Carvacrol and Thymol Combat Desiccation Resistance Mechanisms in Salmonella enterica Serovar Tennessee

Some Salmonella enterica serovars are frequently associated with disease outbreaks in low-moisture foods (LMF) due to their ability to adapt efficiently to desiccation stress. These serovars are often persistent during food processing. Disruption of these resistance responses was accomplished previously using the membrane-active lipopeptide, paenibacterin. This study was initiated to determine how desiccation resistance mechanisms are overcome when Salmonella Tennessee, a known resistant serovar, is treated with the membrane-active food additives carvacrol and thymol. Knowing that the minimum inhibitory concentrations (MICs) of carvacrol and thymol against Salmonella Tennessee are 200 and 100 µg/mL, the concentrations tested were 100–400 and 50–200 µg/mL, respectively. Results show that desiccation-adapted Salmonella Tennessee, prepared by air drying at 40% relative humidity and 22–25 °C for 24 h, was not inactivated when exposed for 4.0 h to less than 2xMIC of the two additives. Additionally, treatment of desiccation-adapted Salmonella Tennessee for 120 min with carvacrol and thymol at the MIC-level sensitized the cells (1.4–1.5 log CFU/mL reduction) to further desiccation stress. Treating desiccation-adapted Salmonella Tennessee with carvacrol and thymol induced leakage of intracellular potassium ions, reduced the biosynthesis of the osmoprotectant trehalose, reduced respiratory activity, decreased ATP production, and caused leakage of intracellular proteins and nucleic acids. Carvacrol, at 200–400 µg/mL, significantly downregulated the transcription of desiccation-related genes (proV, STM1494, and kdpA) as determined by the reverse-transcription quantitative PCR. The current study revealed some of the mechanisms by which carvacrol and thymol combat desiccation-resistant Salmonella Tennessee, raising the feasibility of using these additives to control desiccation-adapted S. enterica in LMF.

Yellow-y Functions in Egg Melanization and Chorion Morphology of the Asian Tiger Mosquito, Aedes albopictus

The Asian tiger mosquito, Aedes albopictus, is one of the most serious public health pests, which can transmit various vector-borne diseases. Eggs from this mosquito species become dark black shortly after oviposition and exhibit high desiccation resistance. Some of the Yellow proteins that act as dopachrome conversion enzymes (DCEs) are involved in the tyrosine-mediated tanning (pigmentation and sclerotization) metabolic pathway that significantly accelerates melanization reactions in insects. In this research, we analyzed the function of one of the yellow genes, yellow-y (AalY-y), in eggshell/chorion melanization of Ae. albopictus eggs. Developmental and tissue-specific expression measured by real-time PCR showed that AalY-y transcripts were detected at all stages of development analyzed, with significantly higher levels in the ovaries from blood-fed adult females. Injection of double-stranded RNA for AalY-y (dsAalY-y) had no significant effect on fecundity. However, unlike dsEGFP-treated control eggs that become black by 2–3 h after oviposition (HAO), dsAalY-y eggs were yellow-brown at 2 HAO, and reddish-brown even at 48 HAO. dsEGFP eggs exhibited resistance to desiccation at 48 HAO, whereas approximately 50% of the dsAalY-y eggs collapsed when they were moved to a low humidity condition. In addition, TEM analysis revealed an abnormal morphology and ultrastructure of the outer-endochorion in the dsAalY-y eggs. These results support the hypothesis that AalY-y is involved in the tyrosine-induced melanin biosynthetic pathway, plays an important role in black melanization of the chorion and functions in conferring proper morphology of the outer-endochorion, a structure that is presumably required for egg desiccation resistance in Ae. albopictus.

Physiological, morphological and ecological traits drive desiccation resistance in north temperate dung beetles

Background Increasing temperatures and changes in precipitation patterns threaten the existence of many organisms. It is therefore informative to identify the functional traits that underlie differences in desiccation resistance to understand the response of different species to changes in water availability resulting from climate change. We used adult dung beetles as model species due to their importance to ecosystem services. We investigated: (i) the effect of physiological (water loss rate, water loss tolerance, body water content), morphological (body mass) and ecological (nesting behaviour) traits on desiccation resistance; (ii) the role of phylogenetic relatedness in the above associations; and, (iii) whether relatively large or small individuals within a species have similar desiccation resistance and whether these responses are consistent across species. Results Desiccation resistance decreased with increasing water loss rate and increased with increasing water loss tolerance (i.e. proportion of initial water content lost at the time of death). A lack of consistent correlation between these traits due to phylogenetic relatedness suggests that the relationship is not determined by a shared evolutionary history. The advantage of a large body size in favouring desiccation resistance depended on the nesting behaviour of the dung beetles. In rollers (one species), large body sizes increased desiccation resistance, while in tunnelers and dwellers, desiccation resistance seemed not to be dependent on body mass. The phylogenetic correlation between desiccation resistance and nesting strategies was significant. Within each species, large individuals showed greater resistance to desiccation, and these responses were consistent across species. Conclusions Resistance to desiccation was explained mainly by the dung beetles’ ability to reduce water loss rate (avoidance) and to tolerate water loss (tolerance). A reduction in water availability may impose a selection pressure on body size that varies based on nesting strategies, even though these responses may be phylogenetically constrained. Changes in water availability are more likely to affect dweller species, and hence the ecosystem services they provide.

Effects of Thermal Acclimation on the Tolerance of Bactrocera zonata (Diptera: Tephritidae) to Hydric Stress

Insects, similarly to other small terrestrial invertebrates, are particularly susceptible to climatic stress. Physiological adjustments to cope with the environment (i.e., acclimation) together with genetic makeup eventually determine the tolerance of a species to climatic extremes, and constrain its distribution. Temperature and desiccation resistance in insects are both conditioned by acclimation and may be interconnected, particularly for species inhabiting xeric environments. We determined the effect of temperature acclimation on desiccation resistance of the peach fruit fly (Bactrocera zonata, Tephritidae) – an invasive, polyphagous pest, currently spreading through both xeric and mesic environments in Africa and the Eurasian continent. Following acclimation at three constant temperatures (20, 25, and 30°C), the survival of adult flies deprived of food and water was monitored in extreme dry and humid conditions (<10 and >90% relative humidity, respectively). We found that flies acclimated at higher temperatures were significantly heavier, and contained more lipids and protein. Acclimation temperature significantly and similarly affected the survival of males and females at both high and low humidity conditions. In both cases, flies maintained at 30°C survived longer compared to 20 and 25°C – habituated counterparts. Regardless of the effect of acclimation temperature on survival, overall life expectancy was significantly shortened when flies were assayed under desiccating conditions. Additionally, our experiments indicate no significant difference in survival patterns between males and females, and that acclimation temperature had similar effects after both short (5–10 days) and long (11–20 days) acclimation periods. We conclude that acclimation at 30°C prolongs the survival of B. zonata, regardless of ambient humidity levels. Temperature probably affected survival through modulating feeding and metabolism, allowing for accumulation of larger energetic reserves, which in turn, promoted a greater ability to resist starvation, and possibly desiccation as well. Our study set the grounds for understanding the phenotypic plasticity of B. zonata from the hydric perspective, and for further evaluating the invasion potential of this pest.

Relationship of growth conditions to desiccation tolerance of Salmonella enterica, Escherichia coli, and Listeria monocytogenes

Growth on solid media as sessile cells is believed to increase the desiccation tolerance of Salmonella enterica . However, the reasons behind increased resistance have not been well explored. In addition, the same effect has not been examined for other foodborne pathogens such as pathogenic Escherichia coli or Listeria monocytogenes . The purpose of this research was two-fold: first, to determine the role of oxygenation during growth on the desiccation resistance of S. enterica , E. coli , and L. monocytogenes , and second, to determine the effect of sessile versus planktonic growth on the desiccation resistance of these pathogens. Three different serotypes each of Salmonella , E. coli , and L. monocytogenes were cultured in trypticase soy broth with 0.6% yeast extract (TSBYE), with (aerobic) shaking or on TSBYE with agar (TSAYE) under either aerobic or anaerobic conditions and harvested in stationary phase. After adding cell suspensions to cellulose filter disks, pathogen survival was determined by enumeration at 0 and after drying for 24 h. Results showed statistical differences in harvested initial populations prior to drying (0 h). For Salmonella , a correlation was found between high initial population and greater survival on desiccation (p = 0.05). In addition, statistical differences (p ≤ 0.05) between survival based on growth type were identified. However, differences found were not the same for the three pathogens, or between their serotypes. In general, Salmonella and E. coli desiccation resistance followed the pattern of aerobic agar media ≥ liquid media ≥ anaerobic agar media. For L. monocytogenes serotypes, resistance to desiccation was not statistically different based on mode of growth. These results indicate growth on solid media under aerobic conditions is not always necessary for optimal desiccation survival but may be beneficial when the desiccation resistance of the test serotype is unknown.

Desiccation stress acts as cause as well as cost of dispersal in Drosophila melanogaster

Environmental stress is one of the important causes of biological dispersal. At the same time, the process of dispersal itself can incur and/or increase susceptibility to stress for the dispersing individuals. Therefore, in principle, stress can serve as both a cause and a cost of dispersal. Desiccation stress is an environmentally relevant stress faced by many organisms, known to shape their population dynamics and distribution. However, the potentially contrasting roles of desiccation stress as a cause and a cost of dispersal have not been investigated. Furthermore, while desiccation stress often affects organisms in a sex-biased manner, it is not known whether the desiccation-dispersal relationship varies between males and females. We studied the role of desiccation stress as a cause and cost of dispersal in a series of experiments using D. melanogaster adults in two-patch dispersal setups. We were interested in knowing whether (a) dispersers are the individuals that are more susceptible to desiccation stress, (b) dispersers pay a cost in terms of reduced resistance to desiccation stress, (c) dispersal evolution alters the desiccation cost of dispersal, and (d) females pay a reproductive cost of dispersal. For this, we modulated the degree of desiccation stress faced by the flies as well as the provision of rest following a dispersal event. Our data showed that desiccation stress served as a significant cause of dispersal in both sexes. Further investigation revealed an increase in both male and female dispersal propensity with increasing desiccation duration. Next, we found a male-biased cost of dispersal in terms of reduced desiccation resistance. This trend was preserved in dispersal-selected and non-selected controls as well, where the desiccation cost of dispersal in females was very low compared to the males. Finally, we found that the females instead paid a significant reproductive cost of dispersal. Our results highlight the complex relationship between desiccation stress and dispersal, whereby desiccation resistance can show both a positive and a negative association with dispersal. Furthermore, the sex differences observed in these trait associations may translate into differences in movement patterns, thereby giving rise to sex-biased dispersal.

Cuticle Hydrocarbons Show Plastic Variation under Desiccation in Saline Aquatic Beetles

In the context of aridification in Mediterranean regions, desiccation resistance and physiological plasticity will be key traits for the persistence of aquatic insects exposed to increasing desiccation stress. Control of cuticular transpiration through changes in the quantity and composition of epicuticular hydrocarbons (CHCs) is one of the main mechanisms of desiccation resistance in insects, but it remains largely unexplored in aquatic ones. We studied acclimation responses to desiccation in adults of two endemic water beetles from distant lineages living in Mediterranean intermittent saline streams: Enochrus jesusarribasi (Hydrophilidae) and Nebrioporus baeticus (Dytiscidae). Cuticular water loss and CHC composition were measured in specimens exposed to a prior non-lethal desiccation stress, allowed to recover and exposed to a subsequent desiccation treatment. E. jesusarribasi showed a beneficial acclimation response to desiccation: pre-desiccated individuals reduced cuticular water loss rate in a subsequent exposure by increasing the relative abundance of cuticular methyl-branched compounds, longer chain alkanes and branched alkanes. In contrast, N. baeticus lacked acclimation capacity for controlling water loss and therefore may have a lower physiological capacity to cope with increasing aridity. These results are relevant to understanding biochemical adaptations to drought stress in inland waters in an evolutionary and ecological context.