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H S Randhawa

Publications and source records attributed to H S Randhawa.

At least 19 recordsLinked to original sources

Evaluation of peptone glucose fluconazole agar as a selective medium for rapid and enhanced isolation of Aspergillus fumigatus from the respiratory tract of bronchopulmonary aspergillosis patients colonized by Candida albicans.

We have reported earlier that Aspergillus fumigatus is inhibited in vitro by Candida albicans which also interferes in its isolation from sputum experimentally seeded with predetermined graded inocula of the two fungi. It was further shown that this interference was neutralized by employing peptone glucose agar with incorporation of fluconazole which is more inhibitory to C. albicans than to A. fumigatus. This communication embodies the results of evaluation of peptone glucose fluconazole agar (PGFA) as a selective culture medium for rapid and enhanced isolation of A. fumigatus from sputum of patients clinically suspected of aspergillosis with C. albicans colonization in the respiratory tract. Of the 23 sputum specimens and one broncho-alveolar lavage collected from 15 suspected aspergillosis patients, A. fumigatus was isolated from all (100%) on PGFA as against only 19 specimens (79%) that proved to be positive on the control PGA medium (P<0.05). The greater efficacy of PGFA than that of PGA was further evident from the 2-fold higher A. fumigatus mean colony count (8.2+/-1.87) on the former medium than on the latter (3.7+/-1.00), and this difference was found to be statistically significant (P<0.05). Besides, A. fumigatus colonies were macroscopically recognizable within 2-3 days on PGFA at 28 degrees C in strong contrast to 5-7 days required on PGA. Based upon these observations, PGFA is recommended for wider application as a selective medium for rapid and enhanced recovery of A. fumigatus from sputum of patients clinically suspected of aspergillosis with C. albicans colonization in their respiratory tract.

Agar↗

Distribution of Cryptococcus gattii and Cryptococcus neoformans in decayed trunk wood of Syzygium cumini trees in north-western India.

The aim of this study is to report the regional distribution of Cryptococcus. gattii and Cryptococcus. neoformans in decayed wood inside trunk hollows of Syzygium cumini trees (Java plum, Indian black berry) investigated in Amritsar (Panjab), Meerut Cantt. and Bulandshahr (Uttar Pradesh) and Delhi, in north-western India. Two hundred and seventeen wood samples collected from 74 S. cumini trees were investigated. This includes 7 known positive S. cumini trees in Delhi subjected to a mycological surveillance for perennial colonization by C. gattii and C. neoformans. Cryptococcus gattii showed the highest prevalence (89%) in S. cumini trees in Delhi, followed by 27%, 12.5% and 9% prevalence in Bulandshahr, Amritsar City and Meerut Cantt., respectively. In contrast, C. neoformans had the highest prevalence (54%) in Amritsar, followed by 44% in Delhi, 9% in Bulandshahr and 0% in Meerut Cantt. Furthermore, 44% of the S. cumini trees in Delhi, 9% in Bulandshahr and 8% in Amritsar were concomitantly colonized by both C. gattii and C. neoformans. A mycological surveillance over 4.8-5.2 years of 7 selected S. cumini trees in Delhi revealed perennial colonization by both the Cryptococcus species. In addition, air samples taken close to the decayed trunk hollows of 4 of the perennially colonized S. cumini trees contained strains of the C. neoformans species complex. Of a random sample of 48 isolates serotyped, 26 (54%) were C. neoformans, serotype A, and 22 (46%) C. gattii, serotype B. Determination of mating type alleles was done in 44 of the isolates, comprising 31 of C. neoformans, serotype A and 13 of C.gattii, serotype B. All of them proved to be mating type alpha (MATalpha). The data on high prevalence, fungal population density, perennial colonization and aerial isolations indicate that decayed wood in trunk hollows of S. cumini trees is to-date the main well documented primary environmental niche of C. gattii and C. neoformans in north-western India. Attention is drawn to the likely health hazard posed by the environmental reservoirs of C. gattii and C. neoformans occurring in tree trunk hollows in proximity to human and animal habitations.

Cryptococcus neoformans↗

Malassezia furfur in a case of onychomycosis: colonizer or etiologic agent?

The etiologic role of Malassezia furfur in onychomycosis is a contentious diagnostic problem because its keratinolytic ability has never been verified. This case report describes the isolation of M. furfur from the infected nails of a child clinically diagnosed with onychomycosis, and discusses the role of this organism as an etiologic agent/colonizer. The patient presented with subungual hyperkeratosis and onycholysis without associated paronychia. Budding yeast cells compatible with M. furfur were repeatedly demonstrated in KOH wet mounts of damaged nails, histopathology of hematoxylin and eosin (H&E) and periodic acid-Schiff (PAS) stained sections showed penetration of fungal elements between deeper layers of keratin, and numerous colonies of M. furfur were isolated on three consecutive occasions from nail specimens collected from different areas of hand and toenail lesions. No evidence of nail invasion by dermatophytic or nondermatophytic filamentous fungi were found by direct microscopy or culture. Microscopy and culture were negative following 12 weeks of ketoconazole treatment, which resulted in growth of healthy nail plates with normal beds. We can infer from these observations that M.furfur was an etiologic agent rather than a colonizer in the patient's nails even though direct keratinolytic character of this fungus was not demonstrated.

Adolescent↗

Efficacy of swabbing versus a conventional technique for isolation of Cryptococcus neoformans from decayed wood in tree trunk hollows.

The efficacy of swabbing versus a conventional sedimentation technique was evaluated for sampling of decayed wood in tree trunk hollows for isolation of Cryptococcus neoformans. Of 52 samples of decayed wood, bark or other plant debris originating from 35 living trees, 42 wood samples yielded C. neoformans. The positive samples included 40 collected from 31 Syzygium cumini trees growing along roadsides in Old Delhi, whereas the remaining two were from inside tree trunk fissures of Ficus religiosa in a New Delhi locality. The number of wood samples found positive by swabbing was 40 (95%) as opposed to 32 (76%) by the conventional technique, and this difference was statistically significant (P < 0.01). Also, the conventional technique showed 24% false-negative results, which was in striking contrast to only 5% by swabbing. Furthermore, swabbing yielded a significantly higher C. neoformans mean colony count than did the conventional technique (P < 0.005), thus highlighting greater efficacy of the former technique. The overall prevalence of C. neoformans in the S. cumini trees investigated was 84% (26/31 trees) which is the highest as yet reported from any tree species in India. Varietal identification and serotyping was done with 33 of the C. neoformans isolates, 31 of which came from 23 tree trunk hollows of S. cumini and two from the tree trunk fissures of F. religiosa. Among the S. cumini isolates, 26 were identified as C. neoformans var. gattii (all serotype B except two untypeable ones) and five as C. neoformans var. neoformans, serotype A (= C. neoformans var. grubii). Both of the F. religiosa isolates belonged to C. n. var. neoformans, serotype A. Being a more efficacious, simple, less time-consuming and less hazardous technique, swabbing is recommended for wider use in order to further elucidate the ecology of C. neoformans.

Biodegradation, Environmental↗

Development of an expressed sequence tag (EST) resource for wheat (Triticum aestivum L.): EST generation, unigene analysis, probe selection and bioinformatics for a 16,000-locus bin-delineated map.

This report describes the rationale, approaches, organization, and resource development leading to a large-scale deletion bin map of the hexaploid (2n = 6x = 42) wheat genome (Triticum aestivum L.). Accompanying reports in this issue detail results from chromosome bin-mapping of expressed sequence tags (ESTs) representing genes onto the seven homoeologous chromosome groups and a global analysis of the entire mapped wheat EST data set. Among the resources developed were the first extensive public wheat EST collection (113,220 ESTs). Described are protocols for sequencing, sequence processing, EST nomenclature, and the assembly of ESTs into contigs. These contigs plus singletons (unassembled ESTs) were used for selection of distinct sequence motif unigenes. Selected ESTs were rearrayed, validated by 5' and 3' sequencing, and amplified for probing a series of wheat aneuploid and deletion stocks. Images and data for all Southern hybridizations were deposited in databases and were used by the coordinators for each of the seven homoeologous chromosome groups to validate the mapping results. Results from this project have established the foundation for future developments in wheat genomics.

Blotting, Southern↗

Deletion mapping of homoeologous group 6-specific wheat expressed sequence tags.

To localize wheat (Triticum aestivum L.) ESTs on chromosomes, 882 homoeologous group 6-specific ESTs were identified by physically mapping 7965 singletons from 37 cDNA libraries on 146 chromosome, arm, and sub-arm aneuploid and deletion stocks. The 882 ESTs were physically mapped to 25 regions (bins) flanked by 23 deletion breakpoints. Of the 5154 restriction fragments detected by 882 ESTs, 2043 (loci) were localized to group 6 chromosomes and 806 were mapped on other chromosome groups. The number of loci mapped was greatest on chromosome 6B and least on 6D. The 264 ESTs that detected orthologous loci on all three homoeologs using one restriction enzyme were used to construct a consensus physical map. The physical distribution of ESTs was uneven on chromosomes with a tendency toward higher densities in the distal halves of chromosome arms. About 43% of the wheat group 6 ESTs identified rice homologs upon comparisons of genome sequences. Fifty-eight percent of these ESTs were present on rice chromosome 2 and the remaining were on other rice chromosomes. Even within the group 6 bins, rice chromosomal blocks identified by 1-6 wheat ESTs were homologous to up to 11 rice chromosomes. These rice-block contigs were used to resolve the order of wheat ESTs within each bin.

Chromosome Mapping↗

A chromosome bin map of 16,000 expressed sequence tag loci and distribution of genes among the three genomes of polyploid wheat.

Because of the huge size of the common wheat (Triticum aestivum L., 2n = 6x = 42, AABBDD) genome of 17,300 Mb, sequencing and mapping of the expressed portion is a logical first step for gene discovery. Here we report mapping of 7104 expressed sequence tag (EST) unigenes by Southern hybridization into a chromosome bin map using a set of wheat aneuploids and deletion stocks. Each EST detected a mean of 4.8 restriction fragments and 2.8 loci. More loci were mapped in the B genome (5774) than in the A (5173) or D (5146) genomes. The EST density was significantly higher for the D genome than for the A or B. In general, EST density increased relative to the physical distance from the centromere. The majority of EST-dense regions are in the distal parts of chromosomes. Most of the agronomically important genes are located in EST-dense regions. The chromosome bin map of ESTs is a unique resource for SNP analysis, comparative mapping, structural and functional analysis, and polyploid evolution, as well as providing a framework for constructing a sequence-ready, BAC-contig map of the wheat genome.

Chromosome Mapping↗

Decayed wood of Syzygium cumini and Ficus religiosa living trees in Delhi/New Delhi metropolitan area as natural habitat of Cryptococcus neoformans.

The isolation is reported of Cryptococcus neoformans var. gattii and C. n. var. neoformans from decayed wood inside trunk hollows of Syzygium cumini and of C. n. var. neoformans from Ficus religiosa trees in the Delhi/New Delhi metropolitan area. Fourteen of sixty-six (21%) S. cumini trees investigated proved to be positive, seven for each variety. The two varieties never co-occurred in the same hollow. C. n. var. neoformans was also isolated from three of seventeen Ficus religiosa-trees. Two of these isolates originated from decayed wood and one from bark. The C. n. var. gattii and C. n. var. neoformans isolates belonged to serotype B and serotype A, respectively. The data strongly supported colonization of S. cumini by both varieties and of F. religiosa trees by C. n. var. neoformans. Evidence of this was found by repeated isolations. For example, in 36/44 (82%) samples for C. n. var. gattii and 22/27 (81%) samples for C. n. var. neoformans, and by a high population density in the tested wood debris (maximally 6 x 10(5) colony-forming units per gram [c.f.u./g] for C. n. var. gattii and 8 x 10(4) c.f.u./g for C. n. var. neoformans). No eucalypt trees were seen near the positive S. cumini and F. religiosa trees. The densities of C. neoformans in these trees exceeded those found previously in Eucalyptus camaldulensis and in other tree species more rarely reported to be sources of C. neoformans in India. S. cumini and F. religiosa appear not to have been reported to date as sources for either C. n. var. gattii or C n. var. neoformans. Our results add to the recently emerging evidence that the natural habitat of C. n. var. gattii and C. n. var. neoformans is not specific to woody or other debris of particular tree species, but instead is more generalized.

Colony Count, Microbial↗

Decaying wood in tree trunk hollows as a natural substrate for Cryptococcus neoformans and other yeast-like fungi of clinical interest.

The occurrence of Cryptococcus neoformans var. neoformans and other yeast-like fungi of clinical interest in decaying wood inside tree trunk hollows, bark and other plant materials is reported. The var. neoformans was isolated from 3 of 45 (6.6%) wood and one of 390 Eucalyptus bark samples. Two of the positive wood samples came from a tree trunk hollow of Butea monosperma (Family: Papilionaceae) growing in Roshan Ara Garden, Old Delhi whereas the third was from a trunk hollow of Tamarindus indica (Family: Papilionaceae) growing outside of Talkatora Garden, New Delhi. The solitary positive Eucalyptus bark sample originated from Amritsar. The isolations of var. neoformans from decaying wood inside trunk hollows of B. monosperma and T indica constitute the first record of the natural occurrence of this pathogen in association with these trees. The observation reinforces the recent evidence for decaying wood inside trunk hollows of some trees to be a new natural habitat of the variety neoformans. Besides, in consonance with their essentially saprobic character, a number of other yeast-like fungi were sporadically isolated. This includes, Cryptoccus laurentii, Cryptococcus albidus, Candida lusitaniae, C. guilliermondii, C. krusei, C. tropicalis, C. zeylanoides, Trichosporon cutaneum, Rhodotorula mucilaginosa, R. glutinis, Geotrichum capitatum, G. klebahnii and Sporobolomyces salmonicolor. Cryptococcus neoformans var. gattii was not found in any of the 702 samples of plant materials, including the bark and detritus of Eucalyptus camaldulensis and E. tereticornis trees. A more extensive environmental survey, covering divergent climatic regions, is warranted to identify the natural reservoirs of var. gattii in India.

Cryptococcus neoformans↗

Thermophilic actinomycetes in cane sugar mills: an aeromicrobiologic and seroepidemiologic study.

Aerial prevalence of clinically important thermophilic actinomycetes and occurrence of precipitating antibodies against them in sera of 153 exposed workers have been reported. The study was carried out in two cane sugar mills namely, the Upper Doab Sugar Mills and the Ramala Sugar Mills, located in north-west India. In both the sugar mills, T. sacchari was the predominant species, it accounted for 55.1% and 50.3% of the total population of thermophilic actinomycetes, followed by T. vulgaris (19.7% and 23.7%), T. thalpophilus (21.1% and 17.1%), Saccharomonospora viridis (3.4% and 5.0%) and Saccharopolyspora rectivirgula (Faenia rectivirgula) (0.7% and 3.9%), respectively. Precipitating antibodies against thermophilic actinomycetes were demonstrable in 34 (22.2%) workers; T. sacchari alone accounted for 20 of the positive precipitin reactions, followed by S. rectivirgula in 10. The mean absorbance values for IgG antibody activity against T. sacchari as well as S. rectivirgula were found to be elevated significantly in the symptomatic workers than in the asymptomatic workers (p < 0.05) or unexposed controls (p < 0.001). However, the difference in IgG antibody activity was insignificant between precipitin-positive symptomatic workers and precipitin-positive asymptomatic workers. The results indicate that clinically important thermophilic actinomycetes are widely prevalent in cane sugar mills, and T. sacchari and S. rectivirgula are the major species involved in the sensitization of the bagasse workers in India.

Actinomycetales↗

Epididymal histoplasmosis diagnosed by isolation of Histoplasma capsulatum from semen.

An autochthonous case of epididymal histoplasmosis masquerading as tuberculosis in a 55-year-old male patient is reported from India. It was diagnosed by culture of Histoplasma capsulatum from semen and by demonstration of the fungus upon re-examination of epididymal biopsy sections previously misinterpreted as tuberculous granuloma. The patient's main complaints were painful epididymal swelling, occasional fever and cough. He was treated successfully by excision of epididymis and vas deferens combined with amphotericin B therapy. This is believed to be the first case of epididymal histoplasmosis to be reported outside the American continent and the fourth of its type reported in the English literature. The case is also noteworthy in that H. capsulatum was isolated for the first time from semen, and it underlines the importance of mycological culture of semen specimens for diagnosis of genitourinary infections of obscure etiology.

Biopsy↗

Nomenclature of fungal diseases: a report and recommendations from a Sub-Committee of the International Society for Human and Animal Mycology (ISHAM).

The ISHAM Mycoses Nomenclature Committee has considered the present status of fungal disease names. It suggests that the traditional approach to mycoses nomenclature in which the name of a causative taxon is suffixed with '-asis', '-iasis', '-osis' or '-mycosis' leads to names that are frequently unstable with respect to subsequent taxonomic and clinico-epidemiological changes. It is therefore recommended that individual mycoses should be named as often as possible in the form 'pathology A due to/caused by fungus X' or '[adjectival] fungus X pathology A' in preference to construction of names based solely on fungal taxa. A list of recommended mycosis names retained for their long tradition or intrinsic convenience is provided, together with a combined index and list of rejected names.

Animals↗

Efficacy of seed-based media for the mould-yeast conversion of Blastomyces dermatitidis.

The efficacy of 20 seed-based media is reported for the in vitro mould-yeast conversion of Blastomyces dermatitidis, employing pharmamedia agar, peptone glucose agar, glucose agar and water agar as controls. The mould-yeast conversion varied significantly according to the culture medium, fungal strains and incubation period (p less than 0.01). Garden-pea, chick-pea, cow-pea, soybean, peanut, green gram, French bean, lentil, okra and cottonseed converted all of the 7 B. dermatitidis test strains after 5 days of incubation at 37 degrees C. Although the efficacy of many of these seed media was found to be at par with pharmamedia agar - a commercial cottonseed embryo-derived protein, garden-pea seed agar is adopted because of the wider availability and low fat content of this seed. The recommended composition of the medium comprises 2% aqueous seed extract, 2% glucose and pH 6-7. Only nigerseed and sunflower seeds failed to support the conversion of B. dermatitidis. Of the control media, peptone glucose agar, glucose agar and water agar did not support the conversion of 2 of the B. dermatitidis test strains. The mechanism underlying variable mould-yeast conversion of B. dermatitidis on seed-based media is not clearly understood. However, most of the seeds supporting excellent mould-yeast conversion are known for their high protein content. The conversion was apparently not affected by the fat content of the seeds or by incorporation of glucose in the medium.

Blastomyces↗

Occurrence and significance of precipitating antibodies against thermophilic actinomycetes in the sera of dairy herd workers, Nangali, Delhi.

The study was prompted by the lack of information on the role of thermophilic actinomycetes in hypersensitivity pneumonitis in India. It reports the occurrence of precipitating antibodies against clinically important thermophilic actinomycetes in the sera of a population sample of dairy herd workers, Nangali, Delhi. Of 112 workers investigated, 28 (25%) showed precipitins against Faenia rectivirgula, 4 (3.2%) against Saccharomonospora viridis, 2 against Thermoactinomyces thalpophilus and one each against T. vulgaris and T. sacchari. The results of enzyme-linked immunosorbent assay (ELISA) indicated that IgG antibody activity against F. rectivirgula was significantly higher in the symptomatic group than in the asymptomatic group (p less than 0.05) of workers and the controls (p less than 0.01). Significant difference in F. rectivirgula IgG activity was also obtained between the precipitin-positive symptomatic group and the precipitin-positive asymptomatic group (p less than 0.05). In strong contrast, the IgG antibody activity against T. thalpophilus was found to be uniformly low. A limited aeromicrobiological sampling of the dairy farm revealed S. viridis (55.8%) to be the commonest species followed by T. vulgaris (19.2%), T. thalpophilus (18.5%), F. rectivirgula (5%) and T. sacchari (1.5%). On the basis of suggestive clinical and laboratory findings, farmer's lung disease was suspected in four dairy herd workers. A comprehensive clinical evaluation including pulmonary function studies on the dairy herd workers and their long-term follow-up is indicated to determine the extent of respiratory morbidity caused by F. rectivirgula, S. viridis, T. thalpophilus, T. sacchari and T. vulgaris in India.

Adult↗

Efficacy of brain heart infusion-egg albumen agar, yeast extract phosphate agar and peptone glucose agar media for isolation of Blastomyces dermatitidis from sputum.

The efficacy of brain heart infusion (BHI)-egg albumen agar, yeast extract phosphate agar and several modified peptone glucose agar media was evaluated for isolation of Blastomyces dermatitidis from sputum concomitantly seeded with the yeast form of the pathogen and Candida albicans. Based upon high per cent culture positivity of sputum, improved recovery (CFU/ml) of the seeded inoculum, faster growth rate of B. dermatitidis and low level of contamination, BHI-egg albumen agar, followed by yeast extract phosphate agar are recommended as the media of choice for the isolation of B. dermatitidis from contaminated clinical specimens.

Blastomyces↗

Isolation and partial characterization of a plasma membrane-enriched fraction of Blastomyces dermatitidis.

The isolation and characterization of a plasma membrane-enriched fraction is reported from the pathogenic fungus Blastomyces dermatitidis employing a simplified and inexpensive procedure. The yeast cells, suspended in an osmotic stabilizer buffer, were subjected to gentle mechanical disruption followed by a three-step differential centrifugation to obtain a 40,000 x g sediment. This fraction showed 2.7- and 3.3-fold relative enrichment of the plasma membrane marker enzymes Mg2(+)-ATPase and 5'-nucleotidase, respectively, with modest cross-contamination of other intracellular membranes. The fraction comprised approximately 49% protein, 31% carbohydrate, 11% phospholipids and 9% sterol with sterol to phospholipid molar ratio of 1.1. Transmission electron microscopy of the plasma membrane-enriched fraction showed homogeneous vesicles with a tripartite unit membrane of 115 A thickness.

5'-Nucleotidase↗