Ancient Adaptive Lateral Gene Transfers in the Symbiotic Opalina-Blastocystis Stramenopile Lineage.

Lateral gene transfer is a very common process in bacterial and archaeal evolution, playing an important role in the adaptation to new environments. In eukaryotes, its role and frequency remain highly debated, although recent research supports that gene transfer from bacteria to diverse eukaryotes may be much more common than previously appreciated. However, most of this research focused on animals and the true phylogenetic and functional impact of bacterial genes in less-studied microbial eukaryotic groups remains largely unknown. Here, we have analyzed transcriptome data from the deep-branching stramenopile Opalinidae, common members of frog gut microbiomes, and distantly related to the well-known genus Blastocystis. Phylogenetic analyses suggest the early acquisition of several bacterial genes in a common ancestor of both lineages. Those lateral gene transfers most likely facilitated the adaptation of the free-living ancestor of the Opalinidae-Blastocystis symbiotic group to new niches in the oxygen-depleted animal gut environment.

Dual role of the Anopheles coluzzii Venus Kinase Receptor in both larval growth and immunity.

Vector-borne diseases and especially malaria are responsible for more than half million deaths annually. The increase of insecticide resistance in wild populations of Anopheles malaria vectors emphasises the need for novel vector control strategies as well as for identifying novel vector targets. Venus kinase receptors (VKRs) constitute a Receptor Tyrosine Kinase (RTK) family only found in invertebrates. In this study we functionally characterized Anopheles VKR in the Gambiae complex member, Anopheles coluzzii. Results showed that Anopheles VKR can be activated by L-amino acids, with L-arginine as the most potent agonist. VKR was not required for the fecundity of A. coluzzii, in contrast to reports from other insects, but VKR function is required in both Anopheles males and females for development of larval progeny. Anopheles VKR function is also required for protection against infection by Plasmodium parasites, thus identifying a novel linkage between reproduction and immunity in Anopheles. The insect specificity of VKRs as well as the essential function for reproduction and immunity suggest that Anopheles VKR could be a potentially druggable target for novel vector control strategies.

Functional analysis of novel aquaporins from Fasciola gigantica.

Fascioliasis, caused by liver flukes of the genus Fasciola, is an important disease of ruminants. In order to identify a potential new drug target we have studied aquaporin (AQP) in Fasciola gigantica. AQPs facilitate the transport of water, glycerol and other small solutes across biological membranes. The structure, function, and pathology of AQPs have been extensively studied in mammals but data for AQPs from trematodes is still limited. In the present study, we have functionally characterized two closely related AQP isoforms, FgAQP-1 and FgAQP-2, from the trematode F. gigantica. Immunohistochemical analysis located the FgAQPs in the tegumental cells, their processes and the tegument itself. In addition, they were present in the epithelial linings of testes and ovary. Expression in Xenopus oocytes of these FgAQPs increased osmotic water permeability 3-4-fold but failed to increase glycerol and urea permeability. AQPs have two highly conserved NPA motifs that are important for the function of the channel pore. In FgAQP-1 and FgAQP-2 the first NPA motif is changed to TAA. Substitution of Thr with Asn in the TAA motif of FgAQP-1 increased its water permeability twofold but did not affect urea and glycerol impermeability while the substitution at the pore mouth of Cys204 by Tyr caused loss of water permeability. In addition, the FgAQPs did not increase methylamine and ammonia permeability after expression in yeast. In comparison to rat AQP-1 the described FgAQPs showed low water permeability and further in vivo analyses are necessary to determine their contribution to osmoregulation in Fasciola.

Evolutionary diversity in polystomatids infecting tetraploid and octoploid Xenopus in East African highlands: biological and molecular evidence.

Species of Protopolystoma are monogenean flukes that only infect allopolyploid hosts in the anuran genus Xenopus. Multivariate analyses of morphometric sclerite characters in the nominal species Protopolystoma simplicis suggest that morphologically distinguishable populations occur in the tetraploid host, Xenopus laevis victorianus, and in each of the octoploid hosts, X. vestitus and X. wittei. The species-level divergence of a lineage specific to X. laevis is supported by sequence variation in the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. Protopolystoma simplicis from X. laevis is redesignated P. microsclera n. sp., with P. simplicis being retained for populations in octoploid hosts. This division is consistent with large differences in egg hatching schedule, fixed differences at the mannose-6-phosphate isomerase and fumarate hydratase loci, and host-specificity in experimental analyses. Although the respective P. simplicis populations in X. vestitus and X. wittei also show significant diversity in allozyme expression, morphometrics and egg hatching schedule, they are retained in the same species because their level of mitochondrial DNA divergence is similar to that found within other Protopolystoma species. The consequences of splitting P. simplicis for a recent interpretation of the origin of Protopolystoma faunas in octoploid Xenopus spp. is discussed.

Postlarval Protopolystoma spp. kidney infections in incompatible Xenopus spp. induce weak resistance to heterospecifics.

Protopolystoma xenopodis and Protopolystoma orientalis are polystomatid monogeneans respectively specific to the parapatric anurans Xenopus laevis and Xenopus muelleri. Parasite larval stages may invade the kidneys of foreign Xenopus spp. but die before migration to the definitive urinary bladder site. Laboratory experiments to assess the effect of a primary incompatible kidney infection on a secondary compatible infection found: (1) a small, significant decrease in the survivorship of P. xenopodis kidney stages (23-37 days p.i. at 25 degrees C) in X. laevis laevis previously challenged with P. orientalis; (2) a significant effect of prior P. orientalis challenge on P. xenopodis development and establishment in the urinary bladder of X. laevis 100 days p.i. (at 21 degrees C); (3) no effect of prior P. xenopodis challenge on adult P. orientalis establishment in X. muelleri (at 21 degrees C), but a significant negative influence on reproductive output (days 0-50 post-patency). Partial cross-resistance to heterospecifics may therefore be induced by Protopolystoma spp. infections in the kidneys of an incompatible host, demonstrating that at least some elements of the host response are non-species specific. The effects observed were weak compared to the strong host resistance known to be generated by an established compatible primary infection with respect to conspecifics. This difference suggests that strong acquired resistance to Protopolystoma species is species-specific and/or induced only by older stages surviving in compatible hosts.

Parasite infectivity to hybridising host species: a link between hybrid resistance and allopolyploid speciation?

Variation in host-specific infectivity was studied in monogenean polystome parasites (Protopolystoma spp.) of the interfertile, parapatric anurans Xenopus laevis laevis and Xenopus muelleri. Laboratory-raised host F1 hybrids were resistant to parasites respectively specific to each parent taxon in nature. This resistance occurred against parasite isolates from both inside and outside a host hybrid/sympatric zone (and no isolate was compatible with the foreign host species under experimental conditions). Geographical Protopolystoma xenopodis isolates showed variable infectivity to a single full-sib group of their usual host, X. l. laevis, and strains with high or low infectivity to these sibs co-occurred in spatially distant local areas (separated by 1,700 km). The host compatibility of P. xenopodis was also subject to host genotypexparasite genotype interactions. Refractoriness to some parasites or pathogens, as a consequence of hybridisation, may have conferred a selective advantage on the allopolyploid pathway by which most Xenopus spp. are believed to have evolved.

Effects of environmental temperature on the susceptibility of Xenopus laevis and X. wittei (Anura) to Protopolystoma xenopodis(Monogenea).

Immune responses in ectothermic vertebrates and the life-processes of their parasites are profoundly linked to ambient temperature, but the functional effect of thermal environment on infectivity and host-specificity in helminths from ectotherms is poorly known. Primary infection establishment of Protopolystoma xenopodis (Monogenea) in compatible hosts ( Xenopus laevis: Anura) is strongly modulated by ambient temperature. Significantly fewer worms survived to the urinary bladder stage at 25 degrees C than at 15 degrees C (relatively high and low temperatures for this system in nature). Pre-infection exposure of X. laevis to cold (10 degrees C) did not significantly affect parasite establishment in the urinary bladder (at 15 or 25 degrees C p.i.), nor shorter-term postlarval survival in the kidneys (at 20 degrees C p.i.), suggesting that residual immunosuppressive effects on host susceptibility are not important. Low temperatures had no permissive effect on the establishment of P. xenopodis in incompatible hosts (Xenopus wittei). The link between thermal conditions and parasitic infection of ectotherms is discussed.

Correlation of parasite speciation and specificity with host evolutionary relationships.

Protopolystoma (Monogenea, Polystomatidae) is strictly specific to the anuran amphibian genus Xenopus. The host group is characterised by a polyploid series in which chromosome numbers reflect diploid, tetraploid, octoploid and dodecaploid constitutions; the series is considered to have evolved through interspecies hybridisation and genome duplication. This study correlates information on host evolutionary relationships with patterns of parasite speciation and host specificity. Protopolystoma is restricted to one subgenus (Xenopus) with multiples of 36 chromosomes, and is absent from the subgenus Silurana (with multiples of 20 chromosomes). Molecular, biochemical and karyotype evidence distinguishes three subgroups within Xenopus. Representative species from each subgroup, Xenopus muelleri, Xenopus fraseri and Xenopus laevis, have been examined for polystomatid infection. Two species of Protopolystoma occur in each of these host species. In X. muelleri, the two Protopolystoma species reflect parasite co-speciation corresponding with the divergence of two sibling host species. Xenopus fraseri and X. laevis (both with 2n = 36 chromosomes) are implicated in the hybrid origin of two octoploid species, Xenopus wittei and Xenopus vestitus (both 2n = 72). The relationships of the Protopolystoma species in these Xenopus taxa reflect this presumed ancestry. Xenopus wittei carries two species of Protopolystoma, one shared with X. fraseri and the other shared with X. laevis. Xenopus vestitus carries a single species of Protopolystoma which is shared with X. laevis but there is no "heirloom" which reflects its hybrid origin involving X. fraseri. In addition to these shared parasite species which may reflect shared host genes, X. fraseri and X. laevis each carry separate species-specific Protopolystoma which do not occur in other Xenopus species even where there is evidence of common genetic information (as in the allopolyploid wittei and vestitus). This case study may be interpreted as indicating a powerful influence of host genetic factors on susceptibility to infection, host-specificity, and parasite speciation.

Reproductive interference in concurrent infections of two Protopolystoma species (Monogenea: Polystomatidae).

The prevention of interspecific reproductive interference is one possible explanation for spatial niche divergence between congeneric monogeneans. However, there is little direct evidence that reproductive interactions with other species are potentially deleterious to the majority of parasitic platyhelminths. Xenopus fraseri-like clawed toads from lowland rainforest in eastern Democratic Republic of Congo are infected by two species of polystomatid monogenean, Protopolystoma fissilis and Protopolystoma ramulosus. Both occur as adults in the host urinary bladder, and exhibit identical copulatory structures and similar body sizes. The small area of the habitat in relation to parasite body size makes close proximity inevitable in concurrent infections. Eggs were collected from five naturally infected hosts: two of these harboured concurrent infections, and three were infected with P. fissilis only. Eggs from concurrent infections showed reduced viability (57.6% embryonation, n = 413) compared with those from P. fissilis-only infections (85.2%, n = 439). This effect may be due to some form of reproductive interference, possibly failure to develop following interspecific cross-fertilisation.

Incompatibility of Protopolystoma xenopodis (Monogenea: Polystomatidae) with an octoploid Xenopus species from southern Rwanda.

Protopolystoma xenopodis is an oviparous monogenean occurring as an adult in the urinary bladder of the clawed toad Xenopus laevis. Oncomiracidia invade the host's kidneys where juveniles develop, subsequently migrating to the definitive site. In central Africa, the tetraploid X. laevis occurs in sympatry with octoploid congeners, including Xenopus wittei, believed to be the hybrid derivatives of X. laevis- and Xenopus fraseri-like parental lineages. Twenty laboratory-raised, naive specimens of an X. wittei-like species from southern Rwanda were each exposed to 30 embryonated P. xenopodis eggs (at 20 degrees C) and screened for parasite egg production until 9 months post-exposure. These toads failed to support the development of gravid parasites (comparable experimental procedures produce at least 35% prevalence of patent infection in the natural host X. laevis). Further X. wittei aff. (n = 26) and X. laevis (n = 17) were exposed to P. xenopodis oncomiracidia and dissected at variable times post-exposure: larvae were able to invade the kidneys of X. wittei aff. and began feeding and morphological development. Severe mortality of juveniles occurred in both natural and unnatural hosts between invasion and 39 days post-exposure. However, while small numbers of parasites persisted in X. laevis, no stages were found in X. wittei aff. beyond 39 days. Present data demonstrate the incompatibility of P. xenopodis with X. wittei aff. and are consistent with a hypothesis that specificity in Protopolystoma-Xenopus systems is determined primarily by the ability of juveniles to complete development in the host's kidneys.

Mutual exclusion of congeneric monogenean species in a space-limited habitat.

Adults of the monogenean genus Protopolystoma infecting Xenopus species occur in an extremely space-limited habitat, the urinary bladder. Xenopus wittei, from a population in Rwanda naturally infected with Protopolystoma fissilis and Protopolystoma simplicis, were exposed to reinfection in captivity (for 1-3 months post-capture) and then monitored in the laboratory for up to 5 months in transmission-free conditions. The two parasites co-occurred in individual bladders less frequently than expected if they were dispersed randomly. Distribution of bladder infections was significantly non-independent (n = 157) and gravid worms of both species were never found in the same host. This pattern might be explained by interference competition between the parasites or by genetic differences in susceptibility within the host species, which is of allopolyploid origin. Other distributional data for sympatric polystomatid species pairs, including P. fissilis and P. ramulosus, show concurrent infections at frequencies consistent with random distributions (i.e. no evidence of interspecific competition or variability in species-specific susceptibility of the hosts). Interference between P. fissilis and P. simplicis (assuming host genetic factors are not involved) may therefore result from a mechanism specific to this species pair. Observations on infection turnover in captive hosts suggest that loss of adult worms may be related to the arrival of juveniles (of either species) in the urinary bladder. Ectopic infection of the host urinary ducts by adult and subadult P. fissilis was observed in some single-species infestations and may be density related. However, the use of an ectopic-site 'refugium' has never been observed in concurrent polystomatid infections.

Diagnosis and treatment of a parasitic dermatitis in a laboratory colony of African clawed frogs (Xenopus laevis).

A parasitic dermatitis, caused by a nematode infestation of the epidermis, was diagnosed in 11 captive African clawed frogs (Xenopus laevis) by a histological examination of the skin. Eight of the animals died or were euthanased before effective treatment with levamisole was administered, and one frog died after being treated. Secondary infection with opportunistic pathogens may have been involved in the progression of the disease.

Infrapopulation dynamics of Gyrdicotylus gallieni (Monogenea: Gyrodactylidae).

Gyrdicotylus gallieni is a viviparous gyrodactylid monogenean that infects the buccal cavity and pharynx of Xenopus laevis laevis. Offspring directly recolonize the same host as their parent and transmission is carried out exclusively by adult worms which are capable of leech-like stepping locomotion. The infrapopulation dynamics (at 20 degrees C) of this species were studied by experimental exposure to single worms of previously uninfected hosts (which were dissected at various time-intervals post-infection). Infrapopulations increased exponentially until 50 days post-infection (p.i.), during which time the intrinsic rate of increase was estimated by regression as 0.065/parasite/day. After this, infrapopulation sizes showed greater variability, with established infections becoming extinct 2-5 months p.i.: elimination was probably due to some form of host reaction, as parasite lineages were maintained for up to 10 months by transfers to successive uninfected hosts. The development of infrapopulations is similar to that reported for other viviparous gyrodactylids parasitic on the body surface of teleosts. However, the intrinsic rate of increase was comparatively slow in G. gallieni and the duration of infection in isolated hosts prolonged. Slow-growing infrapopulations may elicit a host response more slowly and extend the period during which dispersal to a new host or host population can occur. This might be important in G. gallieni for which the internal site of infection could limit the transmission rate, as worm migration from the oral cavity or accidental detachment and expulsion is necessary for host-host transfer to occur.(ABSTRACT TRUNCATED AT 250 WORDS)

Tadpole of the clawed frog, Xenopus laevis, as an experimental intermediate host of Angiostrongylus cantonensis.

Frogs and toads, as paratenic hosts, are known to be sources of human angiostrongyliasis. The present investigation was carried out to examine the role of the tadpole as an experimental intermediate host. First-, second-, and third-stage larvae were found in tadpoles of Xenopus laevis from the 1st, 14th, and 18th days, respectively, after exposure to the first-stage larva of Angiostrongylus cantonensis. Third-stage larvae collected from a frog which had metamorphosed from an exposed tadpole reached maturity and oviposited in an albino rat. This is the first demonstration that a vertebrate, the tadpole of X. laevis, can serve as an experimental intermediate host of A. cantonensis.

[Contribution to the study of Trematodes in Amphibians from the Cameroon (author's transl)]. / Contribution a l'etude des Trématodes d'Amphibiens du Cameroun

Some 800 Amphibians representing 88 species were dissected for parasitic Trematodes of the digestive tract, the lungs and the bladder. 37 species of Anurans (244 specimens) harboured Trematodes. Nine species already found in Africa were observed. A new species, Haematoloechus ocellati n. sp. parasite of the lungs, is described. We propose the transfer of the species Haplometroides eburnense Maeder, 1969 into the genus Plagitura Holl, 1928. A large number of Anurans were examined for the first time in order to study Helminthes. This work brings in the new elements in the biogeography of african Trematodes Amphibians and increases the number of hosts for already mentioned species.