Selection for fenpropathrin and fenpropathrin + acephate resistance in the silverleaf whitefly (Homoptera: Aleyrodidae)

A large, genetically diverse pool of Bemisia argentifolii Bellows & Perring was collected in 1994 from different crops, and a mixed colony was established in the laboratory. Subsets of this colony were reared on cotton plants held in large Plexiglas cages, and adult whiteflies were selected for resistance to fenpropathrin and to fenpropathrin + acephate (1:5). Selection was performed by exposing adults to treated glass vials at doses sufficient to give 60-80% mortality. Thirteen generations of adult selection with fenpropathrin + acephate yielded 856.3- and 1,289.3-fold tolerance (using lethal concentration ratio), respectively, to fenpropathrin and to fenpropathrin + acephate, indicating additive genetic variation for resistance in the source population. Selection with fenpropathrin alone yielded only a 10.9-fold increase in tolerance to fenpropathrin at the end of the selection period. There was no significant change in tolerance to fenpropathrin + acephate in this strain. Contrary to expectations, the early onset and the magnitude of resistance attained on selection with fenpropathrin + acephate compared with fenpropathrin alone indicates that this mixture evidently possesses a high degree of selectivity for development of resistance in B. argentifolii. Estimates of realized heritability of resistance to fenpropathrin and to fenpropathrin + acephate in B. argentifolii (in the fenpropathrin + acephate-selected strain) showed that they were significantly higher in the first half of selection (six generations), in both instances. Rearing of the fenpropathrin + acephate-resistant strain under conditions free of insecticides for six generations did not result in any significant decline in resistance, indicating that resistance is fairly stable.     

Increase in cold-shock tolerance by selection of cold resistant lines in Drosophila melanogaster

Abstract.

• 1 In Drosophila melanogaster, the cold-shock tolerance of adult flies at -7°C increased 22% after a prior 2h exposure to 4°C as measured by LD₅₀, the dose (degree minutes of exposure to subzero temperature) which resulted in 50% mortality.
• 2 Cold-shock tolerance was further significantly increased by selecting cold resistant lines by exposure of adults (1) to 4°C for 2 h (short-term chilling), or (2) to -7°C for 80–120 min (cold shock), or (3) to short-term chilling followed by cold-shock.
• 3 After ten generations of selection, the greatest increase in cold-shock tolerance was found in flies selected using the combined exposure of short-term chilling and cold shock. LD₅₀s increased 33% in comparison with the unselected control strain when no chilling pre-treatment was given prior to cold shock at -7°C.
• 4 The rapid cold-hardening response increased 82% in the line selected by the short-term chilling and cold-shock regime.
• 5 The enhanced cold-shock tolerance was relatively stable since no decrease was observed after four generations without selection.
• 6 This report shows the role of short-term adaptation as well as selection in the capacity to survive low temperatures in non-diapausing stages of insects.

     

Comparative proteomics reveals mechanisms that underlie insecticide resistance in Culex pipiens pallens Coquillett

Mosquito control based on chemical insecticides is considered as an important element of the current global strategies for the control of mosquito-borne diseases. Unfortunately, the development of insecticide resistance of important vector mosquito species jeopardizes the effectiveness of insecticide-based mosquito control. In contrast to target site resistance, other mechanisms are far from being fully understood. Global protein profiles among cypermethrin-resistant, propoxur-resistant, dimethyl-dichloro-vinyl-phosphate-resistant and susceptible strain of Culex pipiens pallens were obtained and proteomic differences were evaluated by using isobaric tags for relative and absolute quantification labeling coupled with liquid chromatography/tandem mass spectrometric analysis. A susceptible strain of Culex pipiens pallens showed elevated resistance levels after 25 generations of insecticide selection, through iTRAQ data analysis detected 2,502 proteins, of which 1,513 were differentially expressed in insecticide-selected strains compared to the susceptible strain. Finally, midgut differential protein expression profiles were analyzed, and 62 proteins were selected for verification of differential expression using iTRAQ and parallel reaction monitoring strategy, respectively. iTRAQ profiles of adaptation selection to three insecticide strains combined with midgut profiles revealed that multiple insecticide resistance mechanisms operate simultaneously in resistant insects of Culex pipiens pallens. Significant molecular resources were developed for Culex pipiens pallens, potential candidates were involved in metabolic resistance and reducing penetration or sequestering insecticide. Future research that is targeted towards RNA interference of the identified metabolic targets, such as cuticular proteins, cytochrome P450s, glutathione S-transferases and ribosomal proteins proteins and biological pathways (drug metabolism—cytochrome P450, metabolism of xenobiotics by cytochrome P450, oxidative phosphorylation, ribosome) could lay the foundation for a better understanding of the genetic basis of insecticide resistance in Culex pipiens pallens.     

A strain of Bacillus sphaericus causes slower development of resistance in Culex quinquefasciatus

Two field-collected Culex quinquefasciatus colonies were subjected to selection pressure by three strains of Bacillus sphaericus, C3-41, 2362, and IAB59, under laboratory conditions. After 13 and 18 generations of exposure to high concentrations of C3-41 and IAB59, a field-collected low-level-resistant colony developed >144,000- and 46.3-fold resistance to strains C3-41 and IAB59, respectively. A field-collected susceptible colony was selected with 2362 and IAB59 for 46 and 12 generations and attained >162,000- and 5.7-fold resistance to the two agents, respectively. The pattern of resistance evolution in mosquitoes depended on continuous selection pressure, and the stronger the selection pressure, the more quickly resistance developed. The resistant colonies obtained after selection with B. sphaericus C3-41 and 2362 showed very high levels of cross-resistance to B. sphaericus 2362 and C3-41, respectively, but they displayed only low-level cross-resistance to IAB59. On the other hand, the IAB59-selected colonies had high cross-resistance to both strains C3-41 and 2362. Additionally, the slower evolution of resistance against strain IAB59 may be explained by the presence of another larvicidal factor. This is in agreement with the nontoxicity of the cloned and purified binary toxin (Bin1) of IAB59 for 2362-resistant larvae. We also verified that all the B. sphaericus-selected colonies showed no cross-resistance to Bacillus thuringiensis subsp. israelensis, suggesting that it would be a promising alternative in managing resistance to B. sphaericus in C. quinquefasciatus larvae.     

Clostridium bifermentans serovar malaysia, a new anaerobic bacterium pathogen to mosquito and blackfly larvae

A strain of Clostridium bifermentans individualized as serovar malaysia (C.b.m.) according to its specific H antigen is toxic to mosquito and blackfly larvae when given orally. The toxicity occurs in sporulated cells which contain, in addition to spores, proteinic parasporal inclusion bodies and feather-like appendages; the amino acid content of the inclusion bodies is similar to that of Bacillus thuringiensis serovar israelensis (B.t.i.) and B. sphaericus crystals. The toxicity to Anopheles stephensi is as high as that of B.t.i. and the best strains of B. sphaericus. Culex pipiens is somewhat less susceptible, and Aedes aegypti much less. Pure parasporal inclusion bodies, isolated by ultracentrifugation on sucrose gradients, are highly toxic to mosquito larvae. The larvicidal power is destroyed by heating at 80 degrees C or by treatment with 50 mM NaOH. It is preserved by freeze-drying. The innocuity to mice of the sporulated cells is shown by different routes of administration: force-feeding, percutaneous, subcutaneous, intraperitoneal or intravenous injections. The potential for the biological control of mosquito and blackfly larvae is suggested.     

Cross-resistance between strains of Bacillus sphaericus but not B. thuringiensis israelensis in colonies of the mosquito Culex quinquefasciatus

Two colonies of Culex quinquefasciatus Say (Diptera: Culicidae) were selected with Bacillus sphaericus strains C3-41 and IAB59 in the laboratory for 13 and 18 generations; they attained 145,000- and 48.3-fold resistance, respectively, in comparison with a susceptible laboratory colony (SLCq) and showed very high levels of cross-resistance (8500- to 145,000-fold) to B. sphaericus strains C3-41, 1593, 2297 and 2362. They were relatively susceptible to B. sphaericus strains LP1-G and 47-6B (only 0.8- to 2.8-fold tolerance), with 24.8- to 48.3-fold cross-resistance to strain IAB59. B. sphaericus-resistant mosquito colonies remained highly susceptible to B. thuringiensis israelensis, suggesting that B.t.i. would be of value in the management of B. sphaericus-resistant Cx. quinquefasciatus colonies. The demonstration of low or no cross-resistance of two selected resistant Cx. quinquefasciatus colonies to IAB59, LP1-G and 47-6B strains of B. sphaericus and the finding of a major 49 kDa protein in these strains suggest that there is likely to be another mosquitocidal factor in the three strains.     

ECOLOGICAL NOVELTY BY HYBRIDIZATION: EXPERIMENTAL EVIDENCE FOR INCREASED THERMAL TOLERANCE BY TRANSGRESSIVE SEGREGATION IN TIGRIOPUS CALIFORNICUS

Early generations of hybrids can express both genetic incompatibilities and phenotypic novelty. Insights into whether these conflicting interactions between intrinsic and extrinsic selection persist after a few generations of recombination require experimental studies. To address this question, we use interpopulation crosses and recombinant inbred lines (RILs) of the copepod Tigriopus californicus, and focus on two traits that are relevant for the diversification of this species: survivorship during development and tolerance to thermal stress. Experimental crosses between two population pairs show that most RILs between two heat‐tolerant populations show enhanced tolerance to temperatures that are lethal to the respective parentals, whereas RILs between a heat‐tolerant and a heat‐sensitive population are intermediate. Although interpopulation crosses are affected by intrinsic selection at early generational hybrids, most of the sampled F₉ RILs have recovered fitness to the level of their parentals. Together, these results suggest that a few generations of recombination allows for an independent segregation of the genes underlying thermal tolerance and cytonuclear incompatibilities, permitting certain recombinant lineages to survive in niches previously unused by parental taxa (i.e., warmer thermal environments) without incurring intrinsic selection.     

Does selection on increased cold tolerance in the adult stage confer resistance throughout development?

Artificial selection is a powerful approach to unravel constraints on genetic adaptation. Although it has been frequently used to reveal genetic trade-offs among different fitness-related traits, only a few studies have targeted genetic correlations across developmental stages. Here, we test whether selection on increased cold tolerance in the adult stage increases cold resistance throughout ontogeny in the butterfly Bicyclus anynana. We used lines selected for decreased chill-coma recovery time and corresponding controls, which had originally been set up from three levels of inbreeding (outbred control, one or two full-sib matings). Four generations after having terminated selection, a response to selection was found in 1-day-old butterflies (the age at which selection took place). Older adults showed a very similar although weaker response. Nevertheless, cold resistance did not increase in either egg, larval or pupal stage in the selection lines but was even lower compared to control lines for eggs and young larvae. These findings suggest a cost of increased adult cold tolerance, presumably reducing resource availability for offspring provisioning and thereby stress tolerance during development, which may substantially affect evolutionary trajectories.     

用刘氏中剑水蚤防治蚊幼虫(英文)

本研究在室内评价了在济宁采集的捕食性刘氏中剑水蚤单用及与球形芽孢杆菌(B.s.)或苏云金杆菌以色列变种(B.t.i.)合用对蚊幼虫的控制效果。在室内,刘氏中剑水蚤对白蚊伊蚊Ⅰ龄幼虫的捕食效果好于对中华按蚊和淡色库蚊幼虫的效果。蚊笼模拟试验表明,刘氏中剑水蚤可分别于9、10和7周内消灭淡色库蚊、中华按蚊和白蚊伊蚊的幼虫种群。该剑水蚤增长速率快,大量培养经济易行。现场试验证明,刘氏中剑水蚤虽可在蚊虫孳生地存活时间长于蚊虫季节,但单用该剑水蚤不能象化学杀虫剂一样快速完全地控制三种蚊虫。刘氏中剑水蚤与B.s.或B.t.i.合用,应用刘氏中剑水蚤的长持效和生物杀虫剂的速效,取得了高效、持久的防治效果。因此二者合用可用于蚊虫防治并可作为蚊虫综合防治的一个组成部分。     

BIOLOGICAL CONTROL OF MOSQUITO LARVAE WITH MESOCYCLOPS LEUKARTI1)

Abstract  The control efficacy of a predatory copepod, Mesocyclops leukarti , collected in Jining, and its compatibility with Bacillus sphaericus (B.s. ) and B. thuringiensis var. csraefensis H-14 ( B. t. i. ) against mosquito larvae were evaluated in the laboratory and in the field. In the laboratory, M. leukarti showed higher predatory activity toward first instar larvae of Aedes albopictus than toward that of Anopheles sinensis and Culex pipiens pallens. Cage simulation test showed that larvae of Cx. pipiens pallens, An. sinensis and Ae. albopictus could be eliminated within 9, 10 and 7 weeks after interaction with the copepods respectively. M. leukarti population can grow quickly, and its mass production is easy and economic. However, pilot field trials indicated that M. leukarti alone could not eradicate mosquito larvae of the three species rapidly and completely as by chemical insecticides, although they could persist in mosquito breeding habitats longer than the mosquito season. When Mesocyclops was used in combination with B. s. and B. t. i. , higher and sustainable control effects were achieved. So this combination, which achieved the rapid larvicidal and sustainable control effect, was presumed to be practical and could be integrated into mosquito control program.     

Ethanol tolerances of Drosophila melanogaster populations selected on different concentrations of ethanol supplemented media

Summary Eight recently collected Australasian populations of D. melanogaster were each divided into eight selection lines. Two of these lines from each population were maintained on one of four types of selection media: standard food supplemented with 0%, 3%, 6% and 9% ethanol. After 30 generations the selection lines were tested for tolerance to 9% ethanol medium and after another 20 generations adults were tested for tolerance to concentrated ethanol fumes. Significant differences in tolerance were found among lines selected on different media which were consistent across the eight populations. On the 9% test media, the 6% and 9% selection lines, as compared with the control lines selected on 0% ethanol, were more likely to survive as pre-adults or adults, faster to develop as preadults, and heavier and more productive as adults. However, the tolerance of the 3% lines to the 9% test media was less than that of the 0% control lines in preadult and adult survival, intermediate between that of the 0% and the 6% and 9% lines in productivities, and apparently superior to the 6% and 9% lines in development times and adult weights. The 3%, 6% and 9% lines showed similar tolerances to the ethanol vapour. Previous work showed that 3% ethanol can be a metabolic benefit to D. melanogaster but 6% and 9% are metabolic costs. The present results suggest that the phenotype selected on 3% to obtain a metabolic benefit differs in many respects from that selected on 6% and 9% to minimise their detrimental effects.     

Ticks and mosquitoes as vectors of Borrelia burgdorferi s. l. in the forested areas of Szczecin

The aim of the study was to determine the infection level of adult forms and larvae of ticks and mosquitoes with Borrelia burgdorferi in the forested areas of Szczecin. A total of 1699 ticks Ixodes ricinus, including 1422 nymphs, 277 adult forms and 2862 mosquito females representing the genera Aedes (89.6%) and Culex (10.4%) were collected between the years 2004 and 2005. A further 3746 larvae and 1596 pupae of Culex pipiens pipiens were colleted from water bodies. Borrelia burgdorferi s. l. was detected in the arthropods by the method of indirect immunofluorescence assay (IFA). A positive immunological reaction was detected in 16.6% of the adult forms and in 16.5% of the nymphs of Ixodes ricinus. Spirochetes were also detected in 1.7% of mosquito females, 3.2% of larvae and in 1.6% of pupae of Culex pipiens pipiens. The results of the present study confirm that contact with ticks constitutes the main risk of contracting Lyme disease, although mosquitoes play a role as vectors as well.     

Distribution and reproductive strategies of Gyrinicola batrachiensis (Oxyuroidea: Pharyngodonidae) in larvae of eight species of amphibians from Nebraska

In total, 462 tadpoles and salamander larvae of 8 species were examined for the presence of Gyrinicola batrachiensis from 5 locations in Nebraska. Infection by G. batrachiensis occurred in tadpoles of Rana blairi , Rana catesbeiana, Rana pipiens, and Bufo woodhousii. Tadpoles of Hyla chrysoscelis , Spea bombifrons, and Pseudacris maculata and larvae of Ambystoma mavortium were not infected with G. batrachiensis. Population structure, defined as prevalence, mean abundance, and mean intensity of G. batrachiensis, varied among tadpoles of different amphibian species and was determined by collection locality, developmental period of tadpole hosts, amphibian species co-occurrence, and different reproductive strategies of G. batrachiensis , or a combination. Gyrinicola batrachiensis observed in all ranid tadpoles and B. woodhousii tadpoles from where bufonids were the only anuran species present, confirmed to the didelphic haplodiploidy and monodelphic parthenogenetic reproductive strategies, respectively. However, tadpoles of B. woodhousii that co-occurred with tadpoles of R. pipiens at Cedar Creek were inconsistent with these predictions and contained both male and didelphic female nematodes, but at a low mean intensity (1.61 ± 0.70). Didelphic female nematodes from B. woodhousii tadpoles at Cedar Creek only produced thick-shelled eggs, whereas nematodes in R. pipiens tadpoles had a high mean intensity (14.88 ± 23.83) from this location and contained both thick-shelled and thin-shelled eggs in their respective uteri. More importantly, adult female nematodes from tadpoles of R. pipiens and B. woodhousii from Cedar Creek were morphologically more similar to each other than to female nematodes recovered from tadpoles of other anuran species, other locations, or both. These data suggest that when strains of G. batrachiensis are shared by tadpoles of different amphibian species that differ in developmental period, the nematodes have an intermediate reproductive strategy in amphibian species, with tadpoles having short development.     

QTL mapping of terminal heat tolerance in hexaploid wheat (T. aestivum L.)

High temperature (>30 °C) at the time of grain filling is one of the major causes of yield reduction in wheat in many parts of the world, especially in tropical countries. To identify quantitative trait loci (QTL) for heat tolerance under terminal heat stress, a set of 148 recombinant inbred lines was developed by crossing a heat-tolerant hexaploid wheat (Triticum aestivum L.) cultivar (NW1014) and a heat-susceptible (HUW468) cultivar. The F(5), F(6), and F(7) generations were evaluated in two different sowing dates under field conditions for 2 years. Using the trait values from controlled and stressed trials, four different traits (1) heat susceptibility index (HSI) of thousand grain weight (HSITGW); (2) HSI of grain fill duration (HSIGFD); (3) HSI of grain yield (HSIYLD); and (4) canopy temperature depression (CTD) were used to determine heat tolerance. Days to maturity was also investigated. A linkage map comprising 160 simple sequence repeat markers was prepared covering the whole genome of wheat. Using composite interval mapping, significant genomic regions on 2B, 7B and 7D were found to be associated with heat tolerance. Of these, two (2B and 7B) were co-localized QTL and explained more than 15 % phenotypic variation for HSITGW, HSIGFD and CTD. In pooled analysis over three trials, QTL explained phenotypic variation ranging from 9.78 to 20.34 %. No QTL × trial interaction was detected for the identified QTL. The three major QTL obtained can be used in marker-assisted selection for heat stress in wheat.     

Assessment of fitness costs of resistance against the parasitoid Leptopilina victoriae in Drosophila bipectinata

How insects evolve resistance or counter-resistance against antagonists is a basic issue in the study of host-parasitoid coevolution. One of the factors that affect their coevolution is fitness costs of resistance and counter-resistance. Here, we assess fitness costs of resistance against the parasitoid Leptopilina victoriae in Drosophila bipectinata on the basis of selection experiments. We made a base population by mixing three geographic fly populations that differed in resistance. The established base population was divided into four populations, two for selection of resistance against a L. victoriae population and two for control. Resistance increased rapidly in response to selection and reached a very high level within four generations in the selected populations, while resistance of the control populations remained low during 20 generations. High resistance of the selected populations was maintained at least for 10 generations after selection was stopped. Both selected populations had lower female longevity than the control populations, and at least one of the selected populations had shorter thorax length, lower female desiccation tolerance and adult heat tolerance than both or either of the control populations. On the other hand, the selected populations had higher male starvation tolerance and longevity than the control populations. There were no significant differences in resistance against another population of L. victoriae and two other parasitoid species between the selected and control populations. These results suggest that the resistance against the L. victoriae population in D. bipectinata may incur some but not so high costs and act parasitoid-species- and/or parasitoid-population-specifically.     

Indirect selection for increased susceptibility to permethrin in diamondback moth (Lepidoptera: Plutellidae)

We have been exploring the behavioral response ot insect pests to heterogeneous distribution of toxins (low dose with refugia), and its genetic correlation with physiological tolerance to these toxins. A field-collected population of diamondback moth, Plutella xylostellu (L.) (Lepidoptera Plutellidae), from Celeryville, OH, was selected with permethrin to determine whether low heterogeneous doses could lead to increased susceptibility to permethrin by selecting indirectly on behavior. Two replicates of each of three selection regimes: uniform high concentration hypothesized to result in increased physiological tolerance, heterogeneous low concentration hypothesized to result in increased susceptibility through indirect selection on behavior, and a control with no exposure to permethrin, were maintained in 1-in3 cages in a greenhouse, for 33 generations. All life stages of the diamondback moth were exposed to the selection regimes, and new generations were started with a random selection of pupae from the previous generation. Lines selected with uniform high concentrations developed 76-fold levels of resistance to permethrin by the 17th generation, with little changes thereafter. For generations 1-20, lines selected with heterogeneous low concentrations remained slightly lower in LC50 but not significantly different from the unselected control lines. Based on confidence intervals from probit analyses, the LC50 of the lines selected with heterogeneous low concentration, however, were significantly lower than those of the control lines in generations 21-33. Our results demonstrate that selection on behavioral responses can result in greater susceptibility than no selection at all, despite exposure to the toxin and ample genetic variation and potential for increased physiological tolerance. The implications of our findings, which are based on selection scenarios that could take place in field situations, are that behavioral responses can prevent and even decrease the levels of resistance in insect populations, an important result with respect to resistance and resistance management.     

Cyt1A of Bacillus thuringiensis delays evolution of resistance to Cry11A in the mosquito Culex quinquefasciatus

Insecticides based on Bacillus thuringiensis subsp. israelensis have been used for mosquito and blackfly control for more than 20 years, yet no resistance to this bacterium has been reported. Moreover, in contrast to B. thuringiensis subspecies toxic to coleopteran or lepidopteran larvae, only low levels of resistance to B. thuringiensis subsp. israelensis have been obtained in laboratory experiments where mosquito larvae were placed under heavy selection pressure for more than 30 generations. Selection of Culex quinquefasciatus with mutants of B. thuringiensis subsp. israelensis that contained different combinations of its Cry proteins and Cyt1Aa suggested that the latter protein delayed resistance. This hypothesis, however, has not been tested experimentally. Here we report experiments in which separate C. quinquefasciatus populations were selected for 20 generations to recombinant strains of B. thuringiensis that produced either Cyt1Aa, Cry11Aa, or a 1:3 mixture of these strains. At the end of selection, the resistance ratio was 1,237 in the Cry11Aa-selected population and 242 in the Cyt1Aa-selected population. The resistance ratio, however, was only 8 in the population selected with the 1:3 ratio of Cyt1Aa and Cry11Aa strains. When the resistant mosquito strain developed by selection to the Cyt1Aa-Cry11Aa combination was assayed against Cry11Aa after 48 generations, resistance to this protein was 9.3-fold. This indicates that in the presence of Cyt1Aa, resistance to Cry11Aa evolved, but at a much lower rate than when Cyt1Aa was absent. These results indicate that Cyt1Aa is the principal factor responsible for delaying the evolution and expression of resistance to mosquitocidal Cry proteins.     

Wolbachia in the Culex pipiens group mosquitoes: introgression and superinfection

Wolbachia bacteria in mosquitoes induce cytoplasmic incompatibility (CI), where sperm from Wolbachia-infected males can produce inviable progeny. The wPip strain in the Culex pipiens group of mosquitoes produces a complexity of CI crossing types. Several factors are thought to be capable of influencing the expression of CI including Wolbachia strain type and host genotype. In this study, the unidirectional CI that occurs between 2 C. pipiens complex laboratory strains, Col and Mol, was further investigated by nuclear genotype introgression. The unidirectional CI between Col and Mol was not found to be influenced by host genetic background, in contrast to a previous introgression study carried out using bidirectionally incompatible C. pipiens group strains. A line containing both wPip strain variants superinfection was also generated by embryonic cytoplasmic transfer. The same crossing type as the parental Col strain was observed in the superinfected line. Quantitative polymerase chain reaction demonstrated a low density of the injected wPipMol variant in the superinfected line after 18 generations, which was considered likely to be responsible for the crossing patterns observed. The Wolbachia density was also shown to be lower in the parental Mol strain males compared with Col strain males, and no inverse relationship between WO phage and Wolbachia density could be detected.     

SELECTION FOR HEAT-SHOCK RESISTANCE IN LARVAL AND IN ADULT DROSOPHILA BUZZATII: COMPARING DIRECT AND INDIRECT RESPONSES

Direct and correlated responses in selection for heat-shock resistance in adult and in larval Drosophila buzzatii were studied. Two lines were artificially selected for higher survival to heat stress as adults, and two other lines were reared under a fluctuating thermal environment as larvae, 35°C for 6 h and 25°C for 18 h, to “naturally” select for higher resistance as larvae. The latter two lines were duplicated after nine generations to yield additional lines to be “naturally” selected as larvae at a higher temperature, 38.2°C for 6 h. Control lines were maintained separately for the adult and larval selection lines. A significant direct response to selection was found for the adult selection lines. However, larvae of these adult selection lines were no more heat resistant than were larvae of the control lines. One of the two larval selection lines increased significantly in heat resistance as larvae. However, adult heat resistance was similar for lines selected as larvae and the corresponding control lines maintained at 25°C. Changes in developmental time accompanied changes in survival after stress in both sets of lines selected for increased heat resistance.     

T regulatory cells contribute to the attenuated primary CD8+ and CD4+ T cell responses to herpes simplex virus type 2 in neonatal mice

The first weeks of life are characterized by immune tolerance and increased susceptibility to intracellular pathogens. The neonatal adaptive response to HSV is attenuated compared with adult control models in humans and mice. T Regulatory cells (Tregs) control autoimmunity and excessive immune responses to infection. We therefore compared Treg responses in the draining lymph nodes (LN) of HSV-infected neonatal and adult C57BL/6 mice with the effect of Treg depletion/inactivation by anti-CD25 (PC61) treatment before infection on Ag-specific T cell effector responses at this site. There was a small, but significant increase in the frequency of CD4(+)Foxp3(+) Tregs at day 3 postinfection (p.i.) in the LN of neonatal and adult mice, compared with age-matched mock-infected controls. Depletion of Tregs before HSV infection significantly enhanced HSV-specific CD8(+) T cell cytotoxicity in vivo, cell number, activation, and granzyme B expression 4 days p.i. only in neonatal mice, and significantly enhanced CD8(+) and CD4(+) T cell IFN-gamma responses in both infected adults and neonates. Treg depletion also reduced the titer of infectious virus in the draining LN and nervous system of infected neonates on days 2 and 3 p.i. Treg suppression of the neonatal CTL response p.i. with HSV was associated with increased expression of TGF-beta in the draining LN at day 4 p.i. compared with uninfected neonates, but IL-10 was increased in infected adults alone. These experiments support the notion that the newborn primary T cell effector responses to HSV are suppressed by Tregs.