Influence of lysozyme utilization with lactic acid bacteria in yoghurt on some foodborne pathogens


Published: Nov 9, 2022
Updated: 2022-11-09
Keywords:
Hen Egg White Lysozyme ABT-5 S. aureus clfA E. coli
A Ismail
https://orcid.org/0000-0003-1723-7322
A Deeb
https://orcid.org/0000-0002-2246-0628
I Alhawary
https://orcid.org/0000-0003-1029-0235
W Elkassas
https://orcid.org/0000-0001-5077-1223
Y Hegazy
https://orcid.org/0000-0002-7355-8238
Abstract

Despite the existence of many different food preservation methods, foodborne pathogens are still the most common related problems to dairy products. Developing naturally alternative techniques to control such problems is necessary. This research was planned to assess the bacteriological quality of yoghurt samples commercially available in Kafr El-Sheikh city, Egypt. Also to study the impact of natural antibacterial agents on the inhibition of selected foodborne pathogens artificially inoculated in yoghurt samples. The obtained results revealed poor bacteriological quality of the examined yoghurt samples, as total staphylococci and coliforms were detected in 80% and 90% yoghurt samples, with mean values of 2.4 ×103±0.13× 103cfu/gm and 3.6×105± 2.4×105 MPN/gm, respectively. S. aureus was noticed in 4% of examined samples, polymerase chain reaction (PCR) confirmed the pres­ence of virulence adhesion gene (clfA) in one isolate. E. coli was detected in 14% of yoghurt samples, then serologically identified as O146:H5 (2), O111:H2 (2), O125:H11 (1), O26:H10 (1) and O158:H7 (1). Artificially inoculated yoghurt with S. aureus (7.39 log cfu/gm) and E. coli (8.3 log cfu/gm), were used to investigate the ability of hen egg-white lysozyme either with LAB classic or with ABT-5 (L. acidophilus LA5+ S. thermophiles + Bifidobacterium Bb12) starter cultures to inhibit the growth of these pathogens. During refrigerated storage, all yoghurt batches showed a reduction in pH value. It was observed that both pathogens couldn't be detected in the yoghurt batch containing LAB classic starter + heated lysozyme. Furthermore, S. aureus was completely inhibited in that containing ABT-5 cultures alone, while E. coli decreased by 6.23 log. Conversely, there was not complete inhibition in other batches containing (LAB classic starter, LAB classic starter+ native lysozyme, ABT-5 culture+ native lysozyme and ABT-5 culture+ heated lysozyme) as the reduction rate during all storage periods for S. aureus count was 1.66, 2.63, 4.47  and 4.19 log, While for E. coli count was  3.3, 3.81, 5.53 and 4.89 log,  respectively. This study highlighted the importance of adding natural antibacterial agents like lysozyme with LAB culture to yoghurt as a bio-control strategy to overcome foodborne pathogens that cause a public health hazard.

Article Details
  • Section
  • Research Articles
Downloads
Download data is not yet available.
References
Abd El-Gawad IA, El-Sayed EM, El- Zeini HM, Hafez SA, Saleh FA (2014) Antibacterial activity of probiotic yoghurt and soy-yoghurt against Escherichia coli and Staphylococcus aureus. J Nutr Food Sci 4 (5): 1-6.
Abd El-Halem SG, Attia IA, El-Dera HB, El-Seedy AS (2019) Prevalence and antibiotic resistance of Staphylococcus aureus isolated from raw milk and dairy products collected from Alexandria, Egypt. Alex J Food Sci & Technol 16 (2): 25-33.
Abdou AM, Kim M, Sato K (2013) Functional proteins and peptides of hen’s egg origin. In: Bioactive Food Peptides in Health and Disease. Tech, Croatia-European Union: pp 115–144.
Akram N, Chaudhary AH, Ahmed S, Ghuman MA, Nawaz G, Hussain S (2013) Isolation of bacteria from mastitis affected bovine milk and their Antibiogram. Eur J Vet Med 2 (1): 38-46.
Al-Delaimy KS and Hamamdeh YM (2013) Inhibition of Staphylococcus aureus by lactic acid bacteria and /or Bifidobacterium lactis during milk fermentation and storage. J Microbiol Biotechnol Food Sci 2 (4): 2406-2419.
APHA (American public Health Association) (2004) Standard methods for the examination of Dairy products17th ed. Edited by H. Michael Wehr and Joseph H. Frank, Washington, D.C., USA.
Arabski M, Konieczna I, Tusińska E, Wąsik S, Relich I, Zając K, Kamiński ZJ, Kaca W (2015) The use of lysozyme modified with fluorescein for the detection of Gram-positive bacteria. Microbiol Res 170: 242–247.
Bera A, Biswas R, Herbert S, Kulauzovic E, Weidenmaier C, Peschel A, Go¨tz F (2007) Influence of wall teichoic acid on lysozyme resistance in Staphylococcus aureus. J Bacteriol 189 (1): 280–283.
Buriti FCA, Cardarelli HR, Saad SMI (2007) Biopreservation by Lactobacillus paracasei in co-culture with Streptococcus thermophilus in potentially probiotic and symbiotic fresh cream–cheeses. J Food Prot 70 (1): 228-235.
Cancino-Padilla N, Fellenberg MA, Franco W, Ibáñez RA, Vargas-Bello-Pérez E (2017) Foodborne bacteria in dairy products: Detection by molecular techniques. Cienc Investig Agrar 44 (3): 215-229.
Cardarelli HRI, Saad SMI, Gibson GR, Vulevic J (2007) Functional petit-suisse cheese: measure of the prebiotic effect. Anaerobe, 13 (5-6): 200–207.
Carrillo W, García-Ruiz A, Recio I, Moreno-Arribas MV (2014) Antibacterial activity of hen egg white lysozyme modified by heat and enzymatic treatments against oenological lactic acid bacteria and acetic acid bacteria. J Food Prot 77: 1732–1739.
Cegielska-Radziejewska R and Leśnierowski G, Kijowski J (2009) Antibacterial activity of hen egg white lysozyme modified by thermochemical technique. Eur Food Res Technol 228: 841-845.
El-Kholy M, El-Shinawy SH, Meshref AMS, Korny AM (2014) Screening of antagonistic activity of probiotic bacteria against some food-borne pathogens. J FBT IAU 4 (2): 1-14.
El-Leboudy AA, Amer AA, El-Ansary MA, El- Asuoty MS, Ahmida MR (2017) Evaluation of some fermented milks sold in Alexandria city. Assiut Vet Med J 63 (154): 59-66.
EOSQC (Egyptian Organization for Standardization and Quality Control) (2005): "Domiati cheese 1008-3/Kareish cheese1008-4/ Ice cream 1185-1/Yoghurt 1633".
Erkmen O (1995) Behavior of S. aureus in Turkish feta cheese during manufacture and ripening. J Food Prot 58 (11): 1201-1205.
Fathi SS, Mohamed AS, El- Sayed MSH (2019) Coliforms contamination in raw milk and some dairy products with special reference to comparative identification of Enterobacter spp. Zagazig Vet J 47 (4): 388-397.
Giammanco GM, Pepe A, Aleo A, D’Agostino V, Milone S, Mammina C (2011) Microbiological quality of Pecorino Siciliano “primosale” cheese on retail sale in the street markets of Palermo, Italy. New Microbiol 34 (2): 179-185.
Gorbenko GP, Ioffe VM, Kinnunen PKJ (2007) Binding of lysozyme to phospholipid bilayers: Evidence for protein aggregation upon membrane association. Biophys J 93 (1): 140-153.
Hassan GM, Meshref AMS, Elnewery HA, Mohamed AS (2021) Prevalence of Escherichia coli in milk and some dairy products in Beni-Suef governorate, Egypt. J Vet Med Res 27 (2): 161-167.‏
He M, Yang YX, Han H, Men JH, Bian LH, Wang GD (2005) Effects of yoghurt supplementation on the growth of preschool children in Beijing suburbs. Biomed Environ Sci 18 (3): 192-197.
Heilmann C (2011) Adhesion mechanisms of staphylococci. In: Bacterial Adhesion. Springer, Netherlands: pp 105-123.
Hofi AA and El-Shibiny S (1978) The chemical composition of market yoghurt. Egypt J Dairy Sci 6 (1): 25-31.
Ibrahim HR, Higashiguchi S, Juneja LR, Kim M, Yamamoto T (1996) A structural phase of heat-denatured lysozyme with novel antimicrobial action. J Agric Food Chem 44 (6): 1416–1423.
Ibrahim HR, Matsuzaki T, Aoki T (2001) Genetic evidence that antibacterial activity of lysozyme is independent of its catalytic function. FEBS Lett 506 (1): 27–32.
ISO 4831: 2006 (International Organization for Standardization (2006) Microbiology of food and animal feeding stuffs — Horizontal method for the detection and enumeration of coliforms — Most probable number technique.
ISO 6887-2: 2003(International Organization for Standardization (2003) Microbiology of food and animal feeding stuffs — Preparation of test samples, initial suspension and decimal dilutions for microbiological examination.
ISO 6888-1: 2003 (International Organization for Standardization) (2003) Microbiology of food and animal feeding stuffs — Horizontal methods for the enumeration of coagulase-positive Staphylococci (Staphylococcus aureus and other species) Technique using Baird-Parker medium.
Kandil AA, Elhadidy M, El-Gamal A, Al-Ashmawy M A (2018) Identification of S. aureus and E. coli from dairy products intended for human consumption. Adv Anim Vet Sci 6 (11): 509-513.
Karagozlu N, Karagozlu C, Ergonul B (2007) Survival characteristics of E. coli O157:H7, S. typhimurium and S. aureus during kefir fermentation. Czech J Food Sci 25 (4): 202-207.
Katikou P, Ambrosiadis I, Georgantelis D, Koidis P, Georgakis SA (2005) Effect of Lactobacillus-protective cultures with bacteriocin-like inhibitory substances'producing ability on microbiological, chemical and sensory changes during storage of refrigerated vacuum-packaged sliced beef. J Appl Microbiol 99 (6): 1303-1313.
Kim DH, Chon JW, Kang IB, Kim H, Kim HS, Song KY, Seo KH (2015) Growth inhibition of Cronobacter sakazakii in experimentally contaminated powdered infant formula by kefir supernatant. J Food Prot 78 (9): 1651–1655.
Kok TW, Worswick D, Gowans E (1996) Some serological techniques for microbial and viral infections. In: Practical Medical Microbiology. 14th ed, Edinburgh, Churchill Livingstone, UK: pp 179-204.
L. de Roos A, Walstra P, Geurts TJ (1998) The association of lysozyme with casein. Int Dairy J 8 (4): 319-324.
Leśnierowski G and Cegielska-Radziejewska R (2012) Potential possibilities of production, modification and practical application of lysozyme. Acta Sci Pol Technol Aliment 11 (3): 223–230.
MacFaddin JF (2000) Biochemical tests for identification of medical bacteria. 3rd ed, Lippincott Williams and Wilkin Washington, Washington, Philadelphia, PA 113, U.S.A.
Mann, K. (2017) Proteomics of Egg White. In: Proteomics in Food Science. Kuddus M ed, Academic Press, London, UK: pp 261–276.
Mason WJ, Blevins JS, Beenken K, Wibowo N, Ojha N, Smeltzer MS (2001) Multiplex PCR Protocol for the diagnosis of staphylococcal Infection. J Clin Microbiol 39 (9): 3332–3338.
Masschalck B, Garc´ıa-Graells C, Haver EV, Michiels CW (2000) Inactivation of high pressure resistant Escherichia coli by lysozyme and nisin under high pressure. Innov Food Sci Emerg Technol 1 (1): 39-47.
Meshref AMS, Hassan GM, Riad EM, Ashour WA (2019) Studies on enterotoxigenic Staphylococcus aureus in milk and some dairy products. Assiut Vet Med J 65 (163): 87-97.
Mobarez AM, Hosseini Doust R, Sattari M, Mantheghi N (2008) Antimicrobial effects of bacteriocin like substance produced by L. acidophilus from traditional yoghurt on P. aeruginosa and S. aureus. J Biol Sci 8 (1): 221–224.
Omar SM, Abd El-Fattah HI, Abdelbasit HM, Mahgoub SA (2018) Contamination of fermented foods in Egypt with undesirable bacteria. Zagazig J Agric Res 45 (1): 317-329.
Omola EM, Kawo AH, Shamsudden U (2014) Physico-chemical, sensory and microbiological qualities of yoghurt brands sold in Kano Metropolis, Nigeria. BAJOPAS 7 (2): 26–30.
Quinn PJ, Markey BK, Carter ME, Donnelly WJC, Leonard FC (2002) Veterinary Microbiology and Microbial Diseases. 1st published Blackwell Science, Oxford, UK.
Rada V, Splichal I, Rockova S, Grmanova M, Vlkova E (2010) Susceptibility of bifidobacteria to lysozyme as a possible selection criterion for probiotic bifidobacterial strains. Biotechnol Lett 32(3): 451-455.‏
Robinson RK, Tamine AY, Wszolek M (2000) Microbiology of fermented milks. In: Dairy Microbiology Handbook . 3rd ed, john Wiley and Sons, New York: pp 367-490.
Russo P, Arena MP, Fiocco D, Capozzi V, Drider D, Spano G (2017) Lactobacillus plantarum with broad antifungal activity: a promising approach to increase safety and shelf-life of cereal-based products. Int J Food Microbiol 247: 48–54.
Saleem IM, Gulzar N, Nadeem M, Sameen A, Rafiq S, Rehman S (2021) Influence of hen egg white lysozyme N‐acetylmuramide glycan hydrolase on the antimicrobial activity and textural characteristics of high‐moisture mozzarella cheese. J Food Process Preserv e15573.‏
Sambrook J, Fritscgh EF, Mentiates (1989) Molecular coloning. In: A laboratory manual. Cold spring Harbor Laboratotry press, New York.
Sayed M (2012) Laban rayeb and yoghurt: Sensory, rheological, chemical and microbiological properties. Assiut Vet Med J 58 (132): 1-26.
Sharp JCM, Reilly WJ, Coia JE, Curnow J, Synge BA (1995) Escherichia coli O157:H7 infection in Scotland: an epidemiological overview, 1984-1994. PHLS Microbiol Digest 12:134-140.
Singh P and Prakash A (2008) Isolation of Escherichia coli, Staphylococcus aureus and Listeria monocytogenes from milk products sold under market conditions at Agra Region. Acta agric Slov 92 (1): 83–88.
Soliman NSM and Ahmed LI (2019) Survival of Staphylococcus aureus in Bio-Yoghurt. Open J Appl Sci 9 (7): 564-572.
Tejero- Sariñena S, Barlow J, Costabile A, Gibson GR, Rowland I (2012) In vitro evaluation of the antimicrobial activity of a range of probiotics against pathogens: Evidence for the effects of organic acids. Anaerobe 18 (5): 530-538.
Thammasirirak S, Pukcothanung Y, Preecharram S, Daduang S, Patramanon R, Fukamizo T, Araki T (2010) Antimicrobial peptides derived from goose egg white lysozyme. Comp Biochem Physiol C Toxicol Pharmacol 151 (1): 84–91.
Touch V, Hayakawa S, Saitoh K (2004) Relationships between conformational changes and antimicrobial activity of lysozyme upon reduction of its disulfides bonds. J Food Chem 84 (3): 421–428.
Turchia B, Pedonesea F, Torraccaa B, Fratinia F, Mancinia S, Galieroa A, Montalbanoa B, Cerria D, Nuvolonia R (2017) Lactobacillus plantarum and Streptococcus thermophilus as starter cultures for a donkey milk fermented beverage. Int J Food Microbiol 256 (1): 54–61.
Valilis E, Ramsey A, Sidiq S, DuPont HL (2018) Non-O157 Shiga toxin-producing Escherichia coli-A poorly appreciated enteric pathogen: Systematic review. Int J Infect Dis 76: 82–87.
Vaudaux PE, Francois P, Proctor RA, McDevitt D, Foster TJ, Albrecht RM, Lew DP, Wabers H, Cooper SL (1995) Use of adhesion-defective mutants of Staphylococcus aureus to define the role of specific plasma proteins in promoting bacterial adhesion to canine arteriovenous shunts. Infect Immun 63 (2): 585-590.‏
Vilcacundo R, Méndez P, Reyes W, Romero H, Pinto A, Carrillo W (2018) Antibacterial activity of hen egg white lysozyme denatured by thermal and chemical treatments. Sci Pharm 86 (48): 1-17.
Vinderola CG, Costa GA, Regenhardt S, Reinheimer JA (2002) Influence of compounds associated with fermented dairy products on the growth of lactic acid starter and probiotic bacteria. Int Dairy J 12 (7): 579–589.
Yabaya A and Idris A (2012) Bacteriological quality assessment of some yoghurt brands sold in Kaduna Metropolis. JORIND 10 (2):35-39.
Yesillik S, Yildirim N, Dikici A, Yildiz A, Yesillik S (2011) Antibacterial effects of some fermented commercial and homemade dairy products and 0.9% lactic acid against selected foodborne pathogens. Asian J Anim Vet Adv 6 (2): 189-195.
Younis MO, Baka ZAM, Abo-Doubara MI, El-Metwaly MAM, Mostafa AHM (2020) Utilization of Cyanobacteria extracts in improving the microbial quality of soft white cheese. J Agric Chem Biotechnol 11 (7): 223-228.‏
Most read articles by the same author(s)