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Infestation of rodents with larval Ixodes ricinus (Acari: Ixodidae) is an important factor in the transmission cycle of Borrelia burgdorferi s.l. in German woodlands.

The ecology of Borrelia burgdorferi Johnson et al. s.l. was investigated from 1987 to 1993 in a preserved woodland in western Germany near Bonn. In selected biotopes, host-seeking Ixodes ricinus L. were regularly collected by blanket dragging in 1987, 1988, and 1989 and screened for infection with B. burgdorferi. Rodents were trapped monthly between April and October in 1988, 1990, 1991, and in the winter of 1992-1993, examined for antibodies to B. burgdorferi s.l., and inspected for feeding ticks. Ticks collected from rodents were screened for spirochete infection. High numbers of host-seeking nymphs were consistently collected within a biotope characterized by humid and acid soils. The mean number of ticks was significantly lower in biotopes with permeable soils. All small mammals captured belonged to the species Apodemus flavicollis Melchior, A. sylvaticus L., and Clethrionomys glareolus Schreber. Of 11,680 ticks obtained from rodents, 11,674 were I. ricinus, with 97.9% of the ticks being larvae, 2.0% nymphs, and 0.1% females. Mean numbers of feeding ticks ranged from 3.4 to 117 larvae per rodent and from 0.0 to 0.64 nymph per rodent, respectively. High levels of larval infestation on rodents were recorded in the same biotope where high numbers of host-seeking nymphs were present. Members of the genus Apodemus were more heavily infested with I. ricinus larvae than C. glareolus. The mean infection prevalence in host-seeking ticks was found to be 1% for larvae, 5% for nymphs, and 10-20% for adults. The infection prevalence in host-seeking nymphs ranged from 1.1 to 15.4% according to the particular biotope. The values for specific infectivity for the Apodemus populations were positively correlated with the mean larval infestation, but not with nymphal infestation. The respective estimates for C. glareolus were much higher than those for Apodemus spp. in biotopes with low tick densities. However, specific infectivity of C. glareolus was substantially reduced at sites with high tick abundances. In biotopes with high numbers of infected I. ricinus, significantly more rodents were found to have antibodies to B. burgdorferi than in biotopes with low abundances of ticks. The data show that C. glareolus plays a different role as reservoir host species compared with the 2 Apodemus species. This and previous studies suggest that the degree of infestation with larval I. ricinus differentially modulates infectivity of host species for ticks. We conclude that immune processes in natural reservoir hosts induced by B. burgdorferi or I. ricinus bites (or both) are important regulatory factors in the transmission cycle(s) of B. burgdorferi.

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Postembryonic changes in circadian photo-responsiveness rhythms of optic lobe interneurons in the cricket Gryllus bimaculatus.

The photo-responsiveness of 2 groups of interneurons responding to light in the protocerebrum was investigated at 2 developmental stages, the last instar nymphs and adults, in the cricket Gryllus bimaculatus. The cricket is diurnally active during the nymphal stage but becomes nocturnal as an adult. In both adults and nymphs, light-induced responses of optic lobe light-responding interneurons that conduct light information from the optic medulla to the lobula and the cerebral lobe showed a circadian rhythm peaking during the subjective night. Amplitudes of the rhythms were not significantly different between adults and nymphs, but adults showed more stable day and night states than did nymphs. The medulla bilateral neurons that interconnect the bilateral medulla areas of the optic lobe also showed circadian rhythms in their light-induced responses in both adults and nymphs. The rhythm had a clear peak and a trough in adults, and its amplitude was significantly greater than that of nymphs. These results suggest that the 2 classes of interneurons are differentially controlled by the circadian clock. The difference might be related to their functional roles in the animal's circadian behavioral organization.

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[Presence of Triatoma sordida Stäl, 1859 in urban ecotopes, of the city of Corrientes, Argentina].

With the intention of verifying the presence of T. sordida Stál 1859 in urban ecotopes in Corrientes city, Argentina, both, human dwellings and an extradomiciliary habitat were taken as a basis. A complete T. sordida population census was carried on in Mitre Park pigeon house and 400 pigeon nests (Columbia livia) were dissected. The triatomineos samples that had been detected in human dwellings were sent to the Arthropods Laboratory by their inhabitants. These triatomineos were classified systematically and according to their age class. Their feces were examined under a microscope 400 x, so as to identify Trypanosoma cruzi. Six eggs, 45 first-instar nymphs, 110 second-instar nymphs, 226 third-instar nymphs, 303 fourth-instar nymphs, 1.103 fifth-instar nymphs, 12 females and 11 males were collected from the pigeon house. The presence of this species was verified in the 27 houses, situated in 8 different neighborhoods in the city; 13 eggs, 33 different instar nymphs, 27 females and 9 males were captured, 82 samples altogether. No triatomineo was infected by T. cruzi. T. sordida adults moved towards the houses in Spring and Summer. Of the material found in human dwellings 33% was female. The adaptation of T. sordida to extra-domestic habitats and their tendency to invade human houses has thus been verified. This fact should be taken into account in vector control programs.

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Efficacy of ivermectin against the bloodsucking insect, Rhodnius prolixus (Hemiptera, Triatominae).

Ivermectin (0.2 mg/kg body weight) caused a high mortality in nymphs and adults of Rhodnius prolixus following a single meal in mice sub-cutaneously injected with the drug. This effect was more evident in nymphs of 1st-and 2nd-instar than in older nymphs and adults. Third-instar nymphs presented a high mortality when fed on mice treated with ivermectin 24 and 48 hours previously, while mortality was significantly reduced in nymphs fed on mice treated 72 hours before. Surviving 3rd-instar nymphs did not molt. When adult females were fed once on mice treated for 24 hours with ivermectin there was a considerable reduction in egg production. This inhibition was not reversed by a second feeding on normal mice. We concluded that sub-lethal doses of ivermectin caused toxic effects interfering in the neuro-endocrine control of development and reproduction of this bloodsucking insect.

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Effects of pyriproxyfen on off-host water-balance and survival of adult lone star ticks (Acari: Ixodidae).

Newly engorged nymphs of the lone star tick, Amblyoma americanum (L.), were continuously exposed to 4 microg/cm2 of pyriproxyfen residues in glass vials. Treatment of engorged nymphs (n = 285) resulted in significant molting inhibition, with more than one-fourth (26.7%, n = 76) of nymphs dying before or during ecdysis. Treatment effects were evident among ticks that molted to the adult stage, with 26.7% (n = 76) of females, and 17.9% (n = 51) of males exhibiting moribund physical characteristics (i.e., lethargy; dull, discolored and desiccated cuticles; lacking full locomotor competency). A few molted adult ticks (10 males, four females) were dead upon inspection. Only 11.2% of pyriproxyfen treated, emergent females (n = 32), and 11.5% of treated emergent males (n = 25) from 285 ticks treated as engorged nymphs, exhibited normal physical appearance and possessed a full range of locomotor activity. Treated adult ticks maintained within a desiccating environmental chamber at 0% RH and 23 degrees C, had significantly accelerated whole-body water loss rates in comparison to untreated males and females maintained under the same environmental conditions. Additionally, treated adult ticks maintained under optimal environmental conditions (23 degrees C and >95% RH) sustained 100% mortality within 32 d following assignment to these conditions (or 79 d posttreatment as engorged nymphs), whereas untreated ticks had 0% mortality for the same duration of time. Results demonstrate that continuous exposure of nymphs to pyriproxyfen disrupted molting, and accelerated both whole-body water loss and subsequent mortality among emergent adult ticks.

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Feeding patterns of Triatoma longipennis Usinger (Hemiptera, Reduviidae) in peridomestic habitats of a rural community in Jalisco State, Mexico.

We analyzed triatomine blood feeding patterns to evaluate the role of peridomiciles in Trypanosoma cruzi transmission at the rural village of Tepehuaje de Morelos at Jalisco State, Mexico (1999). A total of 206 bugs were collected in 11 out of 26 households (42.3%). Nymphs predominated in the collections (64.9% of the total). Except for one Triatoma barberi female, a species that belongs to the protracta species complex, all adults were Triatoma longipennis, a species of the phyllosoma complex. Triatomines were exclusively present in peridomestic sites mainly piles of tiles and bricks, and none were found indoors. Overall infection rate was 56.6% and no significant differences (P > 0.05) were observed between nymphs and adults or males and females. Identified blood meals were chicken (29.4%), opossum (20.9%), pig (24.5%), murid (20.9%), dog (3.5%), and armadillo (0.7%). No gut content reacted against anti-human, anti-bovine, anti-rabbit, and anti-cat sera. In contrast to fifth nymphs and adults, 87% of the small nymphs fed on one host, indicating that they are less mobile than other stages. Most fifth nymphs and adults fed on domestic hosts, while small nymphs mainly fed on opossum and murid. Infection blood-meal indexes were around 50% for single meals on opossum and murid, stressing their importance as trypanosome donors. Peridomiciles in Tepehuaje could be regarded as interaction sites among domestic and wild and synanthropic mammals and triatomines, which would facilitate circulation of the same T. cruzi strains between domestic and sylvatic cycles. Stone-made walls and building materials, which hold synanthropic rodents and opossums, should be considered as targets for vector control measures.

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Experimental studies on the transmission cycle of Thogoto virus, a candidate orthomyxovirus, in Rhipicephalus appendiculatus.

Thogoto (THO) virus, a candidate orthomyxovirus, replicated in and was transmitted by larvae, nymphs, and adults of the brown ear tick, Rhipicephalus appendiculatus. Larvae fed on viremic hamsters (10(7-8) PFU/ml blood) acquired an average of 10(2.5) PFU per tick. Following engorgement the titer dropped to 10(1.9) PFU on day 2 but increased by day 6 to 10(3.3) PFU. Virus survived transstadially in these ticks as demonstrated by the fact that, on day 10, newly moulted nymphs contained, on average, 10(3.5) PFU/tick. When 10 such infected nymphs were placed on a hamster a fatal infection of the animal developed involving a viremia of 10(6.7) PFU/ml blood. Another group of 6 infected nymphs did not elicit a detectable viremia in a hamster, or cause death. However the animal seroconverted to THO, virus indicating that virus transmission had occurred. Following acquisition of THO virus at the larval stage, virus was detected in adult ticks 138 days later. Uninfected nymphs fed on viremic hamsters acquired an average of 10(4) PFU/nymph. No virus was detected in the nymphs 4 days post-engorgement. Virus was, however, recovered by 6 days post-engorgement (10(4.7) PFU/nymph). Virus persisted transstadially as shown by the presence of an average of 10(3.4) PFU in newly moulted adults. Three groups of these infected adults (5-6 ticks/group) induced viremia in hamsters with blood titers of the order 10(2.8-3.5) PFU/ml. Virus persisted in engorged adults for up to 66 days following nymphal engorgement.(ABSTRACT TRUNCATED AT 250 WORDS)

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Genetic engineering of Periplaneta fuliginosa densovirus as an improved biopesticide.

The smoky-brown cockroach (Periplaneta fuliginosa) densovirus (PfDNV) has previously shown potential in urban pest control. To improve its efficacy as a biopesticide, the genome of PfDNV was engineered by inserting the insect-specific toxin gene BmKIT1 in the open reading frame encoding the major structural proteins. A green fluorescent protein (GFP) marker was tagged to the BmKIT1 at its C-terminus for in vivo imaging using Confocal laser scanning microscopy (CFSM). Using a virion rescue strategy, the genomes of recombinant and wild-type (wt) PfDNV were then cotransfected in P. fuliginosa nymphs. Reverse transcription PCR (RT-PCR) showed that the inserted BmkIT1 genes were expressed in the P. fuliginosa nymphs 48 h after cotransfection. CFSM and transmission electron microscopy also confirmed the generation of virus particles and expression of BmKIT1-GFP fusion protein in the cotransfected nymphs. The recombinant viruses remained infective to P. fuliginosa nymphs in feeding tests. Using the LT(50) bioassay method, the coninfection of the recombinant and wt PfDNV killed the P. fuliginosa nymphs approximate 32% faster than wt PfDNV only. This is the first report showing the improvement of engineered densovirus for the potential application of biopesticide.

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Nymphal Ixodes dammini: models of the temporal abundance patterns.

A difference equation model was developed to explore the sensitivity of the temporal pattern of relative abundance of active, host-seeking nymphal Ixodes dammini. Inputs to the model were the temporal patterns of recruitment of nymphs into the active class, mortality and successful acquisition of hosts by the ticks. Input parameters were varied both in the temporal pattern (shape) and in the cumulative level (summed over the period of activity). The output of the models, the temporal abundance pattern, was examined for (1) overall shape and (2) the timing of the peak abundance of host-seeking nymphs. The shape of the temporal pattern of nymphal abundance was not sensitive to changes in the shape of the functions used as input for host-finding. The time of peak abundance of nymphs is slightly sensitive to changes in the overall level of host-finding and mortality. When more ticks are removed from the active class (host-finding or mortality increasing), the time of the peak abundance of the nymphs shifts earlier. However, this shift is small compared to variation in field data. The general shape of the activity pattern was sensitive to changes in the temporal pattern of recruitment. A left-skewed distribution produced output which most resembles field data. The temporal pattern of nymphs entering the active class is important, and is an area which needs further empirical work.

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Experiments on transmission of an unidentified Theileria sp. to small ruminants with Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum.

Experiments on the transmission of an unidentified Theileria sp. infective for small ruminants by Haemaphysalis qinghaiensis and Hyalomma anatolicum anatolicum were carried out. Three Theileria-free batches of adult, larvae, and nymphs of laboratory reared H. qinghaiensis and Hy. a. anatolicum ticks were infected by feeding them on sheep infected with Theileria sp. The Theileria sp. was originally isolated from adult ticks of H. qinghaiensis, by inoculation of blood stabilates or tick transmission. H. qinghaiensis has been shown to be capable of transmitting the Theileria sp. infective for small ruminants transstadially to sheep and goats. The nymphs developed from the larvae engorged on the sheep infected with the parasite transmitted the pathogen to splenectomized sheep with prepatent periods of 30, 31 days, respectively; but the subsequent adult ticks of H. qinghaiensis derived from the nymphs did not transmit the pathogen to sheep. However, adults developed from the nymphs engorged on the sheep infected with the parasite transmitted the pathogen to sheep with prepatent periods of 24-27 days. The larvae, nymphs and adult ticks derived from female H. qinghaiensis ticks engorged on infected sheep were not able to transmit the parasite transovarially. The same experiments were done with Hy. a. anatolicum, but examination for presence of piroplasma of Theileria sp. from all animals were negative, demonstrating that Hy. a. anatolicum could not transmit the organism to sheep or goats.

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Annual and seasonal variation of tick-borne encephalitis virus (TBEV) prevalence in ticks in selected hot spot areas in Germany using a nRT-PCR: results from 1997 and 1998.

The prevalence of tick-borne encephalitis virus (TBEV) in Ixodes ricinus tick populations in endemic areas of Germany with the highest TBE risk is unknown. Annual and seasonal differences in TBEV prevalence have also not been studied. Against this background, in May 1997 we started a systematic virus surveillance programme in ticks collected in locations known to have a high incidence of autochthonous TBE cases. These were 5 locations in Baden-Württemberg (Black Forest) and 8 locations in Bavaria (surrounding Passau). Field-collected ticks were randomly assigned to pools of 10 adults or 20 nymphs, respectively. The tick pools were tested for the presence of TBEV-RNA using a newly developed, sensitive nested reverse transcriptase polymerase chain reaction assay (nRT-PCR). The primer pairs were selected from the 5'-terminal noncoding region, a highly conserved part of the virus. The specificity was tested by computer homology searches of sequences, as well as by sequencing of the first and the second amplificates, by Southern blot hybridisation with a DIG-labelled oligonucleotide probe, and by restriction enzyme analysis. The method has proved to be very sensitive, with a detection limit of 20 fg of TBEV RNA per PCR run, or a single positive tick. Based on biostatistical considerations a sample size of at least 1000 ticks per estimation point was chosen. The estimated TBEV prevalence and confidence intervals (CI) were calculated from the nRT-PCR results of pooled samples (10 adults or 20 nymphs) using appropriate formulae for pooled testing. In order to identify the estimated TBEV prevalence as well as to assess the influence of annual and seasonal factors on TBEV prevalence, ticks were sampled twice a year (May and September) in 1997 and 1998 at exactly identical sites. These sites were selected because they were known to have had the highest incidence of autochthonous TBE cases during the previous 10 years. On sampling days, relevant local meteorological data were also noted. In total, 8500 I. ricinus ticks were investigated in this study, 4270 (3540 nymphs, 730 adults) from the Black Forest habitats, and 4230 (3680 nymphs, 550 adults) from the Bavarian locations. In the foci near Freiburg (Black Forest), the estimated virus prevalence was relatively high in the whole tick population, during 1997 with only slight seasonal differences [3.4% (confidence interval, CI, 2.3-4.8%) in May and 2.9% (CI 1.7-4.5%) in September]. In contrast, in 1998, in the same foci the estimated TBEV prevalence was considerably lower [1.1% (CI 0.5-2.0%) in May and 0.6% (CI 0.2-1.4%) in September]. Thus, while the seasonal differences again remained low, the annual variation was marked. In the Bavarian foci in 1997, the estimated virus prevalence of the whole tick population studied was lower than in the Black Forest foci and the seasonal fluctuations were low: in May 1997 0.9% (CI 0.4-1.8%) of the ticks were positive, in September 1.1% (CI 0.5-1.9%). In 1998, in May 2.0% (CI 1.1-3.3%) of the ticks were positive, and in September 1.1% (CI 0.5-2.1%). For the whole study period, every 50th to 100th I. ricinus nymph or adult in the Passau region was calculated to give a positive signal in the nRT-PCR. The TBEV prevalence data indicate that residents and visitors of areas in Germany known for high endemic activity take a significant risk of contracting TBEV infection, if bitten by ticks. In addition, the data suggest that annual fluctuations may exist in the whole tick population studied. Seasonal fluctuations of the virus prevalence in ticks were small.

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Role of small mammals in the ecology of Borrelia burgdorferi in a peri-urban park in north coastal California.

The role of small mammals other than woodrats in the enzootiology of the Lyme disease spirochete, Borrelia burgorferi, was assessed in the peri-urban park. Mammals were collected monthly from September through to April. Following tick removal, the animals were tested for B. burgdorferi by culture of ear-punch biopsies. Larvae and nymphs that were intermediate in morphology between Ixodes spinipalpis and Ixodes neotomae occurred on several species of rodents (Peromyscus truei, Peromyscus californicus, Microtus californicus, Rattus rattus and Reithrodontomys megalotis) and the brush rabbit (Sylvilagus bachmani). Morphometric analyses of these I. spinipalpis-like ticks and the offspring from two I. neotomae females from the site suggest that I. neotomae may bo conspecific with I. spinipalpis. Borrelia burgdorferi was isolated from eight out of 109 (7.3%), three out of 16 (18.8%), two out of 38 (5.3%) and two out of six (33.3%) P. truei, P. maniculatus, M. californicus and R. rattus, respectively. One bush rabbit yielded the first isolate of B. burgdorferi from a lagomorph in western North America. This isolate and three others derived from unfed I. spinipalpis-like nymphs failed to produce infection when inoculated intradermally into 11-12 P. maniculatus each. Likewise, no spirochetes were detected in 420 Ixodes pacificus nymphs derived from larvae fed on animals inoculated with these isolates. An additional isolate, derived from an I. spinipalpis-like nymph, was recovered by ear-punch biopsies from five our of 12 (42%) needle-inoculated P. maniculatus. However, spirochetes were not detected in 20 I. pacificus nymphs fed as larvae on each of five mice (two infected and three uninfected) inoculated with this isolate. We conclude that brush rabbits and several species of rodents besides woodrats may contribute to the maintenance of B. burgdorferi because they harbour the spirochete and are fed upon by competent enzootic vectors.

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Aspects of potato leafhopper (Homoptera: Cicadellidae) biology on Solanum berthaultii and other potato genotypes.

Survival of nymphs and adult males and females of potato leafhopper, Empoasca fabae (Harris), on six potato (Solanum spp.) genotypes with varied levels of resistance was evaluated and related to type, density, and droplet/head size of foliar glandular trichomes. Survival was greater on Solanum tuberosum cultivars ('Allegany' and 'Elba') compared with accessions of the wild S. berthaultii (PI 473331 and PI 473334). The barrier provided by glandular trichomes may affect survival of E.fabae by restricting nymph and adult feeding. PI 473331 was the most pubescent host, bearing type A and type B trichomes and the most unsuitable host for E. fabae. Allegany possesses leaflets with low trichome densities and small gland diameter and was the most suitable host for adult E. fabae survival. Females generally had higher survival rates than males. Nymphal and adult leafhoppers differed greatly in their survival rates, particularly on S. tuberosum x S. berthaultii hybrids (Q174-2 and NY123). Interestingly, nymphs had longer (190 d) mean lethal time (LT50) than adults (31 d) when held on Q174-2, and a lower LT50 (19 d) when held on NY123 compared with adults (104 d). Starvation, dehydration, or both were likely associated with mortality of nymphs confined on PI 473331 and PI 473334. Nymphs on these accessions died at a comparable rate to those confined on a starvation treatment and more rapidly than those supplied with water only, sucrose solution or other potato genotypes. Darkened trichome exudates were observed on insects held on PI 473331 and PI 473334, consistent with previous reports that glandular trichomes of S. berthaultii contain phenolic oxidation chemistry. This study aids in the identification of the probable mechanisms of resistance to E. fabae used by S. berthaulti and its hybrids with S. tuberosum.

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Glandular trichomes of Solanum berthaultii and its hybrids with Solanum tuberosum affect nymphal emergence, development, and survival of Empoasca fabae (Homoptera: Cicadellidae).

The influence of foliar glandular trichomes on developmental biology of the potato leafhopper, Empoasca fabae (Harris) (Homoptera: Cicadellidae), was examined by selective removal of trichomes on resistant and susceptible potato germplasm comprising two glandular trichome-bearing genotypes (PI 473331 and PI 473334) of wild potato, Solanum berthaultii (Hawkes), two interspecific hybrids of S. berthaultii with Solanum tuberosum L. (Q174-2 and NY123), and two commercial S. tuberosum cultivars ('Elba' and 'Allegany'). Adult mortality was affected, whereas nymphal emergence was unaffected by removal of glandular trichome exudates. No nymphs were observed emerging from leaves of PI 473331 or PI 473334. Regardless of the trichome removal treatment, no nymphs completed development on PI 473331, and only a small percentage of nymphs survived to adulthood on NY123 and PI 473334. PI 473331 had the greatest frequency of adults and nymphs with trichome exudate deposition on their bodies, followed by PI 473334. Nymphs, but not adults, had exudate accumulation when caged on foliage of Q174-2. Removal of type A trichome heads and type B droplets eliminated exudate deposition. The results of these studies provide evidence that phenolic oxidation chemistry and the physical barrier imposed by trichomes of S. berthaultii are responsible for much of the observed resistance but that other factors possibly linked to the presence of trichomes aid in the expression on resistance.

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Evaluation of Beauveria bassiana (hyphomycetes) strains as potential agents for control of Triatoma infestans (Hemiptera: Reduviidae).

Chagas disease constitutes a major human health problem in most Latin American countries. This endemic disease is transmitted by several species of triatomine bugs, the most important being Triatoma infestans (Klug). In this article, we report on the selection of strains of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuill. virulent to T. infestans for possible use as bioinsecticides. Four strains of B. hassiana isolated from Argentina (Bb 1, 10, 25, and 65) were evaluated. To calculate mortality and mean lethal time, nymphs and adults of T. infestans were treated with conidia produced on complete agar medium and wheat brain and rice husk medium (WH). The LD50 for nymphs and adults of T. infestans was calculated. The effect of different temperatures (18, 22, 26, 30, and 34 degrees C) and relative humidities (35 and 90% RH) on mortality of nymphs were studied. We evaluated the compatibility of strains with Deltamethrin and Beta-Cypermethrin. Although the strain Bb 25 failed to grow on WH, the other three strains showed similar mortality values independent of the culture medium used to grow the microorganisms. The lowest LD50 values for nymphs were obtained with the strains Bb 10 and 65 and for adults were Bb 1, 10, and 65. The highest mean mortality was obtained with strain Bb 10, and among temperatures the highest mean mortality was observed at 26 degrees C. Relative humidity did not affect the mortality of T. infestans nymphs with all strains and temperatures assayed. Deltamethrin did not affect any of the three strains for the four studied doses, and Beta-Cypermethrin could be used in combination with the fungus only at low doses. The strain Bb 10 was selected for future assays under natural climatic conditions.

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Refractoriness of the western fence lizard (Sceloporus occidentalis) to the Lyme disease group spirochete Borrelia bissettii.

The western fence lizard, Sceloporus occidentalis, is refractory to experimental infection with Borrelia burgdorferi sensu stricto, one of several Lyme disease spirochetes pathogenic for humans. Another member of the Lyme disease spirochete complex, Borrelia bissettii, is distributed widely throughout North America and a similar, if not identical, spirochete has been implicated as a human pathogen in southern Europe. To determine the susceptibility of S. occidentalis to B. bissettii, 6 naïve lizards were exposed to the feeding activities of Ixodes pacificus nymphs experimentally infected with this spirochete. None of the lizards developed spirochetemias detectable by polymerase chain reaction for up to 8 wk post-tick feeding, infected nymphs apparently lost their B. bissettii infections within 1-2 wk after engorgement, and xenodiagnostic L. pacificus larvae that co-fed alongside infected nymphs did not acquire and maintain spirochetes. In contrast, 3 of 4 naïve deer mice (Peromyscus maniculatus) exposed similarly to feeding by 1 or more B. bissettii-infected nymphs developed patent infections within 4 wk. These and previous findings suggest that the complement system of S. occidentalis typically destroys B. burgdorferi sensu lato spirochetes present in tissues of attached and feeding I. pacificus nymphs, thereby potentially reducing the probability of transmission of these bacteria to humans or other animals by the resultant adult ticks.

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Effect of sedimentary cadmium on the behavior of a burrowing mayfly (Ephemeroptera, Hexagenia limbata).

Behavioral responses of aquatic invertebrates can serve as precocious indicators of adverse effects caused by metals. We conducted experiments to determine if the behavior of nymphs of the burrowing mayfly Hexagenia limbara (Ephemeroptera) was modified by the presence of cadmium (Cd) in sediments. We first exposed H. limbata to concentrations of sedimentary Cd found in lakes subject to smelter emissions (20 microg/g dry wt). We then compared the burrowing activity of control and Cd-contaminated nymphs by means of radiographs. We also studied the time allotted by nymphs to various activities (irrigation, walking, burrowing/feeding, cleaning, and turning) as determined from films taken using an infrared video camera. The results obtained suggest no significant difference in the burrowing activity of control versus Cd-contaminated individuals. Furthermore, the time spent in various behavioral activities was similar between control and Cd-contaminated nymphs. Given these results, we conclude that Cd has no effect on the behavior of H. limbata for the range of Cd concentrations that we measured in nymphs.

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Transmission of Crimean-Congo hemorrhagic fever virus in two species of Hyalomma ticks from infected adults to cofeeding immature forms.

Crimean-Congo hemorrhagic fever (CCHF) virus was transmitted from infected adult Hyalomma ticks to uninfected larval and nymphal Hyalomma ticks while cofeeding on a guinea pig host that did not have a detectable viremia. When tested after feeding with infected adults, three (0.8%) of 370 H. truncatum larvae contained detectable CCHF virus (mean virus titer 10(1.6) plaque-forming units [PFU]/tick). The virus was transmitted transstadially from infected larvae and was detected in 15 (1.2%) of 1,253 nymphs and 12 (0.1%) of 2,049 adults. Virus was recovered from 18 (1.9%) of 931 H. impeltatum nymphs, which originated from larvae that cofed with infected adults. After H. impeltatum nymphs cofed with infected adults, CCHF virus was detected in 21 (4.3%) of 449 (mean virus titer 10(1.7) PFU/tick) fed nymphs, but none of 886 adults tested after molt. Results of this study indicate that a small proportion of either larvae or nymphs may acquire CCHF infection while cofeeding on a host without a detectable viremia.

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