The relationship among body size (as indicated by wing length), age (as indicated by parity dissections), and malaria infection were observed in host-seeking Anopheles maculatus Theobald females collected in aboriginal villages of peninsular Malaysia. Both ELISA and salivary gland dissections were used to determine malaria infection. The wings of parous females were significantly longer than those of nulliparous females, suggesting that larger females live longer than smaller ones, and thus have a higher vectorial capacity. Body size differences were not detected between infected parous and uninfected parous females. Females infected with only oocysts were significantly larger than females infected with sporozoites. No correlation was found between the number of oocysts or sporozoites and body size in this small sample.
A dot-immunobinding assay (DIBA) was compared with a direct fluorescent antibody technique (DFAT) for the detection of Rickettsia tsutsugamushi infection in Leptotrombidium fletcheri (Womersley & Heaslip). Laboratory colonies of infected and noninfected chiggers were examined. The relative proportions of positive, negative, and indeterminate results were significantly different between DIBA and DFAT for infected but not for noninfected chiggers. DIBA was more sensitive and had a better negative predictive value and a lower false negative percentage than DFAT. It was concluded that DIBA is a suitable alternative to DFAT for detecting scrub typhus infection in chiggers.
Anopheles maculatus Theobald sensu lato is a species complex now consisting of eight sibling species; An. maculatus is still represented by two cytologically distinct forms; i.e., the widely distributed sensu strictu or B, and E from southern Thailand and adjacent areas in northern Malaysia. Cuticular lipid profiles in conjunction with principal component analysis was used to separate An. maculatus form E from sensu stricto form B in a preliminary survey of the An. maculatus complex at five locations spanning peninsular Malaysia. The relative rank orders, from the areas of the five gas chromatographic peaks used to determine lipid differences for specimens from peninsular Malaysia, matched well with those from cytogenetically identified colony specimens of An. maculatus forms B and E. The two-dimensional principal component pattern of specimens identified as form E was highly clumped, which indicated that very similar cuticular lipids were present within this putative malaria vector. Both forms coexisted in peninsular Malaysia, but form E may be dominant except in the south.
Aedes albopictus (Skuse) is an Asian mosquito that recently has colonized North America via used tire transport. Temperate Ae. albopictus populations overwinter as diapausing eggs, but tropical populations cannot diapause. Eggs of tropical (SABAH) and temperate (INDY) Ae. albopictus were obtained in diapause-inducing conditions and placed inside a scrap tire to monitor overwintering survival in northern Indiana during the winters of 1989-1990 and 1990-1991. Diapause eggs of Ae. triseriatus (Say), a native North American mosquito, were included for comparison. Tropical Ae. albopictus from Malaysia did not survive either winter. Temperate Ae. albopictus from Indianapolis, IN, did not survive the winter of 1989-1990, but 78% survived the winter of 1990-1991. In contrast, 92 and 96% of Ae. triseriatus survived the winters of 1989-1990 and 1990-1991, respectively. Neither mean temperature nor absolute minimum temperature (a winter's lowest temperature) accurately predicted Ae. albopictus overwintering survivorship in the field. The possible effect of snow and other insulating materials on the overwintering survivorship of Ae. albopictus eggs is discussed.
A 2-yr study of Japanese encephalitis (JE) virus in Sepang District, Selangor, Malaysia, was carried out to identify the mosquito vectors and to determine their seasonal abundance, parity, and infection rates. In total, 81,889 mosquitoes belonging to 9 genera and > 50 species were identified from CDC trap collections augmented with dry ice during 1992 and 1993. Culex tritaeniorhynchus Giles and Culex gelidus Giles were the most abundant species, and both increased in numbers with increases in rainfall. Overall, 45 JE virus isolations were made from 7 species-Cx. tritaeniorhynchus (24), Cx. gelidus (12), Culex fuscocephala Theobald (2), Aedes butleri Theobald (4), Culex quinquefasciatus Say (1), Aedes lineatopennis Ludlow (1), and Aedes (Cancraedes) sp. (1). Based on elevated abundance and JE infection rates, Cx. tritaeniorhynchus appears to be the most important vector of JE virus in Sepang.
Factors affecting the mechanical transmission of rotavirus by the legs and wings of the housefly, Musca domestica L., were examined in a laboratory study. Rotavirus was picked up when houseflies walked on thin smears of clarified rotavirus suspensions. The addition of glycerol, which increased viscosity of the virus suspension, and particulate human feces slightly increased the proportion of flies contaminated with virus. However, the addition of glycerol greatly reduced the average number of virus particles picked up per fly, whereas feces greatly increased the number of particles. The proportion of flies with virus-contaminated legs, which transferred virus to > 1 contact surface, was increased by longer contact time with the surface and when the contact surface was agar instead of glass. Most virus particles were deposited on 1st contact with the surface. Most flies dislodged virus particles inoculated on the underside of their wings soon after the start of simulated flight. Our data indicated that the nature of the virus-suspending medium has a greater effect on the level of virus contamination than on the ability to become contaminated. The importance of walking as a mode of virus transport depends on the nature of the contact surface, the risk of the contaminated fly settling first on a surface likely to come into contact with humans, and fly numbers.
Maxillae and mandibles of males of 44 species of 12 mosquito genera and females of three autogenous genera and two partially autogenous species were examined under light microscopy and scanning electron microscopy. The maxillae and mandibles of male mosquitoes are delicate, tape-like structures with lengths characterizing genera or higher level classification units. Five patterns are recognized: (A) long maxillae and mandibles with mandibles longer than maxillae in Anopheles; (B) long maxillae and mandibles with maxillae longer than mandibles in Toxorhynchites; (C) short or intermediate lengths of maxillae with short mandibles in Aedes, Armigeres, Culex, Ochlerotatus, Orthopodomyia, and Uranotaenia; (D) short or intermediate length of maxillae with no mandibles in Mimomyia and Tripteroides; and (E) no maxillae and mandibles in Malaya and Topomyia. Maxillary and mandibular lengths of male mosquitoes show a positive correlation. Length of maxillae and mandibles of autogenous females are reduced to the same level as conspecific males. In contrast, females of partially autogenous species have complete maxillae and mandibles as in females of anautogenous species.
Within mosquitoes, arboviruses encounter barriers to infection and dissemination that are critical determinants of vector competence. The molecular mechanisms responsible for these barriers have yet to be elucidated. The prototype Sindbis (SIN) strain, AR339, and viruses derived from this strain, such as TR339 virus, have limited infection and transmission potential in the medically important arthropod vector, Aedes aegypti (L.). However, the Malaysian SIN virus strain, MRE16, disseminates in nearly 100% of Ae. aegypti 14 d after oral infection. Here, we compare the spatial and temporal infection patterns of MRE16 and TR339 viruses in Ae. aegypti. The results indicate that a midgut escape barrier is primarily responsible for the significantly lower dissemination and transmission potentials observed after oral infection with TR339 virus. MRE16 and TR339 viruses now represent a well-characterized model system for the further study of virus determinants of vector infection, particularly determinants affecting the midgut escape barrier in Ae. aegypti.
Anopheles sundaicus s.l. is a principal malaria vector taxon on islands and along the coastal areas of Southeast Asia. It has a wide geographical distribution and exhibits a high level of ecological and behavioral variability. Study of this taxon is crucial for understanding its biology and implementing effectise vector control measures. We compared populations of An. sundaicus from Vietnam, Thailand, and Malaysian Borneo by using two mitochondrial DNA markers: cytochrome oxidase I and cytochrome b. Genetic divergence, geographic separation, and cladistic analysis of relationships revealed the presence of two cryptic species: Anopheles sundaicus s.s. on Malaysian Borneo and An. sundaicus species A in coastal areas of Thailand and Vietnam. A polymerase chain reaction (PCR) assay was developed to easily identify these two species throughout their geographic distributions. The assay was based on sequence characterized amplified region derived from random amplified polymorphic DNA. This PCR identification method needs to be validated and adapted for the recognition of other possible species in the Sundaicus Complex.
To advance our limited knowledge of global mosquito biogeography, we analyzed country occurrence records from the Systematic Catalog of the Culicidae (http://www.mosquitocatalog. org/main.asp), and we present world maps of species richness and endemism. A latitudinal biodiversity gradient was observed, with species richness increasing toward the equator. A linear log-log species (y)-area (x) relationship (SAR) was found that we used to compare observed and expected species densities for each country. Brazil, Indonesia, Malaysia, and Thailand had the highest numbers of species, and Brazil also had the highest taxonomic output and number of type locations. Brazil, Australia, the Philippines, and Indonesia had the highest numbers of endemic species, but excluding small island countries, Panama, French Guiana, Malaysia, and Costa Rica had the highest densities of total species and endemic species. Globally, 50% of mosquito species are endemic. Island countries had higher total number of species and higher number of endemic species than mainland countries of similar size, but the slope of the SAR was similar for island and mainland countries. Islands also had higher numbers of publications and type locations, possibly due to greater sampling effort and/or species endemism on islands. The taxonomic output was lowest for some countries in Africa and the Middle East. A consideration of country estimates of past sampling effort and species richness and endemism is proposed to guide mosquito biodiversity surveys. For species groups, we show that the number of species of Anopheles subgenus Anopheles varies with those of subgenus Cellia in a consistent manner between countries depending on the region. This pattern is discussed in relation to hypotheses about the historical biogeography and ecology of this medically important genus. Spatial analysis of country species records offers new insight into global patterns of mosquito biodiversity and survey history.
This study examined the effects of different life stages (first, second, third, fourth, and five instars; adult females and adult males) and feeding regimes (starved and blood fed) on the active movement activity of the tropical bed bug, Cimex hemipterus (F.), under mixed-stage conditions. We used an extended arena made from Tygon tube coils and observed the movement frequency and movement distance at selected time intervals up to 120 h. The fifth instars and adult males and females showed significantly (P < 0.01) greater movement frequency compared with the other stages. The first and second instars showed limited movement (< 8 m) over the experimental period. Starved bed bugs showed greater movement frequency compared with blood-fed bed bugs, with the exception of adult females. Blood-fed adult females exhibited significantly (P < 0.01) greater movement frequency and distance compared with starved females. Blood-fed females moved up to 42.3 m over 120 h. Regression analysis between movement distance of the fifth instars and adults and the time intervals revealed a positive relationship (r2 = 0.6583; P < 0.01), suggesting that delays in bed bug control efforts will increase the risk of the greater infestation. During bed bug inspection, the presence of only late instars and adults in premises would indicate a new infestation, whereas an established infestation likely would consist of mixed stages.
The effect of temperature and humidity on the survival and water loss of the tropical bed bug, Cimex hemipterus (F.), was studied using two field-collected strains. Insects were exposed to temperatures ranging from 20 to 45 degrees C and relative humidities (RHs) of 33, 75, and 100%. C. hemipterus survived longest under the interaction of low temperature (20 degrees C) and high RH (75-100%). Survival and water loss were significantly affected (P < 0.01) by temperature and RH (either singly, or in interaction). Strain and sex significantly (P < 0.01) influenced bed bug survival, but not on water loss. Eggs, first instars, and adults reached their upper thermal lethal limit within 1 h at 39 degrees C, 44 degrees C, and 46 degrees C, respectively. The survival and water loss profiles showed that starved C. hemipterus started to die after losing 35-45% of their body weights.
Behavioral responses of Aedes aegypti male populations developed for Release of Insects Carrying a Dominant Lethal (RIDL) technology and a Malaysian wild-type population of two age groups (4-5 and 8-10 d old) were tested under laboratory conditions against chemical irritants and repellents using the high-throughput screening system device. Results indicate that all male Ae. aegypti test populations showed significant (P < 0.01) behavioral escape responses when exposed to alphacypermethrin, DDT, and deltamethrin at the test dose of 25 nmol/cm2. In addition, all populations showed significant (P < 0.05) spatial repellent responses to DDT, whereas alphacypermethrin and deltamethrin elicited no directional movement in the assay. These data suggest that genetic modification has not suppressed expected irritancy and repellency behavior. Age effects were minimal in both contact irritant and spatial repellent assays. The magnitude of irritant response, based on percentage responding, was stronger in the RIDL test cohorts as compared with the wild-type Malaysian population, but the impact, if any, that this increased behavioral sensitivity might have on the success of a RIDL strategy has yet to be defined. Information of the type reported in the current study is vital in defining the effects of genetic modification on vector behavior and understanding how these behaviors may influence the success of RIDL technology as they relate to other vector control interventions implemented in the same disease-endemic locale.
The population size, age-class structure, and movement of the American cockroach, Periplaneta americana (L.) (Dictyoptera: Blattidae), were studied in three sewers in Penang, Malaysia, from September 2008 to October 2009. Eighteen to 20 glass-jar traps (two per manhole) were deployed for a 24-h period during each sampling occasion at each sewer. Adults and nymphs were active throughout the study period, with an average monthly trap catch of 57-97 adults and 79-99 nymphs. The mean proportions of adults and nymphs at the three sewers ranged from 0.47 to 0.57. Of the 2177 male and 2717 female cockroaches marked and released over the three sewers, recapture rates were 29.4-45.8 and 30.8-47.0%, respectively. The proportion of marked males and females did not differ significantly from the proportion of recaptured marked males and females. However, the mean number of times a marked female was recaptured was significantly greater than that of males. Of the 783 males and 1,030 females that were marked and recaptured, 19.4 and 24.7%, respectively, had moved between manholes, and significantly more females than males moved between manholes. Of the 406 recaptured marked adults that moved between manholes, 90.4% moved a distance of 2-20 m from their initial release site; one male moved 192 m, the longest distance recorded. Trap catch on each sampling occasion was positively correlated with daily mean temperature. The number of cockroach movements between manholes also was correlated with the mean daily minimum temperature.
There is accumulating evidence that criminals wrap dead bodies in an attempt to conceal evidence. To anticipate the forensic implications of this phenomenon, we examined whether flies that are naturally associated with cadavers exhibit a delay in attendance or differ in species composition and abundance patterns because of the presence of wrapping material. Wrapped and exposed carcasses of dead monkeys placed in an oil plantation in Kedah, Malaysia, were visited over 50 d. On daily visits to each of the six carcasses, visiting adult flies were sampled using hand nets. Flies of 12 families were encountered. Calliphoridae (Chrysomya rufifacies Macquart and C. megacephala (F.) was the most prevalent family, followed by Sphaeroceridae. Some families tended to be more abundant in WRCs (i.e., Calliphoridae, Muscidae, and Phoridae), whereas others (i.e., Piophilidae, Sepsidae, and Psychodidae) were more prevalent in exposed carcasses. Wrapping delayed the arrival of all fly species encountered, with delays varying from 1 to 13 d depending on species. Wrapping did not affect species composition of flies, but prolong the occurrence of some species. The results of the current study emphasize the need to take into consideration the presence of a wrap when estimating postmortem interval.
The life history characteristics of the rove beetle Paederus fuscipes Curtis were studied under laboratory conditions using three field strains from Malaysia: Desa Wawasan (DW), Sri Pinang (SP), and Ampang Jajar (AJ). The total development time of immature stages differed significantly among the three strains, especially between DW (17.43 +/- 0.16 d), SP (18.60 +/- 0.19 d), and AJ (18.68 +/- 0.22 d). Adult females and males from DW also exhibited a shorter life span, although the difference among strains was not significant. In terms of fecundity, the numbers of eggs laid per female for DW, SP, and AJ were 121.28 +/- 15.98, 127.30 +/- 18.01, and 147.45 +/- 17.12, respectively. Additionally, because of the shorter life span in DW strain, two apparent peaks in age-stage specific fecundity were detected. The beetles compensated for their shorter life span by increasing their reproductive activity to sustain the progeny in the population. The intrinsic rates of increase (r) of P. fuscipes from DW, SP, and AJ were 0.0773 +/- 0.0046 d(-1), 0.0788 +/- 0.0051 d(-1), and 0.0873 +/- 0.0054 d(-1), respectively; and the net reproduction rates (R0) were 40.09 +/- 7.39 offspring, 45.29 +/- 8.74 offspring, and 42.34 +/- 8.25 offspring, respectively. The mean generation time of P. fuscipes from AJ was 43.08 +/- 1.07 d, which was significantly higher than that from DW (47.95 +/- 1.36 d) and SP (48.57 +/- 1.43 d). The total immature development time of P. fuscipes in this study was shorter than values reported in previous studies.
Two new species of black flies, Simulium (Comphostilbia) terengganuense sp. nov. and Simulium (Gomphostilbia) aziruni sp. nov. (Diptera: Simuliidae), are described on the basis of reared adult, pupal, and larval specimens collected from Peninsular Malaysia. Both species are placed in the batoense species-group within the subgenus Gomphostilbia, one of two dominant subgenera of the genus Simulium in Peninsular Malaysia as well as in the Oriental Region. Strikingly, three morphological characteristics that rarely occur in the subgenus Gomphostilbia are found in these two new species: the very narrow female frons and the mushroom-like pupal terminal hooks in S. (G.) terengganuense sp. nov. and the pupal gill composed of an inflated horn-like structure and eight slender filaments in S. (G.) aziruni sp. nov.
Simulium (Simulium) kisapense sp. nov. is described on the basis of reared adult, pupal, and larval specimens collected from Langkawi Island, Peninsular Malaysia, and placed in the multi-striatum species-group in the subgenus Simulium. This new species is characterized by the bare basal section of the female radial vein, the male ventral plate with setae, the eight pupal gill filaments divergent at an angle of >90 degrees, and the shoe-shaped cocoon with small lateral window(s). Taxonomic notes are given to separate this new species from S. (S.) hirtinervis Edwards and S. (S.) malayense Takaoka and Davies, both from Peninsular Malaysia, and several other known species from Bhutan, India, Nepal, and Thailand.
A nationwide investigation was carried out to determine the current susceptibility status of Culex quinquefasciatus Say populations against four active ingredients representing four major insecticide classes: DDT, propoxur, malathion, and permethrin. Across 14 study sites, both larval and adult bioassays exhibited dissimilar trends in susceptibility. A correlation between propoxur and malathion resistance and between propoxur and permethrin resistance in larval bioassays was found. The results obtained from this study provide baseline information for vector control programs conducted by local authorities. The susceptibility status of this mosquito should be monitored from time to time to ensure the effectiveness of current vector control operations in Malaysia.
A comprehensive 8-yr survey of acarine ectoparasites (ticks and mites) of bats was carried out in 18 localities from 2002 to 2009. Most of the surveys were conducted during 14 national biodiversity scientific expeditions throughout Malaysia. The objective was to identify acarines of known public health importance from bats and thus determine whether there is any potential public health risk in Malaysia. Trapping of bats was conducted using Harp traps and Mist nets. In total, 1,579 individuals comprising of 6 families and 52 species of bats were examined alive. In general, 25.6% of the bats were infested with acarines. Infestation rates of ticks, mesostigmatid mites, and chiggers on bats examined were 0.4, 10.4, and 14.7%, respectively. Their prevalence and mean intensity were tabulated. Genera of ticks extracted were Amblyomma, Dermacentor, Ixodes, and Ornithodoros. Of these genera, only two species can be identified to species level and they are Amblyomma cordiferum and Ixodes simplex. In total, 8 genera and 15 species of mesostigmatid mites were found; the species were Ancystropus eonycteris, Ancystropus zeleborii, Echinonysus nasutus, Laelaps aingworthae, Laelaps nuttalli, Laelaps sanguisugus, Laelaps sculpturatus, Longolaelaps longulus, Longolaelaps whartonii, Meristaspis lateralis, Meristaspis macroglossi, Paraperiglischrus rhinolophinus, Spinturnix acuminatus, Spinturnix americanus, and Spinturnix bakeri. Chiggers on bats were represented by 12 genera and 6 species; the species identified were Gahrliepia fletcheri, Riedlinia lipoxena, Trombigastia cadei, Walchiella impar, Walchiella oudemansi, and Whartonia caobangensis. The study produced an up-to-date list of acarine ectoparasites of bats in Malaysia where a total of 38 genera and 47 species of acarines were listed. Findings of the study demonstrated that 5 genera and 1 species of acarines that may pose potential health risks, can be found on bats.