Michael F. Barej

Take time to smell the frogs: vocal sac glands of reed frogs (Anura: Hyperoliidae) contain species-specific chemical cocktails

Males of all reed frog species (Anura: Hyperoliidae) have a prominent, often colourful, gular patch on their vocal sac, which is particularly conspicuous once the vocal sac is inflated. Although the presence, shape, and form of the gular patch are well-known diagnostic characters for these frogs, its function remains unknown. By integrating biochemical and histological methods, we found strong evidence that the gular patch is a gland producing volatile compounds, which might be emitted while calling. Volatile compounds were confirmed by gas chromatography–mass spectrometry in the gular glands in 11 species of the hyperoliid genera Afrixalus, Heterixalus, Hyperolius, and Phlyctimantis. Comparing the gular gland contents of 17 specimens of four sympatric Hyperolius species yielded a large variety of 65 compounds in species-specific combinations. We suggest that reed frogs might use a complex combination of at least acoustic and chemical signals in species recognition and mate choice.

Batrachochytrium dendrobatidis in amphibians of Cameroon, including first records for caecilians

Amphibian chytrid fungus Batrachochytrium dendrobatidis (Bd) has been hypothesised to be an indigenous parasite of African amphibians. In Cameroon, however, previous surveys in one region (in the northwest) failed to detect this pathogen, despite the earliest African Bd having been recorded from a frog in eastern Cameroon, plus one recent record in the far southeast. To reconcile these contrasting results, we present survey data from 12 localities across 6 regions of Cameroon from anurans (n = 1052) and caecilians (n = 85) of ca. 108 species. Bd was detected in 124 amphibian hosts at 7 localities, including Mt. Oku, Mt. Cameroon, Mt. Manengouba and lowland localities in the centre and west of the country. None of the hosts were observed dead or dying. Infected amphibian hosts were not detected in other localities in the south and eastern rainforest belt. Infection occurred in both anurans and caecilians, making this the first reported case of infection in the latter order (Gymnophiona) of amphibians. There was no significant difference between prevalence and infection intensity in frogs and caecilians. We highlight the importance of taking into account the inhibition of diagnostic qPCR in studies on Bd, based on all Bd-positive hosts being undetected when screened without bovine serum albumin in the qPCR mix. The status of Bd as an indigenous, cosmopolitan amphibian parasite in Africa, including Cameroon, is supported by this work. Isolating and sequencing strains of Bd from Cameroon should now be a priority. Longitudinal host population monitoring will be required to determine the effects, if any, of the infection on amphibians in Cameroon.

No ecological opportunity signal on a continental scale? Diversification and life-history evolution of African true toads (Anura: Bufonidae).

The niche‐filling process predicted by the “ecological opportunity” (EO) model is an often‐invoked mechanism for generating exceptional diversity in island colonizers. Whether the same process governs lineage accumulation and trait disparity during continental colonization events is less clear. Here, we test this prediction by investigating the rate dynamics and trait evolution of one of Africas most widespread amphibian colonizers, the true toads (Bufonidae). By reconstructing the most complete molecular phylogeny of African Bufonidae to date, we find that the diversification of lineages in Africa best conforms to a constant rate model throughout time and across subclades, with little support for EO. Evolutionary rates of life‐history traits have similarly been constant over time. However, an analysis of generalists and specialists showed a shift toward higher speciation rates associated with habitat specialization. The overall lack of EO signal can be interpreted in a number of ways and we propose several explanations. Firstly, methodological issues might preclude the detection of EO. Secondly, colonizers might not experience true EO conditions and due to the size, ecological heterogeneity and age of landmasses, the diversification processes might be more complex. Thirdly, lower speciation rates of habitat generalists may have affected overall proliferation of lineages.

Light shines through the spindrift - Phylogeny of African torrent frogs (Amphibia, Anura, Petropedetidae)

Torrent frogs of the genus Petropedetes Reichenow, 1874 as currently understood have a disjunct distribution with species endemic to West, Central or East Africa. We herein present a phylogenetic analysis including all but one of the currently described 12 species of the genus. Maximum Likelihood and Bayesian analyses of combined nuclear (rag1, SIA, BDNF) and mitochondrial (16S, 12S, cytb) genes of more than 3500 base pairs, revealed clades corresponding to the three sub-Saharan regions. Molecular results are confirmed by morphological differences. Surprisingly, the three geographic clades do not form a monophyletic group with respect to closely related families Pyxicephalidae and Conrauidae and therefore require taxonomic changes. We resurrect Arthroleptides Nieden, 1911 for the East African taxa. The Central African taxa remain in the genus Petropedetes. The West African members are placed in the new genus Odontobatrachus gen. nov. The taxonomic position of the new genus remains incertae sedis as it was not assigned to any of the four families included in our analyses. Potential new species have been detected within all three major clades, pointing to a still not fully clarified diversity within African torrent frogs.

Evaluating mechanisms of diversification in a Guineo-Congolian tropical forest frog using demographic model selection

The accumulation of biodiversity in tropical forests can occur through multiple allopatric and parapatric models of diversification, including forest refugia, riverine barriers and ecological gradients. Considerable debate surrounds the major diversification process, particularly in the West African Lower Guinea forests, which contain a complex geographic arrangement of topographic features and historical refugia. We used genomic data to investigate alternative mechanisms of diversification in the Gaboon forest frog, Scotobleps gabonicus, by first identifying population structure and then performing demographic model selection and spatially explicit analyses. We found that a majority of population divergences are best explained by allopatric models consistent with the forest refugia hypothesis and involve divergence in isolation with subsequent expansion and gene flow. These population divergences occurred simultaneously and conform to predictions based on climatically stable regions inferred through ecological niche modelling. Although forest refugia played a prominent role in the intraspecific diversification of S. gabonicus, we also find evidence for potential interactions between landscape features and historical refugia, including major rivers and elevational barriers such as the Cameroonian Volcanic Line. We outline the advantages of using genomewide variation in a model‐testing framework to distinguish between alternative allopatric hypotheses, and the pitfalls of limited geographic and molecular sampling. Although phylogeographic patterns are often species‐specific and related to life‐history traits, additional comparative studies incorporating genomic data are necessary for separating shared historical processes from idiosyncratic responses to environmental, climatic and geological influences on diversification.

Idiosyncratic responses to climate‐driven forest fragmentation and marine incursions in reed frogs from Central Africa and the Gulf of Guinea Islands

Organismal traits interact with environmental variation to mediate how species respond to shared landscapes. Thus, differences in traits related to dispersal ability or physiological tolerance may result in phylogeographic discordance among co‐distributed taxa, even when they are responding to common barriers. We quantified climatic suitability and stability, and phylogeographic divergence within three reed frog species complexes across the Guineo‐Congolian forests and Gulf of Guinea archipelago of Central Africa to investigate how they responded to a shared climatic and geological history. Our species‐specific estimates of climatic suitability through time are consistent with temporal and spatial heterogeneity in diversification among the species complexes, indicating that differences in ecological breadth may partly explain these idiosyncratic patterns. Likewise, we demonstrated that fluctuating sea levels periodically exposed a land bridge connecting Bioko Island with the mainland Guineo‐Congolian forest and that habitats across the exposed land bridge likely enabled dispersal in some species, but not in others. We did not find evidence that rivers are biogeographic barriers across any of the species complexes. Despite marked differences in the geographic extent of stable climates and temporal estimates of divergence among the species complexes, we recovered a shared pattern of intermittent climatic suitability with recent population connectivity and demographic expansion across the Congo Basin. This pattern supports the hypothesis that genetic exchange across the Congo Basin during humid periods, followed by vicariance during arid periods, has shaped regional diversity. Finally, we identified many distinct lineages among our focal taxa, some of which may reflect incipient or unrecognized species.

A New Small Phrynobatrachus (Amphibia: Anura: Phrynobatrachidae) From Southern Cameroon

We describe a new small Phrynobatrachus species from southern Cameroon. The new species exhibits a combination of unique morphological characters and a distinctive colour pattern consisting of a black lateral face mask, a black throat in males, a white throat with uniform black lower mandibles in females and a white belly in both sexes. Morphologically it is characterized by small size, absence of an eyelid cornicle, presence of black spines on anterior part of vocal sac in males, spines on flanks, presence of discs on toe and finger tips, distinct webbing, absence of nuptial pads on male thumbs and scapular ridges converging in a straight line. Analysis of mitochondrial 16S rRNA revealed that the new species differs from 34 other West and Central African species of the genus by a minimum distance of 4.5% and is most similar to several Phrynobatrachus species which are almost all endemic to the Cameroon volcanic line or parts of it, i.e. P. chukuchuku (4.9%), P. werneri (5.1%), P. steindachneri (5.2%), P. schioetzi (5.6%), P. batesii (5.9%), P. cricogaster (5.5%), P. danko (6.1%), and P. manengoubensis (6.1%), respectively. The new species is most similar to P. batesii, which was described from forests close to the type locality of the new species. Morphologically the new species differs from P. batesii by much smaller size (< 20 mm vs. 25-31 mm), shape of scapular ridges, belly colour, shorter shanks, absence of nuptial pads and presence of gular spines in breeding males.

Two new Phrynobatrachus species (Amphibia: Anura: Phrynobatrachidae) from the Republic of the Congo

We describe two new species of puddle frogs, genus Phrynobatrachus, from the south-western Republic of the Congo. One of them, P. horsti sp. nov., occurs also in neighbouring Gabon and is morphologically most similar to the Cameroonian P. ruthbeateae. It differs from the latter species by smaller males with longer thighs and shanks. The new species comprises various colour morphs but always has less conspicuous black borders between flanks and belly than P. ruthbeateae. The distinct and large black axillary blotch of P. ruthbeateae is either much smaller in P. horsti sp. nov., or broken into numerous irregularly shaped smaller dots. Similarly, a black transversal line at the anterior ventral border of thighs and the black face mask is less distinct and irregularly delimitated in P. horsti sp. nov. when compared to P. ruthbeateae. The mean genetic difference in the sampled region of the 16S rRNA gene between P. horsti sp. nov. and 40 other western African congeners range from 3.66-18.10%. The second new species, P. mayokoensis sp. nov., differs from all other known congeners by the combination of a compact and warty body, the absence of a spiny eyelid tubercle and pedal webbing, a conspicuous black triangle on throat and anterior part of the belly, and a distinct large red blotch on the anterior-proximal surface of the thighs. It exhibited a mean genetic difference in the 16S rRNA to 40 other western African congeners ranging from 1.34-16.98%. The genetically most similar sequence stems from a GenBank entry of a Gabonese frog, determined as P. ogoensis. A comparison of the new species with P. ogoensis syntypes confirmed their specific distinctiveness, most convincingly underlined by the absence of pedal webbing in the new species and the pronounced pedal webbing in P. ogoensis. The GenBank entry thus most likely is based on a misidentification and P. mayokoensis sp. nov. may also occur in neighbouring Gabon. The discovery of the two new frog species is further evidence of the huge gap in our knowledge concerning the species richness in the Guineo-Congolian rainforests.

The tadpoles of nine Cameroonian Leptodactylodon species (Amphibia, Anura, Arthroleptidae)

We describe and compare the tadpoles of nine Leptodactylodon species from Cameroon. The tadpoles of Leptodactylodon bicolor, L. mertensi, L. ovatus, L. perreti and L. ventrimarmoratus are herein reinvestigated, partly based on larger series than previously available. In addition we present first descriptions for the tadpoles of L. boulengeri, L. erythrogaster, L. ornatus, and L. cf. polyacanthus. The morphology of these exotrophic, lotic and neustonic tadpoles is discussed in comparison with other stream-dwelling tadpoles. Based on the assumed biology of these tadpoles, living in interstices of gravel or debris, the functioning of several special morphological features, in particular the funnel-mouth of Leptodactylodon tadpoles, are interpreted.

Relicts of a forested past: Southernmost distribution of the hairy frog genus Trichobatrachus Boulenger, 1900 (Anura: Arthroleptidae) in the Serra do Pingano region of Angola with comments on its taxonomic status

The monotypic genus Trichobatrachus, with its sole representative, the hairy frog, Trichobatrachus robustus Boulenger, 1900, could be considered one of the most well-known frogs of Africa. Despite its broad recognition and the fact that it is considered wide-spread and locally common (Amiet & Burger 2004), surprisingly little is known about the actual distribution and the specific occurrence patterns of the species. It was originally described by Boulenger (1900) from the Benito River, previously erroneously stated to be located in Gabon. However, Lötters et al. (2001) clarified that the type locality was actually situated in Equatorial Guinea, then part of French Congo, and subsequently provided the first confirmed country record for Gabon. Hairy frogs were previously included in the herpetofaunal lists of Cameroon (Parker 1936; Perret & Mertens 1957), the Democratic Republic of Congo (Laurent 1956), and Nigeria (Schiøtz 1963). More recently published accounts list them for Cameroon (Euskirchen et al. 1999; Herrmann et al. 2005; Gonwouo & Rödel 2008), Equatorial Guinea (De la Riva 1994; Lasso et al. 2002), Gabon (Lötters et al. 2001; Rödel & Pauwels 2003; Burger et al. 2004; Pauwels & Rödel 2007; Bell et al. 2011), and the Democratic Republic of Congo (DRC, Fretey et al. 2011), and speculate about their potential occurrence in Congo and the Cabinda enclave of Angola (Amiet & Burger 2004). The DRC record so far represented the most southerly distribution, while the most northerly account comes from the Adamaoua Province of northern Cameroon (Tadpole voucher MHNG-AMP/ERPI-1035.006 from Bénoué source, Northern cliff Ngaoundéré, Adamaoua, Cameroon/Adamaoua Province, cf. Perret 1966). Specimens stored in the collection at the Royal Museum of Central Africa, Tervuren (RMCA) under collection numbers RMCA B 90060.0004-11 confirm the occurrence of the species in Congo. To our knowledge, these specimens represent yet unpublished first country records.