Screening for and characterization of Lactococcus lactis bacteriophages with high thermal resistance

Atamer, Zeynep; Dietrich, Jochen; Müller-Merbach, Mareile; Neve, Horst; Heller, Knut J.; Hinrichs, Jörg

doi:10.1016/j.idairyj.2008.10.012

Cited by 83 publications

(112 citation statements)

References 29 publications

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“…The dominance of 936 phages in the present metavirome study is consistent with most previous lactococcal phage isolation studies (6,18,36,(46)(47)(48)(49)(50)(51)(52)(53). Detection of mostly SCH and phage 7 and occasionally 645, ASCC406, 1727, and ⌽L.6 RBP variants may suggest the presence of a limited diversity of these phages in the dairies.…”

Section: Discussionsupporting

confidence: 92%

Metagenomic Analysis of Dairy Bacteriophages: Extraction Method and Pilot Study on Whey Samples Derived from Using Undefined and Defined Mesophilic Starter Cultures

Muhammed

Kot

Neve

et al. 2017

Appl Environ Microbiol

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Despite being potentially highly useful for characterizing the biodiversity of phages, metagenomic studies are currently not available for dairy bacteriophages, partly due to the lack of a standard procedure for phage extraction. We optimized an extraction method that allows the removal of the bulk protein from whey and milk samples with losses of less than 50% of spiked phages. The protocol was applied to extract phages from whey in order to test the notion that members of Lactococcus lactis 936 (now Sk1virus), P335, c2 (now C2virus) and Leuconostoc phage groups are the most frequently encountered in the dairy environment. The relative abundance and diversity of phages in eight and four whey mixtures from dairies using undefined mesophilic mixed-strain cultures containing Lactococcus lactis subsp. lactis biovar diacetylactis and Leuconostoc species (i.e., DL starter cultures) and defined cultures, respectively, were assessed. Results obtained from transmission electron microscopy and high-throughput sequence analyses revealed the dominance of Lc. lactis 936 phages (order Caudovirales, family Siphoviridae) in dairies using undefined DL starter cultures and Lc. lactis c2 phages (order Caudovirales, family Siphoviridae) in dairies using defined cultures. The 936 and Leuconostoc phages demonstrated limited diversity. Possible coinduction of temperate P335 prophages and satellite phages in one of the whey mixtures was also observed. IMPORTANCEThe method optimized in this study could provide an important basis for understanding the dynamics of the phage community (abundance, development, diversity, evolution, etc.) in dairies with different sizes, locations, and production strategies. It may also enable the discovery of previously unknown phages, which is crucial for the development of rapid molecular biology-based methods for phage burden surveillance systems. The dominance of only a few phage groups in the dairy environment signifies the depth of knowledge gained over the past decades, which served as the basis for designing current phage control strategies. The presence of a correlation between phages and the type of starter cultures being used in dairies might help to improve the selection and/or design of suitable, custom, and cost-efficient phage control strategies.KEYWORDS abundance, bacteriophages, Caudovirales, dairy, diversity, Lactococcus lactis, Leuconostoc, mesophilic, metavirome, Siphoviridae, starter culture, whey

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Section: Discussionsupporting

confidence: 92%

Metagenomic Analysis of Dairy Bacteriophages: Extraction Method and Pilot Study on Whey Samples Derived from Using Undefined and Defined Mesophilic Starter Cultures

Muhammed

Kot

Neve

et al. 2017

Appl Environ Microbiol

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“…These low to medium phage titers reflect the small number of Leuconostoc strains in undefined mixed-strain starter cultures (i.e., Ͻ10%) (6). Lactococcal phages, however, commonly accumulate in dairy samples at high numbers of approximately 10 9 PFU per ml of whey (25).…”

Section: Resultsmentioning

confidence: 99%

“…Negative staining of phage samples with 2% (wt/vol) uranyl acetate (Plano, Wetzlar, Germany) on freshly prepared carbon films (Plano) and transmission electron microscopy (TEM; Tecnai 10; FEI Company, Eindhoven, The Netherlands) at an acceleration voltage of 80 kV were performed as described earlier (25,26).…”

Section: Phages and Strainsmentioning

confidence: 99%

Classification of Lytic Bacteriophages Attacking Dairy Leuconostoc Starter Strains

Ali

Kot

Atamer

et al. 2013

Appl Environ Microbiol

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A set of 83 lytic dairy bacteriophages (phages) infecting flavor-producing mesophilic starter strains of the Leuconostoc genus was characterized, and the first in-depth taxonomic scheme was established for this phage group. Phages were obtained from different sources, i.e., from dairy samples originating from 11 German dairies (50 Leuconostoc pseudomesenteroides [Ln. pseudomesenteroides] phages, 4 Ln. mesenteroides phages) and from 3 external phage collections (17 Ln. pseudomesenteroides phages, 12 Ln. mesenteroides phages). All phages belonged to the Siphoviridae family of phages with isometric heads (diameter, 55 nm) and noncontractile tails (length, 140 nm). With the exception of one phage (i.e., phage ⌽LN25), all Ln. mesenteroides phages lysed the same host strains and revealed characteristic globular baseplate appendages. Phage ⌽LN25, with different Y-shaped appendages, had a unique host range. Apart from two phages (i.e., phages P792 and P793), all Ln. pseudomesenteroides phages shared the same host range and had plain baseplates without distinguishable appendages. They were further characterized by the presence or absence of a collar below the phage head or by unique tails with straight striations. Phages P792 and P793 with characteristic fluffy baseplate appendages could propagate only on other specific hosts. All Ln. mesenteroides and all Ln. pseudomesenteroides phages were members of two (host species-specific) distinct genotypes but shared a limited conserved DNA region specifying their structural genes. A PCR detection system was established and was shown to be reliable for the detection of all Leuconostoc phage types.

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“…In a recent study on phage dissemination in whey powders, it was shown that 94% of the samples contained lytic phages up to titres of 10 7 pfu g À1 (Wagner, 2012). In previous studies, heat inactivation has been studied in detail, and it was documented that most phages can withstand pasteurization processes of 72e75 C for 15e30 s (Atamer et al, 2009;Atamer & Hinrichs, 2010;Müller-Merbach, Neve, & Hinrichs, 2005;Quiberoni, Guglielmotti, & Reinheimer, 2003;Su arez & Reinheimer, 2002). Consequently, thermo-resistant phages are capable of propagating during cheese processing, resulting in phage titres up to 10 9 phages per mL in drained cheese whey (Neve, 1996;Neve, Berger, & Heller, 1995).…”

Section: Introductionmentioning

confidence: 99%

Application of a membrane technology to remove bacteriophages from whey

Samtlebe

Wagner

Neve

et al. 2015

International Dairy Journal

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Screening for and characterization of Lactococcus lactis bacteriophages with high thermal resistance

Cited by 83 publications

References 29 publications

Metagenomic Analysis of Dairy Bacteriophages: Extraction Method and Pilot Study on Whey Samples Derived from Using Undefined and Defined Mesophilic Starter Cultures

Metagenomic Analysis of Dairy Bacteriophages: Extraction Method and Pilot Study on Whey Samples Derived from Using Undefined and Defined Mesophilic Starter Cultures

Classification of Lytic Bacteriophages Attacking Dairy Leuconostoc Starter Strains

Application of a membrane technology to remove bacteriophages from whey

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