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Biomedical subjects

R J Novak

Publications and source records attributed to R J Novak.

At least 37 records · Page 2Linked to original sources

Oviposition preferences of Culex pipiens and Culex restuans for infusion-baited traps.

The number of egg rafts oviposited by Culex pipiens and Culex restuans in infusion-baited ovitraps varied with the type of infusion substrate, infusion age, method of infusion preparation, and calendar date. In one or more tests, more Culex egg rafts were collected from water infused with Kentucky bluegrass sod, mixed grass clippings, straw, or a commercial rabbit chow than from a water control. Infusions prepared with oak leaves, maple leaves, and sod soil were ineffective as Culex oviposition attractants; however, alfalfa infusions ranged from marginally effective to unattractive. Rabbit chow infusions required one or more reinfusions before becoming active oviposition lures when prepared in unconditioned (unused) ovitraps. Infusions prepared in conditioned ovitraps (those that had previously contained the same type of infusion) had shorter delay periods. Effective oviposition lures for Culex species included continuous infusions, such as sod and sod grass, and those in which the substrate was infused in the water for only a brief period (24-48 h), such as infusions of rabbit chow and mixed grass clippings. The percentage of egg rafts from Cx. restuans was usually greater in sod and grass infusions than in rabbit chow infusions, whereas Cx. pipiens showed a slight preference for rabbit chow infusions over sod and grass infusions. The temporal shift in relative oviposition activity of the 2 Culex species (cross-over) varied from year to year and between sites in one year.

Animals↗

Attraction of Aedes albopictus adults in sod infusion.

Aedes albopictus adults were recovered from sod-baited gravid traps at 7 sites in 5 counties of southern and southwestern Illinois in areas ranging from suburban to rural. The attraction of this species to a lure usually associated with members of the Culex pipiens complex was investigated under laboratory and field conditions. Olfactometer and oviposition preference tests in the laboratory showed that Ae. albopictus is attracted to sod infusion and females readily oviposit on substrates in contact with the infusion. Both males and females responded to the sod infusion in the olfactometer. The attraction of multiple mosquito species to sod-baited gravid traps was demonstrated at a waste tire site in Jasper County, Illinois. Females of Ae. albopictus, Aedes triseriatus, and Culex species were collected from gravid traps placed along the edge of woods at distances ranging from 100 to 200 m from the tire site. Only a small percentage of the Ae. albopictus females appeared to be gravid (less than 5%), unlike the majority of Culex and Ae. tristeriatus females.

Aedes↗

Dispersal of 10-14-mesh corncob granules in stacked tires.

Dispersal of 10-14-mesh corncob granules was evaluated in 2 random-stacked tire piles, one shingle-stacked tire pile, and one column-stacked tire pile located in a used-tire storage facility in Chicago, IL. Ninety percent and 98%, respectively, of the tires in the 2 random-stacked piles contained granules. In the shingle-stacked tire pile 87% of the tires sampled contained granules, and the number of granules per tire was dependent on depth. The 2 bottom rows of tires were 73.9% less likely to contain granules than the 5 rows above them. In the column-stacked tire pile 91.2% of the tires contained granules and the relationship between granule recovery and tire depth was logarithmic. Overall, the dispersal of 10-14-mesh corncob granules was comparable to that of 8-mesh corncob granules evaluated in a previous study at this site.

Mosquito Control↗

Restriction analysis of the ribosomal DNA internal transcribed spacer region of Culex restuans and mosquitoes in the Culex pipiens complex.

Members of the Culex pipiens Linn. complex in the eastern, southern, and central United States are the primary vectors of St. Louis encephalitis virus. Although species and subspecies in the complex can be identified as 4th-instar larvae and by characters on the male genitalia, adult females cannot be identified accurately. In this study a ribosomal DNA (rDNA) segment that includes the internal transcribed spacer region (ITS) was amplified from Culex pipiens pipiens Linn., Culex quinquefasciatus Say, and Culex restuans Theobald. The DNA was amplified from single abdomens or single legs. The amplified rDNA segment from Cx. restuans is 90 base pairs smaller than those from members of the Cx. pipiens complex. Ribosomal DNA was amplified separately from 3 individuals for each population of Cx. pipiens and analyzed by restriction digestion. Intrapopulation variation is seen, because for each population, bands are present that are common to all 3 individuals within the population, but are also unique to that population. These results indicate that this method may provide a means for distinguishing among the mosquitoes in the Cx. pipiens complex.

Animals↗

Aedes albopictus and Ae. triseriatus eggs survive waste tire processing.

Tires that were either crudely chopped or more finely processed into shreds contained viable eggs. Field-collected remnants of 2-3 chopped tires contained viable Aedes albopictus eggs. After shredding tires seeded with mosquito eggs, 34 (4.6%) of an estimated 746 Ae. albopictus eggs and 21 (2.7%) of an estimated 774 Aedes triseriatus eggs survived. Chopped and shredded tire remnants may serve as a means of dispersing mosquitoes.

Aedes↗

Cellular fatty acid analysis of Bacillus thuringiensis ssp. israelensis (ONR-60A).

The cellular fatty acid composition of Bacillus thuringiensis ssp. israelensis (B.t.i.) ONR-60A was determined by gas-liquid chromatography (GLC). Isolates of ONR-60A were obtained from the Bacillus Genetic Stock Center, Pasteur Institute, and USDA culture collections. These isolates were compared to one another and the culture collections differed in the strains that they contained. Eleven separate strains of ONR-60A were identified, based on fatty acid composition of the cell envelope. The current Pasteur Institute international standard for B.t.i., IPS 82, was also analyzed and it consisted of 3 strains using these same criteria. Profiles were created for each strain found in ONR-60A and IPS 82, and validated against profiles of commercially produced B.t.i. from 2 time periods, 1983-84, and 1990-94. Approximately 84% of the commercially produced B.t.i. (134 out of 160 agar plates) was identified as belonging to the strains found in ONR-60A and in IPS 82. The most prevalent strain found in IPS 82 is also the most prevalent strain in the recently produced commercial material, 40.8% of the commercial plates analyzed, but commercially produced B.t.i. also retained a strong ONR-60A ancestry (35% of the plates analyzed).

Bacillus thuringiensis↗

Mosquito productivity and surveillance for St. Louis encephalitis virus in Chicago during 1993.

The City of Chicago's Department of Health monitors weekly deposition of egg rafts of Culex species, prevalence of St. Louis encephalitis (SLE) virus-specific antibodies in feral birds, and prevalence of the virus in mosquito pools. The total number of Culex egg rafts collected in 1993 (4,623) was 2-fold greater than for the 1992 mosquito season. Virtually all of the early summer egg rafts were identified as Culex restuans. After the week of July 18, Culex pipiens accounted for 20-70% of the total rafts collected weekly. The prevalences for SLE viral antibodies (avian) and RNA (mosquitoes) were 0.2% and 0.02%, respectively. Both values were about 25-fold less than normally occur in epidemic years. It is important for practical considerations to continue this surveillance in order to recommend time- and site-specific mosquito abatement.

Animals↗

Notes on the biology of Orthopodomyia in Illinois.

The principal habitats of immature Orthopodomyia alba and Orthopodomyia signifera are tree holes, though they are occasionally encountered in artificial containers. We collected 929 Or. alba and 17 Or. signifera from 2 plastic trash cans, 2 scrap tires, and a tree hole. All collections were made in an urban residential area in central Illinois.

Animals↗

A North American model to contain the spread of Aedes albopictus through tire legislation.

The introduction, spread, and establishment of the Asian tiger mosquito, Aedes albopictus, in the United States has been facilitated by used or waste tires. This species has not only caused considerable concern among public health officials but also highlighted that the growing accumulations of used tires are a serious solid waste problem and a challenge to the recycling industry. This paper illustrates how the State of Illinois addressed the public health and solid waste problems associated with waste tires, mosquitoes and mosquito-borne pathogens. The goal of the State was to develop and implement through legislative process the Illinois Waste Tire Act. The purpose of the Waste Tire Act is: 1) to ensure that used and waste tires are collected for disposal and recycling, 2) to provide for the abatement of used and waste tire dumps and associated threats to public health, 3) to encourage the development of used and waste tire processing facilities and technologies, including energy recovery, and 4) to provide for research on vectors associated with used and waste tires, and the diseases they spread. The discussion will include key statutes from the Act and the model used to coordinate the effort of five different governmental agencies. An up-to-date presentation of vector-related research, mandated by the Act, ranging from applied investigations on mosquito control to more basic studies investigations on virus identification using molecular technology will be discussed. Finally a discussion of problems that have been encountered along with solutions dealing with legislative language, multidisciplined agency efforts and interactions with the general public are presented.

Adaptation, Physiological↗

PCR-based detection of arboviral RNA from mosquitoes homogenized in detergent.

An improved method for the extraction of viral RNAs was developed to facilitate the reverse transcription (RT)-PCR detection of mosquitoes infected with Western equine encephalitis virus or La Crosse virus. The solubilization method, which uses only EDTA and sodium dodecyl sulfate (SDS) followed by dilution of sample, allows accurate viral detection through the use of random hexamers for the RT followed by specific primers for the PCR. Identities of the reaction products were confirmed either by sequencing or restriction endonuclease digestion. Previous methods for the extraction of RNA for the coupled RT-PCR depended on combinations of guanidinium isothiocyanate, acid phenol, detergents and multiple centrifugations. Ideally, routine detection of viral RNAs for diagnostic purposes should bypass many of the above steps, while still providing a sensitive assay. Our level of detection is 1 infected mosquito in a group of 100.

Animals↗

A rapid diagnostic assay for eastern equine encephalomyelitis viral RNA.

Eastern equine encephalomyelitis virus (EEEV) has been a low-frequency, but serious human and veterinary health problem. Increased frequency of this mosquito-borne virus is anticipated as wetlands are maintained and re-established. Control of EEEV has depended on mosquito abatement in response to increasing frequency of EEEV in the environment. A coupled reverse transcription/polymerase chain reaction assay was designed to rapidly, sensitively, and specifically detect EEEV RNA. The assay successfully detected the viral RNA in a single-blind study of a set of field samples composed of either pooled mosquitoes or bird tissue. These results suggest that it would be practical to use this assay for deciding when and where to implement mosquito abatement.

Animals↗

Use of cellular fatty acid analysis to characterize commercial brands of Bacillus thuringiensis var. israelensis.

The cellular fatty acid composition of Bacillus thuringiensis var. israelensis (B.t.i.) from 5 commercial brands (Vectobac, Acrobe, Skeetal, Bactimos and Teknar), as well as of the current International Standard for B.t.i. (IPS 82), was determined using a Hewlett-Packard Microbial Identification System. The original strain of B.t.i., B.t. var. kurstaki, B.t. var. thuringiensis, B.t. var. morrisoni and Bacillus sphaericus strain 2362 were used as outgroups. Acrobe, Bactimos, Teknar, Vectobac and IPS 82 consisted of the same strain. Skeetal represented a different strain than the other commercially produced B.t.i. Our results indicate that cellular fatty acid analysis can be used to distinguish among the forms of B.t.i. produced by various manufacturers.

Bacillus thuringiensis↗

Reducing the overwintering ability of Aedes albopictus by male release.

Eggs of temperate Aedes albopictus populations are cold hardy and can diapause, but tropical populations are not cold hardy and cannot diapause. Heterozygotes possess intermediate diapause and cold hardiness. Males of a tropical strain from Malaysia with a distinctive genetic marker were released into an existing temperate population in East St. Louis, Illinois. Subsequent egg samples from the release site had genetic marker frequency of up to 24%. Reduced cold hardiness and decreased diapause incidence were also observed in the release site population. No such changes occurred at a nearby control site. The rank order of overwintering survival of eggs at the release site was: Aedes triseriatus > temperate Ae. albopictus > hybrid temperate/tropical Ae. albopictus > tropical Ae. albopictus. Eggs collected from the release population the next summer showed total absence of the genetic marker; presumably carriers were removed by the winter.

Aedes↗

Use of the polymerase chain reaction for the sensitive detection of St. Louis encephalitis viral RNA.

Polymerase chain reaction (PCR) assays were developed for the detection of RNA from the St. Louis encephalitis (SLE) virus. Using computer-assisted analysis of the MSI-7 strain SLE virus genome, two primer pairs were selected from the capsid-coding and the membrane associated protein-coding genes, and one from the envelope-coding gene. Reverse transcription was primed with either specific oligomers or with random hexamers; these methods were compared for cDNA synthesis and subsequent PCR amplification with the oligomeric pairs. Random hexamers provided more sensitive detection of viral RNA. Each primer pair specifically amplified the expected sized fragment from the Parton SLE strain grown in Aedes albopictus cells, but did not amplify Aedes albopictus cell RNA controls. The technique also detected SLE virus RNA in 1 pg of total cellular RNA added to a background of 1 microgram boiled brain tissue, and in 0.5 pg of total RNA added to homogenized mosquito abdomen. PCR-based assays may be adaptable to detect SLE virus RNA in naturally infected mosquitoes, birds, and human cerebrospinal fluid and brain.

Animals↗

Statistical appraisal of the weight-wing length relationship of mosquitoes.

The relationship between wing length and body weight of mosquitoes was examined by comparing the fit of regressions using logarithmic and cubic transformations. A two-parameter line using a double logarithmic transformation provided the best fit, and we found no evidence to justify the use of wing length cubed as a transformation. Wing length was not directly proportional to body weight, but rather increased at a lesser rate than did body weight. Equations that predict body weight from wing length may not be an appropriate substitute for weighing mosquitoes, when studying the extremes of size in populations.

Aedes↗

Effects of Ascogregarina barretti (Eugregarinida: Lecudinidae) infection on Aedes triseriatus (Diptera: Culicidae) in Illinois.

Ascogregarina barretti (Vavra), a gregarine protozoan parasite, infected 23.6 and 5.0% of Aedes triseriatus (Say) adults that emerged from pupae collected from an east-central Illinois tire dump in 1990 and 1991, respectively. Infection significantly reduced the wing length of males and females in both years, but its effect was greatest in 1991. Females were approximately 1.7 times as likely to be infected as males. Infected pupae were 3.53 and 2.76 times more likely to die than uninfected pupae in 1990 and 1991, respectively. Wing length also was affected by location within the tire dump and by tire water volume. We conclude that the A. barretti infection was deleterious to Ae. triseriatus and that the effects of this pathogen may be moderated by environmental factors.

Aedes↗

Detection of St. Louis encephalitis virus in mosquitoes by use of the polymerase chain reaction.

We recently developed an assay using the polymerase chain reaction (PCR) for the specific detection of St. Louis encephalitis (SLE) virus RNA. This assay was tested in a blind study on 7 samples of pooled mosquitoes (50 mosquitoes/pool) which were also characterized for SLE virus by plaque assay in Vero cell culture. One sample was positive for the SLE virus as determined by both the PCR assay and a combination of the plaque assay and the indirect fluorescent antibody assay. The remaining 6 samples were negative for the presence of SLE virus as determined by both methods. These data indicate that this PCR assay can be used to monitor for the presence of SLE virus in pools of homogenized mosquitoes. This approach could provide early data on which to base disease control decisions.

Animals↗

A new method for monitoring mosquito oviposition in artificial and natural containers.

A new method is described for the direct measurement of Aedes albopictus, Ae. atropalpus, Ae. triseriatus and Orthopodomyia signifera oviposition in artificial and natural containers. Seed germination papers were mounted on the insides of can-traps, tree holes and discarded tires to obtain a direct measure of mosquito oviposition. This durable paper substrate was removed weekly while minimizing the disruption of the oviposition habitat. After the germination papers were dried, the aforementioned mosquito eggs were hatched successfully and reared in the laboratory through the adult stage.

Aedes↗