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      Staphylococcus aureus Entrance into the Dairy Chain: Tracking S. aureus from Dairy Cow to Cheese

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          Abstract

          Staphylococcus aureus is one of the most important contagious mastitis pathogens in dairy cattle. Due to its zoonotic potential, control of S. aureus is not only of great economic importance in the dairy industry but also a significant public health concern. The aim of this study was to decipher the potential of bovine udder associated S. aureus as reservoir for S. aureus contamination in dairy production and processing. From 18 farms, delivering their milk to an alpine dairy plant for the production of smeared semi-hard and hard cheese. one thousand hundred seventy six one thousand hundred seventy six quarter milk (QM) samples of all cows in lactation ( n = 294) and representative samples form bulk tank milk (BTM) of all farms were surveyed for coagulase positive (CPS) and coagulase negative Staphylococci (CNS). Furthermore, samples from different steps of the cheese manufacturing process were tested for CPS and CNS. As revealed by chemometric-assisted FTIR spectroscopy and molecular subtyping ( spa typing and multi locus sequence typing), dairy cattle represent indeed an important, yet underreported, entrance point of S. aureus into the dairy chain. Our data clearly show that certain S. aureus subtypes are present in primary production as well as in the cheese processing at the dairy plant. However, although a considerable diversity of S. aureus subtypes was observed in QM and BTM at the farms, only certain S. aureus subtypes were able to enter and persist in the cheese manufacturing at the dairy plant and could be isolated from cheese until day 14 of ripening. Farm strains belonging to the FTIR cluster B1 and B3, which show genetic characteristics (t2953, ST8, enterotoxin profile: sea/ sed/ sej) of the recently described S. aureus genotype B, most successfully contaminated the cheese production at the dairy plant. Thus, our study fosters the hypothesis that genotype B S. aureus represent a specific challenge in control of S. aureus in the dairy chain that requires effective clearance strategies and hygienic measures already in primary production to avoid a potential transfer of enterotoxic strains or enterotoxins into the dairy processing and the final retail product.

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          Staphylococcal enterotoxins.

          Staphylococcus aureus is a major human pathogen that produces a wide array of toxins, thus causing various types of disease symptoms. Staphylococcal enterotoxins (SEs), a family of nine major serological types of heat stable enterotoxins, are a leading cause of gastroenteritis resulting from consumption of contaminated food. In addition, SEs are powerful superantigens that stimulate non-specific T-cell proliferation. SEs share close phylogenetic relationships, with similar structures and activities. Here we review the structure and function of each known enterotoxin.
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            Microbiological characterizations by FT-IR spectroscopy.

            Infrared signals of microorganisms are highly specific fingerprint-like patterns that can be used for probing the identity of microorganisms. The simplicity and versatility of Fourier-transform infrared spectroscopy (FT-IR) makes it a versatile technique for rapid differentiation, classification, identification and large-scale screening at the subspecies level.
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              Molecular Correlates of Host Specialization in Staphylococcus aureus

              Background The majority of Staphylococcus aureus isolates that are recovered from either serious infections in humans or from mastitis in cattle represent genetically distinct sets of clonal groups. Moreover, population genetic analyses have provided strong evidence of host specialization among S. aureus clonal groups associated with human and ruminant infection. However, the molecular basis of host specialization in S. aureus is not understood. Methodology/Principal Findings We sequenced the genome of strain ET3-1, a representative isolate of a common bovine mastitis-causing S. aureus clone. Strain ET3-1 encodes several genomic elements that have not been previously identified in S. aureus, including homologs of virulence factors from other Gram-positive pathogens. Relative to the other sequenced S. aureus associated with human infection, allelic variation in ET3-1 was high among virulence and surface-associated genes involved in host colonization, toxin production, iron metabolism, antibiotic resistance, and gene regulation. Interestingly, a number of well-characterized S. aureus virulence factors, including protein A and clumping factor A, exist as pseudogenes in ET3-1. Whole-genome DNA microarray hybridization revealed considerable similarity in the gene content of highly successful S. aureus clones associated with bovine mastitis, but not among those clones that are only infrequently recovered from bovine hosts. Conclusions/Significance Whole genome sequencing and comparative genomic analyses revealed a set of molecular genetic features that distinguish clones of highly successful bovine-associated S. aureus optimized for mastitis pathogenesis in cattle from those that infect human hosts or are only infrequently recovered from bovine sources. Further, the results suggest that modern bovine specialist clones diverged from a common ancestor resembling human-associated S. aureus clones through a combination of foreign DNA acquisition and gene decay.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                13 October 2016
                2016
                : 7
                : 1603
                Affiliations
                [1] 1Department of Pathobiology, Functional Microbiology, Institute of Microbiology, University of Veterinary Medicine Vienna, Austria
                [2] 2Clinic for Ruminants, Department for Farm Animals and Herd Management, University of Veterinary Medicine Vienna, Austria
                [3] 3Department for Farm Animals and Herd Management, Institute of Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine Vienna, Austria
                [4] 4Chamber for Agriculture Vorarlberg Bregenz, Austria
                Author notes

                Edited by: Edward M. Fox, Commonwealth Scientific and Industrial Research Organisation (CSIRO), Australia

                Reviewed by: Antonio Valero, University of Córdoba, Spain; Karen Hunt, Teagasc–The Irish Agriculture and Food Development Authority, Ireland

                *Correspondence: Monika Ehling-Schulz monika.ehling-schulz@ 123456vetmeduni.ac.at

                This article was submitted to Food Microbiology, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2016.01603
                5061776
                27790200
                fc312666-55a4-4498-991b-44300c3b6af2
                Copyright © 2016 Kümmel, Stessl, Gonano, Walcher, Bereuter, Fricker, Grunert, Wagner and Ehling-Schulz.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 31 July 2016
                : 26 September 2016
                Page count
                Figures: 1, Tables: 2, Equations: 0, References: 49, Pages: 11, Words: 8425
                Funding
                Funded by: Sixth Framework Programme 10.13039/501100004965
                Award ID: 36272
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                staphylococcus aureus,dairy chain,mastitis,ftir spectroscopy,food safety,subtyping,metabolic fingerpriting

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