Anhydrobiotic State Research Articles

Desiccation survival in an Antarctic nematode: molecular analysis using expressed sequenced tags

BackgroundNematodes are the dominant soil animals in Antarctic Dry Valleys and are capable of surviving desiccation and freezing in an anhydrobiotic state. Genes induced by desiccation stress have been successfully enumerated in nematodes; however we have little knowledge of gene regulation by Antarctic nematodes which can survive multiple environmental stresses. To address this problem we investigated the genetic responses of a nematode species, Plectus murrayi, that is capable of tolerating Antarctic environmental extremes, in particular desiccation and freezing. In this study, we provide the first insight into the desiccation induced transcriptome of an Antarctic nematode through cDNA library construction and suppressive subtractive hybridization.ResultsWe obtained 2,486 expressed sequence tags (ESTs) from 2,586 clones derived from the cDNA library of desiccated P. murrayi. The 2,486 ESTs formed 1,387 putative unique transcripts of which 523 (38%) had matches in the model-nematode Caenorhabditis elegans, 107 (7%) in nematodes other than C. elegans, 153 (11%) in non-nematode organisms and 605 (44%) had no significant match to any sequences in the current databases. The 1,387 unique transcripts were functionally classified by using Gene Ontology (GO) hierarchy and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The results indicate that the transcriptome contains a group of transcripts from diverse functional areas. The subtractive library of desiccated nematodes showed 80 transcripts differentially expressed during desiccation stress, of which 28% were metabolism related, 19% were involved in environmental information processing, 28% involved in genetic information processing and 21% were novel transcripts. Expression profiling of 14 selected genes by quantitative Real-time PCR showed 9 genes significantly up-regulated, 3 down-regulated and 2 continuously expressed in response to desiccation.ConclusionThe establishment of a desiccation EST collection for Plectus murrayi, a useful model in assessing the structural, physiological, biochemical and genetic aspects of multiple stress tolerance, is an important step in understanding the genome level response of this nematode to desiccation stress. The type of transcript analysis performed in this study sets the foundation for more detailed functional and genome level analyses of the genes involved in desiccation tolerance in nematodes.

Mechanisms and molecular adaptation to extreme dehydration in tardigrades: Hsp gene expression in Milnesium tardigradum

The tardigrade Milnesium tardigradum Doyere 1840 is a well-known cosmopolitan species and a typical inhabitant of moist environments which facilitate the animal's gaseous exchange and avoid desiccation. However, such habitats frequently undergo seasonal changes that impact animal life. M. tardigradum is able to survive these periods of adverse conditions due to its ability to enter into a anhydrobiotic state. In adverse environments, all terrestrial and freshwater tardigades arrest their metabolic activity and get dehydrated to form the tun state. In the anhydrobiotic stage, tardigrades showextraordinary tolerance tophysical extremes includinghigh-energy radiation, immersion inorganic solvents, brief exposure to high temperatures and prolonged exposure to indefinitely low temperatures. When environmental conditions are adequate, tuns rehydrate and the animals resume metabolic activity. Drying of cells generally leads to massive damage to cellular membranes and proteins, which eventually results in cell death and, consequently, in the death of the entire organism.Heat shock proteins and theirmolecular partners are known to play diverse roles, even in unstressed cells, in successful folding, assembly, intracellular localization, secretion, regulation, and degradation of other proteins. In this study, a complementary focus is on tardigrades undergoing stress in nature and on the roles of stress genes of the hsp10, hsp20/30, hsp40, hsp60, hsp70 and hsp90 family in the stress physiology ofwhole organisms in different life history stages. This study is part of the project www.FUNCRYPTA.de, funded by the German Federal Ministry of Education and Research, BMBF (0313838).

Establishment of a Rearing System of the Extremotolerant Tardigrade Ramazzottius varieornatus: A New Model Animal for Astrobiology

Studies on the ability of multicellular organisms to tolerate specific environmental extremes are relatively rare compared to those of unicellular microorganisms in extreme environments. Tardigrades are extremotolerant animals that can enter an ametabolic dry state called anhydrobiosis and have high tolerance to a variety of extreme environmental conditions, particularly while in anhydrobiosis. Although tardigrades have been expected to be a potential model animal for astrobiological studies due to their excellent anhydrobiotic and extremotolerant abilities, few studies of tolerance with cultured tardigrades have been reported, possibly due to the absence of a model species that can be easily maintained under rearing conditions. We report the successful rearing of the herbivorous tardigrade, Ramazzottius varieornatus, by supplying the green alga Chlorella vulgaris as food. The life span was 35 +/- 16.4 d, deposited eggs required 5.7 +/- 1.1 d to hatch, and animals began to deposit eggs 9 d after hatching. The reared individuals of this species had an anhydrobiotic capacity throughout their life cycle in egg, juvenile, and adult stages. Furthermore, the reared adults in an anhydrobiotic state were tolerant of temperatures of 90 degrees C and -196 degrees C, and exposure to 99.8% acetonitrile or irradiation with 4000 Gy (4)He ions. Based on their life history traits and tolerance to extreme stresses, R. varieornatus may be a suitable model for astrobiological studies of multicellular organisms.

The tardigrade Ramazzottius varieornatus as a model of extremotolerant animals

I report the successful culture of the tardigrade Ramazzottius varieornatus by supplying the green alga Chlorella vulgaris as a food. A standard strain named YOKOZUNA-1 has been isolated from single egg of R. varieornatus. The reared animals have anhydrobiotic ability throughout their life cycle in eggs, juveniles, and adults. Body water content of adult R. varieornatus drops from 78.6% wt./wt. in hydrated state to 2.5% wt./wt. in anhydrobiotic state. Reared adults, while in an anhydrobiotic state, are tolerant of exposure to 100°C, 1 GPa, 99.8% acetonitrile, and 5000 Gy of 4He ions. Furthermore, hydrated active adults survive freezing at -2 to -20°C at a cooling rate of 1°C/min. Based on useful culture systems and extraordinary tolerance to extreme environmental conditions of R. varieornatus, this species is expected to be a suitable model for extremophile research focusing on multicellular organisms.

Nematode communities in the artificially vegetated belt with or without irrigation in the Tengger Desert, China

Shifting sand dunes threatened the Baotou-Lanzhou railway being kept on operation smoothly seriously crossing Shapotou on the southeastern edge of the Tengger Desert (China). Artificial vegetation input was employed in Shapotou and a vegetated belt was established for stabilizing the shifting sand dunes. Nematode communities from bare soil between vegetation (ST, STI) and soil under the vegetation (ST-V, STI-V) in the vegetation belt, were investigated. All sites received natural rainfall, of which STI and STI-V received irrigation supplements. Total 43 genera were found in four types of soil samples, and Acrobeles, Acrobeloides, Chiloplacus, Panagrolaimus, Aphelenchoides and Ditylenchus were dominant genera in our study. Nematode abundance per 100 g fresh soil (47–552), the number of taxa identified (8–20), species richness (1.51–3.41), the proportion of plant feeders (0.3%–4.2%) and abundance of all cp groups responded to vegetation. The proportion of omnivores (0.5%–7%), the number of taxa identified, Shannon index (1.26–2.12), species richness, genus dominance (0.18–0.40) and abundance of cp3–5 responded to irrigation supplements. The application of PCA helped to reveal that almost all nematode taxa exhibited a positive loading on the horizontal axis, it implied that nematode abundance was generally higher on sites with planted input. Besides, it showed a vegetation gradient from bare soil between vegetation to soil under vegetation along the first axis, while an irrigation gradient along the second axis. Similarly, classification analysis based on all cased involved indicated that two-main group of nematodes was distinguished by their habitats under vegetation from bare soil. Those results suggested that vegetation input might play a more important role on nematode community composition than irrigation supplements for stabilization of shifting sand dunes. Therefore, nematode community composition could be as an indicator of stabilization of sand dune approaches of plant input with only natural rainfall versus with natural rainfall in combination with irrigation supplements in our system. However, the apparent anhydrobiotic state of the nematode fauna required special consideration in desert system, thus the utilization of the nematodes as an indicator of stabilization of sand dune regimes should be further explored on anhydrobiotic nematodes.

Volume and morphology changes of a bdelloid rotifer species (Macrotrachela quadricornifera) during anhydrobiosis

Following a study on the changes occurring in a bdelloid species (Macrotrachela quadricornifera, Rotifera, Bdelloidea) when entering anhydrobiosis, we investigated the changes in morphology, including weight and volume during the transition from the active hydrated to the dormant anhydrobiotic state by scanning electron microscopy, confocal microscopy and light microscopy. We compared sizes and morphologies of hydrated extended, hydrated contracted and anhydrobiotic specimens. Bdelloid musculature is defined: longitudinal muscles are contracted in the hydrated contracted animal (head and foot are retracted inside the trunk), but appear loose in the anhydrobiotic animal. When anhydrobiotic, M. quadricornifera appears much smaller in size, with a volume reduction of about 60% of the hydrated volume, and its internal organization undergoes remarkable modifications. Internal body cavities, clearly distinguishable in the hydrated extended and contracted specimens, are no longer visible in the anhydrobiotic specimen. Concomitantly, M. quadricornifera loses more than 95% of its weight when anhydrobiotic; this is more than expected from the volume reduction data and could indicate the presence of space-filling molecular species in the dehydrated animal. We estimate that the majority of body mass loss and volume reduction can be ascribed to the water loss from the body cavity during desiccation.

Tardigrades as a Potential Model Organism in Space Research

Exposure of living organisms to open space requires a high level of tolerance to desiccation, cold, and radiation. Among animals, only anhydrobiotic species can fulfill these requirements. The invertebrate phylum Tardigrada includes many anhydrobiotic species, which are adapted to survive in very dry or cold environmental conditions. As a likely by-product of the adaptations for desiccation and freezing, tardigrades also show a very high tolerance to a number of other, unnatural conditions, including exposure to ionizing radiation. This makes tardigrades an interesting candidate for experimental exposure to open space. This paper reviews the tolerances that make tardigrades suitable for astrobiological studies and the reported radiation tolerance in other anhydrobiotic animals. Several studies have shown that tardigrades can survive gamma-irradiation well above 1 kilogray, and desiccated and hydrated (active) tardigrades respond similarly to irradiation. Thus, tolerance is not restricted to the dry anhydrobiotic state, and I discuss the possible involvement of an efficient, but yet undocumented, mechanism for DNA repair. Other anhydrobiotic animals (Artemia, Polypedium), when dessicated, show a higher tolerance to gamma-irradiation than hydrated animals, possibly due to the presence of high levels of the protective disaccharide trehalose in the dry state. Tardigrades and other anhydrobiotic animals provide a unique opportunity to study the effects of space exposure on metabolically inactive but vital metazoans.

Radiation tolerance in the eutardigrade Richtersius coronifer

Purpose: Tardigrades have a reputation of being extremely tolerant to extreme environmental conditions including tolerance to ionizing radiation while in a desiccated, anhydrobiotic state. However, the evidence for radio-tolerance in tardigrades is based on only one previous report, and there is an obvious need for complementary studies. In this paper we report an investigation on radio-tolerance in desiccated and hydrated specimens of the eutardigrade Richtersius coronifer.Materials and methods: Groups of 30 – 50 tardigrades were exposed to γ-radiation at doses between 1.0 – 9.0 (anhydrobiotic animals) or 0.5 – 5.0 (hydrated animals) kGy and the animals were followed until all were dead. Radiation tolerance of both desiccated and hydrated tardigrades was studied.Results: Both desiccated and hydrated animals irradiated with 0.5 and 1 kGy did not deviate in survival from the control groups. Animals from all exposed groups underwent their moulting and egg production cycle, but at decreasing frequency for doses above 1 kGy. No eggs laid by irradiated animals hatched, while eggs laid by controls did so.Conclusion: Our study suggests that radiation tolerance in tardigrades is not due to biochemical protectants connected with the desiccated state. Rather, cryptobiotic tardigrades may rely on efficient mechanisms of DNA repair, the nature of which is currently unknown.

Dehydration-specific induction of hydrophilic protein genes in the anhydrobiotic nematode Aphelenchus avenae.

Some organisms can survive exposure to extreme desiccation by entering a state of suspended animation known as anhydrobiosis. The free-living nematode Aphelenchus avenae can be induced to enter the anhydrobiotic state by exposure to a moderate reduction in relative humidity. During this preconditioning period, the nematode accumulates large amounts of the disaccharide trehalose, which is thought to be necessary, but not sufficient, for successful anhydrobiosis. To identify other adaptations that are required for anhydrobiosis, we developed a novel SL1-based mRNA differential display technique to clone genes that are upregulated by dehydration in A. avenae. Three such genes, Aav-lea-1, Aav-ahn-1, and Aav-glx-1, encode, respectively, a late embryogenesis abundant (LEA) group 3 protein, a novel protein that we named anhydrin, and the antioxidant enzyme glutaredoxin. Strikingly, the predicted LEA and anhydrin proteins are highly hydrophilic and lack significant secondary structure in the hydrated state. The dehydration-induced upregulation of Aav-lea-1 and Aav-ahn-1 was confirmed by Northern hybridization and quantitative PCR experiments. Both genes were also upregulated by an osmotic upshift, but not by cold, heat, or oxidative stress. Experiments to investigate the relationship between mRNA levels and protein expression for these genes are in progress. LEA proteins occur commonly in plants, accumulating during seed maturation and desiccation stress; the presence of a gene encoding an LEA protein in an anhydrobiotic nematode suggests that some mechanisms of coping with water loss are conserved between plants and animals.

Long‐term anhydrobiotic survival in semi‐terrestrial micrometazoans

AbstractThis study represents the first systematic investigation of long‐term anhydrobiotic survival in tardigrades, rotifers and nematodes inhabiting mosses and lichens. Sixty‐three different samples from public and private collections, kept dry for 9–138 years, were examined. Rotifers of the genus Mniobia and the eutardigrade Ramazzottius oberhaeuseri (hatched from eggs) were found alive from one of the samples (9 years old). These observations represent the longest record for rotifers in the anhydrobiotic state. For tardigrades, our results confirm previous reports on the upper limit of anhydrobiotic survival under atmospheric oxygen conditions. This study suggests the possibility that tardigrade eggs are able to withstand longer periods in anhydrobiosis than animals. Some problems related to the evaluation of long‐term anhydrobiotic survival, such as contamination and chemical treatments of samples, are reported. The possible role of the microenvironment in which the anhydrobiotic animals are kept is discussed.

MonitoringPratylenchus thornei densities in SOCL and roots under resistant (Triticum turgidum durum) and susceptible (Triticum aestivum) wheat cultivars

Pratylenchus thornei densities were monitored in field plots of two winter wheat cultivars in a dry farming area of southern Spain. Samples were taken fortnightly during the wheat-growing season and from the following dry fallow. Under bread wheat cv. ‘Yecora’, densities ofP. thornei increased for 5 to 6 months and then were maintained or slightly decreased thereafter, surviving the summer dry fallow in an anhydrobiotic state (78.2% and 85.3% survival in soil and roots, respectively). Under durum wheat cv. ‘Donpedro’, nematode densities decreased over the growing season, although densities within the roots increased during the first 2 months of the wheat-growing period, indicating that nematodes could penetrate the roots of this cultivar but were unable to reproduce. These observations suggest resistance of wheat cv. Donpedro toP. thornei.

The Nature of Selection on Anhydrobiotic Capacityin Tardigrades

Tardigrades are well known for their capacity to enter an anhydrobiotic state, and remain ametabolic for several years. Yet the evolutionary background, and the forces of natural selection that act on anhydrobiotic capacity have remained unexplored. In this paper, I review the basic problem of anhydrobiosis and the phenotypic traits that may be the main targets of natural selection for improved anhydrobiotic capacity. I also make an attempt to put anhydrobiosis into a life history perspective, and discuss this life form in the perspective of energetic and genetic constraints.

Behavioural and physiological responses of infective juveniles of the entomopathogenic nematode Heterorhabditis to desiccation

Infective juveniles (IJs) of entomopathogenic nematodes (EPNs) are susceptible to a wide variety of environmental factors, including desiccation, which limit their usefulness as biocontrol agents. Although EPNs can be subjected to a gradual loss of water in their natural environment they are not full anhydrobiotes, being able to survive only moderate levels of desiccation at high relative humidities (rh). We investigated the desiccation tolerance of IJs of several Heterorhabditisspecies and strains when exposed to fast and slow desiccation regimes. We also investigated the behavioural and biochemical responses of Heterorhabditis IJs when exposed to 98% rh for 4 days. IJs of H. megidis UK211 (but not IJs of H. indica) aggregate into large clumps when desiccated at high rh, but unlike Steinernema spp., neither H. megidis nor H. indica IJs showed any tendency to coil. Preincubation of H. megidis UK211 IJs at high (98%) rh enhances their ability to survive for 150 min at 57% rh. We show that preincubation of H. megidis and H. indica at 98% rh induces the synthesis of glycerol but not of trehalose, whereas identical preincubation conditions do induce trehalose synthesis in Steinernema carpocapsae and Aphelenchus avenae. The biosynthesis of glycerol rather than trehalose by IJs of two species of Heterorhabditis in response to moderate levels of desiccation indicates that Heterorhabditis is unlikely to have the necessary metabolic responses to desiccation required to enable it to enter into a fully anhydrobiotic state.

Desiccation survival of entomopathogenic nematodes of the genusHeterorhabditis

The present study aims at determining the desiccation tolerance of entomopathogenic nematodes from the genusHeterorhabditis recently isolated in Israel. We first verified the most suitable desiccation conditions that lead to induction of the anhydrobiotic state usingHeterorhabditis bacteriophora HP88. After direct exposure of infective juveniles (IJs) to 97% and 93% r.h. for 96 h, the survival rate was > 70%. By contrast, exposing HP88 IJs to 88% and 85% r.h. resulted in poor survival ( 93% > 88% > 85% r.h.) at a 24-h or 72-h interval resulted in enhanced survival (30% survival) of IJs at the final r.h. level compared with IJs which were either directly exposed to 85% r.h. (0% survival) or were preconditioned at the higher r.h. levels prior to exposure to 85% r.h. (15% survival).H. bacteriophora HP88 IJs were able to survive for at least 18 days after preconditioning. At 97% r.h. nematode viability remained stable at 70–85% and at 93% r.h. survival ranged between 37% and 60%. The data indicate that survival is influenced by rate of water removal from the nematode’s body and a minimal relative humidity level (>93%). Substantial differences in survival ability were observed among IJs of 12 new heterorhabditid populations, isolated from different climatic regions in Israel, which were preconditioned at 97% r.h. for 72 h following by an additional 72 h at 93% r.h. Maximum survival was recorded with HIS-19 (64%), moderate (40–55%) survival was observed with seven isolates, includingH. bacteriophora HP88, and five isolates displayed poor (<25%) desiccation tolerance.

Desiccation stress of entomopathogenic nematodes induces the accumulation of a novel heat-stable protein.

The present study describes a novel heat-stable, water-stress-related protein with a molecular mass of 47 kDa (designated Desc47) in the entomopathogenic nematode Steinernemafeltiae (IS-6). The protein was accumulated about 10-fold (from 7.84 +/- 1.85 to 74.09 +/- 4.35 % relative content level [RCL]) in dehydrated clumps of infective juveniles (IJs), which had lost 344.% of their initial water content (from 65.1 +/- 1.7% to 427 +/- 0.72%) in a desiccation-tolerance-inducing treatment (97% relative humidity [RH] for 3 days). The appearance of Desc47 was accompanied by trehalose accumulation (from 300 to 600 mg trehalose/g protein) during the process of inducing the IJs into a quiescent anhydrobiotic state. A second cycle of IJ dehydration did not alter the RCL of Desc47 (79.3% for the first cycle and 73.3% for the second cycle). Desc47 retained its high RCL (69.7%) in rehydrated active IJs for 3 days, reaching 51.2% of its initial RCL only after a week. No homology to other known proteins was found by mass-spectrometry electrospray-ion-trap analysis. However, of the 5 sequences obtained from the 11protein(ranging from to 21 amino acids), the 21-amino-acid peptide N V A S D A V E T V G N A A G Q A G (D/T) A V showed excellent homology (74% identity in 19 amino acids) to the cold-responsive protein COR14b (g6564861) from Triticum aestivum. In the Caenorhabditis elegans predicted proteome database search, the N21 yielded the first-best identity score (59 % identity in 17 amino acids) to the CE-LEA homologue protein (g2353333). In plants, COR and LEA are related proteins, heat-stable, which are expressed in response to both dehydration and cold acclimation. The implication of the involvement of Desc47 and the osmoprotectant trehalose in the desiccation-tolerance mechanisms of S. feltiae is discussed.

The use of anhydrobiosis by soil nematodes in the Antarctic Dry Valleys

Abstract1. Anhydrobiosis is a survival strategy used by soil nematodes that confers protection from environmental stress and is induced by desiccation.2. To study the use of anhydrobiosis by soil nematodes from the Antarctic Dry Valleys, soil was collected from field sites, and nematodes were extracted using an anhydrobiotic extraction technique. Nematodes were found in a coiled form that characterizes the anhydrobiotic state.3. The proportion of nematodes coiled ranged from 30 to 80% and appeared to reach a maximum in soils drier than 2% gravimetric soil moisture content.4. Wetting of dry soils to 12% gravimetric soil moisture content resulted in rapid uncoiling of nearly 100% of nematodes within 6 h.5. In this cold desert ecosystem, long periods of inactivity appear to be an important aspect of soil nematode life history as their activity may be restricted to the period following infrequent snowfall and melt.

Desiccation tolerance of Muellerius capillaris first-stage larvae from Israeli arid and French temperate habitats and their compatibility to the land snail Theba pisana.

The survival capacities of Muellerius capillaris (Nematoda: Protostrongylidae) free-living larval stages (L1) of an Israeli desert isolate (Nubian isolate) (92 mm rain per annum) and a French isolate from a temperate habitat (650 mm rain per annum) were compared under humid and extreme dry conditions. Under the dry conditions (7% relative humidity [r.h.] and 33% r.h. at 23 degrees C), both isolates exhibited the same remarkable survival capacity for periods of 17 and 28 days, respectively (> 92% survival, P > 0.1). However, during and after recovery from the anhydrobiotic state, different behaviour patterns of L1 were observed in each isolate. Under humid conditions (97% r.h.) for 10 days, both isolates exhibited similarly low survival percentages (13.4 +/- 4.6%, S.E., survival for the Nubian isolate, 3 +/- 3% for the French isolate, P > 0.05). In water, the French isolate was less active, survived better (52 +/- 8.4% vs 28.7 +/- 3.7, by day 28, P < 0.05) and retained a better morphological appearance (shrunk less) than the Nubian isolate. Larval development of the French isolate in the land snail Theba pisana was significantly faster than that of the Nubian isolate, reaching a higher proportion of infective stages (L3) on day 36 post-infection (78.6% vs 56% P > 0.05). However, similar infection intensities were recovered in snails infected with either of the isolates (59.8 +/- 10.3, S.E., n = 14, for the French isolate, 53 +/- 6, n = 13, for the Nubian isolate, P < 0.05).

Desiccation of Rotifers (Macrotrachela Quadricornifera): Survival and Reproduction

Eggs and adults of the bdelloid rotifer Macrotrachela quadricornifera aged 3, 5, 8, and 14 d were dried for 4 d, and 5— and 8—d—old animals for 30 d, to determine ability to recover. The highest percentage recovery was obtained with 8—d rotifers stressed for 4 d., while no animals aged 5 d recovered from 30 d of drying. Age—specific survival and fecundity of animals that recovered from drying generally were similar to controls. Only 5—d—old stressed rotifers had significantly decreased fecundity, while 14—d—old stressed animals had shorter life—spans. Age—specific fecundity and survivorship of stressed were also similar to controls. Age effect ability to recover but not the remaining life of the recovered animals. The life cycle seems to follow a fixed program regardless of the time spent in the anhydrobiotic state.

Consumption of nematodes by fungivorous mites, Tyrophagus spp. (Acarina: Astigmata: Acaridae).

Free-living nematodes may be attacked and eaten by soil mites which are normally considered fungivores or saprophages. Three species in the genus Tyrophagus, common inhabitants of grassland soils and also common pests in stored products, museums and laboratories, are predators of nematodes. All active stages of the mites will voraciously consume nematodes. When offered nematodes and a choice of other food (baker's yeast and algae), 11% of the Tyrophagus putrescentiae, 23% of the T. zachvatkini, and 56% of the T. similis tested fed on nematodes. Tyrophagus zechvatkini and similis were reared on a diet consisting entirely of nematodes, and developed at rates similar to a fungal diet and produced viable offspring. Agar cultures of Aphelenchus avenae which were inoculated with five mating pairs of Tyrophagus zachvatkini had populations that were one-third less than mite-free controls. Observations indicate that nematodes may be attacked by tyrophagid mites when in a dry, anhydrobiotic state.

The effect of dehydration and rehydration on salt loss in the second-stage larvae of Anguina tritici

SUMMARYThe effect of repeated dehydration and hydration on the salt content of 2nd-stage larvae of Anguina tritici was studied. The sodium, potassium, magnesium and calcium contents of the larvae after subjection to varying periods of desiccation and rehydration were determined with atomic absorption spectrophotometry and flame photometry. External cation concentrations on the cuticle of the head, middle body and tail regions were determined semi-quantitatively with scanning electron microscopy in conjunction with X-ray analysis (EDAX). Salt concentrations decreased with each dehydration/rehydration cycle. The greatest loss occurred on revival from the first desiccation period. The results indicated that A. tritici was incapable of regulating it's internal sodium content during revival, but suggested a limited ability to control potassium, magnesium and calcium loss. Salt loss through the nematode cuticle was restricted to potassium and calcium during desiccation. The salt losses encountered are discussed in relation to nematode osmoregulation and to the survival of the nematode in the anhydrobiotic state.