Synergistic Effects of Probiotics and Phytobiotics on the Intestinal Microbiota in Young Broiler Chicken

Probiotics and phytobiotics have been studied as in-feed antibiotic alternatives for decades, yet there are no studies on their possible symbiotic effects. In the present study, newly hatched chickens were fed with feeds supplemented either with host-specific Lactobacillus strains (L. agilis and L. salivarius), commercial phytobiotics, or combinations of both. After 13 days of life, crops and caecums were analyzed for bacterial composition (16S rDNA sequencing, qPCR) and activity (bacterial metabolites). Crop and caecum samples were also used to study the ex vivo survival of a broiler-derived extended-spectrum beta-lactamase (ESBL) producing Escherichia coli strain. In the crop, combinations of probiotics and phytobiotics, but not their single application, increased the dominance of lactobacilli. The single application of phytobiotics reduced the metabolite concentrations in the crop, but certain combinations synergistically upregulated the metabolites. Changes in the qualitative and quantitative composition of the caecal microbiota were less pronounced than in the crop. Acetate concentrations were significantly lower for phytobiotics or the L. agilis probiotic strain compared to the control group, but the L. salivarius probiotic showed significantly higher acetate concentrations alone or in combination with one phytobiotic. The synergistic effects on the reduction of the ex vivo survival of an ESBL producing E. coli strain in crop or caecum contents were also observed for most combinations. This study shows the beneficial synergistic effects of probiotics and phytobiotics on the intestinal bacterial composition and their metabolic activity in young broilers. The reduced survival of potentially problematic bacteria, such as ESBL-producing E. coli further indicates that combinations of probiotics and phytobiotics may lead to a more enhanced functionality than their individual supplementation.

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ENDOTOXINEMIA IN PATIENTS WITH OBESITY AND NON-ALCOHOLIC FATTY LIVER DISEASE

PROBLEMS OF ENDOCRINE PATHOLOGY ◽

10.21856/j-pep.2021.3.11 ◽

2021 ◽

Vol 77 (3) ◽

pp. 76-82

Author(s):

Galyna Fadieienko ◽

Oleksiy Gridnyev ◽

Inna Kushnir

Keyword(s):

Bacteroides Fragilis ◽

Control Group ◽

Quantitative Composition ◽

Gram Negative Bacteria ◽

Alcoholic Fatty Liver ◽

E Coli ◽

Bacteroides Fragilis Group ◽

Polymerase Chain ◽

Lactobacillus Spp ◽

Alcoholic Fatty Liver Disease

Objective — to study the features of endotoxinemia and its relationship with changes in the intestinal microbial spectrum in obese patients with non-alcoholic fatty liver disease (NAFLD). Materials and methods. 84 patients with obesity and NAFLD were examined. The control group consisted of 20 apparently healthy individuals. The concentration of endotoxin (ET) in the blood serum was determined using the LAL Chromogenic Endpoint Assay kit manufactured by Hycult Biotech (Netherlands), relative quantitative composition of the microbiota at the level of basic phylotypes — by real-time polymerase chain reaction (CFX96Touch (Bio-Rad, USA)) using universal primers for the 16SpPHK gene and taxon-specific primers), quantitative composition of microbiota — using test of the «Colonoflor-16» system (Alfalab, RF) by real-time polymerase chain reaction.Results. Among patients with comorbidity of obesity and NAFLD, there were 40.48 % of males and 59.52 % of females, whose mean age was 53.72 ± 4.61 years. The ET level in the examined patients with comorbidity of obesity and NAFLC was significantly (p < 0.001) higher (1.01 ET/ml) than in the control group (0.60 ET/ml), was significantly (p < 0,05) is higher in women than in men (1.06 and 0.92 UE/ml, respectively) and increased with increasing age of patients (r = 0.30, p < 0.05). The ET level correlated with the relative content of Firmicutes (r = 0.39, p < 0.05) and their ratio in Bacteroidetes (r = 0.29, p < 0.05) and the level of Bifidobacterium spp. (r = 0 .37, p < 0 .05) a nd h ad a n i nverse r elationship w ith t he r elative c ontent o f B acteroidetes ( r = – 0.42, p < 0.01), including Bacteroides fragilis group (r = – 0.43, p < 0.01), Escherichia coli (r = – 0.41, p < 0.01) and total bacterial mass (r = – 0.39, p < 0.05). In the intestinal microbiota of the examined patients, a decrease in the representatives of the Lactobacillus spp. and Bifidobacterium spp. in 84.5 % and 30.9 % of patients, respectively, as well as Bacteroides thetaiotaomicron (88.0 %), Akkermansia muciniphila (79.8 %) and Faecalibacteriumprausnitzii (33.3 %). Whereas the number of gram-negative bacteria increased primarily due to Enterobacter spp, Citrobacter spp (45.2 %), Escherichia coli (19 %), Bacteroides fragilis group (29.8 %). It should be noted that the decrease in the level of A. muciniphila was often accompanied by an increase in the content of enterobacteria, while the excess content of B. fragilis group was accompanied by an increase above the upper limit of E. coli and the presence of anaerobic imbalance in the microbiota. In turn, increasing the number of Enterobacter spp. / Citrobacter spp. It was often accompanied by an increase in the content of E. coli and representatives of the B. fragilis group, against the background of a reduced number of A. muciniphila. Conclusions. The above indicates the presence of metabolic endotoxinemia in patients with comorbidity of obesity and NAFLD against the background of a decrease in the number of gram-positive anaerobic Lactobacillus spp. i Bifidobacterium spp. and bacteria involved in providing the intestinal barrier function (Bacteroides Thetaiotaomicron, Akkermansia muciniphila, Faecalibacterium prausnitzii), which creates conditions for an increased amount of ET into the blood and is especially important against the background of an increase in the content of gram-negative bacteria (representatives of gammaproteobacteria and bacteria of the Bacteroides fragilis), which are the source of ET.

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Novel ex vivo screening assay to preselect farm specific pre- and probiotics in pigs

Beneficial Microbes ◽

10.3920/bm2020.0226 ◽

2021 ◽

pp. 1-16

Author(s):

K. Zeilinger ◽

J. Hellmich ◽

J. Zentek ◽

W. Vahjen

Keyword(s):

Ex Vivo ◽

Screening Method ◽

Coli Strain ◽

Screening Assay ◽

Future Studies ◽

E Coli ◽

Feed Management ◽

Specific Pathogen ◽

On Farm ◽

Ex Vivo Assay

A novel rapid ex vivo assay was developed as part of a concept to determine potential tailor-made combinations of pre- and probiotics for individual farms. Sow faecal slurries from 20 German pig farms were anaerobically incubated with pre- and probiotics or their combinations together with pathogenic strains that are of interest in pig production. Aliquots of these slurries were then incubated with media containing antibiotic mixtures allowing only growth of the specific pathogen. Growth was monitored and lag time was used to determine the residual fitness of the pathogenic strains. The background growth could be inhibited for an Escherichia coli- and a Clostridium difficile- but not for a Clostridium perfringens strain. The prebiotic fructo-oligosaccharides (FOS) and its combination with probiotics reduced the residual fitness of the E. coli strain in some farms. However, notable exceptions occurred in other farms where FOS increased the fitness of the E. coli strain. Generally, combinations of pre- and probiotics did not show additive effects on fitness for E. coli but displayed farm dependent differences. The effects of pre- and probiotics on the residual fitness of the C. difficile strain were less pronounced, but distinct differences between single application of prebiotics and their combination with probiotics were observed. It was concluded that the initial composition of the microbiota in the samples was more determinative for incubations with the C. difficile strain than for incubations with the E. coli strain, as the presumed fermentation of prebiotic products showed less influence on the fitness of the C. difficile strain. Farm dependent differences were pronounced for both pathogenic strains and therefore, this novel screening method offers a promising approach for pre-selecting pre- and probiotics for individual farms. However, evaluation of farm metadata (husbandry, feed, management) will be crucial in future studies to determine a tailor-made solution for combinations of pre- and probiotics for individual farms. Also, refinement of the ex vivo assay in terms of on-farm processing of samples and validation of unambiguous growth for pathogenic strains from individual farms should be addressed.

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Human Neutrophil Chemotaxis Is Modulated by Capsule and O Antigen from an Extraintestinal Pathogenic Escherichia coli Strain

Infection and Immunity ◽

10.1128/iai.71.11.6435-6445.2003 ◽

2003 ◽

Vol 71 (11) ◽

pp. 6435-6445 ◽

Cited By ~ 9

Author(s):

Thomas A. Russo ◽

Bruce A. Davidson ◽

Diana M. Topolnycky ◽

Ruth Olson ◽

Stacy A. Morrill ◽

...

Keyword(s):

Escherichia Coli ◽

Ex Vivo ◽

Specific Antigen ◽

Lavage Fluid ◽

Coli Strain ◽

Neutrophil Chemotaxis ◽

Gram Negative ◽

E Coli ◽

O Antigen

ABSTRACT Gram-negative enteric bacilli, such as Escherichia coli, are common causes of nosocomial pneumonia. The interaction between pulmonary neutrophils and the infecting pathogen is a critical step in determining the outcome. Previous studies from our laboratory, for which a rat model of pneumonia was used, established that pulmonary neutrophil recruitment was modulated by the E. coli virulence factors capsule and O-specific antigen. To begin to understand the mechanism by which this recruitment occurs, we conducted in vitro and ex vivo chemotaxis assays, for which we used a clinically relevant E. coli isolate (CP9) and isogenic derivatives that were deficient in only the O antigen (CP921) or capsule (CP9.137) as chemoattractants with or without the high-affinity N-formylmethionyl-leucyl-phenylalanine receptor antagonist N-tert-butoxycarbonyl-methionine-leucine-phenylalanine (N-t-BOC). Given that only live E. coli was used for the initial in vitro chemotaxis assays, it was predicted that only N-t-BOC-sensitive chemotaxis would occur. However, both N-t-BOC-sensitive and -insensitive chemotaxis was observed. N-t-BOC-insensitive chemotaxis was mediated in part by interleukin 8, which was produced by neutrophils that had migrated toward E. coli. N-t-BOC-insensitive chemotaxis was only observed when live E. coli bacteria, not cell-free E. coli culture supernatants, were used as chemoattractants, suggesting that a direct E. coli-neutrophil interaction was necessary. The presence of both capsule and O antigen diminished total, N-t-BOC-sensitive, and N-t-BOC-insensitive neutrophil chemotaxis in vitro. The presence of capsule significantly decreased total, N-t-BOC-sensitive, and N-t-BOC-insensitive neutrophil chemotaxis ex vivo when cell-free bronchoalveolar lavage fluid from infected rats was used as the source of chemotactic factors. These effects of E. coli capsule and O antigen on neutrophil chemotaxis are novel, and they expand our understanding of the mechanisms by which these virulence traits contribute to the pathogenesis of gram-negative pneumonia and other extraintestinal infections.

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Porcine Protein Hydrolysates (PEPTEIVA®) Promote Growth and Enhance Systemic Immunity in Gilthead Sea Bream (Sparus aurata)

Animals ◽

10.3390/ani11072122 ◽

2021 ◽

Vol 11 (7) ◽

pp. 2122

Author(s):

Enric Gisbert ◽

Antoni Ibarz ◽

Joana P. Firmino ◽

Laura Fernández-Alacid ◽

Ricardo Salomón ◽

...

Keyword(s):

Feed Efficiency ◽

Sparus Aurata ◽

Ex Vivo ◽

Protein Hydrolysate ◽

Sea Bream ◽

Control Group ◽

Skin Mucus ◽

E Coli ◽

Tnf Α ◽

Cellular Immune

The effects of porcine plasma protein hydrolysate (PPH) on growth, feed efficiency, and immune responses was evaluated in Sparus aurata. Fish were fed two isoproteic (48% protein), isolipidic (17% fat), and isoenergetic diets (21.7 MJ/kg) diets, one of them containing 5% PPH at the expense of fishmeal. Both diets were tested for 92 days. A significant increase in growth was observed in fish fed the PPH diet in comparison to the control group (182.2 ± 4.4 vs. 173.8 ± 4.1 g), as well as an increase in feed intake without worsening FCR values. An ex vivo assay, with splenocytes incubated with lipopolysaccharide, was conducted to evaluate the cellular immune competence of fish. Genes involved in humoral immunity (lys, IgM), pro- (tnf-α, il-1β), and anti-inflammatory (tgf-β1, il10) cytokines were upregulated in the PPH group in comparison to the control group. The inclusion of PPH in diets enhanced the antibacterial capacity of skin mucus, as the co-culture of selected bacteria (E. coli, V. anguillarum, and P. anguilliseptica) with skin mucus indicated. The present results showed that the PPH in low fishmeal diets (2%) promoted growth and feed efficiency, as well as enhancing the immune response, which indicates that this is a safe and functional ingredient for aquafeeds.

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Identity of blaCTX-M Carrying Plasmids in Sequential ESBL-E. coli Isolates from Patients with Recurrent Urinary Tract Infections

Microorganisms ◽

10.3390/microorganisms9061138 ◽

2021 ◽

Vol 9 (6) ◽

pp. 1138

Author(s):

Nahid Karami ◽

Sriram KK ◽

Shora Yazdanshenas ◽

Yii-Lih Lin ◽

Daniel Jaén-Luchoro ◽

...

Keyword(s):

Urinary Tract ◽

Urinary Tract Infections ◽

Rare Event ◽

Coli Strain ◽

Health It ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Recurrent Urinary Tract Infections ◽

Global Threat ◽

Tract Infections

Plasmid-mediated multidrug resistance in E. coli is becoming increasingly prevalent. Considering this global threat to human health, it is important to understand how plasmid-mediated resistance spreads. From a cohort of 123 patients with recurrent urinary tract infections (RUTI) due to extended spectrum beta-lactamase (ESBL)-producing Escherichia coli (ESBL E. coli), only five events with a change of ESBL E. coli strain between RUTI episodes were identified. Their blaCTX-M encoding plasmids were compared within each pair of isolates using optical DNA mapping (ODM) and PCR-based replicon typing. Despite similar blaCTX-M genes and replicon types, ODM detected only one case with identical plasmids in the sequential ESBL E. coli strains, indicating that plasmid transfer could have occurred. For comparison, plasmids from seven patients with the same ESBL E. coli strain reoccurring in both episodes were analyzed. These plasmids (encoding blaCTX-M-3, blaCTX-M-14, and blaCTX-M-15) were unaltered for up to six months between recurrent infections. Thus, transmission of blaCTX-M plasmids appears to be a rare event during the course of RUTI. Despite the limited number (n = 23) of plasmids investigated, similar blaCTX-M-15 plasmids in unrelated isolates from different patients were detected, suggesting that some successful plasmids could be associated with specific strains, or are more easily transmitted.

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Antimicrobial and Cytotoxic activity of rhizome extract of Acorus calamus (Bojho) in combination with different antimicrobial agents: Synergistic Effects

International Journal of Applied Sciences and Biotechnology ◽

10.3126/ijasbt.v7i3.25717 ◽

2019 ◽

Vol 7 (3) ◽

pp. 347-353

Author(s):

Hemanta Khanal

Keyword(s):

Plant Extract ◽

Membrane Filtration ◽

Antimicrobial Agents ◽

Synergistic Effects ◽

Acorus Calamus ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Soxhlet Apparatus ◽

Sweet Flag ◽

Ethanolic Extraction

Acorus calamus (sweet flag) is a monocot plant found in wetland, have the scented leaves and rhizomes. Various active bio-ingredients of A. calamus had been studied and characterized and some of them are known for antimicrobial and antitumor activities. The dry rhizomes were powdered ethanolic extraction was performed in a Soxhlet apparatus. The extract was dried and re-suspended to sterile distilled water and sterilized by membrane filtration. The synergist affects against bacteria, fungi, helminths were evaluated and cytotoxic assay was performed. The minimum inhibitory concentrations of plant extract was found to be 50µg/ml for the vancomycin resistance Staphylococcus aureus, for extended spectrum beta lactamase Escherichia coli, Methicillin resistant S. aureus, were 100 µg/ml. Synergistically cefixime and cefpodoxime both antibiotics are found to be effective against all strains of S. aureus and E. coli except VRSA. The antifungal characteristics were found to be effective when agar cup diffusion were performed in combination with fluconazole antifungal drug. The combination of plant extract was more effective anthelminthic drug than the anthelminthic drug alone. Similarly, the plant extract has lethal concentration 50 (LC50) was found to be 173.3 µg / ml and LC90 was 555. 4 µg / ml on brine shrimp. The A. calamus has potential characteristics to be as antimicrobial and antitumor medicine when used synergistically with antimicrobial agents.

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