The taxonomic status,host range and geographical distribution of Dollfuschella Vercammen-Grandjean, 1960 (Digenea: Halipeginae) from Xenopus spp. (Anura: Pipidae)

The taxonomy, host range and geographical distribution of halipegine digeneans from Xenopus spp. are reviewed. Dollfuschella Vercammen-Grandjean, 1960 is reinstated from synonymy with Halipegus Looss, 1899 because of the presence of a sinus-sac and weakly developed permanent sinus-organ. Only one representative of the genus, D. rodhaini Vercammen-Grandjean, 1960, is recognised, of which H. rhodesiensis Beverley-Burton, 1963 is considered a synonym. Based on previous literature records this parasite occurs in Xenopus laevis laevis from South Africa and Zimbabwe, X. l. victorianus from Zaire and Uganda, X. l. bunyoniensis from Rwanda, X. wittei from Zaire and Uganda, X. vestitus from Uganda and X. muelleri from Zaire (in some cases host identification has been revised). New host and/or locality records are from X. l. poweri in Zambia, X. l. victorianus in Rwanda, X. l. bunyoniensis in Uganda and X. clivii in Ethiopia. All known hosts belong to a clade characterised by multiples of 2n = 36 chromosomes. There are no records of halipegines from the other major lineage within Xenopus, X. tropicalis-like species with multiples of 2n = 20 chromosomes. This latter group occurs in lowland tropical rain forest from west Africa in contrast to the hosts of D. rodhaini which are found typically in grassland and wooded savanna and in montane forest biotypes. The distribution of D. rodhaini might, therefore, be limited by phylogenetic specificity to the definitive host group or by other ecological factors (e.g., availability of suitable molluscan hosts). Its wide geographical and host range, in common with some other parasite species from the 2n = 36 Xenopus lineage, may result from the lack of ecological or geographical barriers between different definitive host species and subspecies. However, significant geographical variation in egg-size occurs between northern D. rodhaini populations (north of about 15° S) and those from X. l. laevis in southern Africa. This is not considered sufficient for taxonomic recognition but it could reflect the operation of some isolating factor: parasite divergence concurs with evidence that X. l. laevis is evolutionarily relatively distant from the other (more northerly) members of the laevis Rassenkreis.     

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.     

Paramphistome digeneans from shape Xenopus species (Pipidae) in Africa: taxonomy,host-specificity and biogeography

The taxonomy, host range and geographical distribution of paramphistome digeneans from Xenopus spp. in sub-Saharan Africa are reviewed. Two representatives of Progonimodiscus Vercammen-Grandjean, 1960 are recognised, both of which are narrowly or primarily specific to Xenopus. An analysis of morphometric and meristic characters indicated geographical variation in Progonimodiscus doyeri (Ortlepp, 1926), with two allopatric forms showing significant, but continuous, variation in testis size and vitelline follicle number. P. colubrifer n. sp. is distinguished from P. doyeri by the form of muscular elevations on the acetabulum accessory peduncle. It infects Xenopus ( Silurana) tropicalis-like toads from lowland tropical rain forest zones in Nigeria, Togo and the Ivory Coast, while P. doyeri occurs in hosts of the subgenus Xenopus from a wide variety of biotypes. Previous literature records indicate the presence of the southern P. doyeri morphological variant in X. laevis laevis in South Africa and Zimbabwe and the northern variant in X. l. victorianus, X. fraseri aff. and X. muelleri in the Democratic Republic of Congo, X. wittei in Uganda and the Democratic Republic of Congo, and the ranid Conraua crassipes in Cameroon (the only record of Progonimodiscus from a non-pipid host). New host and/or geographical records for this species are of the northern form in X. l. victorianus, X. l. bunyoniensis and X. vestitus in Uganda, X. l. sudanensis in Cameroon, X. borealis in Kenya, X. pygmaeus in the Democratic Republic of Congo, X. fraseri aff. in Cameroon, X. wittei aff. in the Democratic Republic of Congo and X. muelleri in Cameroon and Nigeria. While the geographical limits of the two P. doyeri variants are not known with precision, existing data are consistent with a turnover in the region of 15°S, where a notable discontinuity occurs in the distributions of other Xenopus parasites. Species of Diplodiscus Diesing, 1836 in Xenopus hosts are rare. Diplodiscus peregrinator n. sp. was recovered from X. tropicalis at a single locality in the Ivory Coast and distinguished by a combination of body size, egg size, genital pore position and acetabulum morphology. D. fischthalicus Meskal, 1970 was not found during the present study.     

Studies on the ecology and systematics of a new species of clawed toad, the genus Xenopus, from western Uganda

A new species of African clawed toad, Xenopus kigesiensis sp.n., has been recorded from Lake Mutanda in South-west Kigezi, Uganda. Information on the biology and morphological variation of the species has been compiled from a sample preserved in the field and from a collection of living specimens maintained in the laboratory. An analysis of the characters of the species has been conducted and its relationships with the six known species of the genus assessed.
In the early part of this century the clawed toad populations occupying the lakes of South-West Kigezi exclusively comprised a Xenopus laevis subspecies. It appears that X. kigesiensis sp.n. has recently migrated into Lake Mutanda and has replaced the previous form. Hybridization experiments have demonstrated the reproductive isolation of X. kigesiensis sp.n. from two subspecies of X. laevis. There is evidence of a change in the ecology of the lakes during the period of apparent invasion.
The locality of X. kigesiensis sp.n. lies close to one of the main watersheds of Central Africa; taxonomically the toad forms an important link in the series of known Xenopus species.     

The biology and systematics of a new species of Xenopus (Anura: Pipidae) from the highlands of Central Africa

A new species of clawed toad, Xenopus wittei sp. nov. is described from highland areas overlapping the borders of Rwanda, Uganda and Zaire. A comprehensive morphological analysis of the species has been carried out. X. wittei sp. nov. is very closely related to X. vestitus and both have a tetraploid chromosome complement (2 n = 72), the only examples within the genus. Nevertheless, the two species may be distinguished by easily observed features, including head morphology and dorsal body colours and patterns, and are also identified by their karyotypes and mating calls.
Data on distribution, ecology and parasite infection are derived from both museum collections and fieldwork in Central Africa. X. wittei sp. nov. is sympatric in parts of its range with X. vestitus, X. laevis victorianus and X. I. bunyoniensis , with up to three of the taxa sharing the same habitats. The implications of potential species interactions are discussed. Experimental hybridization has confirmed the reproductive isolation of the two tetraploid species. Parasitological studies show that they may be distinguished from one another by host specific species of the nematode Chitwoodchabaudia and from other Xenopus species by the absence of the tapeworm genus Cephalochlamys . Large scale population movements of the Xenopus species have apparently occurred in the Central African highlands during the period of available records. Sympatry observed amongst the present day representatives of Xenopus may have been an important factor in the evolution of these toads since X. wittei sp. nov. and X. vestitus are interpreted as allopolyploids, the products of interspecies hybridization.     

Differentiation of two locally sympatric Protopolystoma (Monogenea: Polystomatidae) species by temperature-dependent larval development and survival

The developmental response of egg stages to different environmental temperature regimes was studied in Protopolystoma xenopodis and Protopolystoma orientalis (Monogenea: Polystomatidae) isolates from southern Africa. Eggs failed to develop at 10 degrees C, whilst at 15 degrees C only P. xenopodis completed larval development, hatching 49--88 days post-collection. Respective hatching windows were 26--34 (P. xenopodis) and 37--49 (P. orientalis) days at 20 degrees C, and 18--26 and 27--37 days at 25 degrees C. Continuous maintenance at 30 degrees C was lethal for eggs of both species. There were no consistent interspecific differences in the response of egg stages to low and high temperature shocks during early embryonic development.     

Host factors limiting monogenean infections: a case study

Comprehensive field data on polystomatid monogeneans record low prevalence and intensity of infection and suggest that worm burdens in this group are strongly regulated: thus, in the majority of Polystoma species infecting anuran amphibians mean abundance is typically less than one parasite/host. There is circumstantial evidence that the dominant control is attributable to host factors which over-ride variations in transmission success. This review provides a brief summary of information on Pseudodiplorchis americanus, a parasite of the desert toad, Scaphiopus couchii, and then focuses in detail on the spectrum of factors regulating infrapopulations of Protopolystoma xenopodis, a parasite of the aquatic Xenopus laevis. Infection levels of adult worms and their contribution to transmission are regulated by external environmental factors (especially temperature), by host factors (including behaviour and population density), and by a range of parasite factors including intra- and inter-specific competitive interactions and variations in intrinsic characters, especially survivorship and reproductive output. In addition to these factors whose primary effect is to modulate transmission rates, there is a major attrition in parasite numbers between invasion and maturity (3 months post-infection). Long-term laboratory experiments on the Xenopus laevis/Protopolystoma xenopodis interaction demonstrate a powerful acquired immune response. Primary infection is characterised by a high prevalence of established adult worms but the success of subsequent challenge infection is greatly reduced, leading to low prevalence and extended pre-patent period. In the small proportion of hosts supporting a second infection of adult parasites, surviving burdens are small (one to two worms/host) and show reduced egg production. These results provide an explanation for the low burdens encountered in field studies: a majority of adult X. laevis in natural populations are likely to exhibit strong, relatively long-term, post-infection immunity after the loss of a previous infection.     

The mating calls of 12 species and sub-species of the genus Xenopus (Amphibia : Anura)

The principal acoustic characteristics of the mating calls of 12 species and sub-species of the genus Xenopus were determined: X. laevis laevis, X. laevis petersi, X. laevis victorianus, X. gilli, X. muelleri, X. borealis, X. clivii, X. fraseri, X. ruwenzoriensis, X. wittei, X. vestitus and X. tropicalis . These calls are very specific, especially among species whose hybrids are particularly viable in the laboratory. Twin species have not been discovered. Most of the species emit calls with high harmonic frequencies of 16 kHz; these frequencies reach 80 and 150 kHz in X. I. laevis and X. ruwenzoriensis , the ultra-sound level. This was a previously unknown phenomenon in the Batrachians.     

The genusOligolecithus Vercammen-Grandjean (Digenea: Telorchiidae) fromXenopus spp. (Anura: Pipidae), with a description ofO. siluranae n. sp. fromX. tropicalis (Gray) in Ghana

The taxonomy, host range and geographical distribution ofOligolecithus Vercammen-Grandjean, 1960, a genus of telorchiid digeneans fromXenopus spp. in Africa, is reviewed.O. jonkershoekensis Pritchard, 1964 is established as a junior synonym ofO. elianae Vercammen-Grandjean, 1960, andO. siluranae n. sp. is described fromX. tropicalis in Ghana. These two species are differentiated by variations in body length, testicular arrangement and ventral sucker width in proportion to body size.O. elianae occurs inX. laevis laevis from South Africa and Zimbabwe (new locality record),X. l. poweri from Zaire,X. l. victorianus from Zaire, Uganda and Rwanda (new locality record),X. l. bunyoniensis (new host record) from Uganda andX. l. sudanensis from Sudan (new host and locality record). It is also found inX. wittei from Uganda and Zaire andX. vestitus (new host record) from Uganda. The host ofO. siluranae belongs to a separate species group within the genusXenopus from the hosts ofO. elianae; this tropicalis group is phylogenetically isolated, but it also occurs in a different biotype, lowland tropical rain forest, ecologically distinct from the other known host species.     

A redescription of Chabaudus leberrei (Bain & Philippon, 1969) (Nematoda: Seuratoidea) from Xenopus spp. in Swaziland

Chabaudus leberrei (Bain & Philippon, 1969) is redescribed from the pipid anurans Xenopus muelleri (Peters) and X. laevis laevis (Daudin) (new host records) in northern Swaziland, based on light and scanning electron microscope studies. The six anterior protuberances characteristic of the genus Chabaudus Inglis & Ogden, 1965, are, in C. leberrei, formed by bipartite lamellae associated with the internal margins of the three lips. Intraspecific variation in the number and disposition of male caudal papillae and in the development of the cephalic vesicle is documented.     

The morphology and distribution of Xenopus vestitus (Anura: Pipidae) in Central Africa

Studies on the collections of the Musée Royal de l'Afrique Centrale and the Institut Royal des Sciences Naturelles de Belgique have shown that Xenopus kigesiensis Tinsley, 1973 is synonymous with X. vestitus Laurent, 1972. Previously unrecognized representatives of the species have been found in the museum collections and these considerably enlarge the known geographical distribution of X. vestitus. Samples have been recorded principally from highland areas adjacent to the western Rift in Zaire, Uganda and Ruanda; comprehensive morphological analysis demonstrates relatively limited intraspecific variation throughout the known range.
The distribution of X. vestitus overlaps with that of three other Xenopus taxa: X. laevis victorianus, X. bunyoniensis and a new Xenopus species. There is evidence from a large number of collections from widely separated localities that X. vestitus is commonly sympatric with X. l. victorianus. On the other hand, it is postulated that X. vestitus has recently invaded Lake Mutanda, Uganda, and has replaced the original X. bunyoniensis population during the past 30 years. The museum record provides a potential origin of this invasion: X. vestitus populations are well documented at Rutshuru, only 25 km downstream from Lake Mutanda.
A comparison of the morphological characters of X. vestitus, X. l. victorianus and X. bunyoniensis is presented to provide information on the systematic relationships and possible ecological adaptations of the overlapping species.     

Albumin evolution in polyploid species of the genusXenopus

Electrophoresis of serum from 21 Xenopus species and subspecies reveals variable numbers of albumin bands. The diploid X. tropicalis has one albumin, while the tetraploid species (laevis, borealis, muelleri, clivii, fraseri, epitropicalis) have two. The octoploid species (amieti, boumbaensis, wittei, vestitus, andrei) have two to three bands, and the dodecaploid X. ruwenzoriensis has three. The molecular weight of the Xenopus albumins varies from 68 kd (in the tropicalis group) to 74 kd. The subspecies of X. laevis possess two albumins of different molecular weights (70 and 74 kd), whereas most species have only 70-kd albumins. Peptide maps have been obtained from albumin electromorphs by limited proteolysis in sodium dodecyl sulfate (SDS) gels, using S. aureus V8 protease. The peptide patterns produced by electromorphs from the same tetraploid Xenopus species generally differ from each other, suggesting that the two albumin genes contain a substantial amount of structural differences. In addition, the peptide maps are diagnostic for most tetraploid species and for some subspecies of X. laevis as well. Proteolysis of albumins from most octoploid and dodecaploid species results in patterns which are very similar to the ones produced by the electromorphs from X. fraseri. The albumins of X. vestitus differ from those of the other octoploid species. X. andrei possesses two fraseri-type and one vestitus-type albumin, which indicates that it probably originated by allopolyploidy.     

Density dependence of postlarval survivorship in primary infections of Protopolystoma xenopodis

Naive, full-sibling juveniles of Xenopus laevis laevis were exposed to 10, 30, or 50 larvae of the monogenean Protopolystoma xenopodis from an isolate known to produce relatively low adult establishment (typically less than 25% prevalence) in hosts of the same pedigree. Postlarval survival (worms per host) 7 days postinfection (PI), timing from the end of a 10-day infection window, was significantly related and proportional (approximately 31%) to infection dose. Establishment of newly mature adults 90 days PI was low (0-3 worms/host) and unrelated to infection dose. Results confirm that postlarval mortality is severe in primary infections, and that surviving adult infrapopulations appear to be regulated to a very low level (most often 1 worm/ host), even at relatively high larval infection pressures. This density-dependent process could involve direct antagonistic parasite-parasite interactions or indirect interference mediated by parasite-induced host defenses.     

Phylogenetic relationships of the pipid frogs Xenopus and Silurana: an integration of ribosomal DNA and morphology

Relationships of the pipid frog genus Silurana (= Xenopus tropicalis group of some authors) are of particular interest to developmental and molecular biologists because of the purported ancestral (i.e., unduplicated) karyotype of S. tropicalis relative to the genus Xenopus. Although most previous studies have assumed that Silurana is the sister group of Xenopus, recent morphological work suggests that Silurana is more closely related both to the South American genus Pipa and to the African genera Hymenochirus and Pseudhymenochirus than it is to Xenopus. We examined 1,486 bp of relatively variable regions of the ribosomal DNA array (including portions of the 18S and 28S genes, as well as part of an internal transcribed spacer) in Hymenochirus, Silurana, and Xenopus, as well as the outgroup genus Spea, in order to test the alternative hypotheses of relationships for Silurana. Maximum-parsimony analysis using bootstrapping and an analysis using Lake's method of invariants both significantly support the sister-group relationship between Xenopus and Silurana rather than the relationship suggested by morphology. Analysis of the combined morphological/molecular data matrix also significantly supports the Xenopus-Silurana relationship. Although our results are not inconsistent with the recognition of the genus Silurana to accommodate the species formerly called X. tropicalis and X. epitropicalis, the proposed relationships do not require the recognition of this genus in order to render Xenopus monophyletic.     

Evolution of the closely related, sex-related genes DM-W and DMRT1 in African clawed frogs (Xenopus)

DM-W is a dominant, female-specific, regulator of sex determination in the African clawed frog Xenopus laevis. This gene is derived from partial duplication of DMRT1, a male-related autosomal gene. We set out to better understand sex determination in Xenopus by studying this pair of genes. We found that DM-W evolved in Xenopus after divergence from the sister genus Silurana but before divergence of X. laevis and X. clivii, and that DM-W arose from partial duplication of DMRT1β, which is one of the two DMRT1 paralogs in the tetraploid ancestor of Xenopus. Using the rate ratio of nonsynonymous to synonymous substitutions per site and multilocus polymorphism data, we show that DM-W evolved non-neutrally. By cloning paralogs and using a pyrosequencing assay, we also demonstrate that DMRT1 underwent phylogenetically biased pseudogenization after polyploidization, and that expression of this gene is regulated by mechanisms that vary through development. One explanation for these observations is that the expression domain of DMRT1β was marginalized, which would explain why this paralog is dispensable in Xenopus polyploids and why DM-W has a narrow expression domain. These findings illustrate how evolution of the genetic control of stable phenotypes is facilitated by redundancy, degeneration, and compartmentalized regulation.     

Host-defense peptides from skin secretions of the tetraploid frogs Xenopus petersii and Xenopus pygmaeus, and the octoploid frog Xenopus lenduensis (Pipidae)

Peptidomic analysis of norepinephrine-stimulated skin secretions led to the identification of host-defense peptides belonging to the magainin, peptide glycine-leucine-amide (PGLa), and caerulein precursor fragment (CPF) families from the tetraploid frogs, Xenopus petersii (Peters' clawed frog) and Xenopus pygmaeus (Bouchia clawed frog), and the octoploid frog Xenopus lenduensis (Lendu Plateau clawed frog). Xenopsin-precursor fragment (XPF) peptides were not detected. The primary structures of the antimicrobial peptides from X. petersii demonstrate a close, but not conspecific relationship, with Xenopus laevis whereas the X. pygmaeus peptides show appreciable variation from previously characterized orthologs from other Xenopus species. Polyploidization events within the Xenopodinae (Silurana+Xenopus) are associated with extensive gene silencing (nonfunctionization) but unexpectedly the full complement of four PGLa paralogs were isolated from X. lenduendis secretions. Consistent with previous data, the CPF peptides showed the highest growth-inhibitory activity against bacteria with CPF-PG1 (GFGSLLGKALKIGTNLL.NH(2)) from X. pygmaeus combining high antimicrobial potency against Staphylococcus aureus (MIC=6 μM) with relatively low hemolytic activity (LC(50)=145 μM).     

Peptidomic analysis of skin secretions provides insight into the taxonomic status of the African clawed frogs Xenopus victorianus and Xenopus laevis sudanensis (Pipidae)

Peptidomic analysis was used to compare the distribution of host-defense peptides in norepinephrine-stimulated skin secretions from Xenopus victorianus Ahl, 1924 (also described as the subspecies X. laevis victorianus) and Xenopus laevis sudanensis Perret, 1966 with the previously determined distributions in Xenopus laevis (Daudin, 1802) and Xenopus petersii Bocage, 1895. Peptides belonging to the magainin, peptide glycine-leucine-amide (PGLa), and caerulein precursor fragment (CPF) families were purified by reversed-phase HPLC and characterized by electrospray mass spectrometry. Magainin-P2, PGLa-P1, CPF-P1, CPF-P2, and CPF-P3 previously isolated from X. petersii and structurally different from orthologous peptides from X. laevis, were identified in X. victorianus and X. laevis sudanensis skin secretions whereas the corresponding X. laevis peptides were absent. Magainin-1, identical in X. petersii and X. laevis, was also identified in the secretions. Xenopsin-precursor fragment (XPF) peptides, absent from X. petersii but present in X. laevis skin secretions, were not identified in the X. victorianus and X. laevis sudanensis secretions. The data indicate that X. victorianus and X. laevis sudanensis are more closely related to X. petersii than to X. laevis and support separate species status. The study illustrates the value of analysis of host-defense peptides in the evaluation of taxonomic and phylogenetic relationships between closely related frog species.     

Ecological adaptations of grassland‐inhabiting flightless Orthoptera: Fulvoscirtes and Acanthoscirtes,two new genera of African Karniellina (Orthoptera,Tettigoniidae, Conocephalinae,Conocephalini)

Two new genera, Fulvoscirtes n.gen. and Acanthoscirtes n.gen. , are established within the subtribe Karniellina of Conocephalini. Fulvoscirtes is based on Xiphidion kilimandjaricum Sjöstedt, 1909 and Acanthoscirtes on Phlesirtes kevani Chopard from northern Kenya. The majority of Fulvoscirtes spp. are confined to open grasslands in the submontane zone of mountains. Fulvoscirtes contains eight species, seven of which are newly described in this paper. Three species and one subspecies occur on Mt Kilimanjaro. These are F. kilimandjaricum (Sjöstedt) constricted to the southern slopes, F. legumishera n.sp. confined to the northern side and F. sylvaticus n.sp. occurring on the western side of Kilimanjaro and on the eastern slopes of Mt Meru. Fulvoscirtes fulvus n.sp. is divided into two subspecies, F. fulvus fulvus n.ssp. found in the submontane zone of east Kilimanjaro and F. fulvus parensis n.ssp. in submontane to montane localities of the North and South Pare mountains. Fulvoscirtes fulvotaitensis n.sp. occurs in the Taita Hills of southern Kenya. Fulvoscirtes viridis n.sp. is described from savannah habitats between Mts Longido and Meru. Fulvoscirtes laticercus n.sp. is found in the Kenyan highlands, while the most southerly occurring species, Fulvoscirtes manyara n.sp. , is found on Mt Hanang and the Mbulu highlands of northwestern Tanzania. Acanthoscirtes contains three species, of which A. albostriatus n.sp. is described newly from savannah habitas of eastern Kilimanjaro. Information is given on the ecology and the acoustic behaviour of some of the species together with keys to the genera of the Karniellina and the species of Fulvoscirtes and Acanthoscirtes. The genera of Karniellina probably evolved at a time when grasslands spread in East Africa due to an increasing aridification of the climate. The earliest lineage, the genus Karniella, is adapted to more forested habitats while the majority of the genera of Karniellina prefer open grasslands. Major splits within Karniellina probably occurred with the emergence of savannah grasslands due to the ongoing fragmentation of forest habitats several millions years ago, but most species within the genera are geologically young, their radiation being boosted by climatic fluctuations of the past 1–2 Ma.