乳白蚁属Coptotermes(Isoptera: Rhinotermitidae)检疫分子鉴定研究进展

白蚁是一类受到世界广泛关注的重要社会性昆虫,对人类生活、经济以及生态环境等有很大的影响,特别是乳白蚁属Coptotermes众多种类危害尤为严重。乳白蚁常随木质包装或原木远距离传播异地建群,其为害隐蔽、潜伏期长,一旦爆发则为害严重且难防治,因其种类多、形态趋同,同一巢群中有龄期不一的虫态和多种生物型,可用分类特征少,造成形态分类困难、人为分类现象普遍。近年来分子生物学技术的日益发展,使得白蚁的分类鉴定更加快速、灵敏和可靠,系统发育分析研究更为深入全面,遗传进化方面取得大量研究成果。本文对基于生物信息学的乳白蚁属检疫分类鉴定和系统发育研究方法进行了介绍和评述,并对DNA技术在乳白蚁属分类和进化研究中的应用进行了讨论。     

Directional selection by the subterranean termite Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae) at tunnel intersections

Subterranean termites construct complicated tunnel networks for foraging below ground. Thus, they often encounter tunnel intersections during foraging activity. Directional selection by termites at intersections is likely to affect foraging efficiency because depending on their selection, the path length from food resources to the nest can vary significantly. In order to understand how termites are guided to choose the most economical path without the use of pheromones, we artificially excavated two tunnels of varying widths (W₁ and W₂; 2, 3, or 4 mm) that intersected at a 90° angle in each of nine soil‐filled arenas. We observed the directional selection of termites at intersections in arenas with combinations of W₁ (mm) and W₂ (mm) (W₁, W₂). For (W₁, W₂) = (2, 2) and (4, 4), termites statistically equally chose the three directions left, right, and straight, while for the (3, 3) arena combination, termites preferentially decided to go straight. On the other hand, for (W₁, W₂) = (2, 3) (2, 4), and (3, 4), termites advancing from narrower tunnels into intersections tended to turn right or left, while termites coming from the wider tunnel were more inclined to go straight. On the assumption that a wider tunnel is more frequently used by termites in the field, we can deduce that the wider tunnel may represent the most efficient/shortest path. Thus, this simple selection mechanism can prevent termites from deviating from the most economical path.     

Evaluation of hexaflumuron for controlling the subterranean termite Coptotermes curvignathus (Isoptera: Rhinotermitidae) in Malaysia

Cellulosic bait matrices containing 0.5% hexaflumuron were tested against field colonies of the subterranean termite Coptotermes curvignathus Holmgren in Malaysia. Foraging activities of the termites were first monitored for several months by using survey stakes and wood bundles placed in underground monitoring traps. Infested stakes were replaced with bait devices. Termite workers to act as recruits were collected from infested stakes and placed in the bait devices. Subsequent activities of the termites were monitored by examining and weighing the wood in the monitoring traps. Data show that the four colonies had populations of 166,288-709,052 foragers and mean wood consumption rates of 305.9-508.2 g/mo per trap. These colonies ceased their activities at all monitoring devices 25-44 d after baiting had commenced. Colonies consumed approximately 27-79 g of bait matrix or 137.5-395 mg of hexaflumuron. Thus, C. curvignathus colonies could be monitored, characterized, baited, and foraging eliminated by using a bait matrix containing hexaflumuron.     

Influence of soil compaction on tunnel network construction by the eastern subterranean termite (Isoptera: Rhinotermitidae)

Eastern subterranean termites, Reticulitermes flavipes (Kollar), workers were introduced into arenas containing low, moderate, and high compaction builder's sand (1.05 g/cm3, 1.18 g/cm3 or 1.35 g/cm3 bulk densities, respectively), and they immediately began tunneling. Termites built the tunnel network significantly fastest in soil of low compaction compared with moderately or highly compacted soil. In soil of low compaction, 221.67 +/- 4.73 cm of total tunnel distance was constructed in 1 d compared with only 96 cm of tunneling in highly compacted soil. At 14 d, total tunnel distance averaged 216.83 +/- 4.56 cm in soil of low compaction compared with 169.70 +/- 4.10 and 181.18 +/- 6.13 cm in moderately and highly compacted soil, respectively. Decreases in total tunnel distance between 1 and 14 d were caused by backfilling of seldom-used tunnels. Termites did the majority of tunneling during the first day of introduction into arenas. In soil of low and moderate compaction, termites essentially constructed the entire tunnel network within the first day, only modifying it by backfilling or maintaining tunnels. In highly compacted soil, 53% of the final tunnel network was constructed during the first day, 87% was constructed by the third day, and 97% was constructed by the seventh day. Soil compaction did not affect the number of primary tunnels or the number and diameter of secondary tunnels. The angle between the secondary tunnel and primary tunnel also was not significantly affected by soil compaction. However, the number of secondary tunnels in soil of low compaction (5.89 +/- 0.51) was significantly greater than in moderately (2.74 +/- 0.36) and highly (3.58 +/- 0.59) compacted soils.     

Distribution of Formosan subterranean termite (Isoptera: Rhinotermitidae) in Mississippi

An extensive monitoring and survey program in Mississippi was conducted from 2000 to 2004 to investigate the distribution of the Formosan subterranean termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae). Seventy-two towns from 22 counties in southern Mississippi were monitored with a total of 3914 traps that catch alates during the swarming season. In addition, 259 licensed pest management professionals in Mississippi were surveyed to determine the locations of termite infestations treated. The alates of C. formosanus were recovered in 12 counties with light traps, and termite infestations were documented in an additional 13 from data collected in the termite survey. Infestations of C. formosanus have been documented in urban, urban cluster, rural, and forested areas of Mississippi. However, the distribution in mean total capture of alates for 4 yr differed significantly among the four ecological areas with the highest percentage in forested areas (31%) and the lowest percentage in urban cluster areas (17%). Most of the infestations of C. formosanus were geographically distributed along the coastal areas of southern Mississippi from Gulfport to Pascagoula. The greatest total number of alates captured in light traps was documented in Pearl River County. Mass swarming of C. formosanus occurred primarily in May or June, depending on weather conditions. The number of documented counties with the evidence of large and widely dispersed swarms of C. formosanus in different ecological areas, and the increase in total annual alate captures from 2000 to 2003, suggest that this invasive termite species is now firmly established in Mississippi.     

Factors affecting post-flight behavior in primary reproductives of the Formosan subterranean termite, Coptotermes formosanus (Isoptera: Rhinotermitidae)

After swarming, reproductive dealates of the Formosan subterranean termite, Coptotermes formosanus, run together in tandem. The tandem running is an age-related behavioral activity in C. formosanus. This behavior was not evoked in pre-flight alates less than 6-d-old that were artificially dealated. Female age was more important than male age for evoking tandem behavior. Females and males, older than 35 days, did not exhibit this behavior. Mating status was not important for female and male dealates to form the tandem pairs. The titers of the major tergal gland component, trilinolein, did not decline significantly and remained high, not only in virgin females, but also in mated females for a period of time after swarming. On the other hand, increasing amounts of the female-specific compound trilinolein were detected in male dealates 7, 14, and 42 days after pairing. This suggests that trilinolein in the females might be transferred to the males as a nuptial gift in C. formosanus. In addition, females have structurally different lateral setae that may constitute a morphological factor involved in the tandem behavior in this species. Covering the setae with dimethyl sulfoxide prevented the tandem behavior.     

Effects of antibiotics on hydrogen production and gut symbionts in the Formosan subterranean termite Coptotermes formosanus (Isoptera: Rhinotermitidae)

Abstract Symbiotic microorganisms that inhabit the gut of Coptotermes formosanus enable this termite to degrade lignocelluloses and further produce hydrogen as an important intermediate to be recycled in its hindgut or as a byproduct to be emitted to the atmosphere. Both symbiotic protists and prokaryotes in the guts of termites demonstrated some different roles with respect to hydrogen production. In this study, the effects of two antibiotics, ampicillin and tetracycline, on hydrogen emission and the gut symbionts of C. formosanus were investigated. Hydrogen emission from termite guts was significantly enhanced when termites fed on wood diets treated with either ampicillin or tetracycline. The greatest H₂ emission rates, 2 519 ± 74 and 2 080 ± 377 nmol/h/g body weight, were recorded with the treatments of ampicillin and tetracycline, respectively, which showed 6–7 times more H₂ production than that of controls. Antibiotic‐treated diets negatively affected the prokaryotic communities and reduced their abundances, particularly on those ectosymbionts inhabiting the gut walls or in the gut fluid of C. formosanus, such as spirochetes. However, no significant reductions in the counts of gut cellulolytic protists, Pseudotrichonympha grassii and Holomastigotoids hartmanni, were recorded; and with a further observation by confocal laser scanning microscopy, the endosymbionts inhabiting P. grassii generally survived the antibiotic treatments. These results suggest that some prokaryotes may serve as the main hydrogen consumers, while P. grassii, together with its endosymbionts, may function as the main contributors for hydrogen production in the hindgut of C. formosanus.     

Effect of fipronil on subterranean termite colonies (Isoptera: Rhinotermitidae) in the field

To assess possible colony-level effects of fipronil, a commonly used nonrepellent termiticide, we conducted a field study of eight houses in the Raleigh, NC, area with infestations of the eastern subterranean termite Reticulitermes flavipes (Kollar) (Isoptera: Rhinotermitidae). We installed an extensive grid of in-ground monitoring stations on each property (mean = 68.6 monitors per property) and collected samples from these stations as well as from mud tubes in the structure and wood debris in the yard for approximately 6 mo. We genotyped all samples by using microsatellite markers to identify the number and locations of colonies present on each property. Houses were treated with either a full treatment (n = 5) or exterior/localized interior treatment (n = 3). After treatment, the monitors were checked monthly for 3 mo and then quarterly for 3 yr to track the fate of colonies. Wood debris in natural areas was checked semiannually for 3 yr. All 11 of the treated colonies (those attacking structures) disappeared within 90 d of treatment and were not found again. These colonies were presumed to be eliminated. In contrast, 60% of untreated colonies (those located > 6 m from the foundation wall at the time of treatment) continued to persist throughout the study, as did 25% of the likely treated colonies (those occupying monitors 0.5 m from the foundation wall where the treatment was applied). Our results provide strong evidence for potent colony wide effects of fipronil on subterranean termites leading to colony suppression and likely colony elimination under field conditions.     

Effect of orange oil extract on the formosan subterranean termite (Isoptera: Rhinotermitidae)

The Formosan subterranean termite, Coptotermesformosanus Shiraki (Isoptera: Rhinotermitidae), accidentally brought into the United States, has become a major urban pest, causing damage to structures and live trees. Because of increasing restrictions on the use of conventional termiticides, attention is focused on finding safer alternative methods for termite management. Oil from citrus peel, referred to here as orange oil extract (OOE), contains -92% d-limonene, and it is generally known to be toxic to insects. In laboratory experiments, 96 and 68% termites were killed in 5 d when OOE at 5 ppm (vol:vol) was dispensed from the top or bottom, respectively, with termites held at the opposite end of a tight-fitting plastic container. Apart from high mortality, workers exposed to vapor consumed significantly less filter paper than controls. However, when termites were exposed to OOE vapor, even at 10 ppm, in the void of a model wall, there was very little mortality. Termites did not tunnel through glass tubes filled with sand treated with 0.2 or 0.4% OOE. Sand treated with OOE was extracted each week for 8 wk to determine the remaining amount of d-limonene. Results indicated that there was a sharp decline in the quantity of d-limonene during the first 3 wk to a residual level that gradually decreased over the remaining period. With a suitable method of application and in combination with other control practices, OOE can be effectively used for the control of subterranean termites.     

Formosan subterranean termite swarming behavior and alate sex-ratio (Isoptera: Rhinotermitidae)

Summary The Formosan subterranean termite (Coptotermes formosanus Shiraki) was first sighted in the cities of New Orleans and Lake Charles, Louisiana, in 1966. Its distribution in the historic French Quarter of New Orleans 23 years later (1989) and in 1991 and 1992 is reported here using counts of alates from light-traps. Light-trappings revealed a Quarter-wide distribution of the termite.C. formosanus shows an extended pause in flight behavior in the middle of the two-month flight season that is predictable and independent of weather. A colony-level mechanism, namely, developmental inhibition of nymphs by alates, may regulate this intermission in flight. The sex-ratio of light-trapped alates varies over time, but is generally female-biased, especially at the height of the flight season.     

Resistance of insecticide-treated foam board insulation against the eastern subterranean termite and the Formosan subterranean termite (Isoptera: Rhinotermitidae)

Three foam board types, one untreated control, one containing 2,000 ppm disodium octaborate tetrahydrate (DOT), and one containing 1,000 ppm deltamethrin, were exposed to field populations of the eastern subterranean termite, Reticulitermes flavipes (Kollar), and the Formosan subterranean termite, Coptotermes formosanus Shiraki. There was no significant difference in termite damage between foam boards treated with 2,000 ppm DOT and the untreated control. Form boards containing 1,000 ppm deltamethrin were not damaged by R. flavipes, whereas only minor damage occurred after exposure to C. formosanus.     

Comparing mark-recapture and constant removal protocols for estimating forager population size of the subterranean termite Coptotermes lacteus (Isoptera: Rhinotermitidae)

Forager population sizes of colonies of Coptotermes lacteus(Froggatt) in New South Wales were estimated using two methods: mark-recapture and constant removal, in two disturbed habitats: a pine plantation and cleared farmland. Mark-recapture population estimates were unrealistic and unreliable: they could be improbably large, over 200 million foragers, and they varied enormously between samples for each colony without any pattern. The constant removal population estimates could also be unrealistic: they could be negative or quite different when calculated using regression and maximum likelihood methods. However, the unrealistic results could be predicted reliably, and explained by the lack of re-contact with the sampling devices (bait stations) - a violation of an assumption of the method. This happened more frequently in the plantation than in the farmland, probably because of the greater abundance of alternative food sources in the plantation. Of the two methods, constant removal provided reasonable forager population estimates, relative to direct counts, at least some of the time, plus a mechanism by which reliability could be tested, whereas mark-recapture provided neither. Further refinement and testing of constant removal methods are urged to provide a more reliable population estimation technique for termites.     

Identification of two subterranean termite species (Isoptera: Rhinotermitidae) using cellulase genes

To identify subterranean termite species, we designed a pair of common primers that amplified 381-bp fragments from cDNAs encoding the endo-beta-1,4-glucanases (EGases) of Coptotermes formosanus Shiraki and Reticulitermes speratus (Kolbe). cDNAs from C. formosanus and R. speratus, and genomic DNA from R. speratus, were amplified by polymerase chain reaction (PCR) by using this primer pair and then cloned and sequenced. Sequences amplified from C.formosanus cDNA displayed 97-99% identity to cDNA encoding the EGase of C. formosanus (CfEG), and 92-94% identity to cDNA encoding the EGase of R. speratus (RsEG). By contrast, cDNA from R. speratus displayed 99-100% identity to RsEG cDNA and 93-94% identity to CfEG cDNA. CfEG and RsEG cDNAs can therefore be used as markers for the identification of these termite species. This is the first report of the successful identification of termite species by using cDNA and genomic DNA sequences of termite origin.     

DNA-based identification of the eastern subterranean termite, Reticulitermes flavipes (Isoptera: Rhinotermitidae)

A DNA-based system for the identification of Reticulitermes flavipes (Koller) was established based on the following criteria: adequate molecular variation, comprehensive specimen sampling, explicit morphological species identification, and cladistic analysis. Termite soldiers were identified by labral shape, and these specimens and associated pseudergates were used in subsequent molecular analyses. Mitochondrial DNA from the AT-rich region was sequenced for 173 R. flavipes, Reticulitermes hageni Banks and Reticulitermes virginicus (Banks) soldiers and pseudergates collected from several widely dispersed Texan localities. Cladistic analysis was performed after exploration of an optimal sequence alignment inclusive of DNA sequences from Texas and published sequences from Georgia and Canada. Among the Texan individuals, 29, 5, and 1 mitochondrial DNA haplotypes were identified respectively with R. flavipes, R. hageni, and R. virginicus. One haplotype from El Paso likely represented a different Reticulitermes species. Species were monophyletic; however, individual relationships were unresolved in a strict consensus of 700 most parsimonious trees. Texas haplotypes were isolated by distance, and a correlation between genetic and geographic distance was observed among the Texas, Georgia, and Canada haplotypes. These results suggest that the methods outlined in this study will allow for the identification of R. flavipes from localities between Texas and Canada.     

Parthenogenetic reproduction in neotenics of the subterranean termite Reticulitermes speratus (Isoptera: Rhinotermitidae)

Experiments with artificial colonies consisting of only female workers and those of female nymphs and workers of a subterranean termite, Reticulitermes speratus, showed that neotenics (ergatoids and nymphoids) can reproduce parthenogenetically. The larvae hatched from the eggs produced partheno‐genetically by the neotenics were all female and did not have any aberrant external morphology. They had 42 chromosomes and were diploid. The analysis of genotypes at microsatellite loci revealed that the eggs produced parthenogenetically had only one of the two alleles that the mother neotenics possessed, indicating that the eggs were produced by automixis.     

Factors affecting mate choice in the subterranean termite Reticulitermes chinensis (Isoptera: Rhinotermitidae)

Termites are a group of eusocial insects. Mate choice is the most important step which affects the subsequent new colony foundation and development in termites. This study investigated the effects of the four factors on mate choice in the subterranean termite Reticulitermes chinensis Snyder, including colony origin, entry time, physical damage of antennae, and fresh body weights of male dealates. The results showed that the pairing number of dealates from the different colonies was significantly higher than that from the same colonies. The male dealates with early entry time achieved a significantly higher pairing number than the male dealates with late entry time. The male dealates with complete antennae achieved a the significantly higher pairing number than the male dealates lacking the six terminal segments of antennae. Moreover, the heavier male dealates achieved a significantly higher pairing number than the lighter male dealates. However, there were no significant differences in the pairing time for all the experimental treatments. These results suggested that the male dealates, which are heterogenous, healthy, heavy, and more familiar with the environment in R. chinensis, have the obvious advantages in the mate choice. This rule of mate choice is helpful for termites to avoid inbreeding and to maintain the genetic diversity of offspring, which is very important for the environmental adaptability and development of termite colonies.     

Hydrogen emission by three wood-feeding subterranean termite species (Isoptera: Rhinotermitidae): Production and characteristics

Abstract Hydrogen emission by wood-feeding termites, Coptotermes formosanus, Reticulitermes flavipes and Reticulitermes virginicus, was investigated upon a cellulosic substrate as their food source. The emission rates among the three species tested were significantly different and R. virginicus demonstrated the greatest H₂ emission at 4.78 ± 0.15 μmol/h/g body weight. In a sealed test apparatus, H₂ emission for each termite species showed a quick increase at the initial incubation hours (3–6 h), followed by a slower growth, possibly due to the feedback inhibition by gas accumulation. Further investigation revealed that continuous H₂ emission could be maintained by reducing the H₂ partial pressure in the sealed container. The bioconversion of cellulose to molecular H₂ by the subterranean termites tested could reach as high as 3 858 ± 294 μmol/g cellulose, suggesting that the termite gut system is unique and efficient in H₂ conversion from cellulosic substrate.