Multiplex 16S rRNA haplotype-specific PCR, a rapid and convenient method for fish species identification: an application to West African Clupeiform larvae

A multiplex haplotype-specific polymerase chain reaction (MHS-PCR) method was developed, which identified seven Clupeiform species living in the tropical Eastern Atlantic region: Sardinella aurita, Sardinella maderensis, Ethmalosa fimbriata, Sardina pilchardus, Engraulis encrasicholus, Pellonula leonensis and Ilisha africana. 16S rRNA fragments were amplified using a species-specific set of primers, yielding species-specific size fragments, and then separated using agarose gel electrophoresis, enabling direct visual identification of targeted species. This method provides an accurate, easy and rapid tool for identifying species within large Clupeiform samples. It is suitable for investigations on early Clupeiform stages, species and identification in fishery management in the tropical Eastern Atlantic area.     

The structure of anchovy outer retinae (Engraulididae, Clupeiformes) - a comparative light- and electron-microscopic study using museum-stored material

The outer retinal architecture of Engraulididae is uncommon among vertebrates. In some anchovies, e.g., Anchoa, two cone types are arranged alternating in long photoreceptor chains, i.e., polycones. The cones have radially oriented outer segment lamellae in close contact with a complex guanine tapetum, most probably subserving polarization contrast vision. To clarify the distribution of the aberrant polycone architecture within the Engraulididae and to provide indications about polycone evolution, the outer retina morphology of 16 clupeoid species was investigated by light and electron microscopy, predominantly using museum-stored material. The outgroup representatives of four clupeid subfamilies (Clupeonella cultriventris, Dorosoma cepedianum, Ethmalosa fimbriata, Pellonula leonensis) show a row pattern of double cones, partially with single cones at defined positions and a pigment epithelium with lobopodial protrusions containing melanin. The pristigasterid Ilisha africana has double rows of single cones lying between linear curtains of pigment epithelium processes filled with minute crystallites and melanin concentrated near their vitreal tips. Within the Engraulididae, two main architectures are found: Coilia nasus and Thryssa setirostris have linear multiple cones or polycones separated by long pigment epithelium barriers containing tapetal crystallites and melanin in the tips (also found in Setipinna taty), whereas Anchoviella alleni, Encrasicholina heteroloba, Engraulis encrasicolus, Engraulis mordax, Lycengraulis batesii, and Stolephorus indicus exhibit the typical polycone architecture. Cetengraulis mysticetus and Lycothrissa crocodilus show cone patterns and pigment epithelium morphology differing from the other anchovy species. The sets of characters are compared, corroborated with the previous knowledge on clupeoid retinae and discussed in terms of functional morphology and visual ecology. A scenario on polycone evolution is developed that may serve as an aid for the reconstruction of engraulidid phylogeny. Furthermore, this study demonstrates the suitability of museum material for morphological studies, even at the electron microscopic level.     

The reproductive biology of Pellonula afzeliusi Johnels,and Sierrathrissa leonensis thys Audenaerde in Lake Kainji,Nigeria

Aspects of the reproductive biology of the two species of clupeids known to occur in Lake Kainji, Pellonula afzeliusi Johnels and Sierrathrissa leonensis Thys Van Den Audenaerde were studied for a period of twenty eight months. Mature individuals were found in both species all the year round but with a prolonged peak of activity from December to May, more distinct in P. afzeliusi than in S. leonensis.There were no significant length differences between mature males and females in both species. The smallest mature individuals were 28 mm standard length female; 27 mm standard length male in P. afzeliusi, while the corresponding figures for S. leonensis were 18 mm standard length female and 19 mm standard length male.No sexual dimorphism was observed. There was a simple correlation between fecundity and fish standard body length as well as weight in both species.     

Studies on the excystment of trematode metacercariae in vitro.

Excystment of metacercariae of Parorchis acanthus, Posthodiplostomoides leonensis, Posthodiplostomum sp. and Clinostomum tilapiae was found to be an active process initiated by the metacercariae when the correct physico-chemical conditions exist in the environment; but in the excystment of Posthodiplostomum nanum the metacercaria seems not to take part in the process. Temperature, osmotic pressure, pH and eH are factors which influence all the five species studied. Fresh ox bile was found not to be an absolute necessity for the excystment of the five species studied but its presence in the medium increased the rate of excystment of three species.     

Excystation of five digenean metacercariae in vitro and in the peritoneal cavity of the mouse

Excystation of metacercariae of Parorchis acanthus, Posthodiplostomum nanum, Posthodiplostomum sp., Posthodiplostomoides leonensis and Clinostomum tilapiae in mammalian saline at 39 degrees C was similar to their excystation in mouse peritoneal cavity. Extrinsic enzymes were not needed for C. tilapiae excystation but were an absolute necessity for P. acanthus excystation. Extrinsic pepsin enhanced P. nanum excystation but was not obligatory. Acid pepsin was an absolute requirement for Posthodiplostomum sp. and Posthodiplostomoides leonensis excystation. It was noted that acid saline could not be substituted for acid pepsin and that there was diversity in the condition involved in digenean excystation.     

Phylogeographic patterns in populations of the black-chinned tilapia complex (Teleostei,Cichlidae) from coastal areas in West Africa: support for the refuge zone theory

Within the tilapiines, a major African cichlid lineage, quite limited genetic informations are available on phylogenetic interrelationships and phylogeographical patterns at both macro- and micro-evolutionary scales. The present study examines the genetic diversity of the black-chinned tilapia complex from coastal areas in West Africa (Senegal to Congo-Brazzaville) based on mtDNA control region sequences. Phylogenetic inferences provide support for the monophyly of both taxa involved. It is suggested that Sarotherodon melanotheron and Sarotherodon nigripinnis have diverged probably since the early Pleistocene. The occurrence of the main intraspecific lineages could be dated back to about 500,000-900,000 years. Our data suggest that West African Pleistocene refuge zones may have strongly influenced the degree and level of genetic differentiation among populations and thus the observed phylogeographic patterns. The spatial distribution of mtDNA lineages and the centres of intrapopulational genetic diversity clearly correspond to proposed lowland refuge zones and core areas of Central Africa. Moreover, this study genetically supports the existence of Sarotherodon melanotheron leonensis for the first time and further confirms the validity of the most recently introduced subspecies, e.g., Sarotherodon nigripinnis dolloi.     

The organization of interphase chromatin in drosophilidae

Cytological evidence is presented which shows that for Drosophila virilis and Samoaia leonensis at least, each satellite DNA is condensed into a distinct heterochromatic mass during interphase. This is seen as just one example of a general phenomenon in which chromatin containing a particular DNA sequence binds to other chromatin containing the same sequence. It is proposed that DNA sequence specific proteins can account for this phenomenon.     

Phylogeny of Selected Maxillopodan and Other Crustacean Taxa Based on 18S Ribosomal Nucleotide Sequences: A Preliminary Analysis

Relationships among the following taxa were examined based on 18S ribosomal RNA and DNA nucleotide sequences: Branchiopoda (Branchinecta packardi), Ostracoda (two podocopid species), Branchiura (Argulus nobilis), Pentastomida (Porocephalus crotali), Copepoda (Calanus pacificus, Macrocyclops albidus), Thoracica (Balanus eburneus, Calantica villosca), Acrothoracica (Trypetesa lampas), and Decapoda (Procambarus leonensis, Callinectes sapidus). Phylogenetic relationships were inferred by maximum parsimony and results evaluated by bootstrapping. With Branchinecta packardi as an outgroup, the following tree was inferred: the first branch leads to a node that joins a pentastome, a branchiuran and the two ostracodes; the second branch leads to a node that has the two copepods as a sister taxon to a clade that includes the acrothoracican and thoracicans; the last branch leads to a node joining the two decapods. The analyses suggest: (1) acrothoracicans diverged very early from the cirripede line and are not derived from a lepadomorph-like ancestor; (2) branchiurans are not related to either copepods or thecostracans but are closely allied with pentastomes; and (3) the Maxillopoda, broadly defined, is not a monophyletic taxon. These results differ from previous studies of these groups.     

The food and feeding habits of Eutropius niloticus (Ruppell), family Schilbeidae (Osteichthyes: Siluriformes) in Lake Kainji,Nigeria

Studies on the food and feeding habits of Eutropius niloticus in Lake Kainji, Nigeria revealed that the fish is omnivorous but fed more on insects than on any other food. The most important insects were Povilla adusta Navas nymphs and imagos and adults of the common grasshoppers. Fish, particularly the clupeids Sierrathrissa leonensis Thys Van den Audernaerde and Pellonula afzeliusi Johnels, also formed an important part of the diet. There was seasonal variation of organisms selected for food and there were variations in the dietary items selected by different age groups. Accumulation of fat was found to correlate with seasonal feeding intensity. E. niloticus serves as food for other fishes.     

Biology of the small pelagic fishes in the new Volta Lake in Ghana

Summary A number of small pelagic fish species were found in the new Volta Lake in Ghana. Three, the clupeids Cynothrissa mento and Pellonula afzeliusi, and the schilbeid Physailia pellucida, rapidly became dominant, and these together with another schilbeid Siluranodon auritus and the clupeid-like cyprinid Barilius niloticus are here considered. Pellonula and Physailia eat chiefly invertebrates; the former especially at the surface, the latter in mid-depths or at the bottom. Zooplankton is important to all sizes of Physailia, but only to juveniles of Pellonula. Barilius feeds almost entirely on adventitous forms taken at the surface and Siluranodon on algae and rotifers. Cynothrissa, the largest fish studied, feeds mainly on fish. The food range taken is thus wide, and differences in diet are discussed in terms of the probable future of each species in the new lake as it matures.This paper is taken from a thesis accepted for the M. Sc. Degree of the University of Dublin.     

On the biology of an estuarine population of the clupeidPellonula afzeliusi (Johnels) in Lagos Lagoon,Nigeria

The length-weight relationship, condition factor, fecundity and food and feeding habits ofPellonula afzeliusi (Johnels) in Lagos Lagoon were investigated. The standard length of 1832 specimens examined ranged between 3.5 and 9.4 cm (total length 3.9 cm–11.4 cm) and was larger than reported elsewhere in West African man-made lakes. Specimens exhibited positive allometric growth. Their condition factor increased with individual size. Females had a slightly higher condition factor than males. The number of mature ova per female ranged between 330 and 33,649. There was a higher correlation between fecundity and weight than between fecundity and length. The major food items were crustaceans, insects and fishes. There were distinct differences in the feeding habits in relation to size. Variations were observed in the monthly analysis of food items.     

Virion RNA species of the arenaviruses Pichinde, Tacaribe, and Tamiami.

The principal RNA species isolated from labeled preparations of the arenavirus Pichinde usually include a large viral RNA species L (apparent molecular weight = 3.2 X 10(6)), and a smaller viral RNA species S (apparent molecular weight = 1.6 X 10(6)). In addition, either little or considerable quantities of 28S rRNA as well as 18S rRNA can also be obtained in virus extracts, depending on the virus stock and growth conditions used to generate virus preparations. Similar RNA species have been identified in RNA extracted from Tacaribe and Tamiami arenavirus preparations. Oligonucleotide fingerprint analyses have confirmed the host ribosomal origin of the 28S and 18S species. Such analyses have also indicated that the Pichinde viral L and S RNA species each contain unique nucleotide sequences. Viral RNA preparations isolated by conventional phenol-sodium dodecyl sulfate extraction often have much of their L and S RNA species in the form of aggregates as visualized by either electron microscopy or oligonucleotide fingerprinting of material recovered from the top of gels (run by using undenatured RNA preparations). Circular and linear RNA forms have also been seen in electron micrographs of undenatured RNA preparations, although denatured viral RNA preparations have yielded mostly linear RNA species with few RNA aggregates or circular forms.     

Dynamics of drift, gene flow, and selection during speciation in Silene

The mechanics of speciation with gene flow are still unclear. Disparity among genes in population differentiation (F(ST)) between diverging species is often interpreted as evidence for semipermeable species boundaries, with selection preventing "key" genes from introgressing despite ongoing gene flow. However, F(ST) can remain high before it reaches equilibrium between the lineage sorting of species divergence and the homogenizing effects of gene flow (via secondary contact). Thus, when interpreting F(ST), the dynamics of drift, gene flow, and selection need to be taken into account. We illustrate this view with a multigenic analyses of gene flow and selection in three closely related Silene species, S. latifolia, S. dioica, and S. diclinis. We report that although S. diclinis appears to have evolved in allopatry, isolation with (bidirectional) gene flow between S. latifolia and S. dioica is likely, perhaps as a result of parapatric speciation followed by more extensive sympatry. Interestingly, we detected the signatures of apparently independent instances of positive selection at the same locus in S. latifolia and S. dioica. Despite gene flow between the species, the adaptive alleles have not crossed the species boundary, suggesting that this gene has independently undergone species-specific (diversifying or parallel) selection.     

Steinernema cholashanense n. sp. (Rhabditida, Steinernematidae) a new species of entomopathogenic nematode from the province of Sichuan, Chola Shan Mountains, China

During a random survey of entomopathogenic nematodes in the provinces of Sichuan and Gansu (eastern Tibet) in 2004, soil samples from several sites were collected and tested for the incidence of entomopathogenic nematodes. A new species was collected in this survey and it is described herein as Steinernema cholashanense n. sp. Steinernema cholashanense n. sp. is characterized by morphology and morphometry of the IJ and male. For the IJ, the new species can be recognized by the average body length 843 microm, esophagus length 125 microm, H%=39% and E%=81%. The lateral field pattern is 2, 5, 7, 4, 2. The male of the first generation is characterized by spicule shape and length and especially with prominent velum and the presence of a mucron on both generations. The average body length of the IJ of S. cholashanense n. sp. (843 microm) is shorter than that of S. oregonense (980 microm), S. kraussei (951 microm) and S. litorale (909 microm), similar to that of S. feltiae (849 microm), but longer than that of S. weiseri (740 microm), S. jollietti (711 microm) and S. hebeiense (658 microm). Esophagus length of the new species (125 microm) is closer to that of S. jollieti (123 microm) but longer than that of S. weiseri (113 microm) and shorter than that of S. oregonense (132 microm), S. kraussei (134 microm) and S. feltiae (136 microm). E% of the new species (81) is similar to that of S. kraussei (80), but smaller than that of S. jollieti (88), S. weiseri (95), S. oregonense (100) and S. feltiae (119). Spicule head length of the new species is almost the same as its width, this character is similar to that of S. kraussei but it is different from this species by its prominent velum. The new species can be recognized further by characteristics of sequences of ITS and D2D3 regions and cross hybridization with closely related species, S. feltiae, S. kraussei and S. oregonense.     

Sequence diversity, reproductive isolation and species concepts in Saccharomyces

Using the biological species definition, yeasts of the genus Saccharomyces sensu stricto comprise six species and one natural hybrid. Previous work has shown that reproductive isolation between the species is due primarily to sequence divergence acted upon by the mismatch repair system and not due to major gene differences or chromosomal rearrangements. Sequence divergence through mismatch repair has also been shown to cause partial reproductive isolation among populations within a species. We have surveyed sequence variation in populations of Saccharomyces sensu stricto yeasts and measured meiotic sterility in hybrids. This allows us to determine the divergence necessary to produce the reproductive isolation seen among species. Rather than a sharp transition from fertility to sterility, which may have been expected, we find a smooth monotonic relationship between diversity and reproductive isolation, even as far as the well-accepted designations of S. paradoxus and S. cerevisiae as distinct species. Furthermore, we show that one species of Saccharomyces--S. cariocanus--differs from a population of S. paradoxus by four translocations, but not by sequence. There is molecular evidence of recent introgression from S. cerevisiae into the European population of S. paradoxus, supporting the idea that in nature the boundary between these species is fuzzy.     

Affinities between Asian non-human Schistosoma species,the S. indicum group,and the African human schistosomes

Schistosoma species have traditionally been arranged in groups based on egg morphology, geographical origins, and the genus or family of snail intermediate host. One of these groups is the 'S. indicum group' comprising species from Asia that use pulmonate snails as intermediate hosts. DNA sequences were obtained from the four members of this group (S. indicum, S. spindale, S. nasale and S. incognitum) to provide information concerning their phylogenetic relationships with other Asian and African species and species groups. The sequences came from the second internal transcribed spacer (ITS2) of the ribosomal gene repeat, part of the 28S ribosomal RNA gene (28S), and part of the mitochondrial cytochrome c oxidase subunit 1 (CO1) gene. Tree analyses using both distance and parsimony methods showed the S. indicum group not to be monophyletic. Schistosoma indicum, S. spindale and S. nasale were clustered among African schistosomes, while S. incognitum was placed as sister to the African species (using ITS2 and 28S nucleotide sequences and CO1 amino acid sequences), or as sister to all other species of Schistosoma (CO1 nucleotide sequences). Based on the present molecular data, a scenario for the evolution of the S. indicum group is discussed.     

Pollination systems, biogeography, and divergence times of three allopatric species of Schisandra in North America, China, and Japan

This study analyses the pollination systems and biogeography of three allopatdc species of Schisandra (Section Euschisandra) consisting of S.glabra (North America),S.bicolor (China),and S.repanda (Japan); the clade is delimited in a phylogenetic tree of Schisandraceae constructed with nuclear and plastid genes.The male and female flowers of these species have similar floral structures,but exhibit different pollination systems.At the base of the clade,S.glabra is pollinated by a wide variety of beetles and flies in a generalist pollination system that also includes floral heat and the use of male and female flowers as brood sites for insects.In Asia,however,S.bicolor and S.repanda are pollinated exclusively by one or two different species of gall midges (Resseliella spp.) in a specialist pollination system.In this system only female,pollen-eating gall midges pollinate the flowers and breed on nearby spiderwebs.The gall midge pollination system is specialized and derived from the generalist system in S.glabra,and basal in the clade.Pollen is the main floral resource,and we hypothesize it is exploited to enrich eggs,and as a result species of gall midges could increase reproductive fitness by feeding on a single dependable food source.Subsequently the life cycles of the plants and insects evolved into a tight association in old stable plant communities in the Sino-Japanese flora.Divergence times for the plant species are presented and correlated with past distributions and migration routes.     

深圳市大南山地区铁榄群落研究

对深圳市大南山地区的主要植被进行调查研究。结果表明,铁榄(Sinosideroxylon wightianum)常生长在瘠薄坡地、陡峭沟谷中,是该地区最重要的天然林群落,常组成单优势群落,呈斑块状分布,也是该地区存留的少数次生常绿阔叶林群落之一;大南山地区的铁榄群落有10个主要类型,铁榄是常绿阔叶林植被的主要特征种;大南山地区铁榄群落结构简单,物种分布不均匀,物种多样性指数差异较大;铁榄以Ⅱ级小树为主,兼有Ⅰ级苗木和Ⅲ级壮树,表明铁榄在群落中的优势度仍将保持较长一段时间;铁榄与漆树等物种间的种间联结关系薄弱,表明群落处于群落演替的早期发育阶段。