Genetic Analysis and Serological Detection of Novel O-Antigen Gene Clusters of Plesiomonas shigelloides.

Plesiomonas shigelloides, a member of the family Vibrionaceae, is a gram-negative, rod-shaped, facultative anaerobic bacterium with flagella. P. shigelloides has been isolated from such sources as freshwater, surface water, and many wild and domestic animals. P. shigelloides contains 102 Oantigens and 51 H-antigens. The diversity of O-antigen gene clusters is relatively poorly understood. In addition to O1 and O17 reported by other laboratories, and the 12 O serogroups (O2, O10, O12, O23, O25, O26, O32, O33, O34, O66, O75, and O76) reported previously by us, in the present study, nine new P. shigelloides serogroups (O8, O17, O18, O37, O38, O39, O44, O45, and O61) were sequenced and annotated. The genes for the O-antigens of these nine groups are clustered together in the chromosome between rep and aqpZ. Only O38 possesses the wzm and wzt genes for the synthesis and translocation of O-antigens via the ATP-binding cassette (ABC) transporter pathway; the other eight use the Wzx/Wzy pathway. Phylogenetic analysis using wzx and wzy showed that both genes are diversified. Among the nine new P. shigelloides serogroups, eight use wzx/wzy genes as targets. In addition, we developed an O-antigen-specific PCR assay to detect these nine distinct serogroups with no cross reactions among them.

Aeromonas and Plesiomonas species from scarlet ibis (Eudocimus ruber) and their environment: monitoring antimicrobial susceptibility and virulence.

The present study aimed at evaluating the role of captive scarlet ibises (Eudocimus ruber) and their environment as reservoirs of Aeromonas spp. and Plesiomonas spp., and analyzing the in vitro antimicrobial susceptibility and virulence of the recovered bacterial isolates. Thus, non-lactose and weak-lactose fermenting, oxidase positive Gram-negative bacilli were recovered from cloacal samples (n = 30) of scarlet ibises kept in a conservational facility and from water samples (n = 30) from their environment. Then, the antimicrobial susceptibility, hemolytic activity and biofilm production of the recovered Aeromonas spp. and Plesiomonas shigelloides strains were assessed. In addition, the virulence-associated genes of Aeromonas spp. were detected. Ten Aeromonas veronii bv. sobria, 2 Aeromonas hydrophila complex and 10 P. shigelloides were recovered. Intermediate susceptibility to piperacillin-tazobactam and cefepime was observed in 2 Aeromonas spp. and 1 P. shigelloides, respectively, and resistance to gentamicin was observed in 4 P. shigelloides. The automated susceptibility analysis revealed resistance to piperacillin-tazobactam and meropenem among Aeromonas spp. and intermediate susceptibility to gentamicin among P. shigelloides. All Aeromonas isolates presented hemolytic activity, while 3 P. shigelloides were non-hemolytic. All Aeromonas spp. and 3/10 P. shigelloides were biofilm-producers, at 28 °C, while 10 Aeromonas spp. and 6/10 P. shigelloides produced biofilms, at 37 °C. The most prevalent virulence genes of Aeromonas spp. were asa1 and ascV. Scarlet ibises and their environment harbour potentially pathogenic bacteria, thus requiring monitoring and measures to prevent contamination of humans and other animals.

Plesiomonas shigelloides Revisited.

After many years in the family Vibrionaceae, the genus Plesiomonas, represented by a single species, P. shigelloides, currently resides in the family Enterobacteriaceae, although its most appropriate phylogenetic position may yet to be determined. Common environmental reservoirs for plesiomonads include freshwater ecosystems and estuaries and inhabitants of these aquatic environs. Long suspected as being an etiologic agent of bacterial gastroenteritis, convincing evidence supporting this conclusion has accumulated over the past 2 decades in the form of a series of foodborne outbreaks solely or partially attributable to P. shigelloides. The prevalence of P. shigelloides enteritis varies considerably, with higher rates reported from Southeast Asia and Africa and lower numbers from North America and Europe. Reasons for these differences may include hygiene conditions, dietary habits, regional occupations, or other unknown factors. Other human illnesses caused by P. shigelloides include septicemia and central nervous system disease, eye infections, and a variety of miscellaneous ailments. For years, recognizable virulence factors potentially associated with P. shigelloides pathogenicity were lacking; however, several good candidates now have been reported, including a cytotoxic hemolysin, iron acquisition systems, and lipopolysaccharide. While P. shigelloides is easy to identify biochemically, it is often overlooked in stool samples due to its smaller colony size or relatively low prevalence in gastrointestinal samples. However, one FDA-approved PCR-based culture-independent diagnostic test system to detect multiple enteropathogens (FilmArray) includes P. shigelloides on its panel. Plesiomonads produce ß-lactamases but are typically susceptible to many first-line antimicrobial agents, including quinolones and carbapenems.

PREVALENCE, BIOCHEMICAL CHARACTERISTICS, AND ANTIBIOTIC SUSCEPTIBILITY OF AEROMONADS, VIBRIOS, AND PLESIOMONADS ISOLATED FROM DIFFERENT SOURCES AT A ZOO.

Aeromonas spp., Vibrio parahaemolyticus , and Plesiomonas shigelloides are commonly implicated in foodborne and waterborne diarrheal illnesses of humans and other animals. The present study assessed the prevalence, biochemical characteristics, and antibiotic susceptibility of Aeromonas spp., V. parahaemolyticus , and P. shigelloides by analyzing samples from 729 sources at a zoo, including animal feces (n=607), watering facilities (n=104), and pond water samples (n=18). Of the 729 samples collected, 40 (5.5%) contained one of these four species of bacteria: A. hydrophila (n=16; 2.2%), A. sobria (n=12; 1.6%), V. parahaemolyticus (n=10; 1.4%), and P. shigelloides (n=2; 0.3%). The 16 isolates of A. hydrophila came from three fecal samples, eight watering facilities, and five pond water samples. The 12 isolates of A. sobria came from four fecal samples, three watering facilities, and five pond water samples. The 10 isolates of V. parahaemolyticus came from one fecal sample and nine watering facilities. The two isolates of P. shigelloides came from one watering facility and one pond water sample. Of the 40 isolates, 16 (40.0%), 21 (52.5%), and three (7.5%) originated from mammals, birds, and reptiles, respectively. All isolates tested positive for NO3, tryptophan, p-nitrophenyl-ß-D-galactopyranoside, glucose assimilation, N-acetyl-glucosamine, maltose, gluconate, malate, and oxidase. Aeromonas spp. and V. parahaemolyticus exhibited similar biochemical characteristics, whereas P. shigelloides exhibited distinct fermentation characteristics. All the isolated strains exhibited hemolytic activity; variable results of DNase, protease, and Congo red uptake tests; and resistance to ampicillin, bacitracin, novobiocin, penicillin, and vancomycin. All the strains were sensitive to amikacin, chloramphenicol, colistin, gentamicin, kanamycin, norfloxacin, and trimethoprim-sulfadimethoxazole. Because of the high proportion of asymptomatic carriers of these potentially pathogenic bacteria and their wide distribution, consistent monitoring of food and water sources is necessary to prevent disease outbreaks.

[Identification and pathological observation of a pathogenic Plesiomonas shigelloides strain isolated from cultured tilapia (Oreochromis niloticus)].

OBJECTIVE: A mass mortality of tilapia broke out in an aquaculture farm in Panyu, Guangdong Province in May, 2013. Affected fish showed blackening of body color, haemorrhageing on surface, scales shedding, fin rotting, and the presence of yellow liver, dark red spleen, enlarged gallbladder and ascitic fluid in the abdominal cavity. The purpose of this research was isolating and identifying the pathogen. METHODS: We isolated a suspicious bacteria strain PYS1 from diseased fish with significant pathological signs. The homology of 16S rRNA gene sequence of strain PYS1 and its morphological, cultural, and physical and chemical characteristics were studied for its identification. Its pathogenicity was investigated by recursive infection experiment and histopathological study. Its effective medicines was screened by antibiotic sensitive test. RESULTS: The results showed that strain PYS1 was Plesiomonas shigelloides clustered with P. shigelloides strains isolated from other fishes in the molecular phylogenetic tree of 16S rRNA gene sequences. Strain PYS1 was multiple drug resistant and only sensitive to a small part of 31 tested antibiotics (e.g., ceftriaxone, cefaclor, cefazolin, etc.). The symptoms of tilapia (O. niloticus) artificially infected with strain PYS1 were similar with natural infected fish. The half lethal dose (LD50) of strain PYS1 to tilapia was 1.425 x 10(8) CFU per fish. Paraffin sections showed intestine, liver, spleen, kidney and heart tissue injury caused by the strain. CONCLUSION: Our study demonstrated that P. shigelloides was the pathogen of cultured tilapia in the aquaculture farm and first reported its bacterial pathogenicity on Nile tilapia.

[Identification and drug sensitivity of a Plesiomonas shigelloides isolated from diseased sturgeons].

OBJECTIVE: In 2012, the cumulative mortality of farmed sturgeons in Beijing was almost 60% with various symptoms, including the reddening of the anus with yellow exudation, ascities in the peritoneal cavity, petechial haemorrhages in liver and internal muscle wall, and the swollen spleen. METHODS: We isolated the pathogen from the dying sturgeons with significant pathological signs, and then analyzed its morphological, physiological and biochemical characteristics, taxonomic status, and drug sensitivity. Moreover, the pathogenic characteristic of presumptive pathogens was identified by artificial infection. RESULTS: The 16S rDNA sequence of the pathogen was more than 99% homology with that of Plesiomonas shigelloides, suggesting that the pathogen was P. shigelloides, which was also demonstrated by the results of biochemical tests. The LD50 of the pathogen to sturgeon was 1.0 x 10(5.8) CFU/mL, and it also can cause liver, kidey and spleen to lesions. There were no activities of amylase, caseinase, lipase, gelatinase and haemolysis of extracellular products of P. shigelloides, and its toxicity might be from endotoxin. In addition, the bacterium was specific sensitive to enrofloxacin, doxycyline hyclate, florfenicol and thiamphenicol with MIC less than 2 microg/mL. CONCLUSION: P. shigelloides was the main pathogen to cultured sturgeons in Beijing area, and enrofloxacin, doxycyline hyclate and florfenicol can be used against the disease.

Core oligosaccharide of Plesiomonas shigelloides PCM 2231 (Serotype O17) lipopolysaccharide--structural and serological analysis.

The herein presented complete structure of the core oligosaccharide of lipopolysaccharide (LPS) P. shigelloides Polish Collection of Microorganisms (PCM) 2231 (serotype O17) was investigated by (1)H, (13)C NMR spectroscopy, mass spectrometry, chemical analyses and serological methods. The core oligosaccharide is composed of an undecasaccharide, which represents the second core type identified for P. shigelloides serotype O17 LPS. This structure is similar to that of the core oligosaccharide of P. shigelloides strains 302-73 (serotype O1) and 7-63 (serotype O17) and differs from these only by one sugar residue. Serological screening of 55 strains of P. shigelloides with the use of serum against identified core oligosaccharide conjugated with bovine serum albumin (BSA) indicated the presence of similar structures in the LPS core region of 28 O-serotypes. This observation suggests that the core oligosaccharide structure present in strain PCM 2231 could be the most common type among P. shigelloides lipopolysaccharides.

Molecular evidence of Plesiomonas shigelloides as a possible zoonotic agent.

The most frequently used method for establishing epidemiological relationships between Plesiomonas shigelloides strains is O:H serotyping. However, a number of strains are not serotypeable and isolates from diverse sources can display the same serovar. Moreover, since the zoonotic nature of Plesiomonas has been suggested and this hypothesis is based on the identical serovars found in animals and humans, we intend to use four DNA-based techniques: random amplified polymorphic DNA-PCR, enterobacterial repetitive intergenic consensus-PCR, repetitive extragenic palindromic-PCR, and pulsed field gel electrophoresis in order to screen 24 strains belonging to nine O:H serovars isolated from humans, animals, and the environment. In general, P. shigelloides showed a high genetic heterogeneity. Three pairs of strains, each containing a human and an animal isolate, displayed similar genotypes. This is the first report that provides molecular evidence that P. shigelloides may be zoonotic.

Identification of Plesiomonas spp.: serological and MALDI-TOF MS methods.

Biochemical and serological profiles of isolates of Plesiomonas shigelloides were assayed using standard procedures in isolates from various clinical samples. Seventy-four isolates, including P. shigelloides type strain, were further characterized by MALDI-TOF MS using 3-methoxy-4-hydroxycinnamic acid as matrix. Multiple ions in the 3- to 12-kDa mass range were found in the spectra of each strain, from which the "species-identifying" unique biomarker ions were identified. After creating the species-specific patterns, a spectral database was generated for reliable, rapid, reproducible and accurate identification of Plesiomonas strains. The classical strain description (biochemical and serological) was thus complemented with the metabolic (proteomic) characterization.

Recombining population structure of Plesiomonas shigelloides (Enterobacteriaceae) revealed by multilocus sequence typing.

Plesiomonas shigelloides is an emerging pathogen that is widespread in the aquatic environment and is responsible for intestinal diseases and extraintestinal infections in humans and other animals. Virtually nothing is known about its genetic diversity, population structure, and evolution, which severely limits epidemiological control. We addressed these questions by developing a multilocus sequence typing (MLST) system based on five genes (fusA, leuS, pyrG, recG, and rpoB) and analyzing 77 epidemiologically unrelated strains from several countries and several ecological sources. The phylogenetic position of P. shigelloides within family Enterobacteriaceae was precisely defined by phylogenetic analysis of the same gene portions in other family members. Within P. shigelloides, high levels of nucleotide diversity (average percentage of nucleotide differences between strains, 1.49%) and genotypic diversity (64 distinct sequence types; Simpson's index, 99.7%) were found, with no salient internal phylogenetic structure. We estimated that homologous recombination in housekeeping genes affects P. shigelloides alleles and nucleotides 7 and 77 times more frequently than mutation, respectively. These ratios are similar to those observed in the naturally transformable species Streptococcus pneumoniae with a high rate of recombination. In contrast, recombination within Salmonella enterica, Escherichia coli, and Yersinia enterocolitica was much less frequent. P. shigelloides thus stands out among members of the Enterobacteriaceae. Its high rate of recombination results in a lack of association between genomic background and O and H antigenic factors, as observed for the 51 serotypes found in our sample. Given its robustness and discriminatory power, we recommend MLST as a reference method for population biology studies and epidemiological tracking of P. shigelloides strains.

[Isolation and identification of species from the genera Aeromonas, Vibrio, and Plesiomonas from extraintestinal samples in Cuba]. / Aislamiento e identificación de especies pertenecientes a los géneros Aeromonas, Vibrio y Plesiomonas procedentes de muestras extra-intestinales en Cuba.

Extraintestinal infections caused by the genera Aeromonas, Vibrio and Plesiomonas have high morbidity and mortality rates in different areas of world. From January 2002 to December 2003, the National Reference Laboratory for Acute Diarrhoeal Diseases of the Pedro Kourí Tropical Medicine Institute received 95 gram-negative, facultative anaerobic, oxidase positive bacilli strains from different extraintestinal specimen (blood, ear exudates, infected wounds, conjunctive exudates, urine, and catheters, among others) sent by different provincial laboratories along the country. Aeromonas caviae, Aeromonas veronii bv sobria, Aeromonas jandaei, Vibrio cholerae no-O1, Vibrio vulnificus, Vibrio fluvialis and Plesiomonas shigelloides were the species most frequently found in the sample analysed.

Aislamiento e identificación de especies pertenecientes a los géneros Aeromonas, Vibrio y Plesiomonas procedentes de muestras extra-intestinales en Cuba / Isolation and identification of species from the genera Aeromonas, Vibrio, and Plesiomonas from extraintestinal samples in Cuba

Las infecciones extra-intestinales producidas por los géneros Aeromonas, Vibrio y Plesiomonas presentan una elevada tasa de morbimortalidad en diferentes áreas geográficas. De enero 2002 a diciembre 2003 se recibieron en el Laboratorio Nacional de Referencia de Enfermedades Diarreicas Agudas del Instituto de Medicina Tropical Pedro Kourí, 95 cepas de bacilos gramnegativos, anaerobios facultativos, oxi-dasa positiva, procedentes de muestras extra-intestinales (hemocultivos, exudados óticos, pus de heridas, exudados conjuntivales, urocultivos, catéter, entre otras) remitidas de diferentes provincias del país. Se demostró la presencia de Aeromonas caviae, Aero-monas veronii bv sobria, Aeromonas jandaei, Vibrio cholerae no -O1, Vibrio vulnificus, Vibrio fluvialis y Plesiomonas shigelloides en las muestras estudiadas.

Extraintestinal infections caused by the genera Aeromonas, Vibrio and Plesiomonas have high morbidity and mortality rates in different areas of world. From January 2002 to December 2003, the National Reference Laboratory for Acute Diarrhoeal Diseases of the Pedro Kourí Tropical Medicine Institute received 95 gramnegative, facultative anaerobic, oxidase positive bacilli strains from different extraintestinal specimen (blood, ear exudates, infected wounds, conjunctive exudates, urine, and catheters, among others) sent by different provincial laboratories along the country. Aeromonas caviae, Aeromonas veronii bv sobria, Aeromonas jandaei, Vibrio cholerae no-O1, Vibrio vulnificus, Vibrio fluvialis and Plesiomonas shigelloides were the species most frequently found in the sample analysed.

Differentiation of Aeromonas isolated from drinking water distribution systems using matrix-assisted laser desorption/ionization-mass spectrometry.

The genus Aeromonas is one of several medically significant genera that have gained prominence due to their evolving taxonomy and controversial role in human diseases. In this study, matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) was used to analyze the whole cells of both reference strains and unknown Aeromonas isolates obtained from water distribution systems. A library of over 45 unique m/z signatures was created from 40 strains that are representative of the 17 recognized species of Aeromonas, as well as 3 reference strains from genus Vibrio and 2 reference strains from Plesiomonas shigelloides. The library was used to help speciate 52 isolates of Aeromonas. The environmental isolates were broken up into 2 blind studies. Group 1 contained isolates that had a recognizable phenotypic profile and group 2 contained isolates that had an atypical phenotypic profile. MALDI-MS analysis of the water isolates in group 1 matched the phenotypic identification in all cases. In group 2, the MALDI-MS-based determination confirmed the identity of 18 of the 27 isolates. These results demonstrate that MALDI-MS analysis can rapidly and accurately classify species of the genus Aeromonas, making it a powerful tool especially suited for environmental monitoring and detection of microbial hazards in drinking water.

[Biological characteristics of Plesiomonas shigelloides strains of human and animal origin]. / Biologické vlastnosti kmenov Plesiomonas shigelloides humánneho a veterinárneho pôvodu.

We focused on serotyping and biological characteristics of Plesiomonas shigelloides strains potentially associated with virulence. Thirteen strains isolated from humans (H) and 14 strains of animal origin (A) were tested. The most frequent serotype among H strains was 040:H6 while 066:H3 predominated among A strains. All of the H strains and 92.8% of A strains were hydrophobic. H isolates showed lower motility (30.1 mm) compared to A isolates (46.8 mm). As many as 76.9% and 71.4% of H and A strains, respectively, produced beta-hemolysis. Both H and A strains exhibited low biofilm production on a glass surface. No significant differences were found between H and A strains in lipase production and histidine decarboxylase production. The zones of bacterial growth inhibition in response to oxidative stress were on average 26.6 mm and 28.1 mm for H and A strains, respectively. None of the strains tested produced unsubstituted short-chain acyl homoserine lactones. Our results showed that tested Plesiomonas shigelloides strains produced multiple potential virulence factors that may play a role in the pathogenesis of infections caused by this agent.

Two cases of continuous ambulatory peritoneal dialysis-associated peritonitis due to Plesiomonas shigelloides.

We describe herein the first two cases of Plesiomonas shigelloides continuous ambulatory peritoneal dialysis-related peritonitis. Both patients presented with abdominal pain and turbid dialysis effluent with or without fever. Both recovered after 10 days of intraperitoneal administration of cefazolin and tobramycin. The route of transmission may have been direct contamination of the connection device or bacterial translocation through the gastrointestinal tract.

Serotypes and anti-microbial susceptibility of Plesiomonas shigelloides isolates from humans, animals and aquatic environments in different countries.

A total of 73 strains of Plesiomonas shigelloides isolated from humans (24 strains) animals (21 strains) and aquatic environment (28 strains) were determined for their O:H serotype and susceptibility to 18 anti-microbial substances and to the vibriostatic agent O/129. Of all strains, 86.3% were typeable by the O and 94.5% by the H anti-sera used. The serotype distribution was heterogeneous within a country and between the countries. Of the 57 different serotypes identified, O11:H2 (2 strains), O22:H3 (4 strains), O35:HH11 (2 strains), O52:H3 (2 strains) and O90:H6 (2 strains) were found among isolates from humans and animals (mainly in cats) in Finland and Cuba, and O23:H1a1b (3 strains) among isolates from environmental sources in Slovak Republic and Italy. Most (93-100%) of all strains were susceptible to all anti-microbials tested but resistant (92-96%) to the broad-spectrum penicillins (ampicillin, mezlocillin). No correlation between anti-microbial resistance patterns and serotypes was found.

Potential virulence-associated properties of Plesiomonas shigelloides strains.

Serotyping and some potential virulence-associated markers were investigated in Plesiomonas shigelloides strains isolated from humans, animals and aquatic environments. Surface properties of these strains were evaluated using Congo red binding, salt-aggregation test, bacterial adherence to xylene and motility. Production of pancreatic elastase, proteinase (consistent with subtilisin Carlsberg), triacylglycerol lipase, histidine decarboxylase and beta-hemolysin was also determined. In addition, detection of signal molecules such as C4-C8 unsubstituted N-acylhomoserine lactones (AHLs) was performed. The serological typing of the P. shigelloides strains showed that the isolates belonged to 13 different serovars. The majority of the strains were hydrophobic and motile. The strains produced low levels of elastase, proteinase and histidine decarboxylase whereas triacylglycerol lipase activity was relatively high. Only 23.3 % of the strains produced hemolysin. The AHLs signal molecules were not detected. P. shigelloides strains were able to produce a variety of potential virulence markers which may be involved in the pathogenesis of Plesiomonas-associated infections.

Occurrence, characterisation and detection of potential virulence determinants of emerging aquatic bacterial pathogens from the Philippines and Thailand.

Strains of Aeromonas spp., 'non-cholera vibrios' (NCVs) and Plesiomonas shigelloides isolated from aquatic environments, fish and human diarrhoeal cases in the Philippines and Thailand were characterised for potential virulence markers. Thus, the production of cytotoxin, cell-associated and cell-free haemolysin and their capacity to adhere to human intestinal (Henle 407) cells in vitro was investigated. In addition, the occurrence of tlh and tdh haemolysin genes and urease activity among V. parahaemolyticus strains was investigated. The results showed that strains recovered from clinical sources (human and fish) produced these virulence factors, whereas these are absent in environmental strains.

beta-Lactam-susceptibility patterns of Plesiomonas shigelloides strains: importance of inoculum and medium.

The susceptibility of 10 Plesiomonas shigelloides strains to 30 beta-lactam antibiotics was examined. Susceptibility testing was carried out with a microdilution procedure using 3 media (Isosensitest broth, Mueller-Hinton broth and cation-adjusted Mueller-Hinton broth) and 4 inocula [1 x 10(4), 1 x 10(5), 1 x 10(6) and 1 x 10(7) colony-forming units (CFU)/ml]. A high inoculum dependency of MIC values was found for numerous beta-lactams. Each strain showed 2 completely different susceptibility patterns to several cephalosporins and aztreonam, but the same patterns were found with little variation in each strain. Using an inoculum of 1 x 10(4) CFU/ml the strains showed a high susceptibility to aztreonam and to all cephalosporins in all media (pattern 1), whereas they showed a decreased susceptibility to aztreonam and numerous cephalosporins with an inoculum of 1 x 10(7) CFU/ml (pattern 2). Using an inoculum of 1 x 10(6) CFU/ml, 4/10 strains in lsosensitest broth and 2/10 strains in Mueller-Hinton media showed pattern 1. Susceptibility testing of numerous penicillins revealed a medium-independent inoculum dependency, characterized by a step-to-step correlation between MICs and inocula. The beta-lactam susceptibility patterns arising from different inocula point to new beta-lactamase expression and regulation mechanisms in Plesiomonas. The potential to be resistant to a variety of beta-lactams under conceivable testing conditions should question the use of numerous beta-lactams for the treatment of Plesiomonas infections.

[Plesiomonas shigelloides, a Vibrionaceae to be taken into account]. / Plesiomonas shigelloides una Vibrionaceae en quien pensar.

The antigenic structure and antimicrobial susceptibility were studied in 99 strains isolated from patients with acute diarrhea (6 strains from an outbreak of digestive transmission disease in Santiago de Cuba) and a strain isolated from a patient who died from infections neurological syndrome (INS, meningitis). Four new serotypes (093, 994, 095, 096), which had not been described in the world classification, were identified from the Cuban isolated strains and were included in the International Serotyping Scheme by the International Reference Center located in Prague, Czech Republic. For the first time in Cuba, the circulation of serotypes 017:H11, 011: H2, 023. H1alc, 057: H3 which show cross reaction to Shiguella species was proved. Those strains from the outbreak of digestive disease belonged to serotype 050: H11 and had a thermostable toxin. The first case of infectious neurologic syndrome with Plesionomas shigelloides etiology reported in Cuba was described; the strain corresponded to serotype 050: H11. The worldwide reported pattern of antimicrobial resistance was demonstrated.