Bats are a unique group of mammals well suited to be hosts for emerging viruses. With current rates of deforestation and urbanization, redistribution of bat habitats to urban and suburban areas may bring bats into closer contact with livestock and humans. Common flying fox, Pteropus medius (previously known as Pteropus giganteus), forms large communal roosts on treetops, often in close proximity to human habitation in Sri Lanka. This report describes the detection of coronavirus RNA in P. medius bat guano collected in Peradeniya, Sri Lanka. These viruses had >97% nucleotide identity with coronaviruses detected in Cynopterus sphinx, Scotophilus heathii and S. kuhlii bats in Thailand. Pteropus medius is widespread in Asia and appears to excrete group D coronaviruses, which are hitherto confined to bats; however, these findings may have public health implications in the future.
This report describes the use of the MTT-reduction and Evan's blue-staining tests for the assessment of the viability and morphological integrity, respectively, of rhinosporidial endospores after exposure to sera from rhinosporidial patients with high titres of anti-rhinosporidial antibody. Sera from three patients, with nasal, ocular and disseminated rhinosporidiosis respectively were used, with human serum without anti-rhinosporidial antibody for comparison, with or without added fresh guinea pig serum as a source of complement. All four sera tested, with or without guinea-pig serum, had no effect on the morphological integrity or the viability of the endospores and it is suggested that anti-rhinosporidial antibody has no direct protective role against the endospores, the infective stage, in rhinosporidiosis. This finding is compatible with the occurrence of chronicity, recurrence and dissemination that are characteristic of rhinosporidiosis despite the presence of high titres of anti-rhinosporidial antibody in patients with these clinical characteristics. The possible occurrence of humoral mechanisms of immunity that involve anti-rhinosporidial antibody with cells such as leucocytes and NK cells, in vivo, cannot yet be discounted, although the presence of high titres of anti-rhinosporidial antibody in patients with chronic, recurrent and disseminated lesions might indicate that such antibody is non-protective in vivo.
Introduction Non-tuberculous mycobacteria (NTM) are known to cause opportunistic nosocomial infections. The aim of the study was to identify NTM using culture characteristics, biochemical and molecular methods (multiplex PCR) from stored mycobacteria isolated from patients presenting with pulmonary and extrapulmonary tuberculosis at Teaching Hospitals, Kandy and Peradeniya during a 4-year period from 2004 to 2009. Methods Forty mycobacterial isolates obtained from 48 samples of sputum, urine, broncho-alveolar lavage and peritoneal fluid were analyzed initially by growth characteristics, followed by biochemical and molecular methods for the presence of NTM. Five main growth characteristics were analyzed, which included rate of growth, growth-temperature relationship, production of pigment, colony morphology and growth on McConkey agar. Species identification of these NTM isolates was by performing biochemical tests. Molecular identification was performed by multiplex-PCR technique on mycobacterial clinical isolates for the rapid identification of NTM. Results Of the 40 mycobacterial isolates, 10 were identified as NTM by using culture characteristics and multiplex PCR. All the NTM isolates were identified to species level by biochemical methods and eight species of NTM were identified. These species, obtained from sterile as well as non-sterile clinical samples were identified as M. scrofulaceum, M. haemophilum, M. chelonae, M. kansasii, M. phlei, M. flavescens, M. gastri and M. vaccae. Conclusion Non-tuberculous mycobacteria were isolated from a wide range of sterile and non-sterile clinical samples and contributed to 25% of the mycobacterial isolates in this study. The results of culture characteristics were compatible with the molecular identification (multiplex PCR) differentiating Mycobacterium tuberculosis from non-tuberculous mycobacterial strains. M. kansasii and M. gastri were the most common NTM isolated from clinical isolates.
Human rhinosporidial tissue was used as the source of the various developmental stages of Rhinosporidium seeberi--endospores with electron dense bodies, juvenile, and immature sporangia. After homogenisation in phosphate buffered saline (PBS) and removal of tissue fragments by centrifugation, the rhinosporidial bodies were isolated on centrifuged Percoll columns with gradients of densities or on triple-layered columns of varying density. The separated bands, after repeated washing in PBS gave bodies free from human tissue as shown on Leishman and PAS staining and indirect immunofluorescence with rabbit and human patients' anti-rhinosporidial sera. Sonicates of these bodies were tested on agarose gel for precipitation with antisera, and on SDS-PAG electrophoresis and Coomassie Blue staining. Percoll columns were shown to be capable of isolating these stages of R. seeberi, free from human tissue and contaminating bacteria.
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