Antimicrobial activity, molecular typing and in vitro safety assessment of Lactococcus garvieae isolates from healthy cultured rainbow trout (Oncorhynchus mykiss, Walbaum) and rearing environment

Feito, Javier; Araújo, Carlos; Gómez‐Sala, Beatriz; Contente, Diogo; Campanero, Cristina; Arbulu, Sara; Saralegui, Claudia; Peña, Nuria; Muñoz-Atienza, Estefanía; Borrero, Juan; Campo, R. del; Hernández, Pablo E.; Cintas, Luis M.

doi:10.1016/j.lwt.2022.113496

Cited by 8 publications

(6 citation statements)

References 94 publications

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“…These results and observations are somewhat in agreement with previous studies. In this regard, ERIC-PCR was shown to be an easy and rapid strategy for the molecular fingerprinting of LAB, but, also, as observed by other authors, similarities observed in DNA fingerprinting techniques do not always directly correlate with similar phenotypic characteristics [42,63]. Furthermore, ERIC-PCR analyses suggested that the broad antimicrobial spectrum of Lc.…”

Section: St Parauberis Lmg225mentioning

confidence: 83%

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Antimicrobial Activity, Genetic Relatedness, and Safety Assessment of Potential Probiotic Lactic Acid Bacteria Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis) Used as Live Feed in Fish Larviculture

Contente,

Díaz-Formoso,

Feito

et al. 2024

Animals

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Aquaculture is a rapidly expanding agri-food industry that faces substantial economic losses due to infectious disease outbreaks, such as bacterial infections. These outbreaks cause disruptions and high mortalities at various stages of the rearing process, especially in the larval stages. Probiotic bacteria are emerging as promising and sustainable alternative or complementary strategies to vaccination and the use of antibiotics in aquaculture. In this study, potential probiotic candidates for larviculture were isolated from a rotifer-rearing tank used as the first live feed for turbot larvae. Two Lacticaseibacillus paracasei and two Lactiplantibacillus plantarum isolates were selected for further characterization due to their wide and strong antimicrobial activity against several ichthyopathogens, both Gram-positive and Gram-negative. An extensive in vitro safety assessment of these four isolates revealed the absence of harmful traits, such as acquired antimicrobial resistance and other virulence factors (i.e., hemolytic and gelatinase activities, bile salt deconjugation, and mucin degradation, as well as PCR detection of biogenic amine production). Moreover, Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR) analyses unveiled their genetic relatedness, revealing two divergent clusters within each species. To our knowledge, this work reports for the first time the isolation and characterization of Lactic Acid Bacteria (LAB) with potential use as probiotics in aquaculture from rotifer-rearing tanks, which have the potential to optimize turbot larviculture and to introduce novel microbial management approaches for a sustainable aquaculture.

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Section: St Parauberis Lmg225mentioning

confidence: 83%

“…Lactobacillus sp. 30A and Enterococcus faecium L50 were used as positive and negative controls for hdc and odc, and tdc, respectively [42].…”

Section: Biogenic Amine Production Pcr-detectionmentioning

confidence: 99%

Antimicrobial Activity, Genetic Relatedness, and Safety Assessment of Potential Probiotic Lactic Acid Bacteria Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis) Used as Live Feed in Fish Larviculture

Contente,

Díaz-Formoso,

Feito

et al. 2024

Animals

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“…Different approaches have been used to assess L. garvieae genetic diversity, including enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR), repetitive element palindromic-PCR (REP-PCR), and MLST (Feito et al, 2022; Ortega et al, 2020; Reguera-Brito et al, 2016; Torres-Corral and Santos, 2021). 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.…”

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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“…This is an emerging pathogen in aquaculture, causing lactococcosis, a disease that results in significant economic losses. Moreover, it could be considered a potential foodborne pathogen for humans [69]. This species, as lactic acid bacteria, is a component of native microbiota in raw milk and derived products [70], and also, it is associated with the fermentation of fish products [71], although the disease is linked with the consumption of contaminated raw fish [72,73].…”

Section: Bacterial Genus Presence In Water Samplesmentioning

confidence: 99%

“…In contrast, Lactococcus garvieae, which has been isolated in the flesh samples in the present study, is the etiological agent of lactococcosis, an ichthyopathology that affects rainbow trout farming. Thus, it would be important to prevent and control the presence of L. garvieae in fish farming [69,88].…”

Section: Bacterial Genus Presence In Rainbow Trout Samplesmentioning

confidence: 99%

Tracking Microbial Diversity and Hygienic-Sanitary Status during Processing of Farmed Rainbow Trout (Oncorhynchus mykiss)

Serrano Heredia,

Sánchez-Martín,

Romero Gil

et al. 2023

Foods

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Aquaculture is becoming a strategic sector for many national economies to supply the increasing demand for fish from consumers. Fish culture conditions and processing operations can lead to an increase in microbial contamination of farmed fish that may shorten the shelf-life of fish products and byproducts, and ready-to-eat fishery products. The objective of this study was to evaluate the hygienic-sanitary status of water, environment, and processing of fresh-farmed rainbow trout (Oncorhynchus mykiss) fillets produced in a local fish farm in Andalusia, Spain. To achieve this, a longitudinal study was carried out by collecting environmental (air and food-contact surfaces), water from fish ponds, and rainbow trout samples. Thereby, seven sampling visits were performed between February 2021 and July 2022, where foodborne pathogens and spoilage microorganisms, together with physicochemical parameters, were analysed in the collected samples. Further, microbial identification of microbiota was achieved through a culture-dependent technique using blast analysis of 16S RNA gene sequencing. The results showed that Listeria monocytogenes and Salmonella were not detected in the analysed samples. Regarding the hygienic-sanitary status of the fish farm, the slaughtering bath, the eviscerating machine and the outlet water from fish ponds presented the highest counts of coliforms, Enterobacteriaceae, and Aerobic Mesophilic Bacteria. Staphylococcus aureus and sulphite-reducing Clostridium were identified in the conveyor belts, fish flesh, and viscera. The 16S RNA identification confirmed the presence of viable spoilage bacteria such as Citrobacter gillenii, Macrococcus caseolyticus, Hafnia paralvei, Lactococcus lactis, Lactococcus cremoris, Klebsiella, Escherichia coli, Morganella morganii, and Shewanella. Three of these genera (Citrobacter, Hafnia, and Pseudomonas) were present in all types of samples analysed. The results evidenced potential transmission of microbial contamination from contaminated packaging belts and boxes, evisceration and filleting machines to flesh and viscera samples, thus the establishment of control measures should be implemented in fish farm facilities to extend the shelf-life of farmed fishery products.

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Antimicrobial activity, molecular typing and in vitro safety assessment of Lactococcus garvieae isolates from healthy cultured rainbow trout (Oncorhynchus mykiss, Walbaum) and rearing environment

Cited by 8 publications

References 94 publications

Antimicrobial Activity, Genetic Relatedness, and Safety Assessment of Potential Probiotic Lactic Acid Bacteria Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis) Used as Live Feed in Fish Larviculture

Antimicrobial Activity, Genetic Relatedness, and Safety Assessment of Potential Probiotic Lactic Acid Bacteria Isolated from a Rearing Tank of Rotifers (Brachionus plicatilis) Used as Live Feed in Fish Larviculture

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

Tracking Microbial Diversity and Hygienic-Sanitary Status during Processing of Farmed Rainbow Trout (Oncorhynchus mykiss)

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