scholarly journals Multivalent Chromosomal Expression of the Clostridium botulinum Serotype A Neurotoxin Heavy-Chain Antigen and the Bacillus anthracis Protective Antigen in Lactobacillus acidophilus

2016 ◽  
Vol 82 (20) ◽  
pp. 6091-6101 ◽  
Author(s):  
Sarah O'Flaherty ◽  
Todd R. Klaenhammer
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Bacillus Anthracis ◽  
Lactobacillus Acidophilus ◽  
Clostridium Botulinum ◽  
Protective Antigen ◽  
Vaccine Delivery ◽  
Mucosal Vaccine ◽  
Expression Systems ◽  
Content Type

ABSTRACTClostridium botulinumandBacillus anthracisproduce potent toxins that cause severe disease in humans. New and improved vaccines are needed for both of these pathogens. For mucosal vaccine delivery using lactic acid bacteria, chromosomal expression of antigens is preferred over plasmid-based expression systems, as chromosomal expression circumvents plasmid instability and the need for antibiotic pressure. In this study, we constructed three strains ofLactobacillus acidophilusNCFM expressing from the chromosome (i) the nontoxic host receptor-binding domain of the heavy chain ofClostridium botulinumserotype A neurotoxin (BoNT/A-Hc), (ii) the anthrax protective antigen (PA), and (iii) both the BoNT/A-Hc and the PA. The BoNT/A-Hc vaccine cassette was engineered to contain the signal peptide from the S-layer protein A fromL. acidophilusand a dendritic-cell-targeting peptide. A chromosomal region downstream oflba0889carrying a highly expressed enolase gene was selected for insertion of the vaccine cassettes. Western blot analysis confirmed the heterologous expression of the two antigens from plasmid and chromosome locations. Stability assays demonstrated loss of the vaccine cassettes from expression plasmids without antibiotic maintenance. RNA sequencing showed high expression of each antigen and that insertion of the vaccine cassettes had little to no effect on the transcription of other genes in the chromosome. This study demonstrated that chromosomal integrative recombinant strains are promising vaccine delivery vehicles when targeted into high-expression chromosomal regions. Levels of expression match high-copy-number plasmids and eliminate the requirement for antibiotic selective maintenance of recombinant plasmids.IMPORTANCEClostridium botulinumandBacillus anthracisproduce potent neurotoxins that pose a biochemical warfare concern; therefore, effective vaccines against these bacteria are required. Chromosomal expression of antigens is preferred over plasmid-based expression systems since expressing antigens from a chromosomal location confers an advantage to the vaccine strains by eliminating the antibiotic maintenance required for plasmids and negates issues with plasmid instability that would result in loss of the antigen. Lactic acid bacteria, includingLactobacillus acidophilus, have shown potential for mucosal vaccine delivery, asL. acidophilusis bile and acid tolerant, allowing transit through the gastrointestinal tract where cells interact with host epithelial and immune cells, including dendritic cells. In this study, we successfully expressedC. botulinumandB. anthracisantigens in the probioticL. acidophilusstrain NCFM. Both antigens were highly expressed individually or in tandem from the chromosome ofL. acidophilus.

scholarly journals Lactic Acid Bacteria as Vectors: A Novel Approach for Mucosal Vaccine Delivery

2018 ◽  
Vol 09 (02) ◽  
Author(s):  
Beenish Israr ◽  
Jaehan Kim ◽  
Sidra Anam ◽  
Faisal Rasheed Anjum
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Vaccine Delivery ◽  
Mucosal Vaccine ◽  
Novel Approach

Bacteriocin Inhibition of Clostridium botulinum Spores by Lactic Acid Bacteria

1991 ◽  
Vol 54 (5) ◽  
pp. 349-353 ◽  
Author(s):  
AMECHI OKEREKE ◽  
THOMAS J. MONTVILLE
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactobacillus Acidophilus ◽  
Clostridium Botulinum ◽  
Lytic Phage ◽  
Pediococcus Pentosaceus ◽  
Inhibitory Cell ◽  
Simultaneous Testing ◽  
Inhibition Zones

Twenty-three strains of lactic acid bacteria were tested by deferred antagonism methods for bacteriocin-like activity against types A and B spores from 11 proteolytic and nonproteolytic Clostridium botulinum strains. Pediococcus pentosaceus ATCC 43200, Pediococcus pentosaceus ATCC 43201, Lactococcus lactis subsp. lactis ATCC 11454, Lactobacillus acidophilus N2, Lactobacillus plantarum Lb75, Lactobacillus plantarum Lb592, and Lactobacillus plantarum BN exhibited bacteriocin-like inhibition of all C. botulinum strains tested. By excluding inhibition due to hydrogen peroxide, acid, and lytic phage and confirming their proteinaceous nature, the inhibitors were confirmed as bacteriocins. The minimum inhibitory cell concentrations (MICC) required to produce 1 mm radius inhibition zones were determined by direct antagonism testing. Only strains 43200, 43201, 11454, and N2 were inhibitory when cultured simultaneously with the botulinal spores. The MICCs of strains antagonistic to C. botulinum spores by simultaneous testing ranged between 1.6 × 105and 4.7 × 107CFU/ml. Based on the MICCs, P. pentosaceus 43200 was most inhibitory to C. botulinum.

Inhibition of Nonproteolytic Clostridium botulinum with Lactic Acid Bacteria and Their Bacteriocins at Refrigeration Temperatures

2003 ◽  
Vol 66 (4) ◽  
pp. 674-678 ◽  
Author(s):  
S. RODGERS ◽  
P. PEIRIS ◽  
G. CASADEI
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Prussian Blue ◽  
Lactobacillus Acidophilus ◽  
Resistant Strain ◽  
Clostridium Botulinum ◽  
Incubation Temperature ◽  
Inhibition Zone ◽  
Zone Size

Nonproteolytic Clostridium botulinum (strains 17B, Beluga, and 202F) was found to be inhibited by Lactobacillus, Lactococcus, Streptococcus, and Pediococcus species in tests by the spot-on-the-lawn simultaneous-antagonism method at 10, 15, and 25°C. C. botulinum 17B was the most resistant strain. Inhibition zone size increased with decreasing incubation temperature. Six strains of Lactobacillus acidophilus and seven strains of bifidobacteria failed to produce an inhibition zone on buffered reinforced clostridium Prussian blue agar seeded with spores of any of the selected C. botulinum strains. C. botulinum 17B was sensitive to 50 to 100 IU of nisin per ml and to 10 to 20 AU of pediocin A per ml.

scholarly journals Core Fluxome and Metafluxome of Lactic Acid Bacteria under Simulated Cocoa Pulp Fermentation Conditions

2013 ◽  
Vol 79 (18) ◽  
pp. 5670-5681 ◽  
Author(s):  
Philipp Adler ◽  
Christoph Josef Bolten ◽  
Katrin Dohnt ◽  
Carl Erik Hansen ◽  
Christoph Wittmann
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Starter Cultures ◽  
Superior Performance ◽  
Gas Chromatography Mass Spectrometry ◽  
Glycolytic Flux ◽  
Content Type ◽  
Phosphoketolase Pathway ◽  
Labeling Approach ◽  
Cocoa Fermentation

ABSTRACTIn the present work, simulated cocoa fermentation was investigated at the level of metabolic pathway fluxes (fluxome) of lactic acid bacteria (LAB), which are typically found in the microbial consortium known to convert nutrients from the cocoa pulp into organic acids. A comprehensive13C labeling approach allowed to quantify carbon fluxes during simulated cocoa fermentation by (i) parallel13C studies with [13C6]glucose, [1,2-13C2]glucose, and [13C6]fructose, respectively, (ii) gas chromatography-mass spectrometry (GC/MS) analysis of secreted acetate and lactate, (iii) stoichiometric profiling, and (iv) isotopomer modeling for flux calculation. The study of several strains ofL. fermentumandL. plantarumrevealed major differences in their fluxes. TheL. fermentumstrains channeled only a small amount (4 to 6%) of fructose into central metabolism, i.e., the phosphoketolase pathway, whereas onlyL. fermentumNCC 575 used fructose to form mannitol. In contrast,L. plantarumstrains exhibited a high glycolytic flux. All strains differed in acetate flux, which originated from fractions of citrate (25 to 80%) and corresponding amounts of glucose and fructose. Subsequent, metafluxome studies with consortia of differentL. fermentumandL. plantarumstrains indicated a dominant (96%) contribution ofL. fermentumNCC 575 to the overall flux in the microbial community, a scenario that was not observed for the other strains. This highlights the idea that individual LAB strains vary in their metabolic contribution to the overall fermentation process and opens up new routes toward streamlined starter cultures.L. fermentumNCC 575 might be one candidate due to its superior performance in flux activity.

Cell-free supernatants produced by lactic acid bacteria reduce Salmonella population in vitro

Microbiology ◽  
2021 ◽  
Vol 167 (11) ◽  
Author(s):  
Alberto Gonçalves Evangelista ◽  
Jessica Audrey Feijó Corrêa ◽  
João Vitor Garcia dos Santos ◽  
Eduardo Henrique Custódio Matté ◽  
Mônica Moura Milek ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Lactic Acid ◽  
Antimicrobial Resistance ◽  
Lactic Acid Bacteria ◽  
Type Species ◽  
Feed Additives ◽  
Poultry Feed ◽  
Content Type ◽  
Link Type

The genus Salmonella is closely associated with foodborne outbreaks and animal diseases, and reports of antimicrobial resistance in Salmonella species are frequent. Several alternatives have been developed to control this pathogen, such as cell-free supernatants (CFS). Our objective here was to evaluate the use of lactic acid bacteria (LAB) CFS against Salmonella in vitro. Seventeen strains of LAB were used to produce CFS, and their antimicrobial activity was screened towards six strains of Salmonella . In addition, CFS were also pH-neutralized and/or boiled. Those with the best results were lyophilized. MICs of lyophilized CFS were 11.25–22.5 g l–1. Freeze-dried CFS were also used to supplement swine and poultry feed (11.25 g kg–1) and in vitro simulated digestion of both species was performed, with Salmonella contamination of 5×106 and 2×105 c.f.u. g−1 of swine and poultry feed, respectively. In the antimicrobial screening, all acidic CFS were able to inhibit the growth of Salmonella . After pH neutralization, Lactobacillus acidophilus Llorente, Limosilactobacillus fermentum CCT 1629, Lactiplantibacillus plantarum PUCPR44, Limosilactobacillus reuteri BioGaia, Lacticaseibacillus rhamnosus ATCC 7469 and Pediococcus pentosaceus UM116 CFS were the only strains that partially maintained their antimicrobial activity and, therefore, were chosen for lyophilization. In the simulated swine digestion, Salmonella counts were reduced ≥1.78 log c.f.u. g–1 in the digesta containing either of the CFS. In the chicken simulation, a significant reduction was obtained with all CFS used (average reduction of 0.59±0.01 log c.f.u. ml–1). In general, the lyophilized CFS of L. fermentum CCT 1629, L. rhamnosus ATCC 7469 and L. acidophilus Llorente presented better antimicrobial activity. In conclusion, CFS show potential as feed additives to control Salmonella in animal production and may be an alternative to the use of antibiotics, minimizing problems related to antimicrobial resistance.

Physicochemical, microbiological, and sensory characteristics of probiotic Bulgarian yoghurts obtained by ultrafiltration of goat’s milk

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mariya Dushkova ◽  
Siyka Kodinova ◽  
Zapryana Denkova ◽  
Velichka Yanakieva ◽  
Nikolay Dimitrov Menkov
Keyword(s):  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Acidophilus ◽  
Dry Matter ◽  
Titratable Acidity ◽  
Bifidobacterium Bifidum ◽  
Sensory Characteristics ◽  
Viable Cells ◽  
Goat’S Milk ◽  
Goat's Milk

Abstract The purpose of this study was to investigate the microbiological (number of viable lactic acid bacteria and bifidobacteria), physicochemical (dry matter, protein and fat contents, titratable acidity, and pH), and sensory characteristics (appearance of coagulum, taste and aroma, structure at cutting, color, and consistency at shattering) of probiotic Bulgarian yoghurts obtained by ultrafiltration of goat’s milk. These yoghurts were obtained using volume reduction ratios of 2 and 3 with the probiotic starters MZ2f, MZ2f + Bifidobacterium bifidum BB – 87, and MZ2f + Lactobacillus acidophilus LAB – 8. The increase in the level of the concentration by ultrafiltration led to an increase in the dry matter, protein and fat contents of the yoghurts, in the number of lactic acid bacteria and titratable acidity, and to a decrease in the pH. The twofold concentration by ultrafiltration resulted in a higher number of lactic acid bacteria in comparison with yoghurts made without ultrafiltration, and with better sensory characteristics compared to yoghurts without and with threefold ultrafiltration. The higher number of viable cells and better sensory characteristics were obtained for yoghurts with MZ2f + Bifidobacterium bifidum BB – 87 and MZ2f + Lactobacillus acidophilus LAB – 8 in comparison with MZ2f alone.

scholarly journals The Fluorocycline TP-271 Is Efficacious in Models of Aerosolized Bacillus anthracis Infection in BALB/c Mice and Cynomolgus Macaques

2017 ◽  
Vol 61 (10) ◽  
Author(s):  
Trudy H. Grossman ◽  
Michael S. Anderson ◽  
Lindsay Drabek ◽  
Melanie Gooldy ◽  
Henry S. Heine ◽  
...  
Keyword(s):  
Bacillus Anthracis ◽  
Protective Antigen ◽  
Day Treatment ◽  
Inhalation Exposure ◽  
Treated Group ◽  
Average Dose ◽  
Once Daily ◽  
Content Type ◽  
Cynomolgus Macaques ◽  
Observation Period

ABSTRACT The fluorocycline TP-271 was evaluated in mouse and nonhuman primate (NHP) models of inhalational anthrax. BALB/c mice were exposed by nose-only aerosol to Bacillus anthracis Ames spores at a level of 18 to 88 lethal doses sufficient to kill 50% of exposed individuals (LD50). When 21 days of once-daily dosing was initiated at 24 h postchallenge (the postexposure prophylaxis [PEP] study), the rates of survival for the groups treated with TP-271 at 3, 6, 12, and 18 mg/kg of body weight were 90%, 95%, 95%, and 84%, respectively. When 21 days of dosing was initiated at 48 h postchallenge (the treatment [Tx] study), the rates of survival for the groups treated with TP-271 at 6, 12, and 18 mg/kg TP-271 were 100%, 91%, and 81%, respectively. No deaths of TP-271-treated mice occurred during the 39-day posttreatment observation period. In the NHP model, cynomolgus macaques received an average dose of 197 LD50 of B. anthracis Ames spore equivalents using a head-only inhalation exposure chamber, and once-daily treatment of 1 mg/kg TP-271 lasting for 14 or 21 days was initiated within 3 h of detection of protective antigen (PA) in the blood. No (0/8) animals in the vehicle control-treated group survived, whereas all 8 infected macaques treated for 21 days and 4 of 6 macaques in the 14-day treatment group survived to the end of the study (56 days postchallenge). All survivors developed toxin-neutralizing and anti-PA IgG antibodies, indicating an immunologic response. On the basis of the results obtained with the mouse and NHP models, TP-271 shows promise as a countermeasure for the treatment of inhalational anthrax.

scholarly journals Complete Genome Sequence of Lactobacillus reuteri Byun-re-01, Isolated from Mouse Small Intestine

2018 ◽  
Vol 7 (17) ◽  
Author(s):  
Dongjun Kim ◽  
Mun-ju Cho ◽  
Seungchan Cho ◽  
Yongjun Lee ◽  
Sung June Byun ◽  
...  
Keyword(s):  
Small Intestine ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Genome Sequence ◽  
Complete Genome Sequence ◽  
Complete Genome ◽  
Lactobacillus Reuteri ◽  
Probiotic Bacteria ◽  
Content Type ◽  
Mouse Small Intestine

Lactic acid bacteria (LAB) are generally recognized as safe (GRAS) and serve as probiotic bacteria when consumed in adequate amounts. Here, we report the complete genome sequence of Lactobacillus reuteri Byun-re-01, isolated from mouse small intestine.

Sign in / Sign up

Export Citation Format

Share Document