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Concept: Cyprinidae

165

BACKGROUND: The risks of fish-borne zoonotic trematodes (FZT) to human health constitute an important problem in Vietnam. The infection of humans with these trematodes, such as small liver trematodes (Clonorchis sinensis and Opisthorchis viverrini), intestinal trematodes (Heterophyidae) and others is often thought to be linked to fish culture in areas where the habit of eating raw fish is common. Juvenile fish produced in nurseries are often heavily infected with FZT and since fishes are sold to aquaculture facilities for growth, control of FZT in these fishes should be given priority. Controlling the first intermediate host (i.e., freshwater gastropods), would be an attractive approach, if feasible. The black carp, Mylopharyngodon piceus, is a well-known predator of freshwater snails and is already used successfully for biological control of snails in various parts of the world including Vietnam. Here we report the first trials using it for biological control of intermediate host snails in nursery ponds stocked with 1-week old fry (10–12 mm in length) of Indian carp, Labeo rohita. METHODS: Semi-field and field experiments were set up to test the effect of black carp on snail populations. In the semi-field experiment a known quantity of snails was initially introduced into a pond which was subsequently stocked with black carp. In the field trial in nursery ponds, density of snails was estimated prior to a nursing cycle and at the end of the cycle (after 9 weeks). RESULTS: The results showed that black carp affect the density of snail populations in both semi-field and field conditions. The standing crop of snails in nursery ponds, however, was too high for 2 specimens to greatly reduce snail density within the relatively short nursing cycle. CONCLUSIONS: We conclude that the black carp can be used in nursery ponds in Northern Vietnam for snail control. Juvenile black carp weighing 100 - 200g should be used because this size primarily prey on intermediate hosts of FZT and other studies have shown that it does not prey on fish fry of other species. It may be necessary to use a high stocking density of black carp or to reduce snail density in the nursery ponds using other measures (e.g. mud removal) prior to stocking fry in order for the black carp to keep the density of intermediate host snails at a very low level.

Concepts: Digenea, Carp, Cyprinidae, Aquaculture, Snail, Gastropoda, Red River Delta, Black carp

100

Both endotherms and ectotherms (e.g., fish) increase their body temperature to limit pathogen infection. Ectotherms do so by moving to warmer places, hence the term “behavioral fever.” We studied the manifestation of behavioral fever in the common carp infected by cyprinid herpesvirus 3, a native carp pathogen. Carp maintained at 24°C died from the infection, whereas those housed in multi-chamber tanks encompassing a 24°C-32°C gradient migrated transiently to the warmest compartment and survived as a consequence. Behavioral fever manifested only at advanced stages of infection. Consistent with this, expression of CyHV-3 ORF12, encoding a soluble decoy receptor for TNF-α, delayed the manifestation of behavioral fever and promoted CyHV-3 replication in the context of a temperature gradient. Injection of anti-TNF-α neutralizing antibodies suppressed behavioral fever, and decreased fish survival in response to infection. This study provides a unique example of how viruses have evolved to alter host behavior to increase fitness.

Concepts: Immune system, Gene expression, Bacteria, Infection, Thermoregulation, Fever, Cyprinidae, Goldfish

29

Jatropha curcas seeds are rich in oil and protein. The oil is used for biodiesel production. Jatropha seed cake (JSC) obtained after oil extraction is rich in protein; however, it is toxic (phorbol esters content 1.3 mg/g) and consists of 50-60% shells, which are indigestible. The principle of isoelectric precipitation was used to obtain Jatropha protein isolate (JPI) from JSC and it was detoxified (DJPI). Carp (n = 45, 20.3 ± 0.13 g) were randomly distributed into five groups with three replicates and for 12-week fed iso-nitrogenous diets (crude protein 38%): Control [fishmeal (FM)-based protein]; J(50) and J(75) (50% and 75% of FM protein replaced by DJPI); S(50) and S(75) (50% and 75% of FM protein replaced by soy protein isolate). Growth performance and nutrient utilisation parameters were highest in S(75) group and not significantly different to those in J(50) and S(50) groups but were significantly higher than those for all other groups. Similar trend was observed for protein and energy digestibilities of experimental diets, whereas opposite trend was observed for the feed to gain ratio. Activities of intestinal digestive enzymes did not different significantly between the five groups. In conclusion, DJPI is a good quality protein source for carp.

Concepts: Soybean, Koi, Common carp, Carp, Cyprinidae, Cyprinus, Jatropha, Biodiesel

28

Objective  To evaluate the physiological effect and response to noxious stimulation at five concentrations of MS-222 in koi (Cyprinus carpio). Study design  Prospective experimental study. Animals  Twenty-one healthy adult unknown sex koi fish weighing mean 450 ± SD 120 g. Methods  Each fish was exposed to five different concentrations of MS-222 (50, 70, 110, 150 and 190 mg L(-1) ) in a random sequence during the same anaesthetic event. For each concentration of MS-222, vital functions such as heart rate (HR) (via Doppler) and opercular rate (OpR) were recorded after a standardized induction period. Response to two noxious stimuli in the form of haemostat clamp pressure applied on the tail and the lip was evaluated, and blood was drawn to measure biochemical and blood gas values. Results  Decrease in response to noxious stimulation with an increase of MS-222 concentration both for the lip (p = 0.0027) and the tail (p < 0.0001) stimulus was observed. Biochemical values were unaffected by the concentration of MS-222 with the exception of lactate concentration which was weakly correlated with the duration of anaesthesia (r = 0.31, p < 0.001) and the number of times the fish was clamped or bled prior to sampling (r = 0.23, p < 0.001). Opercular rate decreased with the increase in anaesthetic concentration, and HR was not affected. Conclusions and clinical relevance  Our results indicated a decrease in response to stimulus and a decrease in OpR that were associated with increased concentrations of MS-222. This may assist in establishing anaesthetic protocols using MS-222 in fish and supports the use of supramaximal pressure stimuli to teleost fish under variable MS-222 concentrations as a model for future studies.

Concepts: Measurement, Koi, Common carp, Carp, Cyprinidae, Cyprinus, Goldfish, Cypriniformes

28

Silver carp, Hypopthalmichthys molitrix is one of the most economically valuable fish species in Bangladesh. However, its production is often hindered by parasite-induced mortality. The present study reports the intensity of parasitic infestation in 216 specimens of H. molitrix collected from different fish markets in Rajshahi City, Bangladesh. Nine different parasite species (Trichodina pediculatus, Dactylogyrus vastator, Ichthyophthirius multifilis, Gyrodactylus elegans, Lernaea sp., Apiosoma sp., Myxobolus rohitae, Camallanus ophiocephali, and Pallisentis ophiocephali) were recovered from the gill, skin, stomach, and intestine of host fish. The highest level of infection was observed for host skin, while lower levels were observed for host gill, stomach, and intestine. The results also revealed that the intensity of parasite infection in different organs of H. molitrix varied with the season. In particular, the highest levels of infection were recorded during the winter period (November-February), when fish are most susceptible to parasites. The findings of the study will help in the management and conservation of H. molitrix.

Concepts: Immune system, Bacteria, Parasitism, Carp, Cyprinidae, Organs, Hypophthalmichthys, Silver carp

28

Abstract Spring viremia of carp virus (SVCV) causes a highly contagious and serious disease of freshwater cyprinid fishes, generating significant economic and ecological impacts throughout the world. The SVCV is therefore listed as a notifiable pathogen by the International Organization for Animal Health. In June 2011, a significant mortality event of wild common carp Cyprinus carpio occurred in Minnehaha Creek near its confluence with Mississippi River Pool 2 in Minneapolis, Minnesota. Clinical signs of moribund fish included hemorrhagic lesions in the skin, eyes, and internal adipose tissue. The SVCV was isolated from pooled kidney and spleen of the fish. Rhabdovirus particles were seen upon examination of infected cell culture fluid by electron microscopy. The virus was confirmed to be SVCV subtype Ia by reverse transcription PCR and sequencing. This is the first report of SVCV within the state of Minnesota and the ninth documented case in North America. Received March 27, 2012; accepted July 2, 2012.

Concepts: Koi, Common carp, Carp, Cyprinidae, Goldfish, Mississippi River, Minneapolis, Mississippi National River and Recreation Area

24

Members of Candidia, Nipponcrysis, Parazacco, Opsariichthys, and Zacco are a group of East Asian freshwater fishes, and these five genera form a monophyletic group within the subfamily Oxygastrinae (Cypriniformes: Cyprinidae). In this study, we first determined complete mitogenomes of Candidia pingtungensis, Opsariichthys acutipinnis, O. chengtui, and Zacco acanthogenys. The four mitogenomes with the length of 16 611-16 615 bp displayed the same patterns in gene arrangements and the use of start and stop codons for protein-coding genes. Our phylogeny divided Candidia, Nipponcrysis, Parazacco, Opsariichthys, and Zacco into two major groups that the former three genera consisted of a group and the latter two genera formed another group. The phylogeny also revealed that C. pingtungensis was nested within Nipponcrysis. Our findings indicate that the taxonomy status of Candidia and Nipponcrysis is still needed to be confirmed.

Concepts: DNA, Gene, Biology, Organism, Species, Genetic code, Phylogenetics, Cyprinidae

5

The common carp, Cyprinus carpio, is one of the most important cyprinid species and globally accounts for 10% of freshwater aquaculture production. Here we present a draft genome of domesticated C. carpio (strain Songpu), whose current assembly contains 52,610 protein-coding genes and approximately 92.3% coverage of its paleotetraploidized genome (2n = 100). The latest round of whole-genome duplication has been estimated to have occurred approximately 8.2 million years ago. Genome resequencing of 33 representative individuals from worldwide populations demonstrates a single origin for C. carpio in 2 subspecies (C. carpio Haematopterus and C. carpio carpio). Integrative genomic and transcriptomic analyses were used to identify loci potentially associated with traits including scaling patterns and skin color. In combination with the high-resolution genetic map, the draft genome paves the way for better molecular studies and improved genome-assisted breeding of C. carpio and other closely related species.

Concepts: DNA, Gene, Genetics, Virus, Genome, Common carp, Carp, Cyprinidae

4

In many North American rivers, populations of multiple species of non-native cyprinid fishes are present, including black carp (), grass carp (), bighead carp ( nobilis), silver carp (), common carp (), and goldfish (). All six of these species are found in the Mississippi River basin and tracking their invasion has proven difficult, particularly where abundance is low. Knowledge of the location of the invasion front is valuable to natural resource managers because future ecological and economic damages can be most effectively prevented when populations are low. To test the accuracy of environmental DNA (eDNA) as an early indicator of species occurrence and relative abundance, we applied eDNA technology to the six non-native cyprinid species putatively present in a 2.6 river mile stretch of the Chicago (IL, USA) canal system that was subsequently treated with piscicide. The proportion of water samples yielding positive detections increased with relative abundance of the six species, as indicated by the number of carcasses recovered after poisoning. New markers for black carp, grass carp, and a common carp/goldfish are reported and details of the marker testing to ensure specificity are provided.

Concepts: River, Drainage basin, Carp, Cyprinidae, Goldfish, Mississippi River, Commercial fish, Bighead carp

3

The common carp (Cyprinus carpio) as one of the most important aquaculture fishes produces over 3 million metric tones annually, approximately 10% the annual production of the all farmed freshwater fish worldwide. However, the tetraploidy genome and long generation-time of the common carp have made its breeding and genetic studies extremely difficult. Here, TALEN and CRISPR-Cas9, two versatile genome-editing tools, are employed to target common carp bone-related genes sp7, runx2, bmp2a, spp1, opg, and muscle suppressor gene mstn. TALEN were shown to induce mutations in the target coding sites of sp7, runx2, spp1 and mstn. With CRISPR-Cas9, the two common carp sp7 genes, sp7a and sp7b, were mutated individually, all resulting in severe bone defects; while mstnba mutated fish have grown significantly more muscle cells. We also employed CRISPR-Cas9 to generate double mutant fish of sp7a;mstnba with high efficiencies in a single step. These results demonstrate that both TALEN and CRISPR-Cas9 are highly efficient tools for modifying the common carp genome, and open avenues for facilitating common carp genetic studies and breeding.

Concepts: DNA, Gene, Genetics, Fish, Carp, Cyprinidae, Goldfish, Aquaculture