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result(s) for
"Rizkinata, Denny"
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Synergistic action of bacteriophage and metabolites of Pseudomonas fluorescens JB3B and Streptomyces thermocarboxydus 18PM against Enterotoxigenic Escherichia coli and Bacillus cereus and their biofilm
by
Waturangi, Diana Elizabeth
,
Rizkinata, Denny
,
Yulandi, Adi
in
Acetic acid
,
Acids
,
Actinomycetes
2024
Background
Foodborne disease and food spoilage are the prime public health issue and food security round the globe. Significant disease outbreaks mostly linked to the existence of pathogenic bacteria that extremely challenging due to the persistence of biofilm-forming. Proteins and bacterial metabolites have been shown to have good antibacterial activity and effectively removal bacterial biofilm. Recently, bacteriophage and their encoded lytic proteins such as lysin have attracted attention as potential alternative agent to control undesirable pathogens in human body infection, increasing food safety as advance preservations and medical treatment such as phage therapy. For these reasons, the efficacy of bacteriophage and their potential in combination with bacterial metabolites from
Phyllosphere
and
Actinomycetes
bacteria (
Pseudomonas fluorescens
JB3B and
Streptomyces thermocarboxydus
18PM crude extracts) was the aim of this present study.
Results
In this study, bacteriophage BC-VP (1.28
±
0.29 × 10
11
PFU/ml) and ETEC-phage-TG (8.9
±
2.19 × 10
8
PFU/ml) isolated from artificial lake water from previous study showed potential activity to control
Bacillus cereus
(BC) and Enterotoxigenic
Escherichia coli
(ETEC) population. The combination of BC-VP with metabolite (
P. fluorescens
JB3B and
S. thermocarboxydus
18PM) which were known from previous study had antibiofilm activities were able to inhibit (86.1%; 83.3%) and destruct (41%; 45.5%) biofilm formation of
B. cereus
respectively. Likewise, the synergy of bacteriophage ETEC-phage-TG with the same crude extract also showed promising activity against biofilm of ETEC with percentage of inhibition (81.9%; 76.4%) and percentage of destruction (54.1%; 44.4%). Application in various food, combination of BC-VP and bacterial metabolite extract (
P. fluorescens
JB3B;
S. thermocarboxydus
18PM) were able to reduce
Bacillus cereus
population in mashed potato (99.6%; 99.4%) at cold temperature (4 °C) and (68.9%; 56.6%) at room temperature (28 °C), boiled pasta (99.5%; 99.4%) and (84.7%; 75.7%), also soymilk (96.9%; 96.7%) and (42.4%; 39.4%) respectively. Likewise, combination of ETEC-phage-TG and bacterial metabolite (
P. fluorescens
JB3B;
S. thermocarboxydus
18PM) potentially reduced ETEC population after two different temperatures (4 °C and 28 °C) incubation in bean sprouts (TFTC; TFTC) and (47.5%; 49.1%), chicken meat (TFTC; TFTC) and (58.1%; 54%), also minced beef (99.5%; 99.4%) and (41.1%; 28%). GC-MS determination performed, oxalic acid, phenol, phenylethyl alcohol, N-hexadecanoic acid, and pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl was the most active compound in
P. fluorescens
JB3B. 2,4-Di-tert-butylphenol, phenyl acetic acid, N-Hexadecanoic acid, pyrolol[1,2-a]pyrazine-1,4-dione, hexadro-3-92-methylpropyl, and Bis(2-ethylhexyl) phthalate was most active compound in the
S. thermocarboxydus
18PM isolates.
Conclusions
The combination of isolated bacteriophages and bacterial metabolite showed promising results to be used as biocontrol candidate to overcome biofilm formed by foodborne and food spoilage bacteria using their ability to produce antibiofilm compounds and lytic activity. In addition, this combination also potentially reduces the use or replace the drawbacks of common application such as antibiotic treatment.
Journal Article
Isolation and molecular characterization of bacteriophages isolated from lake water and their application in foods against Bacillus cereus
by
Rizkinata, Denny
,
Kusnadi, Vania Clarissa
,
Yulandi, Adi
in
Animals
,
Antibiotic resistance
,
Bacillus cereus
2025
Objectives
Bacteriophages are viruses that specifically target and kill bacteria. Bacteriophages are considered safe for humans, making them suitable for use in food applications Therefore, this study was conducted to isolate, characterize, and apply bacteriophages from the environment to control food spoilage bacteria.
Results
Bacteriophages were isolated by using
Bacillus cereus
(
B. cereus
) as reference host. We found bacteriophage BC-VP titers (1.16 ± 0.18 × 10
9
PFU/mL) and BC-AJ (1.72 ± 0.19 × 10
8
PFU/mL). Both bacteriophages were polyvalent, targeting not only
B. cereus
, but also lysing enteropathogenic
Escherichia coli
(EPEC) and enterotoxigenic
Escherichia coli
(ETEC) with different efficiencies. Bacteriophage BC-VP and BC-AJ miMOI were determined to be 0.01 and 1, respectively. Bacteriophage BC-VP morphology was analyzed using Transmission Electron Microscope (TEM) and categorized as
Myoviridae
family from the order of
Caudovirales
. Both bacteriophages showed significant bacterial reduction when applied to artificially contaminated cooked rice and pasteurized milk. The reduction number was higher in pasteurized milk at 28℃ for BC-VP (93%) and BC-AJ (90%). Molecular characterizations were performed and showed that BC-VP was 96.88% a virulent bacteriophage. Also, no sign of antibiotic resistance gene in BC-VP and genes related to lytic cycle such as putative tail lysin and tail fiber, were annotated.
Journal Article
Determination of bacteriophage ETEC-phage-TG to control pathogenic Escherichia coli
by
Rizkinata, Denny
,
Yulandi, Adi
,
Waturangi, Diana Elizabeth
in
Bacteria
,
Bacteriophage
,
Bacteriophages
2025
Objectives
This study aimed to isolate lytic bacteriophage to control EHEC and ETEC, characterize, and apply their ability to control these two bacteria in food.
Results
One of the bacteria that contaminate the food is Diarrheagenic
Escherichia coli
(DEC) which was consist of Enterohaemorrhagic
E. coli
and Enteropathogenic
E. coli
. Preservation methods were used to improve shelf-life to reduce microbial growth but it might have side effect to the nutrition and human health. Therefore, it is required to explore an alternative method that can be used to be a biocontrol agent for food, including the bacteriophage approach. Our study showed that these two phages are promising to be used as biocontrol agent in food. Genomic DNA sequencing performed that this phage being exclusively lytic.
Journal Article