Characterization of bacteriophages infecting clinical isolates of Pseudomonas aeruginosa stored in a culture collection.

Some clinical isolates of Pseudomonas aeruginosa stored in our culture collection did not grow or grew poorly and showed lysis on the culture plates when removed from the collection and inoculated on MacConkey agar. One hypothesis was that bacteriophages had infected and killed those clinical isolates. To check the best storage conditions to maintain viable P. aeruginosa for a longer time, clinical isolates were stored at various temperatures and were grown monthly. We investigated the presence of phage in 10 clinical isolates of P. aeruginosa stored in our culture collection. Four strains of P. aeruginosa were infected by phages that were characterized by electron microscopy and isolated to assess their ability to infect. The best condition to maintain the viability of the strains during storage was in water at room temperature. Three Siphoviridae and two Myoviridae phages were visualized and characterized by morphology. We confirmed the presence of bacteriophages infecting clinical isolates, and their ability to infect and lyse alternative hosts. Strain PAO1, however, did not show lysis to any phage. Mucoid and multidrug resistant strains of P. aeruginosa showed lysis to 50% of the phages tested.

Characterization of bacteriophages infecting clinical isolates of Pseudomonas aeruginosa stored in a culture collection

Some clinical isolates of Pseudomonas aeruginosa stored in our culture collection did not grow or grew poorly and showed lysis on the culture plates when removed from the collection and inoculated on MacConkey agar. One hypothesis was that bacteriophages had infected and killed those clinical isolates. To check the best storage conditions to maintain viable P. aeruginosa for a longer time, clinical isolates were stored at various temperatures and were grown monthly. We investigated the presence of phage in 10 clinical isolates of P. aeruginosa stored in our culture collection. Four strains of P. aeruginosa were infected by phages that were characterized by electron microscopy and isolated to assess their ability to infect. The best condition to maintain the viability of the strains during storage was in water at room temperature. Three Siphoviridae and two Myoviridae phages were visualized and characterized by morphology. We confirmed the presence of bacteriophages infecting clinical isolates, and their ability to infect and lyse alternative hosts. Strain PAO1, however, did not show lysis to any phage. Mucoid and multidrug resistant strains of P. aeruginosa showed lysis to 50% of the phages tested.

Influenza virus A(H3N2) strain isolated from cerebrospinal fluid from a patient presenting myelopathy post infectious.

BACKGROUND: Neurological involvement during influenza infection has been described during epidemics and is often consistent with serious sequelae or death. OBJECTIVE: To investigate the etiologic agent involved in myelopathy post influenza-like syndrome. STUDY DESIGN: This investigation focuses on virus isolation from the cerebrospinal fluid (CSF) collected from a 19-year-old male student presenting with clinical diagnosis of myelopathy post influenza-like syndrome. To achieve this goal, different cell cultures and molecular methodologies were carried out. RESULTS: Influenza virus A(H3N2) strain was isolated in MDCK cell culture; virus particles were observed under electron microscopy. Phylogenetics analyses showed that the Brazilian influenza A(H3N2) strains were closely related to the A/Perth/16/2009-like. CONCLUSION: This study demonstrates that influenza virus A(H3N2) strain was the cause of illness of the students. According to the Brazilian influenza virus sentinel surveillance data A/Perth/16/2009-LIKE (H3N2) strain has predominated during the 2010 influenza virus season in Brasília-DF.

Diversity of somatic coliphages in coastal regions with different levels of anthropogenic activity in São Paulo State, Brazil.

Bacteriophages are the most abundant and genetically diverse viruses on Earth, with complex ecology in both quantitative and qualitative terms. Somatic coliphages (SC) have been reported to be good indicators of fecal pollution in seawater. This study focused on determining the concentration of SC and their diversity by electron microscopy of seawater, plankton, and bivalve samples collected at three coastal regions in São Paulo, Brazil. The SC counts varied from <1 to 3.4 × 10(3) PFU/100 ml in seawater (73 samples tested), from <1 to 4.7 × 10(2) PFU/g in plankton (46 samples tested), and from <1 to 2.2 × 10(1) PFU/g in bivalves (11 samples tested). In seawater samples, a relationship between the thermotolerant coliforms and Escherichia coli and SC was observed at the three regions (P = 0.0001) according to the anthropogenic activities present at each region. However, SC were found in plankton samples from three regions: Baixada Santista (17/20), Canal de São Sebastião (6/14), and Ubatuba (3/12). In seawater samples collected from Baixada Santista, four morphotypes were observed: A1 (4.5%), B1 (50%), C1 (36.4%), and D1 (9.1%). One coliphage, Siphoviridae type T1, had the longest tail: between 939 and 995 nm. In plankton samples, Siphoviridae (65.8%), Podoviridae (15.8%), Microviridae (15.8%), and Myoviridae (2.6%) were found. In bivalves, only the morphotype B1 was observed. These SC were associated with enteric hosts: enterobacteria, E. coli, Proteus, Salmonella, and Yersinia. Baixada Santista is an area containing a high level of fecal pollution compared to those in the Canal de São Sebastião and Ubatuba. This is the first report of coliphage diversity in seawater, plankton, and bivalve samples collected from São Paulo coastal regions. A better characterization of SC diversity in coastal environments will help with the management and evaluation of the microbiological risks for recreation, seafood cultivation, and consumption.

An outbreak of group C rotavirus gastroenteritis among adults living in Valentim Gentil, São Paulo State, Brazil.

An outbreak of gastroenteritis affecting adults and children occurred in the small city of Valentim Gentil, São Paulo, Brazil, in 1993. Nineteen faecal samples (from 10 cases and 9 contracts) were examined by direct electron microscopy (DEM), immune electron microscopy (IEM), polyacrylamide gel electrophoresis (PAGE), and enzyme-linked immunosorbent assay (ELISA) for group A and C rotaviruses. DEM detected rotavirus in 6 of the 10 cases and in none of the contacts. All of the samples were negative for group A rotavirus by ELISA. Analysis by PAGE showed an electrophoretic profile suggestive of group C rotavirus in two cases. Group C rotavirus was identified by IEM in 4 of the cases and in 1 of the contacts. All of the samples were submitted to ELISA for group C rotavirus. This resulted in a total of 10 positives-7 for diarrhoeal cases and 3 for contacts. This outbreak was strongly associated with group C rotavirus. The importance of combining different diagnostic methods is emphasised.

Intrafamilial outbreak of astrovirus gastroenteritis in São Paulo, Brazil.

This report describes an intrafamilial outbreak of astrovirus-associated gastroenteritis that occurred among 5 children in a family living in the State of São Paul, Brazil. Astrovirus was detected by direct electron microscopy (DEM) and confirmed by solid phase immune electron microscopy (SPIEM) using astrovirus reference serum (T2-6-89). The rise in titre of astrovirus antibody in patients' sera was observed by IEM using acute and convalescent-phase sera. Combined use of the EM techniques to detect virus in stool and antibody rise in sera led us to conclude that this intrafamilial outbreak might have been caused by astrovirus infection. In Brazil, astrovirus may be a more common agent of gastroenteritis than in currently appreciated.

[Rotavirus, adenovirus, astrovirus, calicivirus and small round virus particles in feces of children with and without acute diarrhea, from 1987 to 1988, in the greater São Paulo]. / Rotavírus, adenovírus, astrovírus, calicivírus e "small round virus particles" em fezes de crianças, com e sem diarréia aguda, no período de 1987 a 1988, na grande São Paulo.

Between 1987 and 1988, 193 faecal specimens from children, with or without diarrhea, were submitted to enzyme immunoassay, polyacrylamide-gel electrophoresis and electronmicroscopy tests for virus detection. The positivity for Rotavirus, Adenovirus, Astrovirus, Calicivirus and Small Round Virus Particles (SRVP) was 11.3%, 3.1%, 2.1%, 1.0% and 4.1%, respectively, for the 97 children with acute diarrhea. Of the 96 children without diarrhea, 4.2% were positive for Rotavirus, 1.0% for Calicivirus and 7.3% for SRVP. Of 15 positive specimens for Rotavirus, 14 showed electrophoretic patterns proper to group A and 1 specimen of group C Rotavirus. The analysis of electrophorotypes demonstrated great heterogeneity of electrophoretic patterns and predominance of subgroup 2, "long". The association of virus, bacteria and parasites was present both in children with or without acute diarrhea.

Occurrence of retrovirus-like particles in various cellular and intercellular compartments of the venom glands from Bothrops jararacussu.

Retrovirus-like particles were detected in venom glands from Bothrops jararacussu during electron microscopy. Type C-like particles were found inter- and intracellularly in gland and vessel lumina and scattered in the connective tissue. They were about 100 nm in diameter, with an electron dense core and bilaminar external membrane. Shapes suggestive of a budding process from the plasma membrane were also observed. Less frequently, type A-like particles, about 80 nm diameter with an electronlucent core, appeared in association with the membranes of the endoplasmic reticulum of the secretory cells.