Transition from sea to land: olfactory function and constraints in the terrestrial hermit crab Coenobita clypeatus

The ability to identify chemical cues in the environment is essential to most animals. Apart from marine larval stages, anomuran land hermit crabs (Coenobita) have evolved different degrees of terrestriality, and thus represent an excellent opportunity to investigate adaptations of the olfactory system needed for a successful transition from aquatic to terrestrial life. Although superb processing capacities of the central olfactory system have been indicated in Coenobita and their olfactory system evidently is functional on land, virtually nothing was known about what type of odourants are detected. Here, we used electroantennogram (EAG) recordings in Coenobita clypeatus and established the olfactory response spectrum. Interestingly, different chemical groups elicited EAG responses of opposite polarity, which also appeared for Coenobita compressus and the closely related marine hermit crab Pagurus bernhardus. Furthermore, in a two-choice bioassay with C. clypeatus, we found that water vapour was critical for natural and synthetic odourants to induce attraction or repulsion. Strikingly, also the physiological response was found much greater at higher humidity in C. clypeatus, whereas no such effect appeared in the terrestrial vinegar fly Drosophila melanogaster. In conclusion, our results reveal that the Coenobita olfactory system is restricted to a limited number of water-soluble odourants, and that high humidity is most critical for its function.     

Tonic immobility in terrestrial isopods: intraspecific and interspecific variability

Many arthropods, including terrestrial isopods, are capable of entering a state of tonic immobility upon a mechanical disturbance. Here we compare the responses to mechanical stimulation in three terrestrial isopods Balloniscus glaber, Balloniscus sellowii and Porcellio dilatatus. We applied three stimuli in a random order and recorded whether each individual was responsive (i.e. showed tonic immobility) or not and the duration of the response. In another trial we related the time needed to elicit tonic immobility and the duration of response of each individual. Balloniscus sellowii was the least responsive species and Porcellio dilatatus was the most, with 23% and 89% of the tested individuals, respectively, being responsive. Smaller Balloniscus sellowii were more responsive than larger individuals. Porcellio dilatatus responded more promptly than the Balloniscus spp. but it showed the shortest response. Neither sex, size nor the type of stimulus explained the variability found in the duration of tonic immobility. These results reveal a large variability in tonic immobility behavior, even between closely related species, which seems to reflect a species-specific response to predators with different foraging modes.     

Life history comparison of two terrestrial isopods in relation to habitat specialization

For many animal species, there is a relationship between life history strategies, as predicted by the r–K-selection theory, degree of habitat specialization and response to habitat alteration and loss. Here we compare two sympatric woodlice species with contrasting patterns of habitat use and geographical distribution. We predict that Atlantoscia floridana (Philosciidae), considered a habitat generalist, would exhibit the r-selected traits, whereas Balloniscus glaber (Balloniscidae), considered a habitat specialist, should have the K-selected traits. We analyzed several life history traits as well as life and fecundity tables using 715 and 842 females of A. floridana and B. glaber, respectively, from populations living in syntopy in southern Brazil. As predicted, most evaluated traits allow A. floridana to be considered an r-strategist and B. glaber a K-strategist: A. floridana showed a shorter lifetime, faster development, earlier reproduction, a smaller parental investment, higher net reproductive rate (R₀), a higher growth rate (r) and a shorter generation time (T) in comparison to B. glaber. A. floridana seems to be a successful colonizer with a high reproductive output. These characteristics explain its local abundance, commonness and wide geographical distribution. On the contrary, B. glaber has a restricted geographical distribution that is mainly associated with Atlantic forest fragments, a biome threatened by deforestation and replacement by monocultures. Its narrow distribution combined with the K-selected traits may confer to this species an increased extinction risk.     

Molecular evolution of the androgenic hormone in terrestrial isopods

In crustaceans, the androgenic gland (AG), thanks to the synthesis of the androgenic gland hormone (AGH), controls the differentiation of the primary and secondary male sexual characters. In this study, we amplified 12 new AGH cDNAs in species belonging to five different families of the infra-order Ligiamorpha of terrestrial isopods. Putative essential amino acids for the production of a functional AGH protein exhibit signatures of negative selection and are strictly conserved including typical proteolytic cleavage motifs, a putative N-linked glycosylation motif on the A chains and the eight Cys positions. An insulin-like growth factor motif was also identified in Armadillidium AGH sequences. The phylogenetic relationships of AGH sequences allowed one to distinguish two main clades, corresponding to members of the Armadillidiidae and the Porcellionidae families which are congruent with the narrow specificity of AG heterospecific grafting. An in-depth understanding of the regulation of AGH expression would help deciphering the interaction between Wolbachia, widespread feminizing endosymbiotic bacteria in isopods, and the sex differentiation of their hosts.     

Widespread fluorescence in terrestrial and marine samples investigated from India

The discovery of green fluorescent protein (GFP) from jellyfish Aequorea victoria has provoked numerous studies to unveil the myriads of biomedical applications. Consequently, several studies also revealed the prevalence of fluorescence in different marine and terrestrial organisms. However, since GFP's discovery or the Nobel prize award on GFP, the fluorescence has not been explored so far from India. The current study presents the widespread fluorescent organisms resources available in India for biomedical and toxicological applications. Fluorescence emission from different plant and animal components were examined by direct observations using UV torchlight. Investigation revealed that blue light excited fluorescence in several organisms. For the first time, this study observed GFP like fluorescence in many terretrial and marine organisms of India. Observations are indicating that fluorescent proteins have essential ecological functions that are yet to be determined. The examined plant and animal components were not bioluminescent. In comparison, the potential untouched research areas on fluorescence aspects are detailed. A thorough review of fluorescent organisms reported hitherto is also provided to spread the current knowledge on fluorescence.     

PAH biotransformation in terrestrial invertebrates--a new phase II metabolite in isopods and springtails

Soil-living invertebrates are exposed to high concentrations of contaminants accumulating in dead organic matter, such as polycyclic aromatic hydrocarbons (PAHs). The capacity for PAH biotransformation is not equally developed in all invertebrates. In this paper, we compare three species of invertebrates, Porcellio scaber (Isopoda), Eisenia andrei (Lumbricidae) and Folsomia candida (Collembola), for the metabolites formed upon exposure to pyrene. Metabolic products of pyrene biotransformation in extracts from whole animals or isopod hepatopancreas were compared to those found in fish bile (flounder and plaice). An optimized HPLC method was used with fluorescence detection; excitation/emission spectra were compared to reference samples of 1-hydroxypyrene and enzymatically synthesized conjugates. Enzymatic hydrolysis after fractionation was used to demonstrate that the conjugates originated from 1-hydroxypyrene. All three invertebrates were able to oxidize pyrene to 1-hydroxypyrene, however, isopods and collembolans stood out as more efficient metabolizers compared to earthworms. In contrast to fish, none of the invertebrates produced pyrene-1-glucuronide as a phase II conjugate. Both Collembola and Isopoda produced significant amounts of pyrene-1-glucoside, whereas isopods also produced pyrene-1-sulfate. A third, previously unknown, conjugate was found in both isopods and springtails, and was analysed further using electrospray and atmospheric pressure chemical ionisation mass spectrometry. Based on the obtained mass spectra, a new conjugate is proposed: pyrene-1-O-(6"-O-malonyl)glucoside. The use of glucose-malonate as a conjugant in animal phase II biotransformation has not been described before, but is understandable in the microenvironment of soil-living invertebrates. In the earthworm, three other pyrene metabolites were observed, none of which was shared with the arthropods, although two were conjugates of 1-hydroxypyrene. Our study illustrates the great variety of the still unexplored metabolic diversity of invertebrate xenobiotic metabolism.     

Immunological relationships of glutamine synthetases from marine and terrestrial enterobacteria

Glutamine synthetase (GS) fromBeneckea alginolytica strain 90 was purified to homogeneity as indicated by sodium dodecyl sulface polyacrylamide gel electrophoresis. The purified enzyme was injected into rabbits, and the antiserum, as well as previously prepared antiserum against the GS fromEscherichia coli, was used in Ouchterlony double diffusion experiments. From the results of these studies, the marine and terrestrial enterobacteria could be separated into five major groups consisting of (i)Beneckea and related organisms, (ii)Photobacterium phosphoreum, P. leiognathi, andP. angustum, (iii) species ofAeromonas, (iv)Xenorhabdus luminescens, and (v) species of terrestrial enterobacteria. Evidence is presented that inB. alginolytica, as in other Gramnegative eubacteria, GS activity is regulated by adenylylation and deadenylylation.     

Effects of microclimate on behavioural and life history traits of terrestrial isopods: implications for responses to climate change

The sensitivity of terrestrial isopods to changes in both temperature and moisture make them suitable models for examining possible responses of arthropod macro-decomposers to predicted climate change. Effects of changes in both temperature and relative humidity on aggregation, growth and survivorship of species of isopods contrasting in their morphological and physiological adaptations to moisture stress have been investigated in laboratory microcosms.All three traits were more sensitive to a reduction in relative humidity of 20–25% than they were to an increase in temperature of 5–6 °C. These results suggest that predicted changes in climate in south east England may reduce the extent to which soil animals stimulate microbial activity and hence carbon dioxide (CO₂) emissions from soils in the future. This may help to mitigate the potential for a positive feedback between increased CO₂ emissions from soils, and increased greenhouse effects causing an increase in soil temperatures.     

Seagrass landscapes: a terrestrial approach to the marine subtidal environment

Subtidal seagrass habitats are prime candidates for the application of principles derived from landscape ecology. Although seagrass systems are relatively simple compared to their terrestrial counterparts in terms of species diversity and structural complexity, seagrasses do display variation in spatial patterns over a variety of scales. The presence of a moving water layer and its influence on faunal dispersal may be a distinguishing feature impacting ecological processes in the subtidal zone. Studying seagrass-dominated landscapes may provide a novel approach to investigating questions regarding self-similarity of spatial patterns, and offers a new perspective for analysing habitat change in a variety of marine environments.     

Operational sex ratio in terrestrial isopods: interaction between potential rate of reproduction and Wolbachia-induced sex ratio distortion

Selfish genetic elements distorting sex ratio are known in several arthropods. By inducing a deficit of males, these sex ratio distorters may modify sexual selection by reversing the sex that competes for a mate. They also have potential to reduce the male proportion to values limiting mating possibilities and therefore limiting population size. Wolbachia endosymbionts are intracytoplasmic, vertically transmitted bacteria that convert genotypic males of terrestrial isopods (woodlice) into functional females. We have tested the impact of these feminizing symbionts on the operational sex ratio (OSR) in three woodlice species. Preliminary experiments consisted in estimating the potential rate of reproduction in males and females, and measuring the dynamics of the onset of reproduction in the wild. These parameters were then combined with population sex ratio to discriminate key factors influencing the OSR. The results suggest that the high potential rate of reproduction of males and the asynchrony in female receptivity both counterbalance female-biased sex ratios. The result is an overall balanced or slightly female-biased OSR. Male deficit can therefore not be considered as a factor strongly limiting reproduction in woodlice. Some females were nevertheless found not mated in the wild at the beginning of the reproductive season, most of them being infected by Wolbachia . This suggests that uninfected females may have an advantage as the first mate. Consequences of these findings on woodlice population dynamics are discussed.     

New primers for the class Actinobacteria: application to marine and terrestrial environments

In this study, we redesigned and evaluated primers for the class Actinobacteria. In silico testing showed that the primers had a perfect match with 82% of genera in the class Actinobacteria, representing a 26-213% improvement over previously reported primers. Only 4% of genera that displayed mismatches did so in the terminal three bases of the 3' end, which is most critical for polymerase chain reaction success. The primers, designated S-C-Act-0235-a-S-20 and S-C-Act-0878-a-A-19, amplified an approximately 640 bp stretch of the 16S rRNA gene from all actinobacteria tested (except Rubrobacter radiotolerans) up to an annealing temperature of 72 degrees C. An Actinobacteria Amplification Resource (http://microbe2.ncl.ac.uk/MMB/AAR.htm) was generated to provide a visual guide to aid the amplification of actinobacterial 16S rDNA. Application of the primers to DNA extracted from marine and terrestrial samples revealed the presence of actinobacteria that have not been described previously. The use of 16S rDNA similarity and DNA-DNA pairing correlations showed that almost every actinomycete clone represented either a new species or a novel genus. The results of this study reinforce the proposition that current culture-based techniques drastically underestimate the diversity of Actinobacteria in the environment and highlight the need to evaluate taxon-specific primers regularly in line with improvements in databases holding 16S rDNA sequences.     

Quantitative estimates of changes in marine and terrestrial primary productivity over the past 300 million years

Changes in marine primary production over geological time have influenced a network of global biogeochemical cycles with corresponding feedbacks on climate. However, these changes continue to remain largely unquantified because of uncertainties in calculating global estimates from sedimentary palaeoproductivity indicators. I therefore describe a new approach to the problem using a mass balance analysis of the stable isotopes (¹⁸O/¹⁶O) of oxygen with modelled O₂ fluxes and isotopic exchanges by terrestrial vegetation for 300, 150, 100 and 50 million years before present, and the treatment of the Earth as a closed system, with respect to the cycling of O₂. Calculated in this way, oceanic net primary productivity was low in the Carboniferous but high (up to four times that of modern oceans) during the Late Jurassic, mid-Cretaceous and early Eocene greenhouse eras with a greater requirement for key nutrients. Such a requirement would be compatible with accelerated rates of continental weathering under the greenhouse conditions of the Mesozoic and early Tertiary. These results indicate possible changes in the strength of a key component of the oceanic carbon (organic and carbonate) pump in the geological past, with a corresponding feedback on atmospheric CO₂ and climate, and provide an improved framework for understanding the role of ocean biota in the evolution of the global biogeochemical cycles of C, N and P.     

Episymbiotic microbes as food and defence for marine isopods: unique symbioses in a hostile environment

Symbioses profoundly affect the diversity of life, often through novel biochemical services that symbionts provide to their hosts. These biochemical services are typically nutritional enhancements and less commonly defensive, but rarely both simultaneously. On the coral reefs of Papua New Guinea, we discovered unique associations between marine isopod crustaceans (Santia spp.) and episymbiotic microbes. Transmission electron microscopy and pigment analyses show that episymbiont biomass is dominated by large (20-30 microm) cyanobacterial cells. The isopods consume these photosymbionts and "cultivate" them by inhabiting exposed sunlit substrates, a behaviour made possible by symbionts' production of a chemical defence that is repulsive to fishes. Molecular phylogenetic analyses demonstrated that the symbiotic microbial communities are diverse and probably dominated in terms of population size by bacteria and small unicellular Synechococcus-type cyanobacteria. Although largely unknown in the oceans, defensive symbioses probably promote marine biodiversity by allowing niche expansions into otherwise hostile environments.     

Microbial characterization of toluene-degrading denitrifying consortia obtained from terrestrial and marine ecosystems

The degradation characteristics of toluene coupled to nitrate reduction were investigated in enrichment culture and the microbial communities of toluene-degrading denitrifying consortia were characterized by denaturing gradient gel electrophoresis (DGGE) technique. Anaerobic nitrate-reducing bacteria were enriched from oil-contaminated soil samples collected from terrestrial (rice field) and marine (tidal flat) ecosystems. Enriched consortia degraded toluene in the presence of nitrate as a terminal electron acceptor. The degradation rate of toluene was affected by the initial substrate concentration and co-existence of other hydrocarbons. The types of toluene-degrading denitrifying consortia depended on the type of ecosystem. The clone RS-7 obtained from the enriched consortium of the rice field was most closely related to a toluene-degrading and denitrifying bacterium, Azoarcus denitrificians (A. tolulyticus sp. nov.). The clone TS-11 detected in the tidal flat enriched consortium was affiliated to Thauera sp. strain S2 (T. aminoaromatica sp. nov.) that was able to degrade toluene under denitrifying conditions. This indicates that environmental factors greatly influence microbial communities obtained from terrestrial (rice field) and marine (tidal flat) ecosystems.     

Isolation and characterization of microsatellite markers from the marine isopods Serolis paradoxa and Septemserolis septemcarinata (Crustacea: Peracarida)

This study reports the successful isolation of highly informative microsatellite marker sets for two marine serolid isopod species. For Serolis paradoxa (Fabricius, 1775), 13, and for Septemserolis septemcarinata (Miers, 1875), eight polymorphic microsatellite markers were isolated using the reporter genome enrichment protocol. The number of alleles per locus (N(A) ) and the observed heterozygosity (H(O) ) encompass a wide range of variation within S. paradoxa (N(A) 3-31, H(O) 6-89%) and S. septemcarinata (N(A) 2-18, H(O) 9-94%). The suitability of the newly isolated markers for population genetic studies is evaluated.     

Carrion cycling in food webs: comparisons among terrestrial and marine ecosystems

In light of current global changes to ecosystem function (e.g. climate change, trophic downgrading, and invasive species), there has been a recent surge of interest in exploring differences in nutrient cycling among ecosystem types. In particular, a growing awareness has emerged concerning the importance of scavenging in food web dynamics, although no studies have focused specifically on exploring differences in carrion consumption between aquatic and terrestrial ecosystems. In this forum we synthesize the scavenging literature to elucidate differences in scavenging dynamics between terrestrial and marine ecosystems, and identify areas where future research is needed to more clearly understand the role of carrion consumption in maintaining ecosystem function within each of these environments. Although scavenging plays a similar functional role in terrestrial and aquatic food webs, here we suggest that several fundamental differences exist in scavenging dynamics among these ecosystem types due to the unique selection pressures imposed by the physical properties of water and air. In particular, the movement of carcasses in marine ecosystems (e.g. wave action, upwelling, and sinking) diffuses biological activity associated with scavenging and decomposition across large, three‐dimensional spatial scales, creating a unique spatial disconnect between the processes of production, scavenging, and decomposition, which in contrast are tightly linked in terrestrial ecosystems. Moreover, the limited role of bacteria and temporal stability of environmental conditions on the sea floor appears to have facilitated the evolution of a much more diverse community of macrofauna that relies on carrion for a higher portion of its nutrient consumption than is present in terrestrial ecosystems. Our observations are further discussed as they pertain to the potential impacts of climate change and trophic downgrading (i.e. removal of apex consumers from ecosystems) on scavenging dynamics within marine and terrestrial ecosystems.     

Life history patterns of terrestrial isopods from mesic habitats in the temperate region of Northern Israel (Isopoda: Porcellionidae,Armadillidae)

The life history patterns of two isopod species from mesic habitats in a temperate region were studied. The isopods: Porcellio chuldaensia (Porcellionidae) and Armadillo sp. ('Brown') (Armadillidae) inhabit heavy soils in northern Israel. Both species breed during spring and early summer, but differ in the number of oocytes, eggs and mancas they produce, and thus in their reproductive allocation. Judging from the structure of their ovaries, they are capable of breeding for a few years. In the laboratory they bred once yearly for two years.     

Laboratory infection of terrestrial isopods (Crustacea: Isopoda) with neoaplectanid and heterorhabditid nematodes (Rhabditida: Nematoda)

Under laboratory conditions, the nematodes Neoaplectana carpocapsae and Heterorhabditis heliothidis were able to infect and develop inside the hemocoel of terrestrial isopods belonging to the genus Porcellio. Armidillidium vulgare was also infected by N. carpocapsae, but this host was less susceptible than Porcellio. Results show that, under suitable ecological conditions, it would be feasible to utilize N. carpocapsae as a biological control agent of sowbugs (Porcellio spp.). This is the first report showing the ability of neoaplectanids and heterorhabditids to invade representatives of the Crustacea.