Screening and activity of potential gastrointestinal probiotic lactic acid bacteria against Yersinia ruckeri O1b.

Yersiniosis of cultured Atlantic salmon is a recurrent fish health management challenge in many continents. The causative organism, Yersinia ruckeri, can reside latently in the gut and lead to acute infection and disease during hatchery and sea-transfer stages. One potential prevention approach is the administration of probiotic bacteria to suppress gut colonization of Y. ruckeri. Our study aimed to isolate and identify anti-Yersinia activity among lactic acid bacteria (LAB) isolated from the gastrointestinal tract (GIT) of aquatic animals. Of the 186 aquatic GIT isolates examined, three strains showed diffusible antimicrobial activity towards Y. ruckeri O1b. Analysis of 16 s rRNA gene sequences indicated the three bacterial strains were Enterococci, related to Enterococcus sp. (99%), Enterococcus thailandicus (99%), and Enterococcus durans (99%). Anti-Yersinia activity was maintained at neutral pH (~6.5-7.0), and in-vitro environmental tolerance assays showed the three strains could withstand simulated salmonids gastrointestinal tract conditions of: low pH (3.4) and 3% bile salt content. All three Enterococci strains showed higher adhesion to the intestinal mucus of Atlantic salmon than Y. ruckeri O1b (E. durans 24%, E. enterococcus sp. 25% and E. thailandicus 98%, compared to Y. ruckeri O1b 5%). However, only Enterococcus sp. and E. thailandicus were able to grow in the salmon intestinal mucus broth while E. durans showed no growth. Anti-Yersinia activity was completely inactivated by proteinase-K treatment, suggesting that the active compound/s are proteinaceous and may be bacteriocin-like inhibitory substances (BLIS). Our data indicate that Enterococcus sp. MA176 and E. thailandicus MA122 are potential probionts for the prevention of yersiniosis in salmonids. Further in-vivo studies are required to determine whether these bacteria reduce the incidence of yersiniosis in Atlantic salmon.

Prevalence, intensity and histopathology of Zeylanicobdella arugamensis infestation on groupers reared on different aquaculture systems.

The marine leech, Zeylanicobdella arugamensis, is a major threat to aquaculture in grouper-producing countries including Indonesia. This study aimed at investigating prevalence, intensity and histopathology of the ectoparasite in humpback and hybrid groupers cultured in different rearing systems. A total of 260 groupers (60 humpback groupers and 200 hybrid groupers) were used for samples. The marine leech was observed on skin, fins, gills and mouth, followed by histopathological assay on the skin tissue. The results showed that prevalence of the leech in both groupers was higher when they were cultured in the floating net cages compared with the hatchery, p < .05. Furthermore, humpback grouper had a higher prevalence than hybrid grouper when they were cultured in a similar system, p < .05. Meanwhile, there was no significant difference in intensity between the two groupers, p > .05. Within the hybrid groupers, the highest prevalence was obtained from hybrid groupers reared in the earthen ponds. Histopathological studies showed that the infected groupers exhibited inflammation, congestion and erosion of the epidermis layer. Hybrid grouper had more severe histopathological lesions in the skin tissues. These results suggested that species and type of aquaculture system had significantly determined the prevalence, intensity and severity of lesion in Z. arugamensis infestation.

Marine Organisms as Potential Sources of Bioactive Peptides that Inhibit the Activity of Angiotensin I-Converting Enzyme: A Review.

Angiotensin I-converting enzyme (ACE) is a paramount therapeutic target to treat hypertension. ACE inhibitory peptides derived from food protein sources are regarded as safer alternatives to synthetic antihypertensive drugs for treating hypertension. Recently, marine organisms have started being pursued as sources of potential ACE inhibitory peptides. Marine organisms such as fish, shellfish, seaweed, microalgae, molluscs, crustaceans, and cephalopods are rich sources of bioactive compounds because of their high-value metabolites with specific activities and promising health benefits. This review aims to summarize the studies on peptides from different marine organisms and focus on the potential ability of these peptides to inhibit ACE activity.

Refined linkage analysis of the sulphated marine polysaccharide fucoidan of Cladosiphon okamuranus with a focus on fucose.

Methylation followed by depolymerization and gas chromatography (GC) is an effective methodology for the linkage analysis of polysaccharides, including fucoidan, a sulphated algal polysaccharide. However, this sample material demands attention to experimental details to prevent aberrations in the analytical result. The use of deficient bases for methylation, the presence of water, analyte degradation during hydrolysis, and coelution of the target analytes during gas chromatography create doubts about published results. We therefore investigated critical parameters of the method and carefully optimized the steps of the protocol to ensure the integrity of the results for the fucose monomers. Fucoidan from Cladosiphon okamuranus was used as reference sample to determine the glycosidic bonds, and sulphate positions in the monomer. Fucoidan in protonated form was methylated in a strictly water-free environment using lithium dimsyl as base and methyl iodide for methylation. The methylated polymer was isolated by solid phase extraction, which was crucial to recover also the highly sulfated fraction. Hydrolysis was conducted with trifluoroacetic acid. To separate all target analytes in GC-FID/MS, a stationary phase with high cyanopropyl content (HP-88) was required, as the commonly employed phenyl siloxane phases result in co-elution, which distorts the result severely.

Acceleration of gonadal maturity of female striped catfish, Pangasianodon hypophthalmus (Pisces, Pangasiidae) by irradiation to low-power soft-laser.

This study was aimed to investigate the effects of red photodiode soft-laser irradiation on gonadal maturity and egg quality of female striped catfish, Pangasianodon hypophthalmus. Four female striped catfish (weighing 700-800 g) at stage I of gonadal maturation (for all treatments) were exposed four times (once a week) to different doses (0.2, 0.4, 0.6, and 0.8 J/cm2) of red photodiode soft-laser in the reproductive acupoint. Doses of 500 IU and 0.5 mL per kg body weight of human chorionic gonadotropin (hCG) and Ovaprim™, respectively were injected intramuscularly at week 4 (positive control), and no treatment (negative control). Finally, gonadal maturation stage (GMS), gonadosomatic index (GSI), hepatosomatic index (HSI), fecundity, and egg diameter were measured at week 5. The results showed that red photodiode soft-laser irradiation of 0.4 J/cm2 dose accelerated GMS IV at early week 3 (100 %) and increases GSI (3.42 %), HSI (1.26 %), fecundity (67,665 eggs/body weight), and egg diameter (1.01 mm) compared to the other treatments. Treatments of 0.2, 0.6, and 0.8 J/cm2 doses, and positive and negative controls only reached GMS III (75 %), III (25 %), I (100 %), II (25 %), and I (100 %), respectively. GSI values were 2.20 %, 1.47 %, 0.93 %, 0.44 %, and 0.42 %, respectively and HSI values were 1.07 %, 0.85 %, 0.7 7%, 0.66 %, and 0.53 %, respectively. Females irradiated with doses of 0.2 and 0.6 J/cm2 had fecundities of 38,822 and 29,530 eggs/body weight, respectively, and egg diameters of 0.88 and 0.29 mm, respectively. These results suggest that a red photodiode soft-laser irradiation accelerates gonadal maturity and improves egg quality in female striped catfish.

Prevalence of Antibiotic-Resistant Seafood-Borne Pathogens in Retail Seafood Sold in Malaysia: A Systematic Review and Meta-Analysis.

The objective of this study was to examine the frequency and extent of antibiotic-resistant pathogens in seafood sold in Malaysia, using a systematic review and meta-analysis approach to analyze primary research studies. Four bibliographic databases were systematically searched for primary studies on occurrence. Meta-analysis using a random-effect model was used to understand the prevalence of antibiotic-resistant bacteria in retail seafood sold in Malaysia. A total of 1938 primary studies were initially identified, among which 13 met the inclusion criteria. In the included primary studies, a total of 2281 seafoods were analyzed for the presence of antibiotic-resistant seafood-borne pathogens. It was observed that 51% (1168/2281) of the seafood was contaminated with pathogens. Overall, the prevalence of antibiotic-resistant seafood-borne pathogens in retail seafood was 55.7% (95% CI: 0.46-0.65). Antibiotic-resistant Salmonella species had an overall prevalence of 59.9% (95% CI: 0.32-0.82) in fish, Vibrio species had an overall prevalence of 67.2% (95% CI: 0.22-0.94) in cephalopods, and MRSA had an overall prevalence of 70.9% (95% CI: 0.36-0.92) in mollusks. It could be concluded that there is a high prevalence of antibiotic-resistant seafood-borne pathogens in the retail seafood sold in Malaysia, which could be of public health importance. Therefore, there is a need for proactive steps to be taken by all stakeholders to reduce the widespread transmission of antibiotic-resistant pathogens from seafood to humans.

Antioxidant, antimicrobial and tyrosinase inhibitory activities of xanthones isolated from Artocarpus obtusus F.M. Jarrett.

One of the most promising plants in biological screening test results of thirteen Artocarpus species was Artocarpus obtusus FM Jarrett and detailed phytochemical investigation of powdered dried bark of the plant has led to the isolation and identification of three xanthones; pyranocycloartobiloxanthone A (1), dihydroartoindonesianin C (2) and pyranocycloartobiloxanthone B (3). These compounds were screened for antioxidant, antimicrobial and tyrosinase inhibitory activities. Pyranocycloartobiloxanthone A (1) exhibited a strong free radical scavenger towards DPPH free radicals with IC50 value of 2 µg/mL with prominent discoloration observed in comparison with standard ascorbic acid, α-tocopherol and quercetin, The compound also exhibited antibacterial activity against methicillin resistant Staphylococcus aureus (ATCC3359) and Bacillus subtilis (clinically isolated) with inhibition zone of 20 and 12 mm, respectively. However the other two xanthones were found to be inactive. For the tyrosinase inhibitory activity, again compound (1) displayed strong activity comparable with the standard kojic acid.

Population Genetic Structure of Marine Leech, Pterobdella arugamensis in Indo-West Pacific Region.

Grouper aquaculture is rapidly expanding in both tropical and subtropical regions. The presence of marine leeches (Pterobdella arugamensis; previously named Zeylanicobdella arugamensis) infesting cultured groupers, however, can have a fatal effect on grouper aquaculture production and cause significant economic loss. Understanding the marine leech's population structure is therefore important to determine its possible distributional origin and distributional mechanisms, which will help monitor and mitigate the infestation. In this study, a total of 84 marine leeches collected from cultured hybrid groupers Epinephelus spp. in Brunei Darussalam, Malaysia and Indonesia were identified as P. arugamensis, based on morphological and mitochondrial cytochrome c oxidase subunit I gene sequence analyses. These leech samples, together with additional sequences from the GenBank database, were grouped into four genetically distinct haplogroups: (1) Asia Pacific, (2) Borneo, (3) Surabaya and (4) Iran. The four populations were found to be highly diverged from each other. The results also suggested that the samples from the Asia Pacific population could be dispersed and transported from Indonesia.

Nutritional quality and in vitro digestibility of fermented rice bran based on different types and doses of inoculants.

Objective: The study was conducted to determine the effect of inoculants of different types and doses on the nutrient quality and in vitro digestibility of fermented rice bran. Materials and Methods: The study was designed using a completely randomized design with a 3 × 3-factorial pattern. The first factor was the type of inoculum, consisting of Saccharomyces cerevisiae (SC), Effective Microorganism-4, and Saus Burger Pakan (SBP). The second factor is inoculum doses, which are as follows: levels 2%, 4%, and 6%. The variables measured included chemical composition, fiber fraction content, dry matter digestibility and organic matter digestibility. Results: The results showed that the type of inoculation treatment and the doses of inoculation did not affect the dry matter (DM) content of fermented bran, and the organic matter content of fermented bran was only affected by the inoculation dose factor (p < 0.05). The highest crude protein and Extract Ether (EE) were obtained in the SBP inoculants, which increased linearly with increasing inoculation doses (p < 0.05). While a significant decrease (p < 0.05) occurred in crude fiber content. The cellulose, hemicellulose, lignin, acid detergent fiber (ADF), and neutral detergent fiber (NDF) fractions were significantly lower in the SBP treatment as the dose increased. The SBP inoculant type produced the highest DMD (p < 0.05) but showed a response that was not different from the SC inoculant treatment for OMD. Increasing inoculation doses of 2%, 4%, and 6% linearly increased the DMD and OMD of fermented bran (p < 0.05). Overall, inoculant application on fermented bran showed an interaction effect except for the components of DM, EE, ADF, NDF, and DMD of fermented bran. Conclusions: It was concluded that the SBP at 6% and their combination resulted in the best chemical quality and digestibility of rice bran.

Topical Administration Effect of Sargassum duplicatum and Garcinia mangostana Extracts Combination on Open Wound Healing Process in Diabetic Mice.

This research aimed to determine the topical administration effect of the combination of Sargassum duplicatum and Garcinia mangostana extracts to ameliorate diabetic open wound healing. The study used 24 adult males of Mus musculus (BALB/c strain, 3-4 months, 30-40 g). They were divided into normal control groups (KN) and diabetic groups. The diabetic group was streptozotocin-induced and divided further into three treatment groups: the diabetic control group (KD), the S. duplicatum treatment group (PA), and the combination of S. duplicatum and G. mangostana treatment group (PAM). The dose of treatment was 50 mg/kg of body weight. Each group was divided into three treatment durations, which were 3 days, 7 days, and 14 days. The wound healing process was determined by wound width, the number of neutrophils, macrophages, fibroblasts, fibrocytes, and collagen density. Histological observation showed that the topical administration of combination extracts increased the re-epithelialization of the wounded area, fibroblasts, fibrocytes, and collagen synthesis. The topical administration of combination extracts also decreased the number of neutrophils and macrophages. This study concluded that the topical administration of the combination of S. duplicatum and G. mangostana extracts improved the open wound healing process in diabetic mice.

Identification of a novel umami peptide in tempeh (Indonesian fermented soybean) and its binding mechanism to the umami receptor T1R.

Tempeh, a traditional Indonesian soybean product produced by fermentation, is especially popular because of its umami taste. In this study, a novel umami peptide GENEEEDSGAIVTVK (GK-15) was identified in the small peptide (<3 kDa) fraction of the water extract of tempeh using LC-MS/MS analysis and database-assisted identification. The umami taste of GK-15 was further validated using sensory evaluation, which suggested that GK-15 may be one of the key components contributing to the umami taste in tempeh. To rationalize the biological effect of GK-15, molecular docking of GK-15 into the N-terminal extracellular ligand-binding domain of the umami (T1R) receptor was performed. ZDOCK data showed that GK-15 could perfectly bind either to the open or closed conformation of T1R3. To the best of our knowledge, the present work is the first study to focus on the screening of umami peptides from tempeh.

Detection of koi herpesvirus in healthy common carps, Cyprinus carpio L.

Koi herpesvirus (KHV), a member of Hervesviridae, has been frequently reported to cause mass mortality (80-100%) in common carps (Cyprinus carpio L.). A unique feature of Herverviridae members is latent infection, maintaining their genetic information for an extended period in the absence of productive infection, and reactivate when environmental conditions are favorable for their growth. To prevent this occurs, a monitoring program should be done for early detection. This study aimed at detecting the presence of KHV in healthy common carps reared in West-Nusa Tenggara Province, Indonesia. A total of 80 healthy fish was collected randomly from eight fish farms (Lingsar, Batu Kumbung, Narmada, Tanjung, Lenek, Aik Mel, Brang Rea and Rhee) across West-Nusa Tenggara Province, Indonesia. The presence of KHV genome was detected using a PCR with a commercial kit, IQ 2000TM. The result showed that common carps collected from four farms (Aik Mel, Lenek, Rhee and Brang Rea) were positive KHV. The size of an amplified gene was ~ 550 bp which was the same as positive KHV control. The obtained result suggests that KHV as other member of Hervesviridae shows a latent infection in common carps, and should be anticipated for their reactivation. Based on this result it is highly recommended that common carps cultured in this region should be vaccinated. In addition, transporting common carp out from Lombok and Sumbawa Islands should be carefully regulated to prevent the spread of the disease to other areas.