Genome Sequences of Lactococcus garvieae and Lactococcus petauri Strains Isolated from Traditional Montenegrin Brine Cheeses

Martinović, Aleksandra; Rosel, Adriana Cabal; Nisic, Andjela; Sucher, Jasmin; Stöger, Anna; Allerberger, Franz; Ruppitsch, Werner

doi:10.1128/mra.00546-21

Cited by 19 publications

(18 citation statements)

References 20 publications

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“…B1726 was sequenced using Illumina and ONT and assembled to a single circular contig. The genome has a size of 2,106,259 bp and a GC content of 38%, which is in line with previously sequenced L. garvieae and L. petauri genomes [14,68,69]. Initial whole genome sequence alignment proposed that the isolate belongs to the species L. garvieae as the closest relative was L. garvieae PAQ102015-99 with an average nucleotide identity (ANI) of 99.94% and a coverage of 97.39% (Additional file 1: Table S3).…”

Section: Genome Analysis and Identification Of The Putative Gar Opero...supporting

confidence: 86%

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Garvicin Q: characterization of biosynthesis and mode of action

et al. 2022

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Bacteriocins are ribosomally synthesized antimicrobial peptides, that either kill target bacteria or inhibit their growth. Bacteriocins are used in food preservation and are of increasing interest as potential alternatives to conventional antibiotics. In the present study, we show that Lactococcus petauri B1726, a strain isolated from fermented balsam pear, produces a heat-stable and protease-sensitive compound. Following genome sequencing, a gene cluster for production of a class IId bacteriocin was identified consisting of garQ (encoding for the bacteriocin garvicin Q), garI (for a putative immunity protein), garC, and garD (putative transporter proteins). Growth conditions were optimized for increased bacteriocin activity in supernatants of L. petauri B1726 and purification and mass spectrometry identified the compound as garvicin Q. Further experiments suggest that garvicin Q adsorbs to biomass of various susceptible and insusceptible bacteria and support the hypothesis that garvicin Q requires a mannose-family phosphotransferase system (PTSMan) as receptor to kill target bacteria by disruption of membrane integrity. Heterologous expression of a synthetic garQICD operon was established in Corynebacterium glutamicum demonstrating that genes garQICD are responsible for biosynthesis and secretion of garvicin Q. Moreover, production of garvicin Q by the recombinant C. glutamicum strain was improved by using a defined medium yet product levels were still considerably lower than with the natural L. petauri B1726 producer strain.Collectively, our data identifies the genetic basis for production of the bacteriocin garvicin Q by L. petauri B1726 and provides insights into the receptor and mode of action of garvicin Q. Moreover, we successfully performed first attempts towards biotechnological production of this interesting bacteriocin using natural and heterologous hosts.

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Section: Genome Analysis and Identification Of The Putative Gar Opero...supporting

confidence: 86%

“…Infections of humans by L. garvieae are often associated with contact to contaminated raw fish [9,10]. Interestingly, L. garvieae and L. petauri strains are widely distributed in the environment and were isolated not only from fish but also from various food products including raw milk, artisan cheese and vegetables [8,[11][12][13][14].…”

Section: Introductionmentioning

confidence: 99%

Garvicin Q: characterization of biosynthesis and mode of action

et al. 2022

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“…Clinical manifestation of infections caused by L. garvieae , L. petauri , or L. formosensis is similar and can include acute hemorrhagic septicemia, erratic swimming, lethargy, exophthalmia, anorexia, skin pigmentation changes, and moderate to high mortality in affected systems (Vendrell et al 2006; Shahin et al 2021). Additionally, commonly used diagnostic methods, such as biochemical identification kits, matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry, and published molecular methods, cannot discern among the three species or have not been fully validated (Martinovic et al 2021; Heckman et al 2022; Egger et al 2023). Currently, the most appropriate methods to differentiate among these three Lactococcus spp.…”

Section: Introductionmentioning

confidence: 99%

Detection and virulence of Lactococcus garvieae and L. petauri from four lakes in southern California

Abraham,

Yazdi,

Littman

et al. 2023

J Aqua Anim Hlth

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ObjectiveThe first objective of the study aimed to detect the presence of Lactococcus petauri, L. garvieae, and L. formosensis in fish (n = 359) and environmental (n = 161) samples from four lakes near an affected fish farm in California during an outbreak in 2020. The second objective was to compare the virulence of the Lactococcus spp. in Rainbow Trout Oncorhynchus mykiss and Largemouth Bass Micropterus salmoides.MethodsStandard bacterial culture methods were used to isolate Lactococcus spp. from brain and posterior kidney of sampled fish from the four lakes. Quantitative PCR (qPCR) was utilized to detect Lactococcus spp. DNA in fish tissues and environmental samples from the four lakes. Laboratory controlled challenges were conducted by injecting fish intracoelomically with representative isolates of L. petauri (n = 17), L. garvieae (n = 2), or L. formosensis (n = 4), and monitored for 14 days postchallenge (dpc).ResultLactococcus garvieae was isolated from the brains of two Largemouth Bass in one of the lakes. Lactococcus spp. were detected in 14 fish (8 Bluegills Lepomis macrochirus and 6 Largemouth Bass) from 3 out of the 4 lakes using a qPCR assay. Of the collected environmental samples, all 4 lakes tested positive for Lactococcus spp. in the soil samples, while 2 of the 4 lakes tested positive in the water samples through qPCR. Challenged Largemouth Bass did not show any signs of infection postinjection throughout the challenge period. Rainbow Trout infected with L. petauri showed clinical signs within 3 dpc and presented a significantly higher cumulative mortality (62.4%; p < 0.0001) at 14 dpc when compared to L. garvieae (0%) and L. formosensis (7.5%) treatments.ConclusionThe study suggests that qPCR can be used for environmental DNA monitoring of Lactococcus spp. and demonstrates virulence diversity between the etiological agents of piscine lactococcosis.

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“…Although the literature reveals that L. garvieae is a bacterial species with high genetic heterogeneity, fish isolates recovered from the same outbreak generally present a clonal structure. Considering that MLST is a well-established methodology for the study of L. garvieae genetic diversity (Ferrario et al, 2013; Lin et al, 2020; Reguera-Brito et al, 2016) and that L. petauri and L. garvieae share high genetic similarity (Goodman et al, 2017; Kotzamanidis et al, 2020; Martinovic et al, 2021; Ou et al, 2020), the MLST scheme for L. garvieae was used in this study to analyze L. petauri isolates. In addition, the MLST analysis allows the understanding of evolutionary stories between bacterial lineages and epidemiological tracking of bacteria (Maiden et al, 2013; Pérez-Losada et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Emerging fish pathogens Lactococcus petauri and L. garvieae in Nile tilapia (Oreochromis niloticus) farmed in Brazil

Egger¹,

Rosa²,

Pádua³

et al. 2022

Preprint

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Lactococcosis in fish has been associated with Lactococcus garvieae and the recently described L. petauri. However, the relevance of these emerging fish pathogens to Nile tilapia still requires thorough understanding. This study investigated lactococcosis outbreaks in Nile tilapia on Brazilian farms and characterized the isolates through molecular identification of the bacterial species, multilocus sequence typing (MLST) analysis, virulence to Nile tilapia, and antimicrobial susceptibility. Lactococcosis outbreaks were monitored from 2019 to 2022 throughout Brazil. The outbreaks occurred mainly during warmer months, and co-infections were observed in four farms, whereas concurrent bacterial infections were identified in all farms. Since the sequence of the 16S rRNA was not capable of differentiating between L. petauri and L. garvieae, Lactococcus spp. isolates were identified at the species level using the gyrB gene sequence. In total, 30 isolates were classified as L. petauri and two as L. garvieae. All L. petauri isolates were grouped in ST24, except for one isolate which belonged to the newly described ST47. A new ST was also described for the L. garvieae isolates identified, ST46. Furthermore, L. petauri ST24 and ST47 were characterized as singletons, whereas L. garvieae ST46 was grouped with ST16 and ST17 and formed CC17. For the challenge trial, an L. petauri ST24 isolate was chosen considering that this MLST lineage was the most frequently observed. L. petauri was reisolated from challenged Nile tilapia, confirming the pathogenicity of this bacterium to Nile tilapia. The infection in the fish progressed very rapidly, and within 48 h post-challenge clinical signs and the first mortalities were observed. The estimated LD50 was 5.74 x 103 CFU 15 days post-challenge. Provisional epidemiological cutoff values were determined for L. petauri for six antimicrobial agents from different drug classes. All isolates were characterized as wild type (WT) for neomycin and oxytetracycline, whereas 96.67 % of the isolates were characterized as WT for amoxicillin, erythromycin, and florfenicol, and 83.33 % were WT for norfloxacin. Of the 14 outbreaks analyzed, 12 were caused by L. petauri and two by L. garvieae. The gyrB gene sequence was used to differentiate L. petauri from L. garvieae and allowed for the correct identification of these pathogens. Two MSLT lineages of L. petauri were identified and ST24 was observed in different regions of the country, illustrating a rapid expansion of this bacterial lineage.

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Genome Sequences of Lactococcus garvieae and Lactococcus petauri Strains Isolated from Traditional Montenegrin Brine Cheeses

Cited by 19 publications

References 20 publications

Garvicin Q: characterization of biosynthesis and mode of action

Garvicin Q: characterization of biosynthesis and mode of action

Detection and virulence of Lactococcus garvieae and L. petauri from four lakes in southern California

Emerging fish pathogens Lactococcus petauri and L. garvieae in Nile tilapia (Oreochromis niloticus) farmed in Brazil

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