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.     

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.     

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 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.     

Speciation of Protopolystoma Bychowsky, 1957 (Monogenea: Polystomatidae) in hosts of the genus shape Xenopus (Anura: Pipidae)

The taxonomy, geographical distribution and hostrange of the polystomatid genus ProtopolystomaBychowsky, 1957 are reviewed. P. xenopodis(Price, 1943) and five new species are recognised,which occur in clawed toads ( Xenopus spp.)throughout subsaharan Africa. Of the two clawed toadsubgenera, Xenopus and Silurana, only theformer is infected. Protopolystoma spp. aredifferentiated by morphological variation of the gut,large hamulus and penis armature. P.xenopodis is found in Xenopus laevis subspeciesin South Africa, Transkei, Zimbabwe, DemocraticRepublic of Congo (D.R.C.), Rwanda, Uganda, Kenya andCameroon ( X. l. poweri and X. l.sudanensis are new host records). It also occurs inintroduced populations of X. l. laevis in theUnited States (southern California) and United Kingdom(South Wales). In subsaharan Africa the speciesdisplays significant, but continuous, geographicalvariation of penis spine size between southernpopulations in X. l. laevis and those in morenortherly host subspecies. Data on the natural hostrange of this parasite were complemented by anexperimental study of host-specificity in the southernform. This can produce patent infections in X.l. victorianus and X. gilli, but not X.wittei nor X. (Silurana) tropicalis. P.simplicis n. sp. is endemic to central and eastAfrican areas, infecting X. laevis subspecies ineastern D.R.C., Rwanda, Uganda and western Kenya, X. wittei-like hosts in eastern D.R.C., westernUganda, Rwanda and Burundi, X. vestitus inwestern Uganda and Xenopus sp. at Nairobi,Kenya. P. ramulosus n. sp. occurs in X.fraseri-like toads in eastern D.R.C. (Gabon andCameroon are also possible literature records), and P. fissilis n. sp. is found in X. fraseri-and X. wittei-like species in Cameroon andeastern D.R.C., and in southern Rwanda, respectively. Two Protopolystoma taxa are found in X.muelleri populations now suspected to representdistinct species: P. occidentalis n. sp. occursin X. muelleri (western form) in Ghana, Togo,Nigeria and Cameroon, while P. orientalis n. sp.is found in X. muelleri (eastern form) in SouthAfrica, Zimbabwe and Tanzania. The allopatricallydistributed species P. ramulosus, P.simplicis, P. occidentalis and P.orientalis form a relatively homogenous grouping withsome interspecific morphological overlap. These taxaare distinguished from P. xenopodis by penisspine morphology and from P. fissilis by hamulusroot form and aspects of gut morphology. Unidentified Protopolystoma sp. have been recorded in X. clivii in Ethiopia, X. fraseri aff. inCameroon and Xenopus sp. in Kenya and Tanzania. At some localities, single host species were infectedby two representatives of Protopolystoma. P. fissilis was recorded in eastern D.R.C. with P. ramulosus, with Protopolystoma sp. inCameroon in X. fraseri-like hosts and with P. simplicis in X. wittei-like hosts in Rwanda. P. xenopodis co-occurred with P. simplicisin X. laevis subspecies through central and eastAfrica.     

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.     

The karyotype of the tetraploid species Xenopus vestitus Laurent (Anura: pipidae)

Xenopus vestitus possesses 72 chromosomes, wherease in the majority of known Xenopus species there are 2n = 36. During meiosis, 36 bivalents are usually observed at metaphase I and 36 chromosomes at metaphase II. Arranged according to size and centromere position, the chromosomes form the same basic morphologic groups typical of the genus Xenopus. However, the groups are composed of quartets of four similar chromosomes instead of diploid pairs of homologs. The exception to this arrangement involves chromosomes bearing secondary constrictions, which in X. vestitus are represented by two different pairs of homologs, one of which shows, in 39% of the observed mitoses, somatic association and is, therefore, considered to carry the nucleolar organizer. X. vestitus represents either a case of ancient autotetraploidy or, more likely, one of allotetraploidy of more modern origin.     

EVOLUTIONARY INFERENCES FROM RESTRICTION MAPS OF MITOCHONDRIAL DNA FROM NINE TAXA OF XENOPUS FROGS

Restriction endonuclease cleavage maps were prepared by the double digestion method for mitochondrial DNAs (mtDNAs) purified from Xenopus borealis, X. clivii, X. fraseri, X. muelleri, X. ruwenzoriensis, X. vestitus, X. laevis victorianus, X. l. laevis, and a variant of X. laevis designated X. laevis “davis.” An average of 21 cleavage sites per genome were mapped with 11 restriction endonucleases. Among the four invariant sites found are three conserved not only among the Xenopus mtDNAs tested but also among nearly all vertebrate mtDNAs examined to date. Two of these are Sac II sites in the 12S and 16S ribosomal RNA genes, and one is a Hpa I site in the gene for asparagine transfer RNA. These three sites permit the alignment and comparison of mtDNAs from different vertebrate classes. Although most of the differences observed among the Xenopus maps are attributable to point mutations causing gain or loss of restriction sites, the maps also differ by three large length mutations in or near the displacement loop. Phylogenetic analysis of 30 informative sites suggests that those members of the laevis species-group that have 36 chromosomes per somatic cell can be divided into three subgroups: 1) X. borealis, X. clivii, and perhaps X. fraseri (the “borealis” subgroup), 2) X. muelleri, and 3) the subspecies of X, laevis. The mtDNA of the hexaploid (2n = 108) species, X. ruwenzoriensis, is most similar to that of taxa in the latter two subgroups, which contrasts with the morphological similarity of this species to X. fraseri. X. ruwenzoriensis may be an allopolyploid with a mother (the contributor of the cytoplasmic mtDNA genome) on the X. laevis or X. muelleri lineage and a father on the X. fraseri lineage. We present a model showing how mtDNA and nuclear genomes can yield contrasting phytogenies for species-groups that have undergone several rounds of interspecific hybridization. Comparison of mitochondrial and nuclear sequence divergences suggests that Xenopus mtDNA, like that of mammals and birds, evolves faster than nuclear DNA. Genetic distances among mtDNAs of Xenopus species are very large, generally approaching or exceeding one substitution per nucleotide.     

ZOOPLANKTON OF LAKES MUTANDA, BUNYONYI AND MULEHE

Quantitative samples of zooplankton from three lakes in the Kigezi District of Uganda have been studied. The systematics of the zooplankton are considered, and some of the identifications given by Worthington & Ricardo (1936) in a previous study of one of these lakes are revised.
Lake Mulehe is the shallowest of the three lakes and contains the largost standing crop of zooplankton. This is in agreement with chemical data which indicato that the supply of nutrient salts in Lake Mulehe is higher than in the other two lakes.
In October 1962 the zooplankton of Lake Mutanda was characterized by the relative abundance of three species of Daphnia which were not found in the samples from the other lakes, although two of these species were present in Lake Bunyonyi in 1931. Rotifers were sparse in Lako Mutands, but here dominant by Tetramastix opoliensis. Lake Bunyonyi was richer in rotifers, but here the dominant species was Keratella tropica , whilo in Lake Mulehe the dominant rotifer was Synchaeta pectinata.
The zooplankton of Lake Mutanda in October 1962 was similar in composition to that of Lake Bunyonyi in 1931, but in 1962 the zooplankton of Lake Bunyonyi was more like that of Lake Mulehe. The possible causos of this change are discussed.     

Where ichthyofaunal provinces meet: the fish fauna of the Lake Edward system,East Africa

Based on literature, museum collections and three recent expeditions, an annotated species list of the Lake Edward, East Africa, drainage system is presented, excluding the endemic haplochromines. A total of 34 non-Haplochromis species belonging to 10 families and 21 genera are recorded from the system. Three of these are endemic and two others have been introduced in the region. Six species are new records for the Lake Edward system. A species accumulation curve indicates that we probably covered most of the non-Haplochromis species in the area sampled during the recent expeditions, but undetected species might still be present in the Congolese part of the system, which is poorly sampled. A comparison of the species list with those of neighbouring basins confirmed the placement of the Lake Edward system within the east-coast ichthyofaunal province.     

Evidence of stenotopy and anthropogenic influence on carbon source for two major riverine fishes of the Lake Victoria watershed

Stable isotope analysis was used to investigate the migratory status and to determine the relative contribution of allochthonous and autochthonous sources of carbon for the major riverine fish species ( Barbus altianalis and Labeo victorianus ) in rivers draining the Kenyan side of Lake Victoria. The two fish species derived carbon from both C4 and C3 plant sources, although L. victorianus exhibited less enriched isotopic carbon values. Fish samples from stations under direct influence of effluents from sugar factories exhibited enriched δ¹³C signals. Assuming that this reflects carbon sourcing from riparian C4 plants, it suggests that carbon from terrestrial sources can be a major energy source in some rivers. This heavy carbon enrichment associated with sugar factories was spatially restricted and occurred in all seasons, implying that sub-populations of the two fish species are non-migratory. The large migratory populations of these two species, for which Lake Victoria was once famous, may be no more.     

The Rift Valley is a major barrier to dispersal of African clawed frogs (Xenopus) in Ethiopia

The Ethiopian highlands – home to striking species diversity and endemism – are bisected by the Rift Valley, a zone of tectonic divergence. Using molecular data we examined the evolutionary history of two co‐distributed species of African clawed frog (Xenopus clivii and X. largeni) that are endemic to this region. Our field collections substantially extend the known distribution of X. largeni, a species formerly known from highlands southeast of the Rift, but that also occurs to the northwest. In both species, analysis of mitochondrial DNA and 19 autosomal loci identifies significant population structure, suggests little or no recent migration across the Rift Valley, and provides divergence time estimates across the Rift of ～1–3.5 million years. These results indicate that the Ethiopian Rift Valley is a major obstacle to dispersal of highland‐adapted amphibians.     

Avian egg collections: museum collection bias driven by shape and size

Avian eggs exhibit substantial intra- and interspecific variation in shape, size and colour. Considerable efforts have been made to better understand the evolutionary drivers behind such variation, often using museum egg collections. Usually it is assumed that museum collections accurately represent the variation seen in natural populations, but this may not be the case if there is collection bias. Collection bias may lead to the over-representation of certain egg traits in collections, due to the aesthetic (or other) preferences of collectors. The aim of this study is to begin to look for the occurrence of potential collection bias in museum egg collections by comparing three shape indices (pointedness/asymmetry, elongation and polar asymmetry) and egg volume between subsets of eggs in museum collections with those of recently sampled eggs in the field for three different bird species: common guillemot Uria aalge, razorbill Alca torda and northern fulmar Fulmarus glacialis. We found no evidence of collection bias in our sampled razorbill and northern fulmar museum collection eggs, but some evidence for a bias in sampled museum collection eggs of common guillemots. Since the guillemot's egg differs from most bird eggs in being pyriform, we suggest that collection bias by historic egg collectors may be more prevalent in species with extreme egg traits. Researchers using museum egg collections to examine questions relating to egg shape should be aware of collection bias risks and consider how to minimise the effect of these possible biases on accumulated datasets.     

The comparative osteology and phylogenetic relationships of African and South American lungfishes (Sarcopterygii: Dipnoi)

Lepidosirenidae is a clade of freshwater lungfishes that include the extant South American Lepidosiren paradoxa Fitzinger, 1837 and African species of the genus Protopterus. These genera have been geographically separated since the break‐up of Gondwana in the Early Cretaceous, but they display similar biology and morphology. Species were distinguished by a combination of features such as head‐to‐body ratios, the number of pairs of ribs, and the presence of external gills, but no discrete skeletal characters were identified, and no comparative studies including all extant species have been published. I used computed tomography (CT), X‐ray photography, and specimens from museum collections to describe the skeletal morphology of all species of lepidosirenid in a comparative context. I digitally disarticulated the bones in each specimen to compile a comparative atlas of the cranial and pectoral elements of all extant lungfishes, which has the potential to increase the correct identifications of specimens in museum collections. The morphology of the frontoparietal, parasphenoid, supraorbital, and suboperculum differ between species. I used those characters, along with molecular sequence data from the ribosomal RNA gene 16S, to run combined morphological and molecular phylogenetic analyses. Lepidosirenidae is monophyletic in all analyses, but the interrelationships of the species of Protopterus vary with the different sources of character data. © 2015 The Linnean Society of London     

Micro‐CT X‐rays do not fragment DNA in preserved bird skins

Most zoological systematics studies are currently based on morphological features, molecular traits or a combination of both to reconstruct animals’ phylogenetic history. Increasingly, morphological studies of museum specimens are using X‐ray computed tomography to visualize internal morphology, because of its ‘non‐destructive’ nature. However, it is not known whether CT can fragment the size of DNA extracted from museum specimens, as has been demonstrated to occur in living cells. This question is of paramount importance for collections based research because X‐rays may reduce the amount of data obtainable from specimens. In our study, we tested whether exposure of museum bird skins to typical CT X‐ray energies (for visualization of the skeleton) increased DNA strand fragmentation, a key factor for the success of downstream molecular applications. For the present study, we extracted DNA from shavings of 24 prepared and dried bird skins (100⁺ years) footpads before and after CT scanning. The pre‐ and post‐CT fragmentation profiles were assessed using a capillary electrophoresis high‐precision instrument (Agilent Bioanalyzer). Comparison of the most common strand length in each DNA sample (relative mass) revealed no significant difference unexposed and exposed tissue (paired t‐test p = 0.463). In conclusion, we found no further quantifiable degradation of DNA strand length under standard X‐ray exposure obtained from our bird skins sample. Differences in museum preservation techniques probably had a greater effect on variation of pre‐CT DNA fragmentation.     

Chromosome complements of the genus Xenopus

The oytogenetic analysis of the genus Xenopus shows that X. laevis laevis, X. laevis petersi, X. laevis victorianus, X. (laevis) borealis, X. gilli, X. muelleri, and X. fraseri have chromosome numbers 2n=36; X. tropicalis has 20 (2n), X. (laevis) bunyoniensis 72 and X. ruwenzoriensis 108. This heterogeneity of the chromosome numbers is interesting as it represents new examples of polyploidy among Anurans. There are no big morphological differences among the karyotypes of the divers species, only the chromosomes with secondary constrictions vary considerably.