Optimizing the releasing strategy used for the biological control of the sugarcane borer Diatraea saccharalis by Trichogramma galloi with computer modeling and simulation.

One of the challenges in augmentative biological control programs is the definition of releasing strategy for natural enemies, especially when macro-organisms are involved. Important information about the density of insects to be released and frequency of releases usually requires a great number of experiments, which implies time and space that are not always readily available. In order to provide science-based responses for these questions, computational models offer an in silico option to simulate different biocontrol agent releasing scenarios. This allows decision-makers to focus their efforts to more feasible options. The major insect pest in sugarcane crops is the sugarcane borer Diatraea saccharalis, which can be managed using the egg parasitoid Trichogramma galloi. The current strategy consists in releasing 50,000 insects per hectare for each release, in three weekly releases. Here, we present a simulation model to check whether this releasing strategy is optimal against the sugarcane borer. A sensitive analysis revealed that the population of the pest is more affected by the number of releases rather than by the density of parasitoids released. Only the number of releases demonstrated an ability to drive the population curve of the pest towards a negative growth. For example, releasing a total of 600,000 insects per hectare in three releases led to a lower pest control efficacy that releasing only 250,000 insects per hectare in five releases. A higher number of releases covers a wider range of time, increasing the likelihood of releasing parasitoids at the correct time given that the egg stage is short. Based on these results, it is suggested that, if modifications to the releasing strategy are desired, increasing the number of releases from 3 to 5 at weekly intervals is most likely preferable.

Development and characterization of microsatellite markers for population genetics of the cocoa pod borer Conopomorpha cramerella (Snellen) (Lepidoptera: Gracillaridae).

The cocoa pod borer (CPB) Conopomorpha cramerella (Snellen) (Lepidoptera: Gracillaridae) is one of the major constraints for cocoa production in South East Asia. In addition to cultural and chemical control methods, autocidal control tactics such as the Sterile Insect Technique (SIT) could be an efficient addition to the currently control strategy, however SIT implementation will depend on the population genetics of the targeted pest. The aim of the present work was to search for suitable microsatellite loci in the genome of CPB that is partially sequenced. Twelve microsatellites were initially selected and used to analyze moths collected from Indonesia, Malaysia, and the Philippines. A quality control verification process was carried out and seven microsatellites found to be suitable and efficient to distinguish differences between CPB populations from different locations. The selected microsatellites were also tested against a closely related species, i.e. the lychee fruit borer Conopomorpha sinensis (LFB) from Vietnam and eight loci were found to be suitable. The availability of these novel microsatellite loci will provide useful tools for the analysis of the population genetics and gene flow of these pests, to select suitable CPB strains to implement the SIT.

The good, the bad and the boa: An unexpected new species of a true boa revealed by morphological and molecular evidence.

Snakes of the genus Boa are outstanding elements of the New World biota with a broad sociological influence on pop culture. Historically, several taxa have been recognized in the past 300 years, being mostly described in the early days of binomial nomenclature. As a rule, these taxa were recognized based on a suite of phenotypic characters mainly those from the external morphology. However, there is a huge disagreement with respect to the current taxonomy and available molecular phylogenies. In order to reconcile both lines of evidence, we investigate the phylogenetic reconstruction (using mitochondrial and nuclear genes) of the genus in parallel to the detailed study of some phenotypic systems from a geographically representative sample of the cis-Andean mainland Boa constrictor. We used cyt-b only (744bp) from 73 samples, and cyt-b, ND4, NTF3, and ODC partial sequences (in a total of 2305 bp) from 35 samples, comprising nine currently recognized taxa (species or subspecies), to infer phylogenetic relationships of boas. Topologies recovered along all the analyses and genetic distances obtained allied to a unique combination of morphological traits (colouration, pholidosis, meristic, morphometric, and male genitalia features) allowed us to recognize B. constrictor lato sensu, B. nebulosa, B. occidentalis, B. orophias and a distinct lineage from the eastern coast of Brazil, which we describe here as a new species, diagnosing it from the previously recognized taxa. Finally, we discuss the minimally necessary changes in the taxonomy of Boa constrictor complex; the value of some usually disregarded phenotypic character system; and we highlight the urgency of continuing environmental policy to preserve one of the most impacted Brazilian hotspots, the Atlantic Forest, which represents an ecoregion full of endemism.

Spatio-temporal classification of land use and land cover and its changes in Kerala using remote sensing and machine learning approach.

Land use and land cover (LULC) analysis gives important information on how the region has evolved over time. Kerala, a land with an extensive and dynamic history of land-use changes, has, until now, lacked comprehensive investigations into this history. So the current study focuses on Kerala, one of the ecologically diverse states in India with complex topography, through Landsat images taken from 1990 to 2020 using two different machine learning classifications, random forest (RF) and classification and regression trees (CART) on Google Earth Engine (GEE) platform. RF and CART are versatile machine learning algorithms frequently employed for classification and regression, offering effective tools for predictive modelling across diverse domains due to their flexibility and data-handling capabilities. Normalised Difference Vegetation Index (NDVI), Normalised Differences Built-up Index (NDBI), Modified Normalised Difference Water Index (MNDWI), and Bare soil index (BSI) are integral indices utilised to enhance the precision of land use and land cover classification in satellite imagery, playing a crucial role by providing valuable insights into specific landscape attributes that may be challenging to identify using individual spectral bands alone. The results showed that the performance of RF is better than that of CART in all the years. Thus, RF algorithm outputs are used to infer the change in the LULC for three decades. The changes in the NDVI values point out the loss of vegetation for the urban area expansion during the study period. The increasing value of NDBI and BSI in the state indicates growth in high-density built-up areas and barren land. The slight reduction in the value of MNDWI indicates the shrinking water bodies in the state. The results of LULC showed the urban expansion (158.2%) and loss of agricultural area (15.52%) in the region during the study period. It was noted the area of the barren class, as well as the water class, decreased steadily from 1990 to 2020. The results of the current study will provide insight into the land-use planners, government, and non-governmental organizations (NGOs) for the necessary sustainable land-use practices.

Biochemistry and transcriptomic analyses of Phthorimaea absoluta (Lepidoptera: Gelechiidae) response to insecticides.

Phthorimaea absoluta is an invasive solanaceous plant pest with highly devastating effects on tomato plant. Heavy reliance on insecticide use to tackle the pest has been linked to insecticide resistance selection in P. absoluta populations. To underline insights on P. absoluta insecticide resistance mechanisms to diamides and avermectins, we evaluated the transcriptomic profile of parental (field-collected) and F8 (lab-reared) populations. Furthermore, to screen for the presence of organophosphate and pyrethroid resistance, we assessed the gene expression levels of acetylcholinesterase (ace1) and para-type voltage-gated sodium channel (VGSG) genes in the F1 to F8 lab-reared progeny of diamide and avermectin exposed P. absoluta field-collected populations. The VGSG gene showed up-regulation in 12.5% and down-regulation in 87.5% of the screened populations, while ace1 gene showed up-regulation in 37.5% and down-regulation in 62.5% of the screened populations. Gene ontology of the differentially expressed genes from both parental and eighth generations of diamide-sprayed P. absoluta populations revealed three genes involved in the metabolic detoxification of diamides in P. absoluta. Therefore, our study showed that the detoxification enzymes found could be responsible for P. absoluta diamide-based resistance, while behavioural resistance, which is stimulus-dependent, could be attributed to P. absoluta avermectin resistance.

The BTB-ZF gene Bm-mamo regulates pigmentation in silkworm caterpillars.

The color pattern of insects is one of the most diverse adaptive evolutionary phenotypes. However, the molecular regulation of this color pattern is not fully understood. In this study, we found that the transcription factor Bm-mamo is responsible for black dilute (bd) allele mutations in the silkworm. Bm-mamo belongs to the BTB zinc finger family and is orthologous to mamo in Drosophila melanogaster. This gene has a conserved function in gamete production in Drosophila and silkworms and has evolved a pleiotropic function in the regulation of color patterns in caterpillars. Using RNAi and clustered regularly interspaced short palindromic repeats (CRISPR) technology, we showed that Bm-mamo is a repressor of dark melanin patterns in the larval epidermis. Using in vitro binding assays and gene expression profiling in wild-type and mutant larvae, we also showed that Bm-mamo likely regulates the expression of related pigment synthesis and cuticular protein genes in a coordinated manner to mediate its role in color pattern formation. This mechanism is consistent with the dual role of this transcription factor in regulating both the structure and shape of the cuticle and the pigments that are embedded within it. This study provides new insight into the regulation of color patterns as well as into the construction of more complex epidermal features in some insects.

Identifying Areas of Invasion Risk and Changes in the Ecological Niche Occupied by the Coffee Leaf Miner Leucoptera coffeella (Lepidoptera: Lyonetiidae).

Insects of economic importance such as Leucoptera coffeella can cause high defoliation in plants and reduce crop yields. We aimed to identify changes in the ecological niche and potential zones of the invasion. Occurrence records were obtained from databases and bibliography. WorldClim V2.0 bioclimatic layers were used. For the modeling of the potential distribution, the kuenm R package was used by executing the Maxent algorithm. The potential distribution models suggested greatest environmental suitability extends from Europe, South Asia, and Central and South Africa, showing the "tropical and subtropical moist broadleaf forests" as the ecoregion that presents the greatest probability of the presence of L. coffeella. The potential distribution model projected in the invaded area agrees with the known distribution in the region (America), although the results show that it is occupying environmental spaces not present in the area of origin. This species presented a large proportion of the invaded niche that overlaps the native niche and is colonizing new environmental conditions in the invaded area relative to its native distribution (Africa). This information could be used in the planning of coffee crops on the American continent.

Effects of climate change on Lepidoptera pollen loads and their pollination services in space and time.

Shifts in flowering time among plant communities as a result of climate change, including extreme weather events, are a growing concern. These plant phenological changes may affect the quantity and quality of food sources for specialized insect pollinators. Plant-pollinator interactions are threatened by habitat alterations and biodiversity loss, and changes in these interactions may lead to declines in flower visitors and pollination services. Most prior research has focused on short-term plant-pollinator interactions, which do not accurately capture changes in pollination services. Here, we characterized long-term plant-pollinator interactions and identified potential risks to specialized butterfly species due to habitat loss, fragmented landscapes, and changes in plant assemblages. We used 21 years of historical data from museum specimens to track the potential effects of direct and indirect changes in precipitation, temperature, monsoons, and wildfires on plant-pollinator mutualism in the Great Basin and Sierra Nevada. We found decreased pollen richness associated with butterflies within sites, as well as an increase in pollen grain abundance of drought-tolerant plants, particularly in the past 10 years. Moreover, increased global temperatures and the intensity and frequency of precipitation and wildfires were negatively correlated with pollen diversity. Our findings have important implications for understanding plant-pollinator interactions and the pollination services affected by global warming.

The complete mitochondrial genome of Leucoptera coffeella (Lepidoptera: Lyonetiidae) and phylogenetic relationships within the Yponomeutoidea superfamily.

The coffee leaf miner (Leucoptera coffeella) is one of the major pests of coffee crops in the neotropical regions, and causes major economic losses. Few molecular data are available to identify this pest and advances in the knowledge of the genome of L. coffeella will contribute to improving pest identification and also clarify taxonomy of this microlepidoptera. L. coffeella DNA was extracted and sequenced using PacBio HiFi technology. Here we report the complete L. coffeella circular mitochondrial genome (16,407 bp) assembled using Aladin software. We found a total of 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), 2 ribosomal RNA genes (rRNAs) and an A + T rich-region and a D-loop. The L. coffeella mitochondrial gene organization is highly conserved with similarities to lepidopteran mitochondrial gene rearrangements (trnM-trnI-trnQ). We concatenated the 13 PCG to construct a phylogenetic tree and inferred the relationship between L. coffeella and other lepidopteran species. L. coffeella is found in the Lyonetiidae clade together with L. malifoliella and Lyonetia clerkella, both leaf miners. Interestingly, this clade is assigned in the Yponomeutoidea superfamily together with Gracillariidae, and both superfamilies displayed species with leaf-mining feeding habits.

The complete mitochondrial genome of Chibiraga houshuaii (Lepidoptera, Limacodidae) and its phylogenetic implications.

Chibiraga is a mall East Asian genus in the family Limacodidae (slug-moths). The latter includes many agricultural pests. Mitochondrial genome analysis is an important tool for studying insect molecular identification and phylogenetics. However, there are very few mitogenome sequences available for Limacodidae species, and none for the genus Chibiraga at all. To explore the mitogenome features of Chibiraga and verify its phylogenetic position, the complete mitogenome of Chibiraga houshuaii was sequenced and annotated. The complete 15,487 bp genome encoded 37 mitochondrial genes, including 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and a control region (CR). Most of the PCGs had typical ATN start codons and terminated with TAA or a single T residue. UUA (Leu2), AUU (Ile), UUU (Phe), AUA (Met) and AAU (Asn) were the five most frequently used codons. All tRNAs were folded into cloverleaf secondary structure, except for trnS1, which lacked the DHU arm. Phylogenetic analyses within the superfamily Zygaenoidea were performed based on multiple datasets from mitochondrial genes. The results showed that the families Phaudidae, Limacodidae and Zygaenidae were respectively recovered as monophyly; C. houshuaii was clustered in a clade with nettle type larvae in Limacodidae.

GC-MS-based Metabolomics Unravels Metabolites across Larval Development and Diapause of a Specialist Insect.

Plant-insect interactions are a driving force into ecosystem evolution and community dynamics. Many insect herbivores enter diapause, a developmental arrest stage in anticipation of adverse conditions, to survive and thrive through seasonal changes. Herein, we investigated the roles of medium- to non-polar metabolites during larval development and diapause in a specialist insect herbivore, Chlosyne lacinia, reared on Aldama robusta leaves. Varying metabolites were determined using gas chromatography-mass spectrometry (GC-MS)-based metabolomics. Sesquiterpenes and steroids were the main metabolites putatively identified in A. robusta leaves, whereas C. lacinia caterpillars were characterized by triterpenes, steroids, fatty acids, and long-chain alkanes. We found out that C. lacinia caterpillars biosynthesized most of the identified steroids and fatty acids from plant-derived ingested metabolites, as well as all triterpenes and long-chain alkanes. Steroids, fatty acids, and long-chain alkanes were detected across all C. lacinia instars and in diapausing caterpillars. Sesquiterpenes and triterpenes were also detected across larval development, yet they were not detected in diapausing caterpillars, which suggested that these metabolites were converted to other molecules prior to the diapause stage. Our findings shed light on the chemical content variation across C. lacinia development and diapause, providing insights into the roles of metabolites in plant-insect interactions.

A new orchid species expands Darwin's predicted pollination guild in Madagascar.

The world-renowned pollination system of the long-spurred orchid Angraecum sesquipedale Thouars and the long-tongued hawkmoth Xanthopan praedicta (Rothschild & Jordan, 1903), from Madagascar, is the best-known example of the predictive power of evolutionary ecology1,2, yet its actual degree of specialisation remains poorly described due to the incompleteness of the pollination record of X. praedicta. Here, we describe another species from Madagascar, an angraecoid orchid distantly related to the genus Angraecum Bory, that has evolved these extreme adaptations to a single pollinator after a pollinator shift. It bears the longest spur of any flowering plant, relative to flower diameter, reaching 33 cm. The discovery of a species with such an exceptionally long spur is a rare event, the most recent dating to 19653. This novelty is described here as Solenangis impraedicta (Figure 1A-F) and discussed in a phylogenetic framework. Its conservation status is assessed as Endangered.

A DNA barcode library of Austrian geometridae (Lepidoptera) reveals high potential for DNA-based species identification.

Situated in the Eastern section of the European Alps, Austria encompasses a great diversity of different habitat types, ranging from alpine to lowland Pannonian ecosystems, and a correspondingly high level of species diversity, some of which has been addressed in various DNA barcoding projects. Here, we report a DNA barcode library of all the 476 species of Geometridae (Lepidoptera) that have been recorded in Austria. As far as possible, species were sampled from different Austrian regions in order to capture intraspecific genetic variation. In total, 2500 DNA barcode sequences, representing 438 species, were generated in this study. For complete coverage of Austrian geometrid species in the subsequent analyses, the dataset was supplemented with DNA barcodes from specimens of non-Austrian origin. Species delimitations by ASAP, BIN and bPTP methods yielded 465, 510 and 948 molecular operational taxonomic units, respectively. Congruency of BIN and ASAP partitions with morphospecies assignments was reasonably high (85% of morphospecies in unique partitions), whereas bPTP appeared to overestimate the number of taxonomic units. The study furthermore identified taxonomically relevant cases of morphospecies splitting and sharing in the molecular partitions. We conclude that DNA barcoding and sequence analysis revealed a high potential for accurate DNA-based identification of the Austrian Geometridae species. Additionally, the study provides an updated checklist of the geometrid moths of Austria.

Review of the spider wasp genus Episyron Schidte, 1837 (Hymenoptera: Pompilidae) from India, with description of three new species.

The spider wasp genus Episyron Schidte, 1837 is reviewed, with five species recorded from various parts of India along with three new species described and illustrated from Kerala: E. keralaensis Anju, Girish Kumar & Thejass, sp. nov., E. nigrocalcarius Anju, Girish Kumar & Thejass, sp. nov., and E. rufotibius Anju, Girish Kumar & Thejass, sp. nov. Diagnosis of species occurring in India, comparisons of new species with congeners and a key to the Indian species also are provided.

Review of the genus Antoniejanse Kemal & Koak, 2005 (Lepidoptera, Gelechiidae, Litini), with description of four new species.

The southern African genus Antoniejanse Kemal & Koak, 2005 is revised. Seven species are recognized here as valid, of which four species are described as new for science from South Africa (A. varii sp. nov. and A. tenebrosa sp. nov.) and Namibia (A. minima sp. nov. and A. mediopunctata sp. nov.). One new combination is proposed: Antoniejanse sagittata (Bidzilya, 2019) comb. nov. (ex Paratelphusa). All species of Antoniejanse are re-diagnosed. Wing patterns, as well as male and female genitalia are depicted. An identification key to all species of the genus is provided and the checklist of the genus is updated.

Two new species of the genus Nosphistica Meyrick (Lepidoptera, Lecithoceridae, Lecithocerinae) from China, with a checklist of the world.

Two new species of the genus Nosphistica Meyrick, 1911 from China are described as new: N. eucalla sp. nov. and N. longiclavata sp. nov. Images of adults and genitalia for the newly described species and a checklist of the genus are given.

Two new species of Cameraria Chapman (Lepidoptera: Gracillariidae) associated with Salix or Cornus in Japan.

Two new species of the genus Cameraria Chapman, 1902, Cameraria riparia sp. nov. and Cameraria corni sp. nov., are described from Japan. Larval setal maps, illustrations and/or photographs of wing patterns, wing venations and genitalia are provided for both species, and the systematic positions of the two species are discussed based on their morphologies. The larvae of C. riparia feed on Salix spp. and make blotch mines, whereas C. corni larvae feed on Cornus kousa subsp. kousa and make linear blotch mines. C. riparia was collected on Hokkaido and Honshu Islands, and considerable differences in the male genital characters were detected between the two populations. However, a molecular phylogenetic analysis based on the COI barcode region indicated that there was no clear genetic differentiation between the two populations (maximum divergence, 1.41%). C. corni was collected on Honshu and Kyushu Islands, and this is the first record of the trophic association of Lithocolletinae with Cornaceae.

Description of two new species of the genus Repetekiodes Amsel, 1961 (Lepidoptera: Pyralidae: Phycitinae) from Central Asia with faunal notes on the genus.

Two new species, Repetekiodes serratalis sp. nov. and Repetekiodes turanella sp. nov. are described from Central Asia. The described species are well distinguished from their congeners by the genitalia and forewing pattern. A brief overview of the genus Repetekiodes Amsel, 1961 is given and original data on the distribution of Repetekiodes species are provided. DNA barcode data are presented for several species of the genus.

Fairy moths of the genus Nemophora Hoffmannsegg, 1798 (Lepidoptera: Adelidae) from Myanmar, Thailand, Laos, Cambodia and Vietnam.

This study provides a comprehensive account of 40 species (52 valid names and one preoccupied name) of the genus Nemophora Hoffmannsegg, 1798 described or recorded from Myanmar, Thailand, Laos, Cambodia and Vietnam. A key to the species based on external characters and on male genitalia is provided; 14 new species are described: N. auricapitella Kozlov, sp. nov., N. chalcoptera Kozlov, sp. nov., N. karsholti Kozlov, sp. nov., N. kuznetzovi Kozlov, sp. nov., N. meyi Kozlov, sp. nov., N. nielseni Kozlov, sp. nov., N. nieukerkeni Kozlov, sp. nov., N. nigripunctella Kozlov, sp. nov., N. punctifasciella Kozlov, sp. nov., N. rubicunda Kozlov, sp. nov., N. szabokyi Kozlov, sp. nov., N. thailandensis Kozlov, sp. nov., N. vietnamensis Kozlov, sp. nov. and N. yeni Kozlov, sp. nov. Lectotypes are designated for four species, including one junior synonym: Nemotois diplophragma Meyrick, 1938, Nemotois sinicella Walker, 1863, Nemotois cleodoxa Meyrick, 1922 and Adela satrapodes Meyrick, 1894. Eight new synonyms for four species are proposed: Nemophora badagongshana Liao, Hirowatari & Huang in Liao, Hirowatari, Yagi, Wang, Wang & Huang, 2023, syn. nov. of Nemophora aurifera (Butler, 1881); Nemotois chrysocharis Caradja, 1938, syn. nov. of Nemophora sinicella (Walker, 1863); Nemotois diplophragma Meyrick, 1938, syn. nov. of Nemophora melichlorias (Meyrick, 1907); Nemophora basalistriata Liao, Hirowatari & Huang in Liao, Hirowatari, Yagi, Wang, Wang & Huang, 2023, syn. nov. of Nemophora melichlorias (Meyrick, 1907); Nemotois limenites Meyrick, 1914, syn. nov. of Nemophora aurifera (Butler, 1881); Nemophora quadrata Liao, Hirowatari & Huang in Liao, Hirowatari, Yagi, Wang, Wang & Huang, 2023, syn. nov. of Nemophora aurifera (Butler, 1881); Nemotois rubrofascia formosicola Matsumura, 1931, syn. nov. of Nemophora sakaii (Matsumura, 1931); Nemotois servata Meyrick in Caradja, 1925, syn. nov. of Nemophora aurifera (Butler, 1881). Four new combinationsare introduced: Nemophora aglaospila (Meyrick, 1928), comb. nov.; Nemophora cleodoxa (Meyrick, 1922), comb. nov.; Nemophora melichlorias (Meyrick, 1907), comb. nov.; and Nemophora sinicella (Walker, 1863), comb. nov. Of 40 species considered, 16 were recorded in Myanmar, 21 in Thailand, 4 in Laos, 4 in Cambodia and 13 in Vietnam.