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Fulltext Identification and characterization of the Lactic Acid Bacteria isolated from Malaysian fermented fish (Pekasam)

Ida Muryany, M,Y., Ghazali, A.R., Hing, H.L., Nor Fadilah, R., Ina Salwany, M,Y.

International Food Research Journal, 2017;24(2):868-875.
MyJurnal

Recently researchers are interested with the biotherapeutic potential of probiotics in gut disease
treatment. The bacteria are generally regarded as a safe, have a stability of usage and originate
from the natural resources. The study aims to identify and characterize the potential probiotic
Lactic Acid Bacteria (LAB) isolated from Malaysian fermented fish product known as
Pekasam. Fourty isolates obtained were firstly screened for their antagonism activities against
the common pathogenic bacteria; Esherichia coli, Staphylococcus aureus and Klebsiella sp.
Our study revealed only three (labeled as L8, L20 and S1) of the isolates tested showed broad
antimicrobial effects towards the pathogenic bacteria. All of the isolates were also γ-hemolytic
and tolerant to various pH (pH 3, 5 and 7.5) and 0.3% (w/v) bile salts. The bacteria isolates
of strain L8 and L20 were susceptible to seven antibiotics tested except vancomycin and
tetracycline whereas S1 was resistant to all antibiotics. Phenotypic tests revealed that both
bacteria isolates of strain L8 and L20 were Bacillus megaterium while S1 was Pediococcus
pentosaceus whereas 16S rRNA gene sequence analysis showed potential bacteria isolates
of strain L8 and L20 belonged to the Lactobacillus plantarum (99% similarity) and S1 was
characterized as Lactobacillus pentosus (100% similarity) respectively. Our present study
showed that the probiotics of strain L8, L20 and S1 isolated from the fermented fish (Pekasam)
exhibited the potential probiotic properties to be developed as biotherapeutic agents.
Possible transmission routes of Vibrio spp. in tropical cage-cultured marine fishes

Nurliyana M, Amal MNA, Zamri-Saad M, Ina-Salwany MY

Lett Appl Microbiol, 2019 Jun;68(6):485-496.
PMID: 30834548 DOI: 10.1111/lam.13146

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.
Vaccination of Tilapia against Motile Aeromonas Septicemia: A Review

Shirajum Monir M, Yusoff SM, Mohamad A, Ina-Salwany MY

J Aquat Anim Health, 2020 06;32(2):65-76.
PMID: 32331001 DOI: 10.1002/aah.10099

The production of tilapia Oreochromis spp. is rapidly growing throughout the world, but atypical motile aeromonad septicemia (MAS) is a current threat to the tilapia farming industry. The etiological agent of this disease is usually Aeromonas hydrophila. Mortality rates due to MAS are frequently high, resulting in a devastating negative impact on this industry worldwide; therefore, proper control measures regarding both prevention and treatment are necessary. Although vaccines against MAS for tilapia are available, their effectiveness is entirely dependent on the specific strain of problematic bacteria. Until now, whole-cell inactivated A. hydrophila vaccines for tilapia have exhibited the highest level of protection over live attenuated and recombinant vaccines. Among the various vaccine administration systems, only intraperitoneal (i.p.) injections of the A. hydrophila vaccine into tilapia were found to provide prominent immune protection. Vaccine efficacy was primarily measured by using the i.p. injection challenge model and estimating the relative percent survival of the immunized tilapia. Freund's incomplete adjuvant showed to be the most effective for tilapia MAS vaccines. In this review, multiple factors that directly or indirectly influence the efficacy of MAS vaccines for tilapia (adjuvants, challenge models, immunization doses and duration, and size of vaccinated fish) are discussed.
Fulltext Isolation and partial characterization of Asian sea bass (Lates calcarifer) Vitellogenin

Fazielawanie NM, Siraj SS, Harmin SA, Ina-Salwany MY

Fish Physiol Biochem, 2013 Apr;39(2):191-200.
PMID: 22878544 DOI: 10.1007/s10695-012-9690-5

A study was conducted to isolate, partial characterize Asian sea bass (Lates calcarifer) vitellogenin (vtg). Two-year-old juvenile L. calcarifer (n = 10) were given three intraperitoneal injections of 17-β estradiol (E2) at a dose of 2 mg/kg body weight to induce vitellogenesis. Blood was collected 3 days after the last injection, and plasma was purified through gel filtration chromatography. A broad single symmetrical peak consisting of vtg molecule was produced. Protein concentration was 0.059 mg/ml as determined by Bradfrod assay using bovine serum albumin as a standard. The protein appeared as one circulating form in Native PAGE considering the dimeric form of putative vtg with molecular weight of 545 kDa. In SDS-PAGE under reducing conditions, two major bands appeared at 232.86 and 118.80 kDa and minor bands at 100.60, 85.80 and 39.92 kDa, respectively. The purified vtg was used to generate a polyclonal antibody, and the specificity of antibody was assessed by Western blot analysis. Two major bands were immunoreacted, but no cross-reactivity was observed with plasma from non-induced males. The protein was characterized as phosphoglycolipoprotein as it positively stained for the presence of lipid, phosphorus and carbohydrate using Sudan Black B, methyl green and periodic acid/Schiff reagent solution, respectively. The amino acid composition was analyzed by high sensitivity amino acid analysis that showed high percentage of non-polar amino acids (~48 %). The results suggest the potential utilization of vtg as a basis tool to further study about reproductive physiology of this important economical species.
Effects of skin abrasion in immersion challenge with Vibrio harveyi in Asian seabass Lates calcarifer fingerlings

Chin YK, Ina-Salwany MY, Zamri-Saad M, Amal MNA, Mohamad A, Lee JY, et al.

Dis Aquat Organ, 2020 Jan 16;137(3):167-173.
PMID: 31942862 DOI: 10.3354/dao03435

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.
Biofilm is associated with chronic streptococcal meningoencephalitis in fish

Isiaku AI, Sabri MY, Ina-Salwany MY, Hassan MD, Tanko PN, Bello MB

Microb Pathog, 2017 Jan;102:59-68.
PMID: 27890651 DOI: 10.1016/j.micpath.2016.10.029

Biofilms are aggregates of attached microbial organisms whose existence on tissues is often recognised as a mechanism for the establishment of most chronic diseases. Herein we investigated the ability of piscine Streptococcus agalactiae, an important aquatic pathogen, for adaptation to this sessile lifestyle in vitro and in the brain of a tilapia fish model. Piscine S. agalactiae exhibited a weak attachment to polystyrene plates and expressed a low biofilm phenotype under the study conditions. Furthermore, fluorescent in situ hybridization and confocal laser scanning microscopy revealed discrete aggregates of attached S. agalactiae within brain tissues and around meningeal surfaces. They were embedded in an exopolysaccharide containing matrix, intractable to inflammatory response and showed some level of resistance to penicillin despite proven susceptibility on sensitivity test. Intracellular bacterial aggregates were also observed, moreover, antibody mediated response was not demonstrated during infection. Nucleated erythrocytes appear to facilitate brain invasion possibly via the Trojan horse mechanism leading to a granulomatous inflammation. We have demonstrated that biofilm is associated with persistence of S. agalactiae and the development of chronic meningoencephalitis in fish.
Isolation and Pathogenicity of Streptococcus iniae in Cultured Red Hybrid Tilapia in Malaysia

Rahmatullah M, Ariff M, Kahieshesfandiari M, Daud HM, Zamri-Saad M, Sabri MY, et al.

J Aquat Anim Health, 2017 Dec;29(4):208-213.
PMID: 28787246 DOI: 10.1080/08997659.2017.1360411

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 β-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 feeding black soldier fly larvae on growth performance, protein, and fat content of red hybrid tilapia (Oreochromis spp.)

Aisyah HN, Athirah ZAR, Hanani WR, Arshad SS, Hassim HA, Nazarudin MF, et al.

Vet World, 2022 Oct;15(10):2453-2457.
PMID: 36425138 DOI: 10.14202/vetworld.2022.2453-2457

BACKGROUND AND AIM: In the aquaculture industry, the crucial goal is to minimize production costs, especially feeding costs, without significant side effects. Black soldier fly larva (BSFL) is a locally available, eco-friendly, and sustainable source that is high in crude protein (42% dry matter [DM]) and fat (35% DM). This study aimed to determine the growth performance along with the composition of crude fat and protein in red hybrid fingerlings after the addition of BSFL into the diet.
MATERIALS AND METHODS: A total of 120 fingerlings of uniform size (mean initial weight of 1.46 ± 0.06 g) were randomly assigned to one of four groups (n = 10) (A, B, C, and D) per tank (1 m × 2 m × 1 m). For 21 days, Group A (control group) was fed with 100% commercial diet; Group B was fed with 90% commercial fish diet + 10% BSFL; Group C was fed with 80% commercial fish diet + 20% BSFL; and Group D was fed with 70% commercial fish diet + 30% BSFL. Feed efficiency, growth performance, and proximate composition analysis were performed on the fish.
RESULTS: The results displayed that the group with the highest BSFL percentage had a greater effect on protein and fat composition than the control group. The proximate composition analysis of fish-fed diet revealed that an increase in the level of BSFL inclusion increases the protein content in the fish. In comparison to the other groups, the experimental diet with 30% BSFL inclusion has the highest levels of crude protein (80.30% DM) and fat (2.90% DM).
CONCLUSION: It is concluded that incorporating BSFL into a commercial diet for red hybrid tilapia fingerlings increased crude protein and fat composition, providing an alternative protein and fat source in fish diets.
Vibriosis in Fish: A Review on Disease Development and Prevention

Ina-Salwany MY, Al-Saari N, Mohamad A, Mursidi FA, Mohd-Aris A, Amal MNA, et al.

J Aquat Anim Health, 2019 03;31(1):3-22.
PMID: 30246889 DOI: 10.1002/aah.10045

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.