Molecular data confirm the validity of the Portuguese blenny (Parablennius ruber, Valenciennes, 1836) and its presence in Western Europe

DNA sequence analysis confirms the distinction between Parablennius ruber and Parablennius gattorugine , simultaneously validating the presence of the former species in Western Europe where it has been reported for >150 years. A possible scenario involving speciation of P. ruber at the Azores and subsequent transport of larvae to Europe, a process that may be still occurring nowadays, could explain this pattern of occurrence.     

Structural and ultrastructural analysis of embryonic development of Prochilodus lineatus (Valenciennes, 1836) (Characiforme; Prochilodontidae)

This survey was performed to characterize the embryogenesis of Prochilodus lineatus. Seven stages of embryo development were identified--zygote, cleavage, blastula, gastrula, segmentation, larval and hatching--after a period of incubation of 22 h (24 degrees C) or 14 h (28 degrees C). The following cleavage pattern was identified: the first plane was vertical (2 blastomeres); the second was vertical and perpendicular to the first (4 blastomeres); the third was vertical and parallel to the first (4 x 2); the fourth cleavage was vertical and parallel to the second (4 x 4); the fifth was vertical and parallel to the first (4 x 8); and the sixth cleavage was horizontal (64 blastomeres). At the blastula stage (3.0-4.0 h (24 degrees C); 1.66-2.0 h (28 degrees C)) irregular spaces were detected and periblast structuring was initiated. At the gastrula stage (4.0-8.0 h (24 degrees C); 3.0-6.0 h (28 degrees C)) the epiboly, convergence and cell movements, as well as the formation of embryonic layers, had begun. The segmentation stage (10.0-15.0 h (24 degrees C); 7.0-10.0 h (28 degrees C)) was characterized by a rudimentary formation of organs and systems (somites, optic vesicle and intestinal delimitation). The embryo at the larval stage (16.0-21.0 h (24 degrees C); 11.0-13.0 h (28 degrees C)) showed a free tail, more than 25 somites, an optic vesicle and a ready-to-hatch larval shape. The blastomeres at cleavage stage had disorganized nuclei indicating high mitotic activity. At gastrula, the blastomeres and the periblast had euchromatic nuclei and a large number of mitochondria and vesicles. The yolk was organized into globose sacs, which were dispersed into small pieces prior to absorption.     

Embryogenesis and early larval development in wild-caught Levantine scraper,Capoeta damascina (Valenciennes, 1842)

Levantine scraper, Capoeta damascina is a candidate species for future stock assessments, conservation studies, and hatchery efforts. Herein, we documented embryonic and early larval development, from egg activation to the exogenous feeding period, using morphological and histological landmarks. Embryos were obtained by in vitro fertilization from hormonally induced wild-caught broodstock, and subsequent development was monitored at temperatures coinciding with native conditions. Embryonic development from fertilization to hatch lasted ~105–110 hr. Larvae emerged with unpigmented eyes and body morphology, as well as an undifferentiated digestive tract. The mouth was closed at hatch by the oropharyngeal membrane and opened by the early endogenous feeding period. Trabeculae cartilage, quadrate bone, and Meckel's cartilage of the endoskeleton were present during the endogenous feeding period. During this period, the larvae underwent considerable changes in craniofacial morphology, locomotion, and organogenesis of the digestive tract. The cartilaginous floor of the neurocranium developed and the first four ceratobranchials appeared simultaneously at the end of endogenous feeding period. The digestive tract was differentiated into buccopharynx, esophagus, and small intestine during the endogenous feeding period. The intestinal valve and numerous longitudinal folds at the posterior region of the intestine formed together by the endo–exogenous feeding period. Major developmental events in retinogenesis occurred during the endogenous feeding period. When larvae entered exogenous feeding the mouth was fully-functional. Additionally, liver size and eye diameter increased. Our analysis of embryonic and early larval development in Levantine scraper aligned with other freshwater fishes.     

Structural and ultrastructural characteristics of the yolk syncytial layer in Prochilodus lineatus (Valenciennes, 1836) (Teleostei; Prochilodontidae)

The yolk syncytial layer (YSL) has been regarded as one of the main obstacles for a successful cryopreservation of fish embryos. The purpose of this study was to identify and characterize the YSL in Prochilodus lineatus, a fish species found in southeastern Brazil and considered a very important fishery resource. Embryos were obtained through artificial breeding by hormonal induction. After fertilization, the eggs were incubated in vertical incubators with a controlled temperature (28 degrees C). Embryos were collected in several periods of development up to hatching and then fixed with 2% glutaraldehyde and 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.3). Morphological analyses were carried out under either light, transmission or scanning electron microscopy. The formation of the YSL in P. lineatus embryos starts at the end of the cleavage stage (morula), mainly at the margin of the blastoderm, and develops along the embryo finally covering the entire yolk mass (late gastrula) and producing a distinct intermediate zone between the yolk and the endodermal cells. The YSL was characterized by the presence of microvilli on the contact region with the yolk endoderm. A cytoplasmic mass, full of mitochondria, vacuoles, ribosomes, endomembrane nets and euchromatic nuclei, indicated a high metabolic activity. This layer is shown as an interface between the yolk and the embryo cells that, besides sustaining and separating the yolk, acts as a structure that makes it available for the embryo. The structural analyses identified no possible barriers to cryoprotectant penetration.     

Alexander Böhm (1971–2012)

On November 28, 2012 Alexander (Alex) Böhm, a bacterial geneticist, died at age 41, only a few months after taking up a position as an assistant professor at the LOEWE Center for Synthetic Microbiology in Marburg, Germany. Earlier in 2012 Alex had been diagnosed with an aggressive form of thyroid cancer that left him little time to live his scientific and personal dreams.     

Photoresponse ontogeny and its relation to development of pineal organ and eye in larval bagrid catfish Mystus nemurus (Valenciennes)

The artificially reared bagrid catfish Mystus nemurus was observed for the histological development of the pineal organ and retina and photoresponse in a test tank at hatching to 14?d after hatching. The pineal organ was functional at hatching, and the lens-like tissue was partly ossified forming a pineal window at 6?d. The retina became morphologically functional when the outer segments of single cones were formed, and the eyes were innervated with the optic tectum at 18?h and rods were formed at 36?h. Long and thin single cones were not observed. The larvae exhibited undirected kinetic movement at hatching to 12?h and directed tactic swimming away from a torch after 18?h in response to a torch light. The photoresponse of the larvae was negative at hatching to 30?h and at 6?d to the end of the observation at 14?d, but neutral during a period at 36?h to 5?d. It was evident that the kinetic movement was mediated by light perception with the pineal organ, which was not capable of detecting directed signal information, and that the larvae were capable of directed tactic movement only when vision was involved. The vigorous negative phototaxis at 6–14?d was attributed to the improvement of photosensitivity of the retina and the pineal organ.     

Gene banking of the neotropical fish Leporinus obtusidens (Valenciennes, 1836): a protocol to freeze its sperm in the field

A practical sperm cryopreservation protocol using a dry-shipper and a diluent of simple composition is described for the neotropical fish Leporinus obtusidens (Valenciennes, 1836). The cooling rate of the dry-shipper and its period of useful time, established under laboratory conditions, were respectively 25.7-30.8 degrees C/min (between 0 and -60 degrees C) and 9 days after charging. Sperm donors were selected on the basis of their hyperemic genital papilla and the ability to ooze milt under gentle manual pressure, during the reproductive months of November to January. Milt volume (1.3+/-0.3 mL; n=9 fish), fresh sperm motility rate (93.3+/-2.5%; n=6 fish), and sperm concentration (10.9+/-3.0 x 10(9)spermatozoa/mL of milt) were obtained. The sperm cryopreservation experiments were conducted with the following cryoprotectants (all at 10%, before mixing with milt): dimethyl sulphoxide (DMSO; n=10 fish), methanol (n=6 fish), propanediol (n=6 fish) and ethylene glycol (n=5 fish). Glucose (5%) and hen's egg yolk (10%) made up the diluents containing DMSO, ethylene glycol or propanediol. Milk powder (10%) replaced hen's egg yolk in the diluent containing methanol. Distilled water (up to 100%) completed the diluent solutions. Milt freezing (in 0.5-mL straws) was performed in the dry-shipper after 1:5 (milt:diluent) dilution. Thawed sperm cryopreserved in DMSO-containing diluent and activated by 119 mM NaHCO(3) gave the highest motility rate (62+/-14%). The fertilizing capacity of L. obtusidens sperm was tested using the combination of DMSO-containing diluent as the cryoprotectant and 119 mM NaHCO(3) as the activating solution. Oocytes were obtained from artificial spawning and fertilized with different proportions of spermatozoa. The greatest rate of fertilization (74%) occurred when the ratio of about 112,000 motile spermatozoa:oocyte was used. Thus, a protocol to freeze L. obtusidens sperm can be elaborated as follows. Milt (<1.5 mL fish(-1)) was readily available only in November to January; a simple solution, composed of 10% DMSO (concentration before adding milt), 5% glucose, and 10% hen yolk egg, in distilled water, was used as sperm diluent; cooling rate of 25-30 degrees C/min, yielded in a portable dry-shipper, was adequate to freeze diluted milt (1:5; milt:diluent), in 5-mL straws; about 112,000 thawed motile spermatozoa:oocyte activated by 119 mM NaHCO(3) assured a fertilization rate of 74%.     

Tape blenny Neozoarces pulcher Steindachner, 1880 (Perciformes: Zoarcidae)—a sole valid species of the genus, its reproduction, and embryonal and larval development

On the basis of morphometric analysis of our collections of pairs of spawning fish of the genus Neozoarces, development of prolarvae of fish, and a survey of museum collections, a conclusion is made in respect to only one of the two species of the genus—tape blenny Neozoarces pulcher, Steindachner, 1880. Differences in morphometric characters and coloration of different fish are accounted for by a sharply expressed sexual dimorphism. The reproduction and the egg and prolarvae development in N. pulcher are described.     

Stratigraphic significance of siliceous microfossils collected during dives (off Peru, 5 °–6°S)

The geological evolution of the northern Peru convergent margin can be traced using samples collected during deep-sea dives of the submersible Nautile. In the Paita area (5°–6°S), the sedimentary sequence was intensively sampled along the main scarp of the middle slope area. It consists of Upper Miocene (7–9 Ma) to Pleistocene siltstone, sandstone and rare dolostone. The age distribution of these samples is the basis for a new geologic interpretation of the multichannel seismic line CDP3.Siliceous microfossils (both diatoms and radiolarians) show influence of both cold and temperature waters (local species mixed with upwelling ones). Diatom assemblages studied from the NP1-13 and NP1-15 dives bear a strong resemblance to assemblages from the Pisco Formation of southern Peru.Micropaleontological data from siliceous microfossils, provide evidence for two main unconformities, one is at the base of the Quaternary sequence and the other corresponds to a hiatus of 1 Myr, separating the Upper Miocene (7–8 Ma) sediments from uppermost Miocene (5–6 Ma) sediments.During the past 400 kyr, a wide rollover fold developed in the middle slope area associated with a major seaward dipping detachment fault. A catastrophic debris a valanche occurred as the results of an oversteepening of the landward flank of the rollover fold. The gravity failure of the slope, recognized by SeaBEAM and hydrosweep mapping, displaced enough material to produce a destructive tsunami which occurred 13.8 ± 2.7 kyr ago.     

Avian Plasmodium infection in field‐collected mosquitoes during 2012–2013 in Tarlac,Philippines

Global warming threatens to increase the spread and prevalence of mosquito‐transmitted diseases. Certain pathogens may be carried by migratory birds and transmitted to local mosquito populations. Mosquitoes were collected in the northern Philippines during bird migration seasons to detect avian malaria parasites as well as for the identification of potential vector species and the estimation of infections among local mosquito populations. We used the nested PCR to detect the avian malaria species. Culex vishnui (47.6%) was the most abundant species collected and Cx. tritaeniorhynchus (13.8%) was the second most abundant. Avian Plasmodium parasites were found in eight mosquito species, for which the infection rates were between 0.5% and 6.2%. The six Plasmodium genetic lineages found in this study included P. juxtanucleare ‐GALLUS02, Tacy7 (Donana04), CXBIT01, Plasmodium species LIN2 New Zealand, and two unclassified lineages. The potential mosquito vectors for avian Plasmodium parasites in the Philippines were Cq. crassipes, Cx. fuscocephala, Cx. quinquefasciatus, Cx. sitiens, Cx. vishnui, and Ma. Uniformis; two major genetic lineages, P. juxtanucleare and Tacy7, were identified.     

Spawning and larval development of Tegula euryomphala (Jones, 1844) (Trochoidea: Tegulidae) from La Herradura Bay,Chile

The spawning and larval development of Tegula euryomphala (Jones, 1844) were studied under laboratory conditions. The release of gametes was stimulated by raising temperature from 16 to 19?°C, and could be induced during several months of the year. Males always spawned first. Eggs were 155.6?μm in diameter and remained unattached to each other. Development was similar to that described for other Trochoidea and lasted 3–5?days at 19?°C. Larvae hatched approximately 24 h post-fertilization at the pre-torsional veliger stage. The larval protoconch of the metamorphic veliger measured 247?μm and was ornamented with six spiral threads and weakly ordered granulation. Larvae metamorphose without any natural substratum.     

Amphibious behaviour and feeding ecology of the four-eyed blenny (Dialommus fuscus, Labrisomidae) in the intertidal zone of the island of Santa Cruz (Galapagos, Ecuador)

The labrisomid Dialommus fuscus , endemic to the Galapagos archipelago, is a predator of crustaceans, which moves up and down the intertidal zone with the tides. Its divided eye and elongated body permit it to forage in the wet sections of the intertidal habitat during low tide, both on the rocks and below the water line and in tide pools. Compared with other intertidal fishes (e.g. Bathygobius sp.), it was present on the rock flat in significantly smaller tide pools nearer to the water line during low tide.     

Morphometrics,‘condition’and gonad development of the East African Sardinella gibbosa (Bleeker) and Sardinella albella (Valenciennes)*

Sardinella gibbosa and Sardinella albella of Dar es Salaam coast were studied from January 1969 to March 1970. Measurements on the standard length, weight, body depth, length and width of the gonads were taken and qualitative assessments of fat coverage on the alimentary canal and maturation stages were made. New size-groups of both the species appeared in the lamp-fishery at c. 6 cm SL between October 1969 and February 1970 and the largest specimens caught were 14.1 and 12.9 cm SL for the first and second species respectively. Length-weight equations of the two species show higher values of the regression coefficients for the females compared to the males. Fat content of fishes > 10 cm SL was high during March to May and low during August to December 1969. First maturation and spawning are probably attained at c. 10.5 and 10 cm SL in the first and second species respectively. Mature S. gibbosa occurred between August and November 1969 and probably the species has a single, annual spawning season of 3.4 months duration. Mature S. albella, 10.1–12.5 cm SL occurred between August 1969 and February 1970; mature females, 12.6–12.9 cm SL were encountered throughout the sampling period.