Desiccation Survival Research Articles

Gut yeasts do not improve desiccation survival in Drosophila melanogaster

A healthy gut microbiota generally improves the performance of its insect host. Although the effects can be specific to the species composition of the microbial community, the role of gut microbiota in determining water balance has not been well explored. We used axenic and gnotobiotic (reared with a known microbiota) Drosophila melanogaster to test three hypotheses about the effects of gut yeasts on the water balance of adult flies: 1) that gut yeasts would improve desiccation survival in adult flies; 2) that larval yeasts would improve adult desiccation survival; 3) that the effects would be species-specific, such that yeasts closely associated with D. melanogaster in nature are more likely to be beneficial than those rarely found in association with D. melanogaster. We used Saccharomyces cerevisiae (often used in Drosophila cultures, but rarely associated with D. melanogaster in nature), Lachancea kluyveri (associated with some species of Drosophila, but not D. melanogaster), and Pichia kluyveri (associated with D. melanogaster in nature). Adult inoculation with yeasts had no effect on survival of desiccating conditions. Inoculation with P. kluyveri as larvae did not change desiccation survival in adults; however, rearing with L. kluyveri or S. cerevisiae reduced adult desiccation survival. We conclude that adult inoculation with gut yeasts has no impact on desiccation survival, but that rearing with yeasts can have either no or detrimental effect. The effects appear to be species-specific: P. kluyveri did not have a negative impact on desiccation tolerance, suggesting some level of co-adaptation with D. melanogaster. We note that S. cerevisiae may not be an appropriate species for studying the effects of gut yeasts on D. melanogaster.

Changes in lipid classes of Drosophila melanogaster in response to selection for three stress traits

Laboratory selection on environmental stress traits is an evolutionary approach that is informative in the context of understanding stress adaptation. Here we characterize changes in a lipidome of Drosophila melanogaster in lines selected for increased heat (elevated heat knockdown refractoriness), cold (decreased time to recover from chill-coma) and desiccation survival. Selection for desiccation resistance resulted in changes in multiple lipid classes used to characterize a lipidome. This included a decrease in triacylglycerols (TAGs) which is relevant to interpretation of storage lipid levels in previous D. melanogaster desiccation survival selection experiments. Chill-coma recovery rate selection was expected to show extensive changes in lipid classes, but only phosphatidic acids exhibited significant change. Selection for increased heat knockdown resistance resulted in a substantial change in the abundance of a class of lipids (diacylglycerols) which could play a role in mediating the heat shock response or result in an increase in neutral lipid mobilization.

Desiccation, thermal stress and associated mortality in Drosophila fruit flies induced by neuropeptide analogue treatment

Drosophila suzukii is a serious pest of soft fruit worldwide. With the global over-dependence on broad-spectrum pesticides, a strong imperative exists for more environmentally friendly and targeted methods of control. One promising avenue involves employing synthetic neuropeptide analogues as insecticidal agents to reduce pest fitness. Neuropeptides, central to the regulation of physiological and behavioural processes, play a vital role in cold and desiccation survival. Building upon this, the current study investigated the effects of biostable kinin, the cardioacceleratory peptide CAP2b and pyrokinin (PK) analogues (the latter of which have previously displayed cross-talk with the capa receptor), on desiccation, starvation and cold stress tolerance of the pest, D. suzukii, and the closely related non-pest, D. melanogaster. Results demonstrated analogues of the superfamily (CAP2b and PK derived) significantly impacted survival of the target insect under conditions of desiccation stress. However, these peptides enhanced desiccation stress survival in relation to controls, suggesting that they may act as antagonists of the capa signalling pathway in the Malpighian tubules. Of particular note was the ability of analogues 1895 (2Abf-Suc-FGPRLa) and 1902 (2Abf-Suc-FKPRLa) to impact D. suzukii but not D. melanogaster. A focus on native Drosophila CAP2b/PK and kinin sequences in analogue development may yield pure agonists with diuretic action that may reduce desiccation stress survival in the pest flies. In highlighting the PRXamide neuropeptide superfamily more generally, and the structures of promising analogues more specifically, this research will feed the evolution of next-generation analogues and drive forward the development of neuropeptidomimetic-based agents.

Coping with low water activities and osmotic stress in Acinetobacter baumannii: significance, current status and perspectives.

Multidrug resistant (MDR) pathogens are one of the most pressing challenges of contemporary health care. Acinetobacter baumannii takes a predominant position, emphasized in 2017 by the World Health Organization. The increasing emergence of MDR strains strengthens the demand for new antimicrobials. Possible targets for such compounds might be proteins involved in resistance against low water activity environments, since A. baumannii is known for its pronounced resistance against desiccation stress. Despite the importance of desiccation resistance for persistence of this pathogen in hospitals, comparable studies and precise data on this topic are rare and the mechanisms involved are largely unknown. This review aims to give an overview of the studies performed so far and the current knowledge on genes and proteins important for desiccation survival. 'Osmotic stress' is not identical to 'desiccation stress', but the two share the response of bacteria to low water activities. Osmotic stress resistance is in general studied much better, and in recent years it turned out that accumulation of compatible solutes in A. baumannii comprises some special features such as the bifunctional enzyme MtlD synthesizing the unusual solute mannitol. Furthermore, the regulatory pathways, as understood today, will be discussed.

Desiccation Tolerance Assays for Acinetobacter baumannii.

Acinetobacter baumannii is a hospital-associated pathogen of growing importance and is a paradigm for endemic hospital contamination. Desiccation tolerance has been implicated as an important characteristic that potentiates the spread of A. baumannii in clinical settings through contaminated healthcare equipment and personnel. Desiccation is a multifaceted stress, and many physiological and environmental factors can influence its impact on bacterial survival. This chapter provides a protocol for assessing desiccation survival that facilitates comparisons among A. baumannii strains under various environmental conditions.

Not so free range? Oviposition microhabitat and egg clustering affects Eretmoptera murphyi (Diptera: Chironomidae) reproductive success

Understanding the physiology of non-native species in Antarctica is key to elucidating their ability to colonise an area, and how they may respond to changes in climate. Eretmoptera murphyi is a chironomid midge introduced to Signy Island (Maritime Antarctic) from South Georgia (Sub-Antarctic) where it is endemic. Here, we explore the tolerance of this species’ egg masses to heat and desiccation stress encountered within two different oviposition microhabitats (ground surface vegetation and underlying soil layer). Our data show that, whilst oviposition takes place in both substrates, egg sacs laid individually in soil are at the greatest risk of failing to hatch, whilst those aggregated in the surface vegetation have the lowest risk. The two microhabitats are characterised by significantly different environmental conditions, with greater temperature fluctuations in the surface vegetation, but lower humidity (%RH) and available water content in the soil. Egg sacs were not desiccation resistant and lost water rapidly, with prolonged exposure to 75% RH affecting survival for eggs in singly oviposited egg sacs. In contrast, aggregated egg sacs (n = 10) experienced much lower desiccation rates and survival of eggs remained above 50% in all treatments. Eggs had high heat tolerance in the context of the current microhabitat conditions on Signy. We suggest that the atypical (for this family) use of egg sac aggregation in E. murphyi has developed as a response to environmental stress. Current temperature patterns and extremes on Signy Island are unlikely to affect egg survival, but changes in the frequency and duration of extreme events could be a greater challenge.

The role of compatible solutes in desiccation resistance of Acinetobacter baumannii.

Acinetobacter baumannii is a nosocomial pathogen which can persist in the hospital environment not only due to the acquirement of multiple antibiotic resistances, but also because of its exceptional resistance against disinfectants and desiccation. A suitable desiccation assay was established in which A. baumannii ATCC 19606T survived for ca. 1 month. The growth medium slightly influenced survival after subsequent desiccation. A significant effect could be attributed to the growth phase in which bacteria were dried: In exponential phase, cells were much more desiccation sensitive. The main focus of the present study was the elucidation of the role of compatible solutes, which are known to protect many bacteria under low water activity conditions, in desiccation survival of A. baumannii. Exogenous trehalose was shown to efficiently protect A. baumannii on dry surfaces, in contrast to other compatible solutes tested such as mannitol or glycine betaine. To analyze the importance of intracellularly accumulated solutes, a double mutant lacking biosynthesis pathways for mannitol and trehalose was generated. This mutant accumulated glutamate as sole solute in the presence of high NaCl concentrations and showed severe growth defects under osmotic stress conditions. However, no effect on desiccation tolerance could be seen, neither when cells were dried in water nor in the presence of NaCl.

Cooption of heat shock regulatory system for anhydrobiosis in the sleeping chironomid Polypedilum vanderplanki

Polypedilum vanderplanki is a striking and unique example of an insect that can survive almost complete desiccation. Its genome and a set of dehydration-rehydration transcriptomes, together with the genome of Polypedilum nubifer (a congeneric desiccation-sensitive midge), were recently released. Here, using published and newly generated datasets reflecting detailed transcriptome changes during anhydrobiosis, as well as a developmental series, we show that the TCTAGAA DNA motif, which closely resembles the binding motif of the Drosophila melanogaster heat shock transcription activator (Hsf), is significantly enriched in the promoter regions of desiccation-induced genes in P. vanderplanki, such as genes encoding late embryogenesis abundant (LEA) proteins, thioredoxins, or trehalose metabolism-related genes, but not in P. nubifer Unlike P. nubifer, P. vanderplanki has double TCTAGAA sites upstream of the Hsf gene itself, which is probably responsible for the stronger activation of Hsf in P. vanderplanki during desiccation compared with P. nubifer To confirm the role of Hsf in desiccation-induced gene activation, we used the Pv11 cell line, derived from P. vanderplanki embryo. After preincubation with trehalose, Pv11 cells can enter anhydrobiosis and survive desiccation. We showed that Hsf knockdown suppresses trehalose-induced activation of multiple predicted Hsf targets (including P. vanderplanki-specific LEA protein genes) and reduces the desiccation survival rate of Pv11 cells fivefold. Thus, cooption of the heat shock regulatory system has been an important evolutionary mechanism for adaptation to desiccation in P. vanderplanki.

The Antioxidant System in the Anhydrobiotic Midge as an Essential, Adaptive Mechanism for Desiccation Survival.

One of the major damaging factors for living organisms experiencing water insufficiency is oxidative stress. Loss of water causes a dramatic increase in the production of reactive oxygen species (ROS). Thus, the ability for some organisms to survive almost complete desiccation (called anhydrobiosis) is tightly related to the ability to overcome extraordinary oxidative stress. The most complex anhydrobiotic organism known is the larva of the chironomid Polypedilum vanderplanki. Its antioxidant system shows remarkable features, such as an expansion of antioxidant genes, their overexpression, as well as the absence or low expression of enzymes required for the synthesis of ascorbate and glutathione and their antioxidant function. In this chapter, we summarize existing data about the antioxidant system of this insect, which is able to cope with substantial oxidative damage, even in an intracellular environment that is severely disturbed due to water loss.

Global Screening of Salmonella enterica Serovar Typhimurium Genes for Desiccation Survival.

Salmonella spp., one of the most common foodborne bacterial pathogens, has the ability to survive under desiccation conditions in foods and food processing facilities for years. This raises the concerns of Salmonella infection in humans associated with low water activity foods. Salmonella responds to desiccation stress via complex pathways involving immediate physiological actions as well as coordinated genetic responses. However, the exact mechanisms of Salmonella to resist desiccation stress remain to be fully elucidated. In this study, we screened a genome-saturating transposon (Tn5) library of Salmonella Typhimurium (S. Typhimurium) 14028s under the in vitro desiccation stress using transposon sequencing (Tn-seq). We identified 61 genes and 6 intergenic regions required to overcome desiccation stress. Salmonella desiccation resistance genes were mostly related to energy production and conversion; cell wall/membrane/envelope biogenesis; inorganic ion transport and metabolism; regulation of biological process; DNA metabolic process; ABC transporters; and two component system. More than 20% of the Salmonella desiccation resistance genes encode either putative or hypothetical proteins. Phenotypic evaluation of 12 single gene knockout mutants showed 3 mutants (atpH, atpG, and corA) had significantly (p < 0.02) reduced survival as compared to the wild type during desiccation survival. Thus, our study provided new insights into the molecular mechanisms utilized by Salmonella for survival against desiccation stress. The findings might be further exploited to develop effective control strategies against Salmonella contamination in low water activity foods and food processing facilities.

Renal neuroendocrine control of desiccation and cold tolerance by Drosophila suzukii.

BACKGROUNDNeuropeptides are central to the regulation of physiological and behavioural processes in insects, directly impacting cold and desiccation survival. However, little is known about the control mechanisms governing these responses in Drosophila suzukii. The close phylogenetic relationship of D. suzukii with Drosophila melanogaster allows, through genomic and functional studies, an insight into the mechanisms directing stress tolerance in D. suzukii.RESULTSCapability (Capa), leucokinin (LK), diuretic hormone 44 (DH44) and DH31 neuropeptides demonstrated a high level of conservation between D. suzukii and D. melanogaster with respect to peptide sequences, neuronal expression, receptor localisation, and diuretic function in the Malpighian tubules. Despite D. suzukii's ability to populate cold environments, it proved sensitive to both cold and desiccation. Furthermore, in D. suzukii, Capa acts as a desiccation‐ and cold stress‐responsive gene, while DH 44 gene expression is increased only after desiccation exposure, and the LK gene after nonlethal cold stress recovery.CONCLUSIONThis study provides a comparative investigation into stress tolerance mediation by neuroendocrine signalling in two Drosophila species, providing evidence that similar signalling pathways control fluid secretion in the Malpighian tubules. Identifying processes governing specific environmental stresses affecting D. suzukii could lead to the development of targeted integrated management strategies to control insect pest populations. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Desiccation tolerance of different life stages of the invasive marine kelp Undaria pinnatifida: Potential for overland transport as invasion vector

Overland transport in open-air-exposed conditions, including the translocation of specimens fouled on trailered boat hulls, aquaculture ropes or entangled in fishing gear, is recognized as a short-distance vector for the introduction of invasive aquatic species. Here, the desiccation tolerance for different life stages of the invasive kelp Undaria pinnatifida was determined to test the likelihood of air-exposed transport. Water content, photosynthetic quantum yield (Fv/Fm) as well as the capacity to release viable spores were monitored for mature sporophylls during a five-day exposure to air conditions. For newly settled spores (16h post-release) and developing gametophytes (30h post-release) survival and growth were observed after 1, 3, 6, 12 and 24h of desiccation. Additionally, spore settlement and desiccation survival (after 3, 12 and 48h) were determined for seven-day-old gametophytes growing on different rope materials (hemp, polypropylene, polyethylene, nylon) in high (99% relative air humidity, RH) and typical (59% RH) air humidity conditions. Viable spores were released from mature sporophylls after three days of desiccation. <0.4% of spores and gametophytes survived air exposure. Of those surviving, however, individual gametophytes could endure 12h of desiccation. These specimens exhibited enhanced average lengths, growing up to 70% larger than control gametophytes. Attached to ropes, gametophytes survived 48h of desiccation; settlement and desiccation survival did not differ between rope materials. Overall, this study demonstrates that air-exposed transport, especially the translocation of mature sporophylls, represents a potential spread mechanism for the invasive U. pinnatifida. This is a vector that should be considered for pest management.

Desiccation and ethanol resistances of multidrug resistant Acinetobacter baumannii embedded in biofilm: The favorable antiseptic efficacy of combination chlorhexidine gluconate and ethanol

Globally, multidrug-resistant Acinetobacter baumannii (MDRAB) has emerged as an important pathogen in nosocomial outbreaks. This study aimed to investigate the correlation between the biofilm formation and survival of MDRABs, and to investigate the antiseptic efficacy of hand sanitizers for the MDRABs, embedded with biofilm (MDRAB-Bs). The MDRABs were selected randomly after pulsed-field gel electrophoresis (PFGE), and their biofilm formation was analyzed. Desiccation and ethanol tolerances were assayed to test the bacterial survival. The antiseptic efficacy of combined chlorhexidine gluconate (CHG) and 70% ethanol agents against MDRAB-Bs were compared with the 70% ethanol cleanser. Eleven MDRABs, which varied in biofilm formation (MRDAB-B) and planktonic type (MDRAB-P), were tested. In desiccation survival, the mean survival time for the MDRAB-Bs was 49.0 days which was significantly higher than that of their planktonic type (17.3 days) (P<0.005). The MDRAB-Ps could be eliminated after a 10min contact with a 30% ethanol agent, however, it took 10min of 70% ethanol to eliminate the MDRAB-Bs. On the other hand, a 2% CHG in 70% ethanol solution completely eliminated all MDRAB-Bs after 1min contacted time. The 2% CHG in 70% ethanol agent provided a significantly superior efficacy than the 70% ethanol solution at eliminating the MDRAB-Bs (P<0.005). MDRAB with biofilm-formation presented significantly higher desiccation and ethanol resistances than their planktonic type. Moreover, the 2% CHG in 70% ethanol agent provided a superior antiseptic efficacy for MDRAB-Bs than that of the 70% ethanol agent.

Thermal Resistance and Gene Expression of both Desiccation-Adapted and Rehydrated Salmonella enterica Serovar Typhimurium Cells in Aged Broiler Litter.

The objective of this study was to investigate the thermal resistance and gene expression of both desiccation-adapted and rehydrated Salmonella enterica serovar Typhimurium cells in aged broiler litter. S Typhimurium was desiccation adapted in aged broiler litter with a 20% moisture content (water activity [aw], 0.81) for 1, 2, 3, 12, or 24 h at room temperature and then rehydrated for 3 h. As analyzed by quantitative real-time reverse transcriptase PCR (qRT-PCR), the rpoS, proV, dnaK, and grpE genes were upregulated (P < 0.05) under desiccation stress and could be induced after 1 h but in less than 2 h. Following rehydration, fold changes in the levels of these four genes became significantly lower (P < 0.05). The desiccation-adapted ΔrpoS mutant was less heat resistant at 75°C than was the desiccation-adapted wild type (P < 0.05), whereas there were no differences in heat resistance between desiccation-adapted mutants in two nonregulated genes (otsA and PagfD) and the desiccation-adapted wild type (P > 0.05). Survival characteristics of the desiccation-adapted ΔPagfD (rdar [red, dry, and rough] morphotype) and ΔagfD (saw [smooth and white] morphotype) mutants were similar (P > 0.05). Trehalose synthesis in the desiccation-adapted wild type was not induced compared to a nonadapted control (P > 0.05). Our results demonstrated the importance of the rpoS, proV, dnaK, and grpE genes in the desiccation survival of S Typhimurium. By using an ΔrpoS mutant, we found that the rpoS gene was involved in the cross-protection of desiccation-adapted S Typhimurium against high temperatures, while trehalose synthesis or rdar morphology did not play a significant role in this phenomenon. In summary, S Typhimurium could respond rapidly to low-aw conditions in aged broiler litter while developing cross-protection against high temperatures, but this process could be reversed upon rehydration.IMPORTANCE Physical heat treatment is effective in eliminating human pathogens from poultry litter used as biological soil amendments. However, prior to physical heat treatment, some populations of microorganisms may be adapted to the stressful conditions in poultry litter during composting or stockpiling, which may cross-protect them against subsequent high temperatures. Our previous study demonstrated that desiccation-adapted S. enterica cells in aged broiler litter exhibited enhanced thermal resistance. However, there is limited research on the underlying mechanisms of the extended survival of pathogens under desiccation conditions in animal wastes and cross-tolerance to subsequent heat treatment. Moreover, no information is available about the thermal resistance of desiccation-adapted microorganisms in response to rehydration. Therefore, in the present study, we investigated the gene expression and thermal resistance of both desiccation-adapted and rehydrated S Typhimurium in aged broiler litter. This work will guide future research efforts to control human pathogens in animal wastes used as biological soil amendments.

Phospholipase Dα1-mediated phosphatidic acid change is a key determinant of desiccation-induced viability loss in seeds.

High sensitivity of seeds to water loss is a widespread phenomenon in the world's plant species. The molecular basis of this trait is poorly understood but thought to be associated with critical changes in membrane function. We profiled membrane lipids of seeds in eight species with varying levels of desiccation tolerance and found a close association between reducing seed viability and increasing phosphatidic acid (PA). We applied hydration-dehydration cycles to Arabidopsis seeds, which are normally desiccation tolerant, to mimic the onset of desiccation sensitivity with progression towards germination and examined the role of phospholipase D (PLD) in desiccation stress-induced production of PA. We found that PLDα1 became more abundant and migrated from the cytosol to the membrane during desiccation, whereas PLDδ did not change, and that all desiccation-induced PA was derived from PLDα1 hydrolysis. When PLDα1 was suppressed, the germination level after each hydration-dehydration cycle improved significantly. We further demonstrated that PLDα1-mediated PA formation modulates desiccation sensitivity as applying its inhibitor improved seed desiccation tolerance and its suppression in protoplasts enhanced survival under dehydration. The insights provided by comparative lipidomics enable us to propose a new membrane-based model for seed desiccation stress and survival.

An Experimental Evolution Test of the Relationship between Melanism and Desiccation Survival in Insects.

We used experimental evolution to test the ‘melanism-desiccation’ hypothesis, which proposes that dark cuticle in several Drosophila species is an adaptation for increased desiccation tolerance. We selected for dark and light body pigmentation in replicated populations of D. melanogaster and assayed several traits related to water balance. We also scored pigmentation and desiccation tolerance in populations selected for desiccation survival. Populations in both selection regimes showed large differences in the traits directly under selection. However, after over 40 generations of pigmentation selection, dark-selected populations were not more desiccation-tolerant than light-selected and control populations, nor did we find significant changes in mass or carbohydrate amounts that could affect desiccation resistance. Body pigmentation of desiccation-selected populations did not differ from control populations after over 140 generations of selection, although selected populations lost water less rapidly. Our results do not support an important role for melanization in Drosophila water balance.

Molecular characterization of Tps1 and Treh genes in Drosophila and their role in body water homeostasis.

In insects, trehalose serves as the main sugar component of haemolymph. Trehalose is also recognized as a mediator of desiccation survival due to its proposed ability to stabilize membranes and proteins. Although the physiological role of trehalose in insects has been documented for decades, genetic evidence to support the importance of trehalose metabolism remains incomplete. We here show on the basis of genetic and biochemical evidence that the trehalose synthesis enzyme Tps1 is solely responsible for the de novo synthesis of trehalose in Drosophila. Conversely, a lack of the gene for the trehalose hydrolyzing enzyme Treh causes an accumulation of trehalose that is lethal during the pupal period, as is observed with Tps1 mutants. Lack of either Tps1 or Treh results in a significant reduction in circulating glucose, suggesting that the maintenance of glucose levels requires a continuous turnover of trehalose. Furthermore, changes in trehalose levels are positively correlated with the haemolymph water volume. In addition, both Tps1 and Treh mutant larvae exhibit a high lethality after desiccation stress. These results demonstrate that the regulation of trehalose metabolism is essential for normal development, body water homeostasis, and desiccation tolerance in Drosophila.

Desiccation survival time for eggs of a widespread and invasive Australian mosquito species, Aedes (Finlaya) notoscriptus (Skuse).

The Australian native mosquito Aedes (Finlaya) notoscriptus (Skuse) is closely associated with natural and artificial water holding receptacles. Eggs are laid in habitats where they are exposed to drying conditions as water levels fluctuate. Withstanding desiccation enables survival in challenging environments and increases the potential for establishment in non-native habitats. Until now, the desiccation resistance of Ae. notoscriptus eggs has been unknown despite the historical invasive success of this important dog heartworm and arbovirus vector. Viability and mean survival times of eggs from two Ae. notoscriptus populations (metropolitan areas of Sydney, NSW and Adelaide, SA) were evaluated, with eggs stored under three dryness conditions for up to 367 days. Our results revealed that Ae. notoscriptus eggs can withstand desiccation for extended periods, under a variety of conditions, with approximately 9-13% egg viability recorded after one year. This prolonged egg survival reflects the widespread distribution of this mosquito in Australia and its history of incursions and subsequent establishment in non-native habitats. Differences in mean egg volume were recorded in addition to significantly different egg length to width ratios for the two populations, which may reflect adaptation to biotope of origin and an associated likelihood of drought and drying conditions. The results of this study suggest that the desiccation resistant eggs of Ae. notoscriptus make this species highly adaptable, increasing the risk of movement to non-endemic regions of the world.

Reproductive arrest and stress resistance in winter-acclimated Drosophila suzukii

Overwintering insects must survive the multiple-stress environment of winter, which includes low temperatures, reduced food and water availability, and cold-active pathogens. Many insects overwinter in diapause, a developmental arrest associated with high stress tolerance. Drosophila suzukii (Diptera: Drosophilidae), spotted wing drosophila, is an invasive agricultural pest worldwide. Its ability to overwinter and therefore establish in temperate regions could have severe implications for fruit crop industries. We demonstrate here that laboratory populations of Canadian D. suzukii larvae reared under short-day, low temperature, conditions develop into dark ‘winter morph’ adults similar to those reported globally from field captures, and observed by us in southern Ontario, Canada. These winter-acclimated adults have delayed reproductive maturity, enhanced cold tolerance, and can remain active at low temperatures, although they do not have the increased desiccation tolerance or survival of fungal pathogen challenges that might be expected from a more heavily melanised cuticle. Winter-acclimated female D. suzukii have underdeveloped ovaries and altered transcript levels of several genes associated with reproduction and stress. While superficially indicative of reproductive diapause, the delayed reproductive maturity of winter-acclimated D. suzukii appears to be temperature-dependent, not regulated by photoperiod, and is thus unlikely to be ‘true’ diapause. The traits of this ‘winter morph’, however, likely facilitate overwintering in southern Canada, and have probably contributed to the global success of this fly as an invasive species.

Desiccation survival of Acinetobacter spp. in infant formula

Acinetobacter spp. are included under category B in the FAO-WHO list of organisms of concern for neonatal health following the consumption of powdered infant formula. However, the ability of Acinetobacter spp. to maintain their viability in desiccated infant formula over a storage period consistent with the shelf-life of commercially available powdered infant formula (2 years) has not been demonstrated. In this study, 9 clinical and food isolates of Acinetobacter baumannii, A. calcoaceticus, and Acinetobacter genomosp. 3 were desiccated in infant formula and then reconstituted at designated time points. Bacterial viability was followed for a maximum period of 24 months or until the strain became undetectable (<5 × 102 cfu/ml). For comparative purposes, one Enterobacter hormaechei and two Enterobacter cloacae strains were also monitored for their desiccation survival. The seven clinical and food strains remained cultivable for the whole duration of the study and showed biphasic survival curves. The initial drop in viable count was up to 3.5 log10 cfu/ml within 18 h of desiccation exposure. By the end of the study, the reduction in viability was between 3.6 and 4.8 log10 cfu/ml. In contrast the A. baumanii and A. calcoaceticus species type strains only persisted for 6 and 9 months, respectively, possibly due to laboratory adaptation. The E. cloacae and E. hormaechei strains were undetectable after 12 and 17 months, respectively. The persistence of Acinetobacter spp. strains in desiccated powdered infant formula, supports the FAO-WHO designation of this organism as a risk to neonatal health.