Subterranean Termite Coptotermes Formosanus Research Articles

Even- and odd-numbered fatty acids used as worker arrestant aggregation pheromone components in the Formosan subterranean termite Coptotermes formosanus

Even- and odd-numbered fatty acids used as worker arrestant aggregation pheromone components in the Formosan subterranean termite Coptotermes formosanus

The influence of first desaturase subfamily genes on fatty acid synthesis, desiccation tolerance and inter-caste nutrient transfer in the termite Coptotermes formosanus.

Desaturase enzymes play an essential role in the biosynthesis of unsaturated fatty acids (UFAs). In this study, we identified seven "first desaturase" subfamily genes (Cfor-desatA1, Cfor-desatA2-a, Cfor-desatA2-b, Cfor-desatB-a, Cfor-desatB-b, Cfor-desatD and Cfor-desatE) from the Formosan subterranean termite Coptotermes formosanus. These desaturases were highly expressed in the cuticle and fat body of C. formosanus. Inhibition of either the Cfor-desatA2-a or Cfor-desatA2-b gene resulted in a significant decrease in the contents of fatty acids (C16:0, C18:0, C18:1 and C18:2) in worker castes. Moreover, we observed that inhibition of most of desaturase genes identified in this study had a negative impact on the survival rate and desiccation tolerance of workers. Interestingly, when normal soldiers were reared together with dsCfor-desatA2-b-treated workers, they exhibited higher mortality, suggesting that desaturase had an impact on trophallaxis among C. formosanus castes. Our findings shed light on the novel roles of desaturase family genes in the eusocial termite C. formosanus.

α-Terpineol affects social immunity, increasing the pathogenicity of entomopathogenic nematodes to subterranean termites (Isoptera)

Biocontrol of subterranean termites is largely impeded by their social immune responses. Studies on biocontrol agents combined with natural insecticides and their possible effects on the immune defense mechanisms of termites are limited. In this study, we investigated the effects of a combined biocontrol strategy using a plant-derived insect ATPase inhibitor, α-terpineol, with the entomopathogenic nematodes (EPNs) Steinernema carpocapsae against the subterranean termite Coptotermes formosanus Shiraki. Survival assays showed that even a low lethal concentration of α-terpineol significantly increased the EPNs-induced virulence in C. formosanus. α-terpineol treatment majorly inhibited the activity of Na+- K+- ATPase, which disturbed the EPNs-induced enhancement of locomotor activity and grooming behavior in termites treated with the combined strategy. Furthermore, the combination treatment had a synergistic inhibitory effect on innate immune responses in C. formosanus, which were measured as changes in the expression of immune-related genes and activities of immune system enzymes. In conclusion, α-terpineol can weaken the immune defense of termites against EPNs at low lethal concentrations, and is a suitable non-synthetic insecticide to prove the biocontrol efficiency of EPNs on C. formosanus. This study provides a theoretical basis and technical reference for a novel biocontrol strategy that promises to overcome the problems of host immune defense in termites.

Screening of reference genes for microRNA analysis in the study of solider caste differentiation of Formosan subterranean termite Coptotermes formosanus Shiraki

The soldier caste differentiation is a complex process that is governed by the transcriptional regulation and post-transcriptional regulation. microRNAs (miRNAs) are noncoding RNAs that control a wide range of activities. However, their roles in solider caste differentiation are barely studied. RT-qPCR is a powerful tool to study the function of genes. A reference gene is required for normalization for the the relative quantification method. However, no reference gene is available for miRNA quantification in the study of solider caste differentiation of Coptotermes formosanus Shiraki. In this research, in order to screen the suitable reference genes for the study of the roles of miRNAs in solider caste differentiation, the expression levels of 8 candidate miRNA genes were quantified in the head and thorax + abdomen during soldier differentiation. The qPCR data were analyzed using geNorm, NormFinder, BestKeeper, ΔCt method and RefFinder. The normalization effect of the reference genes was evaluated using the let-7-3p. Our study showed that novel-m0649-3p was the most stable reference gene, while U6 was the least stable reference gene. Our study has selected the most stable reference gene, and has paved the way for functional analysis of miRNAs in solider caste differentiation.

ALIGNMENT DAN ANALISIS KEKERABATAN RAYAP COPTOTERMES FORMOSANUS (Isoptera: Rhinotermitidae) PADA HABITAT YANG BERBEDA DENGAN SOFTWARE BIOEDIT DAN MEGA 6 (STUDI PUSTAKA DNA SEKUENSING DARI NCBI)

Background: Bioinformatics is a science that can relate information which includes molecular biology, biochemical structure, enzymatic, cell biology, physiology and pathology by using a computerized system based on data that has been collected. Methods: sequence alignment and phylogenetic tree construction of DNA sequences registered at NCBI in the subterranean termite Coptotermes formosanus Shiraki that lives in different habitats, namely in Mississippi, Southeast Asia, and China using Bioedit and MEGA software. Results: Sequence alignment showed that termite C. formosanus isolated from MS12 gene COII (FJ870577.1) in the Mississippi area had a base pair of 790 bp, termite C. formosanus isolated from Dinghu Mountain gene COII (KU257993.1) in China has a base pair of 779 bp, and the termite C. formosanus isolated from CF003JP gene COII (EF379943.1) in Southeast Asia has a base pair of 676 bp. The construction of the phylogenetic tree shows that the termite C. formosanus isolated from Dinghu Mountain gene COII (KU257993.1) in the China region has a close kinship with the termite C. formosanus isolated from CF003JP gene COII (EF379943.1) in the Southeast Asia region, while The termite C. formosanus isolated from MS12 gene COII (FJ870577.1) in the Mississippi area is distantly related to C. formosanus (KU257993.1) from China and C. formosanus (EF379943.1) from Southeast Asia. Conclusion: Mississippi has a base pair of 790 bp, termite C. formosanus isolated from Dinghu Mountain gene COII (KU257993.1), China has a base pair of 779 bp, and termite C. formosanus isolated from CF003JP gene COII (EF379943.1), Southeast Asia has a base pair of 676 bp. China is closely related to the termite C. formosanus isolated from COII gene CF003JP (EF379943.1), Southeast Asia, while the termite C. formosanus isolated from MS12 gene COII (FJ870577.1), Mississippi is distantly related to C. formosanus (KU257993.1) from China and C. formosanus (EF379943.1) from Southeast Asia.

Short Communication: Attractiveness of subterranean termite Coptotermes formosanus to plant leaf extracts

Abstract. Indrayani Y, Muin M, Adilla C, Yoshimura T. 2018. Short Communication: Attractiveness of subterranean termite Coptotermes formosanus to plant leaf extracts. Biodiversitas 19: 1176-1180. Plant-derived compounds have many potential uses. One such use is in agriculture for managing pests with less risk than with synthetic compounds that are toxicologically and environmentally undesirable. The current study focuses on the potential use of leaf extracts from five tropical plant species in attractant bait systems for controlling subterranean termites. Leaves from clove [Syzygium aromaticum (L.)], cajuput (Melaleuca leucadendra L.), cinnamon [Cinnamomum burmannii (Nees & T. Nees) Blume], basil (Ocimum sanctum L.) and bay (Syzygium polyanthum Wight) were extracted with 70% ethanol solution. Each extract was evaluated for its attractant properties for Coptotermes formosanus Shiraki using olfactory and Y-line trail following tests, respectively. Extract solutions were diluted to 1% concentration based on their eugenol content. For each test, which was replicated 15 times, a single worker was introduced into the test unit. The results showed that M. leucadendra crude extract was the most attractive among the five plant crude extracts, and the S. aromaticum crude extract was attractive especially in the trail-following test. The attraction rates were 40.00%, 80.00%, 53.33%, 60.00% and 46.67% for the S. aromaticum, M. leucadendra, C. burmannii, O. sanctum and S. polyanthum extracts, respectively. The trail-following rates were 86.67% for the S. aromaticum extract, 66.67% for the M. leucadendra extract, 53.33% for the C. burmannii extract, 20% for the O. sanctum extract as well as 26.67% for the S. polyanthum extract.

Age-based spatial distribution of workers is resilient to worker loss in a subterranean termite

Elaborate task allocation is key to the ecological success of eusocial insects. Termite colonies are known for exhibiting age polyethism, with older instars more likely to depart the reproductive center to access food. However, it remains unknown how termites retain this spatial structure against external disturbances. Here we show that a subterranean termite Coptotermes formosanus Shiraki combines age polyethism and behavioral flexibility to maintain a constant worker proportion at the food area. Since this termite inhabits multiple wood pieces by connecting them through underground tunnels, disastrous colony splitting events can result in the loss of colony members. We simulated this via weekly removal of all individuals at the food area. Our results showed that termites maintained a worker proportion of ~ 20% at the food area regardless of changes in total colony size and demographic composition, where younger workers replaced food acquisition functions to maintain a constant worker proportion at the food area. Food consumption analysis revealed that the per-capita food consumption rate decreased with younger workers, but the colony did not compensate for the deficiency by increasing the proportion of workers at the feeding site. These results suggest that termite colonies prioritize risk management of colony fragmentation while maintaining suitable food acquisition efficiency with the next available workers in the colony, highlighting the importance of task allocation for colony resiliency under fluctuating environments.

Colony Suppression and Possible Colony Elimination of the Subterranean Termites Coptotermes formosanus and Reticulitermes speratus by Discontinuous Soil Treatment Using a Diluent of Fipronil Suspension Concentrate.

Simple SummaryTermites play an important role in maintaining ecosystems, but they are also pests, exerting major economic impacts. Among the over 3000 known termite species, Coptotermes species and Reticulitermes species are the most common pest species. As one of the primary methods for controlling these subterranean termites, liquid termiticide is applied to the soil under and next to the building foundation to create a continuous chemical barrier. Nonrepellent slow-acting liquid termiticides, such as fipronil and imidacloprid, are used as the active ingredients in termite soil treatments. In the present study, to minimize the use of insecticides, a discontinuous soil treatment using fipronil was applied against subterranean termites, namely Coptotermes formosanus and Reticulitermes speratus, instead of the traditional continuous chemical barrier of termiticides. C. formosanus and R. speratus colonies were subjected to discontinuous soil treatments with fipronil and were strongly affected by the treatment at the colony level, resulting in colony suppression and possible colony elimination. Termite activity in the treated colony of C. formosanus was not found for more than two years, while that of R. speratus was not detected for three years.We assessed the efficacy of a discontinuous soil treatment using a diluent of fipronil suspension concentrate in controlling colonies of Coptotermes formosanus and Reticulitermes speratus. In-ground monitoring stations were installed at Isogi Park and Kindai University, and individual termites inhabiting the stations were collected for four or six years to determine the numbers and locations of colonies present in test areas before and after the discontinuous soil treatment. Microsatellite genotyping indicated that two C. formosanus and two R. speratus colonies in the test area at Isogi Park and five R. speratus colonies in the test area at Kindai University were active and that their territories fluctuated every year. One of the two C. formosanus colonies at Isogi Park and one of the five R. speratus colonies at Kindai University were subjected to discontinuous soil treatments with fipronil and were strongly affected by the treatment at the colony level, resulting in the suppression and possible elimination of colonies. Termite activity of the fipronil-treated colony of C. formosanus was detected within one week after the discontinuous soil treatment and was not found for more than two years (28 months), while termite activity of the fipronil-treated colony of R. speratus was detected within four days and three weeks after the discontinuous soil treatment and was not detected thereafter for three years. Fipronil residue analysis showed that workers of C. formosanus moved at least 28 m and that workers of R. speratus moved 6 m from the treated soil locations for up to three weeks.

Bridgehead effect and multiple introductions shape the global invasion history of a termite

Native to eastern Asia, the Formosan subterranean termite Coptotermes formosanus (Shiraki) is recognized as one of the 100 worst invasive pests in the world, with established populations in Japan, Hawaii and the southeastern United States. Despite its importance, the native source(s) of C. formosanus introductions and their invasive pathway out of Asia remain elusive. Using ~22,000 SNPs, we retraced the invasion history of this species through approximate Bayesian computation and assessed the consequences of the invasion on its genetic patterns and demography. We show a complex invasion history, where an initial introduction to Hawaii resulted from two distinct introduction events from eastern Asia and the Hong Kong region. The admixed Hawaiian population subsequently served as the source, through a bridgehead, for one introduction to the southeastern US. A separate introduction event from southcentral China subsequently occurred in Florida showing admixture with the first introduction. Overall, these findings further reinforce the pivotal role of bridgeheads in shaping species distributions in the Anthropocene and illustrate that the global distribution of C. formosanus has been shaped by multiple introductions out of China, which may have prevented and possibly reversed the loss of genetic diversity within its invasive range.

Cloning, Expression, and Immunological Characterization of Formosan Subterranean Termite (Blattodea: Rhinotermitidae) Arginine Kinase.

Several parts of the world regularly consume termites. Arthropod arginine kinase proteins often cross-react with human immunoblobulin E (IgE) antibodies and they are considered pan-allergens. The Formosan subterranean termite Coptotermes formosanus (C. formosanus (Shiraki) [Isoptera: Rhinotermitidae]), along with cockroaches, belong to the order Blattodea and they are common household pests in tropical and subtropical parts of the world. An sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) band migrating at approximately 37 kDa in C. formosanus termite extracts cross-reacted with IgE from five cockroach allergic patient samples by immunoblot. Liquid chromatography-mass spectrometry analysis of gel slices from the corresponding region of a gel indicated several peptides from the excised region were identical to the American cockroach arginine kinase allergen, Per a 9. The sequence of the full-length C. formosanus arginine kinase gene indicates the protein it encodes is 96% identical to American cockroach Per a 9, 94% identical to German cockroach Bla g 9, and 82-84% identical to shrimp arginine kinase proteins Pen m 2, Lit v 2, and Cra c 2. Full-length C. formosanus arginine kinase was fused to a glutathione S-transferase tag and recombinantly expressed and purified from Escherichia coli by affinity chromatography. The recombinant protein was recognized by IgE from 11 of 12 cockroach or shrimp allergic samples, but did not cross-react with dust mite allergic or peanut/tree nut allergic samples. The results of this study indicate the C. formosanus arginine kinase cross-reacts with cockroach and shrimp allergic IgE, and if consumed would likely act as an allergen.

Discovery of termitophilous rove beetles associated with Formosan subterranean termite Coptotermes formosanus in Taiwan, with the first larval description for the tribe Termitohospitini (Coleoptera: Staphylinidae)

The Formosan subterranean termite, Coptotermes formosanus Shiraki, 1909 (Blattodea: Rhinotermitidae), is a well-known pest of wood structures. Its native range has been widely investigated for potential implications for pest control strategies. Termitophilous staphylinid species of the genera Japanophilus Maruyama & Iwata, 2002 and Sinophilus Kistner, 1985 found in nests of C. formosanus have been used to infer the native range of their host termite. Herein, we extend the range of these two genera to include Taiwan, the type locality of C. formosanus. Taiwanese specimens were identified as J. hojoi Maruyama & Iwata, 2002 and S. yukoae Maruyama & Iwata, 2002 based on morphological and molecular data. The first-instar larva of J. hojoi is described, the first known immature stage of the tribe Termitohospitini (Coleoptera: Staphylinidae: Aleocharinae).

Screening of reference genes for expression analysis in the study of soldier caste differentiation of Formosan subterranean termite Coptotermes formosanus Shiraki.

The Formosan subterranean termite, Coptotermes formosanus Shiraki, is a serious pest worldwide. Juvenile hormone analog (and its analogs such as methoprene) can induce the transformation of the worker caste into soldier caste in C. formosanus. However, several factors, such as feeding substrate and colony origin, influence the proportion of solider formation. The molecular mechanism of worker to soldier transformation of C. formosanus is still not clear. RT-qPCR is a powerful tool for molecular studies. Accurate gene quantification by the relative quantification method requires a stable expressed gene as the reference gene. However, no reference genes were available for this species in the methoprene bioassay. To study the problem of gene response to methoprene by RT-qPCR we have to first screen reference genes in C. formosanus. Workers were fed with methoprene. Termites were collected during the methoprene bioassay and separated into head and thorax+abdomen. Expression profiles of 10 candidate reference genes in the two body part types were investigated using RT-qPCR. The results were analyzed by a set of established methods (geNorm, NormFinder, BestKeeper, and RefFinder) as well as comparative ΔCt method. Our results suggest that RPS18 is the most stably-expressed gene both in the head and thorax+abdomen for expression analysis in the methoprene bioassay of C. formosanus. The screening of suitable reference genes in C. formosanus establishes the foundation for the molecular study of soldier caste differentiation in this species.

The Termite Fecal Nest: A Framework for the Opportunistic Acquisition of Beneficial Soil Streptomyces (Actinomycetales: Streptomycetaceae).

Mutualistic associations between insects and microorganisms must imply gains for both partners, and the emphasis has mostly focused on coevolved host-symbiont systems. However, some insect hosts may have evolved traits that allow for various means of association with opportunistic microbial communities, especially when the microbes are omnipresent in their environment. It was previously shown that colonies of the subterranean termite Coptotermes formosanus Shiraki (Blattodea: Rhinotermitidae) build nests out of fecal material that host a community of Streptomyces Waksman and Henrici (Actinomycetales: Streptomycetaceae). These Actinobacteria produce an array of bioactive metabolites that provides a level of protection for termites against certain entomopathogenic fungi. How C. formosanus acquires and maintains this association remains unknown. This study shows that the majority of Streptomyces isolates found in field termite fecal nest materials are identical to Streptomyces isolates from soils surrounding the nests and are not vertically inherited. A survey of Streptomyces communities from C. formosanus fecal nest materials sampled at 20 locations around the world revealed that all nests are reliably associated with a diverse Streptomyces community. The C. formosanus fecal nest material therefore provides a nutritional framework that can recruit beneficial Streptomyces from the soil environment, in the absence of long-term coevolutionary processes. A diverse Streptomyces community is reliably present in soils, and subterranean termite colonies can acquire such facultative symbionts each social cycle into their fecal nest. This association probably emerged as an exaptation from the existing termite nest structure and benefits both the termite and the opportunistic colonizing bacteria.

Isolation and characterization of Metarhizium anisopliae TK29 and its mycoinsecticide effects against subterranean termite Coptotermes formosanus

The entomopathogenic fungus Metarhizium anisopliae is widely used as biocontrol agent against many insect pests. In the present study, the potential isolate of M. anisopliae TK29 was isolated from the agricultural soils in Thekkady, India. The taxonomic identity of the isolate was confirmed based on its morphology and 18S rDNA gene sequence homology. Phylogenetic analysis confirmed that the isolated strains were related to the same species. A potential isolate (TK29) was optimized for mass cultivation and conidial spore production was enhanced using three different raw substrates (Rice, Maize, black gram) by solid-state fermentation. The results showed higher conidial spore yield from rice (2.6 ± 0.32%) compared to black gram (2.1 ± 0.28%) and maize (1.9 ± 0.23%) substrates. Dry green conidia were applied against Formosan subterranean termite, Coptotermes formosanus at three different concentrations (1 × 106, 1 × 107, and 1 × 108 conidia/ml−1). The highest mortality rate was obtained from 1 × 108 conidia/ml−1 at 120 h post-treatment. Our study indicated that M. anisopliae TK29 had desirable attributes for the development of a mycoinsecticide against C. formosanus.

Chemical Interaction among Termite-Associated Microbes

Bacteria and fungi in shared environments compete with one another for common substrates, and this competition typically involves microbially-produced small molecules. An investigation of one shared environmental niche, the carton material of the Formosan subterranean termite Coptotermes formosanus, identified the participants on one of these molecular exchanges. Molecular characterization of several termite-associated actinobacteria strains identified eleven known antimicrobial metabolites that may aid in protecting the C. formosanus colony from pathogenic fungal infections. One particular actinobacterial-derived small molecule, bafilomycin C1, elicited a strong chemical response from Trichoderma harzianum, a common soil saprophyte. Upon purification and structure elucidation, three major fungal metabolites were identified, t22-azaphilone, cryptenol, and homodimericin A. Both t22-azaphilone and homodimericin A are strongly upregulated, 123- and 38-fold, respectively, when exposed to bafilomycin C1, suggesting each play a role in defending T. harzianum from the toxic effect of bafilomycin C1.

Efficacy of 1% fipronil dust of activated carbon against subterranean termite Coptotermes formosanus Shiraki in laboratory conditions

Toxicity and horizontal transmission of 1% fipronil dust of activated carbon were measured using the subterranean termite Coptotermes formosanus Shiraki in laboratory conditions. 1% fipronil dust of activated carbon has delayed toxicity towards C. formosanus compared with 0.5% fipronil dust of French chalk; knockdown times KT50 and KT90 were delayed by >9 and >15 h respectively. Furthermore, 1% fipronil dust of activated carbon showed excellent primary and secondary horizontal transfer levels. In primary horizontal transfer, recipient mortalities reached 100% by 24, 48 and 72 h at donor-recipient ratios of 1:1, 1:5 and 1:10, respectively. High transfer efficacies were also found if donor-recipient ratios were greatly increased: mortality reached 100% at 9 d at ratio 1:25 and >90% at 12 d at 1:50. In secondary horizontal transfer, the toxicant transmitting ability of C. formosanus was greater when the primary horizontal transfer ratio was lower, and the highest transfer efficacy was found with a donor-recipient ratio of 1:1 - recipient mortalities reached 100% at 5 d and 11 d, respectively. Application of 1% fipronil dust of activated carbon overcomes the problem that that too high a concentration kills termites before they can contaminate their nestmates, while a lower concentration may not supply a sufficient dose for effective transfer from treated to untreated termites; this preparation has delayed toxicity, dose-dependent toxicity in horizontal transfer and high efficacy to control C. formosanus.

Toxicity and Feeding Deterrent Effect of 2-Methylanthraquinone from the Wood Extractives of Tectona grandis on the Subterranean Termites Coptotermes formosanus and Reticulitermes speratus

No-choice feeding tests using ethanol, chloroform, and acetone extractives of teak (Tectona grandis) heartwood clearly showed feeding deterrent activity and toxicity to the subterranean termite Reticulitermes speratus. The amount of 2-methylanthraquinone (MAQ) in teak wood extractives was not related to the feeding deterrents or toxicity, as shown by the no-choice feeding tests conducted using crude extractives containing various amounts of MAQ, MAQ alone, and fractions of crude extractives. As a native pest, the subterranean termite Coptotermes formosanus was more tolerant to the fractions of crude extractives than Reticulitermes speratus, and the mortality observed in C. formosanus was not due to the presence of MAQ.

Crude extracts of two different leaf plant species and their responses against subterranean termite Coptotermes formosanus

Abstract. Indrayani Y, Muin M, Yoshimura T. 2016. Crude extracts of two different leaf plant species and their responses against subterranean termite Coptotermes formosanus. Nusantara Bioscience 8: 226-231. Crude extracts of plants consist of various chemical compounds useful for many purposes and their effects depend on the comprising active ingredients and applied concentrations. This study was conducted to determine the chemical compounds of plant leaf extracts and examine their responses against subterranean termite Coptotermes formosanus Shiraki. Two tropical plant species [clove (Syzygium aromaticum) and cajuput (Melaleuca leucadendra)] were extracted with 70% ethanol solvent. As the two different plant crude extracts contain eugenol, though to have termiticidal performance, various concentrations (0.1%, 0.05%, and 0.01%) based on their eugenol contents were prepared with filter paper samples for the test. The termite mortality and mass loss of extract-treated and untreated control filter paper samples were determined after four weeks of exposure. Results showed that crude extracts of different plant species showed different responses against the subterranean termites although they were applied at the same eugenol-based concentration. At the concentration of 0.1%, the crude extract of S. aromaticum caused termite mortality and consumption rates of 100% and 0.3%, respectively. Whilst, the crude extract of M. leucadendra caused significantly lower termite mortality (36.8%) and higher consumption (9.1%). Samples with lower eugenol-based concentration showed lower termite mortality and may have attractive effects on termites as indicated by higher consumption at the concentration of 0.01% compared to untreated controls. These results suggested that crude extracts of different two leaf species had different effects on subterranean termite C. formosanus even though they were applied at the same eugenol-based concentrations.

Effect of Temperature and Humidity on Survival of Coptotermes formosanus and Reticulitermes flavipes (Isoptera: Rhinotermitidae)

Two subterranean termite species were subjected to combinations of six temperatures (10°, 15°, 20°, 25°, 30°, or 35°C) and five relative humidities (RH) (55, 65, 75, 85, or 99%) to determine optimum conditions for survival. When small groups of the Formosan subterranean termite Coptotermes formosanus Shiraki or the eastern subterranean termite Reticulitermes flavipes (Kollar) were exposed to all 30 combinations of temperature and RH, survival times were significantly influenced by temperature, RH, and their interaction. For both species, survival times were longest at low temperatures and high RH. Maximum survival of small groups of C. formosanus and R. flavipes workers and soldiers occurred at the combination of 10°C and 99% RH C (LT50= 28.2 d, LT50 = 18.1 d, respectively). Survival of paired C. formosanus dealates was evaluated at combinations of 20°, 25°, or 30°C and 55, 65, 75, 85, or 99% RH. Survival was strongly influenced by temperature and humidity. Longest survival times until 50% mortality occurred at 99% RH and 20° or 25°C (LT50= 2.5 d, LT50 = 3.0 d, respectively). At all temperatures, mortality occurred too quickly for LT50 values to be determined when RH was 55% or 65%.

Molecular Diagnostic Technique for the Differentiation of the Formosan Subterranean Termite,Coptotermes formosanus(Isoptera: Rhinotermitidae) from Other Subterranean Termites by Multiplex-PCR

The Formosan subterranean termite Coptotermes formosanus Shriaki; (Isoptera: Rhinotermitidae), is a major pest that is spreading throughout the southeastern United States. Morphological identification of worker specimens is not possible using available taxonomic keys based on morphological traits. A multiplex PCR protocol was developed that can differentiate the Formosan subterranean termite from other termite species in a single PCR reaction. This multiplex PCR protocol simplifies previous molecular diagnostic techniques.