.02 µm. Our findings showed this purified enzyme may represent as NADP+-farnesol dehydrogenase.

Matched MeSH terms: Lepidoptera/enzymology*
Phage displayed Bacillus thuringiensis Cry1Ba4 toxin is toxic to Plutella xylostella

Nathan S, Aziz DH, Mahadi NM

Curr Microbiol, 2006 Nov;53(5):412-5.
PMID: 17036210

We constructed recombinant phage particles displaying the Bacillus thuringiensis Cry1Ba4 active toxin using the pfUSE5 and pComb3X phagemid vectors. The recombinant phage particles were screened and evaluated for displayed biologically active Cry1Ba4 toxin against the target insect larvae. Concurrent expression of Cry1Ba4 protoxin was carried out using the pETBlue -2 plasmid expression vector in Escherichia coli Tuner (DE3)pLacI and the protoxin was successfully expressed at a size of 129 kDa. In the bioassay, 3.30 mg crude extract of Cry1Ba4 protoxin, 9.35 x 10(9) TU and 7.70 x 10(9) TU of induced recombinant phage particles carrying Cry1Ba4 active toxin displayed on pComb3X and pFUSE5, respectively, demonstrated mortality of greater than 85% against Plutella xylostella (third-instar) within 48 hours. Thus, we have successfully displayed the Cry1Ba4 activated toxin on the surface of a phage and demonstrated toxicity towards larvae.

Matched MeSH terms: Lepidoptera