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.     

Seasonal profile of the diet of the dace Telestes karsticus Marčić & Mrakovčić, 2011 (Cyprinidae,Leuciscinae) endemic to Sušik Creek,Croatia

The first data on the feeding ecology of the endemic Telestes karsticus are presented for specimens sampled monthly (July 2007–July 2008) from the Su?ik Creek in the Lug Polje field, Croatia. Macrozoobenthos density was calculated and compared with the stomach contents of T. karsticus and the seasonal profile of the diet was examined. The ratio of gut length vs. standard length was 0.9 ± 0.12. Gut contents revealed that benthic invertebrates accounted for the majority of food consumed, although terrestrial arthropods, plant material, algae and fish were also found. Since this is the only fish species in the investigated creek, T. karsticus can be concluded to be cannibalistic. The proportion of empty guts found in all seasons was small, indicating year round feeding. The number and diversity of prey species was highest in spring and lowest in summer. Ivlev's electivity index was very high for Cladocera, Coleoptera, Trichoptera, Heteroptera and Hymenoptera in all seasons, suggesting positive selection for these prey groups, as opposed to Oligochaeta, Isopoda and Bivalva, which showed negative values in all seasons. For Actinopterygii, the Ivlev coefficient of selection was highly negative in spring, and highly positive in summer. Plant matter was detected in stomach contents in all seasons, with the highest share found in summer when other prey species were less available. The karstic dace is a generalised feeder, consuming the most available prey, and therefore can be classified as a euryphagous omnivore.     

Embryonic development of the threatened freshwater pipefish Microphis deocata (Hamilton, 1822)

The present study provides the first comprehensive embryonic development of the freshwater Syngnathid fish species, Microphis deocata (Hamilton), a Near Threatened pipefish endemic to the Brahmaputra River drainage in Northeast India and Bangladesh. Microphis deocata is a Gastrophori species as the males develop an abdominal brood pouch. Mature individuals were collected and maintained in well-aerated aquaria under controlled conditions to induce natural spawning. The number of eggs within the males' brood pouch ranged from 17 to 22 (for n = 10), measuring 0.7–1.0 mm in diameter. A total of 10 developmental stages could be recognized under four developmental periods namely, early embryogenesis, eye development, snout formation and juvenile. However, sensitivity, and therefore mortality, while handling of this species restricted the study from reporting the exact time intervals for stages following the blastodisc formation ~48 hr post fertilization. A newborn larvae measures ~14 mm and is free-swimming with distinct dorsal fin (with 31–32 rays) and a sector-shaped caudal fin (with 8–9 rays). The study aims to provide baseline information on the embryology of M. deocata in culture condition which will be helpful for future studies on conservation biology, population status and management of this species.     

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.     

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.     

Reproduction and early development of a freshwater pipefish <Emphasis Type="Italic">Microphis leiaspis</Emphasis> in Okinawa-jima Island,Japan

We investigated the size at maturation, breeding season, and morphological development of larvae and juveniles of a freshwater pipefish Microphis leiaspis, which belongs to Gastrophori, collected from three rivers on the northern part of Okinawa-jima Island, Japan. The minimum size of brooding males was 105–123 mm in standard length (SL). The smallest mature female was estimated to be ca. 130 mm SL from the analysis of gonadosomatic index (GSI) and histological observations of gonads. The breeding season was estimated to be from June to December according to monthly changes in female GSI, histological observations of gonads, and monthly changes in the occurrence of brooding males. The number of eggs in the male brood pouch ranged from 75 to 241 (mean ± SD: 152 ± 52, n = 22). The male releases newly hatched larvae in freshwater areas. After newborns grow in the sea, they return to freshwater areas of the rivers and attain maturity. Microphis leiaspis was conformed to have an amphidromous life history. Notochord length of the released larvae was 6.1 mm, with a well-developed finfold. Larvae attained 11.1 mm SL, formation of the caudal and dorsal fin rays was complete, and the caudal fin became lozenge shaped at 30 days after the release, and juveniles reached 36.0 mm SL at 63 days after release. In the period between 30 and 63 days after the release, formation of all fins except the pectoral fins was completed, and caudal fin rays were extended and sector shaped with deep slits between each fin ray. The morphology of the released larvae of M. leiaspis is similar to that of Gastrophori species, and the morphology of juveniles similar to other species of Microphis.     

Minute tubercles in bitterling larvae: Developmental dynamic structures to prevent premature ejection by host mussels

1. Bitterlings are small freshwater fish that use long ovipositors to lay eggs in host mussels, and they have morphological adaptations to increase larval survival. The most well‐known adaptation is the minute tubercles on the skin surface of larvae; they are developed in early‐stage larvae with weak swimming ability and disappear in free‐swimming larvae before they leave the host mussel.
2. In the present study, I comprehensively analyzed the developmental stages of Rhodeus pseudosericeus larvae, their morphological and physiological characteristics, their migration inside mussels, and the development of minute tubercle in order to elucidate the morphological function of the minute tubercles. These tubercles began to develop 1 day after hatching (formation stage), grew for 2–5 days (growth stage), reached the peak height after 6–7 days (peak stage), abruptly reduced in height after 8–10 days (abrupt reduction stage), and gradually reduced in height (reduction stage) until completely disappearing 27 days after hatching (disappearance stage).
3. The larvae remained in the mussels’ interlamellar space of the gill demibranchs until 10 days after hatching and began to migrate to the suprabranchial cavity 11 days after hatching. At this time, the larvae had a heart rate and the caudal fin began to develop. At 24 days after hatching, the minute tubercles had almost disappeared, and some individuals were observed swimming out of the mussels.
4. The results presented herein elucidate that the minute tubercles are the developmental dynamic structures that the bitterling larvae have morphologically adapted to prevent premature ejection from the mussel.

     

A new endemic cyprinid species from the Danube drainage

Populations of endemic Croatian dace were found to belong to two different species, one of which is first described in this study. Telestes karsticus sp. nov. differed morphologically from Telestes polylepis in the total count of lateral line scales, number of gill rakers and the shape of the posterior margin of the anal fin. Morphological differences were corroborated with mtDNA analyses (with p-distance between T. polylepis and T. karsticus sp. nov. ranging between 3·2 and 4·1%; and the number of substitutions between 37 and 47). The newly described species is geographically very localized. It has been recorded from only four localities around Velika Kapela and Mala Kapela mountains in Croatia.     

Adaptations to captive breeding of the longhorn beetle Morimus funereus (Coleoptera: Cerambycidae); application on amylase study

Captive breeding has been suggested as a method of conservation for many vertebrates, and is increasingly being proposed as a strategy for invertebrates. In this study, the growth, development and fertility of adults of the vulnerable cerambycid Morimus funereus reared in captivity are examined. Two oviposition cycles; from May to September and from January to March were studied and larvae from wild adults and from the progeny of captive adults (second generation larvae) were examined. Five to 12 instars were observed during larval development. Larval development was completed in 218 days (average) for the progeny of wild adults with an average mortality rate of 10.3% and in 226 days (average) for larvae from captive adults with mortality rate of 34.9%. First generation larval body weights were disparate during development, while second generation larvae had similar weights with no significant differences. In this study we have tested the potential of captive breaded M. funereus larvae as a model for investigation of digestive enzymes. Amylase from the midgut of larvae reared under laboratory conditions showed twofold higher specific activities with a decreased number of isoforms expressed, as compared to the enzyme from field-collected larvae. Captive breeding of M. funereus can be used in the future as a part of an effective conservation strategy for this rare insect species.     

Validation of daily increment formation in otoliths for Gymnocypris selincuoensis in the Tibetan Plateau,China

Daily increment validation in fish otolith is fundamental to studies on fish otolith microstructure, age determination and life history traits, and thus is critical for species conservation and fishery management. However, it has never been done for Schizothoracine fish, which is the dominant component of fish fauna in the Tibetan Plateau. This study validated the daily increment formation of Gymnocypris selincuoensis, as a representative of Schizothoracine fish, by monitoring the growth of hatchery‐reared larvae group and wild‐caught post‐yolk‐sac larvae group under controlled experiments. The results from monitoring the hatchery‐reared larvae group showed that sagittae and lapilli were found in yolk‐sac larvae, and formed 5–7 days before hatching, but asterisci were not found until 11 days post‐hatching. The first increment in sagittae and lapilli was formed in the first day after hatching. The daily periodicity of increment formation was examined and confirmed in sagittae and lapilli of both larvae groups. However, sagittae were better for age determination than lapilli for larvae at earlier days. For larval G. selincuoensis older than 50 days, lapilli were the only otolith pair suitable for larvae daily age determination. This study validated the daily increment formation in Schizothoracine fish for the first time has primary implications to other fishes from this subfamily.     

Evolution of slow and fast development in predatory ladybirds

The frequency distribution of the durations of development of 516 larvae of Adalia bipunctata is unimodal, and the fast‐ and slow‐developing larvae can be identified at the beginning of the fourth (=last) instar. To determine the advantages of fast and slow development, the survival, duration of development, growth and number of aphids consumed by fast‐ and slow‐developing fourth instar larvae fed different numbers aphids were recorded. The percentages of fast‐ and slow‐developing fourth instar larvae that survived when fed 0.5, 1 or an excess of aphids per day, surprisingly, did not differ. The slow‐developing larvae of both sexes took longer to complete their development than the fast‐developing larvae when fed 1 or an excess of aphids per day, and although the weights of the fast‐ and slow‐developing fourth instar larvae differed at the beginning of the instar, they did not differ at the end of this instar when fed 1 aphid per day. However, when reared on an excess of aphids per day, the adult weights of the fast‐developing individuals was greater than that of slow‐developing individuals. The average durations for which the larvae in the two groups survived when fed 0.5 aphids/day differed with the larvae of the fast‐developing individuals surviving for 9.8 ± 0.5 days and slow‐developing individuals 17 ± 1.3 days. Assuming that it is the rate of predator biomass increase, which is maximized by evolution, a model of the relationship between the rate of development/growth of a predator and that of its prey indicates that the optimum growth rate of a predator is positively associated with that of its prey. The evolutionary implications of these results are discussed.     

Embryonic development of the endangered Caspian brown trout,Salmo trutta caspius (Kessler, 1877)

The Caspian brown trout, Salmo trutta caspius is an endangered species of economic value and better knowledge of its biology will contribute to conservation measures. In the present study we characterized its early development to identify critical embryonic stages, by observing embryos and larvae from a broodstock maintained in captivity incubated at 9 ± 1°C. Morphological analysis of landmark stages of embryonic development revealed that first cleavage occurred at 189 degree‐h, a many‐celled blastoderm formed at 62 degree‐days (dd), a germ ring at 72 dd, the neural keel stage at 88 dd, the blastopore closed at 137 dd, the eye became pigmented at 188 dd and the larvae hatched at 373 dd. The detailed timing of these landmark features will help development of strategies aiming at increased aquaculture production of this species.     

Growth and morphological development of laboratory-reared larval and juvenile snakeskin gourami Trichogaster pectoralis

Morphological development, including that of fins, labyrinth organ, body proportions, and pigmentation, in laboratory-hatched larval and juvenile snakeskin gourami Trichogaster pectoralis is described. Body lengths (BL; mean ± SD) of larvae and juveniles were 2.3 ± 0.1 mm just after hatching (day 0) and 8.2 ± 0.6 mm on day 22, reaching 14.1 ± 2.3 mm on day 48. Aggregate fin ray numbers attained their full complements in juveniles >11.8 mm BL. Preflexion larvae started feeding on day 2 following upper and lower jaw formation, the yolk being completely absorbed by day 12. Subsequently, oblong conical teeth appeared in postflexion larvae >8.2 mm BL (day 16). Melanophores on the body increased with growth, with a large dark spot developing on the lateral midline at the caudal margin of the body in flexion larvae >6.1 mm BL. Subsequently, a broad vertical dark band from the eye to the caudal peduncle developed in postflexion larvae >8.9 mm BL. Proportions of head and pre-anal lengths became constant in postflexion larvae greater than ca. 9–10 mm BL, whereas those of maximum body depth, eye diameter, and snout length failed to stabilize in fish of the size examined in this study. First soft fin ray of the pelvic fin elongated, reaching over 40% BL. The labyrinth organ differentiated in postflexion larvae >7.4 mm BL (day 22). Comparisons of larval and juvenile morphology with another anabantoid species Anabas testudineus were also made, revealing several distinct differences, particularly in the numbers of myomeres and fin rays in the dorsal/anal fins, mouth location and body shape.     

Growth and morphological development of laboratory-reared larval and juvenile climbing perch <Emphasis Type="Italic">Anabas testudineus</Emphasis>

Morphological development, including fin and labyrinth organ, body proportions and pigmentation, in laboratory-reared larval and juvenile climbing perch Anabas testudineus was described and behavioral features under rearing condition were observed. Body lengths (BL) of larvae and juveniles were 1.9 ± 0.1 (mean ± SD) mm just after hatching (day-0), 8.7 ± 1.3 mm on day-19, reaching 18.4 ± 2.1 mm on day-35 after hatching. Aggregate fin ray numbers attained full complements in juveniles larger than 8.3 mm BL. Preflexion larvae started feeding on day-2 following formation of the upper and lower jaws, the yolk being completely absorbed by day-7 after hatching. Teeth appeared in flexion larvae larger than 5 mm BL on day-6, with cannibalism starting shortly after and continuing with further growth. Melanophores on the body increased with growth, a large dark spot developing on the lateral midline around caudal margin of the body in the postflexion and juvenile stages. The labyrinth organ differentiated in postflexion larvae larger than 7.2 mm BL on day-16, with air-breathing starting at the same time. Body proportions attained constant in postflexion larvae larger than 7.0 mm BL, and habitat of fish shifted from bottom to mid-layer. With the exception of fin ray numbers, the above morphological developments corresponded to behavioral shifts that occurred in the postflexion stage (ca. 7 mm BL), their subsequent continuity illustrating that the species possessed most juvenile-equivalent functions from ca. 7 mm BL.     

Improved genetic markers for monitoring recruitment dynamics in the endangered Mary River cod (Maccullochella mariensis)

The Mary River cod (Maccullochella mariensis) is a large predatory freshwater fish identified as a potential flagship for freshwater ecosystem conservation in Australia. The species is endemic to the Mary River catchment in eastern Australia, and is listed as Endangered. Previous conservation genetic assessment of Mary River cod was based on a small set of microsatellite loci developed for congeneric Maccullochella species. Here we develop a novel set of 15 microsatellite loci specific to M. mariensis, and demonstrate that these markers exhibit higher variability than those used previously. Mean number of alleles per locus was 4 and mean expected heterozygosity was 0.57. We genotyped 35 Mary River cod larvae belonging to a single cohort using the 15 novel loci and eight previously used loci, and found 10 full‐sib family groups along with clear genetic differentiation between individuals collected from the two sub‐catchments – Tinana Creek and Mary River. Microsatellites presented here will be useful for cost‐effective monitoring of genetic diversity and recruitment dynamics in this endangered fish species.     

Morpho‐histological and ultra architectural changes during early development of endangered golden mahseer Tor putitora

Ultrastructural and histological changes in the embryonic and larval surface during ontogenesis of the endangered golden mahseer Tor putitora is studied here for the first time. Embryonic development was completed 91–92 h after fertilization at an ambient temperature of 23° ± 1° C (mean ± s.d. ). The gastrula stage was characterized by presence of the Kupffer's vesicle, notochord, ectoderm and endoderm cells. Primordial germ cells were clearly identifiable from c. 55 h post‐fertilization at the organogenesis stage. Mean total length of newly hatched larvae was 7·0 ± 0·5 mm. Scanning electron microscopy of newly hatched larvae demonstrated vitelline arteries, microridged epithelial cells and mucous gland openings over much of the body surface. Eye, oral cavity, pharyngeal arches, heart, intestinal loop, prosencephalon, cephalic vesicle and nasal epithelium were clearly distinguished in 3 day old hatched individuals. In 6 day old individuals, caudal‐fin rays and internal organs were evident. The dorsal fin became prominent at this stage and larvae began swimming at the surface. The reserved yolk material was totally absorbed 8–11 days after hatching and larvae began feeding exogenously. Tor putitora exhibited a longer early developmental period than other cyprinids reared at similar temperatures.     

An update on the status of native and exotic freshwater fishes of Italy

An updated, corrected checklist of both native and established alien freshwater fish species in Italy is given based on molecular, morphological and biogeographical data. Some 12 native species, reported as conspecific with transalpine species in official Italian ichthylogical literature, are in fact endemics. Previous taxonomic confusion has resulted in the introduction of several alien species, either with official stockings or mixed in as impurities. Rehabilitated species include the cyprinids Scardinius hesperidicus, Scardinius scardafa and Telestes savigny from northern Italy, as well as Squalius ruffoi and the Telestes comes from southern Italy. Squalius albus is a junior synonym of S. squalus. The endemic gudgeon, previously assigned to the genus Romanogobio, is returned to the genus Gobio (G. benacensis). Phoxinus lumaireul is a junior synonym of P. phoxinus. Among the Salmonidae, Salmo cenerinus is a junior synonym of S. marmoratus, while Salmo farioides represents the trout species of the Adriatic lineage for which a neotype is designated. Thymallus aeliani represents the endemic lineage of grayling of the Adriatic populations. The esocid Esox cisalpinus is an endemic pike species and Esox flaviae is a junior synonym; the extensive exportation as well as the presence of this species throughout Europe is possibly due to humans. Among sculpins, Cottus scaturigo and C. ferrugineus are junior synonyms of C. gobio. The International Union for Conservation of Nature (IUCN) categories for native species of Italy are updated. At present, 52 native freshwater fish species are listed: 2 are extinct (Acipenser sturio and Huso huso), 12 are critically endangered, 7 endangered, 10 vulnerable, 3 near‐threatened, 15 low concern and 3 data‐deficient; 35 species are the result of human transfers. Among the 51 introduced species, 6 are recently established (Leuciscus leuciscus, Oreochromis niloticus, Poecilia reticulata, Xiphophorus helleri, Amatitlania nigrofasciatus, Hemichromis sp.), 37 are already established, 5 are probably established and 3 are non‐established Chinese carp, maintained in the wild by intensive stockings. The family most involved is the Cyprinidae, with 22 alien and 20 native species.     

Release of planula larvae, settlement and development of Siderastrea stellata Verrill, 1868 (Anthozoa, Scleractinia)

We herein provide the first observations on planulation, larval development, and metamorphosis of Siderastrea stellata, an endemic reef-building species that occurs along the northeastern and southeastern coasts of Brazil. The release and settlement of larvae were observed in two distinct periods. The first started 3 days after collection on January 28 during the last quarter, whereas the second started 2 months later, on April 10 during the change from new moon to the first quarter. Planulation continued throughout approximately 48 h. Brooded larvae released from the mouth contained zooxanthellae, and underwent settlement after 48 h. Newly extruded larvae stayed in close contact with parental polyps. Fusion was observed among larvae from the same colony. The first septal cycle was formed by day 2–3, while the third cycle of exosepta became evident 15 days after protosepta development. The development of the corallite of primary polyp was slow, and after 9 months of analyses no evidence of budding was obtained, this indicating that colonial development is likely to be a late event in the S. stellata life history.     

Erythritol ingestion impairs adult reproduction and causes larval mortality in Drosophila melanogaster fruit flies (Diptera: Drosophilidae)

Previous feeding studies showed the polyalcohol erythritol was toxic when ingested by adult laboratory fruit flies (Drosophila melanogaster). We asked whether erythritol could additionally affect fly population growth either through larval toxicity or through effects on adult reproduction. Females did not avoid laying on food substrates with 1M erythritol; laying rate on 1M erythritol food was similar to control food when females were given free‐choice access. Eggs laid or placed on 0.5 M to 2.5 M erythritol foods hatched at normal rates, suggesting erythritol was not toxic to eggs upon contact. Drosophila melanogaster larvae readily consumed food containing 1 M erythritol, but none of these larvae reached pupation. Longevity of larvae feeding on in 1 M erythritol food was significantly reduced relative to controls, and mean ± SE larval lifespan on erythritol was 1.54 ± 0.10 days (max. = 3 days). Exposing cohorts of second‐instar larvae to food with varying concentrations of erythritol showed the LD50 (at 24 hr) concentration was approximately 0.6 M. Taken together, these results suggest erythritol could be employed in effective larval‐sink baits. Adults flies fed with erythritol produced significantly fewer eggs on days when they fed on 1 M erythritol, and egg production was significantly reduced for one additional day after the adults were moved to control food. These findings suggest erythritol is rapid and effective at temporarily suppressing D. melanogaster reproduction, increasing its potential for use in effective insect population control.