Bioprotection in Vegetable Foods Antonio Glvez Prof. of - - PowerPoint PPT Presentation

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Bioprotection in Vegetable Foods Antonio Glvez Prof. of - - PowerPoint PPT Presentation

Jun 20, 2023 468 likes •682 views

Bioprotection in Vegetable Foods Antonio Glvez Prof. of Microbiology Dept.of Health Sciences University of Jaen, Spain agalvez@ujaen.es Microorganisms in vegetable foods Cross Raw materials Processing contamination Pathogens Toxin

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Bioprotection in Vegetable Foods

Antonio Gálvez

  • Prof. of Microbiology

Dept.of Health Sciences University of Jaen, Spain agalvez@ujaen.es

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FOOD Pathogens Spoilage

Beneficial effects

Toxin producers Processing Raw materials Cross contamination

Microorganisms in vegetable foods

Antimicrobial resistance

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Hydroxy-fatty acids Cyclic dipeptides Antifungal peptides Carboxylic acids Organic acids Aldehydes Organic acids Aldehydes Bacteriocins Miscellaneous Tetramic acid ANTIBACTERIAL ANTIFUNGAL

ANTIMICROBIAL SUBSTANCES FROM LAB

Synergistic action

LAB

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Lactobacillus

  • Lb. acidophilus
  • Lb. amylovorus
  • Lb. brevis
  • Lb. coryniformis
  • Lb. hammesii
  • Lb. paracollinoides
  • Lb. pentosus
  • Lb. plantarum
  • Lb. reuteri
  • Lb. sakei
  • Lb. sanfranciscensis

Lactococcus

  • L. lactis

Leuconostoc

  • Ln. citreum
  • Ln. mesenteroides

Pediococcus

  • P. acidilactici
  • P. pentosaceous

Weissella

  • W. cibaria
  • W. confusa
  • W. paramesenteroides

ANTIFUNGAL LAB FROM VEGETABLE MATERIALS AND FOODS

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ANTIFUNGAL LAB AS BIOPROTECTIVE CULTURES Inhibition of spoilage and mycotoxigenic fungi

Fresh fruits and vegetables Sourdoughs and breads Fruit juices Rice cakes Fermented beverages

Prevention of Korean rice wine spoilage (Ryu et al. http://dx.doi.org/10.1016/j.fm.2014.01.011)

Treated Control

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POTENTIAL OF BACTERIOCINOGENIC LAB IN THE PRESERVATION OF VEGETABLES AND FRUITS Bioprotective cultures for fruits and vegetables

  • Increased safety of minimally processed vegetables due to the inhibition
  • f A. hydrophila, Staph. aureus, E. coli and L. monocytogenes
  • Growth inhibition of E. coli and S. typhimurium and inactivation of L.

monocytogenes on wounded Golden Delicious apples and Iceberg lettuce cut leaves

  • Increased safety and shelf life of lamb´s lettuce and apple cubes
  • Improving the safety and shelf life of fruits and vegetables by nisin-

producing strains

Starter cultures in vegetable fermentations

  • Avoiding stuck fermentations
  • Acceleration of fermentation
  • To improve the consistency and quality of the final products
  • Protection from undesirable spoilage and pathogenic microorganisms
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Plantaricin genes detected in L. pentosus strains

plnA plnD plnJ plnNC8 plnW

Fermented table olives as sources of strains producing antimicrobial substances

Antimicrobial activity of LAB from table

  • lives (Abriouel et al., 2012 (doi:

10.1016/j.fm.2012.07.006)

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POTENTIAL OF LAB BACTERIOCINS IN PRESERVATION OF FRESH VEGETABLES AND FRUITS

Nisin

  • Reducing the surface bacterial load on whole melon and transfer of

pathogens from the surface of melons to freshly cut pieces

  • Reducing the load of pathogens on the surface of minimally

processed mangoes (nisin film) Pediocin

  • Preserving minimally processed papaya (alginate coatings)

Pentocin MQ1

  • Extension of the shelf life of bananas

Enterocin 416K1

  • Growth inhibition of L. monocytogenes on

apples and grapes

Prevention of banana spoilage (Wayah & Philip, https://doi.org/10.3389/fmicb.2018.00564)

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Enterocin AS-48

Circular bacteriocin Wide bactericidal spectrum

  • Bacillus
  • Alicyclobacillus
  • Clostridium
  • Listeria monocytogenes
  • Staphylococcus aureus
  • Lactobacillus
  • Pediococcus
  • Escherichia coli*
  • Salmonella*
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ENTEROCIN AS-48 FOR BIOPROTECTION OF VEGETABLES

Fresh vegetables Fruits Cereals Fermented vegetables

AS-48

Soups, purees and sauces Bakery products Canned vegetables Beverages

Chemical preservatives Essential oils Tª, HIPEF, HHP

Synergy with

  • ther

hurdles

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  • Applications in fermented foods
  • Manzanilla Aloreña table olives
  • Short fermentation during cold storage

(ca. one week)

  • Packed in brine with condiments
  • Do not withstand pasteurization
  • Short shelf life due to refermentation

HHP

Bacteriocins (nisin, AS-48) Essential oils

No refermentation at room temperature Reduced salt content

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  • Fermented beverages

Endospores of B. licheniformis were more resistant to AS-48. Endospore inactivation improved in combination with mild heat treatment

Final bacteriocin concentrations were 2 ( ▲ ), 4 ( ● ), and 10 µg/ml (□ ). Controls (Ο ).

2 4 6 5 10 15

Time (days) Log CFU/ml

4ºC

Bacillus licheniformis

Inactivation of spoilage bacteria in beer (except hop-resistant strains) Inactivation of spoilage bacteria in white wine (but not in red wine) Inactivation of spoilage lactic acid bacteria and endospore formers in apple ciders

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  • Fruit juices

A B

Vegetative cells as well as endospores of A. acidoterrestris are highly sensitive to AS-48 (2.5 µg/ml at 4 and 15ºC) in commercial and fresh- made juices Inactivation of S. enterica improved in combination with HiPEF

Alicyclobacillus acidoterrestris

AS-48 (30-60 µg/ml)

Salmonella enterica

2.75 3 3.25 3.5 3.75 4 4.25 4.5

AS-48 (µg/ml)

30 37.5 45 52.5 60

Treatment time (µs)

100 325 550 775 1000

40ºC

HiPEF

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1 2 3 4 5 6

AS-48 + Hypochlorite (100 ppm) AS-48 + Hypochlorite (50 ppm) AS-48 + Hypochlorite (25 ppm) AS-48 + HDP (0.1%) AS-48 + Peracetic acid (80 ppm) AS-48 + PHBME (0.5%) AS-48 + PHBME (0.1%) AS-48 + Propyl-p-HB (0.5%) AS-48 + Propy-p-HB (0.1%) AS-48 + Permanganate (25 ppm) AS-48 + Thiosulfate (0.01 N) AS-48 + TSTMP (0.5%) AS-48 + TSTMP (0.1%) AS-48 + TSP (1.5%) AS-48 + Nitrate (100 ppm) AS-48 + Nitrate (50 ppm) AS-48 + Nitrite (100 ppm) AS-48 + Nitrite (50 ppm) AS-48 + Lactate (0.5%) AS-48 + Lactate (0.1%) AS-48 + Lactic acid (0.5%) AS-48 + Lactic acid (0.1%) AS-48 + Sorbate (0.1%) AS-48 + Propionate (0.5%) AS-48 + Propionate (0.1%) AS-48 + Citric acid (0.5%) AS-48 + Acetic acid (0.5%) AS-48 Control

Treatment Log CFU/ml

Enterocin AS-48 reduced viable cell counts in fresh produce (sprouts, sliced fruits). Strong synergistic activity with several chemical preservatives, essential

  • ils,

and phenolic compounds.

  • L. monocytogenes
  • B. cereus
  • B. weinhestephanensis

Control of pathogenic and spoilage bacteria in fresh produce

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  • Salmonella enterica
  • Escherichia coli
  • Aeromonas hydrophila
  • Shigella sonnei
  • Enterobacter aerogenes
  • Yersinia enterocolitica
  • Pseudomonas fluorescens

AS-48 Control Lactic acid + AS-48 Lactic acid (1.5%) Trisodium P + AS-48 Trisodium P (1.5%) PolyP + AS-48 PolyP (0.1%) HDP + AS-48 HDP (0.5%) Peracetic acid + AS-48 Peracetic acid (80 ppm)

1 2 3 4 5 6

Inactivation of Gram-negative bacteria in sprouts combination with other antimicrobials

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2 4 6 1 3 7

Log10 CFU/g Time (days)

2 4 6 1 2 3 4 5 6 7

Log10 CFU/g Time (days)

Activated coatings

  • Chitosan
  • Caseinate
  • Alginate
  • K-carrageenate
  • Xanthan gum
  • Pectin
  • Starch
  • Carboxymethyl cellulose
  • Methyl cellulose

Pectin Pectin + EDTA

Inactivation of Listeria monocytogenes in apple cubes

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0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100%

Proteobacteria Bacteroidetes Firmicutes Actinobacteria

0% 10% 20% 30% 40% 50% 60% 70% 80% 90% 100% C0 C3 C5 C10 PB0 PB3 PB5 PB10

Pseudomonas Gamma Proteobacterium Rheinheimera Flavobacterium Acinetobacter Shigella Stenotrophomonas Salmonella

  • Unc. low G+C G+

Paenibacillus Sphingobacterium Erwinia Pantoea Rest

Effect of pectin-bacteriocin coating in chopped parsley

Viable counts (Log10 CFU/g ± SD) at different storage times (days) Treatment 3 7 10 Control 6.3 ± 0.05 6.8 ± 0.09 7.8 ± 0.09 9.4 ± 0.06 PB 2.6 ± 0.07 2.3 ± 0.42 3.4 ± 0.24 4.0 ± 0.52

Relative abundance (%) Relative abundance (%)

Grande et al., 2017. doi: 10.1016/j.foodres.2017.05.011.

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Relative abundance (%)

Effect on bacterial diversity of cherimoya pulp

Perez Pulido et al.2015. doi: 10.1016/j.ijfoodmicro.2014.11.033.

10 20 30 40 50 60 70 80 90 100

Pantoea vagans Pantoea agglomerans Pantoea ananatis Enterobacter aerogenes Enterobacter asburiae Enterobacter kobei Escherichia fergusonii Leclercia adecarboxylata Raoultella terrigena Serratia fonticola Serratia liquefaciens Serratia plymuthica Serratia proteamaculans Erwinia aphidicola Erwinia billingiae Erwinia persicina Yersinia ruckeri Cronobacter turicensis Acinetobacter johnsonii Pseudomonas psychrophila Pseudomonas putida Pseudomonas fragi Stenotrophomonas rhizophila Enterococcus casseliflavus Enterococcus gallinarum Leuconostoc kimchii Leuconostoc mesenteroides Leuconostoc sp. C2 Bacillus firmus Bacillus stratosphericus Bacillus plakortidis Bacillus nealsonii Bacillus pumilus Bacillus thuringiensis Micrococcus luteus

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Thank you for your attention