ABSTRACT
As one of the most important aquatic fish, Micropterus salmoides suffers lethal and epidemic disease caused by rhabdovirus at the juvenile stage. In this study, a new strain of M. salmoides rhabdovirus (MSRV) was isolated from Yuhang, Zhejiang Province, China, and named MSRV-YH01. The virus infected the grass carp ovary (GCO) cell line and displayed virion particles with atypical bullet shape, 300-500 nm in length and 100-200 nm in diameter under transmission electron microscopy. The complete genome sequence of this isolate was determined to include 11 526 nucleotides and to encode five classical structural proteins. The construction of the phylogenetic tree indicated that this new isolate is clustered into the Vesiculovirus genus and most closely related to the Siniperca chuatsi rhabdovirus. To explore the potential for a vaccine against MSRV, a glycoprotein (1-458 amino acid residues) of MSRV-YH01 was successfully amplified and cloned into the plasmid pFastBac1. The high-purity recombinant bacmid-glycoprotein was obtained from DH10Bac through screening and identification. Based on polymerase chain reaction (PCR), western blot, and immunofluorescence assay, recombinant virus, including the MSRV-YH01 glycoprotein gene, was produced by transfection of SF9 cells using the pFastBac1-gE2, and then repeatedly amplified to express the glycoprotein protein. We anticipate that this recombinant bacmid system could be used to challenge the silkworm and develop a corresponding oral vaccine for fish.
Subject(s)
Animals , Female , Baculoviridae/metabolism , Bass/metabolism , Carps/virology , Cell Line , Genetic Techniques , Genome, Viral , Glycoproteins/biosynthesis , Insecta , Ovary/virology , Phylogeny , Plasmids/metabolism , Recombinant Proteins/biosynthesis , Rhabdoviridae/metabolismABSTRACT
As one of the most important aquatic fish, Micropterus salmoides suffers lethal and epidemic disease caused by rhabdovirus at the juvenile stage. In this study, a new strain of M. salmoides rhabdovirus (MSRV) was isolated from Yuhang, Zhejiang Province, China, and named MSRV-YH01. The virus infected the grass carp ovary (GCO) cell line and displayed virion particles with atypical bullet shape, 300–500 nm in length and 100–200 nm in diameter under transmission electron microscopy. The complete genome sequence of this isolate was determined to include 11 526 nucleotides and to encode five classical structural proteins. The construction of the phylogenetic tree indicated that this new isolate is clustered into the Vesiculovirus genus and most closely related to the Siniperca chuatsi rhabdovirus. To explore the potential for a vaccine against MSRV, a glycoprotein (1–458 amino acid residues) of MSRV-YH01 was successfully amplified and cloned into the plasmid pFastBac1. The high-purity recombinant bacmid-glycoprotein was obtained from DH10Bac through screening and identification. Based on polymerase chain reaction (PCR), western blot, and immunofluorescence assay, recombinant virus, including the MSRV-YH01 glycoprotein gene, was produced by transfection of SF9 cells using the pFastBac1-gE2, and then repeatedly amplified to express the glycoprotein protein. We anticipate that this recombinant bacmid system could be used to challenge the silkworm and develop a corresponding oral vaccine for fish.
ABSTRACT
Present study was performed to survey infection status of digenetic trematode metacercariae in 2 alien fish species, Micropterus salmoides (largemouth bass) and Lepomis macrochirus (bluegill), in 2 rivers draining Chungcheongbuk-do, Republic of Korea. A total of 107 largemouth bass and 244 bluegills were caught in Daecheong-ho (ho=lake) and Musim-cheon (a branch of Geum-gang), in Chungcheongbuk-do April–July 2015. Additionally, 68 native fish of 5 species, i.e., Zacco platypus, Hemibarbus longirostris, Carassius auratus, Pseudogobio esocinus and Puntungia herzi, were caught from the same water bodies. All of the fish collected were examined by artificial digestion method. The metacercariae of Centrocestus armatus, Clinostomum complanatum, Metagonimus sp. and Diplostomum spp. were detected from 4 out of 5 native fish species in Daecheong-ho. However, any metacercariae were not found from 87 M. salmoides and 177 L. macrochirus in Daecheong-ho. In Musim-cheon, metacercariae of Exorchis oviformis and Metacercaria hasegawai were detected from 78% Z. platypus and 34% L. macrochirus, but any metacercariae not found in M. salmoides. We report here that the 2 alien fish species were less infected with the metacercariae than the native ones. Surveys on the metacercariae in the alien fish species in geographically various rivers should be undertaken for better understanding on the role of alien fish species in the trematode infections in Republic of Korea.
Subject(s)
Humans , Bass , Digestion , Emigrants and Immigrants , Goldfish , Heterophyidae , Metacercariae , Methods , Perciformes , Platypus , Republic of Korea , Rivers , Trematode Infections , WaterABSTRACT
Micropterus salmoides is an important fish species for sport fishing activities, condition that has promoted its introduction to different reservoirs in Mexico and worldwide. With the aim to improve its fisheries management, this research dealt with some reproductive aspects of this species in the Gustavo Díaz Ordaz reservoir, where it was studied from August 2008 through March 2011. To this end, we obtained 938 specimens, with gillnets of different sizes, to determine their total length (Lt, in cm), weight (Pt, in g), sex, gonadosomatic index, condition factor, fecundity and size at first maturity. Lt and Pt ranged from 15.9 to 63cm (37.4±78.0) and 57 to 4 431g (731.7±619.0), respectively. The Pt-Lt relationship showed a positive allometric growth, with no significant difference between males and females (F=0.9955, p=0.3187). The male: female ratio obtained was 1:0.83. Mass spawning lasted from December to April. Size at first maturity was 33.7cm and average fecundity was 32 294±12 878.7 oocytes/female. The gonadosomatic index was low from May through November, and increased between January and March. The condition factor was high before the spawning season and decreased after the reproductive period. We recommend a fishing ban from January to March, and to allow the capture size between 33 and 40cm.
Micropterus salmoides es un pez de importancia para la pesca deportiva en la mayoría de los embalses de México y del mundo, por lo que en el embalse Gustavo Díaz Ordaz se realizó un estudio sobre aspectos reproductivos entre agosto 2008 y marzo 2011. Se recolectaron 938 organismos, a los cuales se les midió la longitud (Lt cm), peso (Pt g), se determinó el sexo, índice gonadosomático, factor de condición, fecundidad y talla de primera madurez. La longitud y peso de los ejemplares recolectados varió de 15.9 a 63 cm (37.4±78.0) y de 57 a 4 431g (731.7±619.0), respectivamente. La relación peso total-longitud total se ajustó a un modelo tipo potencial, el crecimiento fue alométrico positivo, no se obtuvieron diferencias significativas entre machos y hembras (F=0.9955, p=0.3187). La proporción macho-hembra fue 1:0.83. Los desoves masivos inician en diciembre y finalizan en abril. La talla de primera reproducción fue de 33.7cm y la fecundidad promedio de 32 294±12 878.7 ovocitos/hembra. El índice gonadosomático se mantuvo bajo desde mayo hasta noviembre y alto entre enero y marzo. El factor de condición registró valores altos previos al desove y disminuyó al final del periodo reproductivo.