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Fish & Shellfish Immunology May 2023In recent years, the widespread use of antibiotics in intensive grouper mariculture has resulted in the ineffectiveness of antibiotic treatment, leading to an increasing...
Screening of host gut-derived probiotics and effects of feeding probiotics on growth, immunity, and antioxidant enzyme activity of hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂).
In recent years, the widespread use of antibiotics in intensive grouper mariculture has resulted in the ineffectiveness of antibiotic treatment, leading to an increasing incidence of diseases caused by bacteria, viruses, and parasites, causing serious economic losses. Hence, it is crucial to develop alternative strategies to antibiotics for healthy and sustainable development of the mariculture industry. Here, we aimed to screen host gut-derived probiotics and evaluate its effects on growth and immunity of grouper. In this study, 43 bacterial strains were isolated from the intestine of the hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂), and a potential probiotic strain G1-26, which can efficiently secrete amylase, protease, and lipase, was obtained using different screening mediums. Based on 16S rDNA sequencing, the potential probiotic strain G1-26 was identified as Vibrio fluvialis. The results of a biological characteristic evaluation showed that V. fluvialis G1-26 could grow at 25-45 °C, pH 5.5-7.5, salinity 10-40, and bile salt concentration 0-0.030%, and produce amylase, lipase, and protease under different culture conditions. Additionally, V. fluvialis G1-26 is sensitive to many antibiotics and does not exhibit aquatic biotoxicity. Subsequently, hybrid groupers were fed diets containing V. fluvialis G1-26 at different concentrations (0, 10, 10, and 10 CFU/g) for 60 d. The results showed that V. fluvialis G1-26 at 10 CFU/g did not significantly affect the growth performance of the hybrid grouper (P > 0.05). V. fluvialis G1-26 supplementation at 10 and 10 CFU/g significantly promoted the relative expression of immune-related genes in hybrid groupers (TLR3, TLR5, IL-1β, IL-8, IL-10, CTL, LysC, TNF-2, and MHC-2) and improved the activities of alkaline phosphatase, acid phosphatase, total superoxide dismutase, and total protein in the liver. In conclusion, V. fluvialis G1-26, a potential probiotic strain isolated from the intestine of the hybrid grouper, can be used as an effective immunopotentiator at an optimal dose of 10 CFU/g diet. Our results provide a scientific basis for the development and utilization of probiotics in the grouper mariculture industry.
Topics: Animals; Antioxidants; Bass; Diet; Probiotics; Peptide Hydrolases; Amylases; Lipase; Animal Feed
PubMed: 36966895
DOI: 10.1016/j.fsi.2023.108700 -
Frontiers in Microbiology 2019Food safety has become an important public health issue worldwide. However, conventional methods for detection of food-borne pathogens are complicated, and...
Food safety has become an important public health issue worldwide. However, conventional methods for detection of food-borne pathogens are complicated, and labor-intensive. Moreover, the sensitivity is often low, and it is difficult to achieve high-throughput detection. This study developed a TaqMan real-time polymerase chain reaction (PCR) assay for the simultaneous detection and quantification of 12 common pathogens in a single reaction, including , β hemolyticus, spp., , and in food and drinking water. Based on published sequence data, specific primers, and fluorescently-labeled hybridization probes were designed targeting based on the virulence genes of the 12 pathogens, and these primers and probes were optimized to achieve consistent reaction conditions. The assay was evaluated using 106 pure bacterial culture strains. There was no cross-reaction among the different pathogens. The analytical sensitivity was 1 copy/μL for , β spp., , and , 10 copies/μL for , and , respectively. The limit of detection (LOD) was 296, 500, 177, 56, 960, 830, 625, 520, 573, 161, 875, and 495 CFU/mL for , β spp., , and , respectively. The limit of detection for the assay in meat samples was 10 CFU/g for and 10 CFU/g for other 11 strains. Together, these results indicate that the optimized TaqMan real-time PCR assay will be useful for routine detection of pathogenic bacteria due to its rapid analysis, low cost, high-throughput, high specificity, and sensitivity.
PubMed: 30814987
DOI: 10.3389/fmicb.2019.00222 -
3 Biotech Jun 2016The diversity of some of the culturable microorganisms associated with marine flora and fauna collected off Vizhinjam and Mulloor coast of South India was evaluated and...
The diversity of some of the culturable microorganisms associated with marine flora and fauna collected off Vizhinjam and Mulloor coast of South India was evaluated and their bioactive production potential determined. From a total of 24 bacteria, 4 actinomycetes and 8 fungi isolated from diverse marine sources, five bacterial species-BLM3, BSP2, BCS1, BCS4 and BMA6 showed inhibitory activity against at least one of the tested pathogens viz., Klebsiella pneumonia KU1, Pseudomonas aeruginosa VL3, Salmonella enterica typhimurium MTCC 98, Escherichia coli MTCC 40, Micrococcus luteus MTCC 105, Staphylococcus simulans MTCC 3610, Proteus vulgaris MTCC 426, Vibrio fluvialis, Vibrio sp. P3a and Vibrio sp. P3b. The isolated actinomycetes and fungi did not produce significant inhibition zones against the tested pathogens; however, the macroalgal isolated actinomycetes strain AMA1 produced reddish pigment in Starch Casein medium which remained stable till the stationary phase of growth. The marine sediment isolate BCS4, identified as Bacillus sp. showed wide spectrum of activity against the tested Gram positive bacteria, S. simulans MTCC 3610 and Gram negative bacteria, Proteus vulgaris with zone of inhibitions of 25 and 11 mm respectively. Better extraction of the bioactive compound was obtained with ethyl acetate when compared with methanol, benzene and hexane and TLC analysis revealed the presence of an active compound. The 16SrRNA sequencing confirmed the potent strain belong to Bacillus sp. and hence designated Bacillus sp. BCS4.
PubMed: 28330077
DOI: 10.1007/s13205-015-0318-1 -
Complete genomic sequence of strain IDH5335 isolated from a patient with diarrhea in Kolkata, India.Microbiology Resource Announcements Dec 2023We isolated a strain (IDH5335) from a stool sample collected from a patient with diarrhea. In this announcement, we report the complete genomic sequence of this...
We isolated a strain (IDH5335) from a stool sample collected from a patient with diarrhea. In this announcement, we report the complete genomic sequence of this organism, which was obtained by combining Illumina and Oxford Nanopore sequencing data.
PubMed: 37943041
DOI: 10.1128/MRA.00707-23 -
Microbial Drug Resistance (Larchmont,... Apr 2016The aim of this study was to evaluate the presence of Vibrio isolates recovered from four different fish pond facilities in Benin City, Nigeria, determine their...
The aim of this study was to evaluate the presence of Vibrio isolates recovered from four different fish pond facilities in Benin City, Nigeria, determine their antibiogram profiles, and evaluate the public health implications of these findings. Fish pond water samples were collected from four sampling sites between March and September 2014. A total of 56 samples were collected and screened for the isolation of Vibrio species using standard culture-based methods. Polymerase chain reaction (PCR) was used to confirm the identities of the Vibrio species using the genus-specific and species-specific primers. Vibrio species were detected at all the study sites at a concentration on the order of 10(3) and 10(6) CFU/100 ml. A total of 550 presumptive Vibrio isolates were subjected to PCR confirmation. Of these isolates, 334 isolates tested positive, giving an overall Vibrio prevalence rate of 60.7%. The speciation of the 334 Vibrio isolates from fish ponds yielded 32.63% Vibrio fluvialis, 20.65% Vibrio parahaemolyticus, 18.26% Vibrio vulnificus, and 28.44% other Vibrio species. In all, 167 confirmed Vibrio isolates were selected from a pool of 334 confirmed Vibrio isolates for antibiogram profiling. The susceptibility profiles of 20 antimicrobial agents on the isolates revealed a high level of resistance for AMP(R), ERY(R), NAL(R), SUL(R), TMP(R), SXT(R), TET(R), OTC(R), and CHL(R). The percentage of multiple drug resistance Vibrio isolates was 67.6%. The multiple antibiotic resistance index mean value of 0.365 for the Vibrio isolates found in this study indicated that the Vibrio isolates were exposed to high-risk sources of contamination when antibiotics were frequently used. The resistant Vibrio strains could be transmitted through the food chain to humans and therefore constitutes a risk to public health.
Topics: Anti-Bacterial Agents; Aquaculture; Drug Resistance, Multiple, Bacterial; Environment; Environmental Microbiology; Nigeria; Ponds; Public Health; Vibrio
PubMed: 26540391
DOI: 10.1089/mdr.2015.0169 -
Frontiers in Microbiology 2023High hydrostatic pressure (HHP) regulated gene expression is one of the most commonly adopted strategies for microbial adaptation to the deep-sea environments....
High hydrostatic pressure (HHP) regulated gene expression is one of the most commonly adopted strategies for microbial adaptation to the deep-sea environments. Previously we showed that the HHP-inducible trimethylamine N-oxide (TMAO) reductase improves the pressure tolerance of deep-sea strain QY27. Here, we investigated the molecular mechanism of HHP-responsive regulation of TMAO reductase TorA. By constructing and deletion mutants, we demonstrated that the two-component regulator TorR and sensor TorS are responsible for the HHP-responsive regulation of . Unlike known HHP-responsive regulatory system, the abundance of and was not affected by HHP. Complementation of the Δ mutant with TorS altered at conserved phosphorylation sites revealed that the three sites were indispensable for substrate-induced regulation, but only the histidine located in the alternative transmitter domain was involved in pressure-responsive regulation. Taken together, we demonstrated that the induction of TMAO reductase by HHP is mediated through the TorRS system and proposed a bifurcation of signal transduction in pressure-responsive regulation from the substrate-induction. This work provides novel knowledge of the pressure regulated gene expression and will promote the understanding of the microbial adaptation to the deep-sea HHP environment.
PubMed: 38029070
DOI: 10.3389/fmicb.2023.1291578 -
Frontiers in Microbiology 2017High hydrostatic pressure (HHP) exerts severe effects on cellular processes including impaired cell division, abolished motility and affected enzymatic activities....
High hydrostatic pressure (HHP) exerts severe effects on cellular processes including impaired cell division, abolished motility and affected enzymatic activities. Transcriptomic and proteomic analyses showed that bacteria switch the expression of genes involved in multiple energy metabolism pathways to cope with HHP. We sought evidence of a changing bacterial metabolism by supplying appropriate substrates that might have beneficial effects on the bacterial lifestyle at elevated pressure. We isolated a piezosensitive marine bacterium strain QY27 from the South China Sea. When trimethylamine -oxide (TMAO) was used as an electron acceptor for energy metabolism, QY27 exhibited a piezophilic-like phenotype with an optimal growth at 30 MPa. Raman spectrometry and biochemistry analyses revealed that both the efficiency of the TMAO metabolism and the activity of the TMAO reductase increased under high pressure conditions. Among the two genes coding for TMAO reductase catalytic subunits, the expression level and enzymatic activity of TorA was up-regulated by elevated pressure. Furthermore, a genetic interference assay with the CRISPR-dCas9 system demonstrated that TorA is essential for underpinning the improved pressure tolerance of QY27. We extended the study to type strain ATCC33809 and observed the same phenotype of TMAO-metabolism improved the pressure tolerance. These results provide compelling evidence for the determinant role of metabolism in the adaption of bacteria to the deep-sea ecosystems with HHP.
PubMed: 29375513
DOI: 10.3389/fmicb.2017.02646 -
Foods (Basel, Switzerland) Mar 2023Previous research has shown that freshwater edible fish imported into Australia are not compliant with Australian importation guidelines and as a result may be high risk...
Previous research has shown that freshwater edible fish imported into Australia are not compliant with Australian importation guidelines and as a result may be high risk for bacterial contamination. In the present study, the outer surface of imported freshwater fish were swabbed, cultured, confirmatory tests performed and antimicrobial patterns investigated. Channidae fish (Sp. A/n = 66) were contaminated with zoonotic sp./ (n = 1/66) and other bacteria implicated in cases of opportunistic human infection, these being sp. (including . and (n = 34/66)); sp. (n = 32/66); (n = 27/66) and (n = 3/66). Pangasiidae fish (Species B/n = 47) were contaminated with zoonotic (n = 10/47); sp. (n = 6/47) and environmental bacteria sp. (n = 3/47). One sample was resistant to all antimicrobials tested and is considered to be Methicillin Resistant . Mud, natural diet, or vegetation identified in Sp. A fish/or packaging were significantly associated with the presence of spp. The study also showed that visibly clean fish (Sp. B) may harbour zoonotic bacteria and that certain types of bacteria are common to fish groups, preparations, and contaminants. Further investigations are required to support the development of appropriate food safety recommendations in Australia.
PubMed: 36981215
DOI: 10.3390/foods12061288 -
International Journal of Food... Jan 2018The aim of this research was to evaluate the feasibility of PCR-DGGE and Reverse Transcriptase-PCR-DGGE techniques for rapid detection of Vibrio species in foods....
The aim of this research was to evaluate the feasibility of PCR-DGGE and Reverse Transcriptase-PCR-DGGE techniques for rapid detection of Vibrio species in foods. Primers GC567F and 680R were initially evaluated for amplifying DNA and cDNA of ten references Vibrio species by PCR method. The GC-clamp PCR amplicons were separated according to their sequences by the DGGE using 10% (w/v) polyacrylamide gel containing 45-70% urea and formamide denaturants. Two pair of Vibrio species, which could not be differentiated on the gel, was Vibrio fluvialis - Vibrio furnissii and Vibrio parahaemolyticus - Vibrio harveyi. To determine the detection limit, in the community of 10 reference strains containing the same viable population, distinct DNA bands of 3 species; Vibrio cholerae, Vibrio mimicus and Vibrio alginolyticus were consistently observed by PCR-DGGE technique. In fact, 5 species; Vibrio cholerae, Vibrio mimicus, Vibrio alginolyticus, Vibrio parahaemolyticus and Vibrio fluvialis consistently observed by Reverse Transcriptase-PCR-DGGE. In the community containing different viable population increasing from 10 to 10CFU/mL, PCR-DGGE analysis only detected the two most prevalent species, while RT-PCR-DGGE detected the five most prevalent species. Therefore, Reverse Transcriptase-PCR-DGGE was also selected for detection of various Vibrio cell conditions, including viable cell (VC), injured cells from frozen cultures (IVC) and injured cells from frozen cultures with pre-enrichment (PIVC). It was found that cDNA band of all cell conditions gave the same migratory patterns, except that multiple cDNA bands of Plesiomonas shigelloides under IVC and PIVC conditions were found. When Reverse Transcriptase-PCR-DGGE was used for detecting Vibrio parahaemolyticus in the pathogen-spiked food samples, Vibrio parahaemolyticus could be detected in the spiked samples containing at least 10CFU/g of this pathogen. The results obtained also corresponded to standard method (USFDA, 2004). In comparison with the detection of the Vibrio profiles in fourteen food samples using standard method, Reverse Transcriptase-PCR-DGGE resulted in 100%, 75% and 50% similarity in 3, 1 and 6 food samples, respectively.
Topics: DNA Primers; Denaturing Gradient Gel Electrophoresis; Food Microbiology; Nucleic Acid Amplification Techniques; Polymerase Chain Reaction; RNA-Directed DNA Polymerase; Reverse Transcriptase Polymerase Chain Reaction; Vibrio alginolyticus; Vibrio cholerae; Vibrio mimicus; Vibrio parahaemolyticus
PubMed: 29111407
DOI: 10.1016/j.ijfoodmicro.2017.10.014 -
Journal of Food Science and Technology Apr 2017This study examined the antibacterial activity of on multiple antibiotic resistant (MAR) and isolated from shrimps. The ethanol extract of antibacterial properties...
This study examined the antibacterial activity of on multiple antibiotic resistant (MAR) and isolated from shrimps. The ethanol extract of antibacterial properties was assessed using the agar diffusion method. Survival of test organisms in shrimp meat using different concentrations of was done using standard method. The quantitative and qualitative phytochemical tests of extract were determined. The ethanol extract had antimicrobial activities on the test organisms showing 20.00 ± 0.0 and 23.00 ± 0.0 mm zone of inhibition on and respectively. completely decreased microbial load of and in 150 and 60 min, respectively. The phytochemical screening for the ethanol extract reported phenol, alkaloids, tannin, saponin, anthraquinone flavonoid and cardiac glycoside as 51.76, 26.60, 6.76, 54.33, 30.35 89.65 and 18.23 mg/100 g, respectively. This study reveals the antibacterial property of on the MAR species.
PubMed: 28416870
DOI: 10.1007/s13197-017-2543-6