Determinants of Life-history Evolution in Nematodes

What are the determinants of parasite life-history evolution? Does life-history evolution of parasitic species differ from their free-living relatives? How and why do host and parasite life-history traits covary? Here, Serge Morand and Gabriele Sorci use recent comparative studies to investigate life-history evolution in nematodes which present both parasitic and free-living groups. Application of life-history theory to nematodes suggests that the conventional wisdom concerning the high fecundity of parasitic species could simply be the result of the larger body size of the latter when compared with free-living forms. The authors also emphasize, with the use of examples, that in most cases parasite life-history evolution depends on host features.     

Transgene introgression from genetically modified crops to their wild relatives

Transgenes engineered into annual crops could be unintentionally introduced into the genomes of their free-living wild relatives. The fear is that these transgenes might persist in the environment and have negative ecological consequences. Are some crops or transgenic traits of more concern than others? Are there natural genetic barriers to minimize gene escape? Can the genetic transformation process be exploited to produce new barriers to gene flow? Questions abound, but luckily so do answers.     

The dauer hypothesis and the evolution of parasitism: 20 years on and still going strong

How any complex trait has evolved is a fascinating question, yet the evolution of parasitism among the nematodes is arguably one of the most arresting. How did free-living nematodes cross that seemingly insurmountable evolutionary chasm between soil dwelling and survival inside another organism? Which of the many finely honed responses to the varied and harsh environments of free-living nematodes provided the material upon which natural selection could act? Although several complementary theories explain this phenomenon, I will focus on the dauer hypothesis. The dauer hypothesis posits that the arrested third-stage dauer larvae of free-living nematodes such as Caenorhabditis elegans are, due to their many physiological similarities with infective third-stage larvae of parasitic nematodes, a pre-adaptation to parasitism. If so, then a logical extension of this hypothesis is that the molecular pathways which control entry into and recovery from dauer formation by free-living nematodes in response to environmental cues have been co-opted to control the processes of infective larval arrest and activation in parasitic nematodes. The molecular machinery that controls dauer entry and exit is present in a wide range of parasitic nematodes. However, the developmental outputs of the different pathways are both conserved and divergent, not only between populations of C. elegans or between C. elegans and parasitic nematodes but also between different species of parasitic nematodes. Thus the picture that emerges is more nuanced than originally predicted and may provide insights into the evolution of such an interesting and complex trait.     

Genetic variability in endoparasitic helminths

Biochemical genetic studies have revealed high levels o f polymorphism and heterozygosity in natural populations of endoporasitic helminths. With few exceptions, these parasitic helminths appear just as genetically variable as free-living invertebrates. But this finding is not consistent with the idea that the internal environment o f the host represents a stable and homogeneous habitat which selects for parasite homozygosity. In this article, Steven Nadler examines alternative explanations drawn from the perennial 'selectionist-neutralist' debate-do all genes necessarily confer selective advantages or disadvantages, or are some effectively neutral?     

Purification and characterization of actinogelin, a calcium-sensitive actin-accessory protein, from rat liver

Although cell-free extracts prepared from several types of free-living cells, including Ehrlich tumor cells, macrophages and sea-urchin eggs, readily form gels under low Ca2+ conditions, no such ability to induce actin-related gel has been detected in tissue-cell extracts. Ca2+ -insensitive gelation activity was discovered, however, in several tissue-cell extracts, including liver and brain, provided that the extracts were supplemented with skeletal muscle actin. Based on sodium dodecylsulfate/polyacrylamide gel electrophoretic analysis of the gel, these extracts seem to contain both a Ca2+ -insensitive gelation factor and Ca2+ -sensitive one, actinogelin. A procedure for purification of actinogelin from rat liver was developed, and the properties of actinogelin thus purified were compared with those of Ehrlich tumor cell actinogelin. No appreciable difference was found in these two proteins, and Ca2+ sensitivity (50% inhibition of gelation at 1 microM) was very similar. Some of the molecular characteristics are described, and the importance of the presence of actinogelin in tissue cells is discussed.     

Free-living and aggregate-associated Planctomycetes in the Black Sea

We examined the distribution of uncultured Planctomycetes phylotypes along depth profiles spanning the redox gradient of the Black Sea suboxic zone to gain insight into their respective ecological niches. Planctomycetes phylogeny correlated with depth and chemical profiles, implying similar metabolisms within phylogenetic groups. A suboxic zone sample was split into >?30 and ?30?μm fraction except for members of the Scalindua genus, which were apparently free-living. Sequences from Candidatus Scalindua, known to carry out the anammox process, formed two distinct clusters with nonoverlapping depth ranges. One cluster, only 97.1% similar to the named species, was present at high nitrite/nitrate and low ammonium concentrations in the upper suboxic zone. We propose this sequence type be named 'Candidatus Scalindua richardsii'. A second cluster, containing sequences more similar to 'Candidatus Scalindua sorokinii', was present at high ammonium and low nitrite conditions in the lower suboxic zone. Sequences obtained from the sulfidic zone (1000?m depth) yielded Planctomycetes from two uncharacterized Planctomycetacia clusters and three potentially new genera as well as sequences from the uncultured OP3 phylum.     

Adaptation of ticks to a blood-feeding environment: evolution from a functional perspective

Ticks had to adapt to an existing and complex vertebrate hemostatic system from being free-living scavengers. A large array of anti-hemostatic mechanisms evolved during this process and includes blood coagulation as well as platelet aggregation inhibitors. Several questions regarding tick evolution exist. What was the nature of the ancestral tick? When did ticks evolve blood-feeding capabilities? How did these capabilities evolve? Did host specificity influence the adaptation of ticks to a blood-feeding environment? What are the implications of tick evolution for future research into tick biology and vaccine development? We investigate these questions in the light of recent research into protein superfamilies from tick saliva. Our conclusions are that the main tick families adapted independently to a blood-feeding environment. This is supported by major differences observed in all processes involved with blood-feeding for hard and soft ticks. Gene duplication events played a major role in the evolution of novel protein functions involved in tick-host interactions. This occurred during the late Cretaceous and was stimulated by the radiation of birds and placental mammals, which provided numerous new niches for ticks to adapt to a new lifestyle. Independent adaptation of the main tick families to a blood-feeding environment has several implications for future tick research in terms of tick genome projects and vaccine development.     

Cryptic diversity of free-living parabasalids, Pseudotrichomonas keilini and Lacusteria cypriaca n. g., n. sp., as inferred from small subunit rDNA sequences

Ultrastructural and molecular phylogenetic evidence indicate that the Parabasalia consists of seven main subgroups: the Trichomonadida, Honigbergiellida, Hypotrichomonadida, Tritrichomonadida, Cristamonadida, Spirotrichonymphida, and Trichonymphida. Only five species of free-living parabasalids are known: Monotrichomonas carabina, Ditrichomonas honigbergii, Honigbergiella sp., Tetratrichomonas undula, and Pseudotrichomonas keilini. Phylogenetic analyses show that free-living species do not form a clade and instead branch in several different positions within the context of their parasitic relatives. Because the diversity of free-living parabasalids is poorly understood, the systematics of these lineages is in a significant state of disarray. In order to better understand the phylogenetic distribution of free-living parabasalids, we sequenced the small subunit rDNA from three different strains reminiscent of P. keilini; the strains were isolated from different geographical locations: (1) mangrove sediments in Japan and (2) sediments in Cyprus. These data demonstrated that the free-living parabasalids P. keilini and Lacusteria cypriaca n. g., n. sp., form a paraphyletic assemblage near the origin of a clade consisting mostly of parasitic trichomonadids (e.g. Trichomonas vaginalis). This paraphyletic distribution of similar morphotypes indicates that free-living trichomonadids represent a compelling example of morphostasis that provides insight into the suite of features present in the most recent free-living ancestor of their parasitic relatives.     

The classification of free-living spirochetes

Summary The natural relationships among free-living and host-associated spirochetes were studied by investigating aspects of their morphology, physiology and DNA base composition. These studies indicated that the strictly and facultatively anaerobic, free-living spirochetes share characteristics which distinguish them from their host-associated counterparts and from the free-living leptospires.It is suggested that the strictly and facultatively anaerobic, free-living spirochetes be grouped in the genus Spirochaeta and that this genus include the following four species: S. plicatilis (type species), S. stenostrepta, S. zwelzerae (Treponema zuelzerae), and S. aurantia. The characteristics of these species are described.     

Health of adult free-living cheetahs

Health of free-living adult cheetahs was assessed using haematological and biochemical measurements of condition. Results showed that cheetahs of both sexes varied on several of these measures, demonstrating that their genetic monomorphism does not result in individuals being of equivalent health. Differences in condition appeared to be somewhat associated with sex, age and whether males were territorial. Compared to two different groups of captive cheetahs, free-living cheetahs were monocytotic, and free-living females were macrocytic. Limited data from captive animals showed they ate more than their free-living counterparts, possibly reflecting the elevated blood glucose levels of captive cheetahs.     

Psittacine beak and feather disease in a free-living ring-necked parakeet (Psittacula krameri) in Great Britain

The ring-necked parakeet (RNP), Psittacula krameri, is an invasive species in Great Britain (GB) which is undergoing rapid population expansion in the wild. Although it has been suggested that RNPs could be a potential source of infectious disease, little research has been done on the pathogens infecting this species in GB. Psittacine beak and feather disease (PBFD), caused by beak and feather disease virus (BFDV), is an important infectious disease of psittacines, including captive RNPs, in GB and elsewhere. A wild RNP with marked feather abnormalities observed in an urban garden in London was euthanased and examined post mortem. Plucked contour feathers and pooled liver and spleen were PCR-positive for BFDV DNA. Histopathological examination of affected skin demonstrated BFDV-compatible lesions. A feather from another RNP from a different location also was PCR-positive for BFDV. This is the first report of PBFD in a wild free-living bird in GB. BFDV only affects psittacines; therefore, there is no known risk to native British birds. The presence of BFDV in free-living RNPs, however, could pose a disease threat to captive psittacines. Further work is required to determine the distribution and impact of BFDV infection in free-living RNPs in GB. Whether this case represents sporadic disease associated with established endemic infection or the index case of an emergent disease is currently unknown.     

Mutation rate and genome reduction in endosymbiotic and free-living bacteria

Genome reduction has been considered the hallmark of endosymbiotic bacteria, such as endocellular mutualists or obligatory pathogens until it was found exactly the same in several free-living bacteria. In endosymbiotic bacteria genome reduction is mainly attributed to degenerative processes due to small population size. These cannot affect the free-living bacteria with reduced genomes because they are known to have very large population sizes. It has been proposed that selection for simplification drove genome reduction in these free-living bacteria. For at least one of them (Prochlorococcus), genome reduction is associated with accelerated evolution and we suggest an alternative hypothesis based on increase in mutation rate as the primary cause of genome reduction in free-living bacteria.     

Mediastinal lymphoma in a badger (Meles meles)

A case of mediastinal lymphoma in a free-living, adult male European badger (Meles meles), from Trento Province (northern Italy) is described. A yellow-red mass delimited by a thick fibrous capsule completely occupied the anterior mediastinum. Based on the histologic and immunohistochemical findings, a diagnosis of mediastinal lymphocytic lymphoma was made. Documentation of neoplasms in free-living species is important for comparison with domestic animals and humans and may give insight into epidemiology of these diseases. To the authors' knowledge, this is the first report of a mediastinal lymphoma in a badger.     

Development of microbial biofilms on various surfaces for the treatment of heavy metal containing effluents

Ground glass and roughened polystyrene model stream beds supported mixed microbial biofilms capable of removing Cu , Cd and Pb from running waste waters. Metal uptake by these biofilms was compared to that of similar free-living, mixed microbial populations exposed to different metal concentrations and combinations. Metal uptake by the biofilms was on average 17 times better than that by the free-living cultures.     

Diversity and activity of free-living nitrogen-fixing bacteria and total bacteria in organic and conventionally managed soils

Agricultural soils are heterogeneous environments in which conditions affecting microbial growth and diversity fluctuate widely in space and time. In this study, the molecular ecology of the total bacterial and free-living nitrogen-fixing communities in soils from the Nafferton Factorial Systems Comparison (NFSC) study in northeast England were examined. The field experiment was factorial in design, with organic versus conventional crop rotation, crop protection, and fertility management factors. Soils were sampled on three dates (March, June, and September) in 2007. Total RNA was extracted from all soil samples and reverse transcribed. Denaturing gradient gel electrophoresis (DGGE) and quantitative PCR (qPCR) were used to analyze nifH and 16S rRNA genes in order to study free-living diazotrophs and the total bacterial community, respectively. Crop rotation was shown to have a significant effect on total bacterial diversity (and that of free-living N fixers) (P ≤ 0.001). On all three dates, nifH activity was higher in the conventional crop rotation. In contrast, qPCR analysis of free-living N fixers indicated significantly higher levels of activity in conventionally fertilized plots in June (P = 0.0324) and in plots with organic crop protection in September (P = 0.0143). To our knowledge, the effects of organic and conventional farming systems on free-living diazotrophs have never been studied. An increased understanding of the impacts of management practices on free-living N fixers could allow modifications in soil management practices to optimize the activity of these organisms.     

Evolutionary loss of parasitism by nematodes? Discovery of a free-living filaroid nematode

A cattle-drinking pool in nature reserve "Zwin" on the Belgian coast contained free-living third-stage infective filaroid juveniles. These juveniles clearly differ morphologically from all known nematodes. Morphological and molecular analyses indicate a position within the Filaroidea. The aberrant biology of this nematode, namely, a free-living stage in an aquatic environment, is unknown within this superfamily, and the evolution of the parasitic phenotype to a free-living state is generally thought to be unlikely. However, the obtained placement in the small subunit molecular phylogenetic tree suggests that this free-living stage is most likely a secondary adaptation. It is reasonable to assert that nematodes with complex life cycles still have the genetic potential for a reversion from parasitism to a (partial) free-living stage.     

Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria.

Symbiotically associated cyanobacteria from Azolla mexicana and Azolla pinnata were isolated and cultured in a free-living state. Morphological analyses revealed differences between the free-living isolates and their symbiotic counterparts, as did restriction fragment length polymorphism (RFLP) analyses with both single-copy glnA and rbcS gene probes and a multicopy psbA gene probe. RFLP analyses with Anabaena sp. strain PCC 7120 nifD excision element probes, including an xisA gene probe, detected homologous sequences in DNA extracted from the free-living isolates. Sequences homologous to these probes were not detected in DNA from the symbiotically associated cyanobacteria. These analyses indicated that the isolates were not identical to the major cyanobacterial symbiont species residing in leaf cavities of Azolla spp. Nevertheless, striking similarities between several free-living isolates were observed. In every instance, the isolate from A. pinnata displayed banding patterns virtually identical to those of free-living cultures previously isolated from Azolla caroliniana and Azolla filiculoides. These results suggest the ubiquitous presence of a culturable minor cyanobacterial symbiont in at least three species of Azolla.     

Identification of a common cyanobacterial symbiont associated with Azolla spp. through molecular and morphological characterization of free-living and symbiotic cyanobacteria.

Symbiotically associated cyanobacteria from Azolla mexicana and Azolla pinnata were isolated and cultured in a free-living state. Morphological analyses revealed differences between the free-living isolates and their symbiotic counterparts, as did restriction fragment length polymorphism (RFLP) analyses with both single-copy glnA and rbcS gene probes and a multicopy psbA gene probe. RFLP analyses with Anabaena sp. strain PCC 7120 nifD excision element probes, including an xisA gene probe, detected homologous sequences in DNA extracted from the free-living isolates. Sequences homologous to these probes were not detected in DNA from the symbiotically associated cyanobacteria. These analyses indicated that the isolates were not identical to the major cyanobacterial symbiont species residing in leaf cavities of Azolla spp. Nevertheless, striking similarities between several free-living isolates were observed. In every instance, the isolate from A. pinnata displayed banding patterns virtually identical to those of free-living cultures previously isolated from Azolla caroliniana and Azolla filiculoides. These results suggest the ubiquitous presence of a culturable minor cyanobacterial symbiont in at least three species of Azolla.     

Species-specific differences in heterogonic development of serially transferred free-living generations of Strongyloides planiceps and Strongyloides stercoralis

The direction of free-living development (homogonic vs. heterogonic) in Strongyloides stercoralis and Strongyloides planiceps was examined by successive transplantation of the uterine eggs of free-living females into a test tube culture system containing fresh feces. The eggs from the first-generation free-living females of S. stercoralis did not develop into second-generation free-living adult worms, but all developed into filariform larvae. However, the majority of S. planiceps eggs from the first-generation free-living females developed into second-generation free-living adults. By successive transfer of uterine eggs of each generation, 9 generations developed, and in every cycle more adult worms developed than filariform larvae. However, the number of free-living generations was not infinite; in experiments repeated twice, the number of worms developing from the eggs of eighth or ninth generations was too small to continue further culture. These findings indicate that the pattern of free-living development is different between the 2 species.