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Antimicrobial-resistant bacteria (ARB) from the intestinal contents of wild fish may have a relevant ecological significance and could be used as indicators of antimicrobial-resistance dissemination in natural bacterial populations in water bodies impacted by urban contamination. Thus, the occurrence of ARB in the intestinal contents of pelagic and demersal wild fishes captured in anthropogenic-impacted Coquimbo Bay in Chile was studied. Culturable counts of total and antimicrobial-resistant bacteria were determined by a spread plate method using Trypticase soy agar and R2A media, both alone and supplemented with the antimicrobials amoxicillin, streptomycin, florfenicol, oxytetracycline and ciprofloxacin, respectively. Heterotrophic plate counts of pelagic and demersal fishes ranged from 1.72 × 106 CFU g−1 to 3.62 × 109 CFU g−1, showing variable proportions of antimicrobial resistance. Representative antimicrobial-resistant isolates were identified by 16S rRNA gene sequencing, and isolates (74) from pelagic fishes mainly belonged to Pseudomonas (50.0%) and Shewanella (17.6%) genera, whereas isolates (68) from demersal fishes mainly belonged to Vibrio (33.8%) and Pseudomonas (26.5%) genera. Antimicrobial-resistant isolates were tested for susceptibility to 12 antimicrobials by an agar disk diffusion method, showing highest resistance to streptomycin (85.2%) and amoxicillin (64.8%), and lowest resistance to oxytetracycline (23.2%) and ciprofloxacin (0.7%). Only furazolidone and trimethoprim/sulfamethoxazole were statistically different (p < 0.05) in comparisons between isolates from pelagic and demersal wild fishes. Furthermore, an important number of these isolates carried plasmids (53.5%) and produced Extended-Spectrum-β-lactamases (ESBL) (16.9%), whereas the detection of Metallo–β–Lactamases and class 1-integron was rare. This study provides evidence that wild fish are important reservoirs and spreading-vehicles of ARB, carrying plasmids and producing ESBLs in Chilean marine environments. [ABSTRACT FROM AUTHOR]

Flavobacterial infection associated with diseased fish is caused by multiple bacterial species within the family Flavobacteriaceae. In the present study, the Chilean isolate FP99, from the gills of a diseased, farmed rainbow trout (Oncorhynchus mykiss), was characterized using phenotypic and genomic analyses. Additionally assessed was pathogenic activity. Phylogenetic analysis based on 16S rRNA gene sequencing confirmed that isolate FP99 belonged to the genus Epilithonimonas, an average nucleotide identity value of 100% was detected with the Chilean isolate identified as Epilithonimonas sp. FP211‐J200. In silico genome analysis, mechanisms for toxins production, and superantigens, adhesion, or other genes associated with virulence were not detected. However, genes encoding proteins for antibiotic resistance were found, including the chrA gene and the nucleotide sequence that encodes for multiple antibiotic resistance MarC proteins. Furthermore, the blaESP‐1 gene (87.85% aminoacidic sequence identity), encoding an extended‐spectrum subclass B3 metallo‐β‐lactamase and conferring carbapenem‐hydrolysing activity, and the tet(X) gene, which encodes a monooxygenase that catalyses the degradation of tetracycline‐class antimicrobials were carried by this isolate. Phenotyping analyses also supported assignment as E. ginsengisoli. Challenge trials against healthy rainbow trout resulted in no observed pathogenic effect. Our findings identify for the first time the species E. ginsengisoli as associated with fish farming, suggesting that this isolate may be a component of the microbiota of the freshwater system. Notwithstanding, poor environmental conditions and any stressors associated with aquaculture situations or lesions caused by other pathogenic bacteria, such as F. psychrophilum, could favour the entry of E. ginsengisoli into rainbow trout. [ABSTRACT FROM AUTHOR]

Two Vibrio strains (VPAP36 and VPAP40) were isolated from moribund-settled larvae of the Chilean scallop Argopecten purpuratus during vibriosis outbreaks that occurred in two commercial scallop larvae hatcheries located in the Inglesa and Tongoy bays in Northern Chile. The strains were identified as Vibrio chagasii using phenotypic characterization and whole genome sequence analysis. Both strains exhibited the phenotypic properties associated with virulence, gelatin hydrolysis and β-hemolysis, whereas only VPAP36 produced phospholipase and only VPAP40 produced caseinase. The whole genome analysis showed that the strains harbored genes encoding for the virulence factors, the EPS type II secretion system, and Quorum Sensing (auto-inductor 1 and auto-inductor 2), whereas genes encoding a metalloproteinase and a capsular polysaccharide were detected only in the VPAP40 genome. When challenge bioassays using healthy 11-day-old scallop larvae were performed, the V. chagasii VPAP36 and VPAP40 strains exhibited significant (p < 0.05) differences in their larval lethal activity, producing, after 48 h, larval mortalities of 65.51 ± 4.40% and 28.56 ± 5.35%, respectively. Otherwise, the cell-free extracellular products of the VPAP36 and VPAP40 strains produced larval mortalities of 20.86 ± 2.40% and 18.37 ± 2.40%, respectively, after 48 h of exposure. This study reports for the first time the isolation of V. chagasii from the massive larval mortalities of the farmed scallop (Argopecten purpuratus) in Chile, and demonstrates the pathogenic activity of V. chagasii towards the Chilean scallop, the second most important species for Chilean mariculture. [ABSTRACT FROM AUTHOR]

Chile is the second largest producer of salmon (only behind Norway), and high amounts of antimicrobials are used during the marine stage, mainly florfenicol. Intensive antimicrobials use is a relevant concern, prompting the urgent need for use reduction and advancements towards more restricted, efficient application. This review describes the historic and current use of antimicrobials in Chilean salmonid farming. Further covered is the experimental evidence reported to date, thus providing a clearer understanding for the pathogenic nature of Piscirickettsia salmonis—the most important bacterial pathogen and the main target of antimicrobials used in the Chilean industry. This review also assesses research exploring the impacts of antimicrobial treatment on the environment, as well as investigations on the antibiotic susceptibility of salmonid pathogens. This review of the scientific literature contributes to a more unbiased appraisal of the current situation in Chile. Unfortunately, various previous reports dealing with/on the antimicrobial use in the Chilean salmon farming industry have been arrived to insufficiently supported conclusions, thus the present review aims to clarify outdated or poorly substantiated information and assumptions in existing literature. The consequences of antimicrobials in fish farming are covered by this review, which is particularly relevant considering potential impacts on aquatic environments. Focus is given to the selection and spread of drug‐resistant bacteria and associated resistance‐encoding genes. Finally, herein are provided several recommendations from a One Health perspective for decreasing amounts of used antimicrobials, effectively controlling primary bacterial fish pathogens, developing proper prevention strategies and reducing the dissemination of antimicrobial resistance. [ABSTRACT FROM AUTHOR]

The VPAP30 strain was isolated as the highly predominant bacteria from an episode of massive larval mortality occurring in a commercial culture of the Chilean scallop Argopecten purpuratus. The main aims of this study were, to characterize and identify the pathogenic strain using biochemical and molecular methods, to demonstrate its pathogenic activity on scallop larvae, to characterize its pathogenic properties and to describe the chronology of the pathology. The pathogenic strain was identified as Vibrio bivalvicida based on its phenotypic properties, the multilocus sequence analysis (MLSA) of eight housekeeping genes (fts Z, gap A, gyr B, mre B, pyr H, rec A, rpo A, and top A) and different in silico genome-to-genome comparisons. When triplicate cultures of healthy 10 days old scallop larvae were challenged with 1 × 105 colony forming units (CFU) mL-1 of the VPAP30 strain, percentages of larval survival of 78.9 ± 3.3%, 34.3 ± 4.9%, and 0% were observed at 12, 2,4 and 36 h, respectively, whereas uninfected larval cultures showed survival rates of 97.4 ± 1.2% after of 48 h. Clinical symptoms exhibited by the scallop larvae infected with the VPAP30 strain include the accumulation of bacteria around the scallop larvae, velum disruption and necrosis of digestive gland. The 50% lethal dose (LD50) of VPAP30 strain at 24 and 48 h was 1.3 × 104 and 1.2 × 103 CFU mL-1, respectively. The invasive pathogenic activity of the VPAP30 strain was investigated with staining of the bacterial pathogen with 5-DTAF and analyzing bacterial invasion using epifluorescence, and a complete bacterial dissemination inside the larvae at 24 h post-infection was observed. When scallop larvae were inoculated with cell-free extracellular products (ECPs) of VPAP30, the larval survival rate was 59.5 ± 1.7%, significantly (P < 0.001) lower than the control group (97.4 ± 1.2%) whereas larvae treated with heat-treated ECPs exhibited a survival rate of 61.6 ± 1.8% after 48 h of exposure. V. bivalvicida VPAP30 exhibits high pathogenic activity on scallop larvae, mediated both by bacterial invasion and the production of toxigenic heat-stable compounds. This report constitutes the first isolation of V. bivalvicida out of Europe and extends the host range of this species, having demonstrated its pathogenic activity on the Chilean scallop larvae (A. purpuratus). These results supporting the pathogenic potential of V. bivalvicida to kill the larvae of a broad range of bivalve species reared in hatcheries located in the Atlantic and the Pacific coasts. [ABSTRACT FROM AUTHOR]

Three strains (VPAP16, VPAP18 and VPAP23 strains) were isolated as the most predominant organisms from 3 different episodes of massive mortalities of larval cultures of the Chilean scallop Argopecten purpuratus occurred in different commercial hatcheries located in northern Chile. The main aims of this study were to identify the pathogenic strains and investigate their pathogenic activity. Based on selected phenotypic features and sequence identity of the 16S rRNA gene and the housekeeping gene, RNA polymerase α-chain rpoA , all pathogenic strains were identified as Vibrio splendidus . Healthy 10 – day – old scallop larvae cultures exhibited mortality percentages of 69.61 ± 3.35%, 79.78 ± 6.11% and 61.73 ± 3.71% after 48 h when were inoculated with 1 × 10 6 CFU (colony forming units) mL −1 of VPAP16, VPAP18 and VPAP23 strains, respectively, and evidenced that concentrations ⩾10 4 CFU mL −1 would probably be detrimental for the larval culture. The main clinical signs observed in challenged larvae for 24 h were bacterial swarms on the margins of the larvae, extension and disruption of the velum, detachment of velum cilia cells and digestive tissue necrosis. Otherwise, challenge assays using pathogenic strains stained with 5-([4,6-dichlorotriazin-2-yl]amino)fluorescein hydrochloride (5-DTAF) evidenced that after 1 h stained bacteria were detected in high density in the digestive gland and the margin of the shell. When larval cultures were inoculated with cell-free extracellular products (ECP) of V. splendidus strains, exhibited larval mortalities higher than 70% (VPAP16), 80% (VPAP18) and 50% (VPAP23) after 24 h, even when ECP were treated with proteinase K or heat, indicating that extracellular pathogenic activity is mainly mediated by non-proteic thermostable compounds. In this study all Koch’s postulates were fulfilled and it was demonstrated for the first time the pathogenic activity of V. splendidus strains on reared-larvae of scallop A. purpuratus and prompt the necessity to maintain this species at concentrations lower than 10 4 CFU mL −1 to avoid episodes of mass mortalities in scallop hatcheries. [ABSTRACT FROM AUTHOR]

Abstract: In Chile, the administration of florfenicol to control bacterial pathologies in freshwater salmon farms is a frequent practice, but no studies has been developed to evaluate the incidence of florfenicol-resistant bacteria in these cultures. This study was undertaken to obtain information on the occurrence of florfenicol-resistant bacteria in a farm (LF1) under florfenicol therapy two weeks before sampling date, located in the Rupanco Lake, and a farm (LF2) with no recent history of antibacterial therapy, located in the Llanquihue Lake. Samples from surface water, pellet, Salmo salar fingerlings and control and under-cage sediments were collected from each salmon farm. Total and florfenicol-resistant culturable counts were determined by a spread plate method, observing low (<9%) proportions of florfenicol resistance in water and fingerling samples, whereas sediment samples from LF1 farm showed proportions ranging from 18.55±2.85% (control) to 26.40±1.56% (under-cage) and were significantly higher (P <0.05) than those from LF2 farm, with 0.29±0.30% (control) and 0.69±1.00% (under-cage) values, not observing a direct correlation with sediment organic enrichment. Also, unmedicated pelletized feed from LF1 farm showed high levels of resistance (34.79±11.87%), and were significantly (P <0.05) higher than those from LF2 farm (<0.001%). A number of 34 and 36 representative isolates recovered from LF1 and LF2 farms respectively, were selected and identified by using the Biolog Microbial ID System, observing that mainly belonged to Pseudomonas genus (67.65 and 66.67% in LF1 and LF2 farms) and the predominant species were P. putida and P. viridilivida (14.71%) in LF1 farm, and P. fluorescens (36.11%) in LF2 farm. Antibacterial susceptibility patterns of resistant strains from both Lakes were similar, showing important proportions of isolates resistance to ampicillin, erythromycin, furazolidone and cotrimoxazole and susceptibility to gentamicin, kanamycin and enrofloxacin. The minimum inhibitory concentration (MIC) of florfenicol against these selected bacteria were determined, observing MIC ranges of 256–>1024 and 64–>1024 μg ml−1 for strains recovered from LF1 and LF2 farms, respectively. The important occurrence of florfenicol-resistant and antibacterial-multiresistant microorganisms in unpolluted and farm-impacted sites in Rupanco Lake suggests that use of florfenicol is not a necessary causal condition for the development of elevated frequencies of florfenicol resistance. Otherwise, considering that intrinsic resistance to many antibacterials has been extensively reported for Pseudomonas species, observed levels of resistance to florfenicol could be overestimated by the occurrence of innately resistant bacteria with multiple non-specific resistances. Our results prompt the necessity for maintaining a continuous surveillance of florfenicol resistance in salmon farms, as well as to characterize the molecular elements involved in these resistances to evaluate the feasibility of their dissemination among environmental microbiota. [Copyright &y& Elsevier]

The use of oxytetracycline for preventing and controlling bacterial pathogens in salmon farming is frequent in Chile, yet no studies have been performed to evaluate the ecological impact of its intensive and prolonged use. In this work, the frequency of oxytetracycline-resistant bacteria from water, pelletized feed and fingerlings from four Chilean freshwater Atlantic salmon farms, as well as the level of resistance of selected strains was investigated. Viable bacterial counts were performed by a spread plate method and antibiotic-resistant bacteria were counted in culture media supplemented with the selected antibiotic. Resistance levels of selected strains isolated in media containing antibiotic were determined using an agar plate dilution method. High proportions of low- and high-level oxytetracycline-resistant bacteria (selected in agar plates containing 30 and 100 μg/ml, respectively), mainly from pelletized feed and effluent samples of the fish farms were found. The highest proportions of resistant bacteria were found in the effluent samples, and were significantly higher (P<0.05, Tukey''s test) than those from the other samples studied. On the contrary, influent samples exhibited the lowest proportions of resistant bacteria. One hundred and three resistant Gram-negative isolates, which represented the oxytetracycline-resistant bacterial population, were randomly selected on TSA containing 30 or 100 μg/ml of oxytetracycline, from salmon farms and pellet samples, and streaked for purification on TSA plates without oxytetracycline. A large number of non-fermenting bacteria (77.7%) were identified. Among these, Pseudomonas fluorescens (28.2%), Stenotrophomonas maltophilia (5.8%), Sphingomonas paucimobilis (5.8%), Acinetobacter lwoffii (4.8%), and Pseudomonas putida (4.8%) were the most frequent. Also, an important number of strains of Aeromonas hydrophila (9.7%), Burkholderia cepacia (3.9%), Brevundimonas vesicularis (3.9%), Acinetobacter johnsonii (2.9%), Pantoea sp. (2.9%) and Moraxella sp. (2.9%) were found. P. fluorescens and A. hydrophila predominated in salmon fingerlings, whereas A. lwoffii and S. maltophilia were predominant in pellet samples. Selected strains exhibited high levels of oxytetracycline resistance, with minimum inhibitory concentrations (MICs) ranging from 64 to 2048 μg/ml, whereas MIC90 of oxytetracycline varied between 1024 and 2048 μg/ml. This study shows the presence of an important population of oxytetracycline-resistant bacteria in the microflora of Chilean salmon farms. Therefore, the environment of these farms might play important roles as reservoirs of bacteria carrying genetic determinants for high-level tetracycline resistance, prompting an important risk to public health for workers involved in fish culturing and processing. [Copyright &y& Elsevier]

The intensive use of antimicrobial agents, mainly oxytetracycline, to prevent and control bacterial pathologies in Chilean salmon culture is a frequent practice. A total of 103 Gram-negative oxytetracycline-resistant bacteria recovered from various sources of 4 Chilean freshwater salmon farms were identified and investigated for their susceptibility patterns to various antibacterial agents, by using an agar disk diffusion method. Antibacterial resistance patterns of isolates were not correlated with bacterial species or strain source. A high number of bacteria resistant to amoxicillin, ampicillin, erythromycin, and furazolidone, as well as an important frequency of bacterial resistance to florfenicol, chloramphenicol, cefotaxime and trimethoprim-sulfamethoxazole was found. On the contrary, the proportion of bacteria resistant to gentamicin, kanamycin, flumequine and enrofloxacin was rather low. Resistant microflora showed a high taxonomic variability and mainly consisted of non-fermenting bacteria (77.7%). These strains mainly belonged to the species Pseudomonas fluorescens (29), Aeromonas hydrophila (10), Stenotrophomonas maltophilia (6), isolated from salmon fingerlings, and Acinetobacter lwoffii (5) isolated from pelletized feed. The occurrence of simultaneous resistance to various antibacterials was frequent. We observe a high frequency of bacteria resistant to 6–10 antibacterials (74 strains), and antibiotic resistance index (ARI) values ranging from 0.38 to 0.48 for the four salmon farms studied. These results suggest that Chilean salmon farms might play a role as reservoirs of antibacterial multiresistant bacteria, thus prompting the necessity for a more restrictive attitude towards the intensive use of antibacterials in salmon farming. [Copyright &y& Elsevier]

Chile has promoted the diversification of aquaculture and red cusk-eel (Genypterus chilensis) is one of the prioritized species. However, many aspects of the biology of the species are unknown or have little information available. These include intestinal microbiota, an element that may play an important role in the nutrition and defense of cultured animals for meat production. This study compares the microbiota composition of the intestinal contents of wild and aquaculture fish to explore the microbial communities present and their potential contribution to the host. DNA was extracted from the intestinal content samples and the V4 region of the 16S rRNA gene was amplified and sequenced using the Ion Torrent platform. After the examination of the sequences, strong differences were found in the composition at the level of phylum, being Firmicutes and Tenericutes the most abundant in aquaculture and wild condition, respectively. At the genus level, the Vagococcus (54%) and Mycoplasma (97%) were the most prevalent in the microbial community of aquaculture and wild condition, respectively. The evaluation of predicted metabolic pathways in these metagenomes showed that in wild condition there is an important presence of lipid metabolism belonging to the unsaturated fatty acid synthesis. In the aquaculture condition, the metabolism of terpenoids and polyketides were relevant. To our knowledge, this is the first study to characterize and compare the intestinal microbiota of red cusk-eel (Genypterus chilensis) of wild and aquaculture origin using high-throughput sequencing. [ABSTRACT FROM AUTHOR]

The main objective of this study was to characterize the tet(X) genes, which encode a monooxygenase that catalyzes the degradation of tetracycline antibiotics, carried by the resistant strains FP105 and FP233-J200, using whole-genome sequencing analysis. The isolates were recovered from fin lesion and kidney samples of diseased rainbow trout Oncorhynchus mykiss, during two Flavobacteriosis outbreaks occurring in freshwater farms located in Southern Chile. The strains were identified as Epilithonimonas spp. by using biochemical tests and by genome comparison analysis using the PATRIC bioinformatics platform and exhibited a minimum inhibitory concentration (MIC) of oxytetracycline of 128 µg/mL. The tet(X) genes were located on small contigs of the FP105 and FP233-J200 genomes. The sequences obtained for the tet(X) genes and their genetic environment were compared with the genomes available in the GenBank database of strains of the Chryseobacterium clade belonging to the Flavobacterium family, isolated from fish and carrying the tet(X) gene. The Tet(X) proteins synthesized by the Chilean Epilithonimonas strains showed a high amino acid similarity (range from 84% to 100%), with the available sequences found in strains belonging to the genus Chryseobacterium and Flavobacterium isolated from fish. An identical neighborhood of tet(X) genes from both Chilean strains was observed. The genetic environment of tet(X) observed in the two strains of Epilithonimonas studied was characterized by the upstream location of a sequence encoding a hypothetical protein and a downstream located alpha/beta hydrolase-encoding gene, similar to the observed in some of the tet(X) genes carried by Chryseobacterium and Flavobacterium strains isolated from fish, but the produced proteins exhibited a low amino acid identity (25–27%) when compared to these synthesized by the Chilean strains. This study reports for the first time the carriage of the tet(X) gene by the Epilithonimonas genus and their detection in fish pathogenic bacteria isolated from farmed salmonids in Chile, thus limiting the use of therapies based on oxytetracycline, the antimicrobial most widely used in Chilean freshwater salmonid farming. This results suggest that pathogenic strains of the Chryseobacterium clade occurring in Chilean salmonid farms may serve as important reservoirs of tet(X) genes. [ABSTRACT FROM AUTHOR]

The main objective of this study was to characterize using whole-genome sequencing analysis, a new variant of the qnrB gene (qnrB89) carried by a fluoroquinolone-susceptible bacterium isolated from mucus of farmed Salmo salar fingerling in Chile. Citrobacter gillenii FP75 was identified by using biochemical tests and 16S ribosomal gene analysis. Nucleotide and amino acid sequences of the qnrB89 gene exhibited an identity to qnrB of 81.24% and 91.59%, respectively. The genetic environment of qnrB89 was characterized by the upstream location of a sequence encoding for a protein containing a heavy metal-binding domain and a gene encoding for a N-acetylmuramoyl-L-alanine amidase protein, whereas downstream to qnrB89 gene were detected the csp and cspG genes, encoding cold-shock proteins. The qnrB89 gene was located on a large chromosomal contig of the FP75 genome and was not associated with the 10-kb plasmid and class 1 integron harbored by the FP75 strain. This study reports for the first time the carriage of a qnrB gene by the C. gillenii species, and its detection in a bacterial strain isolated from farmed salmon in Chile. [ABSTRACT FROM AUTHOR]

Flavobacteriosis, caused by Flavobacterium psychrophilum is considered the most important bacterial disease in Chilean freshwater salmonid farming, and is currently mainly controlled by the use of antimicrobial agents. The aim of this study was to correlate the results obtained by disk diffusion and minimum inhibitory concentration (MIC) assays of antimicrobials used in fish farming using F. psychrophilum isolates recovered from Chilean farmed salmonids. Antimicrobial susceptibility patterns and MIC values of six antibacterial agents against 125 Chilean isolates of F. psychrophilum were tested on Diluted (1:5) Cation-Adjusted Mueller-Hinton (DCAMH) agar using a disk diffusion method and an agar dilution method, respectively. A moderate correlation for antimicrobials amoxicillin and florfenicol was observed (Pearson correlation indices of −0.6894 and −0.5857, respectively), whereas highly significantly high (p <.001) correlations were detected for oxytetracycline, oxolinic acid, flumequine and enrofloxacin (Pearson correlation indices of −0.7901, −0.8386, −0.8151 and −0.7818, respectively), demonstrating that MIC and disk values were tightly associated (R2 ≥ 0.61). When isolates were categorized as fully susceptible Wild-Type or Non Wild-Type, the number of isolates differently categorized using each method varied between 8 and 23 for assayed antibacterials. The use of DCAMH without biological supplements was efficient to perform agar dilution tests with F. psychrophilum but its use to perform disk diffusion assays is questionable, due to some isolates exhibiting poor growth produced inhibition zones that are difficult to read. This study demonstrates that the agar dilution method is a good alternative to determine MIC values when a high number of isolates must be assayed. In contrast, further research is required to advance in the standardization of a disk diffusion procedure for this pathogen. • Disk diffusion and MIC values of oxytetracycline and quinolones of F. psychrophilum were significantly (p <.001) correlated • Lower correlations between disk diffusion and MIC values of amoxicillin and florfenicol of F. psychrophilum were shown. • Diluted Mueller-Hinton without biological supplements is efficient to perform agar dilution tests of Chilean F. psychrophilum. • Agar dilution method is a good alternative to determine MIC values when a high number of F. psychrophilum strains are assayed. [ABSTRACT FROM AUTHOR]

[Display omitted] • Our results demonstrated the global spread of the emergent pathogen V. europaeus. • V. europaeus was isolated in Spain, France, and now for first time in Chile and US. • Genome of V. europaeus has been fully resolved for first time. • We also showed that the V. europaeus host range includes A. purpuratus. • This species affected the most important species reared in bivalve hatcheries. Vibrio europaeus is an emergent pathogen affecting the most important bivalve species reared in Spanish and French hatcheries. Using a genomic approach, we identified V. europaeus outside Europe for the first time from massive larval mortalities of scallop (Argopecten purpuratus) in Chile and from seawater near a shellfish hatchery in the US West Coast. Results show the worldwide spreading and potential impact of V. europaeus for aquaculture; these four countries are among the 10 major producers of mollusks. Pathogenicity of V. europaeus was demonstrated for the first time towards scallop, the second most important species for Chilean mariculture. [ABSTRACT FROM AUTHOR]

Several members of the Mycobacterium genus cause invasive infections in humans and animals. According to a recent phylogenetic analysis, some strains of Mycobacterium salmoniphilum (Msal), which are the main culprit in bacterial outbreaks in freshwater fish aquaculture, have been assigned to a separate branch containing Mycobacterium franklinii (Mfra), another species that causes infections in humans. However, this genus is little studied in an aquaculture context. Here, we isolated four Mycobacterium spp. strains from freshwater cultures of Atlantic and coho salmon in Chile and performed whole-genome sequencing for deep genomic characterization. In addition, we described the gross pathology and histopathology of the outbreaks. Several bioinformatic analyses were performed using the genomes of these four Mycobacterium isolates in conjunction with those of Msal strains, four Msal-like strains, and one Mfra strains, plus 17 other publicly available Mycobacterium genomes. We found that three isolates are clustered into the Msal branch, whereas one isolate clustered with the Mfra/Msal-like strains. We further evaluated the presence of virulence and antimicrobial resistance genes and observed that the four isolates were closely related to the Msal and Msal-like taxa and carried several antimicrobial resistance and virulence genes that are similar to those of other pathogenic members of the Mycobacterium clade. Altogether, our characterization Msal and Msal-like presented here shed new light on the basis of mycobacteriosis provides quantitative evidence that Mycobacterium strains are a potential risk for aquaculture asetiological agents of emerging diseases, and highlight their biological scopes in the aquaculture industry., (© 2021. The Author(s).)

The interest in and demand for natural dyes has increased significantly in recent years; however, very few natural blue dyes are commercially available, because blue colored compounds in nature are relatively rare. In this study, a blue pigment-producing bacteria from Lake Chungará (Atacama Desert, Chile) was isolated, and its blue pigment was purified and chemically characterized. The pigment-producing strain was identified as Pseudarthrobacter sp. by 16S rRNA gene sequencing. The pigment was separated from the filtered culture medium by column chromatography/solid-phase extraction using different resins (ionic exchange, C-18, size exclusion). The strain produced up to 2.5 g L -1 of blue pigment, which was very soluble in water, partially soluble in methanol and insoluble in other organic solvents. The pigment was analyzed and characterized by analytical HPLC, UV-Vis, FT-IR, and H-NMR, and purified by semi-preparative HPLC. The pigment was non-toxic to brine shrimp (LD 50  > 2.3 g L -1 ) and was stable at pH 6-10 at temperatures below 60 °C. HPLC analysis shows that the pigment is composed of four major blue fractions. The physicochemical properties and structural analysis demonstrate that this pigment belongs to the indochrome isomers, whose properties have yet to have been characterized. The high solubility in water, good stability in neutral and basic pH, and negligible toxicity of the blue pigment make it a good candidate suitable for several industrial and possibly some food applications.

Mass mortalities of larval cultures of Chilean scallop Argopecten purpuratus have repeatedly occurred in northern Chile, characterized by larval agglutination and accumulation in the bottom of rearing tanks. The exopolysaccharide slime (EPS) producing CAM2 strain was isolated as the primary organism from moribund larvae in a pathogenic outbreak occurring in a commercial hatchery producing larvae of the Chilean scallop Argopecten purpuratus located in Bahía Inglesa, Chile. The CAM2 strain was characterized biochemically and was identified by polymerase chain reaction amplification of 16S rRNA as Halomonas sp. (Accession number DQ885389.1). Healthy 7-day-old scallop larvae cultures were experimentally infected for a 48-h period with an overnight culture of the CAM2 strain at a final concentration of ca. 10(5) cells per milliliter, and the mortality and vital condition of larvae were determined by optical and scanning electron microscopy (SEM) to describe the chronology of the disease. Pathogenic action of the CAM2 strain was clearly evidenced by SEM analysis, showing a high ability to adhere and detach larvae velum cells by using its "slimy" EPS, producing agglutination, loss of motility, and a posterior sinking of scallop larvae. After 48 h, a dense bacterial slime on the shell surface was observed, producing high percentages of larval agglutination (63.28 +/- 7.87%) and mortality (45.03 +/- 4.32%) that were significantly (P < 0.05) higher than those of the unchallenged control cultures, which exhibited only 3.20 +/- 1.40% dead larvae and no larval agglutination. Furthermore, the CAM2 strain exhibited a high ability to adhere to fiberglass pieces of tanks used for scallop larvae rearing (1.64 x 10(5) cells adhered per square millimeters at 24 h postinoculation), making it very difficult to eradicate it from the culture systems. This is the first report of a pathogenic activity on scallop larvae of Halomonas species, and it prompts the necessity of an appraisal on biofilm-producing bacteria in Chilean scallop hatcheries.

Twenty-five distinct tetracycline-resistant gram-negative bacteria recovered from four Chilean fish farms with no history of recent antibiotic use were examined for the presence of tetracycline resistance (tet) genes. Sixty percent of the isolates carried 1 of the 22 known tet genes examined. The distribution was as follows. The tet(A) gene was found in six isolates. The tet(B) gene was found in two isolates, including the first description in the genus Brevundimonas: Two isolates carried the tet(34) and tet(B) genes, including the first description of the tet(34) gene in Pseudomonas and Serratia and the first description of the tet(B) gene in Pseudomonas: The tet(H) gene was found in two isolates, which includes the first description in the genera Moraxella and Acinetobacter: One isolate carried tet(E), and one isolate carried tet(35), the first description of the gene in the genus Stenotrophomonas: Finally, one isolate carried tet(L), found for the first time in the genus Morganella: By DNA sequence analysis, the two tet(H) genes were indistinguishable from the previously sequenced tet(H) gene from Tn5706 found in Pasteurella multocida. The Acinetobacter radioresistens isolate also harbored the Tn5706-associated 1,063-bp IS element IS1597, while the Moraxella isolate carried a 1,026-bp IS-like element whose 293-amino-acid transposase protein exhibited 69% identity and 84% similarity to the transposase protein of IS1597, suggesting the presence of a novel IS element. The distribution of tet genes from the Chilean freshwater ponds was different than those that have previously been described from other geographical locations, with 40% of the isolates carrying unidentified tetracycline resistance genes.