Early Postlarval Stages Research Articles

Ontogenetical development of branchial chambers of Litopenaeus vannamei (Boone, 1931) and theirinvolvement in osmoregulation: ionocytes and Na+/K+-ATPase.

Branchial chambers constitute the main osmoregulatory site in almost all decapod crustaceans. However, few studies have been devoted to elucidate the cellular function of specific cells in every osmoregulatory structure of the branchial chambers. In decapod crustaceans, it is well-known that the osmoregulatory function is localized in specific structures that progressively specialize from early developmental stages while specific molecular mechanisms occur. In this study, we found that although the structures developed progressively during the larval and postlarval stages, before reaching juvenile or adult morphology, the osmoregulatory capabilities of Litopenaeus vannamei were gradually established only during the development of branchiostegites and epipodites, but not gills. The cellular structures of the branchial chambers observed during the larval phase do not present the typical ultrastructure of ionocytes, neither Na+/K+-ATPase expression, likely indicating that pleura, branchiostegites, or bud gills do not participate in osmoregulation. During early postlarval stages, the lack of Na+/K+-ATPase immunoreactivity of the ionocytes from the branchiostegites and epipodites suggests that they are immature ionocytes (ionocytes type I). It could be inferred from IIF and TEM results that epipodites and branchiostegites are involved in iono-osmoregulation from PL15, while gills and pleura do not participate in this function.

Microbial community composition associated with early developmental stages of the Indian white shrimp, Penaeus indicus.

Gut microbiota is known to influence the physiology, health, nutrient absorption, reproduction, and other metabolic activities of aquatic organisms. Microbial composition can influence intestinal immunity and are considered as health indicators. Information on gut microbial composition provides potential application possibilities to improve shrimp health and production. In the absence of such information for Penaeus indicus, the present study reports the microbial community structure associated with its early developmental stages. Bacterial community associated with the early developmental stages (egg, nauplii, zoea, mysis, PL1, PL6 and PL12) from two hatchery cycles were analysed employing 16S rRNA high throughput sequencing. Proteobacteria and Bacteroidetes, were the two dominant phyla in P. indicus development stages. Sequential sampling revealed the constant change in the bacterial composition at genus level. Alteromonas was dominant in egg and nauplii stage, whilst Ascidiaceihabitans (formerly Roseobacter) was the dominant genera in both PL6 and PL12. The bacterial composition was highly dynamic in early stages and our study suggests that the mysis stage is the critical phase in transforming the microbial composition and it gets stabilised by early post larval stages. This is the first report on the composition of microbiota in early developmental stages of P. indicus. Based on these results the formation of microbial composition seems to be influenced by feeding at early stages. The study provides valuable information to device intervention strategies for healthy seed production.

A case study on the distribution of the environmental resistome in Korean shrimp farms

Hundreds of tons of antibiotics are widely used in aquaculture to prevent microbial infections and promote fish growth. However, the overuse of antibiotics and chemical products can lead to the selection and spreading of antibiotic-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs), which are of great concern considering the threat to public health worldwide. Here, in-depth metagenome sequencing was performed to explore the environmental resistome and ARB distribution across farming stages in shrimp farms and examine anthropogenic effects in nearby coastal waters. A genome-centric analysis using a metagenome binning approach allowed us to accurately investigate the distribution of pathogens and ARG hosts in shrimp farms. The diversity of resistomes was higher in shrimp farms than in coastal waters, and the distribution of resistomes was dependent on the farming stage. In particular, the tetracycline resistance gene was found mainly at the early post-larval stage regardless of the farm. The metagenome-assembled genomes of Vibrio spp. were dominant at this stage and harbored tet34, which is known to confer resistance to oxytetracycline. In addition, opportunistic pathogens such as Francisella, Mycoplasma, Photobacterium, and Vibrio were found in abundance in shrimp farms, which had multiple virulence factors. This study highlights the increased resistance diversity and environmental selection of pathogens in shrimp farms. The use of environmental pollutants on farms may cause an increase in resistome diversity/abundance and the transmission of pathogens to the surrounding environment, which may pose future risks to public health and aquatic organisms.

Probiotics, lactic acid bacteria and bacilli: interesting supplementation for aquaculture

Probiotics administration in aquafeed is known to increase feed consumption and absorption due to their capacity to release a wide range of digestive enzymes and nutrients which can participate in digestion process and feed utilization, along with the absorption of diet components led to an increase in host's health and well-being. Furthermore, probiotics improve gut maturation, prevention of intestinal disorders, predigestion of antinutrient factors found in the feed ingredients, gut microbiota, disease resistance against pathogens and metabolism. The beneficial immune effects of probiotics are well established in finfish. However, in comparison, similar studies are less abundant in the shellfish. In this review, the discussions will mainly focus on studies reported the last 2years. In recent studies, native probiotic bacteria were isolated and fed back to their hosts. Although beneficial effects were demonstrated, some studies showed adverse effects when treated with a high concentration. This adverse effect may be due to the imbalance of the gut microbiota caused by the replenished commensal probiotics. Probiotics revealed greatest effect on the shrimp digestive system particularly in the larval and early post-larval stages, and stimulate the production of endogenous enzymes in shrimp and contribute with improved the enzyme activities in the gut, as well as disease resistance.

Skeletal muscle protease activities in the early growth and development of wild Atlantic salmon (Salmo salar L.).

Growth-related dynamics of intracellular protease activities in four year classes of the Atlantic salmon (Salmo salar L. 1758) parr and smolts inhabiting salmon rivers of northwestern Russia (the White Sea basin) were studied. Cathepsin B, cathepsin D, proteasome, and calpain activities in the skeletal muscles of salmon were assessed to investigate their relative contribution to the total protein degradation as well as to young fish growth process. It was confirmed that calpain activity dominates in salmon muscles while proteasome plays a minor role, in contrast to terrestrial vertebrates. Calpain and proteasome activities were maximal at the early post-larval stage (in parrs 0+) and declined with age (parrs 1+ through 2+) dropping to the lowest level in salmon smolts. Annual growth increments and proteolytic activities of calpains and proteasome in the muscles of salmon juveniles changed with age in an orchestrated manner, while lysosomal cathepsin activities increased with age. Comparing protease activities and growth increments in salmon parr and smolts we suggested that the partial suppression of the protein degradation could be a mechanism stimulating efficient growth in smoltifying salmon. Growth and smoltification-related dynamics of protease activities was quite similar in salmon populations from studied spawning rivers, such as Varzuga and Indera; however, some habitat-related differences were observed. Growth increments and protease activities varied in salmon parr 0+ (but not on later ages) inhabiting either main rivers or small tributaries apparently due to habitat difference on the resources for fish growth.

Remarkable vertical shift in residence depth links pelagic larval and demersal adult jellynose fish

Deep-sea fish show diverse migratory behaviors across depths at different life stages. The historical residence depths of jellynose fish Ateleopus japonicus and Ijimaia dofleini (Ateleopodidae) were reconstructed from otolith microstructures and isotopic compositions. δ18O values in the otolith core areas ranged from −0.5 to −1.3‰ among individuals, suggesting that larval and post-larval stages lived in the mixed layer (50–200m). Otolith growth increment widths surged for 10–30 rings around 300–600µm from the core, indicating a fast-growth phase during the early post-larval stage. Fish then migrated downward to 350–800m depth at about 2 months of age, possibly during the post-larval metamorphosis to the juvenile. Otolith growth increments became narrower and otolith δ13C values increased from −5 to −1‰, suggesting a lower growth and metabolic rate when the fish experienced colder water during the downward migration. After arrival at the deepest waters, the fish then migrated upward to the continental margin or upper slopes where the adults persistently resided. A translucent otolith zone was formed after the residence depth shift from the deepest waters to shallower depths, indicating a transition from pelagic to bathydemersal life on the continental shelf or break. The down-and-up shift in residence depth of jellynose fish represents an indirect settlement process to the adult residence depth, which might be associated with a unique post-larval stage moving offshore before the downward migration. The results filled the gap of vertical distributions of jellynose fish from pelagic larvae near the sea surface to the bathydemersal adult dwelling on the continental shelf break.

Effect of the early temperature on the growth of larvae and postlarvae turbot, Scophthalmus maximus L.: muscle structural and ultrastructural study.

Turbot specimens were kept at three temperatures (T s ): warm (W) (21-22°C), ambient (A) (17-18°C) and cold (C) (13-14°C) during the larval and early postlarval stages. At 90days posthatching (dph), all of them were transferred to ambient T until 190dph. At 2-3dph, the specimens showed a monolayer of red muscle and immature white fibres; external or dermomyotome cells (presumptive myogenic cells) were observed on the surface of the red muscle. In the following stages, many myogenic cells and presumptive myogenic precursors were observed within the myotome, presumably derived of the dermomyotome. When comparing the growth at the same age (2, 10, 25, 37dph), the body length and the muscle growth were positively influenced by the warm T, being the hyperplasia the muscle parameter more significantly influenced. The development rate was also positively correlated with the high T: the beginning of the metamorphosis took place at 15, 23 and 25dph at W, A and C temperatures, respectively, with the highest body length values at ambient temperature. The metamorphosis finished at 25, 30 and 37dph at W, A and C temperatures, respectively, with the highest body length values at warm temperature. However, the muscle cellularity was similar in all the groups at the end of the metamorphosis. At 90 and 190dph, the largest body length was observed at W temperature. However, the muscle cellularity was similar between A and W; the number of fibres was similar in all the groups at 190dph, which shows the beginning of a compensatory muscle growth in A and C, mainly in A.

Presumed postlarval pentacrinoids from the Lower Devonian Hunsrück Slate, Germany

Several tiny crinoids with crowns as small as 1 mm, or less, in width are newly recognized from the Hunsruck Slate of southwestern Germany. The presence of erect arms above an amorphous calyx in some specimens can be inferred. Based on comparison with the size and gross morphology of developmental stages in living crinoids, these tiny Hunsruck crinoids are judged to be at an early postlarval stage that is analogous to the pentacrinoid stage just after development from the stalked, but armless, smaller cystidean larval stage found in both living comatulids and isocrinids. Some of these tiny crinoids have a stalk up to 4 mm long attached to a now pyritized former substrate. Their clustered occurrence suggests gregarious settlement of larvae. Taxonomic identification of these presumed pentacrinoids is not possible, even to the sub-class level, although they are preserved with larger juveniles of the cladids Propoteriocrinus and Lasiocrinus. These larger juveniles exhibit 3-D pyritized calcite plates, whereas the probable pentacrinoids appear to be preserved as flattened, micro-crystalline pyritized dermal tissues that enclosed lightly calcified, porous ossicles. The pentacrinoids were likely buried within weeks or months of hatching, based on developmental stages in similar-sized living crinoids. These tiny crinoids, presumably pentacrinoids, are a further example of the extraordinarily detailed preservation of delicate tissues in pyrite from the Hunsruck Slate. They are most likely the pentacrinoid stage from one or more of the crinoid taxa (30 genera) present in the Hunsruck Slate. Assuming these are not microcrinoids, they are the first report of pentacrinoids from the fossil record and document that a Palaeozoic sister group to modern crinoids had similar developmental stages.

The genome and occlusion bodies of marine Penaeus monodon nudivirus (PmNV, also known as MBV and PemoNPV) suggest that it should be assigned to a new nudivirus genus that is distinct from the terrestrial nudiviruses.

BackgroundPenaeus monodon nudivirus (PmNV) is the causative agent of spherical baculovirosis in shrimp (Penaeus monodon). This disease causes significant mortalities at the larval stage and early postlarval (PL) stage and may suppress growth and reduce survival and production in aquaculture. The nomenclature and classification status of PmNV has been changed several times due to morphological observation and phylogenetic analysis of its partial genome sequence. In this study, we therefore completed the genome sequence and constructed phylogenetic trees to clarify PmNV’s taxonomic position. To better understand the characteristics of the occlusion bodies formed by this marine occluded virus, we also compared the chemical properties of the polyhedrin produced by PmNV and the baculovirus AcMNPV (Autographa californica nucleopolyhedrovirus).ResultsWe used next generation sequencing and traditional PCR methods to obtain the complete PmNV genome sequence of 119,638 bp encoding 115 putative ORFs. Phylogenetic tree analysis showed that several PmNV genes and sequences clustered with the non-occluded nudiviruses and not with the baculoviruses. We also investigated the characteristics of PmNV polyhedrin, which is a functionally important protein and the major component of the viral OBs (occlusion bodies). We found that both recombinant PmNV polyhedrin and wild-type PmNV OBs were sensitive to acid conditions, but unlike the baculoviral OBs, they were not susceptible to alkali treatment.ConclusionsFrom the viral genome features and phylogenetic analysis we conclude that PmNV is not a baculovirus, and that it should be assigned to the proposed Nudiviridae family with the other nudiviruses, but into a distinct new genus (Gammanudivirus).Electronic supplementary materialThe online version of this article (doi:10.1186/1471-2164-15-628) contains supplementary material, which is available to authorized users.

Colonization patterns of decapods into artificial collectors: seasonality between habitat patches

Understanding colonization of new habitats by recruits is essential to comprehend local abundance patterns of adult populations. Colonization of benthic habitats by new colonizers is achieved through the influx of larvae from the water column and movements from adjacent habitats. Colonization, i.e., the composition and abundance of new colonizers, into a focal habitat may be affected by surrounding habitats. In this study, we investigated seasonal variation through an annual cycle in postlarval settlement patterns of decapod crustaceans colonizing artificial collectors on different habitats, in particular: Cymodocea nodosa seagrass meadows, sandy patches, ‘urchin-grazed’ barrens and macroalgal-dominated beds, which are arranged in patches within seascapes in the eastern Atlantic. Our results showed that: 1) decapods, at least some species, settle preferentially on artificial collectors deployed on vegetated substrates when compared to those deployed on unvegetated substrates; 2) despite most decapods have a planktonic larval phase with wide oceanic dispersal, the presence of crustaceans in particular habitats suggests a great specificity at their early post-larval stages; and 3) although subtropical waters of Gran Canaria have a weak seasonal variation, early postlarval stages of decapod crustaceans showed two distinct settlement periods, i.e., a ‘cold’ and a ‘warm’ season.

Direct evidence for maternal inheritance of bacterial symbionts in small deep-sea clams (Bivalvia: Vesicomyidae)

Bacterial symbiont transmission is a key step in the renewal of the symbiotic interaction at each host generation, and different modes of transmission can be distinguished. Vesicomyidae are chemosynthetic bivalves from reducing habitats that rely on symbiosis with sulfur-oxidizing bacteria, in which two studies suggesting vertical transmission of symbionts have been published, both limited by the imaging techniques used. Using fluorescence in situ hybridization and transmission electron microscopy, we demonstrate that bacterial symbionts of Isorropodon bigoti, a gonochoristic Vesicomyidae from the Guiness cold seep site, occur intracellularly within female gametes at all stages of gametogenesis from germ cells to mature oocytes and in early postlarval stage. Symbionts are completely absent from the male gonad and gametes. This study confirms the transovarial transmission of symbionts in Vesicomyidae and extends it to the smaller species for which no data were previously available.

Uneven abundance of the invasive sun coral over habitat patches of different orientation: An outcome of larval or later benthic processes?

Larval behavior in the water column and preference among natural benthic habitats are known to determine initial spatial distribution patterns in several sessile marine invertebrates. Such larval attributes can be adaptive, promoting adult benthic distributions which maximize their fitness. Further benthic processes may, however, substantially change initial distribution of settlers. In this study, we first characterized spatial distributions of adult colonies and single-polyp recruits of the invasive azooxanthellate coral Tubastraea coccinea over substrates of different orientation, and evaluated their consistency at both small (several tens of meters) and intermediate (a few km) spatial scales. We then assessed, through field and laboratory experiments, larval preferences and relative settlement and recruitment rates on surfaces with different orientations to determine whether processes taking place during the larval and early post-larval stages could help explain the distribution patterns of recruits and adult colonies. Results suggest that larval passive buoyancy and active larval behavior, unrelated to light conditions, determine a clear settlement distribution pattern, in which the density of settlers is highest at undersurfaces and almost nil at upward facing horizontal substrates. Except for an almost absence of settlers, recruits and adult individuals on upward facing horizontal habitat, there is substantial mismatch between the distribution of settlers and that of recruits and adult colonies. The latter were also common in vertical substrate in the field. We speculate that coastal runoff at the study area and subsequent sedimentation may inhibit coral development on flat upward facing habitat, and that competitive interference and pre-emptive interactions with other azooxanthellate corals could constrain abundance of T. coccinea in underface horizontal habitat.

Post-Embryonic Transcriptomes of the Prawn Macrobrachium rosenbergii: Multigenic Succession through Metamorphosis

Like many metazoans, the freshwater prawn Macrobrachium rosenbergii begins its post-embryonic life with a set of morphologically distinct planktonic larval stages, followed by a benthic post-larval stage during which the maturing organism differs from the larvae both ecologically and physiologically. Understanding of the molecular basis underlying morphogenesis in crustaceans is limited to the observation that methyl farnesoate, the non-epoxidated form of the insect juvenile hormone, acts as the active crustacean juvenoid. Molt steroids were also linked to morphogenesis and several other molecular pathways, such as Hedgehog and Wnt, are known to underlie morphogenesis in all metazoans examined and, as such, are thought to do the same in crustaceans. Using next generation sequencing, we deep-sequenced the transcriptomes of several larval and post-larval stages. De novo assembly, followed by bioinformatics analysis, revealed that many novel transcripts are over-expressed in either larvae- or post-larvae-stage prawn, shedding light on the molecular basis underlying M. rosenbergii metamorphosis. Fast larval molting rates and periodic morphological changes were reflected in over-expression of transcripts annotated to the cell cycle, DNA replication and morphogenic pathways (i.e., Hedgehog and Wnt). Further characterization of transcripts assigned to morphogenic pathways by real-time RT-PCR reconfirmed their over-expression in larvae, albeit with a more complex expression pattern when examined in the individual developmental stages. The expression level of an orthologue of cytochrome P450, 15A1, known to epoxidize methyl farnesoate in insects, was increased in the late larval and early post-larval stages, in accordance with the role of methyl farnesoate in crustacean metamorphosis. This study exemplifies the applicability of a high-throughput sequencing approach for studying complex traits, including metamorphosis, providing new insight into this unexplored area of crustacean research.

Pyrosequencing of Haliotis diversicolor Transcriptomes: Insights into Early Developmental Molluscan Gene Expression

BackgroundThe abalone Haliotis diversicolor is a good model for study of the settlement and metamorphosis, which are widespread marine ecological phenomena. However, information on the global gene backgrounds and gene expression profiles for the early development of abalones is lacking.Methodology/Principal FindingsIn this study, eight non-normalized and multiplex barcode-labeled transcriptomes were sequenced using a 454 GS system to cover the early developmental stages of the abalone H. diversicolor. The assembly generated 35,415 unigenes, of which 7,566 were assigned GO terms. A global gene expression profile containing 636 scaffolds/contigs was constructed and was proven reliable using qPCR evaluation. It indicated that there may be existing dramatic phase transitions. Bioprocesses were proposed, including the ‘lock system’ in mature eggs, the collagen shells of the trochophore larvae and the development of chambered extracellular matrix (ECM) structures within the earliest postlarvae.ConclusionThis study globally details the first 454 sequencing data for larval stages of H. diversicolor. A basic analysis of the larval transcriptomes and cluster of the gene expression profile indicates that each stage possesses a batch of specific genes that are indispensable during embryonic development, especially during the two-cell, trochophore and early postlarval stages. These data will provide a fundamental resource for future physiological works on abalones, revealing the mechanisms of settlement and metamorphosis at the molecular level.

Ontogenetic changes of innate immune parameters from eggs to early postlarvae of white shrimp Litopenaeus vannamei (Crustacea:Decapoda)

To date, little is known about the immune system during early ontogeny for crustaceans with high commercial value, such as white shrimp (Litopenaeus vannamei). The aim of this study was to characterize several immunological parameters during ontogeny (from eggs to early postlarvae) of L. vannamei under farming conditions. Phenoloxidase activity showed the highest peak 3.6±0.4×10−2U μg−1 at the first nauplius stage, followed by a continuous decrease until dropping to 14% of the peak value in early postlarval (PL) stages. Superoxide dismutase activity reached the highest peak 9.9±0.3×10−3Uμg−1 after metamorphosis to the protozoea stage, then to half that value after metamorphosis to mysis stage and kept dropping until reaching a minimum of 0.5±0.4×10−3Uμg−1 in PL stages. In larval stages peroxidase activity showed a maximum of 2.5±0.4×10−6Uμg−1 after metamorphosis to mysis, and a gradual increase was observed in PL stages until attaining the highest value 4.6±0.8×10−6Uμg−1. Generation of superoxide anion showed a progressive increase after spawning in an evident relationship with growth, reaching the highest value of 7.0±0.4×10−6Uμg−1 in PL-7. To our knowledge, this is the first study that characterizes immunological parameters through the first stages of ontogeny of L. vannamei under farming conditions.

Toward a sustainable production of genetically improved all-male prawn (Macrobrachium rosenbergii): Evaluation of production traits and obtaining neo-females in three Indian strains

Sex reversal technology, realized through androgenic gland (AG) manipulation, was recently introduced as a process for production of all-male producing broodstock. This technology exploits however, a relatively small number of sex-reversed broodstock. Thus, both genetic improvement via a breeding program and prevention of inbreeding are needed to ensure the sustainability of such technology. Three wild strains of prawns originating from geographically (though not necessarily genetically) isolated locations in India [Gujarat (G), Kerala (K) and West Bengal (WB)] were assessed for their suitability as breeders for all-male production. In addition, their potential for a selective breeding program was evaluated. A comparative evaluation of early sex segregation, sex reversal, growth performance, and population structure in the three selected strains was performed. Among the purebred strains, after eight months of grow out in earthen ponds, growth performance of the WB strain was the best (59.39±1.08g), while that of G was the poorest (26.50±0.94g). Strain-additive genetic effects for body weight at harvest were highest for the WB strain (+45.9%) and lowest for the G strain (−28.3%). Body masses of WB×K and WB×G crosses were 14.2% and 8.8% above the mean mass of the purebred strains, respectively, while that of the K×G cross was 23% below this value. In most crosses, males reached heavier mean body weights than did females with higher frequencies of the large male morphotypes being seen in the WB purebred strain and its respective crosses. Reciprocal effects for body mass ranged from 4% to 14.9% below the mean of the purebred strains. These negative signs mean that in the two crosses involving the WB strain, growth performance is higher when this stain was used as the sire strain. Similarly, the growth performance of the K×G cross was higher when the former was used as the sire strain. Average heterosis effect for body weight was minor (−0.51±0.73) and did not differ significantly from zero. The high correlation between strain additive effects (the major source of variation in growth) and total performance for body weight (r=0.927) indicate the existence of valuable genetic variation that could be exploited in a selective breeding program. For all-male production, males from the three strains were segregated at early post-larval stages and microsurgical AG removal was performed. In all the strains, similar low levels of complete sex reversal into functional neo-females (genetic males) were realized (0.17% −0.34%). These produced relatively small numbers of neo-female to be crossed with normal males to produce the desired all-male population, but raise the possibility that such a process could result in a genetic bottleneck. Thus, a genetic improvement scheme for each strain integrated with periodical crosses of the resulting neo-females from one strain with males from another strain is suggested to avoid inbreeding.

Triploid Black Tiger shrimp ( Penaeus monodon) performance from egg to harvest age

In this study we induced triploids in a single family of Black Tiger shrimp ( Penaeus monodon) using chemical shock induction to stop the second meiotic division. The family was reared from egg to harvest age (post-larval stage 184) in a combination of commercial hatchery and experimental controlled environment tank rearing systems. During the rearing phase ploidy, survival, weight, sex ratio and gonad morphology of sibling diploids and induced triploids were periodically assessed. This is the first published report of the culture performance of triploid P. monodon. The induction rate of triploids at the nauplii stage was 83.4 ± 0.04% but this proportion had dropped to 62.45% by the time shrimp metamorphosed to the protozeal stage. Proportions of triploids remained fairly constant throughout the remaining larval and early post-larval stages (ranging from 60.47% to 72.38%), but dropped again to 56.32% by the time shrimp reached harvest age. The reduction in triploid rate from egg to harvest age is thought to be due to a combination of factors including heightened sensitivity of triploids to continual handling and general lower survival of triploids compared to diploids. Once shrimp reached harvest age diploid females were significantly heavier (42.95 ± 2.43 g, P < 0.05) than all other categories of shrimp. There was no significant difference in weight ( P > 0.05) between triploid females, diploid males and triploid males (35.14 ± 2.26 g, 34.02 ± 0.89 g and 31.79 ± 1.56 g respectively). The final sex ratio of harvest age triploid P. monodon was 1 female:1.625 males. Histological examination of harvest age female and male gonads revealed a drastic reduction in the development of both ova and sperm in triploid shrimp compared to diploids. The limited numbers of oocytes and sperm produced in triploids failed to fully mature by harvest age. No sperm were observed within spermatophores of triploids at harvest age when a gross morphological examination was performed and no sperm bundles were seen in the vas deferens. Using the same examination technique, diploid harvest age males had sperm within the spermatophores and vas deferens, and sperm bundles. Results indicate that reproductive sterility is likely for both female and male triploid P. monodon.

Effect of biofloc technology (BFT) on the early postlarval stage of pink shrimp Farfantepenaeus paulensis: growth performance, floc composition and salinity stress tolerance

Biofloc rearing media provides a potential food source for shrimp reared in limited or zero water exchange systems. This culture system is environmentally friendly as it is based on limited water use and minimal effluent is released into the surrounding environment. In this study, we evaluated the survival, growth performance and salinity stress tolerance of pink shrimp Farfantepenaeus paulensis postlarvae reared from PL10 to PL25 in a biofloc technology limited water exchange system. PL (mean ± SD weight and length of 14 ± 10 mg and 8.10 ± 0.7 mm, respectively) were reared in nine 40-L plastic tanks with a stocking density of 10PL/L. Three culture treatments were applied (1) culture in the presence of bioflocs and commercial feed supply (FLOC + CF); (2) culture in the presence of biofloc without feed supply (FLOC) and (3) culture in clear water with feed supply (control). Final biomass and survival were significantly higher in FLOC + CF treatment than the control (P 0.05) between treatments was found for salinity tolerance over 24 and 48 h durations. The proximate analysis of floc shown high levels of crude protein (30.4%), but low levels of crude lipids (0.5%). The continuous availability of bioflocs had a significant effect on growth and survival of F. paulensis postlarvae cultured in BFT nursery systems.

Infection of adult migratory river shrimps, Macrobrachium ohione, by the branchial bopyrid isopod Probopyrus pandalicola

Abstract. Macrobrachium ohione is a migratory (amphidromous) river shrimp (Decapoda, Caridea) that may be parasitized by the branchial parasite Probopyrus pandalicola (Isopoda, Bopyridae). The parasite disrupts gonadal maturation and spawning in female shrimps, resulting in the total loss of reproduction. Shrimps are usually infected by bopyrid parasites during the late zoeal or early postlarval stages; in this study, we investigated the apparent parasite infection of adult shrimps. We analyzed the relationships between parasite body size (total length) and host shrimp body size (carapace length) to test the hypothesis that parasite infection of adult shrimps occurs during the shrimps' reproductive migrations. The results presented here indicate that infection of adult shrimps is common in M. ohione in the Atchafalaya and Mississippi Rivers, Louisiana, USA. In the two upriver sites sampled, Butte La Rose (BLR) and River Bend (RB), parasite size was not associated with host body size. In these locations, many parasitized adult M. ohione were infected with immature P. pandalicola (40.3% in BLR and 51.2% in RB), indicating that the shrimps were adults at the time of infection. A possible explanation is that when female shrimps enter the estuary to hatch larvae, they molt and spawn another brood. The smaller male shrimps that accompany the females downstream are also assumed to molt and continue growth. The intermediate host of the parasite is an estuarine copepod, and thus the parasite cryptoniscus larva that infects the host shrimp is primarily estuarine as well. Newly molted shrimps have soft cuticles, which may facilitate their infection by parasite cryptonisci. Our conclusion is that most infections of adult shrimps occur during their migration into estuarine waters, the primary habitat of infective parasite larvae, and that host vulnerability is probably increased following host ecdysis.

Artemia replacement in co-feeding regimes for mysis and postlarval stages of Litopenaeus vannamei: Nutritional contribution of inert diets to tissue growth as indicated by natural carbon stable isotopes

The nutritional contribution from co-fed Artemia nauplii and inert diets to growth in mysis and early postlarval shrimp Litopenaeus vannamei was assessed by analyzing the carbon stable isotope ratios ( δ 13C) in diets and shrimp tissue. Artemia nauplii and inert diets showed significantly different δ 13C values but similar carbon contents and were supplied as single diets and also in three co-feeding regimes in which 25, 50 and 75% of the Artemia was replaced by inert diet on a dry weight basis, so that all feeding regimes provided similar amounts of dietary carbon. Shrimp δ 13C values were significantly influenced by the different feeding regimes and reached isotopic equilibrium with their respective diets as soon as 5 days. Survival was significantly higher in co-fed animals than in those fed either Artemia or inert diet alone. There was no significant difference in growth between shrimp fed on Artemia only and those co-fed Artemia with inert diet, although the variability was high. Growth and survival were very low in shrimp fed only inert diet. Results from an isotope mixing model suggest that observed nutrient contributions from Artemia nauplii were significantly higher than expected contributions indicated by proportions established in the co-feeding regimes. From mysis 3 to postlarvae 5, nutrient contributions to growth in the dietary regime providing equal carbon amounts of each diet type ranged from 73 to 87% for Artemia and from 13 to 27% for inert diet. Shrimp fed the 25% Artemia replacement regime exhibited a significantly higher retention of dietary carbon from the inert diet than those fed inert diet alone. This may have resulted from greater ingestion and/or assimilation of the inert diet in the presence of Artemia, combined with the higher growth rate supported by co-feeding. The results demonstrate the effectiveness of up to 50% replacement of Artemia with inert diet for L. vannamei mysis and early postlarval stages, indicating also that the inert diet may provide specific nutrients that promote higher survival, while digestibility may limit its contribution to tissue growth. The isotopic influence of maternal nutrients and dietary sources on earlier larval stages was also described.