The stingless bee Geniotrigona thoracica is an ecologically and economically important pollinator in Southeast Asia, yet comprehensive genomic resources for this species remain limited. In this study, we sequenced, assembled, and annotated the complete mitochondrial genome (mitogenome) of G. thoracica to investigate its genomic architecture and phylogenetic position. The circular mitogenome is 16,061 bp in length and comprises the typical set of 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, and two ribosomal RNA genes. The genome exhibits a strong A + T bias, consistent with other hymenopteran mitogenomes, and codon usage patterns reflect this nucleotide composition. Most tRNAs display the canonical cloverleaf secondary structure, although minor structural variations were observed. Comparative analyses revealed several gene rearrangements, including transposition and inversion events, suggesting lineage-specific rearrangements, including transposition of the cox1-trnL-cox2-trnD-atp8-atp6-cox3 block and transposition with inversion of the trnF-nad5-nad4-nad4l-trnP block, relative to the ancestral hymenopteran gene order. Phylogenomic analyses based on concatenated mitochondrial genes strongly supported the monophyly of Meliponini and placed G. thoracica within a well-supported Indo-Malayan clade, closely related to Tetragonula, Heterotrigona, and Lepidotrigona. Furthermore, stingless bees were recovered as more closely related to bumblebees than to honeybees, consistent with previous studies. Overall, this study provides a complete, annotated mitogenomic resource for G. thoracica and contributes to a better understanding of mitochondrial genome evolution, phylogenetic relationships, and biogeographic patterns in stingless bees.

The coffee berry borer (Hypothenemus hampei) is one of the most destructive pests of coffee worldwide, causing severe economic losses. The entomopathogenic fungus Beauveria bassiana has been widely used as a biological control agent; however, detailed histopathological responses of H. hampei to fungal infection remain poorly understood. This study investigated the morpho-histopathological alterations and apoptosis in adult H. hampei following exposure to B. bassiana. Beetles were immersed in a spore suspension (1×10⁹ conidia/mL) and sampled at multiple time points up to 144 h post-exposure. Histological analyses were performed using hematoxylin-eosin, PAS, Masson’s trichrome, and Grocott’s methenamine silver staining. Apoptotic cells were detected using TUNEL assay, and tissue damage was evaluated using the histological alteration index (HAI). Fungal hyphae were detected in the integument and adipose tissue as early as 12 h post-exposure. From 24 to 144 h, progressive fungal proliferation, cuticle thinning, muscle degeneration, adipose tissue degeneration, and gut epithelial erosion were observed. Quantitative analysis revealed a significant increase in fungal area and a marked reduction in cuticle thickness (p < 0.001). Apoptotic cell density increased in a time-dependent manner, with strong immunoreactivity observed after 60 h. These findings demonstrate that B. bassiana rapidly penetrates and disrupts host tissues, leading to extensive structural damage and apoptosis. Understanding the infection dynamics of B. bassiana provides important insights for optimizing biological control strategies and enhancing sustainable pest management in coffee agroecosystems. HIGHLIGHTS First detailed morpho-histopathological analysis ofBeauveria bassiana infection in Hypothenemus hampei. Rapid fungal penetration of the cuticle and adipose tissue were observed within 12 h post-exposure. Significant reduction in cuticle thickness and expansion of fungal area from 48 - 144 h. Time-dependent increase in histological alteration index (HAI) and apoptotic cell density. Findings clarify infection dynamics supporting the biological control development. GRAPHICAL ABSTRACT

The marine water-strider Halobates hayanus is prevalent in tropical and subtropical marine habitats. Although the ecology and the morphology of this insect are widely accepted, some histological data remain limited. In this study, we determined new data on the organ histological development for specific performance of H. hayanus, locally collected with respect to its total length (TL) from Libong Island, Thailand. Our observation revealed that the TL of this insect could be divided into three groups: (1) Small, 0.1-0.2 cm; (2) Medium, 0.3-0.4 cm; and (3) Large, 0.5-0.6 cm. Histologically, subsequent microscopic development of eyes, Malphigian tubules, midgut, and gonads of H. hayanus were found in Group (3), 0.5-0.6 cm in TL. Surprisingly, the distinctive structure of pigment cells, with strong pigment granules and crystalline cones, was also found in the 0.5-0.6 cm group. Clear mature testes with the presence of sperms were evident in the 0.1-0.2 cm group, and the sampled females from the 0.5-0.6 cm group had mature ovaries with the presence of the vitellogenic stage. The results obtained from this study expanded the histological data of H. hayanus from natural environments, contributing to further determination of the insect's development stages and physiology.

The stingless bee Geniotrigona thoracica is a key managed pollinator in Southeast Asia, valued for its honey, propolis, and colony trade. In Thailand, frequent human-mediated movement of colonies raises concerns about its effects on genetic diversity and population structure. We analysed variation in mitochondrial (COI and 16S rRNA) and nuclear (five microsatellite loci) markers from 70 colonies sampled across 17 meliponaries in seven southern provinces. Microsatellite data revealed high genetic diversity and low nuclear differentiation (K = 1; Fst = 0.0024–0.1219; all P > 0.05), with extensive gene flow (Nm = 3.60–207.83) among provinces. In contrast, mitochondrial markers indicated moderate-to-high differentiation (Fst = 0.619), consistent with mito-nuclear discordance arising from sex-biased. Managed colonies exhibited elevated heterozygosity and allelic richness, likely reflecting admixture from colony exchange, while unique haplotypes in certain provinces suggest introductions from external sources. Significant inbreeding was detected only in Yala, possibly linked to habitat loss and reduced effective population size. Our findings indicate that current meliponicultural practices maintain high genetic diversity in G. thoracica despite mitochondrial structuring, but increasing colony movement between genetically distinct populations may risk erosion of local adaptations, underscoring the need for genetic screening prior to translocation.

The use of entomopathogenic fungi (EPF), such as Cordyceps javanica, to reduce insect pest populations is gaining traction since it is an environmentally safe approach that can control many pests at different life stages. Here, we focus on the histopathology of the coffee berry borer, Hypothenemus hampei, infected by C. javanica. Morphological observation revealed that C. javanica conidia germinated within 12 h following inoculation according to light microscopic and ultrastructural levels. The fungus thoroughly penetrated the fat body and muscular tissue between 84 and 120 h post-inoculation. Transmission electron microscopy (TEM) confirmed the hyphal invasion of the cuticle at 12 h post-inoculation, with progressive tissue disruption and organelle degeneration, especially mitochondria and rough endoplasmic reticulum in adipocytes. All organelles were completely degenerated at 96 h post-inoculation. There was evidence of a connection between C. javanica activity and the coffee berry borer that might cause histopathological changes in the host defense against the pathogen, pointing to increased mortality and potential control of coffee berry borer in natural populations. Additionally, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) confirmed that apoptotic cells were slightly increased in the adipose tissue and integument of the coffee berry borer. The ability of C. javanica to fatally infect the coffee berry borer suggests that it could be deployed as a biological control agent in the field.

Metarhizium guizhouense (Clavicipitaceae) is an insect pathogen employed as a mycoinsecticide against many insect pests worldwide. In this study, for the first time in a laboratory setting, the pathogenicity of the M. guizhouense isolate PSUM04 was evaluated against the coffee berry borer, Hypothenemus hampei (Coleoptera: Curculionidae: Scolytinae). Following the exposure (1 × 10 9 spores/mL) of H. hampei to M. guizhouense PSUM04, morpho-histopathological changes and numbers of apoptotic cells via the TUNEL (terminal deoxynucleotidyl transferase (TdT), dUTP (deoxyuridine triphosphate) nick-end labelling) assay in H. hampei were evaluated at 12-hour intervals until 144 hours after exposure. The external morphology of H. hampei underwent drastic changes from 24 to 144 hours post-exposure accompanied by the degeneration of integument and adipose tissues. The semi-quantitative analytical score of TUNEL-positive cells showed a slight increase at 12 hours after exposure and a significant increase in TUNEL-positive apoptotic cells at 48 hours after exposure. These results suggest a series of tissue alterations of H. hampei during its process of infection with a strain of M. guizhouense , highlighting the pathogen’s potential as a biological control agent in natural settings.

With increasing demand for stingless bee honey, meliponiculture has gained widespread attention. Heterotrigona itama is one of the most economically important species. However, excessive exploitation for commercial purposes has led to population declines, and the species is now considered vulnerable in Thailand. Despite its ecological and economic significance, genomic and taxonomic information on H. itama remains limited. In this study, we sequenced and characterized the complete mitochondrial genome (mitogenome) of H. itama to explore its genome structure and phylogenetic position. The circular mitogenome is 15,318 bp in length and consists of 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, and 2 ribosomal RNA (rRNA) genes. The genome exhibits a strong A+T bias (75.41%), which affects codon usage and amino acid composition. Isoleucine, methionine, and phenylalanine were the most commonly encoded amino acids. Gene arrangement was highly conserved and closely resembled that of Tetragonula species. Phylogenetic analyses confirmed that H. itama clusters with other stingless bees and is more closely related to bumblebees than to honeybees. Several gene rearrangements suggest a high degree of mitogenomic plasticity. This study provides essential genomic resources for future studies in systematics, phylogenetics, population genetics, and conservation of stingless bees in the Meliponini tribe.

Stingless beekeeping, also known as meliponiculture, has gained increasing popularity in many tropical and subtropical countries for its use in commercial pollination and high-value honey and propolis production. However, this rising interest in stingless beekeeping has led to significant geographical displacements of bee colonies by beekeepers, occasionally surpassing their native ranges. Consequently, this affects local bee populations by disrupting gene flow across unnaturally large geographic scales. For Heterotrigona itama, one of the most common stingless bee species in Southeast Asian countries, including Thailand, there is concern that large-scale artificial propagation by beekeepers utilizing a limited number of bee colonies will lead to inbreeding. This practice leads to increased inbreeding within managed populations and introgression into wild populations. These concerns highlight the need for careful management practices in stingless beekeeping to mitigate potential adverse effects. To assess the genetic structure of H. itama in Thailand, 70 colonies were sampled, and partially sequenced cytochrome c oxidase subunit 1 (COI) gene, large ribosomal subunit rRNA gene (16S rRNA), and 28S large ribosomal subunit rDNA gene (28S rRNA) were analyzed. Our results showed slightly lower nuclear genetic variability, but higher mitochondrial genetic variability, which can be attributed to gene flow, colony transport, and nest division. We suggest that increasing the number of colonies maintained through nest division does not negatively affect genetic variability, as it is maintained by small-scale male dispersal and human-mediated nest transport. However, caution should be exercised when transporting nests from distant localities, considering the high genetic differentiation observed between samples from Narathiwat and those from Krabi and Nakhon Si Thammarat provinces, which might indicate local adaptation.

Polyneopteran insects have relatively large genomes compared to holometabolous insects, which appear constrained by a threshold of 2 pg/1C DNA. This threshold may be due to costly complex development and higher energy demands. Genome sizes (GSs) are particularly bulky in various species of Orthoptera, reaching sizes up to 18.64 pg/1C DNA, however, recent analyses comparing insect GSs neglected the order Embioptera (webspinners). Our access to fresh specimens of 12 species from 6 of the 13 named taxonomic families provided a chance to determine if embiopterans also support bulky genomes. Flow cytometry results revealed that embiopteran GSs ranged from 2.41 to 7.56 pg/1C, similar to other polyneopterans. Based on previous studies reporting correlations with GS, we tested for correlations between GS and body length using independent contrasts. The ancestral state of the root was estimated at 4.57 pg/1C. A positive relationship was detected whereby larger-bodied webspinners displayed larger genomes, with female Antipaluria urichi (Saussure) (Clothodidae) having the largest genome at 7.56 pg/1C. This GS is approximately 3 times larger than the previously reported embiopteran GS of pg = 2.66 for a male Oligotoma saundersii (Westwood) (Oligotomidae). GS showed no consistent patterns of phylogenetic signal for Embioptera. The underlying causes for large genomes are briefly reviewed.

Thai species of the pill millipede genus Hyleoglomeris Verhoeff, 1910 are reviewed, and an annotated catalogue is provided. Six new species from Thailand are described and illustrated based on morphological characteristics and molecular analyses: H. dracosphaera sp. n., H. nigromaculata sp. n., H. suwannakhuhensis sp. n., H. bomba sp. n., H. tongkerdae sp. n., and H. krasoon sp. n. They are morphologically similar, yet all can be separated based on their characteristic colour patterns, telopod structures and geographic distributions. Genetic differences in the dna barcode region confirm their full species statuses, with interspecific coi p-distances ranging from 9.01% to 16.51% dissimilarity. We also propose the following new synonym: H. hongkhraiensis Likhitrakarn, Golovatch & Panha, 2015 = H. cavicola Likhitrakarn, Golovatch & Panha, 2015 syn. n. This is based on the low genetic divergence observed between these two taxa (2.34% coi p-distance). A new distribution map of and a key to all Hyleoglomeris species presently known to occur in Thailand are also provided.

The cartilage and bone structure has provided insightful knowledge about evolution and ecology of fish, which is an important component of biological oceanography. However, the whole-body bone staining is a lengthy and complicated process that typically takes five days to several months, and the improvement of the conventional method has been one of the important issues in this field. Here we report a quick and easy whole-mount bone staining method for small fish, in which a newly designed fixative is applied. Compared to conventional methods, this novel protocol is a straightforward process that could be adopted for small estuarine fish and other small vertebrates.

Identification, evaluation and optimization of a minimum simple sequence repeat marker set for triticale breedingW.C. Botes and D. Bitalo

Stingless bees, prevalent in tropical and subtropical regions, are a tribe of eusocial bees that are crucial pollinators for economic crops and native plants and, produce honey and pollen. However, colony expansion is limited by a shortage of queens for new colonies. Therefore, mass artificial rearing of virgin queens could address this in commercially managed meliponiculture. Furthermore, the in vitro rearing of queen stingless bees can improve meliponiculture management and conservation efforts. Herein, we explored the efficacy of in vitro queen rearing for Heterotrigona itama, assessing the queen's body size, reproductive organ size (ovary and spermatheca), acceptance rate into new, small colonies, and mating frequency. H. itama larvae developed into queens when fed with 120 mu L-150 mu L of larval food, resulting in in vitro queens having body sizes similar to those of naturally produced queens. Microscopic analysis revealed well-developed ovaries and spermathecae in in vitro-reared queens, unlike the smaller ovaries and the absence of spermathecae in the naturally produced workers. Acceptance of in vitro-reared queens was independent of worker age, and mating frequency was low but not significantly different from naturally produced queens. These findings could enhance stingless beekeeping practices and conservation efforts for the native stingless bee species.

The structural evidence of tephritids, notably Bactrocera albistrigata, B. dorsalis, B. umbrosa, Zeugodacus cucurbitae, and Z. tau complex were explored using histology. These tephritids' male reproductive organs comprised two testes, ducts with deferent ducts, seminal vesicles, and tubular exocrine glands. The testicular follicle was investigated as a cyst during four stages of spermatogenesis (spermatogonium, spermatocyte, spermatid, and spermatozoa). Similarly, the female reproductive systems of these fruit flies were morphologically identical, with a pair of ovaries containing seven meroistic ovariole types. There was also a spermatheca, two lateral oviducts, a common oviduct, and a genital chamber identified. Tephritids have digestive systems (foregut, midgut, and hindgut), excretory systems (malpighian tubules), neural systems, integumentary systems (cuticle and epidermis), and adipose tissue. These data are not only significant and publicly available, but they can also be used in future studies such as on histopathology, ecotoxicological assays, and phylogenic characteristics.

Close to industrial activity and a major seaport in the Gulf of Thailand, Sichang Island has frequently suffered from pollution and oil spills. However, the environmental health status of the surrounding waters is relatively unknown. Between December 2017 and January 2018, we reared ninety Asian seabass (Lates calcarifer) in cage nets off the island and sampled blood to investigate nuclear abnormalities in erythrocytes and build leucocyte profiles. The fish were purchased locally and reared in locations where oil spills have been recorded. Environmental parameters were within the range of standard values. Erythrocyte nuclei had segmented, reniform and notched abnormalities; however, they dramatically varied after exposure. At the end of the three-month field experiment, very few micronuclei had been observed. Observed leucocytes were neutrophils, lymphocytes, and monocytes. The percentage of lymphocytes increased considerably, reaching a post-exposure peak at three months. Our data suggest that erythrocytic nuclear abnormalities of Asian seabass could be used as an early warning of toxic pollutants in the marine environment, and as a baseline environmental health indicator for Sichang Island and the surrounding area.

The ambrosia beetle Xylosandrus compactus (Eichhoff, 1875) is a serious pest of economically important plants including Mitragyna speciosa, but its body structure and cellular composition remain uncharacterized. In this study, we described whole-body histological characteristics of female X. compactus. Our observation found two district regions in the nervous system: frontal ganglion and ventral nerve cord. The frontal ganglion contained two layers: outer cortex and inner medullae, and the outer cortex was lined with three major types of cells including neurosecretory cells, neuroglia, and neurons. The integumentary system contained three different layers: epicuticle, exocuticle, and endocuticle in the head, thoracic, abdomen, and leg regions. Apparently, only skeletal muscles possessed muscle fibers in the muscular system. The main organ of the urinary system was the Malpighian tubule, the epithelium of which was lined with the simple cuboidal layer. The digestive system consisted of several distinct parts including foregut (for example esophagus, crop, and proventriculus), midgut, and hindgut (or rectum). Three different cell types were identified in the epithelial midgut. The respiratory organ was visible among the adipose tissue close to the integument system. Although this organ was not easily identified, it was covered by a simple squamous epithelium. The female reproductive system of this insect was a telotrophic meroistic ovary. The distribution of nurse cells was restricted to the anterior tropharium, connected to oocytes. All the above results provide the first histological description of X. compactus, which might be useful for advancing research in the related fields.

The structure and cell types of the organs of Halobates hayanus from Libong Island, Thailand, were investigated using a histological method. The integumentary system of this species consists of three layers: epicuticle, exocuticle, and endocuticle. Throughout the body, the muscular system exclusively included skeletal muscle. In the excretory system, we found four fully formed Malpighian tubules, each one lined with a straightforward cuboidal epithelium harbouring secretory granules. The digestive system comprised a foregut, midgut, and hindgut. The midgut had three distinct cell types: basal cells, epithelium, and absorptive cells. The nervous system comprised two regions: A ventral nerve cord and a frontal ganglion, which is connected to the eye structure. The frontal ganglion was composed of two lobes each containing an outer cortex and inner medulla. The outer cortex presented neurosecretory cells, neuroglia cells, and neurons. The neurosecretory cells were large and contained secretory granules in their cytoplasm. This histological study also showed the reproductive system of this gerrid species, including the reproductive tract, and the accessory organ.

This research aimed to investigate the effects of the intramuscular injection of vitamins AD3E and C in combination immediately before the estrus synchronization program (the Ovsynch program) on conception and pregnancy rates, blood parameters, serum biochemical properties, immune systems, antioxidant parameters, and proteomic and transcriptomic analyses during early gestation in dairy cows. Forty nonlactating multiparous cows were randomly assigned to one of four treatments: (1) C: control with normal saline injection; (2) VAD3E: a single intramuscular injection (I/M) of vitamin AD3E; (3) VAD3EC: injection of both vitamins AD3E and C; (4) VC: a single dose of vitamin C. Blood and serum samples were taken immediately at day 0 (before AI), day 7, and day 14 (after AI for 5 days) from the coccygeal vein. Generally, injections of AD3E and C in combination had no effect on the rate of conception or pregnancy. However, they improved hematological parameters and immune and antioxidant activities. Serum samples were analyzed using LC-MS/MS, and 8190 proteins were identified. Five proteins were successfully validated using the quantitative real-time reverse transcription PCR (qRT-PCR) method. This study found that lymphocyte-specific protein 1 (LSP1, A0A3Q1M894) could be used as a protein biomarker for cows administrated with vitamins AD3E and C.

The objective of this study was to describe the structure and histochemistry of the systemic organs in the female marine water-strider Asclepios annandalei from Pranburi river estuary, Thailand. Results from this study revealed for the first time that the integumentary system of this species consisted of three layers including epicuticle, exocuticle and endocuticle. The muscular system apparently contained only skeletal muscle along the body. In the urinary system, we observed well-developed Malpighian tubules, each of which was covered with the simple cuboidal epitheliums. These epitheliums also contained the secretory granules that were reacted positively with periodic acid Schiff (PAS). The digestive system of this species was composed of three distinct parts including foregut, midgut and hindgut. The respiratory system was composed of the respiratory organ, which was rarely found near the integument system. This organ was lined with a simple squamous epithelium. Two regions of nervous system, i.e., frontal ganglion connected to the eye structure and ventral nerve cord, were found. Each ganglion basically consisted of two layers, outer cortex and inner medullae. The outer cortex contained three types of cells, including neurosecretory cells, neuroglial cells and neurons. The cytoplasmic inclusion of neurosecretory cells contained secretory granules, which reacted positively with PAS, indicating the presence the glycoprotein. The neuroglia and neuron were also observed in the inner medullae layer. The female reproductive system (the ovarian structure, the reproductive tract and the accessory organ) of this gerrid species was seen under histological sections. The well-developed integument system and Malpighian tubule as well as the abundant respiratory organ is a characteristic of this species, which might be useful for the adaption to the estuarine condition.

BACKGROUND:Cannabinoids; tetrahydrocannabinol (THC), cannabidiol (CBD) and cannabinol (CBN), might show antibacterial activity. Trema orientalis is a species in the Cannabaceae that is closely related to Cannabis through plastome phylogenetic evidence. This species is widely distributed throughout tropical Asia and is used as traditional medicine, particularly for the treatment of infectious diseases. However, no studies on the antibacterial activity of cannabinoid-containing inflorescences extracts are available. Thus, the aim of this study was to determine cannabinoid content and antibacterial activity of inflorescences fractions from T. orientalis native to Thailand.METHODS:We hypothesized that inflorescences from T. orientalis might display cannabinoids similar to Cannabis because of their close taxonomic relationship. We extracted the mature inflorescences and infructescence of T. orientalis in three disparate populations from different Thailand floristic regions. Extractions were subsequently partitioned into hydrophilic and lipophilic fractions using distilled water and chloroform. The lipophilic extracts were further fractionated by the column chromatography with gradient elution and analyzed by gas chromatography-mass spectrometry (GC-MS). Characterized cannabinoids were used in bioassays with multidrug-resistance bacteria.RESULTS:Lipophilic extracts and fractions of inflorescences from all Thailand floristic regions consistently displayed cannabinoids (THC, CBD and CBN) in various quantities. These extracts exhibited inhibitory activity for Staphylococcus aureus, Pseudomonas aeruginosa, and Acinetobacter baumannii strains with minimum inhibitory concentration values varying from 31.25 to 125 µg/mL.CONCLUSION:Our study is the first to report cannabinoid detection in extracts from inflorescences of T. orientalis, a species in the Cannabaceae. These extracts and their fractions containing cannabinoids showed pronounced antibacterial activity. The use of analytic methods also demonstrated reproducible cannabinoid extraction.