Probiotic Potential and Immunomodulatory Properties in Enterococcus Faecium GMB24 and Enterococcus Hirae SMB16 Isolated from Goat and Sheep Milk

Author(s):  
Kamni Rajput ◽  
Ramesh Chandra Dubey ◽  
Ashwani Kumar Jangra

Abstract Probiotic attributes of lactic acid bacteria isolated from goat and sheep milk samples were analysed by culturing them on MRS (de Man, Rogosa and Sharpe) agar media. The most potential isolates, GMB24 and SMB16, were identified by biochemical tests, which were further identified as Enterococcus faecium GMB24 and Enterococcus hirae SMB16 by 16S rRNA gene sequencing approach. The probiotic potential of the GMB24 and SMB16 were possessed for probiotics attributes including antimicrobial activity against five pathogenic bacteria viz., Escherichia coli (MTCC118), Staphylococcus aureus (MTCC7443), Pseudomonas aeruginosa (MTCC424), Listeria monocytogens (MTCC657) and Salmonella typhimurium (MTCC733), and antibiotic susceptibility test. These bacterial isolates had ability to tolerate different concentrations of acid, bile and phenol resistance. Further, immunomodulating activities of potential probiotic bacteria were tested on neutrophil adhesion test, haemagglutinating antibody titer and delayed type hypersensitivity. The data from these experiments were used for the principal component analysis (PCA) for assessing survivability of isolates at different factors. The probiotic bacterial isolates showed good response for the enhanced antibody production and delayed type hypersensitivity (DTH) activity. Probiotic isolates E. faecium GMB24 and E. hirae SMB16, at 109 cells/ml doses per day, increased the neutrophil adhesion, haemagglutinating antibody titer and DTH in comparison to the untreated control group. These differences were statistically significant (P < 0.05). The isolates showed negative test for hemolytic and gelatinase activities and hence were considered safe. E. faecium GMB24 and E. hirae SMB16 possessed an excellent probiotic potential and had a strong potential immune-stimulant activity.

Author(s):  
Joshua Nfambi ◽  
Godfrey S. Bbosa ◽  
Lawrence Fred Sembajwe ◽  
James Gakunga ◽  
Josephine N. Kasolo

AbstractGlobally,An experimental laboratory-based study was done following standard methods and procedures. Nine experimental groups (I, II, III, IV, V, VI, VII, VIII, IX) each comprising of six animals were used. Group I received normal saline. Groups II to IX received 200 mg/kg bwt cyclophosphamide at the beginning of the study. Group III received 50 mg/kg bwt of an immunostimulatory drug levamisole. Groups IV to IX were dosed daily for 14 days with extract at doses of 250, 500, and 1000 mg/kg bwt, respectively, using an intragastric tube. Complete blood count (CBC), delayed-type hypersensitivity reaction (DTH), neutrophil adhesion test, and hemagglutination antibody titer were determined using standard methods and procedures. Statistical analysis was performed using GraphPad prism 5.0a Software.There was an increment in WBC, lymphocyte, and neutrophil counts at a dose of 1000 mg/kg bwt similar to the levamisole-positive control group. The neutrophil adhesion was statistically significant (p≤0.05) for treatment groups that received 1000 mg/kg bwt (29.94%) and 500 mg/kg bwt at 17.28%. The mean percentage increment in footpad thickness was highest (26.9%) after 8 h of injection of antigen in the footpad of rats dosed 500 mg/kg bwt and this later reduced to 25.6% after 24 h. There was a dose-dependent increment in the mean hemagglutination antibody titer to sheep red blood cells (SRBC) from 10.73±0.57 HA units/μL for the 250 mg/kg bwt to 26.22±1.70 HA units/μL for the 1000 mg/kg bwt.Methanolic leaf extract of


2021 ◽  
pp. 464-473
Author(s):  
Dicki Bakhtiar Purkon

Immunostimulants are compounds that can stimulate an immune response by increasing the activity of non-specific and specific components of the immune system (humoral and cellular) against certain infections and diseases. The liverwort plant species Marchantia paleacea Bertol. has long been used as a source of nutrition and empirical medicine. However, scientifically there is still not much research data on immunomodulators in these plants. This study aims to determine the activity of immunomodulators in the ethanol extract of the herb Marchantia paleacea Bertol. in male mice of BALB/c strain. Bioactive compounds from this plant were extracted by maceration method using 96% ethanol. Extract characterization and phytochemical screening were determined according to WHO guidelines and standard procedures from previous studies. The immunomodulatory activity of the extract was tested by carbon clearance method and lymphoid organ index (non-specific responses), primary and secondary antibody titer tests (humoral specific responses), IL-2 cytokine levels and IFN-ɣ from serum secondary antibodies and delayed-type hypersensitivity reaction/DTH (cellular specific response). The results of qualitative phytochemical screening contained flavonoid compounds, saponins, phenolics, tannins and steroids/triterpenoids. The results of the non-specific immune response immunomodulator test showed that the dose of 52 mg/kg bw had the largest phagocytic index of 1.52 which included strong immunostimulation (K > 1.5) and the organ spleen index of 0.55 ± 0.11 which increased significantly compared to the control (p<0.05). The data on the acquisition of specific immune responses in the primary and secondary antibody titer test in the three test extracts resulted in increased titer levels compared to the control and at a dose of 52 mg/kg bw could significantly increase the levels of IL-2 cytokines in the control group (p<0,05). Meanwhile, in the DTH test, doses of 13 and 26 mg/kg bw could significantly increase the thickness of the soles of mice compared to controls (p<0.05).


2019 ◽  
Vol 7 (3) ◽  
pp. 114-119
Author(s):  
Dhanapal Venkatachalam l

Objective: A large number of plants and their isolated constituents have been shown to have potential immunity.In this review focusing on the immuno-modulatory activity of Ashwagandha (Withania somnifera) belonging to Solanaceae family and Neem (Azadirachta indica) belonging to Meliacea family. Methods: The Aswagandha churna was evaluated immunomodulatory activity byusing Albino wester rats and measured Delayed–type hypersensitivity response (DTH) (footpad swelling) and Percent neutrophil adhesion. The immuno-stimulatory effect of aswagandha root extract (WSE) evaluated by dexamethasoneinduced immunosuppressed mice in-vitro model.  The immunomodulatory effectof Neem leaves infusion was evaluated by using broiler chicks, measuring theantibody titer determination against Infectious Bursal Disease (IBD) using ELISA Kit techniques.The effects of Neem on some hematological parameters were measured the Estimation of Packed Cell Volume (PCV) and total leucocyte count (TLC). Results: The results obtained from this study, Ashwagandha churna when orally administered the neutrophil adhesion (p<0.05) was significantly increased with the dose of 300mg kg -1day-1 when compared with untreated control indicating possible immune-stimulant effect, and the DTH response, which is a direct correlate of cell mediated immunity (CMI), was significantly increased at a doses of 200 and 300mg kg-1 day - In vitro immuno-stimulatory activity  of  Aswagandha  root extract remarkably enhanced lymphocytes proliferation compared with the negative control (untreated cells). Both 2.5 μl and 5 μl of WSE (35 mg/ml) revealed significant (P < 0.001) stimulation of lymphocytes proliferation by approximately 322% and 403% respectively compared with the untreated cells. The immunomodulatory action of Neem leaves infusion; the mean antibody titer against IBD was higher for group C than all other groups. The effects of neem on some Hematological parameters of Packed cell volume of the control group was significantly lower than that of the treated groups at p<0.001, but with that of C2 at p<0.05. Conclusions: The current review on Immunomodulatory herbs shows great potential of herbs to cure or prevent certain diseases, which are beyond the reach of allopathic medicines in certain cases.    


2019 ◽  
Author(s):  
Ismail Marzuki ◽  
Andi Emelda

The aim of this study was to investigate the immunomodulatory effect of ethyl acetate of extract of Passiflora foetida L leaves (EAEPL) on hypersensitivity response and antibody titer of immune cells. Hypersensitivity response test evaluated the swelling of the feet of mice, while antibody titer test evaluated the hemagglutination. Examination of antibody titer was performed in 5 groups of 25 wistar rats. All the test animals, except control group, induced by SRBC on the 3rd day intraperitoneally. The extract was given for 7 days. The hypersensitivity response test was performed in 5 groups of 25 mice (Balb/c). The groups, except control group, were given antigen (SRBC) on the 3rd day intraperitoneally and on the 7th day were administered intraplantarly. The thickness of the soles of the feet was measured using a plethysmometer before and after injecting the antigen at T4, T24 and T48. The data were processed statistically by Kruskal-Wallis then advanced test by Mann-Whitney. The group of negative control against the groups of EAEPL was significantly different (p&lt;0.05). The antibody titers of the groups given the extract were higher than the antigen-induced group without the administration of the extract. The results show that the groups of EAEPL can increase antibody secretion. The test of hypersensitivity response showed that the group of negative control to EAEPL groups showed significantly different (p&lt;0.05). The results showed that the swelling of the feet of mice decrease at T48. This research concludes that ethyl acetate extract of EAEPL has the immunostimulatory effect.


1999 ◽  
Vol 43 (5) ◽  
pp. 1137-1143 ◽  
Author(s):  
Beatriz Robredo ◽  
Kavindra V. Singh ◽  
Fernando Baquero ◽  
Barbara E. Murray ◽  
Carmen Torres

ABSTRACT Fifteen newborn chickens were isolated in separate cages after 1 month of living together, divided into three groups, and challenged for 5 weeks with seed food which either was supplemented with avoparcin (10 mg/kg of animal food) or tylosin (40 mg/kg) or was nonsupplemented. At 9 weeks of age and after the 5-week challenge, all chickens received nonsupplemented feed for 4 additional weeks. At 4, 9, and 13 weeks of life, feces were collected and inoculated on M-Enterococcusagar plates with and without vancomycin (4 μg/ml).vanA-containing Enterococcus hirae was isolated from 11 of 15 chickens before antibiotic challenge, without detection of vancomycin-resistant Enterococcus faecium. At 9 weeks of age and after the 5-week avoparcin challenge, vanA E. hiraestrains were no longer detected, but five of five chickens now hadvanA E. faecium. At a lower frequency, vanA E. faecium had also displaced vanA E. hirae in both the tylosin group (one of four chickens) and the control group (two of five chickens). One month after avoparcin discontinuation, the number of chickens colonized with vanA E. faecium decreased from five to one. All vanA-containing E. hirae strains detected in the first month of life and most of thevanA-containing E. faecium strains detected in the second month of life showed identical ApaI andSmaI restriction patterns, respectively, when analyzed by pulsed-field gel electrophoresis. All vanA E. hiraeisolates transferred glycopeptide and macrolide resistance toEnterococcus faecalis JH2-2 in vitro; the level of glycopeptide resistance was higher in the transconjugants than in the donor E. hirae strains. These data suggest that E. hirae may be a significant source of vanAdeterminants with the potential of transfer to other enterococcal species from humans or animals.


2013 ◽  
Vol 16 (4) ◽  
pp. 619-627 ◽  
Author(s):  
M. Pogány Simonová ◽  
A. Lauková ◽  
L. Chrastinová ◽  
I. Plachá ◽  
V. Strompfová ◽  
...  

Abstract The effect of Enterococcus faecium CCM7420 (EF) - enterocin-producing and probiotic strain of rabbit origin, Eleutherococcus senticosus extract (ES) and their combination (ES+EF) was determined on selected bacteria in faeces and caecum content, leukocytes phagocytosis, blood biochemistry and growth performance. Ninety-six weaned rabbits were divided into 3 experimental (ES, EF, ES+EF) and control group (CG). The rabbits in the groups ES and EF+ES were fed commercial diet enriched with E. senticosus extract (30 g/100 kg feed), rabbits in groups EF and CG were fed untreated diet. The rabbits in the EF and ES+EF groups were administered with an overnight culture of E. faecium CCM7420 strain (500 μl/animal/day into water, 109 CFU/ml). The treatment period lasted 21 days. The microbiological examinations in faecal samples confirmed the presence of E. faecium CCM7420 strain. In groups EF and ES+EF, the reduction of faecal coliforms, Pseudomonas-like sp., Clostridium-like sp. and S. aureus was recorded. Leucocyte phagocytosis significantly increased in all experimental groups (P<0.0001) compared to CG. The lowest GPx values were measured in the ES+EF group. Higher total protein, triglycerides and calcium concentrations were detected in experimental groups compared to CG. The cholesterol concentration decreased in the ES group. The highest average daily gain was recorded in EF group; in ES+EF the better feed conversion ratio and no mortality was recorded. These results indicated that the dietary supplementation with the E. faecium CCM7420 and E. senticosus extract stimulate the leukocytes phagocytosis and reduces the potential pathogens in rabbits digestive tract without oxidative stress and improve the growth performance.


Author(s):  
Andrea Lauková ◽  
Anna Kandričáková ◽  
Eva Bino

This study investigated eight types of Slovak dry fermented meat products (salami and sausages) that are available on the market and were produced by three different producers in different regions of Slovakia. The total counts of enterococci in these products ranged from 2.0 up to 6.0 cfu/g (log10). Three species were identified among the 15 selected enterococcal strains; Enterococcus faecium (8 strains), Enterococcus faecalis (3) and Enterococcus hirae (4). They were hemolysis-negative (γ-hemolysis) with a biofilm-forming ability, which was evaluated as low-grade biofilm formation, susceptible to conventional antibiotics and mainly susceptible to lantibiotic bacteriocins, namely, gallidermin and nisin; they even showed a higher susceptibility to gallidermin than to nisin. They were also susceptible to enterocin–durancin, but most strains showed resistance to enterocin A/P. This study indicated that bacteriocins can play a key role in preventing and/or protecting from undesirable bacterial multiplication or contamination in the food industry and that they have great potential for further experimental applications.


Author(s):  
Kamni Rajput ◽  
Ramesh Chandra Dubey

In this paper, an investigation on lactic acid bacterial isolates from ethnic goat raw milk samples were examined for their probiotic potential and safety parameters. For this purpose, isolated bacterial cultures were screened based on certain parameters viz., sugar fermentation, tolerance to temperature, salt, low pH, bile salts, and phenol resistance. After that, these bacterial cultures were more estimated in vitro for auto-aggregation, cell surface hydrophobicity, response to simulated stomach duodenum channel, antibiotic resistance, and antimicrobial activity. Besides, probiotic traits show the absence of gelatinase and hemolytic activity supports its safety. The isolate G24 showed good viability at different pH, bile concentration, phenol resistance and response to simulated stomach duodenum passage but it did not show gelatinase and hemolytic activities. Isolate G24 was susceptible to amikacin, carbenicillin, kanamycin, ciprofloxacin, co-trimazine, nitrofurantoin, streptomycin, and tetracycline. Isolate G24 also exhibited antimicrobial action against five common pathogenic bacteria, such as Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Listeria monocytogens, and Salmonella typhimurium. It displayed the maximum auto-aggregation, cell surface hydrophobicity to different hydrocarbons. Following molecular characterization the isolate G24 was identified as Enterococcus hirae with 16S rRNA gene sequencing and phylogeny. E. hirae G24 bears the excellent properties of probiotics.


PLoS Biology ◽  
2020 ◽  
Vol 18 (12) ◽  
pp. e3000987
Author(s):  
Clare L. Kinnear ◽  
Elsa Hansen ◽  
Valerie J. Morley ◽  
Kevin C. Tracy ◽  
Meghan Forstchen ◽  
...  

The antimicrobial resistance crisis has persisted despite broad attempts at intervention. It has been proposed that an important driver of resistance is selection imposed on bacterial populations that are not the intended target of antimicrobial therapy. But to date, there has been limited quantitative measure of the mean and variance of resistance following antibiotic exposure. Here we focus on the important nosocomial pathogen Enterococcus faecium in a hospital system where resistance to daptomycin is evolving despite standard interventions. We hypothesized that the intravenous use of daptomycin generates off-target selection for resistance in transmissible gastrointestinal (carriage) populations of E. faecium. We performed a cohort study in which the daptomycin resistance of E. faecium isolated from rectal swabs from daptomycin-exposed patients was compared to a control group of patients exposed to linezolid, a drug with similar indications. In the daptomycin-exposed group, daptomycin resistance of E. faecium from the off-target population was on average 50% higher than resistance in the control group (n = 428 clones from 22 patients). There was also greater phenotypic diversity in daptomycin resistance within daptomycin-exposed patients. In patients where multiple samples over time were available, a wide variability in temporal dynamics were observed, from long-term maintenance of resistance to rapid return to sensitivity after daptomycin treatment stopped. Sequencing of isolates from a subset of patients supports the argument that selection occurs within patients. Our results demonstrate that off-target gastrointestinal populations rapidly respond to intravenous antibiotic exposure. Focusing on the off-target evolutionary dynamics may offer novel avenues to slow the spread of antibiotic resistance.


2020 ◽  
Vol 9 (4-s) ◽  
pp. 1218-1221 ◽  
Author(s):  
Dhaval M Patel ◽  
Advaita B Patel ◽  
Riddhi D Trivedi ◽  
Viram J Parmar ◽  
Urvisha V Bangoriya

Evaluation of the effect of hydroalcoholic extracts of Cassia occidentalis leaves in neutrophil adhesion test in Rats. The effect of oral administration of hydroalcoholic extract of Cassia occidentalis leaves on neutrophil has been studied and is compared with control group on rats. The differential leukocyte count (DLC) was performed by fixing the blood smears and staining with leucofine and percent neutrophils in each sample was determined. After the initial counts, blood samples were incubated with 80 mg/ml of nylon fibers for 10 min at 37ºC. Supplementation with hydroalcoholic extract of Cassia occidentalis leaves significantly decreased neutrophil adhesion. The results indicate that the leaf of Cassia occidentalis is endowed with protected neutrophil adhesion. These effects could conclude that Cassia occidentalis has an antiasthmatic property. Keywords: Cassia occidentalis, Neutrophil adhesion, analysis of variance


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