Variation in Wolbachia cidB gene,but not cidA,is associated with cytoplasmic incompatibility mod phenotype diversity in Culex pipiens

Endosymbiotic Wolbachia bacteria are, to date, considered the most widespread symbionts in arthropods and are the cornerstone of major biological control strategies. Such a high prevalence is based on the ability of Wolbachia to manipulate their hosts' reproduction. One manipulation called cytoplasmic incompatibility (CI) is based on the death of the embryos generated by crosses between infected males and uninfected females or between individuals infected with incompatible Wolbachia strains. CI can be seen as a modification‐rescue system (or mod‐resc) in which paternal Wolbachia produce mod factors, inducing embryonic defects, unless the maternal Wolbachia produce compatible resc factors. Transgenic experiments in Drosophila melanogaster and Saccharomyces cerevisiae converged towards a model where the cidB Wolbachia gene is involved in the mod function while cidA is involved in the resc function. However, as cidA expression in Drosophila males was required to observe CI, it has been proposed that cidA could be involved in both resc and mod functions. A recent correlative study in natural Culex pipiens mosquito populations has revealed an association between specific cidA and cidB variations and changes in mod phenotype, also suggesting a role for both these genes in mod diversity. Here, by studying cidA and cidB genomic repertoires of individuals from newly sampled natural C. pipiens populations harbouring wPipIV strains from North Italy, we reinforce the link between cidB variation and mod phenotype variation fostering the involvement of cidB in the mod phenotype diversity. However, no association between any cidA variants or combination of cidA variants and mod phenotype variation was observed. Taken together our results in natural C. pipiens populations do not support the involvement of cidA in mod phenotype variation.     

Mutually exclusive distribution of the sap and eag S-layer genes and the lytB/lytA cell wall hydrolase genes in Bacillus thuringiensis

Recently, two Bacillus thuringiensis strains were reported to synthesize parasporal inclusion bodies made not of the expected crystal (Cry) proteins but rather of the surface layer proteins (SLP) Sap (encoded by sap) and EA1 (encoded by eag), respectively. Whether the presence of the sap and eag genes is restricted to these two B. thuringiensis strains or ubiquitous in B. thuringiensis is unknown. We report here the distribution of the sap and eag genes in B. thuringiensis. Strains in the Bacillus cereus group were added for comparison purposes. We show that sap and eag are either present in tandem in 35% of the B. thuringiensis strains analysed and absent in 65% of the strains. When absent, a different tandem, the lytB/lytA cell wall hydrolase genes, is present. The distribution of the sap and eag S-layer and the lytB/lytA cell wall hydrolase genes is not species-specific in B. thuringiensis, B. cereus and Bacillus weihenstephanensis. Bacillus anthracis and Bacillus mycoides harbor sap and eag but not lytB/lytA. The sap, eag and lytB/lytA genes were absent in Bacillus pseudomycoides. Clearly, the distribution of the sap and eag S-layer and the lytB/lytA cell wall hydrolase genes in B. thuringiensis and in the Bacillus cereus group is mutually exclusive. We also showed that two genes involved in cell wall metabolism, csaA and csaB, are present not only upstream of the sap and eag S-layer genes, but also upstream of the lytB/lytA tandem in strains where sap and eag are absent. Bootstrapped neighbor-joining trees were inferred from the translated amino acid sequences of sap, eag and the tandem lytB/lytA, respectively.     

Comparative forelimb morphology of scratch-digging and chisel-tooth digging African mole-rat species

Bathyergus suillus (Cape dune mole-rat) and Heterocephalus glaber (naked mole-rat) are two species of subterranean burrowing rodents. Bathyergus suillus occurs in soft sandy soils and is regarded as a scratch-digger, while H. glaber is found in hard, compact soils and is a chisel-tooth digging species. The present study aimed to determine musculoskeletal differences in the forelimb of these two species. The muscles of the forelimb, back and neck were dissected to the points of origin and insertion in the left and right forelimbs, B. suillus (n = 7) and H. glaber (n = 5). Dissected muscles were photographed before maceration to demonstrate muscle attachments. The scapular spine, acromion process and clavicle were relatively straight in B. suillus. In comparison a curved scapular spine, acromion process and clavicle were observed in H. glaber. In both species, the clavicle rested on the greater tuberosity of the humerus. In B. suillus, the deltoid tuberosity was prominent and situated more distally on the humeral shaft compared to the indistinct, more proximally situated deltoid tuberosity in H. glaber. A prominent bony structure underlying the thenar pad as well as a cartilaginous protrusion beneath the hypothenar pad were observed on the palmar surface of the manus in B. suillus. Prominent claws were observed in B. suillus. A robust m. sternohyoideus was observed in H. glaber while mm. tensor fasciae antebrachii and coracobrachialis were absent. The flexors of the antebrachium of B. suillus had additional and enlarged attachment sites. The forelimb of B. suillus may be morphologically adapted for scratch-digging with relatively large and additional forelimb muscles and robust bones. In comparison, H. glaber had a reduction in the relative size, amount of muscles as well as number of attachment sites in the forelimb muscles, while the well-developed ventral neck muscles may facilitate neck and head stabilisation during chisel-tooth digging.     

Neutrophil activation by Candida glabrata but not Candida albicans promotes fungal uptake by monocytes

Candida albicans and Candida glabrata account for the majority of candidiasis cases worldwide. Although both species are in the same genus, they differ in key virulence attributes. Within this work, live cell imaging was used to examine the dynamics of neutrophil activation after confrontation with either C. albicans or C. glabrata. Analyses revealed higher phagocytosis rates of C. albicans than C. glabrata that resulted in stronger PMN (polymorphonuclear cells) activation by C. albicans. Furthermore, we observed differences in the secretion of chemokines, indicating chemotactic differences in PMN signalling towards recruitment of further immune cells upon confrontation with Candida spp. Supernatants from co‐incubations of neutrophils with C. glabrata primarily attracted monocytes and increased the phagocytosis of C. glabrata by monocytes. In contrast, PMN activation by C. albicans resulted in recruitment of more neutrophils. Two complex infection models confirmed distinct targeting of immune cell populations by the two Candida spp.: In a human whole blood infection model, C. glabrata was more effectively taken up by monocytes than C. albicans and histopathological analyses of murine model infections confirmed primarily monocytic infiltrates in C. glabrata kidney infection in contrast to PMN‐dominated infiltrates in C. albicans infection. Taken together, our data demonstrate that the human opportunistic fungi C. albicans and C. glabrata are differentially recognized by neutrophils and one outcome of this differential recognition is the preferential uptake of C. glabrata by monocytes.     

The Argasidae, Ixodidae and Nuttalliellidae (Acari: Ixodida): A World List of Valid Tick Names

The world's argasid tick fauna comprises 183 species in four genera, namely Argas, Carios, Ornithodoros and Otobius in the family Argasidae. The ixodid tick fauna consists of 241 species in the genus Ixodes and 442 species in the genera Amblyomma, Anomalohimalaya, Bothriocroton, Cosmiomma, Dermacentor, Haemaphysalis, Hyalomma, Margaropus, Nosomma, Rhipicentor and Rhipicephalus in the family Ixodidae, with the genus Boophilus becoming a subgenus of the genus Rhipicephalus. The family Nuttalliellidae is represented by the monospecific genus Nuttalliella. The species names of these ticks, based on seven previous complete or partial listings, as well as those of recently described new species, are presented in tabular format. This revised version was published online in July 2006 with corrections to the Cover Date.     

Structural organization of the nuclear ribosomal RNA genes in Cannabis and Humulus (Cannabaceae)

The structural organization of the nuclear ribosomal DNA (rDNA) of Humulus lupulus, H. japonicus and Cannabis sativa was determined by restriction site mapping. A high degree of DNA sequence similarity was evident in the coding regions of the rDNA repeats of the taxa and supports the placement of Cannabis and Humulus in one family, Cannabaceae. However, the presence of a BstEII site, an additional SacI site, absence of the SpeI site and positional differences of the SspI sites in the 25 S gene distinguished H. japonicus from H. lupulus. Humulus lupulus has an additional EcoRV site in the IGS region. A XhoI site in the 18S region of C. sativa distinguishes it from the two hop species. The diagnostic differences in the IGS of C. sativa include the EcoRI, HindIII and XhoI sites. These sites were not detected in the IGS of the two hop species.     

Inflorescence development in the Vitis–Ampelocissus clade of Vitaceae: the unusual lamellate inflorescence of Pterisanthes

Pterisanthes (Vitaceae) is a genus of c. 20 species of scandent climbers endemic to Southeast Asia with unusual lamellate inflorescences. Molecular phylogenetic analysis supports its relationship in the well‐supported Vitis–Ampelocissus–Nothocissus–Pterisanthes clade (i.e. the Ampelocissus–Vitis clade). Shoot tips and floral buds were collected from wild and greenhouse‐grown P. eriopoda at different developmental stages and were examined using epi‐illumination, light and scanning electron microscopy. Inflorescence and floral ontogeny was studied to discover how the lamellate inflorescence evolved and to make morphological comparisons to infer relationships with closely related members of Vitaceae. The second‐order branches in P. eriopoda are racemose and develop helically around the inflorescence axis in a similar fashion to Vitis and Ampelocissus. Inflorescence branching is restricted to the second order in P. eriopoda, whereas in Vitis and most Ampelocissus species subsequent branching orders culminate in the typical vitaceous determinate dichasium. In P. eriopoda subsequent lateral growth of the second‐order branches combined with the inhibition of peduncle or pedicel formation and loss of dichasial branching results in the unique lamellae in Pterisanthes, on which the floral primordia arise directly in a helical pattern. Floral development in P. eriopoda is the same as in other genera of Vitaceae examined to date with initiation of floral whorls centripetally, the calyx ring developing first and calyx lobes fused to cover the petals and stamen primordia. Given the recent phylogenetic results that placed Pterisanthes firmly within Ampelocissus, the most likely scenario is that the Pterisanthes inflorescence is derived from the thyrse of an Ampelocissus‐like ancestor and that the thyrse is a morphological synapomorphy of the Ampelocissus–Vitis clade. © 2015 The Linnean Society of London, Botanical Journal of the Linnean Society, 2015, 179 , 725–741.     

Crinkled employs wingless pathway for wing development in Tribolium castaneum

Crinkled is associated with embryonic denticle formation and auditory organ development in Drosophila melanogaster. However, the functions of Crinkled have not been fully investigated. Additionally, the genes that participate in the Crinkled pathway are unknown. Phylogenetic analysis indicates that crinkled exhibits a one‐to‐one orthologous relationship in insects. In Tribolium castaneum, the crinkled gene is 6,498 bp in length and consists of six exons. Crinkled expression peaked during two phases in Tribolium: late embryonic and pupal stages. High levels of crinkled mRNA were detected in the fat body, head, epidermis, ovary, and accessory gland of late adults. Knockdown of crinkled using RNA interference (RNAi) severely affected wing morphogenesis in T. castaneum. We further showed that crinkled silencing reduced forked expression through wingless and shaven‐baby, and RNAi of forked phenocopied the effects of crinkled knockdown in T. castaneum. This study investigated the development role of crinkled in postembryonic stages and indicated that forked mediates the functions of crinkled during wing morphogenesis in T. castaneum.     

Opalinidae in African Anura. II. Genera Protozelleriella n. g. and Zelleriella

During 1988 and 1989, 409 specimens of southern African Anura, comprising 50 species in 9 families, were checked for opalinids in the cloaca. Protozelleriella devilliersi n. g., n. sp. was found in three of four Capensibufo rosei (Bufonidae); Zelleriella africana Sandon, 1938 in two of three Bufo angusticeps and one of 20 B. garmani (Bufonidae) plus three of 12 Phrynobatrachus natalensis, 12 of 19 Tomopterna cryptotis, seven of 14 T. delalandii, three of seven T. krugerensis and one of six T. natalensis (all Ranidae); Z. gambieri n. sp. in one of six T. natalensis; Z. garlandi n. sp. in two of six Kassina maculata (Hyperoliidae); Z. sandoni n. sp. in two of 20 B. garmani, two of six K. maculata, one of seven K. senegalensis and one of four Leptopelis mossambicus (Hyperoliidae) plus one of five Phrynomerus bifasciatus (Microhylidae) and two of 19 T. cryptotis. It is suggested that the Ranidae (in particular the genus Tomopterna) and the Hyperoliidae are among the major carriers of Zelleriella in the Afrotropical Region.     

河滨湿地不同植物群落根系分布特征与土壤理化性状的关系——以黄河中游荥阳段为例

在黄河中游郑州荥阳段,选择了5种河滨湿地植物群落进行根系和土壤性状特征研究,以期阐明不同植物群落的根系分布规律与土壤性状的关系,为河滨湿地植物群落组成以及土壤质量恢复提供科学参考。结果表明(1)在0—40 cm土层,根生物量密度与根长密度的平均值均表现为：芦苇群落(Phragmites australis)和芦苇-狗牙根(Cynodon dactylon)群落均大于芦苇-拂子茅(Calamagrostis epigeios)-狗牙根群落、拂子茅-狗牙根群落、拂子茅-狗牙根-水莎草(Juncellus serotinus)群落。拂子茅-狗牙根、芦苇-拂子茅-狗牙根、拂子茅-水莎草-狗牙根三种植物群落类型下根生物量密度、根长密度在0—20 cm表层土壤较大,芦苇群落和芦苇-狗牙根群落的根生物量密度、根长密度在10—40 cm的土层较大。(2)黄河河滨湿地芦苇群落、芦苇-狗牙根群落的土壤以粉粒为主,拂子茅-狗牙根群落、芦苇-拂子茅-狗牙根群落、拂子茅-狗牙根-水莎草群落的土壤主要以砂粒为主。在0—40 cm土层,芦苇群落、芦苇-狗牙根群落的土壤含水率、土壤有机质、有效氮和有效磷含量均显著高于...     

Grosmannia and Leptographium spp. associated with conifer-infesting bark beetles in Finland and Russia, including Leptographium taigense sp. nov.

Species of Grosmannia with Leptographium anamorphs include important forest pathogens and agents of blue stain in timber. They are commonly found in association with forest pests, such as bark beetles. During a survey of ophiostomatoid fungi in eastern parts of Finland and neighboring Russia, species belonging to the genus Grosmannia were isolated from 12 different bark beetle species infesting Picea abies and Pinus sylvestris, the most economically important conifers in the region. Identification of these fungi was based on morphology, DNA sequence comparisons for three gene regions and phylogenetic analyses. A total of ten taxa were identified. These belonged to six different species complexes in Grosmannia. The phylogenetic analyses provided an opportunity to redefine the G. galeiformis-, L. procerum-, L. lundbergii-, G. piceiperda-, G. olivacea- and G. penicillata-complexes, and to consider the species emerging from the survey within the context of these complexes. The species included G. galeiformis, G. olivacea, L. chlamydatum, L. lundbergii, L. truncatum and a novel taxon, described here as L. taigense sp. nov. In addition, species closely related to G. cucullata, G. olivaceapini comb. nov., G. piceiperda and L. procerum were isolated but their identity could not be resolved. The overall results indicate that the diversity of Grosmannia species in the boreal forests remains poorly understood and that further studies are needed to clarify the status of several species or species complexes.     

Morphological features and chromosome numbers in cultures of five Cephaleuros species (Trentepohliaceae,Chlorophyta) from Japan

The morphological features and chromosome numbers were examined in cultures of five species of Cephaleuros; C. aucubae, C. biolophus, C. japonicus, C. microcellularis, and C. virescens, collected from Japan. On agar media of Bold's basal medium and CA medium, radial growth of the algal colonies was vigorous in C. virescens, moderate in C. aucubae, and poor in C. microcellularis, but varied from poor to moderate in isolates of C. biolophus and C. japonicus. Filaments of C. virescens branched at an angle less than 40° and just below the cross walls, while those of the other species branched at an angle greater than 40° without relation to the cross walls. The length of filamentous cells was significantly smaller in C. microcellularis than in the other species. The chromosome number differed with the species: n = 22 in C. aucubae, n = 34 in C. biolophus, n = 18 in C. japonicus, n = 12 in C. microcellularis, and n = 24 in C. virescens. The chromosome number did not vary between isolates from filaments and gametes in C. aucubae and C. biolophus and among those from filaments, gametes, and zoospores in C. japonicus and C. virescens.     

Genetic control of leaf-blade morphogenesis by the <Emphasis Type="Italic">INSECATUS</Emphasis> gene in <Emphasis Type="Italic">Pisum sativum</Emphasis>

To understand the role of INSECATUS (INS) gene in pea, the leaf blades of wild-type, ins mutant and seven other genotypes, constructed by recombining ins with uni-tac, af, tl and mfp gene mutations, were quantitatively compared. The ins was inherited as a recessive mutant allele and expressed its phenotype in proximal leaflets of full size leaf blades. In ins leaflets, the midvein development was arrested in distal domain and a cleft was formed in lamina above this point. There was change in the identity of ins leaflets such that the intercalary interrupted midvein bore a leaf blade. Such adventitious blades in ins, ins tl and ins tl mfp were like the distal segment of respective main leaf blade. The ins phenotype was not seen in ins af and ins af uni-tac genotypes. There was epistasis of uni-tac over ins. The ins, tl and mfp mutations interacted synergistically to produce highly pronounced ins phenotype in the ins tl mfp triple mutant. The role(s) of INS in leaf-blade organogenesis are: positive regulation of vascular patterning in leaflets, repression of UNI activity in leaflet primordia for ectopic growth and in leaf-blade primordium for indeterminate growth of rachis, delimitation of proximal leaflet domain and together with TL and MFP homeostasis for meristematic activity in leaflet primordia. The variant apically bifid shape of the affected ins leaflets demonstrated that the leaflet shape is dependent on the venation pattern.     

Variation analysis of pathogenic Vibrio spp. and Pseudomonas spp. in Changhai mollusc farming waters using real-time PCR assay during 2011–2014

Bacterial disease has caused high mortality of breeding molluscs from 2009 to 2011 in the Changhai area (Dalian, China). Vibrio spp. and Pseudomonas spp. have been detected as major pathogenic agents for aquatic animals in this area. In the present study, four virulence genes including vsm, toxR, aprX and carA were targeted to develop a real-time PCR assay for the quantitative detection of Vibrio splendidus, V. parahaemolyticus, Pseudomonas fluorescens and P. putida, respectively. The sensitivity and specificity of the assays were verified by experimental samples, and the variation tendencies of pathogenic V. splendidus, V. parahaemolyticus, P. fluorescens and P. putida strains from June to September were also detected in mollusc farming waters during 2011–2014. The concentration of V. splendidus increased from June to July, reduced in August, and then increased again in September. The highest count of V. parahaemolyticus appeared in July, and then dramatically decreased from August to September. Conversely, the counts of P. fluorescens and P. putida remained at lower levels from June to August, and then dramatically peaked in September. All four pathogenic bacteria displayed similar fluctuation tendencies of count variation in each year, and their concentrations were found to have a correlation with the average annual temperature. The variation tendency of pathogenic bacteria with temperature suggested that temperature was one of the most important factors to regulate the bacterial growth in a farming area, which could further provide information for the early warning of disease outbreak by using a convenient real-time PCR assay.     

Vriesea friburgensis: A natural trap or a nurse plant in coastal sand dunes?

Positive interactions between plants play a fundamental role in environments with extreme conditions, such as coastal sand dunes, where the establishment of many species is limited by high temperature, and low availability of water and nutrients in the soil. The ability to store water and enrich the soil with nutrients characterizes bromeliads as potential nurse plants for other species. This study aimed to analyze the potential of the bromeliad Vriesea friburgensis to act as a nurse plant, and to compare this interaction in coastal dunes in herbaceous versus scrub vegetation. The study was performed on Santa Catarina Island in Brazil. Juveniles of other species associated with V. friburgensis were collected within and under bromeliad tanks and on adjacent plots beside each bromeliad. The abundance, species richness and height of juveniles were recorded. We found that abundance and richness of Vf‐associated juveniles were lower than those Vf‐not associated, in herbaceous vegetation (negative interaction). However, the height of some shrubs species, like Eupatorium casarettoi and Tibouchina urvilleana, was greater when Vf‐associated (positive interaction). In scrub vegetation, the abundance and richness of juveniles were not significantly affected by V. friburgensis presence (neutral interaction). However, the height of some shrubs and trees species, like Clusia criuva, Eugenia catharinae, Myrsine parvifolia and Ocotea pulchella, was greater when Vf‐associated. Species composition associated with V. friburgensis differed from the assemblage in adjacent plots, indicating that some species interact more with nurse plants, whereas others fail to develop when associated with this bromeliad; the difference between groups in species composition was less evident in scrub vegetation areas. Clusia criuva was considered an indicator species for Vf‐associated juveniles in both vegetation types. Despite the fact that these tank bromeliads do not function as nurse plants for some species, there are others that seem to be favoured by this interaction.     

Phylogenetic relationships in Carex, subgenus Vignea (Cyperaceae), based on ITS sequences

To evaluate the sectional classification in Carex, subgenus Vignea, the ITS region of 58 species of 20 sections was analyzed with Neighbor Joining (NJ) and Markov chain Monte Carlo (MCMC) methods. Sections Dioicae, Physodeae and Ovales are found to be monophyletic, with C. bohemica well integrated in the section Ovales. Section Heleonastes turns out to be monophyletic, if C. canescens is treated separately in section Canescentes. Section Elongatae is monophyletic, but C. remota is placed in section Remotae and C. bromoides in section Deweyanae. In both analyses, six representatives of section Arenariae cluster together in a terminal group, whereas C. disticha, C. repens and C. siccata form a basal cluster. C. maritima, as the only member of section Incurvae, shares this basal position. C. chordorrhiza is ascribed to section Chordorrhizeae and not ascribed to the paraphyletic section Divisae. C. vulpina and C. otrubae are assigned to section Vulpinae and separated from the heterogeneous section Stenorhynchae. The other members of sections Divisae, Muehlenbergianae, Multiflorae and Stenorhynchae are scattered throughout the trees. The representatives of section Foetidae are dispersed in both analyses, section Paniculatae appears to be non-monophyletic in the molecular results as well. The subgenus appears subdivided in at least four larger subgroups in all analyses. Whereas these subgroups are strongly supported, the relationships between these subgroups remain only poorly resolved.     

Phylogenetic relationships of the chloroplast genomes in the genus Glycine inferred from four intergenic spacer sequences

The nucleotide sequences of four intergenic spacer regions of chloroplast DNA, atpB-rbcL, trnS-trnG, rps11-rpl36, and rps3-rpl16, were analyzed in the genus Glycine. Phylogenetic analysis based on the sequence data using Neonotonia wightii as the outgroup generated trees supporting the classification of two subgenera, Soja and Glycine, and three plastome groups in the subgenus Glycine. The results were consistent with the presence of diversified chloroplast genomes within tetraploid plants of G. tabacina and G. tomentella, as well as with a close relationship between G. tomentella and G. dolichocarpa that had been suggested based on morphological analyses. Little sequence variation was found in the subgenus Soja, suggesting that G. soja rapidly expanded its distribution in East Asia. The analysis also showed that the differentiation into three plastome groups in the subgenus Glycine occurred in the early stages of its evolution, after the two subgenera diverged.     

Phylogenetic Analysis of S-Locus Genes Reveals the Complicated Evolution Relationship of S Haplotypes in Brassica

Self-incompatibility (SI) is reported to play a key role in the evolution of species as it promotes their outcrossing through the recognition and rejection of self-pollen grains. In Brassica, two S-locus genes expressed in the stigma, S-locus glycoprotein (SLG) gene and S-locus receptor kinase (SRK) gene, and one expressed in the pollen, S-locus protein 11 (SP11) gene, were linked as an S haplotype. In order to analyze the evolutionary relationships of S haplotypes in Brassica, a total of 39 SRK, 37 SLG, and 58 SP11 sequences of Brassica oleracea, Brassica rapa and Brassica napus were aligned. Two phylogenetic trees with similar pattern were constructed based on the nucleotide sequences of SRK/SLG and SP11, respectively. Class I and class II alleles were clustered into two distinct groups, and alleles from different species, including all the interspecific pairs of S haplotypes, were closely related to each other. The S-locus genes identified in B. napus were intermingled in phylogenetic trees. All these observations showed that class I and class II S haplotypes diverged ahead of the species differentiation in Brassica. The evolution and the genetic diversity of S haplotypes in Brassica were discussed. Moreover, the relationships between S haplotypes and SI phenotypes in Brassica, especially in B. napus, were also discussed.     

Opalinidae in African Anura. III. Genus Cepedea

During 1988 and 1989, 409 specimens of southern African Anura comprising 50 species in 9 families were checked for opalinids in the cloaca. Cepedea acuta n. sp. was found in six of 19 Tomopterna cryptotis and four of seven T. krugerensis (Ranidae); C. affinis (Nazaretskaja, 1992) in two of four Afrixalus aureus, two of four Hyperolius horstocki, 23 of 42 H. marmoratus, one of seven H. pusillus, two of six H. semidiscus, and eight of 12 H. tuberilinguis (Hyperoliidae); C. magna Metcalf, 1923 in one of 20 Bufo garmani, eight of 33 B. gutturalis and two of nine B. rangeri (all Bufonidae), one of two Heleophryne natalensis (juveniles) (Heleophrynidae), four of six Kassina maculata, three of seven K. senegalensis and two of six Semnodactylus wealii (all Hyperoliidae), three of five Phrynomerus bifasciatus (Microhylidae), three of 12 Phrynobatrachus natalensis, 11 of 19 Tomopterna cryptotis, 13 of 14 T. delalandii, one of seven T. krugerensis and one of six T. natalensis (all Ranidae), and four of five Chiromantis xerampelina (Rhacophoridae); Cepedea vanniekerkae n. sp. in one of 19 Tomopterna cryptotis. It is suggested that the Ranidae (in particular the genus Tomopterna) and the Hyperoliidae (in particular the genus Hyperolius) are among the major carriers of Cepedea in the Afrotropical Region.