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D V Seal

Publications and source records attributed to D V Seal.

At least 19 recordsLinked to original sources
PubMed2002

18S ribosomal DNA typing and tracking of Acanthamoeba species isolates from corneal scrape specimens, contact lenses, lens cases, and home water supplies of Acanthamoeba keratitis patients in Hong Kong.

We examined partial 18S ribosomal DNA (Rns) sequences of Acanthamoeba isolates cultured in a study of microbial keratitis in Hong Kong. Sequence differences were sufficient to distinguish closely related strains and were used to examine links between strains obtained from corneal scrape specimens, contact lenses, lens cases, lens case solutions, and home water-supply faucets of patients with Acanthamoeba. We also looked for evidence of mixed infections. Identification of Acanthamoeba Rns genotypes was based on sequences of approximately 113 bp within the genus-specific amplicon ASA.S1. This permitted genotype identification by using nonaxenic cultures. Of 13 specimens obtained from corneal scrapes, contact lenses, lens cases, or lens case solutions, 12 were Rns genotype T4 and the remaining one was Rns genotype T3. The sequences of corneal scrape specimens of two patients also were the same as those obtained from their contact lenses or lens case specimens. A possible triple-strain infection was indicated by three different T4 sequences in cultures from one patient's lenses. Although faucet water used by patients to clean their lenses is a possible source of infections, specimens isolated from the faucets at two Acanthamoeba keratitis patients' homes differed from their corneal scrape or lens specimens. The overall results demonstrate the potential of this Rns region for tracking Acanthamoeba keratitis strains in infections and for distinguishing single-strain and closely related multiple-strain infections even when other microorganisms might be present with the cultured specimens. They also confirm the predominance of Rns genotype T4 strains in Acanthamoeba keratitis infections.

Acanthamoeba↗
PubMed2001

Necrotizing fasciitis.

Necrotizing fasciitis continues to occur due to beta-haemolytic streptococci but is now also recognized as being due to Vibrio spp. in fishermen and those in contact with warm water in the Gulf of Mexico and South-East Asia, including Hong Kong. Magnetic resonance image scanning has identified the extent of fasciitis and soft tissue oedema infiltrating fascial planes prior to necrosis presenting clinically and is a useful tool in early diagnosis. Surgical debridement or incisional drainage remains essential. An enhanced bactericidal response against beta-haemolytic streptococci has been found with a combination of penicillin and clindamycin. Intravenous immunoglobulin has been shown to reduce mortality if the necrotizing fasciitis is associated with the toxic shock syndrome, by decreasing the superantigen activity of the beta-haemolytic streptococci on cytokine release by T cells.

Adult↗
PubMed2001

Use of subgenic 18S ribosomal DNA PCR and sequencing for genus and genotype identification of acanthamoebae from humans with keratitis and from sewage sludge.

This study identified subgenic PCR amplimers from 18S rDNA that were (i) highly specific for the genus Acanthamoeba, (ii) obtainable from all known genotypes, and (iii) useful for identification of individual genotypes. A 423- to 551-bp Acanthamoeba-specific amplimer ASA.S1 obtained with primers JDP1 and JDP2 was the most reliable for purposes i and ii. A variable region within this amplimer also identified genotype clusters, but purpose iii was best achieved with sequencing of the genotype-specific amplimer GTSA.B1. Because this amplimer could be obtained from any eukaryote, axenic Acanthamoeba cultures were required for its study. GTSA.B1, produced with primers CRN5 and 1137, extended between reference bp 1 and 1475. Genotypic identification relied on three segments: bp 178 to 355, 705 to 926, and 1175 to 1379. ASA.S1 was obtained from single amoeba, from cultures of all known 18S rDNA genotypes, and from corneal scrapings of Scottish patients with suspected Acanthamoeba keratitis (AK). The AK PCR findings were consistent with culture results for 11 of 15 culture-positive specimens and detected Acanthamoeba in one of nine culture-negative specimens. ASA.S1 sequences were examined for 6 of the 11 culture-positive isolates and were most closely associated with genotypic cluster T3-T4-T11. A similar distance analysis using GTSA.B1 sequences identified nine South African AK-associated isolates as genotype T4 and three isolates from sewage sludge as genotype T5. Our results demonstrate the usefulness of 18S ribosomal DNA PCR amplimers ASA.S1 and GTSA.B1 for Acanthamoeba-specific detection and reliable genotyping, respectively, and provide further evidence that T4 is the predominant genotype in AK.

Acanthamoeba↗
PubMed2000

Salicylate inhibition of Acanthamoeba attachment to contact lenses: a model to reduce risk of infection.

PURPOSE: Acanthamoeba attachment (adsorption) to hydrogel contact lenses is enhanced by Pseudomonas aeruginosa biofilm. The effect of sodium salicylate on Acanthamoeba attachment to biofilm-coated and uncoated hydrogel lenses was investigated. DESIGN: Experimental study. PARTICIPANTS AND CONTROLS: A minimum of 16 replicates were used for each test condition; a control condition using clean lenses without biofilm was included. METHODS: Four groups of hydrogel contact lenses (etafilcon A) were pretreated with P. aeruginosa to form a biofilm. In addition, two more groups remained untreated. Quartered lenses of all six groups were then incubated in a suspension of A. castellanii trophozoites. Two batches of lenses had either 3 or 30 mM sodium salicylate added to the bacterial suspension (stage 1 intervention). Two other batches of lenses had salicylate added to the amoebal suspension (stage 2 intervention). One of the batches, which had a stage 1 intervention, had salicylate added at the second stage as well. The remaining batches received no salicylate exposure and included lenses with and without biofilm coating. MAIN OUTCOME MEASURE: The outcome measure in this study was the number of Acanthamoeba trophozoites attached, per square centimeter, to the hydrogel surfaces. RESULTS: Biofilm coating from P. aeruginosa gave a significantly increased attachment of A. castellanii trophozoites to the contact lens. When introduced at a first (biofilm) stage, second (trophozoite attachment) stage, or with intervention at both stages, 30 mM sodium salicylate reduced amoebal attachment to the hydrogel lens. When applied to both stages and when applied at stage 2 to the biofilm coated contact lenses, 3 mM sodium salicylate reduced amoebal attachment. The 3 mM concentration was not effective for the lower level of amoebae attachment to uncoated (nonbiofilm) lenses. CONCLUSIONS: Sodium salicylate successfully reduced amoebal trophozoite attachment to hydrogel lenses. This was the result of one of the following possibilities or a combination thereof: inhibition of biofilm formation; a direct effect on the amoebae; an alteration in the biofilm-amoebal attachment and resulting modification of the hydrogel lens surface. The results of this study suggest the major action is at stage 2 (on amoebal attachment to lenses) and favors alteration of the biofilm-amoebal attachment mechanism. This study demonstrates salicylate's potential benefit as a component of contact lens care solutions, designed to reduce microbial attachment and the risk of infection.

Acanthamoeba↗
PubMed1999

Disinfection of contact lenses without tap water rinsing: is it effective?

PURPOSE: To establish the efficacy of the two most popular contact lens disinfecting systems--one-step hydrogen peroxide and multipurpose disinfecting solution--for 1 month's use in practice in the absence of tap water rinsing. METHODS: This was a descriptive, prospective microbiological study of contact lens contamination with ideal hygiene compliance and new lenses and storage cases. One hundred and fifty contact lens wearers were instructed to avoid risk factors identified for Acanthamoeba infection. They were randomly assigned to use one of three disinfecting systems and taught to follow manufacturers' instructions. In addition, they were taught to avoid all use of tap water for contact lens hygiene, except for hand washing. RESULTS: There was no isolation of Acanthamoeba from any lens storage case, precluding the chance of amoebic infection. The multi-purpose solution gave the lowest rate of bacterial contamination, with 78% sterility and 15% of cases with < 10(4) bacteria/ml. For both one-step peroxide and multi-purpose solutions, Gram-negative bacteria were reduced in frequency compared with values expected historically, while Bacillus sp. were found more frequently. Storage cases of both one-step peroxide systems leaked fluid. CONCLUSIONS: On the basis of contamination in previous studies, when hydrogen peroxide and other chemical disinfectants were used together with tap water washing, it was expected that approximately 40% of lens storage cases would yield bacteria, often with a high count, and that up to 8% would yield Acanthamoeba. Such contamination did not occur, however, in this study. The multipurpose solution, for 1 month's use, gave the lowest rate of bacterial contamination with only 7% of storage cases harbouring bacteria at > 10(4)/ml and with 78% sterility. One of the two one-step hydrogen peroxide systems performed equally well. Importantly, Acanthamoeba was not isolated from any of the 150 storage cases. Whether lens storage cases need to be sterile or contain < 10(3) bacteria/ml solution within them is debatable, but it is essential that Acanthamoeba be absent from them.

Acanthamoeba↗
PubMed1999

Fluorescent oligonucleotide probes for clinical and environmental detection of Acanthamoeba and the T4 18S rRNA gene sequence type.

The first genus- and subgenus-specific fluorescent oligonucleotide probes for in situ staining of Acanthamoeba are described. Sequences of these phylogeny-based probes complement the 18S rRNA and the gene encoding it (18S rDNA). The genus-specific probe (GSP) is a fluorescein-labeled 22-mer specific for Acanthamoeba as shown here by its hybridization to growing trophozoites of all 12 known Acanthamoeba 18S rDNA sequence types and by its failure to hybridize with amoebae of two other genera (Hartmannella vermiformis and Balamuthia mandrillaris), two human cell lines, and two bacteria (Pseudomonas aeruginosa and Escherichia coli). The sequence type T4-specific probe (ST4P) is a rhodamine-labeled 30-mer specific for Acanthamoeba 18S rDNA sequence type T4, as shown here in hybridization tests with trophozoites of all 12 sequence types. T4 is the subgenus group associated most closely with Acanthamoeba keratitis (AK). GSP also was tested with corneal scrapings from 17 patients with a high index of clinical suspicion of AK plus 5 patient controls. GSP stained both trophozoites and cysts, although nonspecific cyst wall autofluorescence also was observed. Results could be obtained with GSP in 1 to 2 days, and based on results from cell culture tests, the probe correctly detected the presence or absence of Acanthamoeba in 21 of 24 specimens from the 22 patients. The use of GSP with cultured trophozoites and cysts from corneal scrapings has illustrated the suitability of using fluorescent oligonucleotide probes for identification of the genus Acanthamoeba in both environmental and clinical samples. In addition, the use of ST4P with cultured amoebae has indicated the potential of oligonucleotide probes for use in subgenus classification.

Acanthamoeba↗
PubMed1998

Lack of evidence for systemic toxicity following topical chloramphenicol use.

There has been considerable controversy regarding the safety of topical chloramphenicol in ophthalmic practice. The evidence for associated haematopoietic toxicity in idiosyncratic and dose-dependent forms was reviewed. The 7 cases of idiosyncratic haematopoietic reactions associated with topical chloramphenicol reported in the literature are refutable evidence for the existence of such a response. In Scotland, despite extensive prescription of topical chloramphenicol, the incidence of acquired aplastic anaemia was found to be low, as were associated reports of blood dyscrasias throughout the UK. The epidemiology of acquired aplastic anaemia failed to make an association with topical chloramphenicol use. High-performance liquid chromatography (minimum detection limit 1 mg/l) was used to investigate whether serum accumulation of chloramphenicol occurred after topical therapy in 40 patients. The mean dose of chloramphenicol eye drops used after 1 week of treatment was 8.0 mg, and after 2 weeks, 15.3 mg. As expected, chloramphenicol failed to accumulate to detectable levels. This supported the view that topical chloramphenicol was not a risk factor for inducing dose-related bone marrow toxicity. Calls for the abolition of treatment with topical chloramphenicol based on current data are not supported.

Adult↗
PubMed1998

The role of Pseudomonas aeruginosa biofilm in the attachment of Acanthamoeba to four types of hydrogel contact lens materials.

PURPOSE: The effect of the previous coating of a contact lens surface with Pseudomonas biofilm on adsorption of Acanthamoeba onto four types of hydrogel materials was investigated. METHODS: Hydrogel contact lens quarters from each of the four FDA groups of hydrogel materials were incubated for at least 12 h in a suspension of 10(7)/ml of Pseudomonas aeruginosa (ATCC 27853) to coat their surfaces with biofilm. After rinsing, the lenses were incubated for 90 min in 5 x 10(5)/ml of Acanthamoeba castellanii trophozoites. New, uncoated lens quarters were incubated in the Acanthamoeba suspension as controls. After rinsing, all adsorbed trophozoites on one surface of each lens quarter were counted by direct light microscopy. Adsorption was expressed as numbers of amoebae per square centimeter of lens surface, and nonparametric data analysis was performed. RESULTS: Acanthamoeba adsorption to new, uncoated lenses was greater for ionic materials (groups 3 and 4) than for the nonionic materials (groups 1 and 2). Pseudomonas biofilm increased adsorption on all four lens types. CONCLUSION: Pseudomonas biofilm enhanced adsorption of Acanthamoeba on all lens types studied, but the adsorption to nonionic materials was significantly less. This suggests that all lens wearers may be at increased risk for Acanthamoeba infection if lenses are previously contaminated with bacterial biofilm, but this risk may be reduced by the use of certain lens types (low water content, nonionic materials).

Acanthamoeba↗
PubMed1998

Confirmatory evidence from 18S rRNA gene analysis for in vivo development of propamidine resistance in a temporal series of Acanthamoeba ocular isolates from a patient.

DNA sequences of three 18S rRNA gene alleles present in trophozoites obtained before and after therapy for Acanthamoeba keratitis substantiate a previous report that the infection was due to a single Acanthamoeba strain. Thus, the possibility that propamidine resistance which developed during therapy was due to a mixed infection was ruled out.

Acanthamoeba↗
PubMed1998

Antimicrobial management of presumed microbial keratitis: guidelines for treatment of central and peripheral ulcers.

AIMS: To determine the quantitative relation between the major risk factors for microbial keratitis of previous ocular surface disease and contact lens wear and central and peripheral infiltration, often associated with ulceration, in order to establish a rational chemotherapeutic management algorithm. METHODS: Data from 55 patients were collected over a 10 month period. All cases of presumed microbial keratitis where corneal scrapes had been subjected to microbiological examination were included. Risk factor data and laboratory outcome were recorded. Antimicrobial regimens used to treat each patient were documented. RESULTS: 57 episodes of presumed microbial keratitis were identified from 55 patients, 24 male and 31 female. There were 30 central infiltrates and 27 peripheral infiltrates of which 28 were culture positive (73% of central infiltrates, 22% of peripheral infiltrates). 26 patients had worn contact lenses of whom 12 had culture positive scrapes (9/14 for central infiltrates, 3/12 for peripheral infiltrates). 31 patients had an ocular surface disease of whom five previous herpes simplex virus keratitis patients developed secondary bacterial infection. Anterior chamber activity and an infiltrate size > or = 4 mm2 were more common with culture positive central infiltrates than peripheral infiltrates (chi 2 test = 11.98, p < 0.001). CONCLUSIONS: Predisposing factors for "presumed" microbial keratitis, either central or peripheral, were: ocular surface disease (26/57 = 45.6%), contact lens wear (26/57 = 45.6%), and previous trauma (5/57 = 8.8%). Larger ulceration (> or = 4 mm2) with inflammation was more often associated with positive culture results for central infiltration. None of these four variables (contact lens wear, ocular surface disease, ulcer size, anterior chamber activity) were of intrinsic value in predicting if a peripheral infiltrate would yield identifiable micro-organisms. Successful management of presumed microbial keratitis is aided by a logical approach to therapy, with the use of a defined algorithm of first and second line broad spectrum antimicrobials, for application at each stage of the investigative and treatment process considering central and peripheral infiltration separately.

Acanthamoeba Keratitis↗
PubMed1997

Colonization of transplant unit water supplies with Legionella and protozoa: precautions required to reduce the risk of legionellosis.

Organ transplant recipients and other immunosuppressed patients are known to be at increased risk of nosocomial Legionnaires' disease. Although the ecology of Legionella in hospital water storage and distribution systems (including a protozoonotic relationship with free-living protozoa) has been well documented, little is known regarding the quality of water supplied to high-risk units. Hot- and cold-water samples (two first draw and one run to waste for 5 min) were taken from 69 (85%) of the 81 United Kingdom organ transplant units (31 renal, 24 bone marrow, nine cardiopulmonary and five liver transplant units) and cultured for Legionella and protozoa. Legionella spp. were isolated from the water supplies of 38 (55%) units and Legionella pneumophila from 31 (45%). The blue-white fluorescent group of Legionella (Legionella gormanii, Legionella bozemanii and others) was isolated from 18 (26%) units. Free-living protozoa were isolated from 47 units (68%) and genera of the protozoa known to permit the intracellular growth of Legionella (PGIGL), from 40 units (58%). Possible associations between Legionella and the variables Protozoa; PGIGL; water pH; and circulating water temperature (recorded after running to waste for 5 min) were examined by logistic regression analysis. In cold-water supplies, a significant association was found between the isolation of Legionella and PGIGL (P = 0.032; OR = 1.81; 95% CI 1.1-3.1). In hot-water supplies, an inverse association was found between the isolation of Legionella and circulating water temperature (P = 0.034; OR = 1.0719 per degree C; 95% CI 1.0052-1.1432). (We failed to isolate Legionella when the circulating hot water was > 58 degrees C. No other associations were significant. We recommend the active surveillance of water quality in high-risk patient areas, and that transplant units, either with a history of nosocomial Legionnaires' disease, or where active surveillance indicates a persistently high Legionella colony count, take remedial action. The quality of cold water may be improved by provision of a dedicated supply taken directly from the incoming mains; and of hot water by the use of a dedicated calorifier, able to maintain a minimum circulating hot water return temperature of 60 degrees C.

Cross Infection↗