Hair Baiting Technique Research Articles

Prevalence of keratinophilic fungi in public park soils of Mumbai, India

The parks of Mumbai are frequently visited by local residents every morning and evening. However, there are no reports on the occurrence of keratinophilic fungi in these areas. The purpose of this research was to study the occurrence of keratinophilic fungi in the public parks of Mumbai. One hundred soil samples were collected from five public parks: Kamla Nehru Park, Powai Garden, CD Deshmukh Garden, Five Gardens and Chota Kashmir. Keratinophilic fungi were isolated by the hair baiting technique using human hair as keratin bait. The cultures were identified using macroand micro-morphological features. Identification was also confirmed by the BLAST search of sequences of the ITS1-5.8S-ITS2 rDNA region against the NCBI/Genbank data and compared with deposited sequences. The ability of these fungi to use human hair was also evaluated by release of protein in liquid media. A total of 75 strains of keratinophilic fungi were recovered from 100 (75.0%) soil samples. The isolated fungi were composed of eleven species of eight genera: <em>Arthrographis kalrae</em>, A<em>uxarthron conjugatum</em>, <em>Chrysosporium indicum</em>, <em>C. queenslandicum</em>, <em>C. zonatum, Gymnascella dankaliensis</em>, <em>G. hyalinospora</em>,<em> Microsporum gypseum</em> (15.0%), <em>Myriodontium keratinophilum</em>, <em>Trichophyton mentagrophytes</em> and <em>Uncinocarpus reesii</em>. These fungi can release 148.8-307.6 μg/mL protein in liquid media when grown on human hair in shake flask culture and also decompose 16.2-38.6% of human hair after four weeks of incubation. Our study indicates that keratinophilic fungi are to be found in the soils of various public parks in Mumbai and that human hair can be a source of pathogenic fungi.

Isolation of Microsporum gypseum in soil samples from different geographical regions of Brazil, evaluation of the extracellular proteolytic enzymes activities (keratinase and elastase) and molecular sequencing of selected strains

A survey of Microsporum gypseum was conducted in soil samples in different geographical regions of Brazil. The isolation of dermatophyte from soil samples was performed by hair baiting technique and the species were identified by morphology studies. We analyzed 692 soil samples and the recuperating rate was 19.2%. The activities of keratinase and elastase were quantitatively performed in 138 samples. The sequencing of the ITS region of rDNA was performed in representatives samples. M. gypseum isolates showed significant quantitative differences in the expression of both keratinase and elastase, but no significant correlation was observed between these enzymes. The sequencing of the representative samples revealed the presence of two teleomorphic species of M. gypseum (Arthroderma gypseum and A. incurvatum). The enzymatic activities may play an important role in the pathogenicity and a probable adaptation of this fungus to the animal parasitism. Using the phenotypical and molecular analysis, the Microsporum identification and their teleomorphic states will provide a useful and reliable identification system.

Prevalence of keratinophilic fungi in usar soils of Uttar Pradesh, India

Alkaline soils commonly called as <em>Usar</em> soil having pH 7.5 to 11.0 is commonly found in regions having poor drainage and little percolation. These soils occupy vast tracts of barren lands in the northern parts of India. These areas are frequently visited by man and animals, however there are no reports on the occurrence of keratinophilic fungi from the <em>Usar</em> soils. The purpose of this research was to study the occurrence of keratinophilic fungi in the alkaline Usar soil. To investigate the occurrence of soil keratinophilic fungi from <em>Usar</em> soil, 120 samples were collected from five districts of Uttar Pradesh viz. Lucknow, Bareilly, Azamgarh, Balia and Pratapgarh. Keratinophilic fungi were isolated by the hair baiting techniques using human hair as keratin bait. The cultures were identified using macro- and micro morphological features. Eighty-tow strains of keratinophilic fungi were recovered from 120 (68.33%) soil samples. The isolated fungi were composed of ten species of five genera viz. <em>Chrysosporium indicum</em> (19.16%), <em>Microsporum gypseum</em> (12.5%), <em>Chrysosporium tropicum</em> (10.0%), <em>Chrysosporium pannicola</em> (7.5%), <em>Trichophyton terrestre</em> (5.83%), <em>Chrysosporium lucknowense</em> (4.16%), <em>Chrysosporium</em> state of <em>Ctenomyces serratus</em> (3.33%), <em>Gymnascella dankailensis</em> (3.33%), <em>Gymnoascus reessii</em> (1.66 %) and <em>Trichophyton mentagrophytes</em> (1.66 %). The presence of keratinophilic fungi in the <em>Usar</em> soils of Uttar Pradesh, India indicates that these fungi do occur at higher alkalinity.

Isolation of soil dermatophytes from three distinct geographic locations in Jordan

Abstract Due to the unique geography of Jordan, which comprises mountains, valleys and desert terrains, the isolation of dermatophytes from soils derived from these three distinct regions was investigated. Ten soil samples were collected from each terrain and investigated using the hair bait technique. Standard procedures were employed to identify the isolates. Results demonstrated that desert soil was devoid of dermatophytes, whereas nine and six samples from the mountains and valleys respectively were positive. A total of 42 isolates were identified and they belonged to six species. All six species were recovered from the mountain soils and only four were detected in soils from the valleys. With the exception of Trichophyton mentagrophytes , which is a zoophilic fungus, all isolates were geophilic. The results indicated a low diversity of dermatophytes in the geographic regions studied and suggested that the isolates were unlikely to cause human dermatophytoses.

Keratinophilic fungi in activated sludge of wastewater treatment plants with MBR in Berlin, Germany

Seventy-two samples of aerobic and anoxic activated sludge were collected from two wastewater treatment plants (36 from each) with membrane bioreactors (MBR) in Berlin (Germany) to determine the presence of cycloheximide-resistant, keratinophilic fungi using the hair-baiting technique. This is the first study on the occurrence of these fungi in MBR sludges. Forty-six fungal species belonging to 21 genera were isolated and identified (41 species and 21 genera versus 36 species and 19 genera from aerobic and anoxic sludge samples, respectively). Twelve species were dermatophytes or closely related fungi: Chrysosporium asperatum, C. georgii, C. indicum, C. keratinophilum, C. tropicum, C. pannorum, Microsporum cookei, M. ferrugineum, M. gypseum, Trichophyton ajelloi, T. mentagrophytes and T. terrestre. The other 32 species, included membe rs of Acremonium, Alternaria, Aspergillus, Candida, Chaetomium, Cladosporium, Fusarium, Geotrichum, Gliocladium, Gymnoascus, Mucor, Paecilomyces, Penicillium, Scopulariopsis, Sporothrix, Trichoderma, Verticillium and unidentified yeasts. The fungi isolated are pathogenic or potentially pathogenic to humans and animals, and their occurrence in activated sludge poses a health risk to wastewater treatment workers and the environment.

Carbohydrate profiling of fungal cell wall surface glycoconjugates of Trichophyton tonsurans and other keratinophilic filamentous fungi using lectins

Various researchers have concluded that lectins are useful reagents for the study of fungal cell wall surface glycoconjugates. In this study, we evaluated the expression of N-acetyl-D-glucosamine, L-fucose, D-galactose and glucose/mannose on the cell wall surface of Trichophyton tonsurans and other keratinophilic filamentous fungi, using a simple lectin-binding protocol. The fungal cultures used were isolated from soils obtained from public parks by the hair-bait technique. The lectin assays used concanavalin A (Con A), wheat germ agglutinin (WGA), Ulex europeus agglutinin I (UEA-I) and peanut agglutinin (PNA), all conjugated with horseradish peroxidase. Adhesive tape was placed sticky-side down over the fungal colony, gently pressed and then removed. The fungal-tape samples were incubated with the lectin for 1 h at 4 °C. Lectin binding was visualised using 3,3-diaminobendizine (DAB) and hydrogen peroxidase. There was a high expression of N-acetyl-D-glucosamine on the cell wall surface of all fungi species tested, whereas the expression of L-fucose, D-galactose and glucose/mannose demonstrated inter-specific variations. The lectin-binding assay presented in this article eliminates many of the laborious steps involved in other protocols. The amount and quality of the mycelium and spores immobilised by the adhesive tapes were suitable for obtaining the carbohydrate profile in glycoconjugates of the cell wall surface of filamentous fungi.

Production of Antibacterial Like Substances from Some Clinical Isolates of Trichophytic Genera of Dermatophytes

Nine clinical and six soil isolates of Trichophytic genera of dermatophytes were demonstrated for their in vitro production of antibacterial substances. Clinical isolates were isolated from superficial skin of patients suffering from dermatomycoses and soil isolates were procured from soil using hair-baiting technique. Among nine clinical isolates of dermatophytes tested for their ability to produce antibacterial substances, five were found to be producers and among six soil isolates, three produces the antibacterial substances. The qualitative identification of the produced antibiotics reveals five types; a penicillin-like substance produced by Trichophyton concentricum, and two different types of unknown substances obtained from Trichophyton mentagrophytes and T megninii, the kojic acid-like antibacterial substance produced by Trichophyton verrucosum. The fusidic acid like substances was obtained from T tonsurans and T simii.

Incidence of keratinophilic fungi from the soils of Vedanthangal Water Bird Sanctuary (India)

Fifty-three soil samples were collected from various sites in the vicinity of Vedanthangal Water Bird Sanctuary and screened for the presence of keratinophilic fungi using the hair baiting techniques for isolation. Twenty-eight isolates were recovered and identified by recognition of their macro- and micromorphological features. Seven species related to five genera were recorded viz. Auxarthron conjugatum (1.89%), Chrysosporium fluviale (3.77%), Chrysosporium indicum (20.75%), Chrysosporium tropicum (7.55%), Chrysosporium state of Ctenomyces serratus (5.66%), Gymnoascus petalosporus (1.89%) and Microsporum gypseum complex (11.32%). The study shows that migratory birds harbour a variety of keratinophiles and may be a potential source of transfer of these fungi from one location to another.

The occurrence of keratinophilic fungi in selected soils of Ladakh (India)

138 soil samples were collected from various loca-tions in Ladakh, a cold desert in the Himalayan region, India and the samples were screened for the presence of keratinophilic fungi using the hair baiting techniques. 58 isolates were recovered and identified. The cultures were identified based on their macro- and micro-morphological features. A total of six genera and fourteen species were isolated namely Amauroascus kuehnii (0.72%), Aphanoascus keratinophilus (4.34%), Aphanoascus terreus (2.17%), Auxarthron alboluteum (0.72%), Auxarthron conjugatum (0.72%), Chrysosporium articulatum (0.72%), Chrysosporium mephiticum (0.72%), Chrysosporium minutisporosum (2.17%), Chrysosporium siglerae (0.72%), Chrysosporium sp. (1.44%), Chrysosporium tropicum (15.94 %), Chrysosporium submersum (3.62%), Chrysosporium state of Ctenomyces serratus (6.52%) and Geomyces pannorum (1.45%). The present study shows that keratinophilic fungi exist in the cold desert of Ladakh.

Keratinophilic fungi in soil of potted plants of indoor environments in Kanpur, India, and their proteolytic ability

One hundred nine isolates of 10 genera representing 20 species of keratinophilic fungi were isolated from soil of planted earthen pots in indoor environments of 15 localities of Kanpur, India, by the hair baiting technique. Microsporum gypseum, Trichophyton vanbreuseghemii, and Botryotrichum piluliferum were found to release 698.66, 512.99, and 519.99μg/ml net protein, respectively, whereas Arthroderma cuniculi released the minimum, 107.99μg/ml. All other isolates were keratinolytically active.

DERMATOPHYTES, RELATED KERATINOPHILIC AND OPPORTUNISTIC FUNGI IN INDOOR DUST OF HOUSES AND HOSPITALS

Dermatophytes, related keratinophilic and opportunistic fungi were isolated from indoor dust samples of 46 hospitals and 47 houses in Kanpur. A total of 19 fungi represented by 11 genera were isolated by the hair-baiting technique from 230 and 235 samples from hospitals and houses respectively. The isolated fungi are Acremonium implicatum (Indian Type Culture Collection) ITCC 5266, A. strictum (Germplasm Centre for Keratinophilic Fungi) GPCK 1137, Aphanoascus fulvescens GPCK 1081, Arthroderma simii GPCK 1275, Chrysosporium queenslandicum ITCC 5270, C. indicum ITCC 5269, C. pannicola GPCK 1022, C. tropicum GPCK 1269, Ctenomyces serratus ITCC 5267, Gymnoascus reessii ITCC 5265, Malbranchea fulva GPCK 1075, Malbranchea pulchella ITCC 5268, Micosporum gypseum GPCK 1038, Microsporum cookei GPCK 2001, M. fulvum GPCK 2002, Paecilomyces lilacinum GPCK 1080, Penicillium expansum GPCK 1082, Trichophyton mentagrophytes GPCK 2003 and T. terrestre GPCK 2004. In hospitals, the minimum frequency was of Ctenomyces serratus ITCC 5267 while the maximum frequency was of Arthroderma simii GPCK 1275. In houses, Chrysosporium queenslandicum ITCC 5270 and C. tropicum GPCK 1269 were with minimum and maximum frequencies respectively. This makes the first report of these fungi with keratinolytic ability in the indoor dust of hospitals and houses.

P247 Fertility and viability rates of hydatid cysts in slaughtered animals in the Mazandaran Province, Northern Iran

A total of 23 soil samples of potted plants was collected from hospitals in Sari, Iran. Each sample contained approximately 200 g soil, taken from a depth of 0–10 cm of the pots. Samples were analysed by two different methods. (1) Culture on Sabouraud's dextrose agar medium containing chloramphenicol (SC); cultured fungi were identified by macroscopic and microscopic characterization. (2) The hair-baiting technique (HBT) was used to isolate the keratinophilic fungi. After culture on SC medium, 100% of plates were positive for fungal growth, and in total 1150 colonies with 13 different types of fungi were isolated. Penicillium spp. (52%), Acremonium spp. (20%), Paecilomyces spp. (11.9%), Cladsporium spp. (3.7%) and Aspergillus spp. (3.1%) were the predominant fungal species. Rhizopus spp. (0.1%) were less frequent. Keratinophilic fungi such as Microsporum gypseum, Microsporum cookei and Chrysosporium spp. were isolated using HBT. The presence of pathogenic fungi such as Cladosporium spp, Aspergillus spp, M. gypseum, and M. cookei in potted plants in hospitals represents a potential source of nosocomial infection.

Survey of keratinophilic fungi in sewage sludge from wastewater treatment plants of Mazandaran, Islamic Republic of Iran

To isolate keratinophilic fungi in sewage sludge from wastewater treatment plants in Sari city, Mazandaran province, Islamic Republic of Iran, samples were taken from 7 plants with different sewage treatment technologies. From 35 sludge samples cultured on Sabouraud's agar with cycloheximide and chloramphenicol, 326 fungal colonies belonging to 7 species were isolated. Geotrichum (59.5%), Cladosporium (13.8%), Alternaria (11.3%) and Penicillium (10.7%) species were the most prevalent. No growth of keratinophilic fungi was observed on this medium. However, using the hair-baiting technique, Microsporum gypseum, Chrysosporium spp. and Geotrichum spp. were isolated.

Keratinophilic fungi in soil of potted plants of indoor environments in Kanpur, India, and their proteolytic ability

One hundred nine isolates of 10 genera representing 20 species of keratinophilic fungi were isolated from soil of planted earthen pots in indoor environments of 15 localities of Kanpur, India, by the hair baiting technique. Microsporum gypseum, Trichophyton vanbreuseghemii, and Botryotrichum piluliferum were found to release 698.66, 512.99, and 519.99μg/ml net protein, respectively, whereas Arthroderma cuniculi released the minimum, 107.99μg/ml. All other isolates were keratinolytically active.

Isolation and identification of the human pathogenPythium insidiosumfrom environmental samples collected in Thai agricultural areas

We describe the ecological niche of the human and animal pathogen Pythium insidiosum within endemic agricultural areas of Thailand. Samples were collected from irrigation water, including rice paddy fields, irrigation channels and reservoirs. Zoospores of P. insidiosum were captured from water by the use of a sterile human hair baiting technique. Pythium isolates were identified based on phenotypic characteristics and by using a specific PCR assay for P. insidiosum. In addition, internal transcribed spacer (ITS) regions of P. insidiosum rDNA were sequenced and used in the phylogenetic analysis of 20 other known P. insidiosum DNA sequences available in the database and 11 related DNA sequences of other Pythium species including Lagenidium giganteum. The sequences of 59 environmental isolates of Pythium spp. recovered from Thailand confirmed 99% identity to P. insidiosum. Three well supported phylogenetic groups within P. insidiosum were found. The protein profiles of P. insidiosum environmental strains were determined and compared with reference strains. A typical 45-30 kDa band was consistently found in all isolates of P. insidiosum but not in closely related Pythium species. This study provides the first evidence for the natural occurrence of P. insidiosum in endemic aquatic environments. The highest recovery rate of this hydrophilic pathogen was found to be from water reservoirs and our data show that irrigation water may be an important source of P. insidiosum infection for individuals working in endemic agricultural areas.

Keratinophilic fungi from selected soils of Bahrain

Eighty soil samples were collected from various sites of Bahrain and screened for presence of keratinophilic fungi using hair baiting techniques for isolation. Thirty-six isolates were recovered and identified. The cultures were identified using macro- and micromorphological features. Their identification was also confirmed by the BLAST search of sequences of ITS1-5.8S-ITS2 rDNA region against the NCBI/Gene bank data and compared with deposited sequences for confirmation. Eight species of five genera were isolated viz. Aphanoascus fulvuscence (8.75%), Aphanoascus punsolae (20.00%), Chrysosporium indicum (2.50%), Chrysosporium tropicum (2.50%), Chrysosporium zonatum (3.75%), Spiromastix warcupii (1.25%), Microsporum gypseum (3.75%), and Trichophyton mentagrophytes (2.50%). In conclusion, our study indicates that keratinophilic fungi do occur in the various soils of Bahrain. Moreover, the narrow diversity and low density of keratinophilic fungi in the investigated soils is expected and is emblematic to other hot arid environments.

Keratinophilic fungi from the vicinity of meteorite crater soils of Lonar (India)

Thirty-two soil samples were collected from six sites in the vicinity of Lonar lake, a meteorite crater and screened for presence of keratinophilic fungi using hair baiting techniques for isolation. Seventeen isolates were recovered and identified. The cultures were identified by recognition of their macro- and micromorphological features. Their identification was also confirmed by the BLAST search of sequences of ITS1-5.8S-ITS2 rDNA region against the NCBI/Gene bank data and compared with deposited sequences for identification purpose. Seven species of four genera were isolated viz. Aphanoascus durus (9.38%), Aphanoascus punsolae (3.125%), Auxarthron kuehnii (3.125%), Chrysosporium indicum (25.00%), Chrysosporium tropicum (3.125%), Chrysosporium sp. (3.125%), Chrysosporium state of Ctenomyces serratus (6.25%).

SEASONAL BEHAVIOUR OF KERATINOPHILIC FUNGI ISOLATED FROM PARKS SOIL IN MILAN

Soil samples collected from 10 parks in Milan were examined for the presence of keratinophilic fungi by the Orr’s hair baiting technique. The organisms isolated and their frequency were as follows: Arthroderma uncinatum and its anamorph Keratinomyces ajelloi were dominant and recovered from 22.5% - 43.7% and 40% - 50% of the soil samples collected in summer and winter respectively. Myceliophthora vellerea was isolated in 31.2% of the samples, particularly in winter, Microsporum gypseum in winter (20%) and Trichophyton terrestre (10%) in summer. Chrysosporium keratinophilum (16.2% in summer and 18.7% in winter) C. indicum (15% s- 23.7% w), C. pannicola (5% s - 37% w). Aphanoascus fulvescens was isolated in summer (23.7%). Among the cycloheximide-resistant keratinophilic fungi Alternaria alternata (27.5% s – 33.7% w), Paecilomyces lilacinus (26.2% s - 18.7% w) and Acremonium strictum (8.7% s - 20% w) also were observed. The distribution of the different species is discussed.

Keratinophilic fungi isolated from hospital dust and soils of public places at Gulbarga, India

Keratinophilic fungi were isolated from corridor dust of 11 hospitals and soils of 21 public places using a hair baiting technique. A total of 41 species belonging to 24 genera were recovered. Among the dermatophytes and related species, Microsporum gypseum was predominant, followed by Chrysosporium keratinophilum, Trichophyton mentagrophytes, M. nanum and Ch. tropicum. Other species were represented by 32 species belonging to 21 genera. Most of the species isolated are known to be agents of human and animal infection or have been isolated from human and animal lesions. The fungi observed here are discussed in relation to their global distribution.

A survey on the pathogenic fungi in soil samples of potted plants from Sari hospitals, Iran

A total of 23 soil samples of potted plants was collected from hospitals in Sari, Iran. Each sample contained approximately 200 g soil, taken from a depth of 0–10 cm of the pots. Samples were analysed by two different methods. (1) Culture on Sabouraud's dextrose agar medium containing chloramphenicol (SC); cultured fungi were identified by macroscopic and microscopic characterization. (2) The hair-baiting technique (HBT) was used to isolate the keratinophilic fungi. After culture on SC medium, 100% of plates were positive for fungal growth, and in total 1150 colonies with 13 different types of fungi were isolated. Penicillium spp. (52%), Acremonium spp. (20%), Paecilomyces spp. (11.9%), Cladsporium spp. (3.7%) and Aspergillus spp. (3.1%) were the predominant fungal species. Rhizopus spp. (0.1%) were less frequent. Keratinophilic fungi such as Microsporum gypseum, Microsporum cookei and Chrysosporium spp. were isolated using HBT. The presence of pathogenic fungi such as Cladosporium spp, Aspergillus spp, M. gypseum, and M. cookei in potted plants in hospitals represents a potential source of nosocomial infection.