scholarly journals Identification and Genome Analysis of Vibrio coralliilyticus Causing Mortality of Pacific Oyster (Crassostrea gigas) Larvae

Pathogens ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 206 ◽  
Author(s):  
Hyoun Joong Kim ◽  
Jin Woo Jun ◽  
Sib Sankar Giri ◽  
Cheng Chi ◽  
Saekil Yun ◽  
...  

Vibrio coralliilyticus is known as a coral pathogen that also infects marine bivalve larvae worldwide. It is considered to be one of the major constraints in artificial marine bivalve seed production as it causes mortality. In this study, we first isolated and characterized a high virulent of V. coralliilyticus designated as SNUTY-1 that was the cause of Pacific oyster larvae mortality in Korea. In the pathogenicity test, exposure to 2.14 × 105 CFU/mL for 24 h caused mortality to 88.65 ± 2.4% of the tested healthy Pacific oyster larvae. SNUTY-1 showed anti-microbial resistance to β-lactams, such as penicillins, cephalosporins, and carbapenems. We sequenced and assembled the complete genome of SNUTY-1 (5,842,676 bp), consisting of two chromosomes (Chr I and Chr II) and two plasmids (pSNUTY1 and pSNUTY2). The COG functional analysis confirmed that Chr I had more genes associated with basic cellular functions in comparison to Chr II. The results of the phylogenetic trees based on OrthoANI values indicated that the SNUTY-1 was closely related to V. coralliilyticus strains. SNUTY-1 had a unique plasmid (pSNUTY2), which could mean that the Korean isolate is different from other sequenced V. coralliilyticus strains from different geographical origins. Toxic proteins such as cytolysin/hemolysin and extracellular metalloprotease genes were encoded on Chr I and Chr II of SNUTY-1. These data facilitate the control of V. coralliilyticus infections in aquaculture by providing valuable insights into the biodiversity of this organism and valuable information for the study of virulence factors.

2018 ◽  
Vol 6 (2) ◽  
Author(s):  
Hyoun Joong Kim ◽  
Jin Woo Jun ◽  
Sib Sankar Giri ◽  
Cheng Chi ◽  
Saekil Yun ◽  
...  

ABSTRACT We report here the complete genome sequence of the Vibrio coralliilyticus-specific phage pVco-5, a double-stranded DNA virus isolated from an oyster hatchery tank. Vibrio coralliilyticus causes bacillary necrosis in marine bivalve larvae; hence, phage pVco-5 could be used to prevent V. coralliilyticus infections in these larvae.


Pathogens ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 831
Author(s):  
Hyoun Joong Kim ◽  
Jin Woo Jun ◽  
Sib Sankar Giri ◽  
Sang Guen Kim ◽  
Sang Wha Kim ◽  
...  

Vibrio coralliilyticus (V. coralliilyticus) is a pathogen that causes mass mortality in marine bivalve hatcheries worldwide. In this study, we used a bacteriophage (phage) cocktail to prevent multiple-antibiotic-resistant (MAR) and phage-resistant (PR) V. coralliilyticus infection in Pacific oyster (Crassostreagigas) larvae. To prevent the occurrence of phage-resistant strains and decrease the effect of mono-phage treatment, we prepared a phage cocktail containing three types of V. coralliilyticus-specific phages and tested its prophylactic efficacy against MAR and PR V. coralliilyticus infection. The results of the cell lysis test showed that the phage cocktail showed an excellent bactericidal effect against the MAR and PR variants in contrast to the experimental group treated with two mono phages (pVco-5 and pVco-7). An in vivo test using Pacific oyster larvae also confirmed the preventive effect against MAR and PR variants. In conclusion, the application of the phage cocktail effectively prevented V. coralliilyticus infection in marine bivalve seedling production. Furthermore, it is expected to reduce damage to the aquaculture industry caused by the occurrence of MAR and PR V. coralliilyticus. Therefore, phage cocktails may be used for the control of various bacterial diseases.


2020 ◽  
Vol 8 (6) ◽  
pp. 926 ◽  
Author(s):  
Hyoun Joong Kim ◽  
Sib Sankar Giri ◽  
Sang Guen Kim ◽  
Sang Wha Kim ◽  
Jun Kwon ◽  
...  

Vibrio coralliilyticus is one of the major pathogens causing mass mortality in marine bivalve larvae aquaculture. To prevent and control Vibrio spp. infections in marine bivalve hatcheries, various antibiotics are overused, resulting in environmental pollution and the creation of multi-drug-resistant strains. Therefore, research on the development of antibiotic substitutes is required. In this study, we isolated two bacteriophages (phages) that specifically infected pathogenic V. coralliilyticus from an oyster hatchery and designated them as pVco-5 and pVco-7. Both phages were classified as Podoviridae and were stable over a wide range of temperatures (4–37 °C) and at pH 7.0–9.0. Thus, both phages were suitable for application under the environmental conditions of an oyster hatchery. The two phages showed confirmed significant bactericidal efficacy against pathogenic V. coralliilyticus in an in vitro test. In the in vivo experiment, the phage pre-treated groups of Pacific oyster larvae showed significantly lower mortality against V. coralliilyticus infection than untreated control larvae. The results of the present study suggest that both phages could be used in the artificial marine bivalve seedling industry; not only to prevent pathogenic V. coralliilyticus infection, but also to reduce antibiotic overuse.


Agronomy ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1955
Author(s):  
Anysia Hedy Ujat ◽  
Ganesan Vadamalai ◽  
Yukako Hattori ◽  
Chiharu Nakashima ◽  
Clement Kiing Fook Wong ◽  
...  

The re-emergence of the Fusarium wilt caused by Fusarium odoratissimum (F. odoratissimum) causes global banana production loss. Thirty-eight isolates of Fusarium species (Fusarium spp.) were examined for morphological characteristics on different media, showing the typical Fusarium spp. The phylogenetic trees of Fusarium isolates were generated using the sequences of histone gene (H3) and translation elongation factor gene (TEF-1α). Specific primers were used to confirm the presence of F. odoratissimum. The phylogenetic trees showed the rich diversity of the genus Fusarium related to Fusarium wilt, which consists of F. odoratissimum, Fusarium grosmichelii, Fusarium sacchari, and an unknown species of the Fusarium oxysporum species complex. By using Foc-TR4 specific primers, 27 isolates were confirmed as F. odoratissimum. A pathogenicity test was conducted for 30 days on five different local cultivars including, Musa acuminata (AAA, AA) and Musa paradisiaca (AAB, ABB). Although foliar symptoms showed different severity of those disease progression, vascular symptoms of the inoculated plantlet showed that infection was uniformly severe. Therefore, it can be concluded that the Fusarium oxysporum species complex related to Fusarium wilt of banana in Malaysia is rich in diversity, and F. odoratissimum has pathogenicity to local banana cultivars in Malaysia regardless of the genotype of the banana plants.


Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1278-1278 ◽  
Author(s):  
S. E. Cho ◽  
J. H. Park ◽  
S. H. Hong ◽  
I. Y. Choi ◽  
H. D. Shin

Agastache rugosa (Fisch. & C.A. Mey.) Kuntze, known as Korean mint, is an aromatic plant in the Lamiaceae. It is widely distributed in East Asian countries and is used as a Chinese traditional medicine. In Korea, fresh leaves are commonly added to fish soups and stews (3). In November 2008, several dozen Korean mints plants growing outdoors in Gimhae City, Korea, were found to be severely infected with a powdery mildew. The same symptoms had been observed in Korean mint plots in Busan and Miryang cities from 2008 to 2013. Symptoms first appeared as thin white colonies, which subsequently developed into abundant hyphal growth on stems and both sides of the leaves. Severe disease pressure caused withering and senescence of the leaves. Voucher specimens (n = 5) were deposited in the Korea University Herbarium (KUS). Appressoria on the mycelium were nipple-shaped or nearly absent. Conidiophores were 105 to 188 × 10 to 13 μm and produced 2 to 4 immature conidia in chains with a sinuate outline, followed by 2 to 3 cells. Foot-cells of the conidiophores were straight, cylindrical, slightly constricted at the base, and 37 to 58 μm long. Conidia were hyaline, ellipsoid to barrel-shaped, measured 25 to 40 × 15 to 23 μm (length/width ratio = 1.4 to 2.1), lacked distinct fibrosin bodies, and showed reticulate wrinkling of the outer walls. Primary conidia were obconically rounded at the apex and subtruncate at the base. Germ tubes were produced at the perihilar position of conidia. No chasmothecia were observed. The structures described above were typical of the Oidium subgenus Reticuloidium anamorph of the genus Golovinomyces. The measurements and morphological characteristics were compatible with those of G. biocellatus (Ehrenb.) V.P. Heluta (1). To confirm the identification, molecular analysis of the sequence of the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) of isolate KUS-F27200 was conducted. The complete ITS rDNA sequence was amplified using primers ITS5 and P3 (4). The resulting 514-bp sequence was deposited in GenBank (Accession No. KJ585415). A GenBank BLAST search of the Korean isolate sequence showed >99% similarity with the ITS sequence of many G. biocellatus isolates on plants in the Lamiaceae (e.g., Accession Nos. AB307669, AB769437, and JQ340358). Pathogenicity was confirmed by gently pressing diseased leaf onto leaves of five healthy, potted Korean mint plants. Five non-inoculated plants served as a control treatment. Inoculated plants developed symptoms after 7 days, whereas the control plants remained symptomless. The fungus present on inoculated plants was identical morphologically to that observed on the original diseased plants. The pathogenicity test was repeated with identical results. A powdery mildew on A. rugosa caused by G. biocellatus was reported from Romania (2). To our knowledge, this is the first report of powdery mildew caused by G. biocellatus on A. rugosa in Korea. The plant is mostly grown using organic farming methods with limited chemical control options. Therefore, alternative control measures should be considered. References: (1) U. Braun and R. T. A. Cook. Taxonomic Manual of the Erysiphales (Powdery Mildews), CBS Biodiversity Series No. 11. CBS, Utrecht, 2012. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Syst. Mycol. Microbiol. Lab., online publication, USDA ARS, retrieved 17 February 2014. (3) T. H. Kim et al. J. Sci. Food Agric. 81:569, 2001. (4) S. Takamatsu et al. Mycol. Res. 113:117, 2009.


2018 ◽  
Vol 9 ◽  
Author(s):  
Loïc Jacquemot ◽  
Yvan Bettarel ◽  
Joanne Monjol ◽  
Erwan Corre ◽  
Sébastien Halary ◽  
...  

2019 ◽  
Vol 167 ◽  
pp. 107244 ◽  
Author(s):  
Hyoun Joong Kim ◽  
Jin Woo Jun ◽  
Sib Sankar Giri ◽  
Cheng Chi ◽  
Saekil Yun ◽  
...  

2011 ◽  
Vol 91 (7) ◽  
pp. 1567-1576 ◽  
Author(s):  
S.A. Heaney ◽  
A.P. Maloy ◽  
J.W. Slater

Improved understanding of the occurrence and spatio-temporal distribution of bivalve larvae holds significant benefits for ecological studies, shellfisheries management and aquaculture. Morphological methods for identification have proved difficult to develop because of the small size of these larvae and similarities in their shape and colour. Molecular methods based on DNA extraction can confirm the presence of a species in a plankton sample, but without sample sorting and individual larval analysis, provide no estimate of larval abundance and are incapable of providing an estimate of larval growth rate. Fluorescencein situhybridization (FISH) using species-specific DNA probes has the potential to resolve these issues. However, utilization of this technique is constrained by the strong autofluorescence, common in marine larvae. Here we evaluate the effect of eight different fixatives on the autofluorescence intensity of bivalve larvae using fluorescein isothiocyanate (FITC) and Cy3 filters. In addition, fifteen autofluorescence reduction treatments were evaluated and their compatibility with FISH assessed. Relative to fresh larvae, chemically fixed larvae had significantly higher autofluorescence in both filter sets. Larvae preserved by freezing at –80°C exhibited no significant increase in autofluorescence over a 3-year period. Autofluorescence levels were generally lower with the FITC filter set than the Cy3 filter set. For archived larvae preserved in modified saline ethanol and exhibiting fixative-induced autofluorescence, the autofluorescence intensity could be reduced to 20–30% with saturated Sudan Black B and to 30–40% with Chemicon™. Both of these autofluorescence reduction treatments were compatible with subsequent FISH protocols using a FITC-labelled probe.


Reproduction ◽  
2014 ◽  
Vol 147 (3) ◽  
pp. 331-345 ◽  
Author(s):  
Sayyed Mohammad Hadi Alavi ◽  
Natsuki Matsumura ◽  
Kogiku Shiba ◽  
Naoki Itoh ◽  
Keisuke G Takahashi ◽  
...  

Factors that inhibit and stimulate the initiation of sperm motility were determined for Manila clam (Ruditapesphilippinarum), Pacific oyster (Crassostrea gigas), and Japanese scallop (Patinopecten yessoensis). Compared with artificial seawater (ASW), serotonin (5-hydroxytryptamine creatinine sulfate, 5-HT) could fully trigger sperm motility and increase sperm velocity and motility duration. Sperm motility was decreased in ASW at pH 6.5–7.0 and suppressed at pH 4.0. In Manila clam and Pacific oyster, 5-HT could overcome the inhibitory effects of acidic pH on sperm motility. In the presence of nigericin (a K+/H+exchanger), sperm motility was only triggered at pH 8.3. Testicular fluid K+concentrations were two- to fourfold higher than that in ASW. Sperm motility and velocity were decreased in ASW or 5-HT containing ≥40 mM K+or ≥2.5 mM 4-aminopyridine, suggesting K+efflux requirement to initiate motility. Sperm motility and velocity were reduced in ASW or 5-HT containing EGTA or W-7, suggesting that extracellular Ca2+is required for Ca2+/calmodulin-dependent flagellar beating. Ca2+influx occurs via Ca2+channels because sperm motility and velocity were decreased in both ASW and 5-HT containing T-type and L-type Ca2+channel blockers. 5-HT-dependent initiation of sperm motility was associated with intracellular Ca2+rise, which was comparable to that seen in ASW but was not observed in the presence of EGTA or a Ca2+channel blocker. Extracellular Na+is also essential for sperm motility initiation via regulation of Na+/Ca2+exchange. Overall, 5-HT-dependent initiation of sperm motility in marine bivalve mollusks is an osmolality-independent mechanism and regulated by extracellular pH, K+, Ca2+, and Na+.


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