Extracellular vesicles (EVs) released by cells play a role in intercellular communication. Reporter and targeting proteins can be modified and exposed on the surface of EVs to investigate their half-life and biodistribution. A characterization of membrane-bound Gaussia luciferase (mbGluc) revealed that its signal was detected also in a form smaller than common EVs (<70 nm). We demonstrated that mbGluc initially exposed on the surface of EVs, likely undergoes proteolytic cleavage and processed fragments of the protein are released into the extracellular space in active form. Based on this observation, we developed a new assay to quantitatively track shedding of membrane proteins from the surface of EVs. We used this assay to show that ectodomain shedding in EVs is continuous and is mediated by specific proteases, e.g. metalloproteinases. Here, we present a novel tool to study membrane protein cleavage and release using both in vitro and in vivo models.
The complete life cycle of a pennellid copepod Peniculus minuticaudae Shiino, 1956 is proposed based on the discovery of all post-embryonic stages together with the post-metamorphic adult females infecting the fins of threadsail filefish Stephanolepis cirrhifer (Monacanthidae) cultured in a fish farm at Ehime Prefecture, Japan. The hatching stage was the infective copepodid. The life cycle of P. minuticaudae consists of six stages separated by moults: the copepodid, four chalimi and adult. In this study, the adult males were observed frequently in precopulatory amplexus with various stages of females however, copulation occurs only between adults. Fertilized pre-metamorphic adult females carrying spermatophores may detach from the host and settle again before undergoing massive differential growth into the post-metamorphic adult female. Comparison of the life cycle of P. minuticaudae has been made with three known pennellids: Lernaeocera branchialis (Linnaeus, 1767), Cardiodectes medusaeus (Wilson, 1908) and Lernaeenicus sprattae (Sowerby, 1806). Among the compared species, P. minuticaudae is the first ectoparasitic pennellid to be discovered to complete its life cycle on a single host without any change in infection site preferences between infective copepodid and fertilized pre-metamorphic female.
The presently reported study provides a detailed morphological description of the female and the male of a new species of the genus Parabrachiella-Parabrachiella jarai sp. nov. The parasites were sampled from marine fish, silver sillago, Sillago sihama (Perciformes: Sillaginidae), captured in Malaysia in 1994 and Hong Kong in 1995. The new species bears some resemblance to Parabrachiella lata (Song et Chen, 1976) but differs from it in details of second antenna, mandible, and maxilliped. The genus Parabrachiella currently covers 67 species including those recently transferred from Neobrachiella Kabata, 1979. An amended generic diagnosis is proposed for Parabrachiella and Thysanote. Some members of Parabrachiella are herewith transferred to Thysanote and some Thysanote are now placed in Parabrachiella.
The effects of salinity, temperature, and light conditions on the reproduction and development of harpacticoid copepod, Nitocra affinis f. californica under controlled laboratory conditions were determined. Seven different salinity levels (5, 10, 15, 20, 25, 30, 35 ppt), four temperatures (20, 25, 30, 35 degrees C), three different light intensities (25, 56, 130 micromol m(-2) s(-1)) and photoperiods (24 h:0 h, 1 h:23 h, 12 h:12 h LD cycle) were employed in this study. The highest (p < 0.05) overall reproduction and fastest development time were achieved by copepods reared under 30-35 ppt salinity. The optimum temperature required for the maximum reproduction was 30 degrees C while under 30 degrees C and 35 degrees C the copepod development time was shortest (p < 0.05) compared to other temperature levels. The overall reproduction was highest (p < 0.05) and development rate of N. affinis was shortest (p < 0.05) under lowest light intensity (25 micromol m(-2) s(-1)). Continuous light (24 h:0 h LD) inhibited the egg production while, continuous darkness (1 h:23 h LD) and 12 h:12 h LD significantly favoured the overall reproductive activity of the female. Photoperiods 1 h:23 h and 12 h:12 h LD yielded highest total (p < 0.05) offspring female(-1) coupled with highest (p < 0.05) survival percentage. This study illustrated that although N. affinis can tolerate wide range of environmental conditions, prolonged exposure to subnormal environments affect its reproduction and development. This study showed that this species can be mass cultured for commercial purposes and has a potential to be used for toxicity studies due to its high reproductive performance fast development and a wide range of tolerance to environmental conditions.
The Caligidae is a family of parasitic copepods containing over 30 recognised genera. They are commercially important parasites as they cause disease in numerous finfish aquaculture facilities globally. Morphological features are used to distinguish between the genera and Pseudocaligus has traditionally been differentiated from Caligus solely by the presence of a much reduced form of the fourth thoracic leg. Currently there are numerous DNA sequences available for Caligus spp. but only the type species, Pseudocaligus brevipedis, has molecular data available, so systematic studies using molecular phylogenetic analyses have been limited.
A microsporidian hyperparasite, Desmozoon lepeophtherii, of the parasitic copepod Lepeophtheirus salmonis (salmon louse), infecting farmed Atlantic salmon (Salmo salar), was first discovered in the west of Scotland in 2000. Heavily infected salmon lice are easily recognised as they have large opaque inclusions distributed throughout the body. The prevalence of salmon lice with visible signs of microsporidiosis can be up to 10% of the population from certain farm sites. The microsporidian was also isolated from the host Atlantic salmon suggesting it may have a two host life cycle. The authors believe that the infection in immunocompetent salmon may be latent, becoming acute during periods of infection with another pathogen or during sexual maturation. Since its first discovery in Scotland, Desmozoon lepeophtherii has been subsequently reported from Norway, and more recently from the Pacific coast of North America.
Three strains of the chain-forming diatom Skeletonema marinoi, differing in their production of polyunsaturated aldehydes (PUA) and nutritional food components, were used in experiments on feeding, egg production, hatching success, pellet production, and behavior of three common planktonic copepods: Acartia tonsa, Pseudocalanus elongatus, and Temora longicornis. The three different diatom strains (9B, 1G, and 7J) induced widely different effects on Acartia tonsa physiology, and the 9B strain induced different effects for the three copepods. In contrast, different strains induced no or small alterations in the distribution, swimming behavior, and turning frequency of the copepods. 22:6(n-3) fatty acid (DHA) and sterol content of the diet typically showed a positive effect on either egg production (A. tonsa) or hatching success (P. elongatus), while other measured compounds (PUA, other long-chain polyunsaturated fatty acids) of the algae had no obvious effects. Our results demonstrate that differences between strains of a given diatom species can generate effects on copepod physiology, which are as large as those induced by different algae species or groups. This emphasizes the need to identify the specific characteristics of local diatoms together with the interacting effects of different mineral, biochemical, and toxic compounds and their potential implications on different copepod species.
This study was aimed at determining the optimum temperature for culturing the copepod, Euterpina acutifrons. The trial was conducted for 10 days in chambers at temperatures of 25⁰C, 27⁰C, 29⁰C and 31⁰C. Ten adult individuals of the copepod were randomly collected and placed into three replicate experimental flasks for each treatment. Throughout the trial, the salinity, light intensity, and photoperiod were maintained at 30 ±2psu, 100molm-2s-1 and 12:12 light-dark cycle, respectively. The copepods were fed with 80,000cell/ml Isochrysis sp. daily. At the end of the trial, the total numbers of E. acutifrons nauplii, copepodites and adults were determined and counted using Sedgwick-Rafter. The highest population was found at 27⁰C with mean total population of 800±100 individuals from an initial of 10 individuals. This was followed by those reared at 25⁰C and 29⁰C where the population counts were 700±100 individuals and 367±115 individuals, respectively. At the 31⁰C, all the copepod specimens were found dead on day 5th. Statistical analysis showed that the temperature had a significant effect (P
This study focuses on the parasites in Dascyllus trimaculatus fish in Bidong Island. Though D. trimaculatus is a common reef fish, there is lack on parasites studies of this fish in Malaysia. The objectives of this study are to identify metazoan parasites and analyse the prevalence and mean intensity the parasites in D. trimaculatus. Sampling was randomly conducted on a coral colony of 100 meters within a 15m depth at Pantai Pasir Cina,BidongIsland.ThisresearchresultedintheidentificationofacopepodLernaeocera branchialis and a nematode from the family Camallanidae. The copepods collected in this study infected 20 out of 42 D. trimaculatus specimens, whereas the nematode-like parasites collected infected 18 of out of 42 D. trimaculatus. All copepods found in the gills of D. trimaculatus had a prevalence of 4.95 and mean intensity of 47.62%, while the nematode-like parasite, which were found in the brain, had 2.72 prevalence and 42.86% mean intensity. This elucidates that both parasites are categorised as having light levelbut common frequency of infection, and the current IUCN status of D. trimaculatus has not been reported to be harmedby parasites. Though the mean intensity of both parasites isnot harmful, the prevalence are concerning and might increase in the future, with further incorporation and monitoring of climate change factors that may affect the damselfish.
Cyclopoid copepod has a potential as live feed that can provide ornamental fish larvae with energy and essential nutrients, and promote their growth and survival, as well as affect their colouration. However, the nutrition, growth and nutritional requirements of this species are poorly understood. This research focuses on comparing the efficacy of enrichment types on copepods towards the growth rate, survival rate and colouration of Pterophyllum scalare larvae. The enrichment of copepods consists of four enrichment procedures (rice bran, palm kernel cake (PKC),Chlorella and yeast). This study was carried out over 50 days and the growth rate, survival rate and colouration (growth rate, survival rate, SGR and colouration) were used to evaluate the responses of P. scalare larvae towards different enrichment methods. Specific growth rates (8.0161 ± 1.4928 %; P = 0.775) and survival rates (66.667 ± 5.337%; P = 0.815) of fish larvae were found to not be significantly different (P = 0.775, P = 0.815; P >0.05). However, the colouration of the fish larvae was affected by the types of enrichment used (P
Pseudo-nitzschia nanaoensis sp. nov. is described from waters around Nan'ao Island (South China Sea), using morphological data and molecular evidence. This species is morphologically most similar to P. brasiliana, but differs by a denser arrangement of fibulae, interstriae, and poroids, as well as by the structure of the valvocopula and the narrow second band. Pseudo-nitzschia nanaoensis constitutes a monophyletic lineage and is well differentiated from other species on the LSU and ITS2 sequence-structure trees. Pseudo-nitzschia nanaoensis makes up the basal node on the LSU tree, and forms a sister clade with a group of P. pungens and P. multiseries on the ITS2 tree. The ability of cultured strains to produce domoic acid was assessed, including its possible induction by the presence of a copepod and brine shrimp, by liquid chromatography-tandem mass spectrometry. However, no strains showed detectable domoic acid.
Copepoda is one of the most ecologically important animal groups on Earth, yet very few genetic resources are available for this Subclass. Here, we present the first whole genome sequence (WGS, acc. UYDY01) and the first mRNA transcriptome assembly (TSA, Acc. GHAJ01) for the tropical cyclopoid copepod species Apocyclops royi Until now, only the 18S small subunit of ribosomal RNA gene and the COI gene has been available from A. royi, and WGS resources was only available from one other cyclopoid copepod species. Overall, the provided resources are the 8th copepod species to have WGS resources available and the 19th copepod species with TSA information available. We analyze the length and GC content of the provided WGS scaffolds as well as the coverage and gene content of both the WGS and the TSA assembly. Finally, we place the resources within the copepod order Cyclopoida as a member of the Apocyclops genus. We estimate the total genome size of A. royi to 450 Mb, with 181 Mb assembled nonrepetitive sequence, 76 Mb assembled repeats and 193 Mb unassembled sequence. The TSA assembly consists of 29,737 genes and an additional 45,756 isoforms. In the WGS and TSA assemblies, >80% and >95% of core genes can be found, though many in fragmented versions. The provided resources will allow researchers to conduct physiological experiments on A. royi, and also increase the possibilities for copepod gene set analysis, as it adds substantially to the copepod datasets available.
Snapper had been cultured in Malaysia since 1980 due to the fry availability and the high demand. However, details on the caligids infestation were not properly documented. This study was carried out to determine the prevalence, mean intensity and site preference of Caligus rotundigenitalis (Caligidae, Siphonostomatoida) a parasitic copepod on cage cultured crimson snapper, Lutjanus erythropterus from Bukit Tambun, Penang, Malaysia. A total of 70 specimens of cultured snapper were examined based on different infestation sites such as head, body as well as operculum. The specimens were separated into three groups according to the size of the fish. C. rotundigenitalis was found to be the only species infesting L. erythropterus with the prevalence and the mean intensity of 81.4% and 5.6±4.4, respectively. There was a significant difference between the prevalence of site infestation of the body and inner operculum sites. The prevalence of C. rotundigenitalis was highest on inner operculum of the fish followed by the body and head. However, there was no significant difference in the distribution of C. rotundigenitalis over the different infestation sites derived from the three groups. The information obtained from this study can be used for more effective control measures of ectoparasitic copepod infestation in floating cages.
The Asian seabass is euryhaline, therefore it is interesting to describe the infestation and survival of caligids at varying salinity on the host. In this study, two different brackish water culture systems with monoculture and polyculture practices were investigated for the occurrence of Caligus spp. on Lates calcarifer. Polyculture practices mainly consisted of snapper (Lutjanus spp.), grouper (Epinephelus spp.) and seabass (L. calcarifer), while the monoculture was stocked with only seabass. A total of 777 Caligus spp. specimens were isolated from the sampling in 2009, consisting of three species; Caligus chiastos, Caligus epidemicus and Caligus rotundigenitalis. In 2011, the total specimen was increased to 3110 and two additional species were found; Caligus punctatus and one unknown species (Caligus sp.). A 98.6% of the total examination was represented by C. epidemicus. Constant presence of C. epidemicus was observed throughout the study, regardless the differences in between culturing practices and systems. This species was able to survive within wide salinity range, from 5 to 28 ppt. The other isolated species (C. chiastos, C. punctatus, C. rotundigenitalis and Caligus sp.) were only found infesting in polyculture cages with the salinity ranging from 25 to 28 ppt. Despite accounts for less than 2% of the total specimens, these species may able to produce a challenge for L. calcarifer polyculture farming activity due to their capability for host switching. The present study revealed the potential risk for cross-species transmission in polyculture practices.
Heat shock response (HSR), in terms of transcription regulation of two heat shock proteins genes hsp70 and hsp90), was analysed in a widespread tropical copepod Pseudodiaptomus annandalei. The mRNA transcripts of both genes were quantified after copepods at a salinity of 20 underwent an acclimation process involving an initial acclimation temperature of 29 °C, followed by gradual thermal ramping to the target exposure temperature range of 24-36 °C. The respective cellular HSR and organismal metabolism, measured by respiratory activity at exposure temperatures, were compared. The fold change in mRNA expression for both hsp70 and hsp90 (8-9 fold) peaks at 32 °C, which is very close to 32.4 °C, the upper thermal optimum for respiration in the species. Unexpectedly, the modelled HSR curves peak at only 3 °C (hsp90) and 3.5 °C (hsp70) above the mean water temperature (29.32 °C) of the copepod in the field. We propose that copepods in tropical waters adopt a preparative HSR strategy, early at the upper limit of its thermal optimum, due to the narrow thermal range of its habitat thus precluding substantial energy demand at higher temperatures. However, the model suggests that the species could survive to at least 36 °C with short acclimation time. Nevertheless, the significant overlap between its thermal range of hsp synthesis and the narrow temperature range of its habitat also suggests that any unprecedented rise in sea temperature would have a detrimental effect on the species.
Background. Climate change concurrent with anthropogenic disturbances can initiate serial changes that reverberate up the food chain with repercussions for fisheries. To date, there is no information available concerning the combined effects of global warming and human impacts on tropical marine food webs. While temperate copepods respond differently to warming and environmental stressors, the extent to which tropical copepods can adapt to rising temperature of already warm waters remains unknown. We hypothesize that sea warming and other anthropogenic disturbances over the long term will have the greatest impact on the copepod community in nearshore waters where their effects are accentuated, and therefore vulnerable and resilient species could be identified. Methods. Zooplankton samples were collected during two time periods (1985-86 and 2014-15) interposed by marked anthropogenic disturbances, and at the same five stations located progressively from inshore to offshore in Klang Strait, Malaysia, following the asymmetrical before-after-control-impact (BACI) design. Copepods were identified to species, and results were interpreted by univariate (ANOVA) and multivariate (PERMANOVA, PCO) analyses of the computed species abundance and diversity measures. Results. Copepod total abundance was not significantly different among stations but higher after disturbance than before disturbance. However, changes in the abundance of particular species and the community structure between time periods were dramatic. Coastal large-bodied calanoid species (e.g., Acartia spinicauda, Calanopia thompsoni, Pseudodiaptomus bowmani and Tortanus forcipatus) were the most vulnerable group to disturbance. This however favored the opportunistic species (e.g., Oithona simplex, O. attenuata, Hemicyclops sp., Pseudomacrochiron sp. and Microsetella norvegica). Small-bodied copepods (e.g., Paracalanus sp., Parvocalanus crassirostris and Euterpina acutifrons) were unaffected. Centropages tenuiremis was likely an introduced species. There was no significant loss in species richness of copepods despite the dramatic changes in community structure. Discussion. Sea warming and other human-induced effects such as eutrophication, acidification and coastal habitat degradation are likely the main factors that have altered copepod community structure. The large-bodied estuarine and coastal calanoid copepods are surmised to be vulnerable to eutrophication and hypoxia, while both resilient and opportunistic species are largely unaffected by, or adaptable to, degraded coastal environments and observed sea surface temperature (SST) rise. It is forecasted that SST rise with unmitigated anthropogenic impacts will further reduce large-bodied copepod species the favoured food for fish larvae with dire consequences for coastal fish production.
The possible links between river flow, zooplankton abundance and the responses of zooplanktivorous fishes to physico-chemical and food resource changes are assessed. To this end, the seasonal abundance, distribution and diet of the estuarine round-herring Gilchristella aestuaria and Cape silverside Atherina breviceps were studied in the Kariega Estuary. Spatio-temporal differences were determined for selected physico-chemical variables, zooplankton abundance and zooplanktivorous fish abundance and distribution. Results indicated that, following a river flood event in winter (>30 m3 s-1 ), altered physico-chemical conditions occurred throughout the estuary and depressed zooplankton stocks. Abundance of G. aestuaria was highest in spring, with this species dominant in the upper and middle zones of the estuary, while A. breviceps was dominant in summer and preferred the middle and lower zones. The catch per unit of effort of both zooplanktivores also declined significantly following the flooding, thus suggesting that these fishes are reliant on zooplankton as a primary food source for healthy populations. Copepods dominated the stomach contents of both fish species, indicating a potential for strong interspecific competition for food, particularly in the middle reaches. Temporal differences were evident in dietary overlap between the two zooplanktivorous fish species and were correlated with river flow, zooplankton availability and fish distribution. The findings of this study emphasize the close trophic linkages between zooplankton and zooplanktivorous fishes under changing estuarine environmental conditions, particularly river flow and provide important baseline information for similar studies elsewhere in South Africa and the rest of the world.
At the present time, no artificial larval diet is capable of entirely fulfilling the dietary requirements of several larval fish and crustacean species. Zooplankton live food is the basic foundation of fish larviculture, and successful rearing of fish larvae still heavily depends on an adequate supply of nutritious live food. Despite being important, the production protocols of copepods and cladocerans (Moina) are still underdeveloped in hatcheries. Rotifers and Artemia are the most commonly used live foods. However, these live foods are evidently lacking in crucial nutrient constituents. Hence, through nutrient enrichment, live food with the nutritional profile that meets the requirements of fish larvae can be produced. With the aim to maximize the effectiveness of production to optimize profitability, it is important to evaluate and improve culture techniques for the delivery of micro- and macro-nutrients as feed supplements to larvae in aquaculture systems. Bioencapsulation and enrichment are the evolving techniques in aquaculture that are commonly employed to enhance the nutritional quality of live food by integrating nutrients into them, which subsequently improves the growth, survival, and disease resistance of the consuming hosts. This review aims to highlight some of the approaches and methods used to improve the nutritional quality of live food by modifying their nutrient composition, which could have immense promise in the enhancement of aquatic animal health.
The lagoon of Setiu Wetlands has high biodiversity of bivalve’s species. The majority of villagers are fisherman and bivalves are one of their income-generating activities. Studies on parasites of bivalves are important to our knowledge for maintenance of natural resources. This study investigated the parasite prevalence among four commercially exploited shellfish species from Setiu Wetland, Malaysia. A total of 120 samples were collected during the low tide time for four shellfish species which were Polymesoda expansa, Meretrix meretrix, Anadara sp. and Crassostrea iredalei. The bivalve specimens were measured on its length with and without shell, width, height, weight and microscopically examined on the presence of parasites focused at gills, muscle and digestive tract. The discovered macroparasite were fixed into 70% ethanol solution for preservation. In this study, Anadara sp. was highest infestation of parasites, while Polymesoda expansa has the least infestation of parasites. There were occurrence of copepod and Nematopsis sp. in Anadara sp., M. meretrix and C. iredalei. Apart from that, there were other parasites observed in four bivalve species such as metacercaria, cestode larvae, Panopeus sp., Pinnotheres sp., Balanus sp. and unidentified ciliates. Ecological factors, feeding activity, season and abundance of definitive host were known to be elements that altered the prevalence of parasites in host. Upon the observation, a high prevalence of Nematopsis sp. were observed in three species of bivalves except P. expansa. Whereas for other parasites were occurred in low prevalence and intensity, without causing significant damage towards host.
The development of novel mosquito control tools is a key prerequisite to build effective and reliable Integrated Vector Management strategies. Here, we proposed a novel method using cigarette butts for the synthesis of Ag nanostructures toxic to young instars of the malaria vector Anopheles stephensi, chloroquine (CQ)-resistant malaria parasites Plasmodium falciparum and microbial pathogens. The non-target impact of these nanomaterials in the aquatic environment was evaluated testing them at sub-lethal doses on the predatory copepod Mesocyclops aspericornis. Cigarette butt-synthesized Ag nanostructures were characterized by UV-vis and FTIR spectroscopy, as well as by EDX, SEM and XRD analyses. Low doses of cigarette butt extracts (with and without tobacco) showed larvicidal and pupicidal toxicity on An. stephensi. The LC50 of cigarette butt-synthesized Ag nanostructures ranged from 4.505 ppm (I instar larvae) to 8.070 ppm (pupae) using smoked cigarette butts with tobacco, and from 3.571 (I instar larvae) to 6.143 ppm (pupae) using unsmoked cigarette butts without tobacco. Smoke toxicity experiments conducted against adults showed that unsmoked cigarette butts-based coils led to mortality comparable to permethrin-based positive control (84.2 and 91.2%, respectively). A single treatment with cigarette butts extracts and Ag nanostructures significantly reduced egg hatchability of An. stephensi. Furthermore, the antiplasmodial activity of cigarette butt extracts (with and without tobacco) and synthesized Ag nanostructures was evaluated against CQ-resistant (CQ-r) and CQ-sensitive (CQ-s) strains of P. falciparum. The lowest IC50 values were achieved by cigarette butt extracts without tobacco, they were 54.63 μg/ml (CQ-s) and 63.26 μg/ml (CQ-r); while Ag nanostructure IC50 values were 72.13 μg/ml (CQ-s) and 77.33 μg/ml (CQ-r). In MIC assays, low doses of the Ag nanostructures inhibited the growth of Bacillus subtilis, Klebsiella pneumoniae and Salmonella typhi. Finally, the predation efficiency of copepod M. aspericornis towards larvae of An. stephensi did not decrease in a nanoparticle-contaminated environment, if compared to control predation assays. Overall, the present research would suggest that an abundant hazardous waste, such as cigarette butts, can be turned to an important resource for nanosynthesis of highly effective antiplasmodials and insecticides.