Assessment of fish protein hydrolysate as a substitute for fish meal in white shrimp diets: Impact on growth, immune response, and resistance against Vibrio parahaemolyticus infection.

This study investigated the effects of fish protein hydrolysate derived from barramundi on growth performance, muscle composition, immune response, disease resistance, histology and gene expression in white shrimp (Penaeus vannamei). In vitro studies demonstrated FPH enhanced mRNA expressions of key immune-related genes and stimulated reactive oxygen species (ROS) production and phagocytic activity in shrimp hemocytes. To evaluate the effects of substituting fish meal with FPH in vivo, four isoproteic (43 %), isolipidic (6 %), and isoenergetic diets (489 kcal/100 g) were formulated with fish meal substitution levels of 0 % (control), 30 % (FPH30), 65 % (FPH65), and 100 % (FPH100). After 8-week feeding, the growth performance of FPH65 and FPH100 were significantly lower than that of control and FPH30 (p < 0.05). Similarly, the midgut histological examination revealed the wall thickness and villi height of FPH100 were significantly lower than those of control (p < 0.05). The shrimps were received the challenge of AHPND + Vibrio parahaemolyticus at week 4 and 8. All FPH-fed groups significantly enhanced resistance against Vibrio parahaemolyticus at week 4 (p < 0.05). However, this protective effect diminished after long-period feeding. No significant difference of survival rate was observed among all groups at week 8 (p > 0.05). The expressions of immune-related genes were analyzed at week 4 before and after challenge. In control group, V. parahaemolyticus significantly elevated SOD in hepatopancreas and Muc 19, trypsin, Midline-fas, and GPx in foregut (p < 0.05). Moreover, hepatopancreatic SOD of FPH65 and FPH100 were significantly higher than that of control before challenge (p < 0.05). Immune parameters were measured at week 8. Compared with control, the phagocytic index of FPH 30 was significantly higher (p < 0.05). However, dietary FPH did not alter ROS production, phenoloxidase activity, phagocytic rate, and total hemocyte count (p > 0.05). These findings suggest that FPH30 holds promise as a feed without adverse impacts on growth performance while enhancing the immunological response of white shrimp.

Improved accuracy in IoT-Based water quality monitoring for aquaculture tanks using low-cost sensors: Asian seabass fish farming.

Traditional approaches to monitoring water quality in aquaculture tanks present numerous limitations, including the inability to provide real-time data, which can lead to improper feeding practices, reduced productivity, and potential environmental risks. To address these challenges, this study aimed to create an accurate water quality monitoring system for Asian seabass fish farming in aquaculture tanks. This was achieved by enhancing the accuracy of low-cost sensors using simple linear regression and validating the IoT system data with YSI Professional Pro. The system's development and validation were conducted over three months, employing professional devices for accuracy assessment. The accuracy of low-cost sensors was significantly improved through simple linear regression. The results demonstrated impressive accuracy levels ranging from 76% to 97%. The relative error values which range from 0.27% to 4% demonstrate a smaller range compared to the values obtained from the YSI probe during the validation process, signifying the enhanced accuracy and reliability of the IoT sensor by using simple linear regression. The system's enhanced accuracy facilitates convenient and reliable real-time water quality monitoring for aquafarmers. Real-time data visualization was achieved through a microcontroller, Thingspeak, Virtuino application, and ESP 8266 Wi-Fi module, providing comprehensive insights into water quality conditions. Overall, this adaptable tool holds promise for accurate water quality management in diverse aquatic farming practices, ultimately leading to improved yields and sustainability.

Evaluating Bacillus flexus as bioremediators for ammonia removal in shrimp culture water and wastewater and characterizing microbial communities in shrimp pond sludge.

The accumulation of nitrogen compounds in shrimp farming water and effluent presents a major challenge. Ammonia is a form of nitrogen that limits shrimp growth due to its potential toxicity and effects on shrimp health and water quality. This study is aimed at identifying promising bioremediators from shrimp pond sludge to mitigate ammonia levels in both culture water and wastewater and at determining major bacterial communities in sludge using metagenomic analysis. A sludge sample was collected from a shrimp pond in Selangor, Malaysia, to isolate potential ammonia-removing bacteria. Out of 64 isolated strains, Bacillus flexus SS2 showed the highest growth in synthetic basal media (SBM) containing ammonium sulfate at a concentration of 70 mg/L as the sole nitrogen source. The strain was then incubated in SBM with varying pH levels and showed optimal growth at pH 6.5-7. After 24 h of incubation, B. flexus SS2 reduced the ammonia concentration from an initial concentration of 5 to 0.01 mg/L, indicating a 99.61% reduction rate, which was highest in SBM at pH 7. Moreover, the strain showed ammonia removal ability at concentrations ranging from 5 to 70 mg/L. Metagenomic analysis revealed that Proteobacteria was the most abundant phylum in the sludge, followed by Cyanobacteria, Actinobacteria, Chloraflexi, Firmicutes, and Campilobacterota. Bacillus flexus SS2 belongs to the Bacillota phylum and has the potential to serve as a bioremediator for removing ammonia from shrimp culture water and wastewater.

Efficacy of Single and Multi-Strain Probiotics on In Vitro Strain Compatibility, Pathogen Inhibition, Biofilm Formation Capability, and Stress Tolerance.

Compatibility of each strain in a multi-strain probiotic (MSP), along with its properties, becomes a strong base for its formulation. In this study, single-strain probiotics (SSPs) and multi-strain probiotics (MSPs) were evaluated in vitro for strain compatibility, microbial antagonism, biofilm formation capacity, and stress tolerance. Bacillus amyloliquefaciens L11, Enterococcus hirae LAB3, and Lysinibacillus fusiformis SPS11 were chosen as MSP1 candidates because they showed much stronger antagonism to Aeromonas hydrophila and Streptococcus agalactiae than a single probiotic. MSP 2 candidates were Lysinibacillus fusiformis strains SPS11, A1, and Lysinibacillus sphaericus strain NAS32 because the inhibition zone produced by MSP 2 against Vibrio harveyi and Vibrio parahaemolyticus was much higher than that produced by its constituent SSPs. MSP1 in the co-culture assay reduced (p < 0.05) A. hydrophila count from 9.89 ± 0.1 CFU mL−1 to 2.14 ± 0.2 CFU mL−1. The biofilm formation of both MSPs were significantly higher (p < 0.05) than its constituent SSPs and the pathogens. The SSPs in both MSPs generally showed resistance to high temperatures (80, 90, and 100 °C) and a wide range of pH (2 to 9). This in vitro assessment study demonstrates that MSP1 and 2 have the potential to be further explored as multi-strain probiotics on selected aquatic species.

Synthesis, Characterization and Biomedical Application of Silver Nanoparticles.

Silver nanoparticles (AgNPs) have been employed in various fields of biotechnology due to their proven properties as an antibacterial, antiviral and antifungal agent. AgNPs are generally synthesized through chemical, physical and biological approaches involving a myriad of methods. As each approach confers unique advantages and challenges, a trends analysis of literature for the AgNPs synthesis using different types of synthesis were also reviewed through a bibliometric approach. A sum of 10,278 publications were analyzed on the annual numbers of publication relating to AgNPs and biological, chemical or physical synthesis from 2010 to 2020 using Microsoft Excel applied to the Scopus publication database. Furthermore, another bibliometric clustering and mapping software were used to study the occurrences of author keywords on the biomedical applications of biosynthesized AgNPs and a total collection of 224 documents were found, sourced from articles, reviews, book chapters, conference papers and reviews. AgNPs provides an excellent, dependable, and effective solution for seven major concerns: as antibacterial, antiviral, anticancer, bone healing, bone cement, dental applications and wound healing. In recent years, AgNPs have been employed in biomedical sector due to their antibacterial, antiviral and anticancer properties. This review discussed on the types of synthesis, how AgNPs are characterized and their applications in biomedical field.

Effects of Probiotics on Growth, Survival, Water Quality and Disease Resistance of Red Hybrid Tilapia (Oreochromis spp.) Fingerlings in a Biofloc System.

Biofloc technology has shown positive effects in aquaculture, especially on the growth performance of cultured animals. The aims of this study were to evaluate the effects of adding different probiotic strains in a biofloc system on the growth performance and disease resistance of red hybrid tilapia (Oreochromis spp.). Three different probiotics (Lysinibacillus fusiformis SPS11, Bacillus amyloliquefaciens L9, and Enterococcus hirae LAB3), commercial probiotics (MG1) and a mixed probiotics (MP) combining all three strains were used in this study. The in vitro assay results showed that the mixed probiotic (MP) was able to inhibit the growth of Streptococcus agalactiae and Streptococcus iniae significantly compared to the single and commercial probiotic. The efficacy of MP was further tested in in vivo tilapia culture challenged with S. agalactiae. The best specific growth rate (3.73 ± 0.23% day-1) and feed conversion ratio (0.76 ± 0.04) were recorded in the group of biofloc with addition of MP. After being challenged with S. agalactiae, the group of biofloc with MP had significantly higher survival (83 ± 1.43%) compared to the other groups. Furthermore, the nitrogen concentration (NO2-N and NH4-N) was significantly lower in all the biofloc groups compared to the control. Hence, the addition of probiotics was able to provide beneficial effects to red hybrid tilapia culture in the biofloc system.

Influence of Symbiotic Probiont Strains on the Growth of Amphora and Chlorella and Its Potential Protections Against Vibrio spp. in Artemia.

The emerging aquaculture industry is in need of non-antibiotic-based disease control approaches to minimize the risk of antibiotic-resistant bacteria. Bacterial infections mainly caused by Vibrio spp. have caused mass mortalities of fish especially during the larval stages. The objectives of this study were to verify the potential of symbiotic probiont strains, isolated from microalgae (Amphora, Chlorella, and Spirulina) for suppressing the growth of Vibrio spp. and at the same time ascertain their abilities to enhance microalgal biomass by mutualistic interactions through microalgae-bacteria symbiosis. In addition, in vivo studies on Artemia bioencapsulated with probiont strains (single strain and mix strains) and microalgae were evaluated. The selected potential probionts were identified as Lysinibacillus fusiformis strain A-1 (LFA-1), Bacillus sp. strain A-2 (BA-2), Lysinibacillus fusiformis strain Cl-3 (LFCl-3), and Bacillus pocheonensis strain S-2 (BPS-2) using 16s rRNA. The cell densities of Amphora culture supplemented with BA-2 and Chlorella culture supplemented with LFCl-3 were higher than those of the controls. Artemia bioencapsulated with mix strains (LFA-1 + BA-2 + LFCl-3 + BPS-2) and Amphora demonstrated the highest survival rate compared to the controls, after being challenged with V. harveyi (60 ± 4%) and V. parahaemolyticus (78 ± 2%). Our study postulated that BA-2 and LFCl-3 were found to be good promoting bacteria for microalgal growth and microalgae serve as a vector to transport probiotic into Artemia. Moreover, mixture of potential probionts is beneficial for Artemia supplementation in conferring protection to Artemia nauplii against pathogenic Vibrios.

The Mechanistic Action of Biosynthesised Silver Nanoparticles and Its Application in Aquaculture and Livestock Industries.

Nanotechnology is a rapidly developing field due to the emergence of various resistant pathogens and the failure of commercial methods of treatment. AgNPs have emerged as one of the best nanotechnology metal nanoparticles due to their large surface-to-volume ratio and success and efficiency in combating various pathogens over the years, with the biological method of synthesis being the most effective and environmentally friendly method. The primary mode of action of AgNPs against pathogens are via their cytotoxicity, which is influenced by the size and shape of the nanoparticles. The cytotoxicity of the AgNPs gives rise to various theorized mechanisms of action of AgNPs against pathogens such as activation of reactive oxygen species, attachment to cellular membranes, intracellular damage and inducing the viable but non-culturable state (VBNC) of pathogens. This review will be centred on the various theorized mechanisms of actions and its application in the aquaculture, livestock and poultry industries. The application of AgNPs in aquaculture is focused around water treatment, disease control and aquatic nutrition, and in the livestock application it is focused on livestock and poultry.

The Efficacy of Moina micrura Enriched with Probiotic Bacillus pocheonensis in Enhancing Survival and Disease Resistance of Red Hybrid Tilapia (Oreochromis spp.) Larvae.

The administration of probiotics via live feeds, such as Artemia and rotifers, has gained significant attention. Moreover, indiscriminate use of antibiotics in conventional aquaculture practices in order to prevent or control disease outbreaks has resulted in the occurrence of residues and antimicrobial resistance. Thus, the application of eco-friendly feed additives, such as probiotics, as a safer alternative has received increasing attention in recent years. However, only minimal information on the administration of probiotics via freshwater cladoceran Moina micrura is available despite being commonly used for larval and post-larval feeding of freshwater crustaceans and fish. Thus, this study aimed to evaluate the application of Bacillus pocheonensis strain S2 administered via M. micrura to red hybrid tilapia (Oreochromis spp.) larvae. Bacillus pocheonensis that has been previously isolated from Spirulina sp. was subjected to preliminary in vitro evaluation of antagonistic properties. The agar well-diffusion assay revealed that this probiont could inhibit the growth of Streptococcus agalactiae and Aeromonas hydrophila. The size of inhibition zones ranged from 8.8 ± 0.2 to 18.2 ± 0.4 mm. Moina micrura was later used as a biological model in preliminary in vivo bacterial challenge assays to evaluate the efficacy of B. pocheonensis in protecting the host from diseases. Moina micrura was pre-enriched with B. pocheonensis at 104 and 106 CFU mL-1 before S. agalactiae and A. hydrophila were introduced into the culture. The study revealed that B. pocheonensis at 104 CFU mL-1 was able to significantly enhance the survival of M. micrura after being challenged with both pathogens (63 ± 3%) in comparison to the control group. The relative percentage survival (RPS) of M. micrura was highest (p < 0.05) when treated with B. pocheonensis at both concentrations 104 and 106 CFU mL-1 (38.33) after being challenged against S. agalactiae. To assess the efficacy of B. pocheonensis in protecting red hybrid tilapia against streptococcosis, the larvae were fed with either unenriched (control) Moina or probiont-enriched Moina daily for 10 days. A significantly (p < 0.05) higher survival rate (77 ± 3%) was observed in larvae fed with probiont-enriched M. micrura compared to other treatments, and the RPS was recorded at 62.90. In addition, the S. agalactiae load was suppressed in larvae fed probiont-enriched M. micrura (6.84±0.39 CFU mL-1) in comparison to the control group (7.78±0.09 CFU mL-1), indicating that the probiont might have contributed to the improvement of tilapia health and survival. This study illustrated that M. micrura was suitable to be used as a vector for probiotics in freshwater fish larvae as an alternative to hazardous antibiotics for disease control.

Antimicrobial activities of Bacillus velezensis strains isolated from stingless bee products against methicillin-resistant Staphylococcus aureus.

Infections caused by methicillin-resistant Staphylococcus aureus (MRSA) have reached epidemic proportions globally. Therefore, there is an urgent need for a continuous supply of antibiotics to combat the problem. In this study, bacteria initially identified as species belonging to the Bacillus amyloliquefaciens operational group were re-identified based on the housekeeping gene, gyrB. Cell-free supernatants (CFS) from the strains were used for antimicrobial tests using the agar well diffusion assay against MRSA and various types of pathogenic bacteria. The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and physicochemical characteristics of the CFS were determined. Based on gyrB sequence analysis, five strains (PD9, B7, PU1, BP1 and L9) were identified as Bacillus velezensis. The CFS of all B. velezensis strains showed broad inhibitory activities against Gram-negative and -positive as well as MRSA strains. Strain PD9 against MRSA ATCC 33742 was chosen for further analysis as it showed the biggest zone of inhibition (21.0 ± 0.4 mm). The MIC and MBC values obtained were 125 µl/ml. The crude antimicrobial extract showed bactericidal activity and was stable at various temperatures (40-80°C), pH (4-12), surfactants (Tween 20, Tween 80, SDS and Triton X-100) and metal ions (MgCI2, NaCI2, ZnNO3 and CuSO4) when tested. However, the crude extract was not stable when treated with proteinase K. All these properties resembled the characteristics of peptides. The antimicrobial compound from the selected strain was purified by using solvent extraction method and silica gel column chromatography. The purified compound was subjected to High Performance Liquid Chromatography which resulted in a single peak of the anti-MRSA compound being detected. The molecular weight of the anti-MRSA compound was determined by using SDS-PAGE and zymogram. The size of the purified antimicrobial peptide was approximately ~ 5 kDa. The antimicrobial peptide produced from B. velezensis strain PD9 is a promising alternative to combat the spread of MRSA infections in the future.

A Review on the Biotechnological Applications of the Operational Group Bacillus amyloliquefaciens.

Bacteria under the operational group Bacillus amyloliquefaciens (OGBa) are all Gram-positive, endospore-forming, and rod-shaped. Taxonomically, the OGBa belongs to the Bacillus subtilis species complex, family Bacillaceae, class Bacilli, and phylum Firmicutes. To date, the OGBa comprises four bacterial species: Bacillus amyloliquefaciens, Bacillus siamensis, Bacillus velezensis and Bacillus nakamurai. They are widely distributed in various niches including soil, plants, food, and water. A resurgence in genome mining has caused an increased focus on the biotechnological applications of bacterial species belonging to the OGBa. The members of OGBa are known as plant growth-promoting bacteria (PGPB) due to their abilities to fix nitrogen, solubilize phosphate, and produce siderophore and phytohormones, as well as antimicrobial compounds. Moreover, they are also reported to produce various enzymes including α-amylase, protease, lipase, cellulase, xylanase, pectinase, aminotransferase, barnase, peroxidase, and laccase. Antimicrobial compounds that able to inhibit the growth of pathogens including non-ribosomal peptides and polyketides are also produced by these bacteria. Within the OGBa, various B. velezensis strains are promising for use as probiotics for animals and fishes. Genome mining has revealed the potential applications of members of OGBa for removing organophosphorus (OPs) pesticides. Thus, this review focused on the applicability of members of OGBa as plant growth promoters, biocontrol agents, probiotics, bioremediation agents, as well as producers of commercial enzymes and antibiotics. Here, the bioformulations and commercial products available based on these bacteria are also highlighted. This review will better facilitate understandings of members of OGBa and their biotechnological applications.

Enhancement of Live Food Nutritional Status with Essential Nutrients for Improving Aquatic Animal Health: A Review.

At the present time, no artificial larval diet is capable of entirely fulfilling the dietary requirements of several larval fish and crustacean species. Zooplankton live food is the basic foundation of fish larviculture, and successful rearing of fish larvae still heavily depends on an adequate supply of nutritious live food. Despite being important, the production protocols of copepods and cladocerans (Moina) are still underdeveloped in hatcheries. Rotifers and Artemia are the most commonly used live foods. However, these live foods are evidently lacking in crucial nutrient constituents. Hence, through nutrient enrichment, live food with the nutritional profile that meets the requirements of fish larvae can be produced. With the aim to maximize the effectiveness of production to optimize profitability, it is important to evaluate and improve culture techniques for the delivery of micro- and macro-nutrients as feed supplements to larvae in aquaculture systems. Bioencapsulation and enrichment are the evolving techniques in aquaculture that are commonly employed to enhance the nutritional quality of live food by integrating nutrients into them, which subsequently improves the growth, survival, and disease resistance of the consuming hosts. This review aims to highlight some of the approaches and methods used to improve the nutritional quality of live food by modifying their nutrient composition, which could have immense promise in the enhancement of aquatic animal health.

Elucidating the Efficacy of Vaccination against Vibriosis in Lates calcarifer Using Two Recombinant Protein Vaccines Containing the Outer Membrane Protein K (r-OmpK) of Vibrio alginolyticus and the DNA Chaperone J (r-DnaJ) of Vibrio harveyi.

Recombinant cell vaccines expressing the OmpK and DnaJ of Vibrio were developed and subsequently, a vaccination efficacy trial was carried out on juvenile seabass (~5 cm; ~20 g). The fish were divided into 5 groups of 50 fish per group, kept in triplicate. Groups 1 and 2 were injected with 107 CFU/mL of the inactivated recombinant cells vaccines, the pET-32/LIC-OmpK and pET-32/LIC-DnaJ, respectively. Group 3 was similarly injected with 107 CFU/mL of inactivated E. coli BL21 (DE3), Group 4 with 107 CFU/mL of formalin killed whole cells V. harveyi, and Group 5 with PBS solution. Serum, mucus, and gut lavage were used to determine the antibody levels before all fish were challenged with V. harveyi, V. alginolyticus, and V. parahemolyticus, respectively on day 15 post-vaccination. There was significant increase in the serum and gut lavage antibody titers in the juvenile seabass vaccinated with r-OmpK vaccine. In addition, there was an up-regulation for TLR2, MyD88, and MHCI genes in the kidney and intestinal tissues of r-OmpK vaccinated fish. At the same time, r-OmpK triggered higher expression level of interleukin IL-10, IL-8, IL-1ß in the spleen, intestine, and kidney compared to r-DnaJ. Overall, r-OmpK and r-DnaJ triggered protection by curbing inflammation and strengthening the adaptive immune response. Vaccinated fish also demonstrated strong cross protection against heterologous of Vibrio isolates, the V. harveyi, V. alginolyticus, and V. parahaemolyticus. The fish vaccinated with r-OmpK protein were completely protected with a relative per cent of survival (RPS) of 90 percent against V. harveyi and 100 percent against V. alginolyticus and V. parahaemolyticus. A semi-quantitative PCR detection of Vibrio spp. from the seawater containing the seabass also revealed that vaccination resulted in reduction of pathogen shedding. In conclusion, our results suggest r-OmpK as a candidate vaccine molecule against multiple Vibrio strain to prevent vibriosis in marine fish.

Characterization of Enterococcus hirae Isolated from the Intestine of Seabass (Lates Calcarifer) as a New Potential Probiotic against Pathogenic Vibrios.

In this study, we aimed to isolate, identify and characterize lactic acid bacteria (LAB) from the intestine of juvenile seabass (Lates calcarifer) as a new potential probiotic. Four strains of LABs were isolated from the intestines of ten healthy seabass juveniles. In the in vitro screening process using spot lawn assay, one isolate labeled as LAB3 showed inhibitory activity against Vibrio harveyi (ATCC 35,084). Strain LAB3 was determined to belong to the gram positive bacteria group with cocci shape and was identified as Enterococcus hirae using 16S rDNA analysis. This bacterium was able to grow at pH ranging from pH 2 to 10 with the best growth at pH 7. This strain was also able to grow at 0-4% NaCl after 24 h incubation and grew best at 1.5% NaCl. Enterococcus hirae strain LAB3 of the present study is worthy to be further characterized as a potential probiotic for use in seabass culture.

Quorum sensing is required for full virulence of Vibrio campbellii towards tiger grouper (Epinephelus fuscoguttatus) larvae.

The link between quorum sensing in Vibrio campbellii and its virulence towards tiger grouper (Epinephelus fuscoguttatus) was investigated using V. campbellii wild type and quorum-sensing mutants with inactive quorum sensing or constitutively maximal quorum-sensing activity, and signal molecule synthase mutants. The results showed that wild-type V. campbellii is pathogenic to grouper larvae, causing more than 50% mortality after 4 days of challenge. Furthermore, the mortality of larvae challenged with the mutant with maximally active quorum sensing was significantly higher than that of larvae challenged with the wild type, whereas a higher survival was observed in the larvae challenged to the mutant with a completely inactive quorum-sensing system. Grouper larvae challenged with either the signal molecule synthase triple mutant, the harveyi autoinducer-1 (HAI-1) synthase mutant and the autoinducer-2 (AI-2) synthase mutant showed higher survival than larvae challenged with the wild type. In contrast, larvae challenged with the cholerae autoinducer-1 (CAI-1) synthase mutant showed high mortality. This indicates that HAI-1 and AI-2, but not CAI-1, are required for full virulence of V. campbellii towards grouper larvae. Our data suggest that quorum-sensing inhibition could be an effective strategy to control V. campbellii infections in tiger grouper.

Contributions of tropodithietic acid and biofilm formation to the probiotic activity of Phaeobacter inhibens.

BACKGROUND: The probiotic bacterium Phaeobacter inhibens strain S4Sm, isolated from the inner shell surface of a healthy oyster, secretes the antibiotic tropodithietic acid (TDA), is an excellent biofilm former, and increases oyster larvae survival when challenged with bacterial pathogens. In this study, we investigated the specific roles of TDA secretion and biofilm formation in the probiotic activity of S4Sm. RESULTS: Mutations in clpX (ATP-dependent ATPase) and exoP (an exopolysaccharide biosynthesis gene) were created by insertional mutagenesis using homologous recombination. Mutation of clpX resulted in the loss of TDA production, no decline in biofilm formation, and loss of the ability to inhibit the growth of Vibrio tubiashii and Vibrio anguillarum in co-colonization experiments. Mutation of exoP resulted in a ~60% decline in biofilm formation, no decline in TDA production, and delayed inhibitory activity towards Vibrio pathogens in co-colonization experiments. Both clpX and exoP mutants exhibited reduced ability to protect oyster larvae from death when challenged by Vibrio tubiashii. Complementation of the clpX and exoP mutations restored the wild type phenotype. We also found that pre-colonization of surfaces by S4Sm was critical for this bacterium to inhibit pathogen colonization and growth. CONCLUSIONS: Our observations demonstrate that probiotic activity by P. inhibens S4Sm involves contributions from both biofilm formation and the production of the antibiotic TDA. Further, probiotic activity also requires colonization of surfaces by S4Sm prior to the introduction of the pathogen.

Effects of some dietary crude plant extracts on the growth and gonadal maturity of Nile tilapia (Oreochromis niloticus) and their resistance to Streptococcus agalactiae infection.

A 90-day feeding trial was conducted on the growth performance, feeding efficacy, body indices, various hematological and plasma biochemical parameters, and histopathological examination of the gonads from male and female Nile tilapia fingerlings when fed different crude plant extracts from Cinnamomum camphora, Euphorbia hirta, Azadirachta indica, or Carica papaya at 2 g kg(-1) compared to a control diet. This was followed by a 14-day challenge to Streptococcus agalactiae. All treatments were triplicated, and each treatment consisted of 30 fish. Results showed that C. papaya extracts were the most effective at delaying gonadal maturation to both male and female tilapia, as well as significantly increasing (P < 0.05) growth performance compared to the control treatment. Similarly, dietary C. camphora and E. hirta extracts also significantly improved growth, while no significant growth effect was detected between the A. indica and control treatments (P > 0.05). Further, crude body lipid was lower in the C. camphora, E. hirta and C. papaya treatments, but was only significantly lower for the E. hirta treatment compared to the control. Meanwhile, none of the hematological or biochemical parameters were significantly affected, although plasma ALT was significantly lower for tilapia fed A. indica compared to the control. After the 14-day bacterial challenge, tilapia fed C. camphora supplementation had significantly higher survival, compared to the control, but was not significantly higher than the other supplemented diets. Results indicate that dietary C. papaya extract can significantly promote growth and delay gonadal maturation to both male and female tilapia, while C. camphora was the most effective prophylactic to S. agalactiae and may be a cost-effective and eco-friendly alternative to antibiotics.

Screening and evaluation of local bacteria isolated from shellfish as potential probiotics against pathogenic Vibrios.

The present study was carried out to isolate, screen and evaluate potential candidates of local bacteria isolated from tiger shrimp Penaeus monodon and slipper cupped oysters Crassostrea iredalei as probiotics in shellfish aquaculture. A total of 144 of bacteria were successfully isolated from the intestine and stomach of 20 tails of healthy adult tiger shrimp P. monodon, while 136 were successfully isolated from the digestive tract, gills and inner shells of 10 healthy adult C. iredalei. The number of potential isolates was narrowed down to two from tiger shrimp, and one from slipper cupped oyster after in vitro screening assays. The three isolates, labeled as G11, I24 and S66, were identified as Virgibacillus sp., Bacillus sp. and Exiquobacterium sp., respectively, using 16S rDNA gene analysis. The antagonistic ability of the isolates towards Vibrio alginolyticus and Vibrio harveyi were conducted in stagnant and liquid modes via spot lawn and broth co-culture assay, respectively. In these assays, all the potential probionts were inhibitory to both pathogenic vibrios. In the in-vivo assay, Artemia was used as host and treated with different concentrations of potential probionts (10(4), 10(6) and 10(8) CFU ml(-1)), and challenged with V. alginolyticus and V. harveyi at 105 CFU ml(-1), respectively. Artemia treated with probiont G11 at all concentrations and challenged with V. alginolyticus had increased survival (70 ? 80 %), which was significantly higher as compared with group with only the pathogen (20 %). Meanwhile, probiont I24 increased the survival of Artemia by 70 % at a concentration of 10(8) CFU ml(-1) after being challenged with V. alginolyticus and Artemia treated with 10(6) CFU ml(-1) of probiont S66 had increased survival of 90% after being challenged with V. harveyi. Thus, the three isolates might have potential applications as probiotics in shellfish aquaculture against vibriosis. ?

The effects of dietary inclusion of garlic on growth performance and disease resistance of African catfish (Clarias gariepinus) fingerlings against Aeromonas hydrophila infection.

The present study aimed to investigate the effects of dietary inclusion of garlic (Allium sativum) peels and cloves on the growth performance and disease resistance of African catfish (Clarias gariepinus) fingerlings against Aeromonas hydrophila infection. Seven isonitrogenous (36% protein) experimental diets were formulated to contain graded levels of garlic (peels and cloves) at 0, 10, 20 and 30g.kg-1. Fish were fed twice a day for 12 weeks. The results demonstrated that no significant differences were observed with respect to growth performance or feed utilization efficiency (i.e., body weight gain (WG) (133 ? 0.3g), specific growth rate (SGR) (2.23 ? 0.04%), and feed conversion ratio (FCR) (1.00 ? 0.2g) of fish fed with different inclusion levels of garlic peels and cloves as compared to control group. The plasma biochemical results showed higher total protein, albumin and globulin content in control group (T1) as compared to the experimental groups, but these results were not significant. After the fish were challenged with A. hydrophila, low survival (13 %) was found in control group which was significantly lower as compared to all the treatment groups (> 35 %). Meanwhile, the highest survival (64%) was observed for fish fed with garlic cloves at 20g.kg-1 (T3). The results showed that inclusion of garlic cloves at 20 g.kg(-1) could enhance the resistance of African catfish fingerlings to A. hydrophila infection. ??

The effects of dietary inclusion of garlic on growth performance and disease resistance of African catfish (Clarias gariepinus) fingerlings against Aeromonas hydrophila infection.

The present study aimed to investigate the effects of dietary inclusion of garlic (Allium sativum) peels and cloves on the growth performance and disease resistance of African catfish (Clarias gariepinus) fingerlings against Aeromonas hydrophila infection. Seven isonitrogenous (36% protein) experimental diets were formulated to contain graded levels of garlic (peels and cloves) at 0, 10, 20 and 30g.kg-1. Fish were fed twice a day for 12 weeks. The results demonstrated that no significant differences were observed with respect to growth performance or feed utilization efficiency (i.e., body weight gain (WG) (133 ? 0.3g), specific growth rate (SGR) (2.23 ? 0.04%), and feed conversion ratio (FCR) (1.00 ? 0.2g) of fish fed with different inclusion levels of garlic peels and cloves as compared to control group. The plasma biochemical results showed higher total protein, albumin and globulin content in control group (T1) as compared to the experimental groups, but these results were not significant. After the fish were challenged with A. hydrophila, low survival (13 %) was found in control group which was significantly lower as compared to all the treatment groups (> 35 %). Meanwhile, the highest survival (64%) was observed for fish fed with garlic cloves at 20g.kg(-1) (T3). The results showed that inclusion of garlic cloves at 20 g.kg(-1) could enhance the resistance of African catfish fingerlings to A. hydrophila infection. ??