Probiotic Inoculation Research Articles

Development of an Apple Snack Enriched with Probiotic Lacticaseibacillus rhamnosus: Evaluation of the Refractance Window Drying Process on Cell Viability.

The objective of this study was to develop a dried apple snack enriched with probiotics, evaluate its viability using Refractance Window (RWTM) drying, and compare it with conventional hot air drying (CD) and freeze-drying (FD). Apple slices were impregnated with Lacticaseibacillus rhamnosus and dried at 45 °C using RWTM and CD and FD. Total polyphenol content (TPC), color (∆E*), texture, and viable cell count were measured, and samples were stored for 28 days at 4 °C. Vacuum impregnation allowed for a probiotic inoculation of 8.53 log CFU/gdb. Retention values of 6.30, 6.67, and 7.20 log CFU/gdb were observed for CD, RWTM, and FD, respectively; the population in CD, RWTM remained while FD showed a decrease of one order of magnitude during storage. Comparing RWTM with FD, ∆E* was not significantly different (p < 0.05) and RWTM presented lower hardness values and higher crispness than FD, but the RWTM-dried apple slices had the highest TPC retention (41.3%). Microstructural analysis showed that RWTM produced a smoother surface, facilitating uniform moisture diffusion and lower mass transfer resistance. The effective moisture diffusion coefficient was higher in RWTM than in CD, resulting in shorter drying times. As a consequence, RWTM produced dried apple snacks enriched with probiotics, with color and TPC retention comparable to FD.

Probiotics mitigate thermal stress- and pathogen-driven impacts on coral skeleton

Threats leading to a reduction in coral populations are apparent worldwide. Several different approaches have been tested to accelerate the adaptation of corals to a changing climate. Here, we evaluated the skeleton structure, crystal habit, and chemical changes of the coral Pocillopora damicornis in response to the pathogen (Vibrio coralliilyticus) and probiotic (Beneficial Microorganisms for Corals, BMCs) inoculation under ambient conditions (26 °C) and thermal stress (30 °C) during a 50-day mesocosm experiment. The skeletons were analyzed using microtomography, energy-dispersive x-ray spectroscopy (EDX/SEM), and densitometry to investigate the skeleto-physico-chemical micro-morphological changes in porosity, median pore-size diameter, crystal habit, Mg/Ca, Sr/Ca, the skeleton mineral density (g/cm2) and skeleton mineral content (g–2). The results indicate considerable changes in the coral skeleton caused by both temperature and microbial inoculation. Most importantly, lower density (to ∼ x̄ 0.5 g/cm2) and higher porosity (up to ∼ x̄ 47%) were correlated with inoculation of V. coralliilyticus and mitigated by probiotics. BMCs also substantially increased calcification, as evidenced by Mg/Ca in the skeleton of thermally stressed corals. At the micron scale, aragonite crystal fibbers precipitated during the experiments showed an acicular habit in thermally stressed and pathogen-inoculated corals kept at 30 °C. In contrast, a spherulitic habit, characteristic of high growth rates, was observed in corals inoculated with both BMCs and V. coralliilyticus. Our findings reveal that pathogen inoculation and thermal stress had notable impacts on coral skeleton properties, including porosity, density, and crystal morphology, in a short period of time, which highlights the potential impacts of shifts in climate warming and environmental quality. Interestingly, BMCs played a role in maintaining the properties of skeleton calcification.

Synergistic influence of probiotic and florfenicol on embryonic viability, performance, and multidrug-resistant Salmonella Enteritidis in broiler chickens

This study was conducted during the period of August 2021 to April 2022 and divided into two parts. The first part involved the isolation and characterization of Salmonella from 200 diseased broiler chickens collected from farms in Dakahlia Governorate, Egypt, with the detection of its antimicrobial susceptibility. The second experimental part involved in ovo inoculation of probiotics and florfenicol to evaluate their effects on hatchability, embryonic viability, growth performance traits and the control of multidrug-resistant Salmonella Enteritidis infections post hatching. The point prevalence of Salmonella in the internal organs of diseased chickens was 13% (26/200), including 6 serotypes: S. Enteritidis, S. Typhimurium, S. Santiago, S. Colindale, S. Takoradi and S. Daula. Multidrug resistance was found in 92% (24/26) of the isolated strains with a multiantibiotic resistance index of 0.33–0.88 and 24 antibiotic resistance patterns. The in ovo inoculation of probiotic with florfenicol showed significant improvement in the growth performance parameters compared with other groups and had the ability to prevent colonization of multidrug resistant S. Enteritidis in the majority of the experimental chicks, and the remaining chicks showed very low colonization, as detected by RT‒PCR. These findings suggested the application of in ovo inoculation techniques with both probiotics and florfenicol as a promising tool to control multidrug-resistant S. Enteritidis in poultry farms.

Evaluation of a Bacillus-based direct-fed microbial probiotic on in vitro rumen gas production and nutrient digestibility of different feedstuffs and total mixed rations.

We evaluated the effects of a Bacillus-based direct-fed microbial (DFM) on total in vitro gas production, dry matter (DM), neutral detergent fiber (NDF), and starch disappearance of different feedstuffs and total mixed rations (TMR) in three different experiments. In experiment 1, six single fiber-based feedstuffs were evaluated: alfalfa hay, buffalo grass, beet pulp, eragrostis hay, oat hay, and smutsvinger grass. Experimental treatments were control (with no probiotic inoculation; CON) or incubation of a probiotic mixture containing Bacillus licheniformis and B. subtilis (3.2 × 109 CFU/g; DFM). The calculation of DFM dose under in vitro conditions was based on the assumption of a rumen capacity of 70 liter and the dose of 3 g of the DFM mixture/head/d (9.6 × 109 CFU). Total in vitro gas production, DM, and NDF disappearance were evaluated at 24- and 48 h posttreatment incubation. Mean treatment effects were observed at 24- and 48 h gas production (P < 0.0001), as DFM incubation increased in vitro gas production by 5.0% and 6.5%, respectively. For nutrient digestibility, mean DM digestibility was increased at 48 h (P = 0.05), whereas mean NDF digestibility increased at both timepoints by incubating DFM in vitro (P ≤ 0.02). In experiment 2, nine commercial dairy TMR were collected and evaluated for the same variables and treatments described in experiment 1, with the additional analysis of starch digestibility at 7 h post in vitro incubation. The only difference was the concentration of the DFM included, being representative for a dosage of 8.8 × 109 CFU/head/d. In vitro gas production was increased only at 48 h due to DFM incubation (P = 0.05), whereas DM and NDF digestibility were improved at 24 and 48 h (P ≤ 0.02). No treatment effects were observed on in vitro starch digestibility (P = 0.31). In experiment 3, a combined analysis of DM and NDF digestibility was performed by using quality values (NDF and crude protein or CP) of 16 substrates. Regardless of CP and NDF levels of the substrates, DFM improved in vitro 24 and 48 h DM and NDF digestibility (P ≤ 0.03). In summary, incubating a Bacillus-based DFM (B. licheniformis and B. subtilis; BOVACILLUS) improved mean in vitro gas production, DM, and NDF digestibility of single feedstuffs and commercial dairy TMR, highlighting the potential of this combination of Bacillus spp. to improve nutrient utilization, mainly fiber.

Impact of early inoculation of probiotics to suckling piglets on postweaning diarrhoea – A challenge study with Enterotoxigenic E. Coli F18

Postweaning diarrhoea caused by Enterotoxigenic Escherichia coli (ETEC) is a threat to the pig industry. With an intensified focus on finding alternatives to the use of medical zinc oxide and antibiotics in newly weaned pigs, the objective of this study was to investigate the effect of early inoculation of probiotics to suckling piglets on subsequently ETEC faecal shedding and immune parameters in ETEC F18-challenged weaned piglets. Sixty pigs weaned on day 28 of age were assigned to three treatment groups: (i) Negative Control (non-challenged), (ii) Positive Control (challenged) and (iii) Probiotic (challenged and inoculated with a multi-species probiotic product during suckling). On days 1 and 2 postweaning, pigs in the Positive Control and Probiotic groups were challenged with 5 × 108 colony-forming unit ETEC F18/pig/day, whereas pigs in the Negative Control group were provided with NaCl. Growth and diarrhoea incidence were not significantly affected by ETEC challenge or probiotic administration. ETEC F18 shedding and C-reactive protein concentration in plasma were significantly lower in the Negative Control group, confirming a successful challenge model. Pigs in the Probiotic group had a significantly reduced number of pigs shedding ETEC F18 and STb toxin in faeces compared with the Positive Control group. Probiotic application did not significantly impact the concentration of C-reactive protein, haptoglobin and cytokines in plasma nor haematology numbers. In conclusion, weaned pigs administered with a multi-species probiotic product early in life had a more rapid response towards the pathogen challenge and a faster clearance of ETEC compared with the Positive Control group. Administration of a multi-species probiotic to newborn piglets may thus promote resilience in the newly weaned pig. However, further studies with pigs subjected to a more severe pathogen challenge are needed to confirm these results and to investigate the mechanism of action of the probiotic intervention.

Probiotics survival and betalains stability in purple pitaya (Hylocereus sp.) juice

The development of biofunctional non-dairy beverages with probiotics and prebiotics is a challenge and a frontier goal prompted by current consumers demand of health promoting foods, in particular from those with dietary restrictions related to dairy products. This study aimed to evaluate the survival of two commercially available probiotic strains, Lactobacillus rhamnosus LGG® and Lactobacillus paracasei (Lactobacillus casei 431), in a commercial sterile purple pitaya juice during storage at 5 °C. Additionally, the prebiotic effect of inulin (6.25 mg/ml) on the survival of L. rhamnosus, the most promising strain, was assessed. Physicochemical parameters such as betalain and soluble solids contents, pH, and color were also monitored during storage. Populations of L. rhamnosus and L. paracasei decreased from 11 to 7 log CFU/ml after 12 ± 1 and 0.93 ± 0.05 d, respectively. No changes on the individual betalain contents, as well as for total betalain concentrations (28 ± 3 μg/ml) were observed after probiotic inoculation nor during storage for all experimental trials and regardless of the strain evaluated (P < 0.05). Moreover, the addition of inulin did not significantly (P > 0.05) influence the survival kinetics of L. rhamnosus in the juice during storage and the juice color, pH and soluble solids content remained unaffected as well. Purple pitaya juice represents therefore a promising non-dairy food matrix for probiotics delivery.

Effects of a Bacillus-based direct-fed microbial on in vitro nutrient digestibility of forage and high-starch concentrate substrates.

Two experiments evaluated the effects of a Bacillus-based direct-fed microbial (DFM) on in vitro dry matter (DM) and neutral detergent fiber (NDF; experiment 1) and starch (experiment 2) digestibility of a variety of ruminant feedstuffs. In experiment 1, 10 forage sources were evaluated: ryegrass, alfalfa hay, leucaena, corn silage, spinifex, buffel grass, flinders grass, Mitchell grass, Rhodes grass hay, and Queensland bluegrass. Experimental treatments were control (forages with no probiotic inoculation; CON) and forage sources inoculated with a mixture containing Bacillus licheniformis and Bacillus subtilis (3.2 × 109 CFU per g; DFM). In vitro DM and NDF digestibility were evaluated at 24- and 48-h post-treatment inoculation. Treatment × hour interactions were noted for IVDMD (in vitro dry matter digestibility) and IVNDFD (in vitro neutral detergent fibre digestibility) (P ≤ 0.05). More specifically, DFM inoculation increased (P ≤ 0.03) IVDMD at 24 h in four forages and increased 48-h IVDMD (P ≤ 0.02) in alfalfa hay, ryegrass, leucaena, and Mitchell grass hay, but opposite results were observed for Queensland bluegrass (P < 0.01). A 24- and 48-h IVNDFD increased following DFM inoculation (P ≤ 0.02) in five forage sources, but reduced for Queensland bluegrass (P < 0.01). When the forages were classified according to their quality, main treatment effects were detected for IVDMD (P ≤ 0.02) and IVNDFD (P < 0.01). In experiment 2, five common cereal grains were evaluated—high-density barley (82 g/100 mL), low-density barley (69 g/100 mL), corn, sorghum, and wheat—under the same treatments as in experiment 1. In vitro starch digestibility (IVSD) was evaluated at 6- and 12-h following treatment inoculation. Treatment × hour interactions were observed for starch digestibility in three out of five concentrate sources (P ≤ 0.001). Inoculation of DFM yielded greater 24-h starch digestibility for high-, low-density barley, and wheat (P ≤ 0.02), but also greater at 48 h in wheat (P < 0.0001). Moreover, mean starch digestibility improved for corn and sorghum inoculated with DFM (P < 0.01). Using a Bacillus-based DFM (B. licheniformis and B. subtilis) improved the mean in vitro DM and NDF digestibility of different forage sources of varying qualities (based on crude protein content). Similarly, IVSD was also greater following DFM inoculation, highlighting the potential of this probiotic to improve nutrient digestibility and utilization in the beef and dairy cattle herd.

Characteristics of functional ice cream produced with probiotic Saccharomyces boulardii in combination with Lactobacillus rhamnosus GG

Ice creams have big potential to be used as probiotic food carrier. In this respect, this study aimed to produce ice creams with addition of Saccharomyces boulardii and Lactobacillus rhamnosus GG. Probiotic strains were added to the ice cream mixes before and after aging steps of ice cream production and were inoculated as single or mix cultures. Probiotic ice cream products were then characterized and the survival of the probiotic strains were determined during storage period. Both probiotic strains successfully survived during storage period and their numbers were above 6.18 log CFU/g at final date of storage. The rheological parameters of ice cream products improved with the co-inoculation of S. boulardii and L. rhamnosus both before and after aging step. Importantly the aroma profile of ice cream samples altered with probiotic inoculation and ethanol was the major volatile compound in ice cream samples produced with S. boulardii added before aging step. Inoculation of S. boulardii and L. rhamnosus as single cultures also resulted in formation of distinct aroma compounds in ice cream products. The highest overall acceptability score in sensorial evaluation of ice cream products was obtained from the ice cream products produced with inoculation of probiotics after aging step.

In ovo inoculation of probiotics for broiler chickens: Systematic review and meta-analysis

The objective of this study was to evaluate the effect of probiotic inoculation in fertilized eggs on hatching and posthatching characteristics of broilers. The search for scientific articles was conducted in December 2020 in eight databases (Embase, Google Scholar, SciELO, Science Direct, Scopus, Periodicos Capes, PubMed, and Web of Science) using the following combination of keywords: ((probiotic OR synbiotic) AND "in ovo"). Only articles that evaluated the effect of in ovo inoculation of probiotics, combined or not with other compounds, on hatching parameters (number of hatched eggs and hatching weight) and/or posthatching parameters (performance and carcass yield) in broilers were included. The meta-analysis was performed using the random-effects model to calculate the confidence interval (CI) of the differences between the group inoculated with probiotics and the control (vehicle inoculation). The degree of heterogeneity was assessed by meta-regression, and publication bias was assessed by funnel plots. Wide heterogeneity was observed (P < 0.01) in all analyzes. The overall analysis showed that probiotic inoculation did not influence (P > 0.05) number of hatched eggs, hatching weight, or feed intake after hatching but increased (P < 0.01) weight gain and reduced (P < 0.05) feed conversion ratio. Subgroup analysis showed that, for performance, the Bifidobacterium as probiotic brought the best results, while the yolk was best inoculation site. We conclude that probiotics can improve posthatching performance without influencing hatching parameters, depending on the type of microorganism, inoculation site, and broiler strain used.

Effects of in ovo probiotic administration on the incidence of avian pathogenic Escherichia coli in broilers and an evaluation on its virulence and antimicrobial resistance properties

Avian pathogenic Escherichia coli (APEC) causes colibacillosis in poultry, which has been traditionally controlled by the prophylactic in-feed supplementation of antibiotics. However, antibiotics are being removed from poultry diets owing to the emergence of multidrug-resistant (MDR) bacteria. Therefore, alternatives to control APEC are required. This study aimed to evaluate the effects of in ovo inoculation of probiotics on the incidence of APEC in broilers and evaluate the virulence and antimicrobial resistance properties of the APEC isolates. On embryonic day 18, 4 in ovo treatments (T) were applied: T1 (Marek's vaccine [MV]), T2 (MV and Lactobacillus animalis), T3 (MV and Lactobacillus reuteri), and T4 (MV and Lactobacillus rhamnosus). A total of 180 male broilers per treatment were randomly placed in 10 pens. The heart, liver, spleen, and yolk sac were collected on day 0, 14, 28, and 42. Presumptive E. coli isolates were confirmed by real-time PCR. The positive isolates were screened for the APEC-related genes (iroN, ompT, hlyF, iss, and iutA), and E. coli isolates containing one or more of these genes were identified as APEC-like strains. A total of 144 APEC-like isolates were isolated from 548 organ samples. No differences (P > 0.05) among treatments were observed for the incidence of APEC-like strains in all organs when averaged over sampling days. However, when averaged over treatments, the incidence in the heart, liver, and yolk sac was different among sampling days; a significant increase was observed in these organs on day 14 compared with day 0. Twenty-five antimicrobial resistance genes were evaluated for all APEC-like isolates, and 92.4% of the isolates carried at least one antimicrobial resistance gene. Thirty-seven isolates were then selected for antimicrobial susceptibility testing; MDR strains accounted for 37.8% of the isolates. In conclusion, the in ovo inoculation of a single probiotic strain did not confer protection against APEC strains in broilers. The high prevalence of MDR isolates indicates that further research on antibiotic alternatives is required to prevent APEC infections in broilers.

In ovo inoculation of an Enterococcus faecium–based product to enhance broiler hatchability, live performance, and intestinal morphology

Previous studies have suggested the use of probiotics, as alternative to antibiotics, to enhance broiler performance. The administration of probiotics in feed has been widely explored; however, few studies have evaluated the in ovo inoculation of probiotics. Therefore, the objective was to evaluate the impact of in ovo inoculation of different concentrations of GalliPro Hatch (GH), an Enterococcus faecium–based probiotic, on hatchability, live performance, and gastrointestinal parameters. Ross x Ross 708 fertile eggs were incubated, and on day 18, injected with the following treatments: 1) 50 μL of Marek's vaccine (MV), 2) MV and 1.4 × 105 cfu GH/50 μL, 3) MV and 1.4 × 106 cfu GH/50 μL, 4) MV and 1.4 × 107 cfu GH/50 μL. On the day of hatch, chicks were weighed, feather sexed, and hatch residue was analyzed. Male birds (640) were randomly assigned to 40 floor pens. On day 0, 7, 14, and 21 of the grow-out phase, performance data were collected. One bird from each pen was used to obtain yolk weight and intestinal segment weight and length. Hatchability was not impacted by any GH treatment (P = 0.58). On day 0, yolk weight was lower for all treatments than for MV alone. On day 0 to 7, feed intake was lower for 105 and 107 GH; the feed conversion ratio (FCR) was lower for all treatments than for MV alone (P = 0.05; P = 0.01, respectively). From day 14 to 21, the 107 GH treatment had higher BW gain (P = 0.05). For day 0 to 21, 107 GH had a lower FCR than MV alone (P = 0.03). On day 0, all GH treatments resulted in heavier tissues and longer jejunum, ileum, and ceca lengths than MV alone (P < 0.05). Spleen weight was higher for 105 and 107 GH than for MV alone. In conclusion, GH does not impact hatchability, and some concentrations improved live performance through the first 21 d of the grow-out phase. These improvements could result from the increased yolk absorption and improved intestinal and spleen morphology seen in this study.

The Inoculation of Probiotics In Vivo Is a Challenge: Strategies to Improve Their Survival, to Avoid Unpleasant Changes, or to Enhance Their Performances in Beverages

The inoculation of probiotics in beverages (probiotication) requires special technologies, as probiotic microorganisms can experience stress during food processing (acid, cold, drying, starvation, oxidative, and osmotic stresses) and gastrointestinal transit. Survival to harsh conditions is an essential prerequisite for probiotic bacteria before reaching the target site where they can exert their health promoting effects, but several probiotics show a poor resistance to technological processes, limiting their use to a restricted number of food products. Therefore, this paper offers a short overview of the ways to improve bacterial resistance: by inducing a phenotypic modification (adaptation) or by surrounding bacteria through a physical protection (microencapsulation). A second topic briefly addressed is genetic manipulation, while the last section addresses the control of metabolism by attenuation through physical treatments to design new kinds of food.

The potential for inoculating Lactobacillus animalis and Enterococcus faecium alone or in combination using commercial in ovo technology without negatively impacting hatch and post-hatch performance

The poultry industry has recently undergone transitions into antibiotic free production, and viable antibiotic alternatives, such as probiotics, are necessary. Through in ovo probiotic inoculation, beneficial microflora development in the gastrointestinal tract may occur prior to hatch without negatively impacting chick performance. Therefore, the objective of the present study was to observe the impacts of the injection of probiotic bacteria individually or combined into fertile broiler hatching eggs on hatch and live performance characteristics. A total of 2,080 fertile broiler hatching eggs were obtained from a commercial source. On day 18 of incubation, 4 in ovo injected treatments were applied: 1.) Marek's Disease (HVT) vaccination, 2.) L. animalis (∼106 cfu/50μl), 3.) E. faecium (∼106 cfu/50μl), and 4.) L. animalis + E. faecium (∼106 cfu & ∼106 cfu/50μl each). On day of hatch, hatchability and hatch residue data were recorded. A portion of male chicks from each treatment were placed in a grow-out facility for a 21 d grow-out (18 chicks/pen × 10 pens/treatment = 720 male chicks) with a corn and soy bean meal-based diet without antibiotics or antibiotic alternatives. Performance data and gastrointestinal samples were collected on days 0, 7, 14, and 21. Results indicated no differences in all hatch parameters between treatments (P > 0.05) except for % pipped, where the L. animalis treatment had lower % pipped eggs compared to the HVT control and E. faecium treatments (P = 0.04). No differences were observed in body weight gain or mortality (P > 0.05). Probiotic treatments altered gastrointestinal tissue length, weight, and pH. This resulted in all in ovo injected probiotic treatments increasing feed conversion ratio (FCR) from days 7 to 14 as compared to the control (P = 0.01). Differences in FCR were not observed in any other week of data collection (days 0 to 7, 14 to 21, or 0 to 21; P > 0.05). Although probiotics altered live performance from days 7 to 14, these data suggest that in ovo inoculations of L. animalis and E. faecium in combination are viable probiotic administration practices that potentially improve hatch characteristics and gastrointestinal tract development.

Effect of in Ovo Inoculation of Bifidobacterium spp. on Growth Performance, Thyroid Activity, Ileum Histomorphometry, and Microbial Enumeration of Broilers.

Early colonization by beneficial bacteria stimulates the function and development of the digestive tract, on which the performance and vitality of broilers rely. This experiment evaluated the effects of in ovo injection of bifidobacteria on the performance, thyroid activity, bacterial enumeration, and ileal histomorphometry of broiler chickens. A total of 360 fertile eggs were inoculated into the yolk sac, on day 17 of embryogenesis, and allocated to six experimental groups: negative control (not injected, G1), positive control (injected with sterilized distilled water, G2), and groups inoculated with 2 × 108 CFU of Bifidobacterium bifidum, G3; B. animalis, G4; B. longum, G5; or B. infantis, G6. The results revealed an increase (P < 0.01) in body weight and weight gain in all treated groups increases of at least 5.38 and 8.27%, respectively, compared with the control. Feed consumption was not affected during all experimental periods, while the feed conversion ratio (FCR) was enhanced (P < 0.01) only for the overall experimental period (1-28 days of age). The G3 birds recorded the lowest FCR (1.38), while the highest was observed in G1 birds (1.57). Serum concentrations of thyroxin and triiodothyronine were elevated (P < 0.05) with probiotic inoculation. The antioxidant status and immune response of bifidobacteria injected birds were improved; the serum contents of superoxide dismutase and immunoglobulins Y, M, and A were increased (P < 0.05 and P < 0.01), while the malondialdehyde content was decreased (P < 0.01). Ileal architecture was improved in the bifidobacteria treated groups; the highest values of villus height and the villus height/crypt depth ratio were recorded in G3 (936.6 and 11.80) compared with those of G1 (537.1 and 6.93). Moreover, ileal lactic acid bacteria and Bifidobacterium spp. counts increased by at least 10.64 and 51.75%, while total coliform and bacterial counts reduced by at least 15.46 and 15.18%, respectively, compared with those of the control. In conclusion, all tested strains of bifidobacteria enhanced broiler growth performance, ileal function, and thyroid hormone metabolism without obvious differences among them.

Short‐term effect of the inoculation of probiotics in mature bioflocs: Water quality parameters and abundance of heterotrophic and ammonia‐oxidizing bacteria

The aim of this study was to evaluate the short-term effect of probiotic inoculation on the abundance of heterotrophic and ammonia-oxidizing bacteria in mature biofloc, as well as on total suspended solids (TSS), chlorophyll-a and nitrogenous compounds in water. A completely randomized design consisting of five treatments (three commercial probiotics, one native consortia and one control) was performed. At the beginning of the experiment (day 1), each treatment was inoculated with the respective probiotic: PondPlus® (PP), Efinol® PT (EF) and Epicin® ponds (EP), native consortia UE, whereas the control was not inoculated. Water parameters and bacterial abundance were evaluated at 0, 2, 4, 6 and 8 days. The addition of probiotics, either native or commercial, did not show any significant effect on the TSS, Chl-a and colony-forming unit (CFU) of heterotrophic bacteria when they were added to the systems containing mature biofloc. A significant increase in ammonium oxidizing bacteria was registered with the probiotics PP and EP, although the levels of total ammonia nitrogen, NO3-N and NO2-N were statistically similar among all treatments. Modifications on most of the parameters measured were associated with the factor of time, rather than the inclusion of probiotics. Results suggest that the bacterial conglomerates in mature stage contain well-established bacterial communities that are difficult to be affected by the addition of probiotics.

Patulin removal from synbiotic apple juice using Lactobacillus plantarum ATCC 8014.

The aim of this study was to evaluate the elimination of patulin (PAT) by Lactobacillus plantarum ATCC 8014 from artificially contaminated apple juice and its dependence on prebiotic, citric acid and ascorbic acid content. A central composite design was used for studying each of the three factors at five levels to find the optimum concentrations. The results showed that inserting 2·3% (w/v) fructooligosaccharide, 213mgl-1 ascorbic acid and 1·4gl-1 citric acid to apple juice with inoculating 3·6×1011 CFU per ml, L. plantarum improved the efficiency of PAT removal to 95·91% during 6weeks cold storage. SDS-PAGE of cell surface proteins of probiotics revealed that surface layer proteins have an important role in PAT removal from apple juice. No significant difference was observed in the flavour and colour of the optimized synbiotic apple juice and in the control sample until 3weeks of cold storage. Lactobacillus plantarum ATCC 8014 is capable of PAT removal from artificially contaminated synbiotic apple juice. Synbiotic apple juice artificially contaminated with PAT will be safe for consumers after the first day of probiotic inoculation; and surface layer proteins of probiotic cells are responsible for PAT removal.

Inclusion of prebiotics on the viability of a commercial Lactobacillus casei subsp. rhamnosus culture in a tropical fruit beverage.

The viability of probiotics in the development of functional tropical fruit beverages is a technological challenge that may benefit from the addition of prebiotics, due to the synergistic (synbiotic) interaction. This study evaluated the viability of a commercial probiotic (Lactobacillus casei) in a blended red fruit beverage (RFB; 20% strawberry, 10% blackberry and 5% papaya), enriched with three separate prebiotics: inulin (IN), fructooligosaccharides (FOS) and galactooligosaccharides, added at 1 and 5%. The consumer preference for the beverages was also examined. The inoculum was produced in MRS broth supplemented with 10% RFB, which reached the exponential phase (9.96 log CFUmL-1) after incubation at 37°C for 24h. In search of the probiotic strain's adaptation and viability in the presence of the different prebiotics (measured by optical density, OD600 nm), the prebiotics were added to MRS broth at 1 and 5%. Since 1% IN (OD = 3.99 ± 0.36) and 1% FOS (OD = 3.48 ± 0.28) were the most significant, these treatments, without inoculation of probiotics, were assessed by the sensory panel. Although neither RFB was significantly preferred the RFB with 1% IN received the greatest number of responses (n = 33/60). Its effect on the viability of L. casei inoculated in the RFB was monitored by the growth kinetics at 37°C for 50h. The findings indicated that fortification with 1% IN could have a possible protective effect on the stability of L. casei in RFBs, highlighting the use of tropical fruits as potential carriers of probiotics.

Fermented milk beverage: formulation and process

ABSTRACT: Dairy products are an adequate alternative as functional foods since they present excellent nutritional value, have wide acceptance with consumers, and are potential matrices for inoculation of probiotic microorganisms. The present study aimed to develop a fermented milk beverage with fruits and a probiotic culture included, to evaluate the acceptance of this product among consumers to define the ideal formulation, and to perform Lactobacillus acidophilus counting in order to characterize the product as a probiotic food. For the formulation of the beverage, milk, whey, sugar, and pasteurized orange and papaya pulps were used as main ingredients. The beverage formulation consisted of mixing the fruit pulps with a dairy substrate of milk and whey which was previously fermented using a mixed commercial probiotic ABT-5 culture. Six formulations were tested. The milk substrate was maintained at 70%. These formulations consisted of 50% of mineral water and 50% of skimmed milk with a concentration of probiotics of 0.5g/L. Sugar concentration varied between 40g, 70g, or 100g. As for flavoring, the juice was composed of 180mL of papaya and 120mL of orange, or 150mL of papaya and 150mL of orange. Beverages were prepared and then stored at 5±1°C. These products were evaluated after 24 hours of preparation/storage by 90 tasters (consumers) for acceptance and sensory characterization. Data were analyzed by ANOVA. The differences between samples were evaluated using the Tukey test at a significance level of 5%. Acceptance of the beverage samples among tasters varied from 5.1 to 6.9 and ranged between the hedonistic terms I neither liked nor disliked it and somewhat enjoyed it in a 9-point hedonic scale. The enumeration of Lactobacillus acidophilus LA-5 ranged from 7.83 to 8.09log CFU/mL in the first week of refrigerated storage. Our results suggested that the beverages analyzed were suitable for inoculation of probiotics and met the legislation requirements. Therefore, these beverages should be classified as probiotic products. Samples with 10% sugar were the least preferred by the study participants whereas beverages containing 40g, 70g and 180mL of papaya juice and 120mL of orange juice and the sample with 70g of sugar, 150mL of papaya juice, and 150mL of orange juice did not differ significantly from each other in terms of acceptance and were; therefore, the 3 formulations that received the highest means of acceptance by consumers.

Viability of Lactobacillus plantarum on Fresh-Cut Chitosan and Alginate-Coated Apple and Melon Pieces.

There is an increasing trend toward foods with probiotics; the awareness of healthy diet and wellbeing is the leading cause of this increase. As a result, food producers and stakeholders require new probiotic products. The increased incidence of lactose intolerance and the new lifestyles (vegan and vegetarian styles) have led to a renewed interest in non-dairy probiotic carriers. The use of biopolymeric matrices to develop active food packaging carrying probiotics has been studied and proposed as an alternative method to design new solutions.The main topic of this paper was the design of fresh-cut fruits (apples and melons) as carriers for a promising Lactobacillus plantarum; fruit pieces were coated with either alginate or chitosan. Apple (Granny Smith) and melon pieces (Cucumis melo, var. Cantalupensis) were preliminary treated with an anti-browning solution (citric and ascorbic acids). Then, fruit pieces were dipped in a solution containing L. plantarum c19 (9 log cfu/ml) and coated with alginate or chitosan. Samples without probiotic and/or coatings were used as controls.All samples were stored at 4°C for 14 days under air or modified atmosphere (65% N2, 30% CO2, and 5% O2); the following analyses were done: pH, color, O2, and CO2 in the head space, microbiology (mesophilic bacteria, lactic acid bacteria, yeasts, and molds).The most important results can be summarized as follows: (a) Alginate coating showed better performances than chitosan-coating, as it did not affect the viability of L. plantarum. (b) The inoculation of probiotics in the controls negatively affected the color, but the coating was able to counteract this effect. This paper supports the combination of edible coatings and probiotic as a promising way to design new fruit-based functional foods; further investigations are required to study the effect of this combination on the sensory scores.

Use of indigenous beneficial bacteria (Lactobacillus spp.) as probiotics in shrimp (Penaeus monodon) aquaculture

The present study was conducted to study in-vitro the antagonistic effect of Lactobacillus spp. against the pathogenic bacterial vibrio harveyi on shrimp. For this purpose, shrimp samples were collected from three different Ghers at Batiaghata upazilla, Khulna. Gills and intestines were taken out from the samples to identify the load of Lactobacillus spp. and Vibrio spp. The results revealed that the load of Lactobacillus spp. was found more than Vibrio spp. both in gills and intestines; the gills also contained higher load of Vibrio spp. than in the intestines. V. harveyi was separated from the isolated Vibrio spp. with different types of biochemical tests: Gram stain, Motility test, Indole test, VP test, MR test, Arginine dihydrolase, Salt tolerance test, growth at different temperature ranges and colony color on TCBS agar media. The isolated V. harveyi was subjected for in-vitro test. In in-vitro challenge test, the potential antagonistic effect of Lactobacillus spp. against V. harveyi was gradually obtained at 0, 4th, 8th, 12th hour of treatments. Interesting finding was that, with the time, the load of V. harveyi was reduced gradually and the lowest load was obtained after 12 hours of probiotic inoculation. The present study revealed an excellent in-vitro antagonistic probiotic effect of Lactobacillus spp. on V. harveyi. Therefore the result suggested that probiotic treatment might be an effective alternative to the use of antibiotics in treatments of bacterial diseases in shrimp aquaculture.Res. Agric., Livest. Fish.5(1): 127-135, April 2018