Enfermedad neonatal temprana por Streptococcus Agalactiae en un unidad de recién nacidos, factores de riesgo maternofetales asocaidos a severidad y mortalidad

la colonización materna por estreptococo del grupo B (SGB) en países en desarrollo es de 4-20%, 50% de sus hijos nacen colonizados y el 1-2% desarrollan enfermedad invasiva con alto riesgo de morbimortalidad y secuelas. La incidencia de infección es diez veces más alta en menores de 1500gramos. Objetivo: determinar los factores de riesgo maternofetales asociados a enfermedad severa y mortalidad neonatal por Streptococcus agalactidae en una unidad de recién nacidos. Materiales y Métodos: se realizó un estudio observacional analítico de cohorte histórica durante un periodo de 2 años. Se tomaron 11 (once) recién nacidos con cuadro clínico de enfermedad invasiva por SGB, con confirmación en ocho (8) casos con hemocultivos, un (1) caso con cultivo de líquido cefalorraquídeo y dos (2) con ambos. Quince (15) controles que correspondían a las madres colonizadas con recién nacidos asintomáticos y con cultivos negativos. Las características demográficas de los dos (2) grupos no mostraron diferencias significativas. Se calcularon frecuencias absolutas y relativas y se buscaron asociaciones mediante el cálculo del estadístico Chi2, se aceptaron valores de p < 0.05, bajo el programa SPSS 15.0 para Windows. Resultados: los factores maternos predictivos para enfermedad por SGB incluyeron, fiebre periparto mayor a 37.5 grados centígrados (p <0.05), corioamnionitis y ruptura de membranas mayor a 18 horas (p<0.05). Los factores de riesgo neonatal incluyeron prematurez (<37 semanas) y bajo peso al nacer(<2500 gramos) (p<0.05). La severidad de la enfermedad se valoró por la presencia de neumonía, meningitis o hemorragia pulmonar. Se encontró una mortalidad de 5(45%). Conclusiones: se encontró relación estadísticamente significativa entre la corioaminionitis materna, la ruptura de membranas mayor a 18 horas, la prematurez y el peso bajo al nacer con la severidad de la enfermedad y la mortalidad. La incidencia estimada de infección neonatal en la unidad de recién nacidos fue de 1,8 casos/1000 nacidos vivos y la de colonización materna fue de 4,3 casos/ 1000 maternas. Se deben realizar más estudios en el país para establecer la incidencia real de enfermedad neonatal por SGB y hacer investigación sobre la costoefectividad de las medidas de prevención.

Enfermedad neonatal temprana por Streptococcus agalactiae en una unidad de recién nacidos, factores de riesgo materno-fetales asociados a severidad y mortalidad

La colonización materna por estreptococo del grupo B (SGB) en países en desarrollo es del 4-20%; el 50% de sus hijos nacen colonizados y el 1-2% desarrollan enfermedad invasiva con alto riesgo de morbimortalidad y secuelas. Objetivo: determinar los factores de riesgo materno-fetales asociados a enfermedad severa y mortalidad neonatal por SGB en una unidad de recién nacidos. Materiales y métodos: se realizó un estudio observacional analítico de cohorte histórica durante un período de dos años. Se tomaron 11 casos con enfermedad invasiva y 15 controles. Se calcularon frecuencias absolutas y relativas, y se buscaron asociaciones mediante el cálculo del estadístico chi2. Resultados: los factores maternos predictivos para enfermedad por SGB incluyeron fiebre periparto mayor a 37,5 grados centígrados (p < 0,05), corioamnionitis y ruptura de membranas mayor a 18 horas (p < 0,05). Los factores de riesgo neonatal incluyeron prematurez (< 37 semanas) y bajo peso al nacer (< 2.500 gramos) (p < 0,05). Se encontró una mortalidad de 5 (45%). Conclusiones: hubo relación estadísticamente significativa entre la corioaminionitis materna, la ruptura de membranas mayor a 18 horas, la prematurez y el peso bajo al nacer con la severidad de la enfermedad y la mortalidad. La incidencia estimada de infección neonatal en la unidad de recién nacidos fue de 1,8 casos/1.000 nacidos vivos, y la de colonización materna fue de 4,3 casos/1.000 maternas. Se deben realizar más estudios en el país para establecer la incidencia real de enfermedad neonatal por SGB y hacer investigación sobre la costo-efectividad de las medidas de prevención.

Neonatal mortality in puppies due to bacteremia by Streptococcus dysgalactiae subsp dysgalactiae

We report a case of bacteremia in puppies caused by Streptococcus dysgalactiae subsp. dysgalactiae. Identification was achieved by phenotypic and molecular genetic methods. This is the first report of the recovery of S. dysgalactiae subsp. dysgalactiae from dogs.

Streptococcus castoreus sp nov., isolated from a beaver (Castor fiber)

A previously undescribed, Gram-positive, catalase-negative, Streptococcus-like organism originating from a European beaver (Castor fiber) was subjected to a taxonomic study. The organism displayed beta-haemolytic activity and gave a positive reaction with Lancefield group A antisera. Based on the results of biochemical testing, the organism was tentatively identified as a member of the genus Streptococcus, but it did not correspond phenotypically to any recognized species of this genus. Comparative 16S rRNA gene sequencing studies confirmed this assignment, with the bacterium forming a hitherto unknown subline within the genus. Sequence divergence values of greater than 3% from other reference streptococcal species, however, demonstrated that the unidentified coccus-shaped organism represents a hitherto unknown species. Based on phenotypic and molecular phylogenetic evidence, it is therefore proposed that the unknown organism from a beaver be classified as a novel species, Streptococcus castoreus sp. nov. The type strain is M605815/03/2(T) (=CCUG 48115(T) = CIP 108205(T)).

Streptococcus halichoeri sp nov., isolated from grey seals (Halichoerus grypus)

Phenotypic and phylogenetic studies were performed on six unidentified, Gram-positive, catalase-negative, chain-forming Streptococcus-like organisms recovered from grey seals. Biochemically the six strains were highly related to each other, but they did not appear to correspond to any recognized species of the genus Streptococcus. Comparative 16S rRNA gene sequencing studies confirmed that phylogenetically the strains were members of the genus Streptococcus, but sequence divergence values of greater than 3 % compared with reference streptococcal species demonstrated that the organisms from seals represent a novel species. SDS-PAGE analysis of whole-cell proteins confirmed the phenotypic distinctiveness of the seal organisms. Based on biochemical criteria and molecular chemical and genetic evidence, it is proposed that the unknown organism from seals be classified as a novel species, Streptococcus halichoeri sp. nov., the type strain of which is CCUG 48324(T) (=CIP 108195(T)).

Streptococcus marimammalium sp nov., isolated from seals

Two strains of an unidentified, Gram-positive, catalase-negative, chain-forming, coccus-shaped organism recovered from seals were characterized using phenotypic and molecular taxonomic methods. Based on morphological and biochemical criteria the strains were tentatively identified as streptococci but they did not appear to correspond to any recognized species of the genus Streptococcus. Comparative 16S rRNA gene sequencing studies showed that the strains were closely related to each other and confirmed their placement in the genus Streptococcus. Sequence divergence values of > 5 % with reference streptococcal species demonstrated the organisms from seals represent a novel species. SDS-PAGE analysis of whole-cell proteins confirmed that the two organisms were closely related to each other but were different from all currently defined streptococcal species. Based on biochemical criteria, molecular chemical and molecular genetic evidence, it is proposed that the unknown isolates from seals be assigned to a novel species of the genus Streptococcus, Streptococcus marimammalium sp. nov. The type strain is M54/01/(T) (=CCUG 48494(T)=CIP 108309(T)).

Streptococcus equi subsp ruminatorum subsp nov., isolated from mastitis in small ruminants

Six isolates of an unknown Gram-positive, catalase-negative, chain-forming, coccus-shaped organism isolated from ovine and caprine mastitis were characterized by phenotypic and molecular taxonomic methods. On the basis of cellular morphology and the results of biochemical tests, the organism was tentatively identified as a streptococcal species. Comparative 16S rRNA gene sequencing studies confirmed that the organism is a member of the genus Streptococcus, with Streptococcus equi as its closest phylogenetic relative (98(.)8% similarity). DNA-DNA pairing studies showed that the unidentified organism displayed more than 70% relatedness to the type strains of S. equi subsp. equi and subsp. zooepidemicus. Despite the relatively high DNA-DNA reassociation values, biotyping and ribotyping allowed clear differentiation of the unknown bacterium from the two recognized subspecies of S. equi. On the basis of phenotypic and molecular genetic evidence, it is proposed that the unknown Streptococcus isolates from ovine and caprine mastitis be classified as a novel subspecies, Streptococcus equi subsp. ruminatorum subsp. nov. The type strain is CECT 5772(T) (=CCUG 47520(T) = Mt 167(T)).

Streptococcus devriesei sp nov., from equine teeth

Phenotypic and phylogenetic studies were performed on four unidentified Gram-positive staining, catalase-negative, cc-hemolytic Streptococcus-like organisms recovered from the teeth of horses. SDS PAGE analysis of whole-cell proteins and comparative 16S rRNA gene sequencing demonstrated the four strains were highly related to each other but that they did not correspond to any recognised species of the genus Streptococcus. Phylogenetic analysis based on 16S rRNA gene sequences showed the unidentified organisms form a hitherto unknown sub-line within the Streptococcus genus, displaying a close affinity with Streptococcus mutans, Streptococcus ferus and related organisms. Sequence divergence values of > 5% with thew and other reference streptococcal species however demonstrated the organisms from equine sources represent a novel species. Based on the phenotypic distinctiveness of the new bacterium and molecular chemical and molecular genetic evidence, it is proposed that the unknown species be classified as Streptococcus devriesei sp. nov. The type strain of Streptococcus devriesei is CCUG 47155(T) (= CIP 107809T).

Streptococcus ovis sp. nov., isolated from sheep

Seven strains of an unknown Gram-positive catalase-negative chain-forming coccus-shaped organism isolated from clinical specimens from sheep were characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing studies demonstrated that the bacterium represents a new sub-line within the genus Streptococcus. The unknown bacterium was readily distinguished from recognized streptococcal species by biochemical tests and electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, it is proposed that the unknown bacterium be classified as Streptococcus ovis sp. nov. The type strain of Streptococcus ovis is CCUG 39485T (= LMG 19174T).

In vitro immunomodulatory activity of Lactobacillus fermentum CECT5716 and Lactobacillus salivarius CECT5713: two probiotic strains isolated from human breast milk

Commensal bacteria, including some species of lactobacilli commonly present in human breast milk, appear to colonize the neonatal gut and contribute to protection against infant infections, suggesting that lactobacilli could potentially modulate immunity. In this study, we evaluated the potential of two Lactobacillus strains isolated from human milk to modulate the activation and cytokine profile of peripheral blood mononuclear cell (PBMC) subsets in vitro. Moreover, these effects were compared to the same probiotic species of non-milk origin. Lactobacillus salivarius CECT5713 and Lactobacillus fermentum CECT5716 at 105, 106 and 107 bacteria/mL were co-cultured with PBMC (106/mL) from 8 healthy donors for 24 h. Activation status (CD69 and CD25 expressions) of natural killer (NK) cells (CD56+), total T cells (CD3+), cytotoxic T cells (CD8+) and CD4+ T cells was determined by flow cytometry. Regulatory T cells (Treg) were also quantified by intracellular Foxp3 evaluation. Regarding innate immunity, NK cells were activated by addition of both Lactobacillus strains, and in particular, the CD8+ NK subset was preferentially induced to highly express CD69 (90%, p<0.05). With respect to acquired immunity, approximately 9% of CD8+ T cells became activated after co-cultivation with L. fermentum or L salivarius. Although CD4+ T cells demonstrated a weaker response, there was a preferential activation of Treg cells (CD4+CD25+Foxp3+) after exposure to both milk probiotic bacteria (p<0.05). Both strains significantly induced the production of a number of cytokines and chemokines, including TNFα, IL-1β, IL-8, MIP-1α, MIP-1β, and GM-CSF, but some strain-specific effects were apparent. This work demonstrates that L salivarius CECT5713 and L. fermentum CECT5716 enhanced both natural and acquired immune responses, as evidenced by the activation of NK and T cell subsets and the expansion of Treg cells, as well as the induction of a broad array of cytokines.

Induction of neutralizing antibodies in mice immunized with an amino-terminal polypeptide of Streptococcus mutans P1 protein produced by a recombinant Bacillus subtilis strain

The oral pathogen Streptococcus mutans expresses a surface protein, P1, which interacts with the salivary pellicle on the tooth surface or with fluid-phase saliva, resulting in bacterial adhesion or aggregation, respectively. P1 is a target of protective immunity. Its N-terminal region has been associated with adhesion and aggregation functions and contains epitopes recognized by efficacious antibodies. In this study, we used Bacillus subtilis, a gram-positive expression host, to produce a recombinant N-terminal polypeptide of P1 (P1(39-512)) derived from the S. mutans strain UA159. Purified P1(39-512) reacted with an anti-full-length P1 antiserum as well as one raised against intact S. mutans cells, indicating preserved antigenicity. Immunization of mice with soluble and heat-denatured P1(39-512) induced antibodies that reacted specifically with native P1 on the surface of S. mutans cells. The anti-P1(39-512) antiserum was as effective at blocking saliva-mediated aggregation of S. mutans cells and better at blocking bacterial adhesion to saliva-coated plastic surfaces compared with the anti-full-length P1 antiserum. In addition, adsorption of the anti-P1 antiserum with P1(39-512) eliminated its ability to block the adhesion of S. mutans cells to abiotic surfaces. The present results indicate that P1(39-512), expressed and purified from a recombinant B. subtilis strain, maintains important immunological features of the native protein and represents an additional tool for the development of anticaries vaccines.

Nasal immunization of mice with Lactobacillus casei expressing the Pneumococcal Surface Protein A: induction of antibodies, complement deposition and partial protection against Streptococcus pneumoniae challenge

Strategies for the development of new vaccines against Streptococcus pneumoniae infections try to overcome problems such as serotype coverage and high costs, present in currently available vaccines. Formulations based on protein candidates that can induce protection in animal models have been pointed as good alternatives. Among them, the Pneumococcal Surface Protein A (PspA) plays an important role during systemic infection at least in part through the inhibition of complement deposition on the pneumococcal surface, a mechanism of evasion from the immune system. Antigen delivery systems based on live recombinant lactic acid bacteria (LAB) represents a promising strategy for mucosal vaccination, since they are generally regarded as safe bacteria able to elicit both systemic and mucosal immune responses. In this work, the N-terminal region of clade I PspA was constitutively expressed in Lactobacillus casei and the recombinant bacteria was tested as a mucosal vaccine in mice. Nasal immunization with L. casei-PspA 1 induced anti-PspA antibodies that were able to bind to pneumococcal strains carrying both clade 1 and clade 2 PspAs and to induce complement deposition on the surface of the bacteria. In addition, an increase in survival of immunized mice after a systemic challenge with a virulent pneumococcal strain was observed. (C) 2008 Elsevier Masson SAS. All rights reserved.

Estudo da resistência do streptococcus pneumoniae à penicilina em pneumopatias infecciosas nas cidades de Porto Alegre e Caxias do Sul (RS - Brasil)

A resistência aos antibióticos dos patógenos mais comuns do trato respiratório está aumentando mundialmente. Recentemente, Streptococcus pneumoniae resistente à penicilina tem sido isolado em diversos países, e a freqüência dessas cepas tem elevado de modo alarmante. O aumento da resistência, com conseqüentes implicações terapêuticas, tem levado a uma reavaliação do uso dos antibióticos ß-lactâmicos para o tratamento de infecções pneumocócicas. No presente trabalho, um total de 107 amostras de Streptococcus pneumoniae, obtidas de materiais provenientes de pacientes adultos ambulatoriais e hospitalizados, em dois centros médicos de duas cidades do Rio Grande do Sul (Porto Alegre e Caxias do Sul), os quais apresentavam quadro clínico-radiológico de infecção pulmonar, foram analisadas com o objetivo de estudar-se a resistência do germe à penicilina. As amostras constituídas de escarro (80,4%), lavado brônquico (13,5%) e aspirado traqueal (6,6%) foram coletadas no período compreendido entre Julho de 1998 e Julho de 1999. O material foi semeado em meio de Agar sangue e as colônias suspeitas de Streptococcus pneumoniae foram transferidas para meio de Mueller-Hinton para teste de optoquina e de sensibilidade à penicilina com discos de oxacilina. Um halo de inibição da oxacilina menor do que 20 mm indicava a realização de teste para determinação da concentração inibitória mínima (MIC) com E-test. Um total de nove cepas foi identificado como tendo resistência intermediária à penicilina (MIC 0,1-1,0μg/ml) e nenhuma cepa resistente (resistência elevada: MIC > 2,0 μg/ml) foi identificada. Uma monitorização local das cepas quanto à resistência antimicrobiana é de grande importância para os clínicos no manejo de infecções pneumocócicas.

Isolamento e resistência a antimicrobianos de cepas de Streptococcus spp. provenientes de rãs-touro (Lithobates catesbeianus)

Twelve bullfrogs were selected from two commercial frog farms and clinically diagnosed as attacked by Streptococcus disease. Sixty samples were collected, and Streptococcus spp. was isolated from all bullfrog, being 12 (100%) from the encephalus, seven from the kidneys (58.3%), three from the liver (25%), two from the spleen (16.6%), and one from the ascitic liquid (8.3%). Streptococcus -hemolytic were isolated from all the 60 samples, which were sensible to chloramphenicol (100%), gentamycin (100%), vancomycin (96%), cefotaxime (96%), and cefoxitine (92%).

Inflammatory responses of Nile tilapia Oreochromis niloticus to Streptococcus agalactiae: effects of vaccination and yeast diet supplement

This study evaluated the effects of dietary supplementation with 0.3% Saccharomyces cerevisiae yeast cell wall and of vaccination against Streptococcus agalactiae on the cellular component of acute inflammation induced in the coelomic cavity of Nile tilapia Oreochromis niloticus and on survival of the fish after challenge. A total of 84 tilapia of mean (+/- SD) weight 125.0 +/- 1.5 g were distributed among twelve 310 l fiberglass tanks according to a 2 x 2 x 3 factorial design in the following manner: with and without supplementation; 2 stimulations (oily solution without S. agalactiae vaccine and vaccination); 15 d later all fish were intracoelomically challenged with 10(8) CFU ml(-1) of a homologous strain of S. agalactiae, and evaluated after 6, 24 and 48 h, with 7 replicates. The fish received the non-supplemented or supplemented diet for a total of 77 d. The vaccination was performed on the 60th day, intracoelomically, as a single injection of 0.5 ml of the vaccine containing 10(8) CFU ml(-1). Fifteen days later, all the fish were challenged with S. agalactiae by means of an intracoelomic inoculation of 10(8) CFU ml(-1). No mortality was observed among the supplemented fish. The fish that were fed the non-supplemented diet and immunized with the bacterium presented a mortality rate of 28.5%. Among the non-supplemented and non-immunized fish, the mortality rate was 38.09%. Supplementation, in both vaccinated and non-vaccinated fish, induced larger accumulations of thrombocytes, lymphocytes and macrophages at the inflammatory focus. The results suggest that supplementation with 0.3% yeast cell wall, in both vaccinated and non-vaccinated fish, improved the inflammatory response of the fish and protected against the challenge. Vaccination increased the defense response, but the effect was stronger when associated with supplementation with S. cerevisiae.