ISSN: 2456–4397 RNI No.  UPBIL/2016/68067 VOL.- VII , ISSUE- II May  - 2022
Anthology The Research
Aero-Mycoflora of Ginneries in Lucknow
Paper Id :  16004   Submission Date :  2022-05-20   Acceptance Date :  2022-05-22   Publication Date :  2022-05-25
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Virendra Kumar Tiwari
Principal
Botany
Pioneer Mahila Mahavidyalaya
Barabanki,Uttar Pradesh, India
Abstract
In a recent review on the fungal diversity of indoor environments of occupational importance in India, it has very clearly been stressed by Misra et al. (2002) that there is a need for undertaking extensive and intensive studies to know the indoor fungal flora of places of occupational importance and to study the implications of the fungi occurring in such environments both on material and on life indoors. In order to generate information to fill in similar gaps in such knowledge, a survey of Ginneries of Lucknow, both temporary and permanent situated in
Keywords Ginnery, Air borne fungi, Occupational Environment, Aerobiology, Aero-Mycoflora.
Introduction
Air borne fungal spores play an important role in plant disease forecasting and causing respiratory allergic disorders. This paper deals with the study of aero-mycoflora of ginneries in Lucknow and describes the identification of the fungi isolated and spores recovered from such environments
Objective of study
Allergenic effects of fungi are much not known & that too of those that inhabit/or living in the air of Ginneries.The studies wee undertaken to find out the mycoflora of of ginneries in Lucknow using different methods/Media.Few selected fungi were also tested for their allergenic behaviour on human beings.
Review of Literature
Fungi belonging to various groups were isolated. Almost, there was no difference in the number of fungi isolated from the outdoors and indoors of the ginneries of Lucknow. Ginneries, both temporary and permanent, yielded 35 fungal forms including sterile mycelia whereas from outdoors 33 fungi were isolated (Table-1). Ten fungi were isolated using cellulose as the sole carbon source. Of these, four were the species of Aspergilli--Aspergillus flavus, A. fumigatus, A. niger and A. ochraceus. And, one Ascomycetous genus Chaetomium was isolated. Others were the species of Cladosporium, Fusarium, Rhizoctonia, Rhizopus and Stachybotrys (Table-2). The fungal spores recovered using Rotorod and Gravity slides were 17 in number including various hyaline and dark coloured mycelia. This data is presented in table- 3. The spores of Aspergillus (75%), Fusarium (75%), and Hyphal fragments (75%) were the most commonly encountered types followed by the spores of Penicillium (68%), Curvularia (60%), Epicoccum (60%), Pithomyces (60%), and Smut spores (60%). The frequency of occurrence of these spores is shown in parentheses against each. Other spores were observed in various frequencies ranging from 15-45 per cent. These are briefly described below.
Methodology
For determining the fungi in the indoor air of Ginneries, following sampling techniques were adopted. THE GRAVITY SLIDE METHOD Slides were used for capturing and investigating the spores of indoor air. The slides coated with glycerin jelly were exposed regularly at various places and the settled spores were examined and quantified. THE GRAVITY PETRI-DISH METHOD Sterile Petri-dishes of 10 cm diameter containing various synthetic and natural media such as Czapek-dox agar (Sucrose-30 gm, MgSO4.7H2O - 0.5 gm, KCl – 0.5 gm, NaNO3 – 2.0 gm, K2PHO4 – 1.0 gm, FeSO4 – 0.01 gm, Agar-Agar – 20 gm, distilled water – 1000 ml, pH – 7); Sabouraud agar (Peptone – 10 gm, glucose – 40 gm, KH2PO4 – 1.75 gm, MgSO4 – 0.75 gm, Agar-Agar – 20 gm, distilled water – 1000 ml, pH-8); Potato dextrose agar (Potato-slices – 200 gm, dextrose – 20 gm, Agar-Agar – 20 gm, distilled water – 1000 ml); and Malt-extract agar (Malt-extract – 25 gm, Agar-Agar – 20 gm, distilled water – 1000 ml, pH-7) containing antibacterial agent (Streptomycin sulphate, 0.5 gm/L) were exposed in the indoor environments of different Ginneries for varying period of time ranging from 2-5 min. Exposed Petri-dishes were incubated at 28 ±1ºc for 3-7 days. The colonies developed were counted and isolations were made on the slants containing appropriate media for identification. In order to observe the substrate relationship, carbon source of Czapek-dox agar was replaced by cellulose. Petri-plates were similarly exposed and incubated to isolate and observe the fungi growing on them. Rotorod air sampler (Manufactured by T. Mohan Rao, Pune) was also used for catching the fungal spores of the indoor air. The sampler has collecting arms made up of brass having 0.159 cm cross sectional area. It is square in shape and slightly bent inwards. The vertical arms are 6 cm long and 4 cm apart from rotating axis. This was run by dry battery for ½-1 hr at various Ginneries. The cello-tape was mounted on the collecting arms of the sampler with jelly applied on them as adhesive for spores. After definite exposure, the tapes were mounted on the slide and immediately melted jelly was poured over the slide and covered with the cover slip. After drying, the slides were scanned under light microscope for identification. Fungi so obtained were identified with the help of standard keys and monographs (Barron, 1968; Tandon, 1968; Ellis, 1971, 1976; Raper and Fennell, 1977; Sutton, 1980; Raper and Thom, 1984).
Sampling

Fungi belonging to various groups were isolated. Almost, there was no difference in the number of fungi isolated from the outdoors and indoors of the ginneries of Lucknow. Ginneries, both temporary and permanent, yielded 35 fungal forms including sterile mycelia whereas from outdoors 33 fungi were isolated (Table-1).
Ten fungi were isolated using cellulose as the sole carbon source. Of these, four were the species of Aspergilli--Aspergillus flavus, A. fumigatus, A. niger and A. ochraceus.  And, one Ascomycetous genus Chaetomium was isolated. Others were the species of Cladosporium, Fusarium, Rhizoctonia, Rhizopus and Stachybotrys (Table-2).
The fungal spores recovered using Rotorod and Gravity slides were 17 in number including various hyaline and dark coloured mycelia. This data is presented in table- 3.
The spores of Aspergillus (75%), Fusarium (75%), and Hyphal fragments (75%) were the most commonly encountered types followed by the spores of Penicillium (68%), Curvularia (60%), Epicoccum (60%),  Pithomyces (60%), and Smut spores (60%). The frequency of occurrence of these spores is shown in parentheses against each. Other spores were observed in various frequencies ranging from 15-45 per cent.  These are briefly described below.
Description of Spores / Mycelium Collected on Slides and or Rotorod Air Sampler
Alternaria
Conidia brown to golden brown, smooth with transverse and longitudinal septa, measuring 15-60 x 8-10 µm were seen. Its frequency of occurrence was 45%.
Aspergillus
Smooth to echinulate spores of various colour and sizes with distorted conidial heads were frequently seen (75%). 
Bispora
Conidia dark, bi-celled with thick black septum, measuring 7 x 7 µm in size. It occurred in moderate frequency (45%).
Cercospora
Conidia golden yellow, elongated, several celled, measuring 50-62 x 6-9 µm. Its frequency of occurrence was low (23%).
Chaetomium
Globose to sub-globose, limoniform ascospores measuring 9-11 x 8-10 x 6-8 µm were seen in very low frequency-15%. 
Cladosporium
Conidia variable in size and shape, ovoid to cylindrical, light yellow to dark brown with distinct scar of attachment on either of the body, measuring 10-30 x 6-10 µm. Its frequency of occurrence was 60%.
Curvularia
Conidia typically curved, 3-5 celled, dark yellow to brown, end cell lighter, central cell enlarged, measuring 5-13 µm. It occurred moderately (60%).
Drechslera
Conidia with pseudosepta, pale to dark brown, variable in shape and size measuring 22-34 x 15-23 µm. Its frequency of occurrence was moderately low (45%).
Epicoccum
Conidia globose, dark brown, several celled, measuring 14-22 µm in diameter. Its frequency of occurrence was 60%.
Fusarium
Sickle shaped macroconidia of variable size and number of septa were seen, measuring 35-60 x 5-8 µm.  Their frequency of occurrence was 75%.
Hyphal Fragment
Septate hyphal fragments, both hyaline and pigmented of varying dimensions were often encountered. Their frequency of occurrence was moderately high (75%).
Nigrospora
Dorsiventrally compressed, globose, unicelled, conidia measuring 12-15 µm were seen at the frequency of 45%.
Penicillium
Globose, smooth to echinulate spores on phialides were seen, which were thought to be of Penicillia or like fungus at the frequency of 68%.
Pithomyces
Echinulate dark coloured spores measuring 12-20 x 4-8 µm were seen in 60% frequency.
Rust Spores
Occurred in groups, yellow to brown in colour, ovoid to spherical, mostly bicelled teliospores were seen at the frequency of 45%.
Smut Spores
Spores variable in size, dark brown, rounded, echinulate, with thick wall, measuring 4-5 µm in diameter were at the frequency of 60%.
Trichothecium
Conidia bi-celled, one cell larger than the other, measuring 10-17 x 8-10 µm. Its frequency of occurrence was 45%.

Tools Used Rotorod air sampler (Manufactured by T. Mohan Rao, Pune) was also used for catching the fungal spores of the indoor air. The sampler has collecting arms made up of brass having 0.159 cm cross sectional area. The vertical arms are 6 cm long and 4 cm apart from rotating axis. This was run by dry battery for ½-1 hr at various Ginneries. The cello-tape was mounted on the collecting arms of the sampler with jelly applied on them as adhesive for spores. After definite exposure, the tapes were mounted on the slide and immediately melted jelly was poured over the slide and covered with the cover slip. After drying, the slides were scanned under light microscope for identification.
Fungi so obtained were identified with the help of standard keys and monographs (Barron, 1968; Tandon, 1968; Ellis, 1971, 1976; Raper and Fennell, 1977; Sutton, 1980; Raper and Thom, 1984).
Result and Discussion

Out of 35 fungal forms, 13 were the Aspergilli, both from the outdoor and indoor environments of ginneries in Lucknow. The other Deuteromycota members isolated were--Alternaria alternata, two species of CladosporiumCurvularia lunata var. aeriaDrechslera hawaiiensis, two species of Fusarium, and one species each of Monilia, Nigrospora, Paecilomyces, Penicillium, Periconia, Pithomyces, Rhizoctonia, Stachybotrys, Trichoderma and Trichothecium.  Only one member of Ascomycota--Chaetomium globosum was isolated both from the indoor and outdoor environments.  This fungus was also isolated on the cellulose agar medium. One Coelomycetes--Pestalotia pezizoides was also found both from the indoor and outdoor environments of Ginneries.
Only two genera of Zygomycota, Mucor and Rhizopus with one species each were isolated (Table-1).
Cladosporium herbarum and Rhizoctonia solani were not found from the air at the outdoors of Ginneries during the present study.

Conclusion
The occurrence of 35 and 33 species of fungi from inside of the ginneries and their outside environment in Lucknow indicates that there is a diversity of fungi both at the in-doors and out-doors of ginneries. The present study also indicated that the dust produced during the process of ginning, both at the permanent and temporarily installed cotton gins (machines) as well as the hand-operated stringed tool or a cotton gin that a ginner uses produce a lot of dust and spores that often at times it becomes difficult to go inside the ginneries. The environment becomes dark and suffocating. On Petri-plates or slides exposed during the time of ginning or little after the ginning is over, the fungal colonies appear in such a high number that it almost become difficult to count and isolate them. This indicates the enormousness of the fungal spores in the dust resulting due to ginning. Hence the little variation in the number of forms recovered from the indoors and outdoors in the present study is not surprising. The current observation is in conformity with the results of other workers of other and a similar environment (Flour Mills) of Lucknow (Gupta and Singh, 1983; Shukla and Misra, 1984, 1985; Shukla, 1987; Misra et al., 1989; Misra and Shukla, 1989; Misra and Jafar, 1991). Other workers, both from India and abroad, like Mukerji et al. (1969), Rati et al. (1980), Tilak et al. (1980, 1981), Gaur and Kasana, (1981), Singh et al. (1981), Pande (1981), Jayaprakash and Ramalingam (1981, 1983), Gaur and Kala (1984), Rogers (1984), Sinha et al. (1984), Tilak (1984), Tilak and Saibaba (1984) and Misra et al. (2002) also had similar conclusions of their study as of the present one. The dominance of Aspergilli in ginneries is in agreement with the results of Jayaprakash et al. (1978), Tilak and Chakre (1979), Jayaprakash and Ramalingam (1981, 1983), Santra and Chanda (1981), Singh (1981) and Mehta and Sandhu (1983). Aspergillus flavus, A. fumigatus and A. niger were the most frequent and dominant species of Aspergilli in the ginneries. Sinha et al. (1984) had also reported A. niger as a common fungal form in the air of Calcutta. The isolation of Chaetomium globosum on cellulose medium indicates its affinity to the medium. Hence, its occurrence in the ginneries where cellulose materials are abundant is logical. Not only this, but Chaetomia are also known to have ability to degrade cellulose materials (Alexopoulos et al., 1996). Stachybotrys atra was recovered from those Ginneries which often gin old, used and poor quality cotton brought by the people who are economically poor and who do not keep the cotton filled articles safe and clean to avoid fungal infestation. Therefore, such cotton is bound to increase the fungal spores/ dust, etc. in the Ginneries and consequently the fungi over the isolation medium, as has been presently observed. Most of the fungal spores recorded during this study had also been seen by Shukla (1987) and Hashim (2002) from the Cobbler shops and Flour Mills in Lucknow environment, respectively. However, the frequency of occurrence of different spores was not the same. Rust spores, as captured from Ginneries had also been reported by Tilak et al. (1981), and Tilak and Saibaba (1984). The observation of spores like those of Aspergillus, and Fusarium in higher frequencies followed by that of Cladosporium, Curvularia, Epicoccum, Pithomyces and Smut spores (Table-3) indicate that plant materials and accompanying dust are the source of these fungal spores (Garrett, 1951, 1956; Madeline and Linton, 1974). Similar were the observations of workers like Verma et al. (1981), Verma and Kamal (1982), and Shukla (1987). Out of 10 fungi recovered on the cellulose medium indicated their close relationship with the cellulosic substrate. Aspergilli (4 species) were the dominant ones. Aspergilli are known to degrade cellulose and hence their dominance even on cellulose medium could be justified. Chaetomium, Fusarium, Rhizoctonia and Rhizopus are also reported to be good cellulose decomposer (White et al., 1948; Alexopoulos et al., 1996). The observations recorded during this study also indicated that, in order to have a complete picture of genera and species in a given environment, all possible techniques and media should be used.
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