Baiting Technique Research Articles

Incidence of Keratinophilic Fungi from Selected Soils of Vidarbha Region of Maharashtra State, India

One hundred and fifty samples were collected from eleven districts of Vidarbha region of Maharashtra state and screened for the presence of keratinophilic fungi using hair baiting technique for isolation. Seventy-one 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 the ITS1-5.8S-ITS2 rDNA region against the NCBI/Genbank data and compared with deposited sequences for identification purpose. Thirteen species of eight genera were isolated, namely,Auxarthron conjugatum(2.00%),Chrysosporium indicum(14.00%),Chrysosporium evolceanui(2.66%),Chrysosporium tropicum(4.66%),Chrysosporium zonatum(1.33%),Chrysosporiumstate ofCtenomyces serratus(3.33%),Gymnascella dankaliensis(1.33%),Gymnascella hyalinospora(0.66%),Gymnoascoideus petalosporus(0.66%),Microsporum gypseumcomplex (9.33%),Trichophyton mentagrophytes(2.00%),T. terrestre(3.33%), andUncinocarpus queenslandicus(2.00%). This study indicates that the soils of Vidarbha region of Maharashtra may be significant reservoirs of certain keratinophilic fungi.

Isolation of keratinophilic fungi from selected soils of Sanjay Gandhi National Park, Mumbai (India)

One hundred and twenty-five samples were collected from eight different sites in the vicinity of Sanjay Gandhi National Park (SGNP) and screened for the presence of keratinophilic fungi using hair baiting technique for isolation. Seventy-three isolates were recovered and identified. The cultures were identified using macro- and micro-morphological features. Their 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 for identification purpose. Thirteen species of nine genera were isolated viz. Aphanoascus durus (2.4%), Arthroderma corniculatum (1.6%), Auxarthron umbrinum (0.8%), Chrysosporium evolceanui (1.6%), Chrysosporium indicum (16.0%), Chrysosporium tropicum (2.4%), Chrysosporium zonatum (4.0%), Chrysosporium states of Arthroderma tuberculatum (0.8%), Chrysosporium state of Ctenomyces serratus (11.2%), Gymnascella dankaliensis (3.2%), Microsporum gypseum (12.0%), Myriodontium keratinophilum (0.8%) and Trichophyton mentagrophytes (1.6%). Representative of all thirteen species can release the protein in the range of 152.2-322.4 μg/mL in liquid media when grown on human hair in shake flask culture and also decompose 18.4-40.2% of human hair after four weeks of incubation. This study indicates that the soils of SGNP, Mumbai may be significant reservoirs of certain keratinophilic fungi. The keratinolytic activity of these fungi may be playing significant role in superficial infections to man and animals and recycling of keratinic material of this environment.

Epidemiological Study of Keratinophilic Fungi in Baghdad Swimming Pools

The present study was included the isolation of Keratinophilic fungi from water samples taken from 19 indoor public swimming pools in Baghdad during four season (summer ,fall ,spring ,and winter). Isolation period was September 2011and September 2012 . Samples were collected from different places in swimming pool . Sixteen keratinophilic fungal species, belonging to twenty –one genera were isolated using Surface Dilution Plating (SDP) and Hair Bait Technique (HBT) . Aspergillus genus was the most frequent and occurrence (19.84%), followed by Trichophyton (11.60%) and Fusarium (7.59% ) . The most common fungal infection occurred in summer season (42.16%) , and the most frequent month manifestation of species occurred in August (17%) .

Epidemiological Study of Keratinophilic Fungi in Baghdad Swimming Pools

The present study was included the isolation of Keratinophilic fungi from water samples taken from 19 indoor public swimming pools in Baghdad during four season (summer ,fall ,spring ,and winter). Isolation period was September 2011and September 2012 . Samples were collected from different places in swimming pool . Sixteen keratinophilic fungal species, belonging to twenty –one genera were isolated using Surface Dilution Plating (SDP) and Hair Bait Technique (HBT) . Aspergillus genus was the most frequent and occurrence (19.84%), followed by Trichophyton (11.60%) and Fusarium (7.59% ) . The most common fungal infection occurred in summer season (42.16%) , and the most frequent month manifestation of species occurred in August (17%) .

Hyphomycetes in rain water, melting snow and ice

The investigations performed by direct microscopy method or bait technique (fragments of plants, cellophane, snake skin) using rain water flowing from trees, different types of roofs and melting snow and ice allowed determination of 146 species including 55 new to Poland. Aero-aquatic species were predominant and only several belonged to typically aquatic <i>Hyphomycetes</i>.

Keratinolytic and keratinophilic fungi of mangrove's soil and air in the city of Qena and their response to garlic extract and onion oil treatments

Forty-eight species and l variety belonging to 25 genera were collected from 50 soil samples (41 species and l variety and 21 genera) and the atmosphere (27 species and l variety and 14 genera) of mangrove in the city Qena using hair baiting technique at 28"C. Twenty of these species was dermatophytes and closely related fungi. The most common and frequent species of the latter fungi were <i>Aphanoascus fulvescens</i> (telemorph of <i>Chrysosporium keratinophilum), A. terreus (C. indicum). Aphanoascus</i> sp. <i>(C. tropicum)</i> and <i>Chrysosporium xerophilum</i>. Sixty-eight isolates were tested for the abilites for growth on hair - sand medium. Most (73.5 %) had moderate growth rate. All keratinophilie fungi recovered in the present investigation were sensitive to garlic extract and onion oil.

Sludy of the Dermatophytes in the Students Houses of Minia University, Egypt

A survey of dermatophytes and other fungi was carried out in 100 air - dust samples from bedrooms and dinning halls of male and female student resident houses. By hair baiting technique the common dermatophytes were obtained namely <i>Microsporum canis, M. gypseum</i> and <i>Trichophyton mtntagrophytes</i>. Also five species of <i>Chrysasporium</i> were isolated in the following order of dominance <i>C. tropicum, C. keratinophilum, C. indicum, C. pannicola</i> and <i>C. quecnslandicum</i>. By dilution plate method, 37 species representing 20 genera of which <i>Aspergilus niger, A. flavus, Rhizopus nigricans, Penicillium chrysogenum</i> and <i>Cladosporium cladosporioides</i> were most frequently isolated.

English

One hundred and sixty nine Nocardia isolates were recovered from 111 Iraqi soil samples on six cultivation media, using paraffin baiting and dilution techniques. The North of Iraq soil was the richest in Nocardia. Paraffin baiting technique was more efficient in the isolation of Nocardia than dilution technique. Glucose asparagine agar was more suitable for cultivating Nocardia isolates followed by Sabouraud dextrose agar. All the 169 presumptive Nocardia isolates recovered were subjected to primary screening of antibacterial activity, with 10-55 mm inhibition zones against standard Staphylococcus aureus (NCTC 6571) and 10-38 mm inhibition zones against standard Escherichia coli (ATCC 25922). Eleven isolates with the highest antibacterial activities were selected for chemo-taxonomic studies. These isolates exhibited high similar features of 87-89% with Nocardia brasiliensis. Therefore, it is suggested to give proposed names for these new taxa: Nocardia sp.1, N. sp. 2, N. sp. 3, N. sp. 4, N. sp. 5, N. sp. 6, N. sp. 7, N. sp. 8, N. sp. 9, N. sp. 10 and N. sp. 11. . Key words: Secondary metabolite, Nocardia sp., antimicrobial activity.

English

In an endeavor to search for some wild isolates of keratinophilic fungi from the Raipur City of Chhattisgarh, 24 soil samples from different regions of the city were collected and screened using hair baiting technique. A total of six species which belong to two genera were isolated. They are Chrysosporium keratinophilum (27.78%), Chrysosporium pannicola (5.56%), Chrysosporium tropicum (13.89%), Chrysosporium sp. (close to Chrysosporium pannicola) (11.11%), Chrysosporium sp. (16.67%) and Microsporum gypseum (25%). The Chrysosporium species prevalent in this region are able to tolerate temperature of about 40°C and all isolates belong to moderate individual risk group. Key words: Keratinophilic fungi, Raipur, Chrysosporium, Microsporum.

Ant species diversity in ciliary forest and gallery forest areas in central Brazil

Ants make up an important group of insects of the Cerrado in terms of biomass, number of individuals, and ecological impact. The main goals of this study were to quantify the species richness and species diversity of the family Formicidae in two riparian Cerrado physiognomies: Ciliary Forest and Gallery Forest, and to evaluate the effectiveness of two collection techniques for these organisms. The study was conducted during the dry season, comprising the months of May to October 2012, in the municipality of Quirinópolis, Goiás State, Brazil. Samples were made using two capture techniques: sardine baits and pitfall traps. Manual collection was employed to extend the range of faunistic composition of the ants. We collected 24 species of ants in the Gallery Forest and 23 in the Ciliary Forest. The genus Megalomyrmex was the most representative, being present in 49.46% of the samples, and secondly the genus Pheidole, with 15.36% of the total individuals captured. From a total of 10,917 individuals, 9140 were captured using the bait technique, 1738 with traps, and 39 individuals through manual collection. Similar results were found in the Cerrado of Paineiras, with 45 ant species distributed in 16 genera belonging to 5 subfamilies.

Isolation and identification of entomopathogenic nematodes (Nematoda: Rhabditida) from the Eastern Black Sea region and their biocontrol potential against Melolontha melolontha (Coleoptera: Scarabaeidae) larvae

A survey was conducted for entomopathogenic nematodes in various agricultural fields in the Eastern Black Sea region of Turkey. A total of 77 soil samples were collected from 15 distinct geographic areas during 2010-2011. Seven entomopathogenic nematode isolates (ZET02, ZET04, ZET09, ZET28, ZET31, ZET35, and ZET76) were detected from the soil samples (9.1% positive) using the Galleria baiting technique. Morphological and molecular characterizations of the isolates were performed for species identification. Five isolates were identified as Heterorhabditis bacteriophora (ZET02, ZET04, ZET09, ZET28, and ZET35) and 2 isolates were identified as Steinernema feltiae (ZET31 and ZET76). The efficacy of all isolates was tested on Melolontha melolontha larvae in plastic boxes and pot experiments. Different concentrations of nematodes at 0, 500, 1000, or 2000 infective juveniles (IJs)/mL and 2 different temperature regimes (15 and 25 °C) were used. One hundred percent mortality was obtained from the ZET09 and ZET35 isolates at a concentration of 2000 IJs/mL at 25 °C. The same isolates also provided 100% protection with 100 IJs/cm^2 in strawberry planted pot experiments.

Ultrastructure of symbiotic germination of the orchid Dendrobium officinale with its mycobiont, Sebacina sp.

Dendrobium officinale is an endangered epiphytic orchidaceous medicinal plant. Similar to other orchid plants, the seed germination of D. officinale under natural conditions depends nutritionally upon mycorrhizal fungi. The compatible fungi have been isolated from D. officinale protocorms using in situ seed baiting technique in our previous studies. However, the interaction between seed germination of D. officinale and its mycobiont is still unclear. In the present study, we investigated the morphological changes of seed and fungus during the symbiotic germination using a light microscope and transmission electron microscope. Seeds of D. officinale have no conspicuous suspensor cells. The fungus enters into the embryo cell through the posterior end of the embryo and colonises the cortical cell in the first stage of germination (Stage 1). Then, the hyphae form pelotons with the protocorm development (Stages 1–3). After protocorm formation, the reinvaded fungal hyphae conspicuously decrease. Invaded hyphae lose bioactivity, form clumps and start degeneration at Stage 4 or 5 (seedling development). When penetrating the neighbouring cortical cell, the fungal hyphae constrict to collar shape at the contacted site and follow by swelling in the apex. Our study suggested that fungi trigger protocorm development and concomitant reserve utilisation during the symbiotic germination.

Comparative analysis of keratinophilic fungi from the soils of Khardung and Khardung La (Ladakh), India

The present research work was undertaken to study the diversity of keratinophilic fungi from the unexplored soils of Khardung and Khardung La (Ladakh). This was done with a view to evaluate the relationship between the keratinophilic fungi and the keratinous material present in the soil. The soil samples examined from Khardung La were found to be 80% positive for keratinophilic fungi, whereas that of Khardung were 100% positive. A total of 29 keratinophilic species belonging to 13 genera were recovered by keratin bait technique from the soils of Khardung. However, from Khardung La, only 7 species belonging to 3 genera were recovered. The recovered mycokeratinophiles consisted of 5 species each of Aspergillus and Fusarium, 4 species of Chrysosporium, 3 species of Paecilomyces, 2 species each of Acremonium, Gliocladium and Penicillium and 1 species each of Beauveria, Cladosporium, Embellisia, Emericella, Epicoccum, and Sagenomella. Comparison of diversity indices showed a significant difference in species richness between the two sites i.e., Khardung (29) and Khardung La (7). The values of Shannon‟s diversity index and Simpson‟s diversity index were 3.27 and 0.96 respectively for Khardung, whereas these values were 1.88 and 0.84 respectively for Khardung La. Sorenson‟s similarity index showed a value of 38.89% for these two sites, which indicates that some of the species present in Khardung La soils are also prevalent in Khardung soils.

Evaluation of various techniques for the detection of Nocardia species in clinically evident cases of pulmonary and extrapulmonary tuberculosis

To determine the risk factors, microbiological features, clinical features and other epidemiological characteristics of Nocardia asteroides, we evaluated 500 patients with suspected pulmonary and extra pulmonary tuberculosis seen over a period of 18 months. Samples were subjected to microscopy and culture using standard protocols. Out of 500 samples, 5 (1%) were found to be positive for Nocardia asteroids on the basis of growth characteristics and biochemical reactions. All the 5 (100%) cases were detected by McClung's Carbon free broth by paraffin baiting technique. Paraffin baiting technique had significantly higher efficacy (p<0.0001) as compared to Nutrient agar, Sabouraud's dextrose agar, Lowenstein-Jensen medium, paraffin agar and Gelatin agar. Antimicrobial susceptibility testing was done by disk diffusion method in blood agar and Mueller-Hinton agar plates, as recommended by the National Committee for Clinical Laboratory Standards. Amikacin and imipenem showed the 100% sensitivity. A diagnosis of nocardiosis may have direct benefits to the patient and physician by providing a clinical diagnosis and limiting extensive diagnostic evaluations. It may also reduce the use of empirical therapy, which could be ineffective and potentially harmful.

Biodegradation of Keratin from Chicken Feathers by Fungal Species as a Means of Sustainable Development

Keratinolytic microorganisms have a great importance in feather waste degradation and its use for improvement of livestock feed and production of protein hydrolysates. Annually several thousand tons of feather wastes are discharged into the surrounding environment as a by-product of commercial poultry processing. Microorganisms could minimize regulatory problems of uncontrolled accumulation of waste feathers. This residue is almost pure keratin, which is not easily degradable by common proteolytic enzymes. The present study deals with identification of fungi that play a significant role in the degradation of chicken feather and keratin degradation ability of the isolated fungi. Feathers of broiler chicken were collected from Jaggi poultry farm, Mandir Hasaud, Raipur. Fungi were isolated by feather baiting technique. Feathers were inoculated in Sobouraud Dextrose Agar (SDA) medium and their pure culture was prepared. Fungus were identified by Lacto phenol cotton blue staining method as Trichoderma, Gliocladium, Fusarium, Syncephalastrum, Mucor, Aspergillus Flavus. The pure culture were grown in mineral media with 500 mg of feathers as a sole source of Nitrogen and Carbon and incubated for the period of 25 days. At 5 days intervals, the biochemical changes associated with biodegradation was evaluated by analyzing the culture filtrate. The release of Nitrate, Cystine, Cysteine and methionine components during the process of biodegradation was studied which proved the efficient degradation of keratin. There was also a change in pH of the medium towards alkalinity. Mucor and Aspergillus Flavus were the most powerful bio remedial fungus in the current study. With an increasing world-wide concern for the environment it is possible to use these six fungus for the degradation of enormous quantity of waste feathers. Biodegradation leads to recycle the wastes and thus maintaining the environmental quantity of the biosphere.

A STUDY ON ROLE OF KERATINOPHILIC FUNGI IN NATURE: A REVIEW

The chapter reviews available data on the ability of keratinophilic fungi and include a variety of filamentous fungi mainly comprising Hyphomycetes and several other taxonomic groups. Hyphomycetes include dermatophytes and a great variety of dermatophytes filamentous fungi. Most of the latter occur as saprophytes in soil,and some are plant pathogens. Chrysosporium species are the commonest dermatophytes filamentous fungi and are predominantly recovered from soil and other natural substrata by hair baiting technique C. tropicum and C. panniocola have been frequently isolated from human and animal’s skin lesions but their etiological relationship has not been established. The criteria considered important for evaluating the role of dermatophytes filamentous fungi in skin infection are the demonstration of mycelial elements in direct microscopy of skin scrapings or biopsy compatible with those in culture repeated positive cultures from clinical material. The Oils plant has been reported to have antibacterial, antifungal, antiviral, antipurasitic and antidermatophytic properties. It is now considered as a valuable source of natural products for development of medicines against various diseases and also for development of Industrial products. In this review is a compilation of updated information on plant essential oils with antifungal properties.

Microsatellite markers to monitor a commercialized isolate of the entomopathogenic fungus Beauveria bassiana in different environments: Technical validation and first applications

Here, we report on the application of five previously developed microsatellite markers (simple sequence repeats, SSRs) to monitor an isolate of the entomopathogenic fungus Beauveria bassiana (Bals.) Vuill. in different environments. Discriminatory power of these SSR markers was assessed in two commercialized B. bassiana isolates as well as in 16 B. bassiana isolates from a world-wide collection, and three of the five SSR markers were estimated to allow a confident discrimination among the given isolates. Sensitivity thresholds of 0.1pg DNA were subsequently determined for all SSR markers in case pure genomic fungal B. bassiana DNA was used as a template for PCR assays, but threshold levels varied depending on the environment (soil, plant) of the PCR assay. Furthermore, presence of a commercialized B. bassiana isolate was monitored via these SSR markers in three different types of potting substrates over a period of 14weeks. With two SSR markers, strain-specific products were detected up to 14weeks after application of B. bassiana to the substrate. Infectivity of B. bassiana conidia in the respective soil samples was confirmed by the Galleria baiting technique. Together these results indicate that molecular markers like SSRs specific for commercialized strains of entomopathogenic fungi are important tools to monitor a particular fungal strain in complex environmental samples such as bulk soil or plant DNA.

Isolation of Keratinophilic Fungi and Aerobic Actinomycetes From Park Soils in Gorgan, North of Iran

Background: Keratinophilic fungi are a group of fungi that colonize in various keratinous substrates and degrade them to the components with low molecular weight. This study was conducted to determine the prevalence of keratinophilic fungi and aerobic Actinomycetes in soil of city parks in Gorgan. Objectives: In this study, we surveyed the city park soils of Gorgan (a northern province of Iran) to determine the identities and diversity of soil aerobic Actinomycetes, keratinophilic and non-keratinophilic fungi. Materials and Methods: A total of 244 soil samples were collected from 22 different parks of Gorgan, North of Iran. The samples were collected from the superficial layer with depth not exceeding than 0-10 cm in sterile polyethylene bags. We used hair bait technique for isolation keratinophilic fungi. The colonies identified by macroscopic and microscopic characterization after slide culturing. Actinomycetes were isolated by antibiotic dilution methods and detected by using physiological tests such as Lysozyme, Casein, Xanthine, Hypoxanthine, Gelatin, Urea Broth, and modified acid-fast stain. Results: Totally, 75 isolates of aerobic Actinomycetes were detected that Actinomaduramadurae and Nocardiaasteroides were the most prevalent strains, with 14.66 and 28% prevalence respectively. Microsporumgypseum was more frequent than other keratinophilic fungi (22.96%) and Aspergillus spp. was the most species of saprophyte fungi (15.92%). Conclusions: This study showed that the collected soil from studied areas was rich of keratinophilic fungi and Actinomycetes, therefore hygiene protocol should be taken to prevent the spread of pathogenic and saprophytes fungi in the environment of susceptible person.

Pollen baiting facilitates the isolation of marine thraustochytrids with potential in omega-3 and biodiesel production

Marine heterotrophic microbes are capable of accumulating large amounts of lipids, omega-3 fatty acids, carotenoids, and have potential for biodiesel production. Pollen baiting using Pinus radiata pollen grain along with direct plating techniques were used in this study as techniques for the isolation of oil-producing marine thraustochytrid species from Queenscliff, Victoria, Australia. Thirteen isolates were obtained using either direct plating or using pine pollen, with pine pollen acting as a specific substrate for the surface attachment of thraustochytrids. The isolates obtained from the pollen baiting technique showed a wide range of docosahexaenoic acid (DHA) accumulation, from 11 to 41% of total fatty acid content (TFA). Direct plating isolates showed a moderate range of DHA accumulation, from 19 to 25% of TFA. Seven isolates were identified on the basis of 18S rRNA sequencing technique as Thraustochytrium species, Schizochytrium species, and Ulkenia species. Although both methods appear to result in the isolation of similar strains, pollen baiting proved to be a simpler method for the isolation of these relatively slow-growing organisms.

Microsporum fulvum IBRL SD3: As Novel Isolate for Chicken Feathers Degradation

Keratinous wastes have increasingly become a problem and accumulate in the environment mainly in the form of feathers, generated mainly from a large number of poultry industries. As keratins are very difficult to degrade by general proteases, they pose a major environmental problem. Therefore, microorganisms which would effectively degrade keratins are needed for recycling such wastes. A geophilic dermatophyte, Microsporum fulvum IBRL SD3 which was isolated from a soil sample collected from a chicken feather dumping site using a baiting technique, was capable to produce keratinase significantly. The crude keratinase was able to degrade whole chicken feathers effectively. The end product of the degradation was protein that contained essential amino acids and may have potential application in animal feed production. Thus, M. fulvum could be a novel organism to produce keratinase for chicken feathers degradation.