Vertical transmission of the entomopathogenic soil fungus Scopulariopsis brevicaulis as a contaminant of eggs in the winter tick, Dermacentor albipictus, collected from calf moose (New Hampshire, USA).

Moose naturally acquire soil fungi on their fur that are entomopathogenic to the winter tick, Dermacentor albipictus. Presumed to provide a measure of on-host tick control, it is unknown whether these soil fungi impact subsequent off-host stages of the tick. Eggs and resultant larvae originating from engorged, adult female winter ticks collected from dead calf moose (Alces alces) were used to investigate the presence and extent of fungal infection. Approximately 40% of eggs and larvae were infected, almost exclusively by the fungus Scopulariopsis brevicaulis (teleomorph Microascus brevicaulis: Microascaceae, Ascomycota). Eggs analysed on the day of oviposition and day of hatching had high frequency (40%) of S. brevicaulis, whereas the frequency in eggs harvested in utero was minimal (7%); therefore, exposure occurs pre-oviposition in the female's genital chamber, not by transovarial transmission. At hatching, larvae emerge containing S. brevicaulis indicating transstadial transmission. Artificial infection by topical application of eggs and larvae with a large inoculum of S. brevicaulis spores caused rapid dehydration, marked mortality; pathogenicity was confirmed by Koch's postulates. The high hatching success (>90%) and multi-month survival of larvae imply that S. brevicaulis is maintained as a natural pathobiont in winter ticks.

Tick development on sexually-active bull moose is more advanced compared to that of cow moose in the winter tick, Dermacentor albipictus.

We performed a complete survey of ticks on 100 cm2 skin samples collected from 30 moose (Alces alces) harvested in 2017 in central and northern Maine, U.S.A. The samples were collected from 15 bulls, 13 cows, and 2 calves in mid-October when moose are breeding and winter ticks (Dermacentor albipictus) quest for a host. We identified only winter ticks with 99.2% in a juvenile stage; 3 adult ticks were found. Unfed nymphs were most common on bulls, whereas most ticks were fed larvae on cows and calves. The mean total count on bull samples was 21 ±â€¯4.4 (range = 0-55) and higher than on cows (6 ±â€¯0.5; range = 2-8). Unlike previous surveys, tick abundance was lowest on calves. Tick abundance was independent of age or weight of adult moose. The higher abundance and more rapid development of winter ticks on adult bulls likely reflects the seasonal influence of increased movements and hormonal cycles associated with reproduction.

Low and high thermal tolerance characteristics for unfed larvae of the winter tick Dermacentor albipictus (Acari: Ixodidae) with special reference to moose.

We report that larvae of the winter tick Dermacentor albipictus, the only stage that will quest for a host, can tolerate short-term cold shock down to -25°C and short-term heat shock as high as 46°C. Unlike a three host-tick, larvae of D. albipictus have no preconditioning response to low or high temperature exposure by rapid cold hardening (RCH) or heat hardening, and poor ability to acclimate to low and high temperature extremes. Thermal tolerance limits were not improved as the result of larval clustering, and there was only a minimal effect due to changes in photoperiod. These larvae are freeze intolerant and die at higher temperatures (-5 to -10°C) from contact with ice by inoculative freezing. In absence of cold-associated resistance mechanisms, winter survival requires that larvae procure a host before the first snow cover. Their low and high temperature tolerance, however, is a key survival element that adapts them for off-host periods during summer, which in the arctic could allow for northern expansion.

Short day-triggered quiescence promotes water conservation in the American dog tick, Dermacentor variabilis.

A short day-long day exposure comparison of water balance characteristics throughout the life history of the American dog tick, Dermacentor variabilis, was conducted to examine how this tick survives winter-associated dryness. Larvae, nymphs, and adults under potential diapause-inducing conditions (short day, 20 °C) have low water loss rates compared to long day exposed ticks. Above the critical equilibrium activity, these nonfed ticks show greater water gain from lower water vapor activities. In contrast to nonfed stages, eggs and blood-engorged stages (fed larva, fed nymph) were enhanced for water conservation independent of short day exposure and did not display reduced water loss rates under short day conditions. This short day-prompted response in nonfed ticks was distinguished from a genetically programmed diapause as noted in other arthropods by: (1) reduction in water loss was temporary and could be reversed quickly by transferring to long day for only a single day, then re-triggered by returning to short day conditions; (2) switch between low and high water loss rates could be induced multiple times and for multiple different stages; and (3) response by ticks was to the photoperiod they were under, not the condition that was experienced by previous fed stages or in the mother in the case of eggs. Furthermore, short day had no impact on altering body water content, dehydration tolerance limit, instar yields, or developmental stadia. We conclude that a short day-triggered quiescence reduces desiccation stress associated with overwintering in nonfed ticks. This effect on water conservation due to short day exposure is more pronounced for subadults and is likely due to their increased sensitivity to water loss as a result of their smaller size, which is critical to prevent dehydration during the dry winter periods.

Behavioral correction to prevent overhydration and increase survival by larvae of the net-spinning caddisflies in relation to water flow.

We report behavioral regulation of body water content in caddisfly larvae, Hydropsyche morosa and Cheumatopsyche pettiti, by selecting microhabitats with different water flow rates. The purpose of our study was to examine features necessary for survival in the same apparent habitat, because the two species co-exist in riffle areas of freshwater streams. Both species are highly sensitive to water loss as a result of high water loss rates and depend on immersion in fresh water (hypo-osmotic) to maintain water stores. In contrast to C. pettiti, H. morosa is larger, retains water more effectively, and features reduced water loss rates with suppressed activation energies. When H. morosa was confined to areas of low or no water flow, overhydration led to rapid mortality, whereas the same conditions favored water balance maintenance and survival in C. pettiti. In attraction bioassays, H. morosa moved and remained within areas of high water flow and C. pettiti preferred areas with low water flow. Because water flow rates are unlikely to directly impact water gain, the mechanism responsible for increased survival and water balance maintenance is likely related to the impact of water flow on oxygen availability, differences in feeding ecology, or other underlying factors.

Fungicide contamination reduces beneficial fungi in bee bread based on an area-wide field study in honey bee, Apis mellifera, colonies.

Fermentation by fungi converts stored pollen into bee bread that is fed to honey bee larvae, Apis mellifera, so the diversity of fungi in bee bread may be related to its food value. To explore the relationship between fungicide exposure and bee bread fungi, samples of bee bread collected from bee colonies pollinating orchards from 7 locations over 2 years were analyzed for fungicide residues and fungus composition. There were detectable levels of fungicides from regions that were sprayed before bloom. An organic orchard had the highest quantity and variety of fungicides, likely due to the presence of treated orchards within bees' flight range. Aspergillus, Penicillium, Rhizopus, and Cladosporium (beneficial fungi) were the primary fungal isolates found, regardless of habitat differences. There was some variation in fungal components amongst colonies, even within the same apiary. The variable components were Absidia, Alternaria, Aureobasidium, Bipolaris, Fusarium, Geotrichum, Mucor, Nigrospora, Paecilomyces, Scopulariopsis, and Trichoderma. The number of fungal isolates was reduced as an effect of fungicide contamination. Aspergillus abundance was particularly affected by increased fungicide levels, as indicated by Simpson's diversity index. Bee bread showing fungicide contamination originated from colonies, many of which showed chalkbrood symptoms.

Water conservation features of ova of Agrilus planipennis (Coleoptera: Buprestidae).

The emerald ash borer, Agrilus planipennis Fairmaire, has destroyed millions of ash trees (Fraxinus spp.) in North America since first identified in Detroit in 2002. With species of ash distributed throughout North America, it is easy to speculate the extinction of all susceptible species of ash on the continent given a lack of physical, environmental, or climactic barrier for dispersal of the insect. We investigated water balance characteristics of emerald ash borer ova by using gravimetric methods in an effort to measure their response to heat- and water-stress and explore possible influences this stress may have on the ecology and physiology of the ovum. We also explored the possible water balance benefit of a peculiar, "clustering," oviposition behavior, as well as the difference in responses to stress between ova from a laboratory colony and ova from two wild populations. We found no evidence of water vapor absorption as a water balance strategy; rather enhanced water retention, resistance to desiccation, and viability with low water content were important survival strategies for these ova. Surface lipids resist thermal breakdown as indicated by ova having no detectable critical transition temperature, maintaining their water-proofing function as temperature rises. The observed "clustering" behavior had no desiccation-avoidance benefit and ova from the wild populations behaved almost identically to the ova from the lab colony, although the lab ova were slightly larger and more sensitive to dehydration. Given this new information, there appears to be no heat- or water-stress barriers for the dispersal of this devastating pest at the ovum stage.

Water balance characteristics of pupae developing in different size maggot masses from six species of forensically important flies.

The impact of maggot mass size on body water content, net transpiration rate, and dehydration tolerance of fly pupae was examined in six species of necrophagous flies. Species that spent more time on food as larvae, produced pupae with high body water contents. Dehydration tolerance limits of pupae were modest, matching the moisture-rich conditions of decaying carrion for larvae. Protophormia terraenovae pupariates on food as it dries, and this was reflected by pupae having the highest body water content and lowest net transpiration rate. Megaselia scalaris featured the lowest body water content and highest dehydration tolerance, implying that this species is arid-suited, which matches its ability to feed and colonize on post-decay carrion. Lucilia illustris was the most sensitive to larval overcrowding, resulting in a dramatic decrease in pupal size, early dispersal from food, fed less and had fast net transpiration rates. By contrast, Lucilia sericata was the most resistant, by showing no pupal size decrease and no change in net transpiration rate. Other species were between these extremes, requiring larger maggot mass sizes to produce the effect of decreasing pupal size and increasing net transpiration rate. We conclude: (1) the pupa's response to overcrowding and water balance profile are species-specific, varying according to pupal size and net transpiration rate as independent characteristics; (2) water balance profile of the pupae reflects the behavior and microhabitat of the larva; and (3) danger of lethal desiccation to smaller-sized pupae is circumvented by a faster developmental rate rather than enhanced water conservation.

Multiple traumatic insemination events reduce the ability of bed bug females to maintain water balance.

To examine how traumatic insemination, a wounding process to females inflicted by males during copulation, reduces the longevity of females of the bedbug, Cimex lectularius, we assessed if multiple bouts of mating impact water relations of females by measuring net transpiration water loss rates. Our studies show that net transpiration rate of females correlates with frequency of mating (small increase after exposure to low numbers of males; large increase after exposure to large numbers of males), and this is reflected by reduced female survivorship for as much as 22 days at 75% RH, 25°C. Water loss occurs up to 28% more rapidly in females after being held with large groups of males. Females that were exposed to males having their paramere removed, females exposed only to other females, and females kept in isolation (unmated) exhibited no reduction in ability to retain water, indicating that traumatic insemination was responsible for the net transpiration rate increase. Mechanical piercing of the female's abdominal wall leads to increased net transpiration rates for longer periods than puncturing the ectospermalege (regular mating site), implying that inaccurate copulation by males is extremely detrimental to the water balance of females and that the ectospermalege is uniquely modified to seal off more quickly to prevent excess water loss. Mating frequency and the associated increased water loss is considerably reduced by the addition of bed bug alarm pheromone components. Thus, females experience elevated water stress due to traumatic insemination, especially at high levels and when males fail to pierce the ectospermalege, and water loss prevention, likely by more rapid sealing of the wound, is a novel function of the ectopsermalege.

Increased cave dwelling reduces the ability of cave crickets to resist dehydration.

Differential strategies for maintaining water balance are reported for female adults of three cave crickets Hadenoecus cumberlandicus, H. opilionoides and H. jonesi, a species replacement series along the Cumberland Plateau in the southeastern United States. The distribution of H. cumberlandicus is much broader than the range of H. opilionoides, which is much smaller in body size, and that of H. jonesi, which possesses enhanced troglomorphic (cave dwelling) characteristics. Due to high net transpiration (water loss) rates and increased activation energies, H. jonesi and H. opilionoides are more susceptible to dehydration than H. cumberlandicus. To avoid dehydration, H. opilionoides and H. jonesi require more moisture than H. cumberlandicus to counter their higher rates of water loss. The heightened reliance on moisture likely indicates that the more troglomorphic H. jonesi and smaller H. opilionoides are required to spend more time in the moist cave region. Reliance on the cave for H. cumberlandicus is presumably less, allowing them to function in epigean habitats for longer periods and disperse to nearby caves, likely accounting for the more expansive distribution of this cricket. While in the cave habitat, cave crickets are exposed to water-saturated conditions, reducing the pressure of dehydration stress the longer a species remains in this wet environment. This reduced pressure leads to higher water loss rates as cave confinement increases. We conclude that increasing water loss rates associated with increasing troglomorphic adaptation in cave crickets is a side effect of extended residence in stable moist cave environments.

The pheromone of the cave cricket, Hadenoecus cumberlandicus, causes cricket aggregation but does not attract the co-distributed predatory spider, Meta ovalis.

Food input by the cave cricket, Hadenoecus cumberlandicus Hubble & Norton (Orthoptera: Rhaphidophoridae), is vital to the cave community, making this cricket a true keystone species. Bioassays conducted on cave walls and in the laboratory show that clustering in H. cumberlandicus is guided by a pheromone, presumably excreta. This aggregation pheromone was demonstrated by using filter paper discs that had previous adult H. cumberlandicus exposure, resulting in > 70% response by either nymphs or adults, prompting attraction (thus, active component is a volatile), followed by reduced mobility (arrestment) on treated surfaces. Adults were similarly responsive to pheromone from nymphs, agreeing with mixed stage composition of clusters in the cave. Effects of [0.001 M - 0.1 M] uric acid (insect excreta's principle component) on H. cumberlandicus behavior were inconsistent. This pheromone is not a host cue (kairomone) and is not used as a repellent (allomone) as noted through lack of responses to natural H. cumberlandicus pheromone and uric acid concentrations by a co-occurring predatory cave orb weaver spider, Meta ovalis Gertsch (Araneae: Tetragnathidae). This pheromone is not serving as a sex pheromone because nymphs were affected by it and because this population of H. cumberlandicus is parthenogenic. The conclusion of this study is that the biological value of the aggregation pheromone is to concentrate H. cumberlandicus in sheltered sites in the cave conducive for minimizing water stress. Rather than signaling H. cumberlandicus presence and quality, the reduced mobility expressed as a result of contacting this pheromone conceivably may act as a defense tactic (antipredator behavior) against M. ovalis, which shares this favored habitat site.

Use of an alarm pheromone against ants for gaining access to aphid/scale prey by the red velvet mite Balaustium sp. (Erythraeidae) in a honeydew-rich environment.

This study shows that honeydew prompts arrestment and reduced activity, but not attraction, by the mite Balaustium sp. nr. putmani. When presented with short-range, two-choice bioassays, mites ceased their characteristic rapid crawling activity when they encountered honeydew-treated surfaces, resulting in them clustering around the honeydew. Approximately 80% of mites were retained by honeydew, with responses being independent of both mite life-history stage and source of honeydew (coccid scale insect or aphid). No obvious crawling movements or redirection of running path were made to the honeydew by the mites, implying the lack of any kind of attractant. Response of mites to single-sugar presentations of the main honeydew components--glucose, sucrose, fructose and trehalose--(0.001-0.1 mmol l(-1)) were inconsistent and failed to reproduce the arrestment/clustering associated with raw honeydew, suggesting that none of these sugars is an active arrestant ingredient. Formation of feeding clusters on honeydew does not contribute to enhancing water conservation by suppressing net transpiration (water loss) rates of individual mites as group size increases, indicating that the clustering is an artifact of arrestment. We hypothesize that release of neryl formate by the mites reduces negative interactions with the local ant species commonly associated with honeydew. We hypothesize that honeydew serves as: (1) a cue that facilitates discovery of scale/aphid prey; (2) a retainer on plants where these prey are present, signaling abundance and quality; and (3) an alternative and supplemental food source like that noted for other plant-inhabiting predatory mites. Neryl formate serves as an alarm pheromone and foul-tasting allomonal defense secretion that prevents predation of mites by ants that co-exist with aphid/scale insects in these honeydew-rich habitats.

Addition of alarm pheromone components improves the effectiveness of desiccant dusts against Cimex lectularius.

We demonstrate that the addition of bed bug, Cimex lectularius, alarm pheromone to desiccant formulations greatly enhances their effectiveness during short-term exposure. Two desiccant formulations, diatomaceous earth (DE) and Dri-die (silica gel), were applied at the label rate with and without bed bug alarm pheromone components, (E)-2-hexenal, (E)-2-octenal, and a (E)-2-hexenal:(E)-2-octenal blend. First-instar nymphs and adult females were subjected to 10-min exposures, and water loss rates were used to evaluate the response. Optimal effectiveness was achieved with a pheromone concentration of 0.01 M. With Dri-die alone, the water loss was 21% higher than in untreated controls, and water loss increased nearly two times with (E)-2-hexenal and (E)-2-octenal and three times with the (E)-2-hexenal: (E)-2-octenal blend. This shortened survival of first-instar nymphs from 4 to 1 d, with a similar reduction noted in adult females. DE was effective only if supplemented with pheromone, resulting in a 50% increase in water loss over controls with the (E)-2-hexenal:(E)-2-octenal blend, and a survival decrease from 4 to 2 d in first-instar nymphs. Consistently, the addition of the pheromone blend to desiccant dust was more effective than adding either component by itself or by using Dri-die or DE alone. Based on observations in a small microhabitat, the addition of alarm pheromone components prompted bed bugs to leave their protective harborages and to move through the desiccant, improving the use of desiccants for control. We concluded that short exposure to Dri-die is a more effective treatment against bed bugs than DE and that the effectiveness of the desiccants can be further enhanced by incorporation of alarm pheromone. Presumably, the addition of alarm pheromone elevates excited crawling activity, thereby promoting cuticular changes that increase water loss.

High temperature effects on water loss and survival examining the hardiness of female adults of the spider beetles, Mezium affine and Gibbium aequinoctiale.

A remarkable ability to tolerate temperatures as high as 52 degrees C for Mezium affine Boieldieu and 56 degrees C for Gibbium aequinoctiale Boieldieu (Coleoptera: Anobiidae) was discovered as part of a water balance study that was conducted to determine whether desiccation-resistance (xerophilic water balance classification) is linked to survival at high temperature. Characteristics of the heat shock response were an intermediate, reversible level of injury, appearing as though dead; greater recovery from heat shock by G. aequinoctiale (57%) than M. affine (30%) that supplemented higher temperature survival by G. aequinoctiale; and lack of protection generated by conditioning at sublethal temperature. Heatinduced mortality is attributed to an abrupt, accelerated water loss at 50 degrees C for M. affine and 54 degrees C for G. aequinoctiale, not to the species (M. affine) that loses water the slowest and has the lower activation energy, E(a) as a measure of cuticular boundary effectiveness. These temperatures where water loss increases sharply are not critical transition temperatures because Arrhenius analysis causes them to be erased (uninterrupted Boltzmann function) and E(a) fails to change when cuticular lipid from these beetles is removed. Our conclusion is that the temperature thresholds for survival and accelerated water loss closely match, and the key survival element in hot and dry environments contributing to wide distribution of G. aequinoctiale and M. affine derives from rising temperature prompting entry into quiescence and a resistance in cuticular lipid fluidity.

The giant Madagascar hissing-cockroach (Gromphadorhina portentosa) as a source of antagonistic moulds: concerns arising from its use in a public setting.

Cockroaches and their excreta have been linked to allergies and childhood asthma. The giant Madagascar hissing-cockroach (Gromphadorhina portentosa), heralded as an educational tool in classrooms, museums, zoos, is now available to the public as a children's pet raising health concerns. We provide a catalogue of potentially antagonistic moulds associated with this insect. Specimens were obtained from laboratories, classrooms, pet stores and private homes. Three different agar media were used to culture moulds from both internal and external cockroach surfaces. Pure cultures were obtained from hyphal tips excised by scalpel. A total of 14 mould taxa were obtained, mostly from external surfaces. The mycoflora was dominated by species of Rhizopus, Penicillium, Mucor, Trichoderma and Alternaria, and differed little among nymphs, adults, cast skins (exuviae) and faeces. A two-fold increase of Aspergillus niger isolates, however, was detected in exuviae and faeces. The mycoflora appeared to be equally distributed on the body regions in nymphs and adults alike. Most of the moulds recovered are common, well-known saprophytes with a prolific ability to produce asexual spores (e.g. conidia) when supplied with adequate moisture and an organic substrate (e.g. vegetable matter, pet food and exuviae). Cockroach rearing conditions thus serve as an ideal environment for mould growth and proliferation, and the subsequent use (handling) of these insects in a public forum increases the risk of inducing mould-related allergies in humans. Of special concern are moulds also capable of causing secondary infections (e.g. Rhizopus, Mucor, Aspergillus), gaining entry via open wounds and inhalation. This is mainly a point of public awareness aimed at individuals (especially children) prone to infections and allergies that might be exposed to this insect and/or its rearing conditions.

An endosymbiotic conidial fungus, Scopulariopsis brevicaulis, protects the American dog tick, Dermacentor variabilis, from desiccation imposed by an entomopathogenic fungus.

The functional role of an endosymbiotic conidial fungus (Scopulariopsis brevicaulis) prevalent within the integumental glands and hemocoel of the American dog tick (Dermacentor variabilis) was investigated to explore the nature of this tick/fungus association. D. variabilis is normally highly resistant to Metarhizium anisopliae, a widely-distributed entomopathogenic fungus, but when mature female ticks harboring S. brevicaulis were fed a solution containing a mycotoxin (Amphotericin B) to purge this mycobiont internally, the ticks inoculated with M. anisopliae displayed classic signs of pathogenicity, as evidenced by recovery of M. anisopliae from ticks by internal fungus culture, greatly accelerated net transpiration water loss rates (nearly 3x faster than ticks containing S. brevicaulis naturally) and elevation of critical equilibrium humidity (CEH) closer to saturation, implying a reduced capacity to absorb water vapor and disruption of water balance (water gain not equal water loss) that resulted in tick death. The presence of S. brevicaulis within the tick was previously puzzling: the fungus is transmitted maternally and there is no apparent harm inflicted to either generation. This study suggests that S. brevicaulis provides protection to D. variabilis ticks against M. anisopliae. Thus, the S. brevicaulis/tick association appears to be mutualistic symbiosis. Given that both organisms are of medical-veterinary importance, disruption of this symbiosis has potential for generating novel tools for disease control.

Resistance to dehydration between bouts of blood feeding in the bed bug, Cimex lectularius, is enhanced by water conservation, aggregation, and quiescence.

To determine how the bed bug, Cimex lectularius, survives in a dry environment for many months without feeding, water-balance characteristics were compared for all stages from first-instar nymphs to adults. This species is characterized by a low net transpiration rate averaging < 0.2%/h, high tolerance for dehydration (30-40% loss in body water), and an impermeable cuticle as indicated by a high critical transition temperature (CTT) in the 35-40 degrees C range, implying that this insect is adapted for desiccation-hardiness. The capacity of adults to survive for 2 weeks at 0.00a(v) (a(v) = % RH/100) with no access to food or water exemplifies this trait. In contrast to more mature stages, first-instar nymphs contain more water, lose water at a faster rate, experience abrupt water loss at a lower temperature, and survive less time in dry air, suggesting that this stage is the most sensitive to water stress. This insect relies on blood to replenish water stores; none of the stages examined have the capacity to absorb water vapor (critical equilibrium activity, CEA > or = 0.99a(v)), and they drank only sparingly when offered free water. As the bed bugs progress through their development, they gradually reduce their water requirements while increasing their desiccation resistance. Surviving water stress is considerably enhanced behaviorally by quiescence, characterized by prolonged periods of inactivity, and by the formation of clusters that generate a water-conserving group effect.

Allergenicity of the Madagascar hissing cockroach.

BACKGROUND: The Madagascar hissing cockroach (Gromphadorhina portentosa) has become popular as a pet and as an educational tool in classrooms, zoos, and museums. OBJECTIVE: To determine whether proteins in G. portentosa are allergens and whether these allergens cross-react with those of other cockroach species. METHODS: Sera from cockroach-sensitive individuals and control subjects were used to probe immunoblots for the presence of circulating IgE that bound to proteins present in extracts of 4 cockroach species. RESULTS: Serum from an individual sensitized to G portentosa had circulating IgE that bound to proteins in extracts of all 4 cockroach species. Eight of 15 Blatella germanica-sensitized patients had IgE that bound to proteins in extracts prepared from G portentosa. Rabbit antiserum to Periplaneta americana and to Bla g 1 also contained antibody that bound to proteins in G. portentosa extract, demonstrating antigenic cross-reactivity among these cockroach species. CONCLUSIONS: Allergists, teachers, parents, and patients should be aware that sensitization and allergic reactions to the Madagascar hissing cockroach can occur. Caution should be taken if these exotic cockroaches are used in educational exercises in schools, museums, and zoos to protect predisposed individuals.

Evidence of a maternal effect that protects against water stress in larvae of the American dog tick, Dermacentor variabilis (Acari: Ixodidae).

We report that the ability to absorb water vapor from the air in larvae of the American dog tick, Dermacentor variabilis, changes depending upon moisture conditions where the eggs develop. When development occurs at lower relative humidities, resultant larvae can replenish water stores, maintain water balance, and survive at relative humidities as low as 75-85% RH, a range that agrees with previously published values for the critical equilibrium humidity or CEH. In contrast, exposure to high relative humidity conditions during development elevates the CEH to 93-97% RH. These larvae can survive only at relative humidities that are close to saturation, as 93% RH is a dehydrating atmosphere. For these larvae, absorption at 97% RH can be prevented by blocking the mouthparts with wax, indicating that an upward shift has occurred in the moisture threshold where the active mechanism for water vapor absorption operates. Based on transfer experiments between low and high relative humidities, the CEH of larvae is determined by the relative humidity experienced by the mother rather than the moisture conditions encountered by eggs after they are laid. The fact that no changes in body water content, dehydration tolerance limit and water loss rate were observed implies that adjustments to the CEH conferred by the mother have the adaptive significance of enabling larvae to maintain water balance by limiting the range of hydrating atmospheres.

Stress-induced accumulation of glycerol in the flesh fly, Sarcophaga bullata: evidence indicating anti-desiccant and cryoprotectant functions of this polyol and a role for the brain in coordinating the response.

Nondiapausing larvae of the flesh fly, Sarcophaga bullata, responded to several forms of short-term environmental stress (low temperature, anoxia and desiccation) by accumulating glycerol. Elevation of this polyol, regardless of the type of stress that induced accumulation, conferred cold resistance: larvae with high glycerol levels were 3-4 times more tolerant of a 2h exposure to -10 degrees C than unstressed larvae. Protection against low temperature injury, as well as dehydration, was also attained by injection of exogenous glycerol into third instar larvae. This artificially induced cold hardiness was only temporary: when glycerol-injected larvae were exposed to -10 degrees C immediately after injection, survival was high, but none survived if they were injected and then held at 25 degrees C for 2 days before the -10 degrees C exposure. Larvae ligated behind the brain immediately after low temperature exposure failed to accumulate glycerol, but glycerol did accumulate in larvae ligated 6-24h after cold treatment, thus implying a critical role for the brain in initiating glycerol production. Interestingly, a much shorter exposure (2h) to low temperature was sufficient to reduce the maximum rate of water loss. Collectively, these observations suggest that multiple pathways may be exploited in response to stress: one pathway is most likely associated with rapid cold hardening (RCH) which generates immediate protection, and a second pathway remains activated for a longer period to enhance the initial protection afforded by glycerol.