Antimicrobial susceptibility of Peruvian Yersinia ruckeri isolates to oxytetracycline and florfenicol using MIC testing.

Yersiniosis, caused by Yersinia ruckeri, has become the most common disease in farmed rainbow trout Oncorhynchus mykiss in Peru, affecting Puno and Junín Regions, important aquaculture areas in the country. Florfenicol (FLO) and oxytetracycline (OXY) are the antimicrobials most frequently used to mitigate losses attributed to this pathogen. This study presents an analysis of the susceptibility patterns of 60 Y. ruckeri isolates (30 isolates each from Puno and Junín), including the type strain CECT 4319T and the strains CECT 955 and CECT 956, against FLO and OXY. Minimum inhibitory concentrations (MICs) were determined following the guideline for standard broth dilution method published by the Clinical and Laboratory Standards Institute. MIC results ranged from 4.0 to 8.0 µg ml-1 for FLO and 0.5 to 4.0 µg ml-1 for OXY. Normalized resistance interpretation (NRI) analysis identified epidemiological cut-off values of ≤16.0 µg ml-1 for FLO and ≤4.0 µg ml-1 for OXY. All Peruvian isolates, including the collection strains, were categorized as wild-type for both antimicrobials. Even though the number of Y. ruckeri isolates with MIC values of 8 µg ml-1 for FLO is more than double in Puno than in Junín (15 vs. 7 isolates), the NRI analysis showed the same epidemiological cutoff of 16 µg ml-1; while for OXY, it was 4.0 µg ml-1 for Puno and 2.0 µg ml-1 for Junín. This study establishes the basis for monitoring susceptibility to FLO and OXY in new Y. ruckeri isolates in Peruvian rainbow trout farming.

Insights into the genomic traits of Yersinia frederiksenii, Yersinia intermedia and Yersinia kristensenii isolated from diverse sources in Brazil.

Yersinia is an important genus comprising foodborne, zoonotic and pathogenic bacteria. On the other hand, species of the so-called group Yersinia enterocolitica-like are understudied and mostly characterized as non-pathogenic, despite of some reports of human infections. The present study aimed to provide genomic insights of Yersinia frederiksenii (YF), Yersinia intermedia (YI) and Yersinia kristensenii (YK) isolated worldwide. A total of 22 YF, 20 YI and 14 YK genomes were searched for antimicrobial resistance genes, plasmids, prophages, and virulence factors. Their phylogenomic relatedness was analyzed by Gegenees and core-genome multi-locus sequence typing. Beta-lactam resistance gene blaTEM-116 and five plasmids replicons (pYE854, ColRNAI, ColE10, Col(pHAD28) and IncN3) were detected in less than five genomes. A total of 59 prophages, 106 virulence markers of the Yersinia genus, associated to adherence, antiphagocytosis, exoenzymes, invasion, iron uptake, proteases, secretion systems and the O-antigen, and virulence factors associated to other 20 bacterial genera were detected. Phylogenomic analysis revealed high inter-species distinction and four highly diverse YF clusters. In conclusion, the results obtained through the analyses of YF, YI and YK genomes suggest the virulence potential of these strains due to the broad diversity and high frequency of prophages and virulence factors found. Phylogenetic analyses were able to correctly distinguish these closely related species and show the presence of different genetic subgroups. These data contributed for a better understanding of YF, YI and YK virulence-associated features and global genetic diversity, and reinforced the need for better characterization of these Y. enterocolitica-like species considered non-pathogenic.

Comparative genomics and antibiotic resistance of Yersinia enterocolitica obtained from a pork production chain and human clinical cases in Brazil.

Previous work found a high similarity of macro-restriction patterns for isolates of Yersinia enterocolitica 4/O:3 obtained at a pork production chain from Minas Gerais, Brazil. Herein we aimed to determine the clonality and the antibiotic resistance profiles of a subset of these isolates (n = 23) and human clinical isolates (n = 3). Analysis based on whole genome sequencing (WGS) showed that the isolates were distributed into two major clades based on single nucleotide polymorphisms (SNP) with one isolate defining Clade A (isolate R31) and remaining isolates (n = 25, 96.2%) defining Clade B. Seven clonal groups were identified. The inclusion of isolate R31 as a distinct clonal group was due to the presence of several phage-related genes, allowing its characterization as serotype O:5 by WGS. Disk-diffusion assays (14 antibiotics) identified 13 multidrug resistant isolates (50.0%). Subsequent sequence analysis identified 17 different antibiotic resistance related genes. All isolates harbored blaA (y56 beta-lactamase), vatF, rosA, rosB and crp, while nine isolates harbored a high diversity of antibiotic resistance related genes (n = 13). The close genetic relationship among Y. enterocolitica obtained from a pork production chain and human clinical isolates in Brazil was confirmed, and we can highlight the role of swine in the potential transmission of an antibiotic-resistant clones of a pathogenic bio-serotype to humans, or the transmission of these resistant bacteria from people to animals. The role of veterinary antibiotic use in this process is unclear.

Pathogenic potential and antibiotic resistance of Yersinia enterocolitica, a foodborne pathogen limited to swine tonsils in a pork production chain from Southern Brazil.

In this study, we aimed to characterize the distribution of Yersinia enterocolitica in a pork production chain in Brazil, as well as the virulence profile and antibiotic resistance of the obtained isolates. Samples from 10 pig lots obtained from finishing farms (water, feed, and barn floors, n = 30), slaughterhouse (lairage floors, carcasses at four processing steps, tonsils, and mesenteric lymph nodes, n = 610), and processing (end cuts, processing environment, n = 160) were obtained in Paraná state, Brazil, and subjected to Y. enterocolitica detection by ISO 10,273. The obtained isolates were identified based on biochemical and molecular features (16 s rRNA, inv, bioserotyping) and subjected to PCR assays to detect virulence (ail, ystA, ystB, virF, myfA, fepA, fepD, fes, tccC, ymoA, hreP, and sat) and multidrug resistance-related genes (emrD, yfhD, and marC). Also, isolates were subjected to disk diffusion test to characterize their resistance against 17 antibiotics from 11 classes and to pulsed field gel electrophoresis (PFGE) after XbaI macro-restriction. Y. enterocolitica was detected in a single sample (tonsil), and the obtained three isolates were characterized as serotype O:3, harboring ail, ystA, virF, myfA, tccC, ymoA, hreP, emrD, yfhD, and marC, and resistant to all tested antibiotics. The three isolates presented identical macro-restriction profiles by PFGE, also identical to isolates obtained from Minas Gerais, other Brazilian state; one selected isolate was identified as biotype 4. Despite the low occurrence of Y. enterocolitica in the studied pork production, the virulence potential and the antibiotic resistance profiles of the isolates demonstrated their pathogenic potential, and the macro-restriction profiles indicate strains descending from a common subtype in the pork production chain of two Brazilian States.

Persistence of Yersinia enterocolitica bio-serotype 4/O:3 in a pork production chain in Minas Gerais, Brazil.

Yersinia enterocolitica bio-serotype 4/O:3 was previously identified in a pork production chain in Brazil and the obtained isolates presented high identity by pulsed-field gel electrophoresis (PFGE, XbaI). For the current study, an additional 147 porcine samples (tonsils = 100, palate = 30, head meat = 17) were collected from the same pork production chain 2-years later and 14 (9.5%) tested positive for Y. enterocolitica. Isolates (n = 24, 1 to 2 per positive sample) were bio-serotype 4/O:3 and harbored virulence genes ail, inv, wbbU, virF, myfA, ystA, ymoA, hreP and sat, and the multidrug resistance related genes emrD, marC and yfhD. PFGE (XbaI) demonstrated no differences among isolates (100% similarity) and were identical to some Y. enterocolitica isolates (n = 13) obtained previously from the same pork chain. A second PFGE analysis (NotI) confirmed the high degree of similarity among isolates obtained over time, demonstrating the persistence of an apparent clonal Y. enterocolitica bio-serotype 4/O:3 in this particular pork production chain in Brazil.

An overview of Yersinia enterocolitica and related species in samples of different origin from San Luis, Argentina.

This study is aimed at offering an overview of the prevalence of Yersinia enterocolitica and related species in San Luis, Argentina, from samples of diverse origin received in our laboratory between 1984 and 2014, and providing an analysis of the distribution of Yersinia isolates according to their isolation sources, highlighting bioserotypes and potential reservoirs and vehicles of transmission to humans. From a total of 4572 samples of human, animal, food and environmental origins analyzed by traditional culture methods and molecular techniques, 229 (5%) samples were Yersinia positive. The highest frequency of Yersinia isolates was observed in environmental specimens (14.3%), followed by animal (9.2%), food (5%) and human (0.6%) samples. A total of 255 Yersinia isolates were characterized, including 183 Y. enterocolitica and 72 isolates of other Yersinia species. Biotype 1A associated to several serotypes was identified in Y. enterocolitica isolates from environment (100%), animals (95.5%), foods (71.7%) and human samples (40%); bioserotype 2/O:9 was identified in isolates from foods (25.5%), and biotype 3 was associated with strains from humans (60%), animals (4.5%) and foods (2.8%). This biotype included three strains O:3 and six strains O:5. The data highlight animals and foods as the main Y. enterocolitica sources in our region.

Aporphinoid alkaloids as antimicrobial agents against Yersinia enterocolitica.

Foodborne diseases have become a health issue worldwide, mainly due to the consumption of contaminated foods that are either raw, improperly heat treated or cross-contaminated after adequate heat treatment foods. A group of alkaloids extracted from plants were tested to evaluate their antimicrobial effect against different strains of Yersinia enterocolitica and other foodborne bacteria. The results obtained reveal that oliveridine and pachypodanthine inhibited Y. enterocolitica growth, with MIC values of 25 µmol l-1 and 100 µmol l-1 respectively. The results indicated that both alkaloids are good growth inhibitors, but oliveridine showed greater inhibitory effect with lower MIC values. Inhibitory alkaloids can be developed as potential antimicrobials in food system to prevent or treat foodborne diseases, thus contributing to solve the global issue of contaminated food consumption. SIGNIFICANCE AND IMPACT OF THE STUDY: Alkaloids are abundant secondary metabolites in plants and represent one of the most widespread class of compounds endowed with multiple and varied pharmacological properties. In this work, we propose two aporphinoid alkaloids extracted from plants as new antimicrobial agents. Oliveridine and pachypodanthine inhibited Yersinia enterocolitica growth for up to 96 h of culture. This is the first reported study of the activity of these alkaloids as antimicrobial compounds.

Yersinia enterocolitica in a Brazilian pork production chain: Tracking of contamination routes, virulence and antimicrobial resistance.

This study aimed to track Yersinia enterocolitica contamination in a pork production chain in Minas Gerais, Brazil, and to characterize the virulence and antibiotic resistance of isolates. Samples were collected from four different steps of the pork production chain (pig farm, carcass, processing environment and end product; n = 870), and tested for the presence of Y. enterocolitica. The pathogen was detected in 8 samples (palatine tonsils = 5; mesenteric lymph nodes = 2; carcass after bleeding = 1), from which 16 isolates were obtained and identified as Y. enterocolitica bioserotype 4/O:3. XbaI macrorestriction allowed the clustering of isolates in 5 pulsetypes, and the identification of identical profiles of Y. enterocolitca isolated from different samples. All isolates were positive for the virulence related genes ail, virF, myfA, ystA, tccC, ymoA, hreP and sat, and negative for ystB, ystC, fepA, fepD and fes. Considering 17 antibiotics from 11 classes, only ciprofloxacin and kanamycin were effective against all isolates, and three multidrug resistance profiles were identified among them, with simultaneous resistance to 9 of 11 classes. All isolates presented positive results for emrD, yfhD and marC, related to multidrug resistance. The results of this study demonstrated the contamination routes of Y. enterocolitica within the assessed pork production chain, and highlighted the pathogenic potential and antibiotic resistance of this foodborne pathogen.

Virulence-related genes, adhesion and invasion of some Yersinia enterocolitica-like strains suggests its pathogenic potential.

Yersina enterocolitica-like species have not been extensively studied regarding its pathogenic potential. This work aimed to assess the pathogenic potential of some Y. enterocolitica-like strains by evaluating the presence of virulence-related genes by PCR and their ability to adhere to and invade Caco-2 and HEp-2 cells. A total of 50 Y. frederiksenii, 55 Y. intermedia and 13 Y. kristensenii strains were studied. The strains contained the following genes: Y. frederiksenii, fepA(44%), fes(44%) and ystB(18%); Y. intermedia, ail(53%), fepA (35%), fepD(2%), fes(97%), hreP(2%), ystB(2%) and tccC(35%); Y. kristensenii, ail(62%), ystB(23%), fepA(77%), fepD(54%), fes(54%) and hreP(77%). Generally, the Y. enterocolitica-like strains had a reduced ability to adhere to and invade mammalian cells compared to the highly pathogenic Y. enterocolitica 8081. However, Y. kristensenii FCF410 and Y. frederiksenii FCF461 presented high invasion potentials in Caco-2 cells after five days of pre-incubation increased by 45- and 7.2-fold compared to Y. enterocolitica 8081, respectively; but, the ail gene was not detected in these strains. The presence of virulence-related genes in some of the Y. enterocolitica-like strains indicated their possible pathogenic potential. Moreover, the results suggest the existence of alternative virulence mechanisms and that the pathogenicity of Y. kristensenii and Y. frederiksenii may be strain-dependent.

Yersinia enterocolitica YopH-Deficient Strain Activates Neutrophil Recruitment to Peyer's Patches and Promotes Clearance of the Virulent Strain.

Yersinia enterocolitica evades the immune response by injecting Yersinia outer proteins (Yops) into the cytosol of host cells. YopH is a tyrosine phosphatase critical for Yersinia virulence. However, the mucosal immune mechanisms subverted by YopH during in vivo orogastric infection with Y. enterocolitica remain elusive. The results of this study revealed neutrophil recruitment to Peyer's patches (PP) after infection with a YopH-deficient mutant strain (Y. enterocolitica ΔyopH). While the Y. enterocolitica wild-type (WT) strain in PP induced the major neutrophil chemoattractant CXCL1 mRNA and protein levels, infection with the Y. enterocolitica ΔyopH mutant strain exhibited a higher expression of the CXCL1 receptor, CXCR2, in blood neutrophils, leading to efficient neutrophil recruitment to the PP. In contrast, migration of neutrophils into PP was impaired upon infection with Y. enterocolitica WT strain. In vitro infection of blood neutrophils revealed the involvement of YopH in CXCR2 expression. Depletion of neutrophils during Y. enterocolitica ΔyopH infection raised the bacterial load in PP. Moreover, the clearance of WT Y. enterocolitica was improved when an equal mixture of Y. enterocolitica WT and Y. enterocolitica ΔyopH strains was used in infecting the mice. This study indicates that Y. enterocolitica prevents early neutrophil recruitment in the intestine and that the effector protein YopH plays an important role in the immune evasion mechanism. The findings highlight the potential use of the Y. enterocolitica YopH-deficient strain as an oral vaccine carrier.

Whole genome analysis of Yersinia ruckeri isolated over 27 years in Australia and New Zealand reveals geographical endemism over multiple lineages and recent evolution under host selection.

Yersinia ruckeri is a salmonid pathogen with widespread distribution in cool-temperate waters including Australia and New Zealand, two isolated environments with recently developed salmonid farming industries. Phylogenetic comparison of 58 isolates from Australia, New Zealand, USA, Chile, Finland and China based on non-recombinant core genome SNPs revealed multiple deep-branching lineages, with a most recent common ancestor estimated at 18 500 years BP (12 355-24 757 95% HPD) and evidence of Australasian endemism. Evolution within the Tasmanian Atlantic salmon serotype O1b lineage has been slow, with 63 SNPs describing the variance over 27 years. Isolates from the prevailing lineage are poorly/non-motile compared to a lineage pre-vaccination, introduced in 1997, which is highly motile but has not been isolated since from epizootics. A non-motile phenotype has arisen independently in Tasmania compared to Europe and USA through a frameshift in fliI, encoding the ATPase of the flagella cluster. We report for the first time lipopolysaccharide O-antigen serotype O2 isolates in Tasmania. This phenotype results from deletion of the O-antigen cluster and consequent loss of high-molecular-weight O-antigen. This phenomenon has occurred independently on three occasions on three continents (Australasia, North America and Asia) as O2 isolates from the USA, China and Tasmania share the O-antigen deletion but occupy distant lineages. Despite the European and North American origins of the Australasian salmonid stocks, the lineages of Y. ruckeri in Australia and New Zealand are distinct from those of the northern hemisphere, suggesting they are pre-existing ancient strains that have emerged and evolved with the introduction of susceptible hosts following European colonization.

Rapid and efficient differentiation of Yersinia species using high-resolution melting analysis.

The primary goal of clinical microbiology is the accurate identification of the causative agent of the disease. Here, we describe a method for differentiation between Yersinia species using PCR-HRMA. The results revealed species-specific melting profiles. The herein developed assay can be used as an effective method to differentiate Yersinia species.

Phenotypic and genotypic analysis of bio-serotypes of Yersinia enterocolitica from various sources in Brazil.

INTRODUCTION: Yersinia enterocolitica is a well-known foodborne pathogen widely distributed in nature with high public health relevance, especially in Europe. METHODOLOGY: This study aimed to analyze the pathogenic potential of Y. enterocolitica isolated strains from human, animal, food, and environmental sources and from different regions of Brazil by detecting virulence genes inv, ail, ystA, and virF through polymerase chain reaction (PCR), phenotypic tests, and antimicrobial susceptibility analysis. Pulsed-field gel electrophoresis (PFGE) was used for the assessment of phylogenetic diversity. RESULTS: All virulence genes were detected in 11/60 (18%) strains of serotype O:3, biotype 4 isolated from human and animal sources. Ten human strains (4/O:3) presented three chromosomal virulence genes, and nine strains of biotype 1A presented the inv gene. Six (10%) strains were resistant to sulfamethoxazole-trimethoprim, seven (12%) to tetracycline, and one (2%) to amikacin, all of which are used to treat yersiniosis. AMP-CEF-SXT was the predominant resistance profile. PFGE analysis revealed 36 unique pulsotypes, grouped into nine clusters (A to I) with similarity ≥ 85%, generating a diversity discriminatory index of 0.957. Cluster A comprised all bio-serotype 4/O:3 strains isolated from animal and humans sources. CONCLUSIONS: This study shows the existence of strains with the same genotypic profiles, bearing all virulence genes, from human and animal sources, circulating among several Brazilian states. This supports the hypothesis that swine is likely to serve as a main element in Y. enterocolitica transmission to humans in Brazil, and it could become a potential threat to public health as in Europe.

Genotyping of Yersinia enterocolitica biotype 1A strains from clinical and nonclinical origins by pulsed-field gel electrophoresis.

Yersinia enterocolitica biotype 1A (B1A) strains are considered mainly nonpathogenic. However, some studies considered strains of this biotype to be the causal agents of infections in humans and animals. In South America, there are no studies that have compared clinical and nonclinical strains of B1A typed by pulsed-field gel electrophoresis (PFGE) and none that have compared the capability of different enzymes on typing these strains. This study typed 51 Y. enterocolitica B1A strains isolated in Brazil and Chile by PFGE, testing the enzymes XbaI, NotI, and XhoI. The resulting dendrograms discriminated the strains in 47, 40, and 49 pulsotypes generated by the cleavage with the enzymes XbaI, NotI, and XhoI, respectively. The majority of the strains were grouped independently of their clinical or nonclinical origins. The high discriminatory power of PFGE confirmed the heterogeneity of B1A strains but could not divide the strains studied into clusters that differed in the frequency of some virulence genes as observed in studies using other methodologies.

TNFRp55 controls regulatory T cell responses in Yersinia-induced reactive arthritis.

In addition to its well-known pro-inflammatory effects, tumor necrosis factor (TNF) displays anti-inflammatory activities through mechanisms poorly understood. Previously, we reported the development of severe chronic Yersinia enterocolitica-induced reactive arthritis (ReA) in mice lacking the TNF receptor (TNFR)p55. As regulatory T (T(reg)) cells limit chronic inflammation, here we aim to investigate the expansion and function of CD4(+)CD25(+)FoxP3(+) T(reg) cells in the ReA animal model. The number of T(reg) cells as well as the FoxP3 mRNA expression and interleukin (IL)-10 levels were significantly decreased in joint regional lymph nodes (RLNs) of TNFRp55(-/-) mice vs wild-type (WT) mice at the arthritis onset. However, at chronic phase of arthritis, the number of T(reg) cell in TNFRp55(-/-) was similar to WT mice. To explore the in vivo function of T(reg) cells at this chronic phase in WT and TNFRp55-deficient mice, we adoptively transferred CD4(+) T cells from TNFRp55-deficient mice of day 21, into naïve WT or TNFRp55(-/-) mice. When knockout mice were used as recipients we observed higher delayed-type hypersensitivity (DTH) responses and joint inflammation after heat-killed Yersinia (HKY) stimulation. Accordingly, we found higher levels of IL-17, interferon (IFN)-γ, IL-6, transforming growth factor (TGF)-ß1 and IL-12/23p40 and lower IL-10 levels in RLN of paws challenged with HKY in TNFRp55(-/-) recipient mice. In addition, we found that CD4(+) T cells from TNFRp55(-/-) mice controlled antigen-specific IL-12/23(p40) production in recipient WT mice. Our results show that TNFRp55 controls the induction and function of T(reg) cells through differential regulation of cytokine production, suggesting a novel molecular target for immune intervention in ReA.

Yersiniose em Trachinotus marginatus (pampo): diagnóstico histopatológico e imuno-histoquímico / Yersiniosis in Trachinotus marginatus (pampo): histopathological and immunohistochemical diagnostic

Descreveu-se um surto de yersiniose em uma criação de pampo Trachinotus marginatus estudado no Laboratório de Piscicultura Estuarina e Marinha da Universidade Federal do Rio Grande. A enfermidade manifestou-se por transtornos natatórios, exoftalmia com panoftalmite e pela clássica boca vermelha, nome pelo qual se denomina "enfermidade da boca vermelha" (EBV). Na necropsia, foram observados focos de hemorragia peritoneal, esplenomegalia e hepatomegalia. Microscopicamente, foi observada panoftalmite com infiltrados inflamatórios densos que afetavam quase todas as estruturas oculares. Esses infiltrados estavam constituídos por granulócitos, linfócitos, macrófagos e células granulares eosinofílicas. No exsudado, observaram-se estruturas pequenas, pouco coradas, de aspecto bacteriano Gram negativo. O estudo imuno-histoquímico, que se utilizou de um anticorpo monoclonal anti-Yersinia ruckeri, resultou positivo. Este é o primeiro surto conhecido de yersiniose em Trachinotus marginatus no Brasil.

In the "Laboratório de Piscicultura Estuarina e Marinha da Universidade Federal do Rio Grande" the rearing of Trachinotus marginatus (pompano) is studied, and in one of these rearing a yersioniosis outbreak occured. The disease was manifested by swimming disturbances, exophthalmiawith panophthalmitis and the classic red mouth, (red mouth disease (RMD). The necropsies revealed focus of peritoneal hemorrhage, esplenomegalyand hepatomegaly.Microscopically a panophthalmitis with dense inflammatory infiltrates was observed, which affected almost all ocular structures. These infiltrates were constituted by granulocytes, lymphocytes, macrophages and eosinophylicgranular cells (ECG), in the exudatessmall structures less colored with Gram negative bacteria aspect were observed. The immunohistochemical, which used a monoclonal antibody anti- Yersinia ruckeri waspositive. The RMD is caused by enterobacteriaGram negative, which affects both freshwater and saltwater fish, with predominance in salmonids. This is the first known yersiniosis outbreak in Trachinotus marginatus in Brazil and it is necessary to keep this disease in mind at the moment of rearing fish of this species.

A polyphasic approach to study the intraspecific diversity of Yersinia ruckeri strains isolated from recent outbreaks in salmonid culture.

A polyphasic analysis was carried out on Yersinia ruckeri strains isolated from recently outbreaks in vaccinated fish using a combination of different phenotypic and molecular typing methods in order to study their variability and epidemiological relationships. Eighty strains were subjected to biotyping with conventional tests and API 20E system, serotyping, outer membrane protein (OMP) and lipopolysaccharide (LPS) profiling, and genetic fingerprinting by ERIC-PCR and REP-PCR techniques. The strains showed a high diversity, as evidenced by the formation of different phenotypic groups mainly related to the serotypes, LPS and OMP profiles. The diversity among all isolates, calculated as Simpson's diversity index (Di), varied between 0.35 (REP-PCR) and 0.70 (OMP). The most discriminative values (Di value ≥0.86) were obtained from any combination of three methods including biotype, serotype, API 20E profile, LPS or OMP. With the combination of all typing methods used a Di value of 0.90 was obtained. Association between different groups to the host species was evidenced. Furthermore, it seems that strains with similar characteristics are associated with recent outbreaks occurred in vaccinated fish in certain geographical areas. Our results emphasize the usefulness of using a combination of several different typing methods for epidemiological and bacterial diversity studies.

Yersiniose em Trachinotus marginatus (pampo): diagnóstico histopatológico e imuno-histoquímico / Yersiniosis in Trachinotus marginatus (pampo): histopathological and immunohistochemical diagnostic

Descreveu-se um surto de yersiniose em uma criação de pampo Trachinotus marginatus estudado no Laboratório de Piscicultura Estuarina e Marinha da Universidade Federal do Rio Grande. A enfermidade manifestou-se por transtornos natatórios, exoftalmia com panoftalmite e pela clássica boca vermelha, nome pelo qual se denomina "enfermidade da boca vermelha" (EBV). Na necropsia, foram observados focos de hemorragia peritoneal, esplenomegalia e hepatomegalia. Microscopicamente, foi observada panoftalmite com infiltrados inflamatórios densos que afetavam quase todas as estruturas oculares. Esses infiltrados estavam constituídos por granulócitos, linfócitos, macrófagos e células granulares eosinofílicas. No exsudado, observaram-se estruturas pequenas, pouco coradas, de aspecto bacteriano Gram negativo. O estudo imuno-histoquímico, que se utilizou de um anticorpo monoclonal anti-Yersinia ruckeri, resultou positivo. Este é o primeiro surto conhecido de yersiniose em Trachinotus marginatus no Brasil.(AU)

In the "Laboratório de Piscicultura Estuarina e Marinha da Universidade Federal do Rio Grande" the rearing of Trachinotus marginatus (pompano) is studied, and in one of these rearing a yersioniosis outbreak occured. The disease was manifested by swimming disturbances, exophthalmiawith panophthalmitis and the classic red mouth, (red mouth disease (RMD). The necropsies revealed focus of peritoneal hemorrhage, esplenomegalyand hepatomegaly.Microscopically a panophthalmitis with dense inflammatory infiltrates was observed, which affected almost all ocular structures. These infiltrates were constituted by granulocytes, lymphocytes, macrophages and eosinophylicgranular cells (ECG), in the exudatessmall structures less colored with Gram negative bacteria aspect were observed. The immunohistochemical, which used a monoclonal antibody anti- Yersinia ruckeri waspositive. The RMD is caused by enterobacteriaGram negative, which affects both freshwater and saltwater fish, with predominance in salmonids. This is the first known yersiniosis outbreak in Trachinotus marginatus in Brazil and it is necessary to keep this disease in mind at the moment of rearing fish of this species.(AU)