Intergrading between members of the "regularis group" of toads in South Africa

The mating calls of three members of the " regularis group" of toads (Anura: Bufonidae) that are sympatric in part of South Africa are examined. A naturally occurring population of hybrid toads from Pretoria (South Africa) has previously been described as representing an intergrading between Bufo garmani Meek and Bufo rangeri Hewitt. Examination of the mating calls of these toads provides little evidence of intergrading between Bufo rangeri and Bufo garmani , and indicates the involvement of Bufo regularis Reuss in the hybridization. The mating calls of the hybrids are varied, and are structurally intermediate between those of the parental species. Mating call parameters are found to be useful in separating forms that are difficult to distinguish on morphological grounds alone. They are further found to be a sensitive non-morphological means of ascertaining the affinities of naturally occurring hybrid toads.     

Repetitive DNA and Feulgen hydrolysis kinetics in three species of Bufo.

Two North American species of the genus Bufo (Bufo cognatus and Bufo boreas, 2n = 22) and one African species (Bufo regularis, 2n = 20) were analyzed with respect to their repetitive DNA fractions and the behaviour of their chromatin to the acid hydrolysis at different times. The mean melting point of the total isolated DNA decreased from 89 degrees C to 87 degrees C with a genome size increase from 4.4 to 7.5 pg. The differences in genome size can only partly be explained on the basis of repetitive DNA fractions (renaturing up to Cot 10 in 0.12 M phosphate buffer). Several fractions in this repetitive range behave independently in the three species and the spectrum of repetitive fractions in the African Bufo regularis differs distinctly from those of the American toads. When fixed chromatin of these species in histochemical preparations is hydrolyzed with 5N HCl during the Feulgen reaction, the kinetics of depurination are equal in all species, while hydrolytic DNA breakdown proceeds distinctly more slowly in Bufo reularis as compared to the other species.     

Historical biogeography of amphibian parasites, genus Polystoma (Monogenea: Polystomatidae)

Aim  The present-day geographical distribution of parasites with a direct biological life cycle is guided mostly by the past dispersal and vicariance events that have affected their hosts. The Amphibia– Polystoma association (which satisfies these criteria) also exhibits original traits, such as host specificity and world-wide distribution. This biological model was thus chosen to investigate the common historical biogeography of its widespread representatives.
Location  North and South America, Eurasia and Africa.
Methods  We investigated the phylogeny of 12 species of neobatrachian parasites sampled from North and South America, Eurasia and Africa. Hosts belonged mostly to hyloids and ranoids of families Bufonidae, Hylidae, Leptodactylidae, Ranidae and Hyperoliidae. Phylogenetic reconstructions were inferred from maximum likelihood and maximum parsimony analyses from complete ITS1 sequences.
Results  The group of American species appeared paraphyletic with one species at the base of a Eurafrican clade, within which two lineages were seen: one composed of only Eurasian species, and the other of European and African species, with the two European species basal to an African clade.
Main conclusions  The route of Polystoma evolution is deduced from the phylogenetic tree and discussed in the light of host evolution. We conclude that Polystoma originated in South America on hyloids, after the separation of South America from Africa. The genus must have colonized North America in Palaeocene times and Eurasia by the mid-Cainozoic, taking advantage of the dispersal of either ancestral bufonids or hylids. Africa, however, appears to have been colonized more recently, during the Messinian period.     

Effects of season and temperature acclimation on electrocardiogram and heart rate of toads

Electrocardiogram (ECG) was recorded from unrestrained toads of two species, one resistant (Bufo viridis) and the other sensitive (B. regularis) to low temperatures. Although the temperature-sensitive species could not survive at temperatures below 7-8 degrees C, heart rate is linearly and similarly related to ambient temperature in the two species. In B. viridis, the cold resistant species, heart rate in winter was 25% lower than in summer, and the dependency of heart rate on temperature was reduced by 50% in toads acclimated to 10 degrees C in winter. It was not possible to disclose any effect of acclimation on heart rate in summer, due to the large variation in the recorded values. The QRS amplitude in the ECG was considerably reduced at low temperature only in B. viridis. It is concluded that differences in cardiac activity cannot account for the distinct difference in thermal relations of the two species, and that it should reside at other regulatory levels.     

Inhibition By Vitamin A of Cyst Formation In Nyctotheroides Puytoraci Induced By Injecting Its Host Bufo Regularis With 20-Methylcholanthrene

SYNOPSIS. 20-Methylcholanthrene induced cyst formation in Nyctotheroides puytoraci when injected into its host Bufo regularis. Presumably this hydrocarbon or its metabolites reaches the parasites in the recta of treated host animals and induces encystment. However, injection of B. regularis with 0.5 mg of 20-methylcholanthrene + vitamin A palmitate (5,000 IU) inhibited the hydrocarbon-induced encystment of the parasites.     

Origin and evolution of African Polystoma (Monogenea: Polystomatidae) assessed by molecular methods

Among Polystomatidae (Monogenea), the genus Polystoma, which mainly infests neobatrachian hosts, is the most diverse and occurs principally in Africa, from where half the species have been reported. Previous molecular phylogenetic studies have shown that this genus originated in South America, and later colonised Eurasia and Africa. No mention was made on dispersal corridors between Europe and Africa or of the origin of the African Polystoma radiation. Therefore, a molecular phylogeny was inferred from ITS1 sequences of 21 taxa comprising two species from America, seven representatives from Europe and 12 from Africa. The topology of the phylogenetic tree reveals that a single event of colonisation took place from Europe to Africa and that the putative host carrying along the ancestral polystome is to be found among ancestral pelobatids. Percentage divergences estimates suggest that some presumably distinct vesicular species in unrelated South African anurans and some neotenic forms found in several distinct hosts in Ivory Coast, could, in fact, belong to two single polystome species parasitising divergent hosts. Two main factors are identified that may explain the diversity of African polystomes: (i), we propose that following some degree of generalism, at least during the juvenile stages of both hosts and parasites, distinctive larval behaviour of polystomes engenders isolation between parasite populations that precludes sympatric speciations; (ii), cospeciation events between Ptychadena hosts and their parasites are another factor of diversification of Polystoma on the African continent. Finally, we discuss the systematic status of the Madagascan parasite Metapolystoma, as well as the colonisation of Madagascar by the host Ptychadena mascareniensis.     

Effect of ethylenediaminetetraacetic acid (EDTA) on acute mercury poisoning of toad

Studies have been carried out on the ability of EDTA (CaNa2-ethylenediaminetetraacetate) to offset the acute toxicity of mercuric chloride. The lethality of mercury was tested in female Bufo regularis species (Amphibia) which were given EDTA immediately after the Hg2+ injection. The results show that the inherent toxicity of the EDTA-mercury complex is sufficiently high for it to be ineffective as an antidote.     

A new species of Polystoma (Monogenea: Polystomatidae) parasitic in Hyperolius marmoratus (Anura: Hyperoliidae) in South Africa

Polystoma marmorati n. sp. is described as a new polystomatid species parasitic in the urinary bladder of the painted reed frog Hyperolius marmoratus marmoratus collected in southern Natal, South Africa. Parasite prevalence varied from 14.5 to 47.4% and the mean intensity from 1.0 to 1.1. The hamuli of the new species have a characteristic, partly fragmented appearance, a feature also seen in Polystoma batchvarivi, the only other polystome described from a species of the genus Hyperolius. The similarity between the two species, in spite of their geographical separation, is commented upon.     

Inhibition by vitamin A of cyst formation in Nyctotheroides puytoraci induced by injecting its host Bufo regularis with 20-methylcholanthrene

20-Methylcholanthrene induced cyst formation in Nyctotheroides puytoraci when injected into its host Bufo regularis. Presumably this hydrocarbon or its metabolites reaches the parasites in the recta of treated host animals and induces encystment. However, injection B. regularis with 0.5 mg of 20-methylcholanthrene + vitamin A palmitate (5,000 IU) inhibited the hydrocarbon-induced encystment of the parasites.     

滇蛙多盘吸虫精子发生的超微结构

利用透射电镜研究滇蛙多盘吸虫的精子发生。它属于典型的寄生扁形动物Ｃｅｒｃｏｍｅｒｉｄｅａ类型。本吸虫的特点是精子形成的早期,两根鞭毛轴丝相互生趣成９０℃,中央间体由三根带条带组成,不等长,中央带长于侧带;线粒体在融合前未观察到围核现象。     

Gill monogeneans of neotropical cichlid fish: diversity,phylogenetic relationships,and host-parasite cophylogenetic associations

Host-parasite coevolution is one of the main topics of the evolutionary biology of host-parasite associations. The majority of monogeneans parasitizing fish exhibit a high degree of host specificity. As a result, their evolutionary history might be intertwined with that of their fish hosts. The Cichlidae represent a diverse group of secondary freshwater fish with disjunctive distribution. Host-specific dactylogyrid monogeneans commonly parasitize cichlid fish. Their high diversity is associated with the main areas of cichlid distribution, i.e., Neotropical America and Africa. Nevertheless, the parasite fauna of cichlids from Neotropical America is still underexplored. A total of 31 cichlid species were examined for the presence of monogeneans, with 20 of them being parasitized. On these cichlids, 30 monogeneans belonging to the genera Gussevia, Trinidactylus, and Scadicleithrum were identified, 17 of them potentially representing new species for science. Phylogenetic analyses revealed three monophyletic groups of Neotropic cichlid monogeneans. Genus Gussevia was monophyletic, while Sciadicleithrum resulted polyphyletic. Sciedicleithrum from South America and Sciadicleithrum from Mexico represented two divergent lineages. The plesiomorphic Neotropical cichlid host group for dactylogyrid monogeneans was Cichlini, from which the representatives of other Neotropical cichlid tribes were colonised. Cophylogenetic analyses revealed a statistically significant cophylogenetic signal in the investigated host-parasite system, with host switch and duplication representing the main coevolutionary events for monogeneans parasitizing Neotropical cichlids. This scenario is in accordance with previous studies focussed on dactylogyridean monogeneans parasitizing freshwater fish in Europe and Africa.     

First monogenean flatworm from a microhylid frog host: Kankana, a new polystome genus from Madagascar

Kankana manampoka n. gen., n. sp. (Monogenea, Polystomatidae), is described from the urinary bladder of the narrow-mouthed frog Platypelis pollicaris. This is the first record of a polystome from the Microhylidae and the third polystome genus from Madagascar, next to Metapolystoma and Madapolystoma. The extensive uterus and presence of hamuli resemble Metapolystoma but the vitellarium confined to the lateral fields in Kankana is different. Madapolystoma also has an extensive uterus but contain only up to 32 advanced developed larvae. Based on the extensive uterus filling the body proper and the vitellarium confined to two lateral fields posterior in the body this new polystome resembles Eupolystoma known from Africa and India. However, unlike Eupolystoma, the gonads are in the middle of the body, vaginae are lacking and a pair of hamuli is present. A molecular phylogenetic analysis of concatenated 18S and 28S ribosomal RNA gene sequences supplemented by genetic distances inferred from 28S and COI sequences showed that this new genus is more related to Madapolystoma, a genus only reported from Madagascar, than to Eupolystoma known from Africa and India and Metapolystoma known from Africa and Madagascar.     

Morphology and molecular characterization of Polystoma goeldii n. sp. (Monogenea,Polystomatidae) parasite from the urinary bladder of Physalaemus ephippifer (Steindachner) (Anura,Leptodactylidae)

Polystomatid flatworms of amphibians are represented in the Neotropical realm by species of Mesopolystoma, Nanopolystoma, Parapseudopolystoma, Polystoma, Riojatrema and Wetapolystoma but only species of Polystoma are known from Brazil, namely Polystoma cuvieri, P. knoffi, P. lopezromani and P. travassosi. During a survey of monogeneans infecting amphibians in the north-eastern region of Pará State, the Cayenne Caecilian Typhlonectes compressicauda was found to be infected with Nanopolystoma tinsleyi and the Veined Tree Frog Trachycephalus typhonius was found to harbor Polystoma lopezromani. A yet unknown species of Polystoma was also encountered in the urinary bladder of the Steindachner's Dwarf Frog, Physalaemus ephippifer. This new species, which is the second species reported from Physalaemus spp., is described herein as Polystoma goeldii n. sp. and its life cycle is also illustrated. The new species can be distinguished from Polystoma spp. from other neotropical realm by a combination of characteristics, including hamuli morphology, outer/inner hamuli length ratio, haptor/total body length ratio, genital bulb/total body length ratio, genital spine number and COI molecular characters.     

Four species of Pseudorhabdosynochus (Monogenea: Diplectanidae) from the camouflage grouper Epinephelus polyphekadion (Perciformes: Serranidae) off New Caledonia

Gill diplectanid monogeneans from the camouflage grouper Epinephelus polyphekadion (Bleeker) collected in the coral reef lagoon of New Caledonia, South Pacific, comprise four species. Very few monogeneans were found in most fish examined. Pseudorhabdosynochus dionysos n. sp. has a sclerotised vagina with a robust trumpet, a robust primary canal and two chambers of similar size; it is close to P. bacchus Sigura, Chauvet & Justine, 2007. P. viscosus n. sp. has a sclerotised vagina with a robust trumpet, long primary canal with an extremely thin wall and two small chambers, and a male quadriloculate organ with a characteristic thickening at the extremity of its cone. P. crassus n. sp., the most abundant species, has a sclerotised vagina with a thin-walled trumpet, thin-walled primary canal which is always coiled anteriorly and two small chambers. P. huitoe Justine, 2007, P. manifestus Justine & Sigura, 2007 and P. crassus have very similar sclerotised vaginae; however, species of this 'huitoe complex' can be distinguished by measurements of the haptoral hard parts. A few diplectanid specimens found in a single specimen of E. polyphekadion were attributed to P. huitoe, a species originally described from E. maculatus (Bloch) and also rarely found in E. cyanopodus Richardson in New Caledonia; specimens from these three fish species are morphologically indistinguishable.     

Comparative microscopical study of the gall bladder mucosa.

The gall bladder from 6 Psammophis sibilans, 10 Bufo regularis and 10 Albino mice were extracted and prepared for microscopic examination. It was found that the mucosa of Psammophis sibilans consisted of ovoid and polygonal cells which were occasionally binucleated cells with darkly stained nuclei and occasionally pear-shaped cells with vesicular nuclei and fine processes. These cells were arranged in three layers. Apossible explanation for the different types of cells encountered and their arrangement was given. The gall bladder mucosa of Bufo regularis and Albino mouse were thrown into folds covered with simple columnar epithelium. However, the epithelium of the frog was higher than that of the mouse, with the nuclei situated midway between basement membrane and the lumen. Vacuolated cells were detected in the gall bladder mucosa of the mouse. The significance of the mucosal folds was discussed.     

Differences between inter- and intraspecific architectonic adaptations to pharyngeal mollusc crushing in cichlid fishes

Because fish heads are densely packed with muscles, ligaments, skeletal elements and other structures, transformations in one structure may influence surrounding structures. Transformations occur during phylogeny, ontogeny and as environmentally induced alterations, i.e. phenotypic plasticity. We describe differences in intra- and interspecific transformations of the pharyngeal jaw apparatus of haplochromine cichlids. Using multivariate clustering techniques we trace possible correlations in transformations of anatomical characters of the pharyngeal jaws and surrounding structures. The intraspecific transformation analysis is based on two environmentally induced morphs of Astatoreochromis alluaudi : a molluscivorous morph with a hypertrophied pharyngeal jaw apparatus and an insectivorous one with a non-hypertrophied apparatus. For the interspecific analysis five other haplochromine species from Lake Victoria with diets ranging from insects to molluscs were investigated. Although ranges in diet are the same, the anatomical ranges differ between A. alluaudi and the species cline. Besides similarities in anatomical changes of the pharyngeal jaw apparatus in the intra- and interspecific cline, differences were also observed. Apparently there are among haplochromines multiple pathways to achieve similar performance. In A. alluaudi architectonic and intrinsic plasticity constraints limit the adaptability of the pharyngeal jaw apparatus. In the species cline, these constraints have been overcome by genetical adaptation.     

Multilocus species tree analyses resolve the radiation of the widespread Bufo bufo species group (Anura, Bufonidae)

New analytical methods are improving our ability to reconstruct robust species trees from multilocus datasets, despite difficulties in phylogenetic reconstruction associated with recent, rapid divergence, incomplete lineage sorting and/or introgression. In this study, we applied these methods to resolve the radiation of toads in the Bufo bufo (Anura, Bufonidae) species group, ranging from the Iberian Peninsula and North Africa to Siberia, based on sequences from two mitochondrial and four nuclear DNA regions (3490 base pairs). We obtained a fully-resolved topology, with the recently described Bufo eichwaldi from the Talysh Mountains in south Azerbaijan and Iran as the sister taxon to a clade including: (1) north African, Iberian, and most French populations, referred herein to Bufo spinosus based on the implied inclusion of populations from its type locality and (2) a second clade, sister to B. spinosus, including two sister subclades: one with all samples of Bufo verrucosissimus from the Caucasus and another one with samples of B. bufo from northern France to Russia, including the Apennine and Balkan peninsulas and most of Anatolia. Coalescent-based estimations of time to most recent common ancestors for each species and selected subclades allowed historical reconstruction of the diversification of the species group in the context of Mediterranean paleogeography and indicated a long evolutionary history in this region. Finally, we used our data to delimit the ranges of the four species, particularly the more widespread and historically confused B. spinosus and B. bufo, and identify potential contact zones, some of which show striking parallels with other co-distributed species.     

Inbreeding depression in two seed-feeding beetles, Callosobruchus maculatus and Stator limbatus (Coleoptera: Chrysomelidae)

Inbreeding depression is well documented in insects but the degree to which inbreeding depression varies among populations within species, and among traits within populations, is poorly studied in insects other than Drosophila. Inbreeding depression was examined in two long-term laboratory colonies of the seed beetle, Callosobruchus maculatus (Fabricius), which are used frequently as models for experiments in ecology, evolution and behaviour. Inbreeding depression in these laboratory colonies are compared with one recently field-collected population of a different seed beetle, Stator limbatus Horn. Inbreeding reduced embryogenesis, egg hatch and larval survival in both species, such that eggs produced by sib matings were >17% less likely to produce an adult offspring. Inbred larvae also took 4-6% longer to develop to emergence in both species. Inbreeding depression varied among the measured traits but did not differ between the two populations of C. maculatus for any trait, despite the large geographic distance between source populations (western Africa vs. southern India). Inbreeding depression was similar in magnitude between C. maculatus and S. limbatus. This study demonstrates that these laboratory populations of C. maculatus harbour substantial genetic loads, similar to the genetic load of populations of S. limbatus recently collected from the field.     

滇蛙多盘吸虫精子超微结构

利用透射电镜研究多盘吸虫精子的形态和结构。它属于单殖类中Ｐｏｌｙｏｐｉｓｔｈｏｃｏｔｙｌｅａ类型。精子细线形,含两根融入精子内的９＋１型轴丝,细长的细胞核,纤细的线粒体和微管。在第二根轴丝出现前的区段无微管的分布,具两根轴丝的主干段轴丝外无外侧管;细胞核消失后微管呈整圈排列;精子后部含糖原颗粒,背腹面中央向髓布内凹呈哑铃状并两侧向外突出形成波动膜;由精细胞则来的区段具外饰物。     

我国内陆水体单殖吸虫的宿主特异性分析

报道我国内陆水体单殖吸虫的宿主特异性的研究。研究表明单殖吸虫有很强的宿主特异性,但不同种类的特异的有很大的判别。从目前已有的记录来看,６０％以上的单殖吸虫中只有１种宿主,约７５％的单殖吸虫的宿主仅为１属,超过９７％的单殖吸虫的宿主在１科之内。没有１种单殖吸虫可生于超过３个料的宿主中,但大钩指环虫可在２３属３３种宿主中寄生。在科和发放持异性水平,以锚首虫科和指环虫科单殖吸虫的特异性较强,三代虫科次之