A histological and scanning electron microscope study of the formation of the wall plates in Chthamalus depressus (Poli)

Because of the small amount of pigmentation the difficulties associated with earlier histological work on Chthamalus stellatus (Poli) are absent in extreme hypobiotic C. depressus (Poli). The formation of the wall plates has been investigated by normal histological techniques and supported by scanning electron microscopy. The wall plates are covered by epicuticle which in its continuity resembles that of Acasta spongites; the epicuticle is strongly ribbed and the plate has more or less regularly spaced, volcano-like outgrowths. The various cell-types and the part they play in the expansion of the basis and growth of the wall plate are described. A series of ‘bridging’ cells are present and lie between the lower part of the last completely formed ‘scale’ and the basal cells. Growth is considered from the evolutionary point of view and in relation to the forms assumed under natural conditions, in particular to the capacity of the animal to adapt to changes in the rock surface.     

Microphallidae (Digenea) d'Afrique du Sud,Parasites de Charadrii (Aves). Seconde Note

Les auteurs apportent de nouvelles informations géographiques et diagnostiques concernant plusieurs microphallidés (Trematoda: Digenea) récoltés chez des Charadrii (Aves) des c^otes de Namibie (Afrique du Sud): Maritrema eroliae Yamaguti, 1939 de Charadrius marginatus Vieillot; Odhneria odhneri Travassos, 1921 de Arenaria interpres L.; Microphallus bilobatus Cable, Connor &; Balling, 1960 de C. marginatus; et Levinseniella propinqua Jägerskiöld, 1907 de C. marginatus et A. interpres. Ces espèces représentent une localisation géographique nouvelle et de nouveaux h^otes. Le phallus (organe copulateur m^ale) de M. bilobatus de Namibie et celui du type des Caraïbes sont comparés et illustrés. Les atriums génitaux de L. propinqua récoltés en Namibie ainsi qu'à Marcus Island (southwest Cape Province, Afrique du Sud) sont illustrés et discutés; l'espèce parait cosmopolite. Abstract The authors present new geographical and diagnostic information for microphallids (Trematoda: Digenea) from the coast of Namibia (southern Africa): Maritrema eroliae Yamaguti, 1939 from Charadrius marginatus Vieillot; Odhneria odhneri Travassos, 1921 from Arenaria interpres L.; Microphallus bilobatus Cable, Connor &; Balling, 1960 from C. marginatus; and Levinseniella propinqua Jägerskiöld, 1907 from C. marginatus and A. interpres. These are new geographical and host records. The position and variability of the phallus (male copulatory organ) in M. bilobatus from Namibia and in the type-species from the Caribbean Sea are compared and illustrated. The genital atria of L. propinqua from Namibia and from Marcus Island (southwest Cape Province, Southern Africa) are illustrated and compared. This species appears to be cosmopolitan.     

Biological evaluation of the systematic validity of the African Argas (Persicargas) Arboreus and the Asian-Australian A. (P.) Robertsi (Ixodoidea: Argasidae)

Khalil G. M., Hoogstraal H. and Oliver J. H. Jr. 1980. Biological evaluation of the systematic validity of the African Argas (Persicargas) arboreus and the Asian-Australian A.(P.)robertsi (Ixodoidea: Argasidae). International journal for Parasitology10: 253–259. The closely related A. (P.)arboreus (Ethiopian Faunal Region) and A. (P.) robertsi (Oriental and Australian Faunal Regions) interbreed readily and produce partially fertile progeny. Intrahybrid crosses are much less productive than P₁ homogamic crosses. Results of backcrosses indicate that the progeny of the robertsi ♂ × arboreus ♀ cross are less fertile than the progeny of the reciprocal cross. The fertility of F₁ hybrid progeny is lower than that of the P₁ pure species as expressed in lower female fecundity, egg hatch, and/or viability of immature stages. These 2 species probably originated from a common ancestor and geographic isolation caused genetic incompatibility.     

Hybridization of Australian Haemonchus placei (place, 1893), Haemonchus contortus cayugensis (Das & Whitlock, 1960) and Haemonchus contortus (Rudolphi, 1803) from Louisiana

There were distinct ecotypie differences in the ability to develop to third stage larvae at a constant 11 or 13°C for two weeks. H. conforms cayugensis could develop at both 11 and 13°C; H. contortus from Louisiana could develop at 13°C but not 11°C and H. placei could not complete development at either temperature. Eggs produced from the first cross between ecotypes had the same cold tolerance as their maternal parent. F₁ eggs were intermediate between parental ecotypes, while F₂ eggs, when fertile, developed at the same temperature as the most cold-tolerant parent in the first cross. The H. placei knobbed vulvar morph type was dominant to H. contortus cayugensis smooth morph type and recessive to the H. contortus (Louisiana) smooth morph type. Knobbed morph type of H. contortus (Louisiana) was dominant to smooth of H. contortus cayugensis. Dominance of morph type was enhanced by a matroclinous effect in the between species matings but not in the within species matings. No fertile eggs were produced by the F1 of the mating between female H. placei × male H. contortus (Louisiana) or by the F₂ of the reciprocal mating. Reciprocal matings between H. placei × H. contortus cayugensis produced an F₁ and F₂ that had reduced fertility but were not completely sterile. No decrease in fertility was observed in the F₂ and F₁ from the H. contortus cayugensis × H. contortus(Louisiana)matings. Cytological studies revealed several kinds of meiotic disturbances in the between species F₁ and F₂. The most frequent were aneuploidy, failure of chromosome pairing, and pairing between non-homologous chromosomes. Males were more seriously affected than females by meiotic disturbances. The H. placei X chromosome appeared to be selected against by the hybrid genetic background. Within species hybrids showed no meiotic disturbances and the percentage of male offspring from these crosses was the same as for female offspring.     

A molecular analysis of some Eastern Atlantic grouper from the Epinephelus and Mycteroperca genus

Mitochondrial cytochrome b (397 bp) and 16S rDNA (516 bp) sequences analysis was used to investigate the phylogenetic relationships among some Eastern Atlantic Epinephelinae species. Six species of Epinephelus (E. aeneus, E. caninus, E. costae, E. haifensis, E. marginatus and E. tauvina) and two species of Mycteroperca (M. rubra and M. fusca) were analysed. Neighbour-joining and maximum-parsimony analysis support the paraphyletic grouping of the Epinephelus and Mycteroperca analysed. The maximum pairwise nucleotide divergence value in cyt b among all taxa was 0.196 between E. aeneus and E. marginatus and the minimum value was 0.006 between E. costae and M. rubra. Meanwhile, in 16S sequence analysis, the maximum value is 0.093 between E. aeneus and E. tauvina and the minimum value is 0.011 between E. marginatus and M. rubra. Molecular clock estimates for the species suggest a divergence time of 20-24 mya, which coincides with the Miocene period. A molecular analysis was also conducted, using other Epinephelinae sequences from GenBank in order to improve our understanding of the phyletic status of the Epinephelus and Mycteroperca species analysed.     

The morphology of the larval stages of three species (Arrenurus mediorotundatus Thor, 1898; Arrenurus conicus Piers 1894; and Arrenurus cylindratus Piers, 1896) of the Megaluracarus subgenus (Acari: Hydrachnidia: Arrenuridae)

The morphology of the larval stages of three species (Arrenurus mediorotundatus, Arrenurus conicus and Arrenurus cylindratus) belonging to the Megaluracarus subgenus is described. Particular attention is paid to the common characters of the three species in which they differ from the other species belonging to the other subgenera. These are: the shape of the excretory pore plate, the shape of the dorsal plate, the length of coxal plate II medial margin and the C1-CpI, C4-CpIII distances. Other characters differ between the three species also. These are: the shape of the dorsal plate, the shape of coxal plate III, the shape of excretory pore plate and the presence or absence of secondary setae of the IFe3 setae.     

Morphology of Hydrozetes confervae () and H. parisiensis (Acari: Oribatida: Hydrozetidae), and keys to European species of Hydrozetes Berlese, 1902

Hydrozetes confervae [Schrank, F.P., 1781. Enumeratio insectorum Austriae indigenorum. Augustae Vindeliciorum, 1-552] and Hydrozetes parisiensis [Grandjean, F., 1948. Sur les Hydrozetes (Acariens) de l’Europe occidentale. Bull. Mus. Nat. Hist. nat., Paris, ser. 2(20), 328-335] are common European species of the aquatic oribatid mite family Hydrozetidae, but their external morphology is incompletely known. We describe and illustrate their larvae for the first time, and the nymphs of both species are redescribed and illustrated. Juveniles of the two species differ mainly by the position of gastronotal seta lm in relation to the opisthonotal gland opening (gla) and by the number of extremely long posterior hysterosomal setae in nymphs. In H. confervae seta lm inserts anteromedial to gla, but posterior to gla in H. parisiensis; nymphs of H. confervae have three pairs of long hysterosomal setae, while H. parisiensis has a larger, but variable number. Separate identification keys for the larvae, nymphs and adults of all known European species of Hydrozetes Berlese, 1902, are presented.     

Two new subspecies of Bacillus thuringiensis isolated in Japan: Bacillus thuringiensis subsp. kumamotoensis (serotype 18) and Bacillus thuringiensis subsp. tochigiensis (serotype 19)

Bacteriological and serological characteristics of three Bacillus thuringiensis isolates obtained in Japan were investigated. They formed typical rhomboidal parasporal inclusions but flagellar (H) antigens of these isolates were different from those of the known 17 H serotypes of B. thuringiensis. The three isolates were divided into two new serotypes (serotypes 18 and 19). The serotype 18 isolate (3–71) produced thermostable exotoxin and the inclusions of this isolate were toxic to larvae of the silkworm, Bombyx mori, but nontoxic to larvae of the mosquito, Aedes aegypti. The other isolate (119-72) belonging to serotype 18 produced inclusions nontoxic to larvae of B. mori and A. aegypti and did not produce thermostable exotoxin. However, other bacteriological properties of the isolate 119-72 were similar to those of the isolate 3–71. The serotype 19 isolate (117-72) produced inclusions nontoxic to larvae of B. mori and A. aegypti and did not produce thermostable exotoxin. Acid production from saccharose and the production of brownish purple pigment were observed in the two serotype 18 isolates, while neither of them was observed in the serotype 19 isolate. In other 29 biochemical properties tested, there was no difference among the three isolates. Based on these characteristics, the following two subspecies names are proposed: Bacillus thuringiensis subsp. kumamotoensis (serotype 18) for the type strain 3–71 and Bacillus thuringiensis subsp. tochigiensis (serotype 19) for the type strain 117-72.     

Megagametogenesis in Halophila johnsonii, a threatened seagrass with no known seeds,and the seed-producing Halophila decipiens (Hydrocharitaceae)

Megagametogenesis, the development of a megaspore into an embryo sac, has been identified in the seagrass Halophila johnsonii, a threatened species with no known sexual reproduction or seeds. Megagametogenesis in H. johnsonii was compared with megagametophyte development in Halophila decipiens, a related species known to readily produce viable seeds. In both species, ovules were structurally similar, megaspore mother cells were seen in premeiotic ovules, and linear tetrads and megagametophytes with two to eight nuclei were present in postmeiotic ovules. However, H. decipiens postmeiotic ovules had a chalazal pouch that was absent in the postmeiotic ovules of H. johnsonii. Late-stage H. decipiens ovules also contained embryos, indicating that they had been fertilized, whereas all late-stage H. johnsonii ovules were degrading and showed no signs of fertilization. These observations suggest that meiosis does occur in H. johnsonii megasporocytes, leading to the formation of viable megagametophytes and egg cells that could be fertilized if pollination occurred. Thus, the lack of seed set is due to a lack of pollination rather than any loss of capacity to produce seeds in this species.     

Hybridization studies of Haemonchus contortus (rudolphi, 1803) and H. placei (place, 1893) (Nematoda: Trichostrongylidae)

Hybridization experiments between Haemonchus contortus from sheep and H. placei from cattle indicate that the H. placei vulvar morph type and the inability of the eggs to hatch and develop at 11°C were inherited as dominant traits in the hybrids. The size of the third stage larvae was similar to H. placei in hybrids from the mating of male H. placei × female H. contortus while larvae from the reciprocal mating were intermediate in size. Hybrids produced by the mating of thiabendazole resistant female H. contortus × non-resistant male H. placei were also resistant. The F₁ males of the mating between male H. contortus and female H. placei were sterile. Male sterility did not occur in the reciprocal cross until the F₂ generation. Female hybrids from these generations had a low level of fertility when backcrossed to males of either parent species. Cytological studies of the hybrid males indicated that sterility was due to several kinds of meiotic disturbance and that spermatogenesis stopped during metaphase I. The chromosomes in eggs of unfertilized females did not undergo meiosis and polar body formation; instead they increased in number by a process of endomitosis. Unfertilized eggs in the faeces were characterized by uneven cytoplasmic division and abnormal shape. It is proposed that hybrid sterility can lead to the local eradication of one species of Haemonchus by the other. Furthermore, the outcome of which species is eradicated can be influenced by such variables as initial population sizes, climate, and the local ratio of sheep to cattle.     

Genetic structure of Phlebotomus (Larroussius) ariasi populations, the vector of Leishmania infantum in the western Mediterranean: Epidemiological implications

In recent years there has been growing interest in analyzing the geographical variations between populations of different Phlebotomus spp. by comparing the sequences of various genes. However, little is known about the genetic structure of Phlebotomus ariasi. In this study, we were able to sequence a fragment of the mitochondrial Cyt b gene in 133 sandflies morphologically identified as P. ariasi and proceeding from a wide geographical range covering 35 locations in 11 different regions from five countries. The intra-specific diversity of P. ariasi is high, with 45 haplotypes differing from each other by one to 26 bases and they are distributed in two mitochondrial lineages, one limited geographically to Algeria and the other widely dispersed across Mediterranean countries. The Algerian lineage is characterized by having 13 fixed polymorphisms and is made up of one sole haplotype. The European/Moroccan P. ariasi lineage is characterized by being made up of a great diversity of haplotypes (44) which display some geographical structuring. This could be one of the multiple factors involved in the epidemiological heterogeneity of the foci of leishmaniasis. Phlebotomus chadlii is the sister group of European/Moroccan P. ariasi. The separation of the Algerian haplotype, H45, from the rest of the specimens, European/Moroccan P. ariasi and P. chadlii, is well supported by the bootstrap analysis.     

Expression profiling of the triterpene saponin biosynthesis genes FPS, SS, SE, and DS in the medicinal plant Panax notoginseng

Panax notoginseng (Burk) F. H. Chen, an economically significant medicinal plant with hemostatic and health tonic activities, has been used in Traditional Chinese Medicine (TCM) for more than 3000 years. Triterpene saponins are responsible for most of the pharmacological activities of P. notoginseng. Here, we cloned five cDNA sequences encoding the key enzymes involved in triterpene saponin biosynthesis, namely, PnFPS, PnSS, PnSE1, PnSE2, and PnDS, and analyzed the conserved domains and phylogenetics of their corresponding proteins. Their organ-specific expression patterns in four-year-old P. notoginseng were detected by real-time PCR, showing that they were all most highly expressed in flowers. In addition, four of the genes, excluding PnSE2, were upregulated in leaves following stimulation with methyl jasmonate. This study is the first comprehensive analysis of the expression patterns of pivotal genes for triterpene saponin biosynthesis in P. notoginseng and provides a basis to further elucidate the molecular mechanism for the biosynthesis of these medically important compounds.     

Xylem of early angiosperms: Novel microstructure in stem tracheids of Barclaya (Nymphaeaceae)

Pit membranes of stem tracheids of all recognized species of Barclaya, an Indomalaysian genus of Nymphaeaceae, were studied with scanning electron microscopy (SEM). Pit membranes of the tracheids are composed of two thick layers, both constructed of fibrils much larger than those of tracheary elements of angiosperms other than Nymphaeaceae. The outer (distal) layer, which comprises the continuous primary wall around the tracheids, is spongiform, perforated by porosities of relatively uniform size, and confined to or most prominent on end walls of stem tracheids. The second layer consists of thick widely spaced fibrils that are oriented axially and are laid down proximally (facing the cell lumen) to the first (outer) layer, although continuous with it. These axial fibrils are attached at their ends to the pit cavities. This peculiar microstructure is not known outside Nymphaeaceae except in Brasenia and Cabomba (Cabombaceae, Nymphaeales), and has not been previously described for Barclaya. The longitudinally oriented threads and strands in perforation plates of secondary xylem of wood and stems of a variety of primitive woody angiosperms (e.g., Illicium) are not homologous to the pit membrane structure observed in stem tracheids of Barclaya, which, like other Nymphaeaceae, has only primary xylem and no perforation plates. The tracheid microstructure reported here is different from pit structures observed in any other group of vascular plants, living or fossil. The tracheid stem microstructures of Barclaya and other Nymphaeaceae appear to be a synapomorphy of Nymphaeaceae and Cabombaceae, and need further study with respect to ultrastructure and function.     

First detailed description of Hispanomys bijugatus Mein and Freudenthal, 1971 (Rodentia, Cricetodontinae) from the Upper Aragonian of La Grive-Saint Alban (France): Biostratigraphical implications

The material of Hispanomys bijugatus (Rodentia, Cricetodontinae) from La Grive-Saint Alban (carrière Lechartier, fissure L3) is described for the first time and compared with all species of the genus known to date. As common in the Upper Aragonian populations of Hispanomys, this taxon evidences a low variability. H. bijugatus shows some progressive characters with respect to the remaining Aragonian congeneric species, such as the absence of labial and lingual cingula surrounding the upper and lower molar valleys respectively, the increase in the number of roots on the second lower molar, and the lost of mesolophs. This suggests that H. bijugatus, in spite of being one of the oldest species of the genus, is relatively derived with regard to the coeval congeneric species. Because H. bijugatus and H. decedens are believed to be closely related species within the same lineage, the fact that the former shows a more progressive dental morphology than the latter suggests that the unnamed fissure-fillings from La Grive and La Grive M (with H. decedens only) are older than La Grive L3 (with H. bijugatus only). The coexistence of both species at locality L5 suggests an intermediate age.     

Cloning and expression of boule and dazl in the Nile tilapia (Oreochromis niloticus)

The Deleted in Azoospermia (DAZ) family of RNA binding proteins consists of highly conserved genes boule, daz and daz-like (dazl) essential for germ cell development. boule is known for its unisexual meiotic expression in invertebrates and mammals, but meiotic-specific female expression plus meiosis-preferential male expression in trout, and meiosis-preferential bisexual expression in medaka. dazl shows highly conserved bisexual expression throughout gametogenesis in diverse species. Here we report the cloning and expression of boule and dazl in the Nile tilapia (Oreochromis niloticus), an important aquaculture fish. Molecular cloning and sequence analysis led to the identification of tilapia boule and dazl cDNAs. The predicted partial Boule contains a conserved RRM motif and Dazl has the C-terminal sequence. On a phylogenetic tree, tilapia Boule and Dazl are in separate clades of Boule and Dazl homologs from other species, indicating their divergence during early vertebrate evolution. By RT-PCR analysis, boule and dazl showed bisexual gonad-specific expression. By in situ hybridization analysis, both boule and dazl RNAs were restricted to female and male germ cells of adult gonads but absent in gonadal soma. In the ovary, boule and dazl RNAs were abundant in oocytes. In the testis, boule and dazl RNAs were prominent in meiotic spermatocytes but barely detectable in meiotic products. These data show that boule and dazl are expressed bisexually in germ cells and provide useful markers to study gametogenesis in the adult tilapia.     

Leishsmania (Leishmania) amazonensis infection: Muscular involvement in BALB/c and C3H.HeN mice

Recent studies have provided some insights into Leishsmania (Leishmania) amazonensis muscular infection in dogs, although, muscular disease due to leishmaniasis has been poorly documented. The aim of our study was to evaluate involvement of Leishmania in muscular infection of two distinct mouse strains (BALB/c and C3H.He), with different genetic backgrounds. BALB/c mice, susceptible to Leishmania infection, showed, at the beginning of infection, a great number of infected macrophages among muscle fibers; however, in C3H.He resistant mice, muscle fibers were less damaged than in BALB/c mice, but some parasitized macrophages could be seen among them. A follow up of the infection showed an intense inflammatory infiltrate mainly composed of infected macrophages in BALB/c muscles and the presence of amastigotes within muscle fibers; while C3H.He mice exhibited a moderate inflammatory infiltrate among skeletal muscle fibers and an absence of amastigotes. Total destruction of muscles was observed in BALB/c mice in the late phase of infection (day 90) while C3H.He mice showed a process of muscle repair. We concluded that: (1) the muscles of BALB/c mice were more affected by leishmaniasis than those of C3/H.He mice; (2) Leishmania amastigotes are capable of infecting muscular fibers, as observed in BALB/c mice; (3) as inflammatory infiltrate is less intense in C3H.He mice these animals are capable of restoring muscular fibers.