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PubMed2006

Analysis of and function predictions for previously conserved hypothetical or putative proteins in Blochmannia floridanus.

BACKGROUND: There is an increasing interest to better understand endosymbiont capabilities in insects both from an ecological point of view and for pest control. Blochmannia floridanus provides important nutrients for its host, the ant Camponotus, while the bacterium in return is provided with a niche to proliferate. Blochmannia floridanus proteins and metabolites are difficult to study due to its endosymbiontic life style; however, its complete genome sequence became recently available. RESULTS: Improved sequence analysis algorithms, databanks and gene and pathway context methods allowed us to reveal new information on various enzyme and pathways from the Blochmannia floridanus genome sequence [EMBL-ID BX248583]. Furthermore, these predictions are supported and linked to experimental data for instance from structural genomics projects (e.g. Bfl341, Bfl 499) or available biochemical data on proteins from other species which we show here to be related. We were able to assign a confirmed or at least a putative molecular function for 21 from 27 previously conserved hypothetical proteins. For 48 proteins of 66 with a previous putative assignment the function was further clarified. Several of these proteins occur in many proteobacteria and are found to be conserved even in the compact genome of this endosymbiont. To extend and re-test predictions and links to experimentally verified protein functions, functional clusters and interactions were assembled. These included septum initiation and cell division (Bfl165, Bfl303, Bfl248 et al.); translation; transport; the ubiquinone (Bfl547 et al.), the inositol and nitrogen pathways. CONCLUSION: Taken together, our data allow a better and more complete description of the pathway capabilities and life style of this typical endosymbiont.

Animals↗
PubMed2005

Nymphal RNAi: systemic RNAi mediated gene knockdown in juvenile grasshopper.

BACKGROUND: Grasshopper serves as important model system in neuroscience, development and evolution. Representatives of this primitive insect group are also highly relevant targets of pest control efforts. Unfortunately, the lack of genetics or gene specific molecular manipulation imposes major limitations to the study of grasshopper biology. RESULTS: We investigated whether juvenile instars of the grasshopper species Schistocerca americana are conducive to gene silencing via the systemic RNAi pathway. Injection of dsRNA corresponding to the eye colour gene vermilion into first instar nymphs triggered suppression of ommochrome formation in the eye lasting through two instars equivalent to 10-14 days in absolute time. QRT-PCR analysis revealed a two fold decrease of target transcript levels in affected animals. Control injections of EGFP dsRNA did not result in detectable phenotypic changes. RT-PCR and in situ hybridization detected ubiquitous expression of the grasshopper homolog of the dsRNA channel protein gene sid-1 in embryos, nymphs and adults. CONCLUSION: Our results demonstrate that systemic dsRNA application elicits specific and long-term gene silencing in juvenile grasshopper instars. The conservation of systemic RNAi in the grasshopper suggests that this pathway can be exploited for gene specific manipulation of juvenile and adult instars in a wide range of primitive insects.

Animals↗
PubMed2006

Insulated piggyBac vectors for insect transgenesis.

BACKGROUND: Germ-line transformation of insects is now a widely used method for analyzing gene function and for the development of genetically modified strains suitable for pest control programs. The most widely used transposable element for the germ-line transformation of insects is piggyBac. The site of integration of the transgene can influence gene expression due to the effects of nearby transcription enhancers or silent heterochromatic regions. Position effects can be minimized by flanking a transgene with insulator elements. The scs/scs' and gypsy insulators from Drosophila melanogaster as well as the chicken beta-globin HS4 insulator function in both Drosophila and mammalian cells. RESULTS: To minimize position effects we have created a set of piggyBac transformation vectors that contain either the scs/scs', gypsy or chicken beta-globin HS4 insulators. The vectors contain either fluorescent protein or eye color marker genes and have been successfully used for germ-line transformation of Drosophila melanogaster. A set of the scs/scs' vectors contains the coral reef fluorescent protein marker genes AmCyan, ZsGreen and DsRed that have not been optimized for translation in human cells. These marker genes are controlled by a combined GMR-3xP3 enhancer/promoter that gives particularly strong expression in the eyes. This is also the first report of the use of the ZsGreen and AmCyan reef fluorescent proteins as transformation markers in insects. CONCLUSION: The insulated piggyBac vectors should protect transgenes against position effects and thus facilitate fine control of gene expression in a wide spectrum of insect species. These vectors may also be used for transgenesis in other invertebrate species.

Animals↗
PubMed2003

Infection of malaria (Anopheles gambiae s.s.) and filariasis (Culex quinquefasciatus) vectors with the entomopathogenic fungus Metarhizium anisopliae.

BACKGROUND: Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus against tsetse flies (Diptera: Glossinidae) we investigated the potency of this fungus as a biological control agent for adult malaria and filariasis vector mosquitoes. METHODS: In the laboratory, both sexes of Anopheles gambiae sensu stricto and Culex quinquefasciatus were passively contaminated with dry conidia of Metarhizium anisopliae. Pathogenicity of this fungus for An. gambiae was further tested for varying exposure times and different doses of oil-formulated conidia. RESULTS: Comparison of Gompertz survival curves and LT50 values for treated and untreated specimens showed that, for both species, infected mosquitoes died significantly earlier (p < 0.0001) than uninfected control groups. No differences in LT50 values were found for different exposure times (24, 48 hrs or continuous exposure) of An. gambiae to dry conidia. Exposure to oil-formulated conidia (doses ranging from 1.6 x 10(7) to 1.6 x 10(10) conidia/m2) gave LT50 values of 9.69 +/- 1.24 (lowest dose) to 5.89 +/- 0.35 days (highest dose), with infection percentages ranging from 4.4-83.7%. CONCLUSION: Our study marks the first to use an entomopathogenic fungus against adult Afrotropical disease vectors. Given its high pathogenicity for both adult Anopheles and Culex mosquitoes we recommend development of novel targeted indoor application methods for the control of endophagic host-seeking females.

Age Factors↗
PubMed2006

Calcium gluconate and sodium succinate for therapy of sodium fluoroacetate experimental intoxication in cats: clinical and electrocardiographic evaluation.

Sodium fluoroacetate (SFAC) or Compound 1080 is a potent rodenticide, largely used after 1946 for rodent and home pest control. The toxic effects of SFAC are caused by fluorocitrate action, a toxic metabolite, which has a competitive action with aconitase enzyme, leading to citrate accumulation and resulting in interference in energy production by Krebs cycle blockade. In the present study, domestic cats were intoxicated with oral doses of fluoroacetate (0.45 mg/kg). The intoxicated animals presented emesis, diarrhea with abdominal pain posture and an abdominal palpation, tachypnea, bilateral midriasis, hypothermia, hyperexcitability and convulsions. Blood gas analysis indicated decreased pH and bicarbonate levels. Serum ionized calcium was also decreased. ECG showed non-specific changes in ventricular repolarization and ventricular arrhythmias. The survival rate was 75% in the treated group with calcium gluconate and sodium succinate and 37.5% in the nontreated group.

Animals↗
PubMed2002

Implementation of a feral cat management program on a university campus.

In August 1998, Texas AM University implemented on campus a trap-test-vaccinate-alter-return-monitor (TTVARM) program to manage the feral cat population. TTVARM is an internationally recognized term for trapping and neutering programs aimed at management of feral cat populations. In this article we summarize results of the program for the period August 1998 to July 2000. In surgery laboratories, senior veterinary students examined cats that were humanely trapped once a month and tested them for feline leukemia and feline immunodeficiency virus infections, vaccinated, and surgically neutered them. They euthanized cats testing positive for either infectious disease. Volunteers provided food and observed the cats that were returned to their capture sites on campus and maintained in managed colonies. The program placed kittens and tame cats for adoption; cats totaled 158. Of the majority of 158 captured cats, there were less kittens caught in Year 2 than in Year 1. The proportion of tame cats trapped was significantly greater in Year 2 than in Year 1. The prevalence found for feline leukemia and feline immunodeficiency virus ELISA test positives was 5.8% and 6.5%, respectively. Following surgery, 101 cats returned to campus. The project recaptured, retested, and revaccinated more than one-fourth of the cats due for their annual vaccinations. The program placed 32 kittens, juveniles, and tame adults for adoption. The number of cat complaints received by the university's pest control service decreased from Year 1 to Year 2.

Animal Welfare↗
PubMed2001

Targeting of an expressed neurotoxin by its recombinant baculovirus.

AaIT, an insect-selective neurotoxic polypeptide derived from scorpion venom, has recently been used to engineer recombinant baculoviruses for insect pest control. Lepidopterous larvae infected with an AaIT-expressing baculovirus reveal symptoms of paralysis identical to those induced by injection of the native toxin. However, the paralyzed larvae treated by the recombinant virus possess an approximately 50-fold lower hemolymph toxin concentration than insects paralyzed by the native toxin. The mechanism of this potentiation effect was studied using immunocytochemistry, electrophysiology and toxicity assays. (i) Light microscopy, using peroxidase-conjugated antibodies, revealed the presence of toxin in virus-susceptible tissues, including tracheal epithelia located close to the central nervous system and beyond its lamellar enveloping sheath. (ii) High-resolution immunogold electron microscopical cytochemistry clearly revealed the presence of recombinant AaIT toxin inside the thoracic and abdominal ganglia on neuronal cell bodies and axonal membranes. (iii) Ventral nerve cords dissected from silkworm larvae infected with the recombinant baculovirus exhibited a high degree of excitability, expressed as enhanced frequency and bursting mode of their spontaneous activity, when compared to nerve cords infected with the wild-type virus. We conclude that the recombinant-virus-infected tracheal epithelia, outbranching in the body of an infected insect, (i) locally supply a continuous, freshly produced toxin to its neuronal receptors and (ii) introduce the expressed toxin to the insect central nervous system, thus providing it with critical target sites that are inaccessible to the native toxin.

Animals↗
PubMed2005

Potential contamination of forensic entomology samples collected in the mortuary: a case report.

Carrion insects originating from infested bodies may establish small mortuary populations. Two Victorian Institute of Forensic Medicine cases are presented to illustrate how these insects can potentially contaminate forensic entomology samples collected in the mortuary. The first case proves that bodies can be colonised in the mortuary: when a decomposed body in freezer storage was thawed for a procedure in the main mortuary, it was colonised within five minutes of exposure by the beetle Necrobia rufipes De Geer (Cleridae, Coleoptera). The second case illustrates that immobile juvenile insects can be transferred between bodies, and describes the transfer of an Eristalis sp. (Syrphidae, Diptera) pupa from a decomposed to a fresh body. While the contamination risk is slight, it can be further minimised by checking mortuary-collected samples against those taken from the body discovery site, by mortuary pest control, and by checking previous mortuary admittance records for possible contamination sources.

Animals↗
PubMed2000

Biologic monitoring of exposure to organophosphorus pesticides in 195 Italian children.

One hundred ninety-five 6- to 7-year-old children who lived in the municipality of Siena (Tuscany, Italy), underwent biologic monitoring to evaluate urinary excretion of several alkylphosphates that are metabolites of organophosphorus pesticides. We evaluated dimethylphosphate (DMP), dimethylthiophosphate (DMTP), dimethyldithiophosphate (DMDTP), diethylphosphate (DEP), diethylthiophosphate (DETP), and diethyldithiophosphate (DEDTP). We obtained urine samples taken in the children's schools, and each sample was accompanied by a questionnaire about lifestyle and dietary habits. We found DMP and DMTP in detectable concentrations in the greatest number of samples (96 and 94%, respectively). The DMP values were geometric mean (GM) 116.7, [geometric standard deviation (GSD) 2.5], and a range of 7.4-1,471.5 nmol/g creatinine. The corresponding DMTP values were GM 104.3 (GSD 2.8) and a range of 4.0-1,526.0 nmol/g creatinine. DMDTP, DEP, DETP, and DEDTP concentrations were GM 14.1, (GSD 3.0), and a range of 3.3-754.6 nmol/g creatinine in 34% of the children; GM 33.2, (GSD 2.4), and a range of 5.1-360.1 nmol/g creatinine in 75% of the children; GM 16.0, (GSD 2.9), and a range of 3.1-284.7 in 48% of the children; and GM 7.7, (GSD 2.1), and a range of 2.3-140.1 in 12% of the children, respectively. The significant variable for urinary excretion of these metabolites in children was pest control operations performed inside or outside the house in the preceding month; however, the presence of a vegetable garden near the house rarely emerged. The urinary excretion of alkylphosphates in children was significantly higher than in a group of the adult population resident in the same province.

Agriculture↗
PubMed2002

Methods of assessing neurobehavioral development in children exposed to methyl parathion in Mississippi and Ohio.

Methyl parathion (MP), an organophosphate pesticide, was sprayed illegally for pest control in U.S. residences and businesses in Mississippi and Ohio. To evaluate the association between MP exposure and neurobehavioral development, children 6 years of age or younger at the time of the spraying and local comparison groups of unexposed children were assessed using the pediatric environmental neurobehavioral test battery (PENTB). The PENTB is composed of informant-based procedures (parent interview and questionnaires) and performance-based procedures (neurobehavioral tests for children 4 years of age or older) that evaluate each of the four broad domains (cognitive, motor, sensory, and affect) essential to neurobehavioral assessment. Children were classified as exposed or unexposed using urinary p-nitrophenol (PNP) levels and environmental wipe samples for MP. Exposure was defined as a urinary PNP level of greater than or equal to 100 ppb for the child or any other individual living in the household. Environmental wipe sample levels of greater than or equal to 150 g MP/100 cm2 and greater than or equal to 132.9 g MP/100 cm2 were used to define MP exposure for children living in Mississippi and Ohio, respectively. The PENTB was first administered in summer 1999 (year 1). The PENTB was readministered in summer 2000 (year 2) to children who participated in year 1 of the study. A description of the methods used in the study are presented. Results of data analyses for both years of the study will be presented in a separate publication.

Child↗
PubMed2004

Long-term neurobehavioral health effects of methyl parathion exposure in children in Mississippi and Ohio.

Methyl parathion (MP), an organophosphate pesticide licensed only for agricultural uses, was sprayed illegally for pest control in Mississippi and Ohio residences. To evaluate the association between MP exposure and neurobehavioral development, we assessed children 6 years or younger at the time of the spraying and local comparison groups of unexposed children using the Pediatric Environmental Neurobehavioral Test Battery (PENTB). The PENTB is composed of informant-based procedures (parent interview and questionnaires) and performance-based procedures (neurobehavioral tests for children 4 years or older) that evaluate cognitive, motor, sensory, and affect domains essential to neurobehavioral assessment. Children were classified as exposed or unexposed on the basis of urinary para-nitrophenol levels and environmental wipe samples for MP. Exposed children had more difficulties with tasks involving short-term memory and attention. Additionally, parents of exposed children reported that their children had more behavioral and motor skill problems than did parents of unexposed children. However, these effects were not consistently seen at both sites. There were no differences between exposed and unexposed children in tests for general intelligence, the integration of visual and motor skills, and multistep processing. Our findings suggest that MP might be associated with subtle changes to short-term memory and attention and contribute to problems with motor skills and some behaviors, but the results of the study are not conclusive.

Attention↗
PubMed1976

Degradation, metabolism and toxicity of synthetic pyrethroids.

Synthetic pyrethroidal compounds undergo biodegradation in mammals both oxidatively and hydrolytically, and depending on the type of compound, either of the pathways may predominate. Thus, (+) - or (+/-) -trans isomers of the chrysanthemumate ester of primary alcohols such as fenothrin, furamethrin, proparthrin, resmethrin, and tetramethrin (and possibly permethrin, too) are metabolized mainly through hydrolysis of the ester linkage, with subsequent oxidation and/or conjugation of the component alcohol and acid moieties. On the other hand, the corresponding (+)-cis enantiometers and chrysanthemumate of secondary alcohols like allethrin are resistant to hydrolytic attack, and biodegraded via oxidation at various sites of the molecule. These rapid metabolic degradations, together with the presumable incomplete absorption from the gastrointestinal tract, would generally contribute to the low acute toxicity of synthetic pyrethroids. These compounds are neither skin irritants nor skin sensitizers, and inhalation toxicity as well as dermal toxicity are fairly low. Neither is teratogenic in rats, mice, and/or rabbits or mutagenic on various bacterial strains. Subacute and chronic feeding of higher amounts of the compounds to rats invariably causes some histopathological changes in liver; however, these are neither indicative nor suggestive of tumorigenicity. Based on existing toxicological information, the present recommended use patterns might afford sufficient safety margin on human population. However, in extending usage to agricultural pest control, much more extensive investigations should be forthcoming from both chemical and biological aspects, since there is scant information on the fate of these pyrethroids in the environment. Also several of the compounds may be very toxic to certain kinds of fish and arthropods.

Administration, Oral↗
PubMed1976

Health-related aspects of integrated pest management.

Basic concepts and philosophy of integrated pest management are presented in order to dispel several misconceptions and to provide the necessary background information for discussion of its relationship to the health-related effects of pesticide use. Implications for human health of current pesticide practices in Central America are examined to illustrate major problems associated with the injudicious use of insecticides, i.e., human pesticide poisonings, development of insect resistance, and persistence in the environment. Mitigation of these problems would ideally be achieved through the efforts and cooperation of a multidisciplinary team of scientists and technical people in the medical and agricultural sciences. The dilemma associated with the development of integrated pest control systems in developing countries is discussed. The FAO/UNEP Global Programme was reviewed.

Brazil↗
PubMed2006

Measuring potential dermal transfer of a pesticide to children in a child care center.

Currently, the major determinants of children's exposure to pesticides are not fully understood, and approaches for measuring and assessing dermal exposure in a residential setting have not been sufficiently evaluated. In one approach, dermal exposure is estimated using empirically derived transfer coefficients. To assess the feasibility of using this approach for assessing children's exposure to pesticides, we conducted a study was conducted in a child care center that had a preexisting contract with a pest control service for regular monthly pesticide applications. Children in the selected child care center were monitored using full-body cotton garments to measure dermal loading. Pesticide residues on classroom surfaces were measured in the areas where the children spent time. Measured surface-wipe loadings ranged from 0.47 to 120 ng/cm2, and total garment loadings ranged from 0.5 to 660 pg/cm2. The garment and surface loading measurements were used to calculate dermal-transfer coefficients for use in assessing children's residential exposure to pesticides. Dermal-transfer coefficients calculated using these data range from approximately 10 to 6,000 cm2/hr. The wide range in these values demonstrates the importance of developing standard surface-measurement protocols if this approach is to be used to assess dermal exposure in a residential environment. The upper-range values resulting from this study were found to be similar to the default value used by the U.S. Environmental Protection Agency to assess children's dermal exposures resulting from contact with indoor surfaces.

Administration, Cutaneous↗
PubMed1985

Utilization of adipose tissue biopsy in characterizing human halogenated hydrocarbon exposure.

Halogenated hydrocarbons have been successfully utilized for pest control in agriculture and public health. In industry, the polychlorinated biphenyls (PCBs) have been particularly useful. Unfortunately, residues have proven persistent and have been found widely dispersed in the environment. Because they have chemical properties that favor bioaccumulation, it is not surprising that many have been identified in animals and man. Such findings prompted public health concern and initiated review of potential adverse health impacts. In many instances this process has led to total prohibition of use or strict limitations. Highly lipophilic, the primary accumulation site in humans is adipose tissue. Analysis of adipose samples remains the preferred biologic index for estimating exposure. Improved instrumentation has lowered the limit of detection and improved the accuracy of quantification. Accumulated population experience has helped develop understanding of the dynamics of tissue partitioning in humans. Once such relationships have been adequately described, other less invasive procedures may be more confidently used for general surveillance purposes. Whenever adipose tissue is obtained, the blood compartment should also be sampled and the relationship reported. We have only begun to investigate the resulting partition ratio as an investigative tool.

Adipose Tissue↗
PubMed1992

Agricultural chemical utilization and human health.

The public is justifiably concerned about the human health effects of agricultural chemicals. The many gaps in information about the mechanisms of toxic action, human exposures, and the nature and extent of human health effects are large. Very few older pesticides, in particular, have been tested for human health effects. Workers who produce, harvest, store, transport, process, and prepare food and fibers are exposed to many chemicals that are potentially hazardous and that are used in agriculture. The occupational health of these workers has not been adequately studied, and protective efforts have sometimes been minimal. Valid and accurate risk assessment is best based on sound information about how chemicals, in this case agricultural chemicals, are involved in toxic events--their mechanisms of action. These health effects include tumor promotion, chronic and acute neurotoxicity, immunotoxicity, and reproductive and developmental toxicity. Another key part of risk assessment is exposure assessment. Fundamental studies of the toxicology of target organisms and nontarget organisms exposed to agricultural chemicals are needed to discover and develop better solutions to the problems of agricultural pest control, including better formulations, optimal application rates and public education in safety and alternative agricultural practices. The large number of pesticides that have never been adequately tested for effects on human health is particularly worrisome in light of emerging information about delayed nervous system effects.

Agrochemicals↗
PubMed1999

Estrogenic potential of certain pyrethroid compounds in the MCF-7 human breast carcinoma cell line.

Estrogens, whether natural or synthetic, clearly influence reproductive development, senescence, and carcinogenesis. Pyrethroid insecticides are now the most widely used agents for indoor pest control, providing potential for human exposure. Using the MCF-7 human breast carcinoma cell line, we studied the estrogenic potential of several synthetic pyrethroid compounds in vitro using pS2 mRNA levels as the end point. We tested sumithrin, fenvalerate, d-trans allethrin, and permethrin. Nanomolar concentrations of either sumithrin or fenvalerate were sufficient to increase pS2 expression slightly above basal levels. At micromolar concentrations, these two pyrethroid compounds induced pS2 expression to levels comparable to those elicited by 10 nM 17ss-estradiol (fivefold). The estrogenic activity of sumithrin was abolished with co-treatment with an antiestrogen (ICI 164,384), whereas estrogenic activity of fenvalerate was not significantly diminished with antiestrogen co-treatment. In addition, both sumithrin and fenvalerate were able to induce cell proliferation of MCF-7 cells in a dose-response fashion. Neither permethrin nor d-trans allethrin affected pS2 expression. Permethrin had a noticeable effect on cell proliferation at 100 microM, whereas d-trans allethrin slightly induced MCF-7 cell proliferation at 10 microM, but was toxic at higher concentrations. Overall, our studies imply that each pyrethroid compound is unique in its ability to influence several cellular pathways. These findings suggest that pyrethroids should be considered to be hormone disruptors, and their potential to affect endocrine function in humans and wildlife should be investigated.

Breast Neoplasms↗
PubMed1999

Dermal transfer of chlorpyrifos residues from residential surfaces: comparison of hand press, hand drag, wipe, and polyurethane foam roller measurements after broadcast and aerosol pesticide applications.

Indoor residential pesticide applications present the potential for human exposures, particularly for small children. Personal contact with target and nontarget surfaces can result in transfer of pesticides to the skin, but the magnitude of such transfer is uncertain. This research compared surface sampling techniques [wipe and polyurethane foam (PUF) roller] with the removal ability of human skin following broadcast and total aerosol release applications of Dursban (Dow Elanco, Midland, MI), a residential formulation containing the insecticide chlorpyrifos. Hands were washed immediately after surface contact, following a protocol that included a laboratory-generated adjustment factor to account for incomplete removal of chlorpyrifos from skin. Chlorpyrifos transfer was similar for hand press and hand drag techniques, averaging approximately 1-6 ng/cm2 of carpet contacted. These amounts represented < 1% of the amount of chlorpyrifos deposited on the surfaces 3.5 hr earlier. Chlorpyrifos transfer from carpet to skin was 23-24 times lower than for wipe sampling and 33-36 times lower than for PUF roller sampling (p = 0.0007 and p = 0.0006 for broadcast and aerosol applications, respectively). Hand press sampling removed approximately 4.5 times less chlorpyrifos from nontarget furniture surfaces (12 ng/cm2) than did wipe sampling (56 ng/cm2; p = 0.009). Chlorpyrifos residues on carpet were substantially higher after broadcast applications than after aerosol applications, but residues on such nontarget surfaces as furniture were substantially higher for the aerosol application. This study indicates that human skin removes substantially less residue from carpets and furniture than either conventional wipe or PUF roller sampling methods following residential pest control applications of chlorpyrifos. Although this paper focuses on quantifying residue transfer from surface to skin using different surface sampling techniques, no attempt is made to quantify the amount of chlorpyrifos residue that is subsequently absorbed.

Aerosols↗