The development of the tooth pattern and dentigerous bones in Polypterus senegalus (Cladistia, Actinopterygii).

The formation sequence of the tooth-bearing bones and the tooth pattern in early ontogeny of Polypterus senegalus is investigated using transparent preparation, histological sections, and SEM. During the attachment step of the yolk-sac larva the first dermal bones and teeth are formed. Teeth appear simultaneously in the areas of the maxillary, dentary, dermopalatine, prearticular, and coronoid 1 along with the first separate anlagen of these bones. A monostichous arrangement of primary teeth is established on the maxillary, dentary, and dermopalatine. Polystichous tooth arrangements do not occur before the early pterolarval phase, and then only in connection with bones of the palate and inner dental arcades. Especially pronounced is the influence of tooth formation on the structure of the parasphenoid that becomes much thickened by accretion of denticulate platelets, but we found neither evidence for a distinct vomeral contribution to the parasphenoid, nor a composite origin of the ectopterygoid in ontogeny. First replacement teeth are found in association with the maxillary and dentary as early as the late apterolarval phase. Primary teeth are of a single general type, whereas from the pterolarval phase onward three tooth types can be distinguished that are restricted to certain tooth bearing bones. Relatively late in ontogeny, dermo-metapterygoid and entopterygoid become formed and colonised by teeth, whereas first branchial teeth and tooth plates appear earlier during the first phase of extrinsic larval feeding. Characteristics of development of the dentition are discussed in comparison with character states of other better known fossil and recent taxa among Actinopterygii and Sarcopterygii. Compared to the assumed basic pattern of actinopterygian fishes, Polypteriformes show a derived condition with respect to structure, arrangement, replacement, and differentiation of teeth, which arises in sequence during larval development. This also corresponds to observed changes of feeding behaviour and functional demands during larval life.     

The first record of Macrogyrodactylus species (Monogenea, Gyrodactylidae) on freshwater fishes in Senegal with the description of Macrogyrodactylus simentiensis sp. nov., a parasite of Polypterus senegalus Cuvier

The first record of monogenean parasites of the genus Macrogyrodactylus Malmberg, 1957 on freshwater fish in Senegal is presented. Macrogyrodactylus congolensis Prudhoe, 1957 from the skin and Macrogyrodactylus heterobranchii N’Douba et Lambert, 1999 from the gills of Clarias anguillaris L. were found, representing new host records for these parasites. On Polypterus senegalus Cuvier, three Macrogyrodactylus species were identified, Macrogyrodactylus polypteri Malmberg, 1957, Macrogyrodactylus simentiensis sp. nov. and Macrogyrodactylus sp. M. simentiensis sp. nov. can be readily distinguished from the other Macrogyrodactylus species by the size of its hamuli and the shape of its marginal hook sickles. The marginal hooks on the anterolateral lobes of M. simentiensis differ in size and shape from those on the posterior margin of the haptor. Measurements and drawings of the haptoral sclerites of all five identified species are provided.     

The lamina cribrosa of Ornithorhynchus (Monotremata, Mammalia)

Summary A vestigial and transitory lamina cribrosa was found in nestling platypus (Ornithorhynchus anatinus). The heads of two nest-young (180 and 333 mm length), one subadult and one adult Ornithorhynchus were serially sectioned and studied with special reference to the development of the nasal region. In nest-young Ornithorhynchus an irregularly shaped bar of cartilage develops at the foramen olfactorium advehens. In the subadult it is largely resorbed, and in the osseous skull of the adult it is completely lacking. Ontogeny and topographical relationships of this bar of cartilage indicate that it is part of a lamina cribrosa. It embraces the ramus medialis of the nervus ethmoidalis and the arteria ethmoidalis, as do the corresponding parts of the lamina cribrosa of Tachyglossus. Compared to other parts of the chondrocranium this bar develops late in ontogeny, as does the lamina cribrosa of other mammals. Therefore, it can be concluded that part of the lamina cribrosa is present for a short period during the ontogeny of Ornithorhynchus, contrary to earlier reports. As in many other water-adapted mammals, the olfactory system of Ornithorhynchus is reduced. This suggests that the rest of the lamina cribrosa of Ornithorhynchus is secondarily reduced. The common ancestor of Ornithorhynchus and Tachyglossidae most probably possessed a lamina cribrosa which can be traced back to the common mammalian stock. The lamina cribrosa developed only once in the phylogeny of mammals. Its lack in the adult Ornithorhynchus is not a reptilian character.     

Evolutionary trajectories of tooth histology patterns in modern sharks (Chondrichthyes,Elasmobranchii)

During their evolutionary history, modern sharks developed different tooth mineralization patterns that resulted in very distinct histological patterns of the tooth crown (histotypes). To date, three different tooth histotypes have been distinguished: (i) orthodont teeth, which have a central hollow pulp cavity in the crown, encapsulated by a prominent layer of dentine (orthodentine); (ii) pseudoosteodont teeth, which have their pulp cavities secondarily replaced by a dentinal core of porous dentine (osteodentine), encased by orthodentine; and (iii) osteodont teeth, which lack orthodentine and the whole tooth crown of which consists of osteodentine. The aim of the present study was to trace evolutionary trends of tooth mineralization patterns in modern sharks and to find evidence for the presence of phylogenetic or functional signals. High resolution micro-computed tomography images were generated for the teeth of members of all nine extant shark orders and the putative stem group †Synechodontiformes, represented here by three taxa, to examine the tooth histology non-destructively. Pseudoosteodonty is the predominant state among modern sharks and represents unambiguously the plesiomorphic condition. Orthodonty evolved several times independently in modern sharks, while the osteodont tooth histotype is only developed in lamniform sharks. The two shark orders Heterodontiformes and Pristiophoriformes showed highly modified tooth histologies, with Pristiophorus exhibiting a histology only known from batomorphs (i.e. rays and skates), and Heterodontus showing a histological difference between anterior and posterior teeth, indicating a link between its tooth morphology, histology and durophagous lifestyle. The tooth histotype concept has proven to be a useful tool to reflect links between histology, function and its taxonomic value for distinct taxa; however, a high degree of variation, especially in the pseudoosteodont tooth histotype, demonstrates that the current histotype concept is too simplistic to fully resolve these relationships. The vascularization pattern of the dentine might offer new future research pathways for better understanding functional and phylogenetic signals in the tooth histology of modern sharks.     

Opecoelids (Platyhelminthes, Digenea) from the fork-tailed threadfin bream Nemipterus furcosus (Valenciennes, 1830) (Perciformes, Nemipteridae), with preliminary keys to the problematic genera Macvicaria Gibson et Bray, 1982 and Neolebouria Gibson, 1976

The opecoelid species Macvicaria jagannathi (Gupta et Singh, 1985) Bijukumar, 1997 (new syn. Plagioporus deeghaensis Gupta et Gupta, 1988) and Neolebouria lineatus Aken’Ova et Cribb, 2001 are redescribed from Nemipterus furcosus, from the waters off New Caledonia. Provisional keys to the genera Macvicaria Gibson et Bray, 1982 and Neolebouria Gibson, 1976 are presented. The following new combinations are made: Macvicaria yamagutii (Gupta et Ahmad, 1977), M. puriensis (Gupta et Govind, 1984) and M. chilkai (Gupta et Govind, 1984).     

The development of the diencephalon in Xenopus

Summary The development of the diencephalon and the time of origin of neurons of thalamic nuclei were determined in Xenopus with 3H-thymidine autoradiography Isotope was administered into embryos, tadpoles and metamorphic animals and sacrificed after survival time between 24 hours and 5 months. The position and the number of heavily labeled cells, corresponding to terminal mitoses at the time of isotope injection were established on transverse and sagittal sections of the frog brain. Neurons in the diencephalon were distributed in a spatiotemporal manner such, that cells generated earliest were located in the caudo-ventro-lateral portion of the diencephalon followed in a rostro-dorso-medial sequence by cells formed at later stages. The waves of cell generation resulted in three apparent developmental gradients in the caudo-rostral, latero-medial and in the ventro-dorsal directions in the diencephalon. Consequently neurons generated latest were found in the rostro-dorso-medial portion of the diencephalon. the overall rostro-dorso-medial diencephalic growth and the spatiotemporal generation of its neurons are the reverse of the tectal growth and cell generation reported in Xenopus which occurs in a rostrolateral to caudomedial direction.The findings of the present observations appear to indicate that the mirror-image reversal of the retinotectal and retinodiencephalic projections along the temporo-nasal retinal axis is the consequence of the divergent growth of the diencephalon and the tectum from the common embryonic di-mesencephalic junction. These observations furthermore suggest that the orientation of the retinal maps is ensured by the differential maturity gradients in the tectum and diencephalon, respectively, presumably expressed in molecular terms.     

Identification of Rhodococcus, Gordona and Dietzia species using carbon source utilization tests (“Biotype-100” strips)

The “Biotype-100” identification system (BioMérieux, La Balme-les-Grottes, France) based on carbon source utilization was evaluated for its ability to discriminate among 10 species of Rhodococcus, 7 species of Gordona and one species of Dietzia. The type strains of three species of Tsukamurella and 8 species of Nocardia were also included in the study. Results were compared with chemotaxonomic and conventional data. Carbon source utilization was shown to be reliable, rapid and easy to use when compared with standard identification methods. The 29 species tested were unambiguously separated by carbon source utilization tests. Rhodococcus equi was found to be heterogenous.Les galeries “Biotype-100” (BioMérieux) ont été utilisées pour différencier 10 espèces du genre Rhodococcus, 7 espèces du genre Gordona et 1 espèce du genre Dietzia entre elles. Les souchestypes de 3 espèces du genre Tsukamurella et 8 espèces du genre Nocardia ont été incluses dans cette étude. Les résultats ont été comparés avec ceux des études chimiotaxonomiques et ceux obtenus avec les galeries d'identification classiques. Les galeries Biotype-100 sont sûres, rapides et faciles à utiliser par rapport aux galeries classiques. Les 29 espèces étudiées ont été identifiées sans aucune difficulté. L'espèce Rhodococcus equi s'est révélée hétérogène.     

Synergistic interaction between the Potyvirus, Turnip mosaic virus and the Crinivirus, Lettuce infectious yellows virus in plants and protoplasts

Lettuce infectious yellows virus (LIYV), the type member of the genus Crinivirus in the family Closteroviridae, is specifically transmitted by the sweet potato whitefly (Bemisia tabaci) in a semipersistent manner. LIYV infections result in a low virus titer in plants and protoplasts, impeding reverse genetic efforts to analyze LIYV gene/protein functions. We found that synergistic interactions occurred in mixed infections of LIYV and Turnip mosaic virus (TuMV) in Nicotiana benthamiana plants, and these resulted in enhanced accumulation of LIYV. Furthermore, we examined the ability of transgenic plants and protoplasts expressing only the TuMV P1/HC-Pro sequence to enhance the accumulation of LIYV. LIYV RNA and protein titers increased by as much as 8-fold in these plants and protoplasts relative to control plants. LIYV infections remained phloem-limited in P1/HC-Pro transgenic plants, suggesting that enhanced accumulation of LIYV in these plants was due primarily to increased replication efficiency, not to greater spread.     

Antifungal activity of Pterocaulon species (Asteraceae) against Sporothrix schenckii

Plants of the genus Pterocaulon (Asteraceae) are popularly used in the treatment of skin diseases caused by fungi and bacteria. The aim of this work was to investigate the in vitro activity of the crude methanolic extracts obtained from the aerial parts of Pterocaulon polystachyum, P. balansae, P. lorentzii, P. lanatum, and P. cordobense against 24 Sporothrix schenckii clinical isolates and determine the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC). MIC were performed by the broth microdilution method according guidelines recommended by Clinical and Laboratory Standards Institute for filamentous fungi and MFC were determined for transference of aliquots of the well that showed 100% of growth inhibition into tubes with culture medium. The extract from P. polystachyum was the most active sample, presenting MIC range of 156 and 312 μg/mL. The popular use of these plants corroborates the importance of ethnopharmacological surveys and opens the possibility for finding new clinically effective antifungal agents.     

RNAi knock-down of the Litopenaeus vannamei Toll gene (LvToll) significantly increases mortality and reduces bacterial clearance after challenge with Vibrio harveyi

In this study, we used real-time PCR to simultaneously monitor the responses of 12 key genes of the shrimp innate immune system in Litopenaeus vannamei after challenge with Vibrio harveyi. In the proPO activating system, we found that proPO was up-regulated (3.3× control at 36 hpi). The hemolymph clotting genes transglutaminase (TGase) and clotting protein were also up-regulated, as were 5 genes in the antimicrobial peptide system (ALF, Crustin, Lyz, PEN2 and PEN4), with only PEN3 showing no significant changes. In the antioxidant defense system, SOD was slightly elevated while GPx was substantially down-regulated. In the pattern recognition receptor system, at 24 hpi, the Toll gene (LvToll) showed the highest relative increase in expression level of all the investigated genes (15× greater than the sterile seawater control). In the second part of this study, when LvToll was knocked down by RNAi silencing, there was no effect on either survival rates or bacterial number in unchallenged shrimp. There was also no difference in mortality rates between control shrimp and LvToll-silenced shrimp when these two groups were challenged with a viral pathogen (white spot syndrome virus; WSSV). However, when LvToll-silenced shrimp were challenged by V. harveyi, there was a significant increase in mortality and bacterial CFU counts. We note that the increase in bacterial CFU count occurred even though treatment with EGFP dsRNA had the opposite effect of reducing the CFU counts. We conclude that LvToll is an important factor in the shrimp innate immune response to acute V. harveyi infection, but not to WSSV.     

Some gonad-infecting species of Philometra (Nematoda, Philometridae) from offshore fishes of South Carolina and Georgia, USA, including Philometra charlestonensis sp. nov. from the scamp Mycteroperca phenax

Three gonad-infecting species of Philometra Costa, 1845 were, for the first time, recorded from perciform fishes from estuarine and marine waters in South Carolina and Georgia, USA: Philometra charlestonensis sp. nov. from the scamp Mycteroperca phenax (Jordan et Swain) (Serranidae), P. saltatrix Ramachandran, 1973 from the bluefish Pomatomus saltatrix (Linnaeus) (Pomatomidae), and Philometra sp. from the Atlantic croaker Micropogonias undulatus (Linnaeus) (Sciaenidae). The new species is characterized mainly by males (body length 2.65–3.14 mm) with equally long, needle-like spicules (length 132–141 μm) and the gubernaculum (81–93 μm) bearing dorsal transverse lamella-like structures on its distal portion, the body length of gravid females (168–247 mm), the presence of a well-developed anterior bulbous inflation on the female oesophagus, and by the length of the first-stage larvae (544–597 μm). A key to gonad-infecting species of Philometra parasitizing marine and brackish-water fishes is provided.     

Sarcocystis arctosi sp. nov. (Apicomplexa, Sarcocystidae) from the brown bear ( Ursus arctos ), and its genetic similarity to schizonts of Sarcocystis canis -like parasite associated with fatal hepatitis in polar bears ( Ursus maritimus )

The tissues of herbivores are commonly infected with cysts of parasites belonging to the apicomplexan genus Sarcocystis, but such sarcocysts are rare in bears. Here, we describe a new species, Sarcocystis arctosi, based on the mature sarcocysts identified in two brown bears (Ursus arctos) from Alaska, USA. Microscopic sarcocysts (37–75 × 20–42 μm) had thin walls (<1 μm). The outer layer of the sarcocyst, the parasitophorous vacuolar membrane (pvm), was wavy in outline and had minute undulations that did not invaginate towards the sarcocyst interior; these undulations occurred at irregular intervals and measured up to 100 nm in length and up to 60 nm width. The ground substance layer beneath the pvm was smooth and lacked microtubules. Longitudinally cut bradyzoites measured 5.6–6.8 × 0.7–1.8 μm. A major portion of nuclear small subunit rDNA sequence obtained from these sarcocysts was similar to that previously obtained from the hepatic schizonts of a S. canis-like parasite from polar bears (Ursus maritimus).     

A new species of Stephanostomum Looss, 1899 (Digenea, Acanthocolpidae) with a bizarre oral sucker: S. adlardi sp. nov. from the common coral trout Plectropomus leopardus (Lacepède, 1802) (Perciformes, Serranidae) from Lizard Island, Great Barrier Reef

A new species of Acanthocolpidae, Stephanostomum adlardi is described from the serranid Plectropomus leopardus from Lizard Island in the northern Great Barrier Reef. It differs from all previously described acanthocolpids in the structure of the oral sucker which is extended into dorsal and ventral lobes each bearing a row of spines. A phylogenetic tree estimated from combined nuclear small and partial large ribosomal RNA gene sequences shows that, despite the unusual oral sucker structure, the species is a true member of the genus Stephanostomum. The molecular results also suggest that Monostephanostomum nolani is derived from within Stephanostomum.     

Isolation and comparison of the IgM heavy chain constant regions from Australian (Trichosurus vulpecula) and American (Monodelphis domestica) marsupials

cDNAs encoding IgM heavy chain constant region (Cμ) were isolated from two metatherians (marsupials) — the Australian common brushtail possum (Trichosurus vulpecula) and the South American grey short-tailed opossum (Monodelphis domestica). Analysis of the sequences suggested that they correspond to the secreted form of Cμ in both species. The domain size and structure of the marsupial Cμ sequences were compared with other Cμ sequences and a high degree of conservation throughout vertebrate evolution was observed. Amino acid sequence comparisons revealed a marked level of sequence similarity between the two marsupial sequences (79%), relatively high similarity between the marsupials and eutherians (63%), and lower similarities between marsupials and birds (45%), marsupials and amphibians (47%), marsupials and reptiles (45%) and marsupials and fish (37%). These data allow the incorporation of metatherians into the study of mammalian IgM evolution.     

Loss of λ prophage recombinogenicity in UV-irradiated Escherichia coli: the role of host genes ruvA, ruvB, ruvC, and recG

Earlier studies have revealed a radiation-induced process leading to the loss of λ prophage recombinogenicity. The process takes place in UV-irradiated Escherichia coli cells, and renders the prophage incapable of site-specific recombination with the host chromosome, and of general recombination with an infecting homologous phage. It was found that the inhibition of prophage recombinogenicity depends on functional RecBCD enzyme of E. coli. In this work, the role of ruvABC and recG genes in the inhibitory process was assessed. The products of these genes are known to act at the last step of homologous recombination and recombinational DNA repair by catalyzing the resolution of recombination intermediates (the Holliday junctions). Irradiated prophage retained its ability to recombine in ruvA, ruvB, ruvC, and recG mutants. These results suggest that in addition to RecBCD enzyme, RuvABC and RecG proteins are also involved in the inhibition of prophage recombinogenicity. We infer that RuvABC and RecG act in this process before RecBCD, probably by processing the Holliday junctions formed upon replication arrest, and thereby providing double-stranded DNA breaks as substrate for RecBCD-mediated recombinational repair of UV-damaged bacterial chromosome.     

The heme-binding protein (HbpA) of Haemophilus influenzae as a virulence determinant

Haemophilus influenzae has an absolute growth requirement for heme and the heme-binding lipoprotein (HbpA) and has been implicated in the utilization of this essential nutrient. We constructed an insertional mutation of hbpA in a type b and a nontypeable H. influenzae strain. In the type b strain, the hbpA mutant was impaired in utilization of heme complexed to either hemopexin or to albumin and in the utilization of low levels of heme but not in the utilization of heme at high levels or of hemoglobin or hemoglobin–haptoglobin complexes. In contrast, the hbpA mutant derivative of the nontypeable strain was impaired in utilization of all tested heme sources. We further examined the impact of the hbpA mutation in animal models of H. influenzae disease. The hbpA mutant of the nontypeable strain was indistinguishable from the wild-type strain in the chinchilla model of otitis media. The hbpA mutant derivative of the type b strain caused bacteremia as well as the wild-type strain in 5-day old infant rats. However, in 30-day old rats the hbpA caused significantly lower rates of bacteremia than the wild-type strain indicating a role for hbpA and heme acquisition in virulence in this model of H. influenzae disease. In conclusion, HbpA is important for heme utilization by multiple H. influenzae strains and is a virulence determinant in a model of H. influenzae invasive disease.     

Molecular and functional characterization of kita and kitla of the goldfish (Carassius auratus L.)

Kit ligand and its type III tyrosine kinase receptor Kit promotes the survival, proliferation and differentiation of progenitor cells involved in mammalian myelopoiesis. In this study we report on the molecular and functional characterization of kit receptor A (kita) and kit ligand A (kitla) from the goldfish. Both kita and kitla were ubiquitously expressed in goldfish tissues, with higher mRNA levels observed in the kidney and spleen, the major hematopoietic organs of fish. Furthermore, both kita and kitla expressions decreased in a time-dependent manner in goldfish primary kidney macrophage (PKM) cultures, as progenitor to macrophage development progressed, and the highest expressions of both the receptor and ligand were observed in sorted progenitor cell populations. Activation of mature macrophage cultures increased both kita and kitla expressions. Kit ligand A induced chemotactic response, proliferation and survival of PKM cells in a dose-dependent manner, but did not induce differentiation of early PKM cells. These results are consistent with the role of kita and kitla during myelopoiesis of higher vertebrates and suggest a conserved mechanism of macrophage development throughout vertebrates.     

Ontogeny of the endocrine cells of the intestine and rectum of sea bass (Dicentrarchus labrax L.): an ultrastructural study

Several endocrine cell types were ultrastructurally characterized during the differentiation of the intestine and rectum of sea bass (Dicentrarchus labrax L.) larvae. Only one cell type (type I) was found in the posterior region of the undifferentiated gut of 5-day-old larvae (phase I). Types V and VI were found in both the intestine and rectum, types II, III and IV in the intestine, and types VII and VIII in the rectum of 9- and 12-day-old larvae (phase II), the rectum alone showing signs of functional differentiation. In phase III larvae, in which both the intestine and rectum were differentiated, types IX, X, XI, XII, XIII, XIV and XV were found in the intestine, only types X, XI and XII being seen in the rectum. Besides these, a new cell type, XVI, was observed in the intestine of 55- and 60-day-old larvae (phase IV), in which the digestive tract was completely differentiated. The endocrine cells appearing in phases I and II showed very scarce secretory granules and the ultrastructural features of undifferentiated cells. Some endocrine cell types in the earliest developmental stages were related to some of those found later. A maturational process of the endocrine cell types paralleled the differentiation of the intestine and rectum, with an apparent increase in the number of secretory granules accompanying organelle development.