Three unusual cases of parasites in eye.

A 17-year-old male patient presented with cellulitis and mass in the eye noticed approximately 3 months back. The mass was about 1 cm in size and situated at the limbus. All preoperative routine investigations were normal. Surgical exploration revealed a sub-conjunctival cystic mass near the lateral rectus muscle about 1 cm in diameter; the mass was excised. Gross pathological examination revealed a thin-walled cystic mass. There was a hard nodule in the center. Microscopy revealed a wall of cysticercosis. Scolex was also seen. Surrounding tissue revealed sparse acute and chronic inflammatory cells. The case was confirmed by CDC, Atlanta, and was also included in their departmental presentation as an interesting case. A 60-year-old lady presented with complaints of itching over the forehead and right eye for 5 days. She was prescribed steroid eyedrops and antihistaminics. The itching aggravated with eyedrops along with watering and foreign body sensation. On revisit, the ophthalmologist noticed a worm in the right upper subconjunctival space. The worm was carefully removed in toto and sent to the laboratory for identification. The worm was thin, cylindrical, 8-10 cm long and white in color. After microscopic and gross examination of the worm, it was identified as Dirofilaria spp. CDC (Atlanta) confirmed the diagnosis of Dirofilaria. The patient was treated with antihistaminics and was relieved of symptoms without recurrence. A 45-year-old male patient had a painless mass in the eye for the last 3 months. He had no systemic illness. He gave a history of swimming pool use during that time. The mass was excised and submitted for histopathology. Numerous globular cysts representing thick-walled sporangia containing numerous spores diagnostic of Rhinosporidiosis were seen.

Elusive treatment for human rhinosporidiosis.

OBJECTIVES: The aim of this study was to clarify the contentious taxonomic classification of Rhinosporidium seeberi, the cause of human rhinosporidiosis, which may have treatment implications. METHODS: PCR was used to amplify the internal transcribed spacer (ITS)-2 region from the genomic DNA of the aetiological agent obtained from a sample of human rhinosporidiosis lesions. The amplicon was sequenced and the organism identified using the Basic Local Alignment Search Tools (BLAST). RESULTS: Phylogenetic analysis revealed that the aetiological agent clustered along with the R. seeberi isolated from humans and also with Amphibiocystidium ranae from frogs. This organism is a member of the order Dermocystida in the class Mesomycetozoea. A patient with disseminated rhinosporidiosis did not respond to conventional therapy with dapsone and surgical excision, and treatment with amphotericin B also proved futile. CONCLUSION: An effective treatment for R. seeberi-a eukaryote belonging to the class Mesomycetozoea-is still elusive.

Rhinosporidiosis in southwest Bengal.

Rhinosporidiosis is a non-contagious chronic granulomatous disease that is prevalent in southern India and Sri Lanka. It has been known for centuries, but the details of the disease and the precise manner of its transmission have, until recently, remained unknown. Our institution sees many cases of this disease and we investigate the management protocol and its recent advances and include a review of the published literature. A total of 152 patients who were treated at Bankura Sammilani Medical College were studied between 2005 and 2011. The most common age group affected were those aged between 11 and 20 years of age and the male-to-female ratio was 1.9:1. Three patients suffered recurrent disease - one experienced it on the same site and the others on distant sites. Eleven patients with inadequate excision in which the margins were not free from disease were treated with dapsone therapy without any reported recurrence. It is a common disease in southwestern West Bengal. Surgical excision with electrocoagulation of the base is the main treatment, and dapsone therapy is recommended in order to prevent recurrences in multiple sites of affection and inadequate surgically excised cases. Although the disease occurs sporadically in most parts of the world, we see many patients in our area.

Recovery of growth of Hyphochytrium catenoides after exposure to environmental stress.

The survival of an isolate of Hyphochytrium catenoides collected from soil in the Blue Mountains in eastern New South Wales, Australia, was tested under extreme conditions in the laboratory. This isolate recovered growth after being subjected to drying on filter paper, to heat while desiccated, to hypersalinity, to strict anaerobic conditions, to freezing temperatures, and to a short period in solutions at pH 2.8-11.2. The capacity to survive under these conditions in the laboratory suggests adaptation to fluctuating conditions in the soil. The partial DNA sequence of the 28S ribosomal RNA gene in the isolate from New South Wales was 98% similar to that in an isolate from Arizona with a similar morphology.

The effects of biocides (antiseptics and disinfectants) on the endospores of Rhinosporidium seeberi.

No data exists on the activity of biocides (antiseptics and disinfectants) on Rhinosporidium seeberi that causes rhinosporidiosis in humans and animals. On account of the inability to culture R. seeberi, in vitro, dyes were used to assess the morphological integrity and viability of biocide-treated endospores that are considered to be the infective stage of this pathogen. Evan's Blue (EvB) identifies the morphological integrity of the endospores while MTT (3-[4, 5-dimethylthiazol-2yl]-2, 5-diphenyl tetrazolium bromide) identifies metabolic activity through its reduction by cellular dehydrogenases to microscopically visible deposits of insoluble formazan. MTT-negativity has earlier been shown to correlate with absence of growth of yeast and mycelial fungi in culture and could thus indicate the loss of viability of MTT-negative rhinosporidial endospores. Hydrogen peroxide, glutaraldehyde, chloroxylenol, chlorhexidine, cetrimide, thimerosal, 70% ethanol, iodine in 70% ethanol, 10% formalin, povidone-iodine, sodium azide and silver nitrate were tested on freshly-harvested endospores and all biocides caused metabolic inactivation with or without altered structural integrity as shown by absence of MTT-staining after 3, 24 or 36 hour after exposure, while EvB stained only the endospores treated with sodium azide, ethanol, thimerosal, chloroxylenol, glutaraldehyde and hydrogen peroxide. With clinically useful biocides - chlorhexidine, cetrimide-chlorhexidine, 70% ethanol, povidone-iodine and silver nitrate, a total period of exposure of endospores to the biocide, for seven minutes, produced metabolic inactivation of the endospores. Anti-rhinosporidial antiseptics that could be used in surgery on rhinosporidial patients include povidone-iodine in nasal packs for nasal and naso-pharyngeal surgery, chlorhexidine and cetrimide-chlorhexidine on the skin, while povidone-iodine and silver nitrate could have application in ocular rhinosporidiosis.

Human anti-rhinosporidial antibody does not cause metabolic inactivation or morphological damage in endospores of Rhinosporidium seeberi, in vitro.

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.

Light and electron microscopic findings in rhinosporidiosis after dapsone therapy.

Morphological findings in serial nasal mucosal biopsies from three cases of rhinosporidiosis on dapsone therapy were compared with biopsies from 33 patients taken before dapsone or surgical treatment was initiated. All biopsies were examined by light microscopy and five by electron microscopy. Counts of histologically intact and degenerated organisms showed a decreasing proportion of intact forms with treatment.