![Page 1: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/1.jpg)
Rita Rahmeh
ANTIMICROBIAL ACTIVITY OF
LACTIC ACID BACTERIA AND
ANTIMICROBIAL PEPTIDES FROM
CAMEL MILK
1
![Page 2: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/2.jpg)
The Microbiome Confers Health Benefits
Probiotics
“Live microorganisms that, when administered in
adequate amounts, confer a health benefit on the host”
(International Scientific Association for Probiotics and
Prebiotics).
Molecules that have broad activity to directly kill
bacteria, yeasts, fungi, and viruses.
Bacteriocins: produced by lactic acid bacteria
Antimicrobial
peptides
2
The microbiome of bovine and non-bovine
milk is dominated by lactic acid bacteria
![Page 3: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/3.jpg)
Reducing:
•Antibiotic resistant bacteria
•Economic losses
•Negative impact on human health
Commercialization
Microbiome Research Impacts Commercialization
3•2015, probiotic market $33.19 billion
•2020, expected to reach $46.55 billion
Probiotics
Research
Animal Production Food Preservation
Bacteriocins
![Page 4: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/4.jpg)
Probiotics
Bacteriocins
Chicken or Turkey
feed
Campylobacter
Salmonella
Probiotics Improve Feed and Food
Feed Improvement
Food Improvement
Nisin A Nisaplin
Bacteriocin
Commercial food
preservative
50 countries (U.S. and
EU)
4
![Page 5: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/5.jpg)
Camel Milk as a Natural Source of Probiotics
5
•Nutritional value
•Health benefits (boosts the immune system, aids in
preventing diabetes and anemia, helps in reducing allergy
such as lactose intolerance, etc.)
•Worthy of attention internationally
•Source of novel antimicrobial peptides
![Page 6: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/6.jpg)
Objective: Isolate Probiotics from Camel Milk
Lactic acid bacteria
Food technology
Food preservatives
Bacteriocins
Farms
Feed additives
6
![Page 7: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/7.jpg)
7
Lactic acid bacteria isolation
Isolation and Characterization of Lactic Acid Bacteria
Gram Staining
Characterization of lactic acid bacteria
Gram-Positive Gram-Negative
Catalase
2 H2O2 2 H2O + O2
Catalase Test
![Page 8: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/8.jpg)
Genus Species
Enterococcus
Enterococcus faecium
Enterococcus gallinarum
Enterococcus durans
Lactococcus
Lactococcus lactis
Lactococcus garvieae
Lactococcus brevis
Pediococcus
Pediococcus pentosaceus
Pediococcus acidilactici
Lactobacillus
Lactobacillus reuteri
Lactobacillus fermentum
Lactobacillus salivarius
Lactobacillus plantarum
Weissella
Weissella confusa
Weissella sp. t4r2c13
Leuconostoc
Leuconostoc pseudomesenteroides
Streptococcus
Streptococcus infantarius
Seven Genus of Lactic Acid Bacteria Were Identified
8
Rahmeh et al., Probiotics and Antimicrobial Proteins (submitted)
![Page 9: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/9.jpg)
9
Inoculate the agar plate with a spot of lactic acid bacteria
Incubate at the growth conditions of lactic acid bacteria
Add second layer of agar inoculated with pathogen
Incubate at the growth conditions of pathogen
Lactic Acid Bacteria Exhibit Inhibitory Activity
Spot-on-the-lawn method
9
![Page 10: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/10.jpg)
Salmonella enterica ATCC 13076
Escherichia coli ATCC 25922
Staphylococcus aureus ATCC 25923
Staphylococcus epidermidis ATCC 12228
Listeria monocytogenes ATCC 7644
Shigella ATCC 12022
Pseudomonas aeruginosa CUGG 060519
Multi Drug Resistant Salmonella10
Pathogens causing human, animal diseases and food spoilage
28 Lactic Acid Bacteria Exhibit Inhibitory Activity
![Page 11: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/11.jpg)
11
Inoculate the agar plate with pathogen
Fill the wells with the cell-free supernatant of lactic acid bacteria
Incubate at the growth conditions of pathogen
Four Lactic Acid Bacteria Produce Bacteriocins
Agar diffusion method
Listeria monocytogenes ATCC
7644
11
![Page 12: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/12.jpg)
0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24
0
100
200
300
400
500
600
Volume (mL)
Ab
so
rban
ce a
t 280 (
mA
U)
Gel filtration using Superdex 75 10/300 GL
17.9 ml-18.9 ml
12
Purification of Bacteriocins
Rahmeh et al., J Dairy Sci. 2018 Jun;101(6):4944-4952
![Page 13: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/13.jpg)
Treatment Antimicrobial Activity
of Bacteriocin
Enzymes
Control ++
α-chymotrypsin -
Proteinase K -
Papain -
Trypsin -
Protease -
pH
Control ++
2 ++
6 ++
8 ++
10 ++
Heat/Time
Control ++
37°C/180 min ++
60°C/60 min ++
80°C/60 min ++
100°C/30 min +
121°C/15 min +
13
Characterization of Bacteriocins
![Page 14: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/14.jpg)
14
Microbiome of camel milk is a promising source of probiotics
Food preservatives
Feed Additives
Improvement in Food and Feed Production
![Page 15: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/15.jpg)
Application Challenges in The Food System
Probiotics:
•The growth of lactic acid bacteria or the production of
bacteriocins are affected by pH, temperature fluctuation
during food processing, etc.
•Other bacterial fermenters co-inoculated negatively impact
bacteriocins production and biopreservation
Bacteriocins
The efficacy of pure bacteriocins is affected by:
• pH of the food matrix which influences its solubility
•Enzymes that inactivate the bacteriocins
•Food additives (spices and salt) which greatly reduce its
efficacy
15
![Page 16: ANTIMICROBIAL ACTIVITY OF L ACID BACTERIA AND](https://reader035.vdocuments.us/reader035/viewer/2022081416/629e23c68e631f06b5533199/html5/thumbnails/16.jpg)
Thank you
16