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Giant freshwater prawn (Macrobrachium rosenbergii (MR)) is a significant aquafarm species commercially cultured in Taiwan. Intensive farming practices have led to the outbreak of Lactococcus garvieae (LG), which causes Lactococcosis in MR. Recently, LG has re‐emerged and the number of mortalities in prawn farms has increased in Taiwan. However, there is no preventative strategy described and a lack of knowledge on virulence factors and pathogenesis from LG in MR. The most virulent strain of L. garvieae from M. rosenbergii was screened in vivo among seven isolates selected for infectivity testing injecting 0.1 mL of 108 CFU/mL bacterial concentration. Among the seven isolates screened, L. garvieae 109‐6 resulted in 100% mortality within 3 days post‐infection. Furthermore, 109‐6 L. garvieae LD50 dosage from in MR was found to be 106 CFU/mL. Subsequently, the most virulent strain 109‐6 was sequenced using MinIon Nanopore sequencing. Results indicated that the LG genome yielded a protein‐coding of 3857 with 59 tRNA and 16 rRNA and no plasmid. Interestingly, the distribution of subsystems in the annotated genome revealed genes related to virulence, defence, and disease among LG 50 genes. Altogether, the virulent strain and its genome data revealed distinctive features of LG, which hinted toward its pathogenicity and could facilitate for better preventive strategies. [ABSTRACT FROM AUTHOR]

Lactococcosis is a common bacterial fish disease caused by Lactococcus garvieae, L. petauri and L. formosensis. Although there are different PCR‐based techniques to identify the etiological agent, none of these can differentiate these two bacteria without sequencing PCR‐amplified fragments. In the present study, we developed a multiplex PCR assay for simultaneous detection and differentiation of L. garvieae and L. petauri. The specificity of the primers was validated against the bacterial DNA of the targeted and non‐targeted bacteria. The sizes of the PCR amplicons were obtained as 204 bp for the DUF1430 domain‐containing protein gene of L. garvieae, 465 bp for the Lichenan permease IIC component gene of L. petauri, and 302 bp for the teichoic acid biosynthesis protein F gene of both L. garvieae and L. petauri. The PCR amplicons were clearly separated by agarose gel electrophoresis. The multiplex PCR assay did not produce any amplification products with the DNA of the non‐targeted bacteria. The multiplex PCR detection limits for L. garvieae and L. petauri were 5 and 4 CFU in pure culture and 50 and 40 CFU/g in spiked tissue samples, respectively. It takes less than 2 h from plate‐cultured bacteria and 3 h from tissue samples to get results. In conclusion, the developed multiplex PCR assay is a rapid, specific, accurate, and cost‐effective method for the detection and differentiation of L. garvieae and L. petauri and is suitable to be used for routine laboratory diagnosis of L. garvieae and L. petauri. [ABSTRACT FROM AUTHOR]

The aquaculture sector plays a vital role in global food security, yet it grapples with significant challenges posed by infectious diseases. Piscine lactococcosis is one of the significant threats in rainbow trout aquaculture due to its potential to cause severe economic losses through mortalities, reduced growth rates, and increased susceptibility to other pathogens. It poses challenges in disease management strategies, impacting the sustainability and profitability of rainbow trout farming. The current study focuses on the variations in serum blood parameters of farmed rainbow trout Oncorhynchus mykiss during a lactococcosis outbreak caused by Lactococcus garvieae. Blood samples were collected for biochemical analysis, fish were examined for parasites and bacteria, and DNA from bacterial colonies was PCR‐amplified and sequenced for identification. Overall, 13 biochemical parameters, including proteins, enzymes, lipids, chemicals, and minerals, were measured in serum blood samples from both diseased and healthy fish. The results indicate significant alterations in the levels of these parameters during the outbreak, highlighting the impact of infections on the blood profile of farmed rainbow trout. Urea levels were significantly higher in diseased fish compared to controls, and creatinine, phosphorus, and magnesium also showed similar trends. Alanine aminotransferase and total protein levels were higher in control fish. Chloride levels differed significantly between groups. Iron levels were higher in controls and lower in diseased fish. No significant differences were found in other parameters. This study reveals significant changes in serum blood parameters of rainbow trout during a lactococcosis outbreak caused by L. garvieae. These changes highlight the potential of these parameters as tools for monitoring health status, stress, and aquaculture management. Continuous monitoring can provide valuable insights into disease severity and overall fish health, aiding in the development of improved management practices. The presented data contribute to understanding the pathophysiology of piscine lactococcosis and developing effective mitigation strategies for farmed rainbow trout. [ABSTRACT FROM AUTHOR]

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ABSTRACT The gut microbiota, a pivotal component of the intestinal mucosal barrier, is critical for host resistance to enteric pathogen infection. Here, we report a novel function of the potentially probiotic Lactococcus garvieae strain LG1 (L. garvieae strain LG1) in maintaining intestinal mucosal barrier integrity and protecting against foodborne Clostridium perfringens (C. perfringens) infection. L. garvieae was isolated from the intestinal contents of Chinese Mongolian sheep (MS) and exhibited potential probiotic properties. In a C. perfringens enterocolitis model, L. garvieae-pretreated mice were less susceptible to C. perfringens infection compared with Phosphate buffered solution (PBS)-pretreated mice, which manifested as higher survival rates, lower pathogen loads, less weight loss, mild clinical symptoms and intestinal damage, and minor inflammation. Further mechanistic analysis showed that L. garvieae could ameliorate the disruption of intestinal permeability and maintain the integrity of the intestinal mucosal barrier by promoting the expression of tight junction proteins and mucoproteins. Moreover, L. garvieae was also able to facilitate antimicrobial peptide expression and ameliorate dysbiosis of the gut microbiota caused by C. perfringens. Together, these findings highlight the prospect of immunomodulatory potentially probiotic L. garvieae and might offer valuable strategies for prophylaxis and/or treatment of pathogenic C. perfringens mucosal infection.IMPORTANCEC. perfringens necrotic enteritis leads to losses of about US $2 billion to the poultry industry worldwide every year. Worse, US Centers for Disease Control and Prevention (CDC) has estimated that C. perfringens causes nearly 1 million foodborne illnesses in the United States annually. Nowadays, the treatment recommendation is a combination of a broad-spectrum synergistic penicillin with clindamycin or a carbapenem, despite growing scientific concern over antibiotic resistance. The global understanding of the gut microbiome for C. perfringens infection may provide important insights into the intervention. L. garvieae originated from Mongolian sheep intestine, exhibited potentially probiotic properties, and was able to limit C. perfringens enterocolitis and pathogenic colonization. Importantly, we found that L. garvieae limits C. perfringens invasion via improving intestinal mucosal barrier function. Also, L. garvieae alleviates C. perfringens-induced gut microbiota dysbiosis. It allowed us to convince that utilization of probiotics to promote protective immunity against pathogens infection is of pivotal importance.

Lactococcus garvieae is a known fish pathogen associated with numerous aquacultural outbreaks. In humans, L. garvieae primarily causes infective endocarditis, but infections involving other organs have also been reported. We report the first case of ruptured infectious intracranial aneurysm associated with L. garvieae bacteraemia without concomitant infective endocarditis. The diagnosis of a left distal posterior cerebral artery mycotic aneurysm was based on a computed tomography angiogram, catheter angiogram and histopathological examination of the resected aneurysm. Here, we review the literature on human L. garvieae infections and describe the clinical characteristics, risk factors, management and outcomes of the cases identified to date.

In this study, a strain, named S5-4, with high esterase activity was obtained from Nongxiangxing Daqu by primary and secondary screening, and subjected to whole genome sequencing, esterase annotation and metabolic pathway analysis. The strain exhibited a maximal ratio of transparent zone diameter to colony diameter (D/d) of 2.27 ± 0.02, and it was identified as Lactococcus garvieae. Its morphological, physiological, and biochemical characteristics were determined, and its esterase activity was measured to be 15.74 U/mL. This strain produced (0.345 0 ± 0.160 0) g/L of ethyl acetate and (0.298 3 ± 0.230 0) g/L of ethyl lactate after anaerobic fermentation at 35 ℃ for 15 days. The genome size of S5-4 was determined to be 2 191 113 bp in length, with a GC content of 37.99%. A total of 1 486 genes, including 4 rRNA genes, 69 tRNA genes, and 34 ncRNA genes were predicted. In total, 11 esterase genes were annotated in the genome of S5-4 including phosphatases (five intracellular ones and two membrane ones), acyl-CoA dehydrogenase (intracellular), triacylglycerol lipase (extracellular), carboxylase (extracellular), and hemicellulose esterase (intracellular). Based on functional genomic annotation, the metabolic pathways to produce ethyl acetate and ethyl lactate from starch and galactose as carbon sources were constructed and found to be consistent with the experimental results. Given its ability to produce esterase, the strain could be used as a biocatalyst for esterification in baijiu fermentation or as a flavor enhancer in Daqu production.

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Lactococcus garvieae (L. garvieae) is a pathogenic bacterium that is Gram-positive and catalase-negative (GPCN), and it is capable of growing in a wide range of environmental conditions. This bacterium is associated with significant mortality and losses in fisheries, and there are concerns regarding its potential as a zoonotic pathogen, given its presence in cattle and dairy products. While we have identified and characterized virulent strains of L. garvieae through phenotyping and molecular typing studies, their impact on mammary tissue remains unknown. This study aims to investigate the pathogenicity of strong and weak virulent strains of L. garvieae using in vivo mouse models. We aim to establish MAC-T cell model to examine potential injury caused by the strong virulent strain LG41 through the TLR2/NLRP3/NF-kB pathway. Furthermore, we assess the involvement of NLRP3 inflammasome-mediated pyroptosis in dairy mastitis by silencing NLRP3. The outcomes of this study will yield crucial theoretical insights into the potential mechanisms involved in mastitis in cows caused by the L. garvieae-induced inflammatory response in MAC-T cells. [ABSTRACT FROM AUTHOR]

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Background and Aim: The rapid development of aquaculture as a major food sector is accompanied by challenges, including diseases that affect tilapia farming worldwide. One such infectious disease caused by Streptococcus agalactiae poses a serious threat to tilapia populations. Probiotics have emerged as a potentially safe preventive measure against S. agalactiae infection. However, antimicrobial resistance from antibiotic-resistant bacteria remains a concern because it can lead to the spread of resistant bacteria and serve as a reservoir of antibiotic-resistant genes in fishes and the surrounding environment. This study aimed to identify candidate probiotic bacteria capable of promoting tilapia growth, providing resistance to S. agalactiae infection, devoid of potential pathogenicity, and free from antibiotic resistance genes. Subsequently, the performance of these probiotic candidates in tilapia was evaluated. Materials and Methods: Lactococcus garvieae, Priestia megaterium, Bacterium spp., Bacillus megaterium, Bacillus subtilis, and Bacillus pumilus were examined to assess their antibacterial properties, hemolytic patterns, and antibiotic resistance genes. We used the specific primers tetA, tetB, tetD, tetE, tetO, tetQ, ermB, and qnrS that were used for antibiotic resistance gene detection. In vivo probiotic efficacy was evaluated by administering probiotic candidates in tilapia feed at a concentration of 1 × 106 colonies/mL/50 g of feed over a 60-day maintenance period. Resistance to S. agalactiae infection was observed for 14 days after the challenge test. Results: Lactococcus garvieae, P. megaterium, and Bacterium spp. were identified as promising probiotic candidates among the bacterial isolates. On the other hand, B. megaterium, B. subtilis, and B. pumilus carried resistance genes and exhibited a β hemolytic pattern, rendering them unsuitable as probiotic candidates. The selected probiotic candidates (L. garvieae, P. megaterium, and Bacterium spp.) demonstrated the potential to enhance tilapia growth, exhibited no pathogenic tendencies, and were free from antibiotic resistance genes. Supplementation with L. garvieae and Bacterium spp. enhanced tilapia resistance to S. agalactiae infection, whereas P. megaterium supplementation showed an insignificant survival rate compared with controls after the challenge test period. Conclusion: Probiotics, particularly L. garvieae, P. megaterium, and Bacterium spp., enhance growth and resistance against S. agalactiae infection, without harboring antibiotic resistance genes. Selecting probiotic candidates based on antibiotic resistance genes is essential to ensure the safety of fish, the environment, and human health.

In this study, we investigated the dietary effects of dandelion root extract (DRE) on growth performance, body composition, serum biochemical parameters, intestinal and liver histology, and fish resistance against Lactococcus garvieae infection in the rainbow trout. In total, 540 rainbow trout fry (22.05±1.740 g and 13.03±0.470 cm) were used. Six experimental groups (control, 2.5 mL/kg (DRE-1), 5 mL/kg (DRE-2), 10 mL/kg (DRE-3), 20 mL/kg (DRE-4), and 40 mL/kg (DRE-5) dandelion root extract) with three replicates were formed. In each tank, 30 fish were placed and fed for 90 days, and the volume of water was adjusted to 80 L with a flow-through system. The results showed that the final fish weight, weight gain, specific growth rate, protein efficiency ratio, and protein deposition rate were significantly higher in the DRE-4 group, while the feed conversion ratio and fat deposition ratio were significantly lower compared to that in the control group (P<0.05). The viscerosomatic index and the hepatosomatic index decreased in the experimental groups depending on the concentration of the extract used, while the condition factor was higher than that in the control (P<0.05). The DRE significantly decreased glucose, cholesterol, triglyceride, low-density lipoprotein, alanine aminotransferase, and aspartate aminotransferase levels in blood serum in the fish from the experimental groups, but it significantly increased high-density lipoprotein, alkaline phosphatase, and total protein levels (P<0.05). Similarly, the DRE positively affected intestinal histology by increasing the stratum compactum, submucosa, lamina propria, villi length, villi width, absorption area, tunica muscularis thickness, mucosal folds, and goblet cell count. In the liver, depending on the high-fat diet (crude lipid = 20%), vacuolization areas, hepatocellular degeneration areas, apoptotic hepatocyte nucleus, and necrotic areas recorded in the control group decreased or even disappeared completely in the extract groups. The results of the challenge test showed that adding 2.5 mL/kg of the DRE to the feed positively affected the disease resistance against Lactococcus garvieae infection. The results of the quadratic polynomial regression showed that an average level of 23.91±1.048 mL/kg of DRE would be suitable for optimum fish growth. [ABSTRACT FROM AUTHOR]

Background and Aim: The rapid development of aquaculture as a major food sector is accompanied by challenges, including diseases that affect tilapia farming worldwide. One such infectious disease caused by Streptococcus agalactiae poses a serious threat to tilapia populations. Probiotics have emerged as a potentially safe preventive measure against S. agalactiae infection. However, antimicrobial resistance from antibiotic-resistant bacteria remains a concern because it can lead to the spread of resistant bacteria and serve as a reservoir of antibiotic-resistant genes in fishes and the surrounding environment. This study aimed to identify candidate probiotic bacteria capable of promoting tilapia growth, providing resistance to S. agalactiae infection, devoid of potential pathogenicity, and free from antibiotic resistance genes. Subsequently, the performance of these probiotic candidates in tilapia was evaluated. Materials and Methods: Lactococcus garvieae, Priestia megaterium, Bacterium spp., Bacillus megaterium, Bacillus subtilis, and Bacillus pumilus were examined to assess their antibacterial properties, hemolytic patterns, and antibiotic resistance genes. We used the specific primers tetA, tetB, tetD, tetE, tetO, tetQ, ermB, and qnrS that were used for antibiotic resistance gene detection. In vivo probiotic efficacy was evaluated by administering probiotic candidates in tilapia feed at a concentration of 1 x 106 colonies/mL/50 g of feed over a 60-day maintenance period. Resistance to S. agalactiae infection was observed for 14 days after the challenge test. Results: Lactococcus garvieae, P. megaterium, and Bacterium spp. were identified as promising probiotic candidates among the bacterial isolates. On the other hand, B. megaterium, B. subtilis, and B. pumilus carried resistance genes and exhibited a β hemolytic pattern, rendering them unsuitable as probiotic candidates. The selected probiotic candidates (L. garvieae, P. megaterium, and Bacterium spp.) demonstrated the potential to enhance tilapia growth, exhibited no pathogenic tendencies, and were free from antibiotic resistance genes. Supplementation with L. garvieae and Bacterium spp. enhanced tilapia resistance to S. agalactiae infection, whereas P. megaterium supplementation showed an insignificant survival rate compared with controls after the challenge test period. Conclusion: Probiotics, particularly L. garvieae, P. megaterium, and Bacterium spp., enhance growth and resistance against S. agalactiae infection, without harboring antibiotic resistance genes. Selecting probiotic candidates based on antibiotic resistance genes is essential to ensure the safety of fish, the environment, and human health. [ABSTRACT FROM AUTHOR]

This mini review deals with some controversial non-starter lactic acid bacteria (NSLAB) species known to be both human and animal pathogens but also health-promoting and probiotic. The focus is on Lactococcus garvieae, two Streptococcus species (S. uberis and S. parauberis), four Weissella species (W. hellenica, W. confusa, W. paramesenteroides, and W. cibaria), and Mammalicoccus sciuri, which worldwide, are often found within the microbiotas of different kinds of cheese, mainly traditional artisanal cheeses made from raw milk and/or relying on environmental bacteria for their ripening. Based on literature data, the virulence and health-promoting effects of these bacteria are examined, and some of the mechanisms of these actions are reviewed. Additionally, their possible roles in cheese ripening are also discussed. The analysis of the literature data available so far showed that, in general, the pathogenic and the beneficial strains, despite belonging to the same species, show somewhat different genetic constitutions. Yet, when the safety of a given strain is assessed, genomic analysis on its own is not enough, and a polyphasic approach including additional physiological and functional tests is needed. [ABSTRACT FROM AUTHOR]

Lactococcosis, caused by Lactococcus garvieae, is an acute hemorrhagic septicemia in fish recorded in marine and freshwater aquaculture during the summer months. In 2020–2021, several sea cage Pompano farms recorded sudden fish mortality events. Based on the results of phenotypic and biochemical tests, L. garvieae was predicted to be the cause. PCR with L. garvieae specific primers (pLG1 and pLG2) targeting the 16S rRNA region further confirmed the etiological agent as L. garvieae after amplifying an 1100 base pairs (bp) product. Furthermore, the 16S rRNA sequences of the two representative strains (AOD109‐196‐2B and AOD110‐215‐2B) shared 99.81% identity with L. garvieae (GenBank accession number: MT597707.1). The genetic profiles of the strains were classified using pulsed‐field gel electrophoresis after digestion with SmaI and ApaI, which clustered our strains under the same pulsotype. Multiplex PCR targeting the capsule gene cluster and serotype‐specific PCR collectively showed that the strains were non‐capsulated; thus, they belonged to serotype I. An experimental infection was designed to fulfil Koch's postulates by infecting healthy Pompano with case‐driven L. garvieae strains (AOD109‐196‐2B and AOD110‐215‐2B) with a cumulative mortality of 70%. Overall, L. garvieae infection in Pompano emphasizes the need for better monitoring and control procedures in aquaculture settings. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae is a Gram-positive coccus that can be easily misidentified as Enterococcus spp. or streptococci. Infection with L. garvieae is associated with the consumption of raw fish and unpasteurized dairy products. Although rare, it can cause infective endocarditis (IE). Herein, we report a case in which aortic valve replacement (AVR) was required for IE caused by L. garvieae.A 79-year-old Japanese man with a history of hypertension, myocardial infarction, gastroesophageal reflux disease (GERD), and abdominal aortic aneurysm presented with loss of appetite, myalgia, and difficulty in moving. Physical examination revealed a diastolic murmur, an Osler’s node on the right first toe, dental caries, and a palpable spleen, suggesting IE. Transthoracic echocardiography revealed a large, mobile vegetation on the aortic valve, which was associated with severe aortic regurgitation. Blood cultures revealed L. garvieae. The patient received antibiotic therapy, underwent AVR, and recovered without major complications. To date, 30 cases of L. garvieae-associated IE have been reported. We reviewed and summarized all cases of L. garvieae-associated IE including our case.

Dietary supplementation of immunostimulants might be effective to reduce the economic losses due to infectious diseases and the use of antibiotics in aquaculture. To investigate the immune response of interleukin-12 (IL-12) in yellowtail Seriola quinqueradiata to heat-killed Lactobacillus plantarum strain L-137 (HK L-137), we performed a leukocyte culture, feeding trial with diets containing L-137 and an immersion challenge with Lactococcus garvieae. IL-12 (IL-12p70) is a heterodimeric cytokine composed of IL-12p35 and IL-12p40 subunits. In the yellowtail-leukocyte culture, HK L-137 treatment stimulated the mRNA expression of one IL12p35 subunit (p35a) and all IL12p40 subunits (p40a, p40b, and p40c) in a dose-dependent manner. Furthermore, mRNA expression of type-I helper (Th-1) cytokine (tumor necrosis factor α, TNF-α, and interferon γ, IFN-γ) was also stimulated by HK L-137. After 6 weeks of feeding yellowtails with diets containing 0, 20, and 100 ppm of HK L-137, the mRNA expression of p35a and p40b in the spleen leukocytes increased with the dietary concentration of HK L-137, and that of p40b, p40c, and ifng in the head kidney leukocytes were the highest in the 20 ppm HK L-137 group. Survival rates in the 20 ppm HK L-137 group after immersion challenge with L. garvieae were significantly higher than the control (0 ppm of HK L-137). The 100 ppm HK L-137 group did not significantly suppress mortality. HK L-137 showed immunostimulant activity by increasing the expression of il-12, tnfa, and ifng mRNA in both in vitro and in vivo tests in yellowtail. Our results suggest that dietary supplementation with 20 ppm HK L-137 is the most efficient dose for improving immunity in yellowtail. Furthermore, a high dose of HK L-137 and/or long-term feeding of a diet containing HK L-137 might suppress the immune response, which probably decreases the survival rate of fish. To maintain a high immune response in yellowtail, the optimal dietary concentration of HK L-137 and/or feeding regime should be investigated further.

Lactococcus petauri is a recently described species of the genus Lactococcus. It was reported as an etiological agent of piscine lactococcosis together with Lactococcus garvieae. L. garvieae was already described as an opportunistic pathogen in human infections, with a potential zoonotic role. This paper represents the first report of a human urinary tract infection caused by L. petauri. A 91-year-old man was admitted to the emergency department for a femur fracture consequent to a domestic accident. The fracture was reduced by surgery and a catheterized specimen urine culture revealed a high bacterial load sustained by Gram-positive cocci, identified by Vitek 2 compact as L. garvieae, and subsequently as L. petauri through Internal Transcribed spacer 16S-23S r-RNA amplification. The number of L. petauri infections in humans is expected to rise in the near future mainly due to diagnostic improvement. A dedicated survey on L. garvieae and L. petauri infections in humans should be performed to better understand their role as pathogens and as zoonotic agents. [ABSTRACT FROM AUTHOR]

The isolation and characterization of bacterial species Lactococcus garvieae, previously unreported in whiteleg shrimp, Penaeus vannamei, has now been identified in the species. The pathogen was recovered from an affected shrimp farm in southern Taiwan. Bacterial characterization first identified the isolate as Gram‐positive cocci, and biochemical profiles demonstrated that the causative agent of mortality was 97% L. garvieae. The bacterial cell DNA resulted in amplification of 1522 bp with 99.6% confirmation by PCR analysis. The phylogenetic tree revealed 100% evolutionary similarity among previously isolated strains. Experimental infection further confirmed higher susceptibility of whiteleg shrimp to L. garvieae in waters of lower salinity, particularly 5 ppt, than in higher salinity. Histopathological analysis showed severely damaged hepatopancreas with necrotized, elongated, collapsed tubules, dislodged membranes and granuloma formation in infected shrimp. Transmission electron microscopy observation indicated a hyaluronic acid capsular layer surrounding bacterial cell which is a virulence factor of L. garvieae and likely responsible for immunosuppression and higher mortality of shrimp cultured in lower salinity. Collectively, these findings report the first isolation of L. garvieae from whiteleg shrimp and shed new light on the disease that threatens the highly valuable species and accentuates the need for finding a solution. [ABSTRACT FROM AUTHOR]

The emergence of antibiotic‐resistant pathogenic strains of Lactococcus garvieae serotype II isolated from fish in Japan has become a growing concern in recent years. The data on drug susceptibility and its associated resistance mechanism are limited. Therefore, the present study was conducted to determine the minimum inhibitory concentrations (MICs) of chemotherapeutic agents against 98 pathogenic strains of emerging Lactococcus garvieae serotype II isolated from fish from six different prefectures in Japan from 2018 to 2021. The tested strains were resistant to erythromycin, lincomycin and tiamulin. PCR amplification revealed the presence of erm(B) in all erythromycin‐resistant strains, while a conjugation experiment confirmed that these strains carried erm(B) that could be transferred to recipient Enterococcus faecalis OG1RF with frequencies from 10−4 to 10−6 per donor cells. Nucleotide sequencing of the representative isolated plasmid pkh2101 from an erythromycin‐resistant strain showed that it was a 26,850 bp molecule with an average GC content of 33.49%, comprising 31 CDSs, 13 of which remained without any functional annotation. Comparative genomic analysis suggested that pkh2101 shared the highest similarity (97.57% identity) with the plasmid pAMbeta1, which was previously isolated clinically from Enterococcus faecalis DS‐5. This study provides potential evidence that the plasmid harbouring erm(B) could be a source of antibiotic resistance transmission in emerging L. garvieae infection in aquaculture. [ABSTRACT FROM AUTHOR]

Lactococcosis, caused by members of the genus Lactococcus, represents a devastating disease inducing mass mortalities and economic losses in many fish species worldwide. The present work aimed to compare the whole genome sequences of three different serotypes of Lactococcus garvieae isolated from diseased cultured striped jack (Pseudocaranx dentex) in Ehime prefecture, Japan. The three serotypes showed different virulence in the challenge test using Japanese amberjack (Seriola quinqueradiata). The genome sequencing revealed that two of the strains (serotype I and serotype III) were identified as L. garvieae, while the third strain (serotype II) was identified as L. formosensis. The chromosome sizes of the three serotypes ranged from 1.9 to 2.0 Mb; the GC content ranges were 38.2 to 38.9%; and the numbers of predicted protein‐coding sequences (CDSs) were from 1922 to 1959. Only the serotype II harbours two plasmids, sizes of around 14 kb and 9 kb. The detected virulence factors varied among the different serotypes with some shared factors like adherence, anti‐phagocytosis, secretion system, toxin (haemolysin), serum resistance, antimicrobial resistance and others. The genomes also contained factors responsible for resistance to toxic compounds. The genome of the serotype III tended to encode more prophage regions than the other serotypes. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae is one of the primary pathogens causing the severe disease to golden pompano (Trachinotus ovatus). Vaccination is the most effective measure of preventing the occurrence and spread of infectious diseases. However, no commercially available vaccine protects T. ovatus against L. garvieae infection. In the present study, the formalin-inactivated L. garvieae vaccines, consisting of bacterin alone (aqueous vaccine) and combined with oil adjuvant Montanide™ ISA 763 A VG (vaccine+763A), were prepared and administered to T. ovatus by intraperitoneal injection. The protective efficacy was evaluated by testing the relative percent survival (RPS), serum enzyme activity, and expression levels of immune-related genes in the tissues of T. ovatus after immunization. The results indicated that the relative percentage survival (RPS) values of aqueous vaccine and vaccine+763A were 80% and 90.8% at 8 weeks post-vaccination. The level of antibody titer, lysozyme activities, superoxide dismutase, and catalase in the two vaccinated fish groups increased significantly compared to that in the control group. Moreover, the expression levels of immune-related genes IgM, TNF-a, IL-8, MHCI-a, MHCII, CD4, CD8a, and IL-1ß in all the tested tissues from T. ovatus were significantly enhanced after inoculation with aqueous vaccine and vaccine+763A. These findings demonstrated that the developed vaccine combined with oil adjuvant ISA 763 A VG improved immune responses and provided good protective efficacy against L. garvieae infection in the golden pompano culture. [ABSTRACT FROM AUTHOR]

Accurate and rapid determination of bacterial disease agents of fish is an important step for sustainable and efficient aquaculture production. In general, biochemical and molecular methods are used for pathogen detection but they are usually time-consuming and required qualified personnel. Recently spectroscopic methods are preferred in clinical and food microbiology and declared as a promising alternative method for pathogens diagnosis with many advantages. In this study, the significant spectra of three important bacterial fish pathogens (Lactococcus garvieae, Vibrio anguillarum and Yersinia ruckeri) were determined by Raman spectroscopy. The first data of the pathogens were obtained and the distinctive differences in polysaccharides, nucleic acids, fatty acids and amino acids were identified. This preliminary study aimed to be pioneer for further studies in aquaculture and veterinary microbiology toward developing an alternative method for routine identification. [ABSTRACT FROM AUTHOR]

Lactococci are Gram-positive cocci that occur in short chains or pairs and are traditionally considered to be of low virulence in human. Most species are not associated with human disease. There are few reports regarding Lactococcus isolation in humans and the clinical significance of this rarely-encountered genus is unknown. Here, we report a case of infectious spondylitis due to Lactococcus garvieae confirmed by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry (MALDI-TOFMS). A 77-year-old man was admitted to our hospital with back pain that had lasted 5 days. He had diabetes mellitus, hypertension, and histories of pulmonary tuberculosis and endovascular aneurysm repair due to an abdominal aortic aneurysm. Magnetic resonance imaging of his spine revealed paravertebral enhancement on T8-9 and a compression fracture on the lower endplate of T8. On blood cultures, L. garvieae was identified by MALDI-TOF MS. To our knowledge, this is the first report of spondylitis caused by L. garvieae in Korea. In this context, we reviewed non-endocarditis cases due to L. garvieae reported in the English-language literature to summarize its clinical features and outcomes. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically significant infectious disease affecting farmed rainbow trout. L. garvieae had been considered the only cause of lactococcosis for a long time; however, L. petauri, another species of the genus Lactococcus, has lately been linked to the same disease. The genomes and biochemical profiles of L. petauri and L. garvieae have a high degree of similarity. Traditional diagnostic tests currently available cannot distinguish between these two species. The aim of this study was to use the transcribed spacer (ITS) region between 16S rRNA and 23S rRNA as a potential useful molecular target to differentiate L. garvieae from L. petauri, saving time and money compared to genomics methods currently used as diagnostic tools for accurate discrimination between these two species. The ITS region of 82 strains was amplified and sequenced. The amplified fragments varied in size from 500 to 550 bp. Based on the sequence, seven SNPs were identified that separate L. garvieae from L. petauri. The 16S-23S rRNA ITS region has enough resolution to distinguish between closely related L. garvieae and L. petauri and it can be used as a diagnostic marker to quickly identify the pathogens in a lactococcosis outbreak. [ABSTRACT FROM AUTHOR]

Catfish species Pangasius nasutus, or locally known as Patin Buah is one of a popular aquaculture product in Malaysia. Probiotic strain is an attractive alternative to conventional antibiotics in managing aquaculture diseases. Pangasius can be a source of bacterial strain in developing suitable probiotic useful in Pangasius rearing. This study highlights on the isolation of Lactic Acid Bacteria (LAB) strains from the selected tissues from P. nasutus which are the heart, stomach, and intestines. These strains were subjected to morphological, biochemical, and genotypic characterisations. Three different strains were isolated from fish tissues, H-Hn from heart; S-Hn from stomach; and I-Sk from intestine. Biochemical characterisation were consistent with ribosomal rRNA sequencing, in which strain H-Hn (acc. No. MW504962) is highly similar (100%) to Lactococcus lactis, and both I-Sk (MW504964) and S-Hn (MW504963) are similar (99%) to Lactococcus garvieae. All strains appeared to be non-spore forming, non-motile, Gram-positive coccus, catalase and citrate negative, and fermenting sugar. Unlike H-Hn, both H-Sk and I-Sk strains were able to grow at 6.5% NaCl and at higher temperature (45°C). The antimicrobial properties were assessed using agar disk diffusion assay against several indicator organisms (Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Staphylococcus aureus and Vibrio parahaemolyticus). These strains exhibited varying degree of inhibitory properties against the indicator organisms. Strain H-Hn inhibited E. coli and V. parahaemolyticus. Meanwhile, strains I-Sk and S-Hn showed a rather broad-spectrum inhibition. From P. nasutus, other L. lactis strain, a pathogenic species from L garvieae could also be found especially in their gut tissues. The presence of L. garvieae in commercial P. nasutus should raise some concern to those who like to consume this fish. Meanwhile, the LAB strains isolated has probiotics potential that can be commercially used in the managing fish diseases in aquaculture. [ABSTRACT FROM AUTHOR]

Owing to an increasing number of infections in adults, Lactococcus (L.) garvieae has gained recognition as an emerging human pathogen, causing bacteraemia and septicaemia. In September 2020, four paediatric onco-hematologic patients received a platelet concentrate from the same adult donor at Bambino Gesù Children’s Hospital IRCCS, Rome. Three of four patients experienced L. garvieae sepsis one day after transfusion. The L. garvieae pediatric isolates and the donor’s platelet concentrates were retrospectively collected for whole-genome sequencing and shot-gun metagenomics, respectively (Illumina HiSeq). By de novo assembly of the L. garvieae genomes, we found that all three pediatric isolates shared a 99.9% identity and were characterized by 440 common SNPs. Plasmid pUC11C (conferring virulence properties) and the temperate prophage Plg-Tb25 were detected in all three strains. Core SNP genome-based maximum likelihood and Bayesian trees confirmed their phylogenetic common origin and revealed their relationship with L. garvieae strains affecting cows and humans (bootstrap values >100 and posterior probabilities = 1.00). Bacterial reads obtained by the donor’s platelet concentrate have been profiled with MetaPhlAn2 (v.2.7.5); among these, 29.9% belonged to Firmicutes, and 5.16% to Streptococcaceae (>97% identity with L. garvieae), confirming the presence of L. garvieae in the platelet concentrate transfusion. These data showed three episodes of sepsis for the first time due to a transfusion-associated transmission of L. garvieae in three pediatric hospitalized hematology patients. This highlights the importance to implement the screening of platelet components with new human-defined pathogens for ensuring the safety of blood supply, and more broadly, for the surveillance of emerging pathogens.

Lactococcus garvieae is a Gram-positive cocci that has been known to be a fish pathogen, and considered as a low virulence organism rarely associated with human infection. We report a case of acute cholangitis with common bile duct (CBD) stone and bacteremia by L. garvieae bacteremia in a 70-year-old male. The patient presented with epigastric pain and was diagnosed with two CBD stones. Blood culture obtained prior to empiric antimicrobial therapy with ceftizoxime sodium showed growth with Escherichia coli and L. garvieae. The bacteria were confirmed by matrix-assisted desorption/ionization time-of-flight mass spectrometry. Initial attempt at endoscopic biliary drainage failed, and the patient underwent percutaneous transhepatic biliary drainage and subsequent stone removal. He occasionally ingested raw fish and had a history of gastric ulcer with acid suppression therapy, which could be possible risk factors for L. garvieae infection. This is the first case of L. garvieae bacteremia in acute cholangitis. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae is a main ichthyopathogen in rainbow trout (Oncorhynchus mykiss, Walbaum) farming, although bacteriocinogenic L. garvieae with antimicrobial activity against virulent strains of this species have also been identified. Some of the bacteriocins characterized, such as garvicin A (GarA) and garvicin Q (GarQ), may show potential for the control of the virulent L. garvieae in food, feed and other biotechnological applications. In this study, we report on the design of Lactococcus lactis strains that produce the bacteriocins GarA and/or GarQ, either alone or together with nisin A (NisA) or nisin Z (NisZ). Synthetic genes encoding the signal peptide of the lactococcal protein Usp45 (SPusp45), fused to mature GarA (lgnA) and/or mature GarQ (garQ) and their associated immunity genes (lgnI and garI, respectively), were cloned into the protein expression vectors pMG36c, which contains the P32 constitutive promoter, and pNZ8048c, which contains the inducible PnisA promoter. The transformation of recombinant vectors into lactococcal cells allowed for the production of GarA and/or GarQ by L. lactis subsp. cremoris NZ9000 and their co-production with NisA by Lactococcus lactis subsp. lactis DPC5598 and L. lactis subsp. lactis BB24. The strains L. lactis subsp. cremoris WA2-67 (pJFQI), a producer of GarQ and NisZ, and L. lactis subsp. cremoris WA2-67 (pJFQIAI), a producer of GarA, GarQ and NisZ, demonstrated the highest antimicrobial activity (5.1- to 10.7-fold and 17.3- to 68.2-fold, respectively) against virulent L. garvieae strains. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae (L. garvieae) is a pathogenic gram-positive, catalase-negative (GPCN) bacterium that causes bovine mastitis. A total of 49 L. garvieae isolates were identified from 1441 clinical mastitis (CM) samples. The pathogenic effects of L. garvieae were studied with two infection models: bovine mammary epithelial cells cultured in vitro and murine mammary infections in vivo. The overall farm prevalence was 15.5% (13/84 farms in 9/19 provinces) and sample prevalence was 3.40% (49/1441). Post-treatment somatic cell count (SCC) post L. garvieae infection was significantly higher than the other GPCN pathogens isolated, and the bacteriological cure fraction was 41.94% (13/31) after intramammary antibiotic treatment. All L. garvieae isolates were resistant to rifaximin, 12.24% of isolates were resistant to cephalexin, and 10.20% (5/49) were multidrug-resistant (MDR). The most prevalent virulence genes were Hemolysin 1 (hly1)(100%), Hemolysin 2 (hly2) (97.96%), NADH oxidase (NADHO) (100%), Superoxide dismutase (SOD) (100%), Adhesin Pav (Pav) (100%), Adhesin PsaA (PsaA) (100%), Enolase (eno) (100%), Adhesin cluster 1(AC1) (100%), Adhesin cluster 2 (AC2) (100%), and several exopolysaccharides. L. garvieae rapidly adhered to bovine mammary epithelial cells, resulting in an elevated lactate dehydrogenase release. Edema and congestion were observed in challenged murine mammary glands and bacteria were consistently isolated at 12, 24, 48, 72, and 120 h after infection. We concluded that L. garvieae had good adaptive ability in the bovine and murine mammary cells and tissue. Given the resistance profile, penicillin and ampicillin are potential treatments for CM cases caused by L. garvieae. [ABSTRACT FROM AUTHOR]

ABSTRACT Lactococcus garvieae is an emerging zoonotic pathogen, but there are few reports regarding bovine mastitis. The prevalence of L. garvieae represents an increasing disease threat and global public health risk. Thirty-nine L. garvieae isolates were obtained from 2,899 bovine clinical mastitis milk samples in 6 provinces of China from 2017 to 2021. Five clonal complexes were determined from 32 multilocus sequence types (MLSTs) of L. garvieae: sequence type 46 (ST46) was the predominant sequence type, and 13 novel MLSTs were identified. All isolates were resistant to chloramphenicol and clindamycin, but susceptible to penicillin, ampicillin, amoxicillin-clavulanic acid, imipenem, ceftiofur, enrofloxacin, and marbofloxacin. Based on genomic analyses, L. garvieae had 6,310 genes, including 1,015 core, 3,641 accessory, and 1,654 unique genes. All isolates had virulence genes coding for collagenase, fibronectin-binding protein, glyceraldehyde-3-phosphate dehydrogenase, superoxide dismutase, and NADH oxidase. Most isolates had lsaD and mdtA antimicrobial resistance (AMR) genes. Based on COG (Clusters of Orthologous Genes database) results, the functions of defense, transcription and replication, and recombination and repair were enhanced in unique genes, whereas functions of translation, ribosomal structure, and biogenesis were enhanced in core genes. The KEGG functional categories enriched in unique genes included human disease and membrane transport, whereas COG functional categories enriched in core genes included energy metabolism, nucleotide metabolism, and translation. No gene was significantly associated with host specificity. In addition, analysis of core genome single nucleotide polymorphisms (SNPs) implied potential host adaptation of some isolates in several sequence types. In conclusion, this study characterized L. garvieae isolated from mastitis and detected potential adaptations of L. garvieae to various hosts. IMPORTANCE This study provides important genomic insights into a bovine mastitis pathogen, Lactococcus garvieae. Comprehensive genomic analyses of L. garvieae from dairy farms have not been reported. This study is a detailed and comprehensive report of novel features of isolates of L. garvieae, an important but poorly characterized bacterium, recovered in the past 5 years in 6 Chinese provinces. We documented diverse genetic features, including predominant sequence type ST46 and 13 novel MLSTs. Lactococcus garvieae had 6,310 genes, including 1,015 core, 3,641 accessory, and 1,654 unique genes. All isolates had virulence genes coding for collagenase, fibronectin-binding protein, glyceraldehyde-3-phosphate dehydrogenase, superoxide dismutase, and NADH oxidase and resistance to chloramphenicol and clindamycin. Most isolates had lsaD and mdtA antimicrobial resistance genes. However, no gene was significantly associated with host specificity. This is the first report that characterized L. garvieae isolates from bovine mastitis and revealed potential host adaptations of L. garvieae to various hosts.

Abstract This study aimed to evaluate the use of an oxy-hydrogen generator for preparing hydrogen-rich water (HRW) and studying its effects on the L. garvieae- affected zebrafish. 0, 10, 20, and 100% HRW were prepared to determine the effects of HRW on the mortality rate and gene expression levels of L. garvieae infected- zebrafish. After 48 hours of bacterial injection, the mortality rate of fish was 0, 0, 0, 66, 80, and 100% for the non-infected and non- HRW (G1), infected and 100% HRW (G5), non-infected and 100% HRW (G6), infected and 20% HRW (G4), infected and 10% HRW (G3), and infected without HRW (G2) groups. After 54 hours, there was a non-significant change in immunity-related gene expression levels (IL-1β and IL-6) between non-infected 100% HRW (G6) and non-infected non- HRW control (G1) groups. Gene expression levels were significantly upregulated for IL-1β (14, 13, and 9 times), IL-6 (48, 48, and 22 times), and SOD (9 times for each) genes in G2, G3, and G4 groups, respectively, but not for G5 comparing with control group G1. IL-1β and IL-6 gene expression levels decreased with the increase of hydrogen levels in the water. These results show that HRW could decrease (G3 and G4) and prevent (G5) the mortality of L. garvieae infected- zebrafish. This demonstrates the importance of the application of HRW for the treatment of bacterial infectious diseases of fish using a non-toxic compound as a green method without any residual effects on fish or water.

This mini review deals with some controversial non-starter lactic acid bacteria (NSLAB) species known to be both human and animal pathogens but also health-promoting and probiotic. The focus is on Lactococcus garvieae, two Streptococcus species (S. uberis and S. parauberis), four Weissella species (W. hellenica, W. confusa, W. paramesenteroides, and W. cibaria), and Mammalicoccus sciuri, which worldwide, are often found within the microbiotas of different kinds of cheese, mainly traditional artisanal cheeses made from raw milk and/or relying on environmental bacteria for their ripening. Based on literature data, the virulence and health-promoting effects of these bacteria are examined, and some of the mechanisms of these actions are reviewed. Additionally, their possible roles in cheese ripening are also discussed. The analysis of the literature data available so far showed that, in general, the pathogenic and the beneficial strains, despite belonging to the same species, show somewhat different genetic constitutions. Yet, when the safety of a given strain is assessed, genomic analysis on its own is not enough, and a polyphasic approach including additional physiological and functional tests is needed.

Lactococcus garvieae is a gram-positive cocci that has primarily been described as a pathogen in various fish species, but has increasingly been reported to cause endocarditis and other infections in humans [1]. Neonatal infection caused by Lactococcus garvieae has not been previously reported. Here we describe a premature neonate who developed a urinary tract infection with this organism and was successfully treated with vancomycin therapy. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae Lg‐per was originally isolated from rainbow trout cultured in cages located on the Turkish coast of the Black Sea in 2011. A whole genome sequence of Lg‐per was performed in the present study. The complete genome of Lg‐per mapped to the reference genomes of L. garvieae (GCF_000269925.1) and Lactococcus petauri (GCF_014830225.1) had a total of 1,694,407 and 1,945,297 base pairs, respectively. Lg‐per had 1955 protein‐coding genes and 4 rRNA, 46 tRNA and 1 tmRNA operons. The orthoANI value was 98.30% between Lg‐per and L. petauri (GCF_014830225.1) and 93.1% between Lg‐per and L. garvieae (GCF_000269925.1). A phylogenetic tree generated from the whole genome sequences (WGS) of several Lactococcus species found that L. petauri (GCA 002154895) was closely related to the Lg‐per strain with 98% similarity. Although L. garvieae Lg‐per was confirmed as L. garvieae based on phenotypical, biochemical and 16S rRNA sequence, WGS of the Lg‐per strain revealed that Lg‐per was L. petauri. Using a 16S rRNA‐based PCR detection approach, Lg‐per was misdiagnosed as L. garvieae since its 16S rRNA gene was 99.9% similar to that of L. garvieae strains. Consequently, the 16S rRNA‐based PCR detection approach may not be adequate for the identification of the Lactococcus genus. This is the first study to document the presence of L. petauri in Türkiye. L. garvieae isolates should be analysed using WGS since the same issue might occur in other countries. [ABSTRACT FROM AUTHOR]

BACKGROUND: There is growing evidence that resistance to antibiotics is increasing, and because of their wide range of negative impacts on the environment and humans, also accumulative effects in fish meat and hazards for humans, there have been numerous studies conducted using medicinal plants as a healthier alternative to antibiotics. OBJECTIVES: This study examined the antibacterial effect of two native medicinal herbs (oregano and echinacea mixed essential oil) on Lactococcus garvieae compared with some commercial antibiotics used in fish farms. METHODS: The antibacterial activities against a fish bacterial pathogen (Lactococcus garvieae) were tested using the disk diffusion method, followed by the determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations via sequential dilution of essential oil by broth micro dilution method. RESULTS: The results showed antimicrobial activities of mentioned mixed essential oil with ≥ 6.25 μL/mL and MBC ≥ 12.5 μL//mL. Also, the measurements of the diameter of inhibitory zones are consistent with the results of MIC and MBC, indicating the positive effect of this plant mix oil Lactococcus garvieae. However, the Lactococcus garvieae was more sensitive to enrofloxacin (NFX 5), and sulfamethoxazole & trimethoprim (SXT) (P<0.05), but the combination of oregano and echinacea essential oils performed better than florfenicol (FF) despite the weaker effects against mentioned two antibiotics. CONCLUSIONS: The results approved the antibacterial activity of the mixed essential oil of oregano and echinacea against Lactococcus garvieae on the laboratory scale. Using these compounds as a possible substitute for antibiotic compounds requires more clinical studies. [ABSTRACT FROM AUTHOR]

Lactococcus petauri is a recently described species of the genus Lactococcus. It was reported as an etiological agent of piscine lactococcosis together with Lactococcus garvieae. L. garvieae was already described as an opportunistic pathogen in human infections, with a potential zoonotic role. This paper represents the first report of a human urinary tract infection caused by L. petauri. A 91-year-old man was admitted to the emergency department for a femur fracture consequent to a domestic accident. The fracture was reduced by surgery and a catheterized specimen urine culture revealed a high bacterial load sustained by Gram-positive cocci, identified by Vitek 2 compact as L. garvieae, and subsequently as L. petauri through Internal Transcribed spacer 16S-23S r-RNA amplification. The number of L. petauri infections in humans is expected to rise in the near future mainly due to diagnostic improvement. A dedicated survey on L. garvieae and L. petauri infections in humans should be performed to better understand their role as pathogens and as zoonotic agents.

Introduction Lactococcus garvieae, a zoonotic pathogen, may rarely infect humans through the consumption of fish. Documented manifestations of L. garvieae infection in humans include infective endocarditis, prosthetic joint infections, liver abscesses, peritoneal dialysis-associated peritonitis, osteomyelitis, meningitis, infective spondylodiscitis, acalculous cholecystitis, and urinary tract infection. Case report An 87-year-old female was hospitalized for coffee-ground emesis secondary to acute gastritis after eating cooked fish. One week after her discharge, she developed new-onset confusion and was returned to the hospital. Chest computed tomography revealed total consolidation of the left lung and a multiloculated left pleural effusion. The patient required intubation and direct admission to the intensive care unit. Pleural fluid and blood cultures grew L. garvieae, which was susceptible to ceftriaxone, penicillin, and vancomycin. Despite intensive antibiotic therapy and supportive care for thirteen days, the patient remained in irreversible shock, and the family opted for comfort care. Conclusions Heretofore unreported, this case demonstrates that L. garvieae can cause bronchopneumonia and empyema. [ABSTRACT FROM AUTHOR]

Dietary polyphenol ellagic acid has anti-cancer and anti-inflammatory activities, and these biological activities require the conversion of ellagic acid to urolithins by intestinal microbes. However, few gut microbes are capable of metabolizing ellagic acid to produce urolithins, limiting the beneficial effects of ellagic acid on health. Here, we describe an intestinal bacterium Lactococcus garvieae FUA009 isolated from the feces of a healthy volunteer. It was demonstrated via HPLC and UPLC-MS analysis that the end product of ellagic acid metabolism of FUA009 was urolithin A. In addition, we also examined the whole genome sequence of FUA009 and then assessed the safety and probiotic properties of FUA009 based on a complete genome and phenotype analysis. We indicated that FUA009 was safe, which was confirmed by FUA009 being sensitive to multiple antibiotics, having no hemolytic activity, and being free of aggressive putative virulence factors. Moreover, 19 stress-responsive protein genes and 8 adhesion-related genes were predicted in the FUA009 genome. Furthermore, we demonstrated that FUA009 was tolerant to acid and bile salt by determining the cell viability in a stress environment. In summary, Lactococcus garvieae FUA009, as a novel UA-producing bacterium, not only contributes to the study of the metabolic pathway of ellagic acid but is also expected to be a novel probiotic candidate. [ABSTRACT FROM AUTHOR]

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The gut microbiota, a pivotal component of the intestinal mucosal barrier, is critical for host resistance to enteric pathogen infection. Here, we report a novel function of the potentially probiotic Lactococcus garvieae strain LG1 ( L. garvieae strain LG1) in maintaining intestinal mucosal barrier integrity and protecting against foodborne Clostridium perfringens ( C. perfringens ) infection. L. garvieae was isolated from the intestinal contents of Chinese Mongolian sheep (MS) and exhibited potential probiotic properties. In a C. perfringens enterocolitis model, L. garvieae- pretreated mice were less susceptible to C. perfringens infection compared with Phosphate buffered solution (PBS) - pretreated mice, which manifested as higher survival rates, lower pathogen loads, less weight loss, mild clinical symptoms and intestinal damage, and minor inflammation. Further mechanistic analysis showed that L. garvieae could ameliorate the disruption of intestinal permeability and maintain the integrity of the intestinal mucosal barrier by promoting the expression of tight junction proteins and mucoproteins. Moreover, L. garvieae was also able to facilitate antimicrobial peptide expression and ameliorate dysbiosis of the gut microbiota caused by C. perfringens . Together, these findings highlight the prospect of immunomodulatory potentially probiotic L. garvieae and might offer valuable strategies for prophylaxis and/or treatment of pathogenic C. perfringens mucosal infection., Importance: C. perfringens necrotic enteritis leads to losses of about US $2 billion to the poultry industry worldwide every year. Worse, US Centers for Disease Control and Prevention (CDC) has estimated that C. perfringens causes nearly 1 million foodborne illnesses in the United States annually. Nowadays, the treatment recommendation is a combination of a broad-spectrum synergistic penicillin with clindamycin or a carbapenem, despite growing scientific concern over antibiotic resistance. The global understanding of the gut microbiome for C. perfringens infection may provide important insights into the intervention. L. garvieae originated from Mongolian sheep intestine, exhibited potentially probiotic properties, and was able to limit C. perfringens enterocolitis and pathogenic colonization. Importantly, we found that L. garvieae limits C. perfringens invasion via improving intestinal mucosal barrier function. Also, L. garvieae alleviates C. perfringens -induced gut microbiota dysbiosis. It allowed us to convince that utilization of probiotics to promote protective immunity against pathogens infection is of pivotal importance., Competing Interests: The authors declare no conflict of interest.

The complete genome sequence of Lactococcus garvieae KN22525, isolated in May 2022 from a diseased greater amberjack ( Seriola dumerili ) with a gross indication of body surface redness in Nomi Bay, Japan, was determined. Multilocus sequencing typing revealed that KN22525's genotype was sequence type ST95., Competing Interests: The authors declare no conflict of interest.

The genus Lactococcus is a gram-positive, catalase-negative, non-motile bacterium. It is a facultative anaerobe and can be cultured at 10∼40°C. The genus Lactococcus consists of 16 species, of which Lactococcus garvieae and Lactococcus lactis are known to cause disease in humans. This study reports the first case in which L. garvieae was identified in the urine culture of a 74-year-old woman. The patient confirmed the findings of acute urinary tract infection through blood tests, microbial identification tests, antibiotic susceptibility tests, and computed tomography performed at a hospital. The patient was admitted to the nephrology ward and was treated with IV fluids and erythromycin antibiotics and discharged 5 days later. This is the first case in Korea in which L. garvieae was isolated from the urine of a patient with a urinary tract infection and is expected to be useful in treating patients with L. garvieae infection in the future.

Lactococcus garvieae is the etiological agent of lactococcosis, a clinically and economically significant infectious disease affecting farmed rainbow trout. L. garvieae had been considered the only cause of lactococcosis for a long time; however, L. petauri, another species of the genus Lactococcus, has lately been linked to the same disease. The genomes and biochemical profiles of L. petauri and L. garvieae have a high degree of similarity. Traditional diagnostic tests currently available cannot distinguish between these two species. The aim of this study was to use the transcribed spacer (ITS) region between 16S rRNA and 23S rRNA as a potential useful molecular target to differentiate L. garvieae from L. petauri, saving time and money compared to genomics methods currently used as diagnostic tools for accurate discrimination between these two species. The ITS region of 82 strains was amplified and sequenced. The amplified fragments varied in size from 500 to 550 bp. Based on the sequence, seven SNPs were identified that separate L. garvieae from L. petauri. The 16S-23S rRNA ITS region has enough resolution to distinguish between closely related L. garvieae and L. petauri and it can be used as a diagnostic marker to quickly identify the pathogens in a lactococcosis outbreak.

Lactococcus garvieae is the etiological agent of Lactococcosis, an evolving disease affecting many fish species and causing significant economic losses worldwide. Assessing pathogen relatedness and bacterial population structure is critical for determining the epidemiology of L. garvieae infections and in establishing effective pathogen management methods. The previously published morphological and genetic studies point to a clonal population structure, as seen in other fish bacteria. In the present study, the pulsed‐field gel electrophoresis (PFGE) method was utilized to define a population of 41 Taiwanese isolates from outbreaks with comparisons to four well‐characterized non‐Taiwanese isolates previously published. Two restriction enzymes (ApaI and SmaI) were utilized individually for PFGE analysis (cut‐off value = 90.0%), revealing genetic heterogeneity across L. garvieae isolates, with ApaI and SmaI yielding 12 and seven distinct PFGE band patterns, respectively. The phylogenic analysis using internal transcribed spacer region clustered all L. garvieae isolates in the same clad. Furthermore, the electron microscopic results confirmed the absence of capsular gene cluster (CGC) in previously characterized Taiwanese vaccine strain (S3) from grey mullet. Overall, our findings emphasize the importance of analysing the morphological and genetic diversity in L. garvieae being correlated for proper taxonomic classification in vaccine strain selection and epidemiological studies. [ABSTRACT FROM AUTHOR]

Lactococcus garvieae is an important pathogen of fish, associated with high rates of mortality and infection recurrence in summer or stressful conditions. Chronic infection and disease recurrence have also been reported to be associated with biofilms. However, the impact of biofilm and planktonic bacterial infection on fish immune responses remains unclear. In this study, de novo sequencing was used to compare differences of the spleen transcriptome in planktonic- and biofilm-infected mullets. Among the 181,024 unigenes obtained, 3,392 unigenes were associated with immune response genes. Comparative analysis of the gene expression between infection with the L. garvieae planktonic type and biofilm type identified a total of 3,120 and 3,489 differentially expressed genes in response to planktonic and biofilm infection, respectively, of which 1,366 and 1,458 genes were upregulated, and 1,754 and 1,458 genes were downregulated, respectively. Gene ontology enrichment analysis of immune genes identified genes involved in the complement system, toll-like receptor signaling, and antigen processing, which were further verified by qPCR. Additionally, genes encoding TLR2, IL-1β, TNF-α, C7, and MHC class II peptides were downregulated in response to biofilm infection. Importantly, the results show that biofilm infection induces a different immune pathway response compared with planktonic bacterial infection and, furthermore, illustrates that the prevention of biofilm formation may be a necessary and new strategy for controlling bacterial infection in aquaculture. [ABSTRACT FROM AUTHOR]