Effects of skin abrasion in immersion challenge with Vibrio harveyi in Asian seabass Lates calcarifer fingerlings.

Skin abrasions often occur in farmed fish following handling by labourers, injury by farm facilities, cannibalism and ectoparasites. Vibrio spp. are opportunistic pathogens that can invade host fish through damaged tissues and cause outbreaks of vibriosis. This study describes the effect of skin abrasions on the infectivity of V. harveyi using Asian seabass Lates calcarifer (Bloch, 1790) fingerlings as a case example and compares bacterial load and fish survival following immersion challenge with different doses. In total, 315 fish (6.67 ± 1.8 g) were divided into 3 treatments: skin abrasion followed by immersion infection, immersion infection only and an uninfected, uninjured control. Fish in the infection treatments were divided into 3 subgroups and exposed in triplicate to a 7 d immersion challenge with 106, 107 and 108 CFU ml-1 of live V. harveyi. No mortalities were observed in the control and immersion infection groups. However, fish in the skin abrasion treatment group that were infected with 108 CFU ml-1 of live V. harveyi showed signs of progressing disease throughout the experiment, which resulted in mortalities. Significantly higher bacterial loads (p < 0.05) were recorded in the intestine, liver and gills of the fish in this group. Fish in the skin abrasion treatment that were exposed to 107 and 108 CFU ml-1 of V. harveyi showed 100% mortality by Days 5 and 4, respectively. These findings confirm that skin injuries increase the susceptibility of seabass fingerlings to V. harveyi infection.

Possible transmission routes of Vibrio spp. in tropical cage-cultured marine fishes.

This study investigates the possible transmission routes of Vibrio spp. in a tropical cage-cultured marine fishes. Samplings of cultured Asian seabass, red snapper, hybrid grouper, wild fish, trash fish, fish fry, water and sediment samples were conducted from December 2016 to August 2017. All fish were dissected in situ and swabs were taken aseptically from the skin, eye, liver and kidney for bacterial isolation and identification. Bacterial isolation and identification from water, sediment and trash fish were also made. A total of 261 Vibrio spp. isolates recovered from the cultured, wild and fry fish, as well as from the sediment and water of the farm environment were analysed. Sequences of the pyrH gene were used to investigate the degree of relatedness and possible transmission routes existing between the isolated Vibrio spp. The population tree revealed the existence of selected Vibrio spp. that possibly transmitted between the newly introduced fish fry and wild fish into the cultured fish, while water also might possibly serves as natural transmission medium of certain Vibrio spp. in this fish farm. SIGNIFICANCE AND IMPACT OF THE STUDY: The source of transmission of Vibrio spp. into farmed marine fish remains unclear. This study highlights the possible transmission routes of Vibrio into cage-cultured marine fishes via newly introduced fish fry and wild fish. Understanding the routes of transmission of Vibrio spp. might help in controlling the disease in the near future.

Vibriosis in Fish: A Review on Disease Development and Prevention.

Current growth in aquaculture production is parallel with the increasing number of disease outbreaks, which negatively affect the production, profitability, and sustainability of the global aquaculture industry. Vibriosis is among the most common diseases leading to massive mortality of cultured shrimp, fish, and shellfish in Asia. High incidence of vibriosis can occur in hatchery and grow-out facilities, but juveniles are more susceptible to the disease. Various factors, particularly the source of fish, environmental factors (including water quality and farm management), and the virulence factors of Vibrio, influence the occurrence of the disease. Affected fish show weariness, with necrosis of skin and appendages, leading to body malformation, slow growth, internal organ liquefaction, blindness, muscle opacity, and mortality. A combination of control measures, particularly a disease-free source of fish, biosecurity of the farm, improved water quality, and other preventive measures (e.g., vaccination) might be able to control the infection. Although some control measures are expensive and less practical, vaccination is effective, relatively cheap, and easily implemented. In this review, the latest knowledge on the pathogenesis and control of vibriosis, including vaccination, is discussed.

The effects of fish gender on susceptibility to acute Streptococcus agalactiae infection in Javanese medaka Oryzias javanicus.

This study describes the susceptibility of different fish gender following acute Streptococcus agalactiae infection by using Javanese medaka Oryzias javanicus as test fish. The fish were grouped into four groups, which were: (1) all-male; (2) all-female; (3) mixed-gender (1 male: 1 female ratio); and (4) control non-infected (1 male: 1 female ratio). The fish in group 1, 2 and 3 were intraperitoneally exposed to 5.4 × 108 CFU/mL of S. agalactiae, while for group 4, the fish were exposed using sterile broth. The main clinical signs and histopathological changes of infected Javanese medaka were commonly observed in S. agalactiae infected fishes. However, no difference on clinical signs and histopathological changes of fish in group 1, 2 and 3 were noticed. The Javanese medaka mortality in group 1, 2 and 3 were observed from 4 h post infection (hpi) to 6 hpi, with the cumulative mortality from 3% to 30%. Then, the mortality increased at 12 hpi, with the range from 53% to 80%. However, 100% of the infected fish dead at 24 hpi. No clinical sign, histopathological change and fish mortality recorded in group 4. Generally, the clinical signs, mortality patterns, cumulative mortality and histopathological changes of Javanese medaka infected by S. agalactiae did not show any difference between the all-male, all-female and mixed-gender groups. This indicates that the susceptibility of fish to S. agalactiae infection is not influenced by their gender.

Isolation and Pathogenicity of Streptococcus iniae in Cultured Red Hybrid Tilapia in Malaysia.

This study describes the isolation and pathogenicity of Streptococcus iniae in cultured red hybrid tilapia (Nile Tilapia Oreochromis niloticus × Mozambique Tilapia O. mossambicus) in Malaysia. The isolated gram-positive S. iniae appeared punctiform, transparently white, catalase and oxidase negative and produced complete ß-hemolysis on blood agar, while a PCR assay resulted in the amplification of the 16 S rRNA gene and lactate oxidase encoded genes. The isolate was sensitive to tetracycline, vancomycin, and bacitracin but was resistant to streptomycin, ampicillin, penicillin, and erythromycin. Pathogenicity trials conducted in local red hybrid tilapia (mean ± SE = 20.00 ± 0.45 g) showed 90.0, 96.7, and 100.0% mortality within 14 d postinfection following intraperitoneal exposure to 104, 106, and 108 CFU/mL of the pathogen, respectively. The clinical signs included erratic swimming, lethargy, and inappetance at 6 h postinfection, while mortality was recorded at less than 24 h postinfection in all infected groups. The LD50-336 h of S. iniae against the red hybrid tilapia was 102 CFU/mL. The post mortem examinations revealed congested livers, kidneys, and spleens of the infected fish. This is the first report of S. iniae experimental infection in cultured red hybrid tilapia in Malaysia. Received January 20, 2017; accepted July 16, 2017.

The effect of feed-based vaccination on tilapia farm endemic for streptococcosis.

A tilapia farm experiencing endemic streptococcosis was selected to study the effect of vaccination with a feed-based vaccine on naturally ocurring streptococcosis. A total of 9000 red tilapia, Oreochromis niloticus × Oreochromis mossambicus of 100 ± 20 g were divided into 9 cages. Fish of Group 1 in cages 1, 2 and 3 were not vaccinated. Group 2 in cages 4, 5 and 6 were vaccinated on days 0 and 14 (single booster) while Group 3 in cages 7, 8 and 9 were vaccinated on days 0, 14 and 42 (double booster). Vaccination was done by oral administration of the feed-based bacterin vaccine at 4% bodyweight. Samples of serum for antibody study and the brain, eyes and kidney for bacterial isolation were collected at 14-day intervals. The study was carried out during the critical months between April and June. Following vaccination and booster, there was significant (p < 0.05) increase in the antibody levels in all vaccinated groups from week 1 that reached the peak at week 3 before declining gradually until week 6. However, second booster on week 6 significantly (p < 0.05) increased the antibody level that remained high until the end of the 16-week study period (double booster). Streptococcus agalactiae was isolated at the start of the experiment (day 0) at an average of 10 ± 5.0% of the sampled fish. In week 4, the isolation rate was 13 ± 5.7% but increased to 18 ± 7.6% in week 8, to 25 ± 10.0% in week 10, to 28 ± 5.8% in week 12 and 25 ± 7.3% in week 14. The average isolation rate was 28 ± 7.2%, 18 ± 7.1% and 13 ± 8.2% of the fish sampled from unvaccinated, single booster and double booster groups, respectively. At the end of the study period, the survival rate was 45.2 ± 2.45% for unvaccinated, 65.3 ± 4.8% for single booster and 75.1 ± 2.1% for double booster groups. Vaccinating fish in endemic farm might not eliminate the disease but was able to significantly improve the survival rate.

Feed-based vaccination regime against streptococcosis in red tilapia, Oreochromis niloticus x Oreochromis mossambicus.

BACKGROUND: Streptococcosis is an important disease of tilapia throughout the world. In Malaysia, streptococcosis outbreak was commonly reported during the 3-month period of high water temperature between April and July. This study describes the duration of protection following single and double booster dose regimes against streptococcosis in tilapia using a feed-based vaccine containing formalin-killed Streptococcus agalactiae. A total of 510 tilapias of 120 ± 10 g were selected and divided into 3 groups. Fish of Group 1 were vaccinated at weeks 0 and 2 (single booster group) while fish of Group 2 were vaccinated at weeks 0, 2 and 6 (double booster group) with a feed-based vaccine against streptococcosis. Fish of Group 3 was not vaccinated. Serum samples were collected weekly to determine the antibody level while samples of eye, brain and kidney were collected for bacterial isolation. At week 10, all fish were challenged with live S. agalactiae and the survival rate was determined. RESULTS: Both vaccinated groups showed significant (p < 0.05) increase in the antibody levels following the first booster dose, which lasted until week 6. Group 2 showed consistent high level of antibody following the second booster dose at week 6 and remained high until week 12. Challenge trial at week 10 resulted in 45 %, 70 % and 0 % rate of survival for Groups 1, 2 and 3, respectively. CONCLUSION: Double booster regime is most suitable to be applied for feed-based vaccination against streptococcosis prior to the start of the hot season.

Molecular characterization of Streptococcus agalactiae strains isolated from fishes in Malaysia.

AIMS: The aim of this study was to characterize Streptococcus agalactiae strains that were isolated from fishes in Malaysia using random amplified polymorphic DNA (RAPD) and repetitive extragenic palindromic PCR (REP-PCR) techniques. METHODS AND RESULTS: A total of 181 strains of Strep. agalactiae isolated from red hybrid tilapia (Oreochromis sp.) and golden pompano (Trachinotus blochii) were characterized using RAPD and REP-PCR techniques. Both the fingerprinting techniques generated reproducible band patterns, differing in the number and molecular mass amplicons. The RAPD technique displayed greater discriminatory power by its production of more complex binding pattern and divided all the strains into 13 groups, compared to 9 by REP-PCR technique. Both techniques showed the availability to differentiate the genetic profiles of the strains according to their geographical location of origin. Three strains of Strep. agalactiae that were recovered from golden pompano showed a genetic dissimilarity from the strains isolated from red hybrid tilapia, while the strain of ATCC 27956 that recovered from bovine displayed a unique profile for both methods. CONCLUSIONS: Both techniques possess excellent discriminative capabilities and can be used as a rapid means of comparing Strep. agalactiae strains for future epidemiological investigation. SIGNIFICANCE AND IMPACT OF THE STUDY: Framework as the guideline in traceability of this disease and in the search for potential local vaccine candidates for streptococcosis in this country.

Pathological changes in red tilapias (Oreochromis spp.) naturally infected by Streptococcus agalactiae.

The pathological changes present in 300 red tilapias (Oreochromis spp.) naturally infected by Streptococcus agalactiae are described. The most consistent gross findings were marked congestion of internal organs, particularly the liver, spleen and kidneys. Other features included exophthalmos, softening of the brain and the occasional accumulation of fluid within the abdominal cavity. Microscopical examination confirmed the presence of marked congestion of the liver, spleen and kidneys. The endothelial cells lining major blood vessels of the liver and occasionally the spleen were swollen and vacuolated. There was evidence of vascular thrombosis with infarction of surrounding tissue. Bacterial colonies were noted within and immediately surrounding the affected blood vessels. The meninges were thickened by the infiltration of numerous heterophils. Similar infiltrates of heterophils and lymphocytes were observed in the lamina propria of the intestine. The kidneys were severely congested and haemorrhagic, with extensive interstitial nephritis.