Rapid insecticide resistance bioassays for three major urban insects in Taiwan.

BACKGROUND: Taiwan's warm and humid climate and dense population provide a suitable environment for the breeding of pests. The three major urban insects in Taiwan are house flies, cockroaches, and mosquitoes. In cases where a disease outbreak or high pest density necessitates chemical control, selecting the most effective insecticide is crucial. The resistance of pests to the selected environmental insecticide must be rapidly assessed to achieve effective chemical control and reduce environmental pollution. METHODS: In this study, we evaluated the resistance of various pests, namely, house flies (Musca domestica L.), cockroaches (Blattella germanica L. and Periplaneta americana), and mosquitoes (Aedes aegypti and Ae. albopictus) against 10 commonly used insecticides. Rapid insecticide resistance bioassays were performed using discriminating doses or concentrations of the active ingredients of insecticides. RESULTS: Five field strains of M. domestica (L.) are resistant to all 10 commonly used insecticides and exhibit cross- and multiple resistance to four types of pyrethroids and three types of organophosphates, propoxur, fipronil, and imidacloprid. None of the five field strains of P. americana are resistant to any of the tested insecticides, and only one strain of B. germanica (L.) is resistant to permethrin. One strain of Ae. albopictus is resistant to pirimiphos-methyl, whereas five strains of Ae. aegypti exhibit multiple resistance to pyrethroids, organophosphates, and other insecticides. CONCLUSIONS: In the event of a disease outbreak or high pest density, rapid insecticide resistance bioassays may be performed using discriminating doses or concentrations to achieve precise and effective chemical control, reduce environmental pollution, and increase control efficacy.

Effectiveness and acceptance of total release insecticidal aerosol cans as a control measure in reducing dengue vectors.

The effectiveness of regular application of insecticidal fogging in reducing dengue is questionable, since delays occur between peak time of outbreak and insecticide administrations. Moreover, many residents do not accept indoor application because of concern about insecticide contamination of household items. The study described in this article was designed to evaluate the effectiveness and acceptance of insecticidal aerosol cans to reduce dengue vectors inside and outside of homes. Residents in Kaohsiung City of South Taiwan were provided with two formulations of aerosol cans (permethrin 3.75% weight/weight [w/w] and cypermethrin 1.716% w/w) and were requested to use these aerosol cans. Although the indoor ovitrap index of the permethrin group returned to the original level in week 3, the index of the cypermethrin group decreased 60% to 20%. The residents accepted the insecticidal aerosol cans but complained of unfavorable effects caused by traditional insecticidal fogging. Results indicate that the insecticidal aerosol cans may serve as a supplementary household control measure for dengue vectors during the time period between the peak of outbreak and the administration of government-organized insecticide fogging.

Immunologic characterization and allergenicity of recombinant Tyr p 3 allergen from the storage mite Tyrophagus putrescentiae.

BACKGROUND: The involvement of the storage mite Tyrophagus putrescentiae in allergies has been increasingly reported in many countries. Molecular analysis has shown that group 3 mite allergens are homologous to trypsin. Similar allergens have not been identified in T. putrescentiae. Our aims were to characterize group 3 allergens in T. putrescentiae and to investigate their significance in allergenicity. METHODS: cDNAs of PreTyr p 3 and rTyr p 3 from T. putrescentiae were cloned and expressed in Escherichia coli. Native Tyr p 3 (nTyr p 3) was purified from spent growth medium with an affinity column coupling of antibody. Biological activities of rTyr p 3 were compared with nTyr p 3 in terms of IgE activity, enzymatic activity and histamine release. RESULTS: Full-length cDNA of PreTyr p 3 encodes a 285-amino acid trypsin-like protease and acquires enzymatic activity after removing the pre- and pro-sequences. rTyr p 3 is a 26-kDa protein with equivalent IgE reactivity but weaker enzymatic activity than that of nTyr p 3. A limited level of cross-reactivity has been found between rTyr p 3, Der p 3 and Blomia. Eight of 10 T. putrescentiae-sensitized individuals showed >50% histamine release after triggering with rTyr p 3. CONCLUSIONS: Our studies demonstrate that Tyr p 3 is a frequent allergen (58%) in T. putrescentiae-sensitized patients. Since rTyr p 3 displays equivalent biological activities as nTyr p 3, the role of group 3 allergens can be studied using rTyr p 3 to elucidate the pathogenic effects and diagnostic applications of Tyr p 3.

A novel amino acid substitution in a voltage-gated sodium channel is associated with knockdown resistance to permethrin in Aedes aegypti.

To identify pertinent mutations associated with knockdown resistance to permethrin, the entire coding sequence of the voltage-gated sodium channel gene Aa-para was sequenced and analyzed from a Per-R strain with 190-fold resistance to permethrin and two susceptible strains of Aedes aegypti. The longest transcript, a 6441bp open reading frame, encodes 2147 amino acid residues with an estimated molecular mass of 241kDa. A total of 33 exons were found in the Aa-para gene over 293kb of genomic DNA. Three previously unreported optional exons were identified. The first two exons, m and n, were located within the intracellular domain I/II, and the third, f', was found within the II/III linkers. The two mutually exclusive exons, d and l, were the only alternative exons in all the cDNA clones sequenced in this study. The most distinct finding was a novel amino acid substitution mutation, D1794Y, located within the extracellular linker between IVS5 and IVS6, which is concurrent with the known V1023G mutation in Aa-para of the Per-R strain. The high frequency and coexistence of the two mutations in the Per-R strain suggest that they might exert a synergistic effect to provide the knockdown resistance to permethrin. Furthermore, both cDNA and genomic DNA data from the same individual mosquitoes have demonstrated that RNA editing was not involved in amino acid substitutions of the Per-R strain.

Differential survival of Aedes aegypti and Aedes albopictus (Diptera: Culicidae) larvae exposed to low temperatures in Taiwan.

Aedes aegypti (L.) and Aedes albopictus (Skuse) differ in their distribution in Taiwan. The former species is distributed in the south of Taiwan, whereas the latter is found throughout the island. One possible explanation proposes that low temperatures in the winter prevent the expansion of Ae. aegypti. Hence, the impact of low temperatures on immatures of both species was studied in the laboratory and in the field. Our study showed that, under most conditions, Ae. aegypti were more sensitive to low temperatures than Ae. albopictus both in the laboratory and in the field. The survival rates at 10 degrees C for the first and fourth instars of Ae. albopictus were significantly better than those of Ae. aegypti. At 2.5 and 5.0 degrees C, the first instars of Ae. albopictus survived better than the same stadium of Ae. aegypti, but the fourth instars of Ae. aegypti survived better. Short exposures to low temperatures did not affect the acclimatization of Ae. aegypti immatures but longer exposures did increase the physiological adaptation to low temperatures. For Ae. albopictus, exposure to low temperatures increases the acclimatization of this species. In field experiments, Ae. aegypti larvae had a significantly higher mortality than Ae. albopictus during exposures to cold fronts in the 2004 winter. We conclude that low temperatures in northern and central Taiwan have a negative impact on the distribution of Ae. aegypti, but this factor alone is not sufficient to prevent this species from occupying the rest of Taiwan.

Seaport as a source of hantavirus: a study on isolated isles.

Since small mammals from seaports have relative higher seroprevalences of hantavirus, this seroepidemiologic study was conducted on the isles in Kinmen and Lienchiang Counties along the coast of southern mainland China to determine whether seaport may play the role as a source of hantavirus. Among six species of small mammals trapped in Kinmen County, only male Mus musculus (6.7%) and Rattus losea (5.3%) were found to be positive. In Lienchiang County, five species of small mammals were trapped and positive findings were obtained only in male R. norvegicus (9.3%), male M. musculus (7.7%), and female R. losea (1.6%). There was no significant difference in the seroprevalence between the two counties (Kinmen 3.8% vs. Lienchiang 3.9%). The positive rate in Liaolo (17.9%) of Kinmen County was significantly higher than the remaining trapping stations and those in Matsu Distillery (10.3%) and Fushing (5.8%) of Lienchiang County were significantly higher than the remaining ones. Moreover, a significant inverse correlation was found between the seropositive rate and the distance of small mammal sampling sites to the seaport (p < 0.01). These findings suggest the role of seaport as a source of hantavirus.

Impact of a short-term community-based cleanliness campaign on the sources of dengue vectors: an entomological and human behavior study.

In June 2002, a considerable number of cases of dengue fever/dengue hemorrhage fever were reported in South Taiwan. To effectively reduce the sources of dengue vectors, the government conducted a weeklong community-based cleanliness campaign. The study reported here was designed to evaluate the effectiveness of the campaign. The sources of dengue vectors were evaluated according to changes in the ovitrap index. This index significantly decreased from 66.7 percent before the campaign to 39.3 percent three months later. Human knowledge and behavior were investigated by a survey questionnaire. Although the respondents had sufficient knowledge about dengue as well as its transmission and prevention, only 57.4 percent knew that the dengue vector breeds in clean water. After the campaign, this percentage significantly increased, to 88.4 percent, and the percentage of those properly handling artificial containers also significantly increased. On the basis of these findings, the authors conclude that short-term community-based cleanliness educational programs may change behavior to eliminate container habitats of the dengue vectors and in turn reduce the ovitrap index. This control strategy is a feasible alternative to rapidly reduce sources of dengue vector in urban areas, especially at the onset of a new epidemic.

The differences of dengue vectors and human behavior between families with and without members having dengue fever/dengue hemorrhagic fever.

Dengue vectors, human knowledge and behavior have been reported to play an important role in the transmission of dengue. This study was designed to understand the differences of dengue vectors and the behavior between families with (target group) and without (control group) members having dengue fever/dengue hemorrhagic fever. Population density of dengue vectors were determined by ovitrap index. The living conditions, knowledge, and behavior related to dengue were investigated by questionnaire survey. Long-term ovitrap indices obtained in the target group was significantly higher than those obtained in the control. Most of the respondents had sufficient knowledge about the transmission and prevention of dengue. However, only low percentages of the families frequently cleaned water-filled containers and ditches around their residence, especially in the target group. These findings indicate that higher indices of dengue vectors and dengue-related behavior are important in the transmission of indigenous dengue.

Insecticide resistance in German cockroaches (Blattella germanica) from hospitals and households in Taiwan.

Since a high percentage of cockroach infestation has been determined in hospitals in Kaohsiung, South Taiwan, insecticide resistances may have developed in the cockroach populations. To determine the resistances in cohorts of German cockroach (Blattella germanica) from hospitals and households to organophosphates, carbamates and pyrethroids, cockroaches were trapped from 30 hospitals and 30 households in this area. Bioassays by topical application were then carried out on the first generation bred in the laboratory. Resistance ratio (RR) was determined by comparing the LD50 of the collected cohorts with that of a susceptible cohort. RR ranges of chlorpyrifos, propoxur and cypermethrin in the hospitals were 2.04 - 28.80, 2.86 - 30.86 and 1.95 - 14.05 and those in the household were 1.92 - 17.72, 6.93 - 62.50 and 2.80 - 27.35 respectively. Significant correlations among the RRs of the three insecticides were separately found in the hospitals and households, indicating the existence of cross resistance among the three groups of insecticides. The patterns of propoxur > chlorpyrifos > cypermethrin in hospitals and propoxur > cypermethrin > chlorpyrifos in households reflect that chlorpyrifos is more frequently used in hospitals than in households. In contrast, cypermethrin is used more frequently in households.

Genetic relationships among populations of Aedes aegypti in Taiwan by using phenotypic and random amplified DNA-polymerase chain reaction markers.

An analysis of gene flow was conducted among collections of Aedes aegypti from 7 localities along the southwestern and southeastern coasts in Taiwan. Markers include 7 types of scaling patterns and 23 random amplified polymorphic DNA (RAPD) loci amplified by the polymerase chain reaction. Differences in scaling pattern and in the frequencies of RAPD markers were detected among populations and cluster analyses revealed 2 main groups on each side of the Central Mountain Range. Regression analysis of geographic distances and pairwise F(ST) values estimated from RAPD markers showed that southwestern populations are isolated by distance and that populations within 15 km are panmictic. This is a shorter distance than detected among collections of Ae. aegypti in similar published studies from Mexico and Argentina.

Voltage-dependent calcium channels in the corpora allata of the adult male loreyi leafworm, Mythimna loreyi.

In the corpora allata (CA) of the adult male loreyi leafworm, Mythimna loreyi, juvenile hormone acid (JHA) biosynthesis and release show a dose dependence on extracellular Ca(2+) concentration. Maxima are obtained with Ca(2+) concentrations of 2-10 mM, and synthesis and release are significantly inhibited under a Ca(2+)-free condition. The Ca(2+)-free inhibition of JHA release can be reversed by returning the glands to medium at 5 mM Ca(2+). The cytosolic free Ca(2+) concentration ([Ca(2+)](i)), which was measured with fura-2, in individual CA cells also shows a dose dependence on extracellular Ca(2+) concentration, with significant [Ca(2+)](i) depression being observed in the absence of extracellular Ca(2+). High K(+) significantly increases the JHA release and causes a transient [Ca(2+)](i) increase within seconds in CA cells. High-K(+)-stimulated JHA release is partially inhibited by the benzothiazepine (BTZ)-, dihydropyridine (DHP)- and phenylalkylamine (PAA)-sensitive L-type voltage-dependent calcium channel (VDCC) antagonists diltiazem, nifedipine and verapamil, respectively; by the N- and P/Q-type VDCC antagonist omega-conotoxin (omega-CgTx) MVIIC; and by the T-type VDCC antagonist amiloride. The N-type antagonist omega-CgTx GVIA is the most potent in inhibiting the high-K(+)-stimulated JHA release. No inhibitory effect is shown by the P-type antagonist omega-agatoxin TK (omega-Aga TK). The high-K(+)-induced transient [Ca(2+)](i) increase is largely inhibited by the L-type antagonists (diltiazem, nifedipine, verapamil), by the N- and P/Q-type antagonist omega-CgTx MVIIC and by the T-type antagonist amiloride, and is totally inhibited by the N-type antagonist omega-CgTx GVIA. No inhibitory effect is shown by the P-type antagonist omega-Aga TK. We hypothesize that L-type, N-type and T-type VDCCs may be involved to different degrees in the high-K(+)-stimulated JHA release and transient [Ca(2+)](i) increase in the individual CA cells of the adult male M. loreyi, and that the N-type VDCCs may play important roles in these cellular events.