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PubMed1996

Genetic manipulations of microorganisms for the degradation of hexachlorocyclohexane.

Hexachlorocyclohexane (HCH) is an organochlorine insecticide which has been banned in technologically advanced countries. However, it is still in use in tropical countries for mosquito control and thus new areas continue to be contaminated. Anaerobic degradation of HCH isomers have been well documented but until recently there have been only a few reports on aerobic microbial degradation of HCH isomers. The isolation of these microbes made it possible to design experiments for the cloning of the catabolic genes responsible for degradation. We review the microbial degradation of HCH isomers coupled with the genetic manipulations of the catabolic genes. The first part discusses the persistence of residues in the environment and microbial degradation while the second part gives an account of the genetic manipulations of catabolic genes involved in the degradation.

Bacteria↗
PubMed1997

Dengue: A Literature Review and Case Study of Travelers from the United States, 1986-1994.

Dengue fever is an acute, mosquito-transmitted viral disease characterized by fever, headache, arthralgia, myalgia, rash, nausea, and vomiting. Infections are caused by any of four virus serotypes (DEN-1, DEN-2, DEN-3, and DEN-4). The incidence of dengue is increasing in most tropical areas throughout the world (Fig. 1). Although dengue is not endemic in the continental United States, Hawaii, or Alaska, more than 500 laboratory-positive cases of introduced dengue were reported from 1977 through 1994 in U.S. residents who visited dengue-endemic areas throughout the world.1-4 In addition, two competent mosquito vectors (Aedes aegypti and Aedes albopictus) are found in the southeastern United States, and both could possibly transmit an introduced virus. In Hawaii, Ae. albopictus is the dominant mosquito on all islands; Ae. aegypti has only focal distribution on Molokai and the Kona coast of Hawaii. Economic, political, technologic, ecologic, and demographic changes have brought about the emergence of new microbial diseases, as well as an increase in the incidence of previously known infections. The increase in dengue activity in Asia, Africa, and the Americas represents a pandemic that is being facilitated by increased air travel; global urbanization; population growth; greater abundance of disposable, nondegradable containers that can serve as Aedes production sites; and lack of effective mosquito control programs.5,6 This report summarizes information about risk factors for severe disease, recent dengue outbreaks throughout the world, and cases of dengue virus infection in travelers who have been diagnosed on return to the United States.

Journal Article↗
PubMed2001

Vector surveillance for West Nile virus.

West Nile virus (WNV) was detected in the metropolitan New York City (NYC) area during the summer and fall of 1999. Sixty-two human cases, including seven fatalities, were documented. The New York State Department of Health (NYSDOH) initiated and implemented a statewide mosquito and WNV surveillance system. We developed a WNV response plan designed to provide local health departments (LHD) a standardized means to begin to assess basic mosquito population data and to detect WNV circulation in mosquito populations. During the 2000 arbovirus surveillance season, local health agencies collected 317,676 mosquitoes and submitted 9,952 pools for virus testing. NYSDOH polymerase chain reaction (PCR) testing detected 363 WNV-positive pools. Eight species of mosquitoes were found to be infected. Of the 26 counties conducting mosquito surveillance, WNV-positive mosquitoes were detected only in NYC, on Long Island, and in four counties in the lower Hudson River valley region. LHD larval surveillance provided initial or enhanced mosquito habitat location and characterization and mosquito species documentation. Adult mosquito surveillance provided LHD information on species' presence, density, seasonal fluctuations, virus infection, minimum infection ratios (MIR) and indirect data on mosquito control efficacy after larval or adult control interventions. Collective surveillance activities conducted during 1999 and 2000 suggest that WNV has dispersed throughout the state and may affect local health jurisdictions within NYS, adjacent states, and Canada in future years. Vector surveillance will remain a critical component of LHD programs addressing public health concerns related to WNV.

Animals↗
PubMed2006

Natural malaria infection in Anopheles gambiae is regulated by a single genomic control region.

We surveyed an Anopheles gambiae population in a West African malaria transmission zone for naturally occurring genetic loci that control mosquito infection with the human malaria parasite, Plasmodium falciparum. The strongest Plasmodium resistance loci cluster in a small region of chromosome 2L and each locus explains at least 89% of parasite-free mosquitoes in independent pedigrees. Together, the clustered loci form a genomic Plasmodium-resistance island that explains most of the genetic variation for malaria parasite infection of mosquitoes in nature. Among the candidate genes in this chromosome region, RNA interference knockdown assays confirm a role in Plasmodium resistance for Anopheles Plasmodium-responsive leucine-rich repeat 1 (APL1), encoding a leucine-rich repeat protein that is similar to molecules involved in natural pathogen resistance mechanisms in plants and mammals.

Alleles↗
PubMed1983

Toxicity of Bacillus thuringiensis subsp. israelensis to adult Aedes aegypti mosquitoes.

Adult female Aedes aegypti mosquitoes were killed by the parasporal crystals of Bacillus thuringiensis subsp. israelensis (ONR-60A) when the crystals were introduced into the insect midgut as an enema. The 50% lethal dose for intact parasporal crystals was 0.21 microgram/mg of mosquito (wet weight), and for solubilized crystals the 50% lethal dose was 0.04 microgram/mg. These values were compared with 50% lethal concentrations in a free-feeding larval mosquito bioassay of 0.018 and 1.28 microgram/ml for intact and solubilized crystals, respectively. Preparations from B. thuringiensis subsp. kurstaki were ineffective against both adult and larval mosquitoes. An adult mosquito bioassay is suggested as a direct means of screening potential mosquito control agents.

Aedes↗
PubMed2001

Various levels of cross-resistance to Bacillus sphaericus strains in Culex pipiens (Diptera: Culicidae) colonies resistant to B. sphaericus strain 2362.

We studied the cross-resistance to three highly toxic Bacillus sphaericus strains, IAB-59 (serotype H6), IAB-881 (serotype H3), and IAB-872 (serotype H48), of four colonies of the Culex pipiens complex resistant to B. sphaericus 2362 and 1593, both of which are serotype H5a5b strains. Two field-selected highly resistant colonies originating from India (KOCHI, 17,000-fold resistance) and France (SPHAE, 23,000-fold resistance) and a highly resistant laboratory-selected colony from California (GeoR, 36,000-fold resistance) showed strong cross-resistance to strains IAB-881 and IAB-872 but significantly weaker cross-resistance to IAB-59 (3- to 43-fold resistance). In contrast, a laboratory-selected California colony with low-level resistance (JRMM-R, 5-fold resistance) displayed similar levels of resistance (5- to 10-fold) to all of the B. sphaericus strains tested. Thus, among the mosquitocidal strains of B. sphaericus we identified a strain, IAB-59, which was toxic to several Culex colonies that were highly resistant to commercial strains 2362 and 1593. Our analysis also indicated that strain IAB-59 may possess other larvicidal factors. These results could have important implications for the development of resistance management strategies for area-wide mosquito control programs based on the use of B. sphaericus preparations.

Animals↗
PubMed2001

West Nile virus recombinant DNA vaccine protects mouse and horse from virus challenge and expresses in vitro a noninfectious recombinant antigen that can be used in enzyme-linked immunosorbent assays.

Introduction of West Nile (WN) virus into the United States in 1999 created major human and animal health concerns. Currently, no human or veterinary vaccine is available to prevent WN viral infection, and mosquito control is the only practical strategy to combat the spread of disease. Starting with a previously designed eukaryotic expression vector, we constructed a recombinant plasmid (pCBWN) that expressed the WN virus prM and E proteins. A single intramuscular injection of pCBWN DNA induced protective immunity, preventing WN virus infection in mice and horses. Recombinant plasmid-transformed COS-1 cells expressed and secreted high levels of WN virus prM and E proteins into the culture medium. The medium was treated with polyethylene glycol to concentrate proteins. The resultant, containing high-titered recombinant WN virus antigen, proved to be an excellent alternative to the more traditional suckling-mouse brain WN virus antigen used in the immunoglobulin M (IgM) antibody-capture and indirect IgG enzyme-linked immunosorbent assays. This recombinant antigen has great potential to become the antigen of choice and will facilitate the standardization of reagents and implementation of WN virus surveillance in the United States and elsewhere.

Amino Acid Sequence↗
PubMed2005

Differential gene expression in normal human mammary epithelial cells treated with malathion monitored by DNA microarrays.

Organophosphate pesticides are a major source of occupational exposure in the United States. Moreover, malathion has been sprayed over major urban populations in an effort to control mosquitoes carrying West Nile virus. Previous research, reviewed by the U.S. Environmental Protection Agency, on the genotoxicity and carcinogenicity of malathion has been inconclusive, although malathion is a known endocrine disruptor. Here, interindividual variations and commonality of gene expression signatures have been studied in normal human mammary epithelial cells from four women undergoing reduction mammoplasty. The cell strains were obtained from the discarded tissues through the Cooperative Human Tissue Network (sponsors: National Cancer Institute and National Disease Research Interchange). Interindividual variation of gene expression patterns in response to malathion was observed in various clustering patterns for the four cell strains. Further clustering identified three genes with increased expression after treatment in all four cell strains. These genes were two aldo-keto reductases (AKR1C1 and AKR1C2) and an estrogen-responsive gene (EBBP). Decreased expression of six RNA species was seen at various time points in all cell strains analyzed: plasminogen activator (PLAT), centromere protein F (CPF), replication factor C (RFC3), thymidylate synthetase (TYMS), a putative mitotic checkpoint kinase (BUB1), and a gene of unknown function (GenBank accession no. AI859865). Expression changes in all these genes, detected by DNA microarrays, have been verified by real-time polymerase chain reaction. Differential changes in expression of these genes may yield biomarkers that provide insight into interindividual variation in malathion toxicity.

Epithelial Cells↗
PubMed2004

Dengue vaccine: priorities and progress.

Dengue transmission has increased considerably in the past 20 years. Currently, it can only be reduced by mosquito control; however, the application of vector-control methods are labor intensive, require discipline and diligence, and are hard to sustain. In this context, a safe dengue vaccine that confers long-lasting protection against infection with the four dengue viruses is urgently required. This review will discuss the requirements of a dengue vaccine, problems, and advances that have been made. Finally, new targets for research will be presented.

Animals↗
PubMed1997

Survey of cyclopids (Crustacea, Copepoda) in Brazil and preliminary screening of their potential as dengue vector predators.

INTRODUCTION: Cyclopid copepods are known to be good mosquito controllers, specially as regards the larvae of the dengue vectors Aedes aegypti and Ae. albopictus. MATERIAL AND METHOD: The objective of the study was to survey the local copepod fauna and search for new strains of M. longisetus var. longisetus, comparing the potential of the samples found with the current strain ML-01 against Ae. albopictus larvae, under laboratory conditions. Eleven bodies of water in Campinas, SP, Brazil, were screened for copepods by collecting 1.5 l of water from each of then. The predatory potential of adults copepods was evaluated over 24 h, in the laboratory, for groups of 5 individuals preying upon 30 first instar Ae. albopictus larvae. RESULTS AND CONCLUSION: The following cyclopid species were found: Metacyclops mendocinus, Tropocyclops prasinus, Eucyclops sp, Eucyclops serrulatus, Eucyclops solitarius, Eucyclops ensifer, Macrocyclops albidus var. albidus and Mesocyclops longisetus var. longisetus. The predatory potential of these copepods ranged from nil to 97.3%. A sample collected in the field containing only M. longisetus var. longisetus showed the best control efficiency with no significant difference from a three-year old laboratory culture (ML-01) of the same species evaluated for comparison. The sample with few M. albidus var. albidus was ranked in second place showing an average 25.9% efficiency. The use of copepods in trap tires as dengue vector controllers is discussed.

Aedes↗
PubMedDate not supplied

Evaluation of insecticide resistance and biochemical mechanisms in a population of Culex quinquefasciatus (Diptera: Culicidae) from São Paulo, Brazil.

To establish an insecticidal resistance surveillance program, Culex quinquefasciatus mosquitoes from São Paulo, Brazil, were colonized (PIN95 strain) and analyzed for levels of resistance. The PIN95 strain showed low levels of resistance to organophosphates [malathion (3.3-fold), fenitrothion (11.2-fold)] and a carbamate [propoxur (3.0-fold)]. We also observed an increase of 7.4 and 9.9 in alpha and beta esterase activities, respectively, when compared with the reference IAL strain. An alteration in the sensitivity of acetylcholinesterase to insecticide inhibition was also found in the PIN95 mosquitoes. The resistant allele (Ace.1R), however, was found at low frequencies (0.12) and does not play an important role in the described insecticide resistance. One year later, Cx. quinquefasciatus mosquitoes were collected (PIN96 strain) at the same site and compared to the PIN95 strain. The esterase activity patterns observed for the PIN96 strain were similar to those of the PIN95 mosquitoes. However the occurrence of the Ace.1R allele was statistically higher in the PIN96 strain. The results show that esterase-based insecticide resistance was established in the PIN95 Cx. quinquefasciatus population and that an acethylcholinesterase based resistant mechanism has been selected for. A continuous monitoring of this phenomenon is fundamental for rational mosquito control and insecticide application programs.

Animals↗
PubMed2002

Variation of the oviposition preferences of Aedes aegypti in function of substratum and humidity.

Two Aedes aegypti (L.) populations were studied in the laboratory regarding the preference for three types of breeding sites, i.e., flasks containing only water, flasks with a plant and flasks with a stick. Each of these breeding units was placed in one cage and the choice of the oviposition sites was determined for individual females and three females per experimental unit at two humidity levels. Preference for ovipositing on the water surface was observed and varied according to experimental unit and humidity. Mean hatching of eggs in water surface was 46.6%. Experiments with three females showed a more marked difference than when only one female was used. Inter and intrapopulation variability regarding oviposition sites was observed. The discrimination between the different oviposition substrates, hatching in water surface and its implication for mosquito control are discussed.

Aedes↗
PubMed2006

A new black fly isolate of Bacillus thuringiensis autoagglutinating strain highly toxic to Simulium pertinax (Kollar) (Diptera, Simuliidae) larvae.

Formulations containing the entomopathogenic Bacillus thuringiensis serovar israelensis strain IPS-82 has been widely applied for mosquito control around the world. Strain IPS-82 is highly active against Aedes aegypti but less active against other well-known vectors such as Culex quinquefasciatus and Simulium spp. larvae. Eighteen strains of B. thuringiensis were isolated from Simulium pertinax larvae naturally occurring in rivers of Southeast Brazil with one demonstrating special toxic effects. Simulated field tests against S. pertinax larvae showed that the native Brazilian autoagglutinating B. thuringiensis (LFB-FIOCRUZ 1035) has an LC50 at least 25 times lower than the standard IPS-82 strain. The same bacterial preparation was also tested against Ae. aegypti larvae in laboratory trials and the LC50 values obtained with LFB-FIOCRUZ 1035 were at least three times lower than the one for the IPS 82 strain. The results indicate that this strain is more toxic than the standard B. thuringiensis serovar israelensis (H14) in the two Dipteran species tested. It is noteworthy that differences between LC50 values were more pronounced in S. pertinax larvae, the source of the original isolation.

Aedes↗
PubMed2001

Baseline susceptibility to bacterial insecticides in populations of Culex pipiens complex (Diptera: Culicidae) from California and from the Mediterranean Island of Cyprus.

Bacterial insecticides play an increasingly important role in mosquito control. To establish guidelines for detecting resistance at an early stage, information on natural variation in susceptibility of insect populations to these insecticides is needed. Between 1990 and 1993, the susceptibility of Culex pipiens L. complex to Bacillus thuringiensis subsp. israelensis de Barjac and/or Bacillus sphaericus Neide was determined in 31 collections from California. These collections were undertaken before the widespread use of B. thuringiensis subsp. israelensis and before the registration of B. sphaericus in California. Seven collections from the Mediterranean island of Cyprus, where no microbial insecticides have been used, also were tested. The 1990-1991 California collections exhibited limited variation in susceptibility to B. thuringiensis subsp. israelensis. LC50 and LC95 values spanned about a three-fold and four-fold range, respectively. The 1993 Cyprus collections exhibited both higher mean LC values, and greater variability in those values, than the California collections. The LC50s for the Cyprus collections varied over a 10-fold range, whereas the LC50s varied over a 12.5-fold range. Variation in susceptibility to B. sphaericus among the 1991 California collections was about five-fold at the LC50 and LC95. No significant geographic variation in susceptibility to B. thuringiensis subsp. israelensis was observed among regions within California. Although variation in susceptibility was limited among California collections, the greater variability observed among the Cyprus collections and between the Cyprus and California collections illustrates the importance of establishing regional baselines to monitor accurately for changes in susceptibility.

Animals↗
PubMed2006

Laboratory evaluation of pyriproxyfen and spinosad, alone and in combination, against Aedes aegypti larvae.

In this study, the efficacy of pyriproxyfen and spinosad, alone and in combination, was evaluated against the dengue vector Aedes aegypti (L.). Larval bioassays were carried out on susceptible mosquito larvae to determine the concentration-mortality responses of mosquitoes exposed to each insecticide alone and in mixture. Synergism between pyriproxyfen and spinosad was determined by the calculation of a combination index (CI) by using the isobologram method. For pyriproxyfen, LC50 and LC95 were 1.1 x 10(-4) (1.0 x 10(-4)-1.1 x 10(-4)) and 3.2 x 10(-4) (2.9 x 10(-4)-3.6 x 10(-4)) mg/liter, respectively. Pyriproxyfen acted at very low concentrations by inhibiting the adult emergence of Ae. aegypti (97% inhibition rates at 3.3 x 10(-4) mg/liter). Spinosad activity was -500 times lower than that of pyriproxyfen against the Bora strain, with LC50 and LC95 values estimated at 0.055 (0.047-0.064) and 0.20 (0.15-0.27) mg/liter, respectively. A binary mixture of pyriproxyfen and spinosad was realized at the ratio 1:500 by considering the values of the LC50 obtained for each product. The LC50 and LC95 of the mixture were 0.019 (0.016 - 0.022) and 0.050 (0.040 - 0.065) mg/liter, respectively. The mixture combined both the larvicidal activity of spinosad and the juvenoid action of pyriproxyfen. From the LC70 to LC99 a significant synergism effect was observed between the two insecticides (CI ranged from 0.74 to 0.31). This strong synergism observed at high concentrations allows a reduction by five and nine-fold of pyriproxyfen and spinosad amounts to kill almost 100% mosquitoes. Combination of pyriproxyfen and spinosad may then represent a promising strategy to improve mosquito control in situations with insecticide-resistant Aedes dengue vectors.

Aedes↗
PubMed2001

Toxic effect of garlic extracts on the eggs of Aedes aegypti (Diptera: Culicidae): a scanning electron microscopic study.

The surface features of the eggs of Aedes aegypti (L.) and the effect of garlic extracts on their hatching were studied by scanning electron microscopy. The exochorion and endochorion layers of the eggshells display an essentially pentagonal reticulation. The exochorion meshwork exhibits large and small papillae interconnected by horizontal struts. At higher magnification, the large papillae show aeropyles on their rough surface. Eggs hatched in deionized water undergo complete fracture near the anterior poles producing free shell caps. In contrast, eggs placed in 6% reconstituted Kyolic garlic extract are only partially fractured, display attached shell caps, and the larvae remain trapped within the shells. In the natural garlic bulb extract, the eggs show no fracture lines in their shells. No larvae were observed either alive or dead in the garlic extracts, suggesting the embryos were disabled before they could escape from their eggshells as viable larvae. It is concluded that aqueous extracts of garlic inhibit hatching of Aedes eggs. Thus, compounds in garlic may be beneficial in the control of mosquitoes.

Aedes↗
PubMed2002

Cross-resistance to pyrethroid and organophosphorus insecticides induced by selection with temephos in Aedes aegypti (Diptera: Culicidae) from Cuba.

A sample of Aedes aegypti (L.) from Santiago de Cuba, Cuba, with a high level of temephos resistance (19.58x at the 50% lethal concentration [LC50]), was subjected to temephos selection to evaluate the utility of this organophosphate insecticide for mosquito control. High resistance developed after six generations of selection (200.00x). Little or no cross-resistance was observed to the organophosphates, malathion and fenitrothion, but high cross-resistance was observed for the pyrethroid deltamethrin (337.5x) and the organophosphate fenthion (12.74x). Synergism tests implicated detoxifying esterases in temephos and fenthion resistance and deltamethrin resistance was associated with a cytochrome P450 monooxygenasa. Biochemical tests, polyacrylamide gel electrophoresis (PAGE), and inhibition studies confirmed the presence of elevated esterase activity associated with temephos resistance. Pyrethroid resistance could be associated with a glutathione-S-transferase mechanism but not the esterases. The cross-resistance to deltamethrin from temephos selection could limit the use of both insecticides for Ae. aegypti control.

Aedes↗