Selection of lactic acid bacteria as candidate probiotics and in vivo test on Artemia nauplii (original) (raw)
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Diseases of Aquatic Organisms, 2015
Vibrio alginolyticus is an opportunistic pathogen which may affect different aquatic organisms. The aim of this study was to assess the probiotic properties and the protective mode of action of Lactobacillus pentosus H16 against V. alginolyticus 03/8525, through in vitro and in vivo studies using Artemia franciscana (hereafter Artemia). This strain showed antimicrobial activity against V. alginolyticus 03/8525 and Aeromonas salmonicida subsp. salmonicida ATCC33658 possibly related to lactobacilli organic acid production. It was able to survive at high rainbow trout bile concentrations and showed high selective adhesion to rainbow trout mucus (1.2 × 10 5 ± 8.0 × 10 3 cells cm −2). H16 outcompeted V. alginolyticus 03/8525 and A. salmonicida subsp. salmonicida ATCC33658, greatly reducing their adherence to rainbow trout mucus (64.8 and 74.1%, respectively). Moreover, H16 produced a cell-bound biosurfactant which caused an important decrease in the surface tension. H16 also protected Artemia nauplii against mortality when it was administered previous to V. alginolyticus 03/8525 inoculation. Furthermore, H16 bioencapsulated in Artemia, suggesting that it is possible to use live carriers in its administration. We conclude that the ability of L. pentosus H16 to selectively adhere to mucosal surfaces and produce cell-bound biosurfactants, displacing pathogenic strains, in addition to its antimicrobial activity, confer H16 competitive advantages against pathogens as demonstrated in in vivo challenge experiments. Thus, L. pentosus H16, a marine bacterium from the intestinal tract of hake, is an interesting probiotic for Artemia culture and also has the potential to prevent vibriosis in other aquaculture activities such as larvae culture and fish farming.
Cell free extracts of four strains of Lactic acid bacteria (LAB) viz. Lactobacillus. acidophilus, Streptococcus.cremoris, Lactobacillus bulgaricus-56 and Lactobacillus bulgaricus-57 inhibited growth of Vibrio alginolyticus in nutrient broth. The antagonism of LAB to Vibrio alginolyticus was further confirmed by streak plating wherein suppression of growth of Vibrio was obtained. Juveniles of Penaeus indicus (average weight 0.985 ± 0.1 g) on administering orally a moist feed base containing 5 × 10 6 cells•g of the four LAB probionts for a period of four weeks showed better survival (56 to 72%) when challenged with V. alginolyticus by intra-muscular injection of 0.1 ml containing 3 × 10 9 cells•ml. Animals maintained on a diet devoid of bacterial biomass exhibited 80% mortality. No external or internal pathological changes were observed in shrimp fed with the LAB incorporated diets. Results showed inhibition of V. alginolyticus by LAB and stimulation of the non-specific immune response resulting in resistance to disease in the shrimp fed on LAB incorporated diets.
IOP Conference Series: Earth and Environmental Science, 2020
Acute hepatopancreatic necrosis disease has been a major challenge faced by white shrimp farmers in many shrimp-producing countries inducing Indonesia. The etiological agent of this disease was a virulent strain of Vibrio parahaemolyticus. However, a technology to treat or to prevent the disease infection has not been established yet, and probiont could be a potential approach for this disease. Thus, this study aimed at screening 35 lactic acid bacteria (LAB) that originated from the gastrointestinal tract of wild white shrimp for the production of antivibrio compounds using an agar well diffusion. The result showed that three LAB strains (LAB-1, LAB-2, and LAB-3) showed antivibrio activity indicated by the formation of clearance zone (diameter of >10mm) on the diffusion agar plate. When the extracellular products (ECP) were incubated with proteinase K, the antivibrio activity was disappeared which indicated that the antimicrobial compounds were bacteriocin-like inhibitory substa...
Frontiers in Marine Science, 2014
The probiotic Lactobacillus rhamnosus IMC 501® (Synbiotec Srl, Camerino, Italy) has demonstrated a positive effect in growth and early development of some fish species as clownfish (Amphiprion ocellaris, Cuvier 1830) and zebrafish (Danio rerio, Hamilton-Buchanan, 1822). The probiotic effects in A. ocellaris were an increase in body weight, a metamorphosis and skeletal head development acceleration and a reduction in the response to general stress . Besides this, an acceleration of the backbone calcification and gonadal differentiation were described in D. rerio (Avella et al. 2012). Despite those investigations, there is a lack of knowledge on the survival of L. rhamnosus in seawater and the bioencapsulation efficiency in Artemia. In the present study, the survival of L. rhamnosus in seawater and the efficiency of the probiotic bioencapsulation in 24h Artemia metanauplii were assessed. In a first experiment, 1 gr/l of probiotic bacteria was resuspended in 50 ml seawater and incubated in the conditions used for Artemia culture (26°C and gentle aeration). Samples were taken at 0, 15, 30, 60 min and at 3, 6, 24 and 30h. Serial dilutions were plated on selective MRS-Agar for probiotic bacteria isolation and on MRSA-B Agar (MRS-Agar supplemented by aniline blue for total CFU staining) . Plates were incubated for 7 days at 30°C in obscurity and CFU counted. In the bioencapsulation experiment, two conditions were tested: AS, Artemia standard (negative control, and AP, Artemia supplemented by probiotic (positive control, ). For AP bioencapsulation 30,000 Artemia 24h-metanauplii were suspended in 300 ml SSW and 1gr/l of probiotic was added. Samples were taken at 0 (after 15 min of bioencapsulation), 0.5, 1, 3, 6 and 24h. For that, the samples were filtered in a 125 µm mesh, washed and homogenized for microbiological analysis. Serial dilutions were plated on Marine Agar, TCBS Agar and MRSA-B Agar for quantification of total bacteria, total Vibrionaceae and lactic-acid bacteria, respectively. All experiments were performed in duplicate. The results obtained in the first test showed that the L. rhamnosus was able to survive in seawater during the whole experiment (30h), maintaining densities of 10e7 CFU/ml during the first 6h although decreasing
2021
Supplementation with mixed probiotic in aquaculture has been proven to benefit the hosts as disease resistance tool. In this study, a mixed probiotic which consisted of three isolated strains (Lysinibacillus fusiformis strain SPS11, A2, and Bacillus megaterium strain I24) was formulated for the in vitro assays against Vibrio alginolyticus and in vivo preliminary study towards Artemia nauplii. These strains showed antagonism activities against V. alginolyticus in in vitro assay. An increase in biofilm formation of this mixed probiotic was observed which indicated that the strains could work synergistically with each other to confer benefits to the hosts. Enrichment of Artemia nauplii with the formulated mixed probiotic was done to investigate its role in enhancing resistance against the V. alginolyticus. Artemia nauplii were cultured in two different concentrations of mixed probiotic (106 and 108 CFU mL-1) and challenged via immersion method. The mixed probiotic at both concentration...
Pure and Applied Biology, 2012
In this study, the Vibrio probioticus was isolated from the gut of female Portunus pleagicus, identified through 16S rDNA gene sequencing, and was evaluated for its probiotic features. The probiont was tested in coculture assay at various concentrations against three target shellfish pathogens including V. harveyi, V. parahaemolyticus and Pseudoalteromonas piscicida. In a co-culture assay it was apparent that the V. probioticus had shown to outcompete the growth of tested pathogenic bacteria due to its antibacteriocin properties. Different ratios produced different effect on tested pathogens, however, the highest ratio (pathogen:probiont) was more effective in suppressing the pathogen. A maximum inhibition growth was recorded for V. harveyi and P. piscicida. The probionts could further be tested in vivo culture environment to understand the effectiveness in microbial control of P. pelagicus larviculture and other aquaculture species.
International journal of pharma and bio sciences, 2014
The present work was conducted to isolate the gut bacteria of Artemia and to use them as probiotics to control the bacterial pathogens in aquaculture systems. In the present study, two of bacterial species were isolated from the gut of Artemia which was collected from Puthalam saltpan (Kanyakumari, India). Among the gut isolates of Artemia, best two species, Alteromonas sp. and Acetobacterium sp. were selected. These two isolates were tested for its efficiency and antagonistic activity against Vibrio harveyi and Alteromonas sp. Artemia gut isolates, when introduced in to the gut of adult Artemia along with pathogenic bacteria, they multiplied and colonized faster and seen in higher number than the pathogenic bacteria . During the present study it was found that Acetobacterium sp. effectively controlled the growth of V. harveyi and Alteromonas sp. than Aeromonas sp. Hence this study adds more towards the use of gut isolates of Artemia as probiotics and on controlling the bacterial pa...
World Journal of Microbiology and Biotechnology, 2012
Two potential probiotics Bacillus subtilis and Lactobacillus plantarum were evaluated for use in aquaculture as preventive measures against vibriosis. In vitro evaluation of the probiotics using co-culture assays with the pathogen Vibrio anguillarum and testing for the production of antibacterial substances showed the presence of antagonism and confirmed the production of antibacterial substances. Both potential probiotics were administered to the live fish feed Artemia franciscana nauplii, offering protection against a subsequent challenge of the nauplii with the fish pathogen V. anguillarum, with best survival rates of the nauplii and the most efficient protection offered by B. subtilis. Nauplii enriched with B. subtilis were further used to evaluate the protection of sea bass larvae against vibriosis. The untreated group of fish challenged with V. anguillarum presented low survival of 36.7 %, while the fish treated with nauplii enriched with the probiotic B. subtilis showed significantly increased survival rates of 86.7 % after challenge with the pathogen. The survival of healthy unchallenged fish treated with the probiotic was not significantly different from control unchallenged fish (90-94 %). Our results indicate that B. subtilis is a probiotic suitable to be used for the prevention of vibriosis in fish larvae and can be safely administered through their live feed Artemia nauplii.
BMC Microbiology, 2013
Background The microorganisms intended for use as probiotics in aquaculture should exert antimicrobial activity and be regarded as safe not only for the aquatic hosts but also for their surrounding environments and humans. The objective of this work was to investigate the antimicrobial/bacteriocin activity against fish pathogens, the antibiotic susceptibility, and the prevalence of virulence factors and detrimental enzymatic activities in 99 Lactic Acid Bacteria (LAB) (59 enterococci and 40 non-enterococci) isolated from aquatic animals regarded as human food. Results These LAB displayed a broad antimicrobial/bacteriocin activity against the main Gram-positive and Gram-negative fish pathogens. However, particular safety concerns based on antibiotic resistance and virulence factors were identified in the genus Enterococcus (86%) (Enterococcus faecalis, 100%; E. faecium, 79%). Antibiotic resistance was also found in the genera Weissella (60%), Pediococcus (44%), Lactobacillus (33%), b...