Hybrids of two destructive subterranean termites established in the field, revealing a potential for gene flow between species.

Hybridization between invasive pest species may lead to significant genetic and economic impacts that require close monitoring. The two most invasive and destructive termite species worldwide, Coptotermes formosanus Shiraki and Coptotermes gestroi (Wasmann), have the potential for hybridization in the field. A three-year field survey conducted during the dispersal flight season of Coptotermes in Taiwan identified alates with atypical morphology, which were confirmed as hybrids of the two Coptotermes species using microsatellite and mitochondrial analyses. Out of 27,601 alates collected over three years, 4.4% were confirmed as hybrid alates, and some advanced hybrids (>F1 generations) were identified. The hybrid alates had a dispersal flight season that overlapped with the two parental species 13 out of 15 times. Most of the hybrid alates were females, implying that mating opportunities beyond F1 may primarily be possible through female hybrids. However, the incipient colony growth results from all potential mating combinations suggest that only backcross colonies with hybrid males could sometimes lead to brood development. The observed asymmetrical viability and fertility of hybrid alates may critically reduce the probability of advanced-hybrid colonies being established in the field.

The Trail-Following Communication in Stylotermes faveolus and S. halumicus (Blattodea, Isoptera, Stylotermitidae).

Stylotermitidae appear peculiar among all termites, feeding in trunks of living trees in South Asia only. The difficulty to collect them limits the ability to study them, and they thus still belong to critically unknown groups in respect to their biology. We used a combination of microscopic observations, chemical analysis and behavioural tests, to determine the source and chemical nature of the trail-following pheromone of Stylotermes faveolus from India and S. halumicus from Taiwan. The sternal gland located at the 5th abdominal segment was the exclusive source of the trail-following pheromone in both S. faveolus and S. halumicus, and it is made up of class I, II and III secretory cells. Using gas chromatography coupled mass spectrometry, (3Z)-dodec-3-en-1-ol (DOE) was identified as the trail-following pheromone which elicits strong behavioural responses in workers at a threshold around 10- 4 ng/cm and 0.1 ng/gland. Our results confirm the switch from complex aldehyde trail-following pheromones occurring in the basal groups to simpler linear alcohols in the ancestor of Kalotermitidae and Neoisoptera.

Soldier phenotypic differences among 2 invasive and destructive Coptotermes species and their hybrids (Blattodea: Isoptera: Rhinotermitidae).

With recent evidence of hybridization events in the field, the phenotypic traits of F1 hybrid colonies of 2 destructive subterranean termite species, Coptotermes formosanus Shiraki and Coptotermes gestroi (Wasmann) remain to be investigated. In this study, laboratory colonies of 2 conspecific pairings and 2 heterospecific pairings (hybrid F = ♀C. formosanus × ♂C. gestroi, hybrid G = ♀C. gestroi × ♂C. formosanus) were examined in Florida, USA, and in Taiwan. Colony nest architecture for both hybrids displayed disorganized carton materials compared to the defined trabecular carton of both parental species. Soldier head measurements were not a reliable approach for diagnostic purposes, as soldier morphometric traits widely overlapped across all mating combinations, except for hybrid F soldiers displaying abnormally long mandibles. Hybrid F soldiers' mandibles also remained parallel when at rest. However, 4 qualitative morphological differences in soldiers were determined for diagnostic purposes. First, the fontanelle in both hybrids is horizontally ellipsoid whereas subcircular in C. gestroi and trianguliform in C. formosanus. Second, sclerotized striations along the postmental sulcus are present in C. gestroi, absent in C. formosanus, and intermediate in both hybrid soldier types. Third, each lateral margin of the fontanelle is flanked by 2 setae in C. formosanus and both hybrids, while a single seta resides on each side of the fontanelle in C. gestroi. Finally, C. gestroi and hybrid soldiers' heads are characterized by a bulging vertex that is lacking in C. formosanus. Therefore, a combination of these 4 characteristics now allows for soldier identification of hybrid Coptotermes.

Digesta retention time and recovery rates of ants and termites in Chinese pangolins (Manis pentadactyla).

Pangolins are myrmecophagous mammals whose biology and ecology remain poorly studied. Termite mandibles and ant head capsules are the two primary remains found in pangolin feces. Determining the retention time of insect cuticles is important for understanding the digestive physiology of pangolins, while determining the recovery rate of termites and ants in feces is required to estimate the number of these prey items that are consumed by pangolins. In this study, the authors conducted feeding trials with captive Chinese pangolins (Manis pentadactyla). Sixty grams of the fungus-growing termite Odontotermes formosanus (18,816 individuals) and 15-20 g of the yellow crazy ant Anoplolepis gracilipes (14,400-19,200 individuals) were fed to each pangolin. After feeding, pangolin feces were collected daily for 1 week. The authors also assessed the accuracy of using chromium (III) oxide powder (Cr2 O3 ) as a proxy for determining gut passage time, as has been done in previous studies. The results showed that remaining termite mandibles and ant head capsules in feces peaked at 66 and 90 hr after feeding and their recovery rates were 0.35 ± 0.10 and 0.65 ± 0.04, respectively. In both feeding trials, the retention time of Cr2 O3 was much shorter than that of the termite mandibles and ant head capsules, indicating that Cr2 O3 is not an appropriate indicator for estimating food retention time of myrmecophagous animals. Our results revealed that the ant head capsules were preserved better in feces compared with the termite mandibles, suggesting that termites may be considerably underestimated in the feces of wild pangolins.

Divergent effects of climatic factors on termite body size: alate versus worker castes.

Body size is an important functional trait to animals. Caste division of eusocial insects can exert a profound influence on their interactions with environment. We investigate the intra-specific variation of body size between caste within Odontotermes formosanus (Shiraki) (Blattodea: Termitidae), the most common and widely distributed termite species in Taiwan Island. By utilizing specimens from the NCHU Termite Collection and WorldClim data, we describe the body size distribution pattern of O. formosanus on two castes, worker and alate, and relationship with climatic factors is examined. The body size of workers is positively correlated with latitude and elevation. The body size of alates does not correlate with latitude but is positively correlated with elevation. Temperature factors negatively affect the body size of both castes. Precipitation has a positive effect on the body size of alates and no effect on workers. Additionally, humidity and temperature fluctuations over time have divergent effects on the body size of alates and workers. The results provide evidence of trait evolution decoupling at the intraspecific level, which may be shaped by climatic factors.

Three-layered functionally specialized nest structures enhance strength and water retention in mounds of Globitermes sulphureus (Blattodea: Termitidae).

Termite mounds are a ubiquitous feature of savanna ecosystems, yet the mechanisms by which termites (Blattodea: Termitoidae) mitigate the challenges posed by seasonal drought and flooding through mound construction remain insufficiently explored. This study investigates the material properties, water retention capabilities, and resistance to raindrop penetration across three distinct layers of Globitermes sulphureus (Haviland, 1898) (Blattodea: Termitidae) nests. Our findings reveal a pronounced diversification and specialization of materials and functions across these layers. Specifically, the outer layer has decreased moisture permeability, the middle layer has enhanced resistance to water penetration, and the innermost layer has a high capacity for water retention. The integration of these functionally specialized layers provides a compelling evidence for explaining how these termites are able to adapt to the fluctuating environmental conditions characteristic of savanna ecosystems. Furthermore, this highlights the nest's buffering capability against environmental stressors. The complexity of this construction, marked by a level of self-organization rarely observed in the animal kingdom, underscores a significant instance of architectural ingenuity among non-human builders.

Monoclonal neutralizing antibodies against SARS-COV-2 S protein.

Novel coronavirus pneumonia, also known as coronavirus disease 2019 (COVID-19), is caused by sub-severe acute respiratory syndrome type 2 coronavirus (SARS-CoV-2) infection. The spike (S) protein of SARS-CoV-2 binds to angiotensin-converting enzyme 2 (ACE2) receptors widely expressed on the surface of human cells leading to life-threatening respiratory infections. A serious hazard to human health is posed by the lack of particular treatment medications for this virus infection. We advocate the creation of high-affinity antibodies using the receptor binding domain (RBD) of S protein as a specific antigenic epitope to develop a drug that can precisely target therapy COVID-19 because SARS-CoV-2 infection of the host cells is dependent on S protein binding to ACE2. Finally, we obtained high-affinity antibodies 14F4HL and 14E3HL that have high affinity with RBD and well-drug-forming properties, suitable for further humanization studies. Thus, monoclonal antibodies that neutralize the S protein were identified in our study, which may provide new insights for the development of COVID-19 therapeutic drugs.

Defensive glands in Stylotermitidae (Blattodea, Isoptera).

The large abundance of termites is partially achieved by their defensive abilities. Stylotermitidae represented by a single extant genus, Stylotermes, is a member of a termite group Neoisoptera that encompasses 83% of termite species and 94% of termite genera and is characterized by the presence of the frontal gland. Within Neoisoptera, Stylotermitidae represents a species-poor sister lineage of all other groups. We studied the structure of the frontal, labral and labial glands in soldiers and workers of Stylotermes faveolus, and the composition of the frontal gland secretion in S. faveolus and Stylotermes halumicus. We show that the frontal gland is a small active secretory organ in soldiers and workers. It produces a cocktail of monoterpenes in soldiers, and some of these monoterpenes and unidentified proteins in workers. The labral and labial glands are developed similarly to other termite species and contribute to defensive activities (labral in both castes, labial in soldiers) or to the production of digestive enzymes (labial in workers). Our results support the importance of the frontal gland in the evolution of Neoisoptera. Toxic, irritating and detectable monoterpenes play defensive and pheromonal functions and are likely critical novelties contributing to the ecological success of these termites.

Genomic data provide insights into the classification of extant termites.

The higher classification of termites requires substantial revision as the Neoisoptera, the most diverse termite lineage, comprise many paraphyletic and polyphyletic higher taxa. Here, we produce an updated termite classification using genomic-scale analyses. We reconstruct phylogenies under diverse substitution models with ultraconserved elements analyzed as concatenated matrices or within the multi-species coalescence framework. Our classification is further supported by analyses controlling for rogue loci and taxa, and topological tests. We show that the Neoisoptera are composed of seven family-level monophyletic lineages, including the Heterotermitidae Froggatt, Psammotermitidae Holmgren, and Termitogetonidae Holmgren, raised from subfamilial rank. The species-rich Termitidae are composed of 18 subfamily-level monophyletic lineages, including the new subfamilies Crepititermitinae, Cylindrotermitinae, Forficulitermitinae, Neocapritermitinae, Protohamitermitinae, and Promirotermitinae; and the revived Amitermitinae Kemner, Microcerotermitinae Holmgren, and Mirocapritermitinae Kemner. Building an updated taxonomic classification on the foundation of unambiguously supported monophyletic lineages makes it highly resilient to potential destabilization caused by the future availability of novel phylogenetic markers and methods. The taxonomic stability is further guaranteed by the modularity of the new termite classification, designed to accommodate as-yet undescribed species with uncertain affinities to the herein delimited monophyletic lineages in the form of new families or subfamilies.

Predicting habitat suitability of Coptotermes gestroi (Isoptera: Rhinotermitidae) with species distribution models.

Coptotermes gestroi (Wasmann) is an important structural pest reported from Asia, Pacific islands, North America, Caribbean islands, South America, and Indian Ocean islands. This study summarized previous records of C. gestroi and its synonyms, presenting 184 infested counties from 24 countries. Based on the geo-references occurrence locations and global raster data of climate, geography, and human population, C. gestroi were found most commonly in warm, high precipitation, low altitude, and human populated areas. By using species distribution models, we predicted its current infested area (model 1), habitat suitability (model 2), and probability of introduction (model 3) on a global scale. The results showed its recorded locations and the predicted distribution of the present day are similar, but the suitable habitat is larger than its current distribution. The patterns of the introduction frequency (model 3) and habitat suitability (model 2) are inconsistent. Temperate cities with high introduction risk are located in Europe, United Sates, northeastern China, and Japan where habitat suitability is low and hence successful colonization is unlikely. In tropics and subtropics, habitat suitability of C. gestroi is high. We speculate that continuous urbanization and increasing human population will increase its introduction frequency and cause further extension in fast developing tropical and subtropical countries.

Desiccation tolerance of Termitidae termites in relation to their nest type.

The family Termitidae is renowned for its diverse nesting behaviors, with the evolution of epigeal and arboreal nests hypothesized to increase desiccation stress due to greater exposure to air. However, these nests may also alleviate desiccation stress through humidity regulation. To explore the implications of acquiring epigeal and arboreal nests, we investigated desiccation tolerance traits in 16 Termitidae termite species with varying nest types and analyzed trait correlations. Principal component analysis revealed that termites constructing epigeal and arboreal nests exhibited reduced water loss rates and enhanced survival under desiccated conditions. Furthermore, termites building arboreal nests displayed a notably higher water content. Redundancy analysis demonstrated that nest types accounted for a substantial portion (57.2%) of the observed variation in desiccation tolerance. These findings support the hypothesis that epigeal and arboreal nests in termites are associated with increased desiccation stress and increased desiccation tolerance. These findings highlight the role of nest type in influencing desiccation tolerance mechanisms and water regulation strategies in termites.

Body size of fungus-growing termites infers on the volume and density of their fungal cultivar.

The body size of an animal plays a crucial role in determining its trophic level and position within the food web, as well as its interactions with other species. In the symbiosis between Termitomyces and fungus-growing termites, termites rely on nutrition of fungal nodules produced by Termitomyces. To understand whether the size of termites and fungal nodules are related to their partner specificity, we quantified the size of termite farmer caste, and the size and density of nodules in termite nests of four genera of fungus-growing termites, and identified their cultivated Termitomyces fungus species based on internal transcribed spacer regions and partial large subunit ribosomal RNA gene sequences. The results showed that the size and density of fungal nodules were different among Termitomyces clades and revealed a constant trade-off between size and density among clades. The nodule size of each clade has low variation and fits normal distribution, indicating that size is a stabilized trait. Moreover, we found larger termite genera cultivated Termitomyces with larger but less numerous nodules. Based on these results, we concluded that there is a size specificity between Termitomyces and fungus-growing termites, which may lead to diversification of Termitomyces as adaptations to different termite genera.

Diet composition of an escaped captive-born southern tamandua (Tamandua tetradactyla) in a nonnative habitat in Asia.

Studies on the role of natural predatory instincts in captive-born mammalian myrmecophagy are rare. Consequently, researchers rely extensively on case reports to learn more about the contexts in which predatory behavior occurs among such animals. In this study, we recorded an uncommon case of a captive-born southern tamandua (Tamandua tetradactyla) that accidentally escaped from a zoo into a nonnative habitat in Asia. The southern tamandua was found alive 3 months later. Two fresh fecal samples were obtained, and the diet composition was examined. Three termite species (one family, three genera), and 14 ant species (four subfamilies, nine genera) were identified in the fecal samples. The studied southern tamandua preyed on terrestrial and arboreal ants and termites, as the wild populations of its species do. Ants of the subfamily Myrmicinae and termites of the subfamily Nasutitermitinae were the most abundant prey items in the samples, which is consistent with related reports on the wild populations. Soldier ants constituted <1% of the prey items in the fecal samples, suggesting that the southern tamandua likely avoided preying on ants of the soldier caste. Fungus-growing termites Odontotermes (Isoptera: Macrotermitinae), which are not native to neotropical regions, were also ingested by the southern tamandua. This study provides information on how a captive-born mammalian myrmecophagy applies its natural feeding instincts in nonnative natural settings.

Functional Plasticity of Foraging Shelter Tubes Built by Termites.

Social insects' constructions are diverse and functionally flexible. We aimed to understand how termites modify their behavior and building material to change the function of a shelter tube. Construction behaviors of three termite species, Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae), Odontotermes formosanus (Shiraki) (Blattodea: Termitidae), and Nasutitermes takasagoensis (Nawa) (Blattodea: Termitidae), were recorded and analyzed. We measured the thickness and organic matter content of shelter tubes, and further tested their water loss and water drop resistance. All three termite species used a cement-brick construction procedure. The shelter tubes of C. formosanus and N. takasagoensis incorporating of woody material and feces present a strong resistance to water drops. Shelter tubes of the fungus-growing termite, O. formosanus, constructed using only soil and fluid secretion had a weak resistance to water drops. The trade-off between using feces to increase hydrophobicity of shelter tubes and using feces to grow fungus likely occurs in fungus-growing termites. Among intraspecific colonies, increasing the thickness of shelter tube improved resistance to water loss and water drops. This study demonstrated termites employed limited construction material and building behaviors to achieve functional plasticity of foraging shelter.

Drugstore Beetle Damage Pattern of Glue Paste-Lined Oil Paintings.

A glue paste lining is a traditional conservation treatment used to reinforce the aged canvas of oil paintings. Several insect infestation cases concerning glue paste-lined oil paintings have been reported around the world, particularly in European countries. In 2008, Chimei Museum, a private museum located in Tainan, Taiwan, was affected by a severe beetle infestation of oil paintings. To confirm the infesting insects and to quantify the damage pattern for further development of control and monitoring methods, three severely damaged paintings were examined along with the restoration procedure. A total of four insect species were collected. The drugstore beetle, Stegobium paniceum (L.) (Coleopter: Ptinidae), was the primary pest found in all three paintings investigated and was identified based on morphological and genetic data. Thrips and booklice were considered secondary pests, and a Pteromalid wasp was speculated to have a parasitic relationship with the drugstore beetle. Drugstore beetle larvae mainly bored into the glue paste layer and original canvas and required only 5.94 ± 1.38 mm3 of feed to grow from egg to pupa. Their bores were not evenly distributed, and most of them were found in the shaded area covered by the stretcher and outer frame. The body length of drugstore beetles varied, ranging from 1.67 to 2.75 mm, which may explain the various sizes of exit holes on gummed tape surrounding the frame. Detailed and quantified information on drugstore beetle's pattern of damage provided in this study will be beneficial for further developing conservation practices and inspection methods.

Nationwide Termite Pest Survey Conducted in Taiwan as a Citizen Science Project.

Information regarding the species composition and dispersal flight season of termites is crucial for termite management. The major obstacles to collecting such information are a lack of access to private buildings and shortage of workers to monitor and report on termite swarming. To overcome these difficulties, we launched a citizen science project in which members of the public and pest management professionals were invited to collect termite samples. We created the website, Taiwan Termite Identification Service, on which populace could log the collection information, and ship termite samples to our laboratory for identification. We also established a Facebook group, called the "Termite Forum," to publicize this project. A total of 3024 samples were collected from 2015 to 2020, and we identified the species of >93% of the samples. Based on 1499 samples collected from buildings, five structural termite pests were identified, and species composition in each county of Taiwan is available. According to 844 dispersal flight events, termite dispersal flight timing peak and degree of centralization were estimated using a Gaussian model. The collected data demonstrated that the invasive termite species, Coptotermes gestroi (Wasmann) (Blattodea: Rhinotermitidae), continued northward expansion. The first intercepted alate of Schedorhinotermes sp. (Blattodea: Rhinotermitidae) indicated that it may be a new invasive pest from Southeast Asia. This study reports on a successful case of a citizen science project where urban pest data were collected on a national scale.

Area-population control of fungus-growing termite, Odontotermes formosanus, using hexaflumuron durable baits.

BACKGROUND: Fungus-growing termites (Termitidae: Macrotermitinae) are common forest and agriculture pests. To evaluate the efficacy of termite baiting in suppressing field population of fungus-growing termites, a durable termite bait with hexaflumuron was evenly installed in a one-hectare forest area dominated by a fungus-growing termite, Odontotermes formosanus (Shiraki). Monthly monitoring of termite foraging activity on baits and wood stakes was conducted for 4 years to quantify efficacy of baits. To examine whether the hexaflumuron led to colony death, pesticides in fungus gardens of active and deceased nests were quantified using a LC-QTOF/MS. RESULTS: After baiting, 50% and 90% of baits were fed upon 10 and 24 months, respectively. After 2 years of baiting, the monthly number of wood stakes occupied by termites was reduced from 34.7 ± 1.8 to 17.6 ± 2.5 (-49.1%), and the number of wood stakes consumed was reduced from 17.7 ± 0.8 to 13.3 ± 1.2 (-25.7%). Hexaflumuron was detected in deceased colonies, including five of six fungus gardens and the fungal tissue of Xyleria grown on fungus gardens, with a concentration of 0.31-20.11 mg kg-1 dry weight. CONCLUSION: This study demonstrated that durable hexaflumuron baits consumed by fungus-growing termites were further incorporated into fungus gardens, resulted in colony elimination and negative area-population effects, supporting that durable hexaflumuron baits are effective in suppressing field populations of fungus-growing termites. © 2021 Society of Chemical Industry.

Termite Pest Identification Method Based on Deep Convolution Neural Networks.

Several species of drywood termites, subterranean termites, and fungus-growing termites cause extensive economic losses annually worldwide. Because no universal method is available for controlling all termites, correct species identification is crucial for termite management. Despite deep neural network technologies' promising performance in pest recognition, a method for automatic termite recognition remains lacking. To develop an automated deep learning classifier for termite image recognition suitable for mobile applications, we used smartphones to acquire 18,000 original images each of four termite pest species: Kalotermitidae: Cryptotermes domesticus (Haviland); Rhinotermitidae: Coptotermes formosanus Shiraki and Reticulitermes flaviceps (Oshima); and Termitidae: Odontotermes formosanus (Shiraki). Each original image included multiple individuals, and we applied five image segmentation techniques for capturing individual termites. We used 24,000 individual-termite images (4 species × 2 castes × 3 groups × 1,000 images) for model development and testing. We implemented a termite classification system by using a deep learning-based model, MobileNetV2. Our models achieved high accuracy scores of 0.947, 0.946, and 0.929 for identifying soldiers, workers, and both castes, respectively, which is not significantly different from human expert performance. We further applied image augmentation techniques, including geometrical transformations and intensity transformations, to individual-termite images. The results revealed that the same classification accuracy can be achieved by using 1,000 augmented images derived from only 200 individual-termite images, thus facilitating further model development on the basis of many fewer original images. Our image-based identification system can enable the selection of termite control tools for pest management professionals or homeowners.

Termite's Twisted Mandible Presents Fast, Powerful, and Precise Strikes.

The asymmetric mandibles of termites are hypothetically more efficient, rapid, and powerful than the symmetric mandibles of snap-jaw ants or termites. We investigated the velocity, force, precision, and defensive performance of the asymmetric mandibular snaps of a termite species, Pericapritermes nitobei. Ultrahigh-speed recordings of termites revealed a new record in biological movement, with a peak linear velocity of 89.7-132.4 m/s within 8.68 µs after snapping, which caused an impact force of 105.8-156.2 mN. High-speed video recordings of ball-strike experiments on termites were analysed using the principle of energy conservation; the left mandibles precisely hit metal balls at the left-to-front side with a maximum linear velocity of 80.3 ± 15.9 m/s (44.0-107.7 m/s) and an impact force of 94.7 ± 18.8 mN (51.9-127.1 mN). In experimental fights between termites and ant predators, Pe. nitobei killed 90-100% of the generalist ants with a single snap and was less likely to harm specialist ponerine ants. Compared with other forms, the asymmetric snapping mandibles of Pe. nitobei required less elastic energy to achieve high velocity. Moreover, the ability of P. nitobei to strike its target at the front side is advantageous for defence in tunnels.

Identification of Subterranean Termite Genus, Reticulitermes (Blattodea: Rhinotermitidae) in Taiwan.

Species identification in the genus Reticulitermes is often difficult because of ambiguous morphological characters. Challenges in Reticulitermes spp. recognition have also been encountered in East Asia, including Taiwan. Because of unknown Reticulitermes taxa in Taiwan and the possible origin of alien Reticulitermes kanmonensis Takematsu in Japan and Korea, reexamining Reticulitermes fauna in Taiwan is imperative. To clarify the Reticulitermes fauna in Taiwan, this study applied two mitochondrial genes (cytochrome oxidase subunit II [COII] and 16S rDNA) and morphological characters for species delimitation. Reticulitermes specimens collected from 63 localities across the main and adjacent islands of Taiwan were analyzed. Phylogenetic analyses, morphological comparisons, and ecological traits suggested the existence of three species in Taiwan: Reticulitermes flaviceps (Oshima), R. kanmonensis, and Reticulitermes leptomandibularis Hsia and Fan. Altitudinal distributions among the three Reticulitermes termites tended to differ: R. flaviceps adapted to low hills, but R. kanmonensis and R. leptomandibularis occurred in medium mountainous areas. The combined data, including haplotype diversities and distribution range, suggest that 1) R. flaviceps is an endemic species and only found in Taiwan; 2) R. kanmonensis and R. leptomandibularis are both native species in Taiwan and China; 3) Japanese R. kanmonensis populations originated from southern China and/or Taiwan and that Korean populations were possibly introduced from Japan.