Presence of methicillin-resistant Staphylococcus aureus in raw cow’s milk: adhesive capacities and extracellular enzymes characterization

PDF (English)
Δημοσιευμένα: oct. 18, 2023
Εκδόσεις:
2023-10-18 (1)
DOI: https://doi.org/10.12681/jhvms.31440
AR Belmamoun
Department of Agricultural Sciences, Faculty of Nature and Life Sciences, DjillaliLiabes University, Sidi- Bel-Abbes, Algeria
A Ammam
Laboratory of Pharmacognosy, Bio toxicology, and Biological Valorization of Plants, MoulayTahar University, BP 20000, Saida, Algeria
C Mhamdia
Department of environmental sciences, faculty of natural and life sciences, Djilali Liabes University, BP 22000, Sidi-Bel Abbès, Algeria
R Chadli
Laboratory of Organic and Macromolecular Physical Chemistry, Djillali Liabes University, BP 22000, Sidi-Bel-Abbés, Algeria
A Baki
Department of environmental sciences, faculty of natural and life sciences, Djilali Liabes University, BP 22000, Sidi-Bel Abbès, Algeria
N Madouni
Department of Agricultural Sciences, Faculty of Nature and Life Sciences, Djillali Liabes University, BP 22000, Sidi-Bel-Abbes
Περίληψη

Raw milk contamination by Methicillin-resistant S. aureus (MRSA) threatens food safety and leads to public health problems. Our study evaluates biofilm formation and virulence factors among 21 MRSA in raw cow’s milk. Methicillin resistance was confirmed by cefoxitin screening using the automated VITEK2 system, with a minimum inhibitory concentration greater than 8 mg/l. Qualitative characterization of biofilm-producing strains was performed on Congo red agar (CRA), with a semi-quantitative adhesion test on 96-well tissue culture plates (TCP). The ability to produce different enzymes was evaluated, such as caseinase, lipase, and phospholipase (lecithinase). The surface hydrophobicity of the bacteria was determined, and the autoaggregation test was used to predict the interactions between bacterial cells. Among the tested strains, 61.9% were biofilm producers in the CRA, developing a positive and variable phenotype. Furthermore, 19.05% and 80.95% of isolates were high and low biofilm formation on TCP. The enzymatic activity showed that lécithinase, caseinase, and lipase activities were detected in 100%, 80.95%, and 80.95% of cases. Highly hydrophilic (85.71%) and weakly hydrophobic (14.29%) were detected in MRSA isolates. The strains showed that 71.43% were moderate autoaggregation and 28.57% were low autoaggregation. No significant difference was found between the CRA method and TCP (P> 0.05). A significant association was found between adhesion capacity and bacterial autoaggregation in S. aureus strains (p>0.05). On the other hand, no statistical association between the hydrophobicity of microbial strains and adhesion capacity (p<0.05) was found. The same result was for the hydrophobicity of microbial strains and autoaggregation (p<0.05). This investigation could be beneficial for developing new control measures, prevention, and effective treatment against staphylococcal bacterial resistance.

Λεπτομέρειες άρθρου
  • Ενότητα
  • Research Articles
Creative Commons License

Αυτή η εργασία είναι αδειοδοτημένη υπό το CC Αναφορά Δημιουργού – Μη Εμπορική Χρήση 4.0.

Οι συγγραφείς των άρθρων που δημοσιεύονται στο περιοδικό διατηρούν τα δικαιώματα πνευματικής ιδιοκτησίας επί των άρθρων τους, δίνοντας στο περιοδικό το δικαίωμα της πρώτης δημοσίευσης.

Άρθρα που δημοσιεύονται στο περιοδικό διατίθενται με άδεια Creative Commons 4.0 Non Commercial και σύμφωνα με την άδεια μπορούν να χρησιμοποιούνται ελεύθερα, με αναφορά στο/στη συγγραφέα και στην πρώτη δημοσίευση για μη κερδοσκοπικούς σκοπούς.

Οι συγγραφείς μπορούν να καταθέσουν το άρθρο σε ιδρυματικό ή άλλο αποθετήριο ή/και να το δημοσιεύσουν σε άλλη έκδοση, με υποχρεωτική την αναφορά πρώτης δημοσίευσης στο J Hellenic Vet Med Soc

Οι συγγραφείς ενθαρρύνονται να καταθέσουν σε αποθετήριο ή να δημοσιεύσουν την εργασία τους στο διαδίκτυο πριν ή κατά τη διαδικασία υποβολής και αξιολόγησής της.

Λήψεις
Τα δεδομένα λήψης δεν είναι ακόμη διαθέσιμα.
Αναφορές
Achek R, Hotzel H, Nabi I, Kechida S, Mami D, Didouh N, Tomaso H,
Neubauer H, EhrichtR, MoneckeS (2020) Phenotypic and molecular
detection of biofilm formation in Staphylococcus aureus isolated from
different sources in Algeria. Pathogens 9 (2): 153. Doi :1 0. 3390/patho
gens9020153
Ahmed W, Neubauer H, Tomaso H, El Hofy FI, Monecke S, Abdeltawab
AA, Hotzel H (2020) Characterization of Staphylococci and Strepto
cocci Isolated from Milk of Bovides with Mastitis in Egypt. Pathogens
(5): 381. Doi :10. 3390/pathogens9050381
Atulya M, Mathew AJ, Rao JV, Rao CM (2014) Influence of milk compo
nents in establishing biofilm mediated bacterial mastitis infections in
cattle: a fractional factorial approach. Res Vet Sci96 (1): 25-27. Doi :10.
/j. rvsc. 2013. 12. 001
Auger S, Ramarao N, Faille C, Fouet A, Aymerich S, Gohar M (2009)
Biofilm formation and cell surface properties among pathogenic and
nonpathogenic strains of the Bacillus cereus group. Appl Environ Mi
crobiol 75 (20) :6616-6618. Doi :10. 1128/aem. 00155-09
Belkadi S, Meradi M, Oucheriah Y, Yahiaoui FE, Bouakaz A, Nezzar KW,
Heleili N (2022)Occurrence of Methicillin Resistant Staphylococcus
aureus Carriage in Different Animal Species in Eastern Algeria. Agri
cultural Science Digest42 (2): 217-222. Doi :10. 18805/ag. d-329
BelmamounAR, Ammam A, Berrabah I, Reguig KB (2017) Coagulase gene
polymorphism and antimicrobial susceptibility of Staphylococcus au
reus isolated from bovine subclinical mastitis milk in Sidi-Bel-Abbes,
Algeria. South Asian J Exp Biol 7 (1):21–27. Doi :10. 38150/sajeb. 7
(1). p21-27
Borges MT, Nascimento AG, RochaUN, Tótola MR (2008) Nitrogen star
vation affects bacterial adhesion to soil. Braz J Microbiol. 39 (3):457
Doi:10. 1590/s1517-83822008000300009
Canning B, Mohamed I, Wickramasinghe N, Swindells J, O’Shea MK
(2020)Thermonuclease test accuracy is preserved in methicillin-resis
tant Staphylococcus aureus isolates. J Med Microbiol69 (4):548-551.
Doi :10. 1099/jmm. 0. 001166
Cha JO, Park YK, Lee YS, Chung GT (2011) In vitro biofilm formation and
bactericidal activities of methicillin-resistant Staphylococcus aureus
clones prevalent in Korea. DiagnMicrobiol Infect Dis70 (1):112-118. Doi :10. 1016/j. diagmicrobio. 2010. 11. 018
Chen L, Tang ZY, Cui SY, Ma ZB, Deng H, Kong WL, Yang LW, Lin C,
Xiong WG, Zeng ZL (2020)Biofilm Production Ability, Virulence and
Antimicrobial Resistance Genes in Staphylococcus aureus from Var
ious Veterinary Hospitals. Pathogens9 (4):264. Doi : 10. 3390/patho
gens9040264
Chen X, Alonzo F (2019) Bacterial lipolysis of immune-activating ligands
promotes evasion of innate defenses. Proc Natl Acad Sci 116 (9):3764
Doi :10. 1073/pnas. 1817248116
CLSI (2008) Performance standards for antimicrobial disk and dilution sus
ceptibility tests for bacteria isolated from animals, 3rd Ed. Wayne, PA:
Approved standard, M31-A3
Collado MC, Surono I, MeriluotoJ, Salminen S (2007) Indigenous dadih
lactic acid bacteria: cell‐surface properties and interactions with patho
gens. J Food Sci72 (3): M89-M93. Doi :10. 1111/j. 1750-3841. 2007.
x
Darwish SF, Asfour HA (2013) Investigation of biofilm forming ability in
Staphylococci causing bovine mastitis using phenotypic and genotypic
assays. Sci World J 378492:1-9. Doi :10. 1155/2013/378492
Del Re B, Sgorbati B, Miglioli M, Palenzona D (2000) Adhesion, autoag
gregation and hydrophobicity of 13 strains of Bifidobacterium longum.
Lett Appl Microbiol31 (6):438-442. Doi :10. 1046/j. 1365-2672. 2000.
x
Dhanawade NB, Kalorey DR, Srinivasan R, Barbuddhe SB, Kurkure NV
(2010) Detection of intercellular adhesion genes and biofilm produc
tion in Staphylococcus aureus isolated from bovine subclinical masti
tis. Vet Res Commun34 (1):81-89. Doi :10. 1007/s11259-009-9326-0
El-Jakee J, Nagwa AS, Bakry M, Zouelfakar SA, ElgabryE, El-Said WG
(2008) Characteristics of Staphylococcus aureus strains isolated from
human and animal sources. Am. -Eurasian jagric environ sci4 (2):221
Gayatri S, DeepakS, Hardik G (2017) Biofilm formation, hemolysin pro
duction and antimicrobial susceptibilities of Staphylococcus aureus
isolated from the mastitis milk of buffaloes in Udaipur, India. Int J Vet
Sci 6 (1):1-6.
Haddad O, Merghni A, Elargoubi A, Rhim H, Kadri Y, Mastouri M (2018)
Comparative study of virulence factors among methicillin resistant
Staphylococcus aureus clinical isolates. BMC Infect Dis18 (1):560.
Doi :10. 1186/s12879-018-3457-2
Hoveida L, Halaji M, Rostami S, Mobasherizadeh S (2019) Biofilm-pro
ducing ability of Staphylococcus spp isolated from different foodstuff
products. Ann Ig31:140-147. Doi :10. 7416/ai. 2019. 226
Joo HS, Chatterjee SS, Villaruz AE, Dickey SW, Tan VY, Chen Y, Stur
devant DE, Ricklefs SM, Otto M (2016) Mechanism of gene regula
tion by a Staphylococcus aureus toxin. MBio7 (5): e01579-16. Doi :10.
/mbio. 01579-16
Kanjan P, Sakpetch P (2020) Functional and safety assessment of Staphylo
coccus simulans PMRS35 with high lipase activity isolated from high
salt-fermented fish (Budu) for starter development. LWT 124:109183.
Doi: 10. 1016/j. lwt. 2020. 109183
Komodromos D, Kotzamanidis C, Giantzi V, Angelidis AS, Zdragas A,
Sergelidis D (2022) Prevalence and biofilm-formation ability of Staph
ylococcus aureus isolated from livestock, carcasses, the environment,
and workers of three abattoirs in Greece. J Hellenic Vet Med Soc73
(2):4097–4104. Doi :10. 12681/jhvms. 26469
Kot B, Szweda P, Frankowska-Maciejewska A, Piechota M, Wolska K
(2016) Virulence gene profiles in Staphylococcus aureus isolated
from cows with subclinical mastitis in eastern Poland. J Dairy Res 83
(2):228-235. Doi :10. 1017/s002202991600008x
Li Q, Liu X, Dong M, Zhou J, Wang Y (2015) Aggregation and adhesion
abilities of 18 lactic acid bacteria strains isolated from traditional
fermented food. Int J Agric Policy Res 3 (2):84-92. Doi :10. 15739/
IJAPR. 030
Mack D, Bartscht K, Fischer C, Rohde H, de Grahl C, Dobinsky S, Horst
kotte MA, Kiel K, Knobloch JKM (2001)Genetic and biochemical
analysis of Staphylococcus epidermidis biofilm accumulation. Methods
Enzymol 336: 215–239. Doi:10. 1016/s0076-6879 (01)36592-8
Markey B, Leonard F, Archambault M, Cullinane A, Maguire D (2013)
Clinical Veterinary Microbiology. 2nded, Mosby Elsevier, China: pp
-119.
Marques VF, Motta CCd, Soares Bds, MeloDAd, Coelho SdMdO, Coel
ho Ids, Barbosa HS, Souza MMSd (2017) Biofilm production and be
ta-lactamic resistance in Brazilian Staphylococcus aureus isolates from
bovine mastitis. Braz J Microbiol48 (1):118-124. Doi :10. 1016/j. bjm.
10. 001
Marques VF, Souza M, de Mendonça EC, AlencarTAd, Pribul BR, Coel
ho Sdmdo, Lasagno M, Reinoso EB (2013) Phenotypic and genotypic
analysis of virulence in Staphylococcus spp. and its clonal dispersion
as a contribution to the study of bovine mastitis. Pesqui Vet Bras 33
(2):161-170. Doi :10. 1590/s0100-736x2013000200005
Melo PdC, Ferreira LM, Nader Filho A, Zafalon LF, Vicente HIG, Souza Vd
(2013) Comparison of methods for the detection of biofilm formation by
Staphylococcus aureus isolated from bovine subclinical mastitis. Braz J
Microbiol 44 (1):119-124. Doi : 10. 1590/s1517-83822013005000031
Merghni A, Nejma MB, Hentati H, Mahjoub A, Mastouri M (2014) Adhe
sive properties and extracellular enzymatic activity of Staphylococcus
aureus strains isolated from oral cavity. Microb. Pathog 73:7-12. Doi
:10. 1016/j. micpath. 2014. 05. 002
Papadopoulos T, Komodromos D, Papadopoulos P, Malissiova E,
Torounidou P, Chouliara E, Zdragas A, Chaligiannis I, Pardali D,
Sergelidis D (2021) Nasal carriage of Staphylococcus aureus among
healthy veterinary students in Greece, 2017-2018: A cross-sectional co
hort study. J Hellenic Vet Med Soc 72 (2): 2833–2842. Doi:10. 12681/
jhvms. 27520
Parth F, Chauhan H, Bhagat A, Chandel B, Dadawala A, Kher H (2016)
Detection of virulence associated factors from Staphylococcus aureus
isolated from bovine mastitis. Buffalo Bull35 (4):687-696.
Rahman MM, Kim WS, Kumura H, Shimazaki Ki (2008)Autoaggregation
and surface hydrophobicity of bifidobacteria. World J MicrobiolBio
technol24 (8):1593-1598. Doi :10. 1007/s11274-007-9650-x
Raksha L, Gangashettappa N, Shantala G, Nandan BR, Sinha D (2020)
Study of biofilm formation in bacterial isolates from contact lens wear
ers. Indian J Ophthalmol 68 (1):23-28. Doi :10. 4103/ijo. ijo_947_19
Ren Q, Liao G, WuZ, Lv J, Chen W (2020) Prevalence and characteriza
tion of Staphylococcus aureus isolates from subclinical bovine mastitis
in southern Xinjiang, China. J Dairy Sci103 (4):3368-3380. Doi :10.
/jds. 2019-17420
Rodríguez-Lázaro D, Alonso-Calleja C, Oniciuc EA, Capita R, Gallego D,
González-Machado C, Wagner M, Barbu V, Eiros-Bouza JM, Nicolau
AI (2018) Characterization of biofilms formed by foodborne methicil
lin-resistant Staphylococcus aureus. Front Microbiol9:3004. Doi :10.
/fmicb. 2018. 03004
Sharma V, Sharma S, Dahiya DK, Khan A, Mathur M, Sharma A (2017)
Coagulase gene polymorphism, enterotoxigenecity, biofilm production,
and antibiotic resistance in Staphylococcus aureus isolated from bo
vine raw milk in North West India. Ann Clin MicrobiolAntimicrob 16
(1):65. Doi :10. 1186/s12941-017-0242-9
Shivaee A, Kalani BS, Talebi M, Darban-Sarokhalil D (2019) Does biofilm
formation have different pathways in Staphylococcus aureus? Iran J
Basic Med Sci 22 (10):1147-1152. Doi :10. 22038/ijbms. 2019. 34888.
Tam K, Torres VJ (2019)Staphylococcus aureus Secreted Toxins and Extra
cellular Enzymes. MicrobiolSpectr7 (2). Doi :10. 1128/9781683670131.
ch40
TorlakE, Korkut E, Uncu AT, Şener Y (2017) Biofilm formation by Staphy
lococcus aureus isolates from a dental clinic in Konya, Turkey. J Infect
Public Health10 (6):809-813. Doi :10. 1016/j. jiph. 2017. 01. 004
TouatiA, Achour W, Abbassi M, Ben AH (2007) Detection of ica genes and
slime production in a collection of Staphylococcus epidermidis strains
from catheter-related infections in neutropenic patients. Pathol Biol
(Paris)55 (6) :277-282. Doi :10. 1016/j. patbio. 2007. 03. 003
Trunk T, Khalil HS, Leo JC (2018) Bacterial autoaggregation. AIMS micro
biol 4 (1):140-164. Doi :10. 3934/microbiol. 2018. 1. 140
Vasil M, Farkasova Z, Elecˇko J, Illek J, Zigo F (2017) Comparison of bio
f
ilm formation by Staphylococcus aureus and Staphylococcus epider
midis strains isolated from sheep milk using three diagnostic methods.
Pol J Vet Sci20 (4):795-801. Doi :10. 1515/pjvs-2017-0100
Waśko A, Polak-Berecka M, Paduch R, Jóźwiak K (2014) The effect of
moonlighting proteins on the adhesion and aggregation ability of Lac
tobacillus helveticus. Anaerobe 30:161-168. Doi :10. 1016/j. anaerobe.
10. 002
Wu Y, Li J, Qiao M, Meng D, Meng Q, Qiao J, Zhang X, Wang L, Cai K,
Zhang J (2019)Characteristic profiles of biofilm, enterotoxins and vir
ulence of Staphylococcus aureus isolates from dairy cows in Xinjiang
Province, China. J Vet Sci 20 (6):e74. Doi : 10. 4142/jvs. 2019. 20. e74
Xu H, Jeong H, Lee H, Ahn J (2009) Assessment of cell surface properties
and adhesion potential of selected probiotic strains. Lett Appl Microbi
ol49 (4):434-442. Doi :10. 1111/j. 1472-765x. 2009. 02684. x
Yong YY, Dykes GA, Choo WS (2019) Biofilm formation by staphylococci
in health-related environments and recent reports on their control us
ing natural compounds. Crit Rev Microbiol 45 (2):201-222. Doi :10.
/1040841x. 2019. 1573802
Zaatout N, Ayachi A, Kecha M (2020) Epidemiological investigation of
subclinical bovine mastitis in Algeria and molecular characterization
of biofilm-forming Staphylococcus aureus. Trop Anim Health Prod52
(1):283-292. Doi :10. 1007/s11250-019-02015-9