Inception of bioluminescent symbiosis in early developmental stages of the deep-sea fish,Coelorinchus kishinouyei (Gadiformes: Macrouridae)

Larvae and juveniles of the macrourid fish Coelorinchus kishinouyei, captured from the near-bottom habitat (ca. 1–10 m above the seafloor) at 186 to 500 m depth in Suruga Bay, Honshu, Japan, were examined for the presence, developmental state, and bacterial colonization of the fish’s internal ventral light organ. The specimens ranged from 3.6 mm to 8.5 mm head length, and all exhibited an external cluster of melanophores expanding anteriorly from around the anus that is thought to indicate the presence of an internal light organ. Histological analysis revealed the presence of a light organ in all examined specimens. In smaller specimens, the light organ was seen as a small nub of tissue associated with the intestine near the anus; the light organ gradually elongated anteriorly in larger specimens to form a bean-shaped structure composed of hollow, finger-like chambers. Bacteria were present within the light organ chambers of some, but not all larvae and all juveniles. In light organs not yet colonized by bacteria, the chambers exhibited a generally uniform appearance over their entire length. In colonized light organs, the bacteria were consistently present at the anterior-most tips of the chambers; furthermore, cells comprising chambers colonized by bacteria were swollen, and upon bacterial colonization the orientation of the chambers began to change from anterior–posterior to dorsal–ventral. The colonizing bacteria were identified as Photobacterium kishitanii based on sequence analysis of the luxA gene. These results suggest that formation of the light organ in C. kishinouyei begins during the fish’s pelagic phase, but that bacterial colonization of the light organ occurs after the larvae have reached the near-bottom habitat. Furthermore, colonization of the nascent light organ by P. kishitanii induces morphogenetic changes in the light organ.     

Osmotic control of luminescence and growth in Photobacterium leiognathi from ponyfish light organs

Osmolarity was found to control the luminescence and growth of Photobacterium leiognathi strain LN-1a isolated from the light organ of the ponyfish Leiognathus nuchalis (family Leiognathidae). Low osmolarity (ca. 400 mOsm) stimulated luminescence per cell 80 to 100-fold to a level (ca. 2.0×10⁴ quanta·s^-1·cell^-1) equal to that of bacteria taken directly from the light organ and increased the level of luciferase per cell 8 to 10-fold compared to high osmolarity (ca. 800 mOsm). Conversely, high osmolarity stimulated oxygen uptake and growth rate 2 to 4-fold compared to low osmolarity. Of 21 additional tested strains of P. leiognathi from light organs of 9 other ponyfish species, all responded similarly. Low osmolarity may be a host control factor that functions to stimulate the luminescence and restrict the growth of ponyfish light organ bacteria in situ.     

Development of Sebastes taczanowskii (Scorpaenidae) in the Sea of Japan off Hokkaido with a key to species of larvae

The larval and juvenile stages of Sebastes taczanowskii (Japanese name: Ezo-mebaru) are described and illustrated based on 33 wild specimens [7.1–26.9 mm in body length (BL)] collected in the Sea of Japan, and eight specimens of reared larvae extruded from the one specimen of a captive pregnant female. Larvae were extruded between 4.3–5.0 mm BL and notochord flexion occurred 5.7–9.0 mm BL. Transformation from postflexion larvae to pelagic juveniles occurred between 13 and 17 mm BL. Preflexion and flexion larvae have a single melanophore row on the dorsal surface on the tail, and an internal line of melanistic dashes on the ventral side of the tail. Lateral pigmentation of postflexion and transforming larval body surfaces are light. Compared with other Japanese rockfish species, S. taczanowskii is shallow-bodied throughout both larval and juvenile stages. We provide an identification key to preflexion and flexion stage rockfish larvae found around the Japanese archipelago, and comparisons with other species. Larval and juvenile S. taczanowskii occurred in both near-shore and relatively offshore water around Shakotan Peninsula-Ishikari Bay, Hokkaido in June and July.     

The specificity of symbiosis: Pony fish and luminescent bacteria

Luminescent bacteria isolated from light organs of seven different species (3 genera) of fishes of the family Leiognathidae were subjected to taxonomic analysis. Of the 733 isolated all but seven were identified as Photobacterium leiognathi; the others are considered to be either chance contaminants of the sampling procedure or transients within the organ. In most fish, the luminous organ appeared to contain a single predominating strain of P. leiognathi with small numbers of one to three other strains of the same species, differing by only one or two characters.     

First report on the spawning migration and early life development of a cyprinid species of the genus Telestes

Reproductive migration of a Telestes species is reported for the first time for the Balkan endemic Telestes pleurobipunctatus (Stephanidis, 1939) from Louros River (Northwestern Greece), while the embryonic and larval development is also described for the first time for this species and genus. Adult fish migrate in late winter upstream and spawn in a peak of 2–3 nights in the outflow of springs. Embryos at hatching measure approx. 6.0 mm notochord length (NL), flexion starts at 8.1 mm NL and food particles are noted in the digestive tract of larvae at 8.2 mm NL. The larval period is characterized by the sequential formation of fins and the development of scalation. Fin differentiation ends with the completion of pelvic fin rays and the disappearance of the pre-anal finfold at 20.0 mm. First scales appear at 17.0 mm standard length (SL) and scalation completes at 27.8 mm SL. We propose that the size at a specific ontogenetic event, the pigmentation pattern and the pre-anal myomeres' number comprise diagnostic characters that permit distinguishing T. pleurobipunctatus larvae from other sympatric cyprinids. We note the rare pattern of scale scalation and pigmentation pattern of the target species. Parapatric speciation in T. pleurobipunctatus possibly related to its spawning migration pattern, as well as the use of larval identification in monitoring and conservation programmes targeting threatened cyprinids exhibiting this type of reproductive behaviour are also discussed.     

Phylogenetic Diversity and Cosymbiosis in the Bioluminescent Symbioses of “Photobacterium mandapamensis”

“Photobacterium mandapamensis” (proposed name) and Photobacterium leiognathi are closely related, phenotypically similar marine bacteria that form bioluminescent symbioses with marine animals. Despite their similarity, however, these bacteria can be distinguished phylogenetically by sequence divergence of their luminescence genes, luxCDAB(F)E, by the presence (P. mandapamensis) or the absence (P. leiognathi) of luxF and, as shown here, by the sequence divergence of genes involved in the synthesis of riboflavin, ribBHA. To gain insight into the possibility that P. mandapamensis and P. leiognathi are ecologically distinct, we used these phylogenetic criteria to determine the incidence of P. mandapamensis as a bioluminescent symbiont of marine animals. Five fish species, Acropoma japonicum (Perciformes, Acropomatidae), Photopectoralis panayensis and Photopectoralis bindus (Perciformes, Leiognathidae), Siphamia versicolor (Perciformes, Apogonidae), and Gadella jordani (Gadiformes, Moridae), were found to harbor P. mandapamensis in their light organs. Specimens of A. japonicus, P. panayensis, and P. bindus harbored P. mandapamensis and P. leiognathi together as cosymbionts of the same light organ. Regardless of cosymbiosis, P. mandapamensis was the predominant symbiont of A. japonicum, and it was the apparently exclusive symbiont of S. versicolor and G. jordani. In contrast, P. leiognathi was found to be the predominant symbiont of P. panayensis and P. bindus, and it appears to be the exclusive symbiont of other leiognathid fishes and a loliginid squid. A phylogenetic test for cospeciation revealed no evidence of codivergence between P. mandapamensis and its host fishes, indicating that coevolution apparently is not the basis for this bacterium's host preferences. These results, which are the first report of bacterial cosymbiosis in fish light organs and the first demonstration that P. leiognathi is not the exclusive light organ symbiont of leiognathid fishes, demonstrate that the host species ranges of P. mandapamensis and P. leiognathi are substantially distinct. The host range difference underscores possible differences in the environmental distributions and physiologies of these two bacterial species.     

Allometric and ontogenetic larval development of common barbel during rearing under optimal conditions

The rearing of finfish larvae is a key element in their further culture. Improper breeding protocols may result in high mortality rates, body deformation and growth rate decreases in both the larval and fattening periods. These errors can be avoided by thorough exploration of various aspects of early larvae biology, at least in model fish species. In this study, anatomical and morphological developments were analysed using allometric growth patterns of common barbel, Barbus barbus, larvae reared under optimal controlled conditions. Larvae of common barbel, which is a model species for fish of the genus Barbus, were reared for 30 days at 25 °C in the recirculated aquaculture system (RAS). Four periods of the barbel larval development were identified: pre-flexion (0–5 days post hatching – DPH; total length – TL: 9.5 ± 0.3 to 12.3 ± 0.3 mm), flexion (6–11 DPH; TL 12.4 ± 0.3–15.4 ± 0.3 mm), post-flexion (12–21 DPH; TL 16.1 ± 0.5–21.2 ± 0.8 mm) and juvenile (from 22 DPH; TL from 21.4 ± 1.7 mm). The largest changes in barbel growth were observed during the first two periods of their life (pre-flexion and flexion), which resulted in the frequency of noted flexion points (64.3% flexion points) and was also associated with intensive morphometric growth, primarily the head and tail parts of the body. Despite a low degree of growth progress upon hatching (e.g. no eye pigment, no distinct liver or pancreas, no unobstructed alimentary tract), barbel larvae pass through the larval periods very quickly in comparison to other cyprinids and enter the juvenile period (22 days).     

Growth and morphological development of laboratory-reared larval and juvenile three-spot gourami <Emphasis Type="Italic">Trichogaster trichopterus</Emphasis>

Morphological development, including the body proportions, fins, pigmentation and labyrinth organ, in laboratory-hatched larval and juvenile three-spot gourami Trichogaster trichopterus was described. In addition, some wild larval and juvenile specimens were observed for comparison. Body lengths of larvae and juveniles were 2.5 ± 0.1 mm just after hatching (day 0) and 9.2 ± 1.4 mm on day 22, reaching 20.4 ± 5.0 mm on day 40. Aggregate fin ray numbers attained their full complements in juveniles >11.9 mm BL. Preflexion larvae started feeding on day 3 following upper and lower jaw formation, the yolk being completely absorbed by day 11. Subsequently, oblong conical teeth appeared in postflexion larvae >6.4 mm BL (day 13). Melanophores on the body increased with growth, and a large spot started forming at the caudal margin of the body in flexion postlarvae >6.7 mm BL, followed by a second large spot positioned posteriorly on the midline in postflexion larvae >8.6 mm BL. The labyrinth organ differentiated in postflexion larvae >7.9 mm BL (day 19). For eye diameter and the first soft fin ray of pelvic fin length, the proportions in laboratory-reared specimens were smaller than those in wild specimens in 18.5–24.5 mm BL. The pigmentation pattern of laboratory-reared fish did not distinctively differ from that in the wild ones. Comparisons with larval and juvenile morphology of a congener T. pectoralis revealed several distinct differences, particularly in the numbers of myomeres, pigmentations and the proportional length of the first soft fin ray of the pelvic fin.     

Arms as areas for larval development of migratory fish species in a Neotropical reservoir and the influence of rainfall over abundances

The aim of the present study was to verify the use of the arms of the Itaipu Reservoir as areas of initial development for migratory fish species and to assess the relationship between rainfall and the spawning of migratory fish. Accordingly, fish larvae were collected from five arms of the reservoir from 2009 to 2016 using 0.5 mm plankton nets. Density was standardized as the number of larvae per 10 m³ filtered water, and the captured larval and juvenile specimens were identified at the lowest-possible taxonomic level. The larvae were also classified according to the degree of development and notochord flexion stage: larval vitelline, pre-flexion, flexion and post-flexion. To evaluate the distribution of larval abundance and the developmental stage along the longitudinal gradients of the arms, the data were evaluated using a set of nested linear models, following the AIC and Bayesian information criteria. In addition, an analysis of covariance was performed to investigate the influence of rainfall on the larval abundance of migratory species. During sampling, several species of economic and conservation interest such as Salminus brasiliensis and Pseudoplatystoma corruscans were collected. The larvae of the migratory fish taxa were captured from all sampled arms, which indicate them as areas of initial development. Nevertheless, it was observed that larval density increases from fluvial towards lacustrine zones inside the arms. Also, the present study verified that species, even in lentic environments, respond positively to rainfall stimuli in a manner similar to that exhibited by conspecifics in lotic environments. Such results reinforce the necessity of the protection of arms aiming at the conservation of this main group of species impaired by the construction of dams.     

Ontogenic changes in tolerance to hypoxia and energy metabolism of larval and juvenile Japanese flounder Paralichthys olivaceus

Changes in tolerances to hypoxia and sodium azide, an indicator of cellular respiration, and activities of various energy metabolism-related chemical components were studied in Japanese flounder Paralichthys olivaceus during its early life stages from 3.5 to 20.5 mm in total length (TL). They showed flexion stage around 10.4 mm TL. Lethal levels of hypoxia increased with growth from 3.5 to 8 mm total TL, and the levels remained high in larvae, until 10.4 mm TL, decreased significantly thereafter. The 50% lethal concentration of sodium azide temporarily increased at 4.5 mm TL, diminished drastically between 4.5 and 10.4 mm TL, and then increased again in post-flexion larvae. Cytochrome c oxidase activity was highest in larvae around flexion, at 10.4 mm TL, and subsequently decreased. In post-flexion larvae at 13.0 mm TL, lactate dehydrogenase (LDH) and creatine kinase activities increased; LDH activity decreased at the juvenile stage. The adenosine triphosphate content and energy charge in fish were consistently higher in the larval stage than in the juvenile stage. These results indicated that, from just before flexion to the post-flexion stage, the energy metabolism of larvae is higher due to activated aerobic and subsequent anaerobic metabolism for metamorphosis; as a consequence, hypoxia tolerance in fish is the lowest during the increase of aerobic metabolism just before and around flexion.     

Seasonal and Geographic Distribution of Luminous Bacteria in the Eastern Mediterranean Sea and the Gulf of Elat

Luminous bacteria in the Mediterranean Sea and the Gulf of Aqaba-Elat have different distribution patterns. In the Mediterranean Sea, Beneckea harveyi is present all year round, with different subtypes alternating in summer and winter; Photobacterium fischeri was only present during the winter. In the Gulf of Elat, P. leiognathi is present throughout the water column in similar densities during the entire year. This constancy in distribution is presumably due to the near-constancy in water temperature. In summer, Photobacterium leiognathi is replaced by B. harveyi in coastal surface waters. In the hypersaline Bardawil lagoon, only B. harveyi types are present. P. fischeri, a major component of the Mediterranean Sea winter communities, is absent from the lagoon. Luminous Beneckea strains show a great diversity in properties, e.g. temperature range for growth, sensitivity to infection by phages, sensitivity to attack by Bdellovibrio strains, and differences in tolerance to high-salinity shock. Therefore, subdivision of the taxonomic cluster of B. harveyi into subtypes is indicated. The composition of the luminous bacteria communities may serve as indicators of different marine water bodies. The symbiotic luminous bacteria of the light organ of the common Gulf of Elat fish, Photoblepharon palbebratus steinitzi, is different from any of the types described.     

The Luminescent Systems of Pony Fishes

The anatomy of bioluminescent organs and mode of light production in 18 species of pony fish have been investigated using fresh and preserved material. The luminescent systems are similarly arranged in all. Basically, the system consists of a light organ located at the distal end of the esophagus, and a series of abdominal accessory structures positioned in tandem for controlling light intensity and for directing and dispersing the light. Light is produced by numerous symbiotic luminous bacteria in the light organ. A simple classification of the luminescent systems is proposed. The light organs of Leiognathus elongatus and L. rivulatus show marked sexual dimorphism. The bacteria present in the light organs of many pony fishes are easily culturable, but not those from L. elongatus. Electron micrographs of the light organs of L. elongatus and L. rivulatus show the presence of numerous rod-shaped bacteria measuring approximately 0.8 µ x 2.4 µ and 0.8 µ x 7.3 µ, respectively. It is concluded that the light organ of L. elongatus contains another example of a type of non-culturable luminous bacteria that have been found elsewhere. Such bacteria appear to require from the host some special factor for growth and luminescence.     

Osteological development of the lophiid anglerfish,lophius gastrophysus

The osteological development of the head skeleton and dorsal, pectoral, and anal fin supports, are described from cleared and stained specimens ofLophius gastrophysus larvae, ranging from 4.6 to 21.8 mm NL; the results are compared with those of juvenile (79.8 mm SL) and adult (398 mm SL) specimens. Tiny conical teeth are present on the premaxillary, dentary, palatine and vomer since early stage. The first three dorsal fin spines are initially positioned on the midline of body posterior to the supraoccipital, but they migrate forward with growth and become cephalic in juveniles. The forward movement of the dorsal spines is produced by the forward extension of the cartilaginous basal inside the subepidermal space. During the planktonic larval stage the pectoral fins are on the sides of body as in ordinary fishes, but they move ventrad and become leg-like in bottom living juveniles and adults. Ossification of the caudal complex ofL. gastrophysus larvae proceeds very slowly and only the 21.8 mm NL larva has an almost completely ossified caudal complex. Eight principal caudal rays are loosely attached on the posterior edge of the hypurals and no procurrent rays are present. Larvae have well developed parhypurapophysis at the mid-portion of the urostyle which transforms into keel-like structure in juveniles and adults.     

Morphological aspects of the development of swimming and feeding functions in the milkfishChanos chanos

Development of swimming and feeding abilities based on morphological development of larval and early juvenileChanos chanos was investigated. In larvae smaller than about 6.5 mm SL, mechanical supports of fins and branchial arches were in a primordial stage of development. Supports and rays of the vertical fins and branchial arches rapidly developed from 6.5 mm SL, and all components appeared by about 10.5 mm SL. Thereafter body depth proportion changed and the supports and rays of the paired fins and gill-rakers developed. These developmental events were nearly or totally completed by about 17 mm SL, and we concluded that the larvae transformed to juveniles at this size. By this time, the mode of swimming of the fish shifted from undulating locomotion to caudal propulsion and that of feeding from swallowing paniculate food to filtering and concentrating substrate food matters using gill-rakers and the epibranchial organ. One of the most characteristic, and well-known, phenomena in the life history ofChanos chanos is the mass occurrence in the surf zone of postlarvae of a limited size range. In view of the scheme of the development of mechanical supports of the body and fins, they may acquire a swimming ability strong enough to move against the current only upon reaching about 10.5 mm SL, and if active shoreward migration of the larvae occurs, it is only during the late period of their journey from the spawning grounds to the shore. The sudden disappearance from the surf zone of larvae larger than 15–16 mm SL is obviously related to a change in food habit.     

Early life history of mebaru,sebastes inermis (scorpaenidae), in sendai bay, japan

The early life history of the viviparous scorpaenid,Sebastes inermis, in Sendai Bay, Japan, was studied and early development described. Newborn preflexion larvae ofS. inermis were about 5.2 mm BL. Notochord flexion occurred at 5.4–8.0 mm BL and transformation at 14–20 mm BL. Preflexion and flexion larvae ofS. inermis were distinguished from similar larvae by the pigmentation pattern along the dorsal and ventral midlines of the tail. Pigmentation inS. inermis was light throughout the larval and early juvenile periods. Planktonic larvae were particularly abundant in coastal waters of Sendai Bay but not offshore. Vertical and horizontal larval sampling indicated that early larvae occupied near surface waters and horizontal larval sampling indicated that early larvae shift to a benthic habitat occurred at about 12 mm BL, at the end of the postflexion larval period.Sebastes inermis do not have a distinct pelagic juvenile stage, unlike many North Pacific species ofSebastes.     

Luminous bacteria and light emitting fish: ultrastructure of the symbiosis

The luminescent fish Monocentris japonicus uses symbiotic luminous bacteria as a source of light. These bacteria live in light organs, complex tissue compartments, consisting of richly vascularized tubules or canals (in which the bacteria are cultured) lined with mitochondria-rich epithelial cells. The structure is consistent with a proposed model of symbiosis in which nutrients and oxygen are supplied by the vertebrate blood (vascular system). The nutrients, oxidized by the bacteria for growth and light production, are returned in part to the fish as pyruvate, which by reacting with mitochondrial oxygen regulates the light organ oxygen tensions. The luminous bacteria provide steady light that is modulated by passage through the melanocyte-containing dermis of the fish. Both the fish and the bacteria are highly adapted for their symbiotic coexistence.     

Growth and morphological development of laboratory-reared larval and juvenile Pangasianodon hypophthalmus

The morphological development, including the fins, body proportions and pigmentation, of laboratory-reared larval and juvenile Pangasianodon hypophthalmus was described and their behavioral features were observed under rearing conditions. Body lengths (BL) of larvae and juveniles were 3.0 ± 0.2 (mean ± SD) mm just after hatching, and 12.9 ± 1.1 mm on day 13, reaching 23.4 ± 1.8 mm on day 25 after hatching. Aggregate fin ray numbers (for caudal fin, principal soft ray number) attained their full complements in specimens larger than 12.8 mm BL. Notochord flexion began in yolksac larvae on day 0 (10.5 h after hatching), with teeth buds and barbels appearing with jaw formation in yolksac flexion larvae on day 1. Melanophores on the body increased with growth, with a broad vertical band forming on the lateral line and an oblique band extending from above the pectoral fin base towards the forepart of the anal fin during the postflexion larval and juvenile stages. Body proportions became relatively constant in juveniles, except for maxillary barbel length (MBL), which continued to decrease. Yolksac flexion larvae started feeding on day 2 with the onset of intense cannibalism. Yolks were completely absorbed by day 3, and cannibalism ended by day 6. Subsequently, fish displayed a schooling behavior with growth, preferring relatively dark areas during the juvenile stage.     

Early life history of kitsune-mebaru,sebastes vulpes (scorpaenidae), in the sea of japan

The larval and juvenile stages of kitsune-mebaru,Sebastes vulpes, based on 50 wild specimens collected in, the Sea of Japan, are described and illustrated, and some ecological aspects of the early life history (feeding, horizonal distribution and habitat shift) included. Preflexion larvae became extruded between 3.9–4.6 mm body length (BL) and notochord flexion occurred between 4.7–7.1 mm BL. Transformation from postflexion larvae to pelagic juventiles occurred between 13–17 mm BL. Compared with other rockfish species,S. vulpes is deep-bodied, throughout both larval and, juvenile stages. Larval and juvenileS. vulpes inhabit mainly coastal water surface layer (usually on the continental shelf), but do not occur offshore region (northwest of Oki Islands). Although someS. vulpes juveniles are associated with drifting seaweed, such clumps are not indispensable habitats for any stages. Surface-to-benthie migration of juveniles occurs at about 25 mm BL. Preflexion and flexion larvae feed mainly on copepod nauplii, and postflexion, transforming larvae and pelagic juveniles mainly on calanoid copepodites (Parracalanus parvus).     

Developmental morphology of the cyprinid fish Inlecypris auropurpureus

Embryonic, larval, and juvenile development of a Myanmarese cyprinid fish, Inlecypris auropurpureus, is described from laboratory-reared specimens. The eggs, measuring 0.9–1.0 mm in diameter, were demersal, almost spherical in shape, transparent and unpigmented, with a pale yellow yolk without oil globules. Hatching occurred 49–56 h after fertilization at 26.2°–27.3°C. The newly hatched larvae, measuring 2.9–3.1 mm in body length (BL) with 17 + 19–20 = 36–37 myomeres, had melanophores on the head and body. A cement organ on the forehead for adhering to objects during the yolk sac and early preflexion larval stages was distinctive. The yolk was completely absorbed at 3.6–4.0 mm BL. Notochord flexion was initiated at 5.1–5.6 mm BL and finished at 7.1 mm BL. Aggregate numbers of all fin rays were completed at 14 mm BL. Squamation was initiated midlaterally on the anterior trunk at 14 mm BL and completed at 27 mm BL. Although the eggs of I. auropurpureus resembled those of the closely related species Chela dadiburjori, Danio rerio, and Devario malabaricus, they differed from those of Danio rerio and Devario malabaricus in having a narrower perivitelline space. The larvae and juveniles of I. auropurpureus were also similar to those of C. dadiburjori, Danio rerio, and Devario malabaricus in general morphology, but they differed from the latter three species in having a series of dark blotches laterally on the body in the juvenile stage. Moreover, I. auropurpureus differed from C. dadiburjori in having more myomeres and a near-single row of melanophores on the body along the dorsal midline from the yolk-sac to early postflexion larval stages, from Danio rerio in having a cement organ on the forehead during the yolk-sac and early preflexion larvae, and a single melanophore on the lower eye margin in the early yolk-sac larvae, and from Devario malabaricus in having a single melanophore on the lower eye margin in the early yolk-sac larvae. The presence of a cement organ on the forehead indicates a close relationship among the genera Inlecypris, Chela, and Devario.