Rituparna Saha1, Susmita Sett1, Tribeni Chatterjee1, Gouri Basak1, Anirban Roy2, Krishnendu Acharya1*
1Molecular and Applied Mycology and Plant Pathology Laboratory, Department of Botany,
University of Calcutta, Kolkata, West Bengal – 700019, India.
2West Bengal Biodiversity Board, Prani Sampad Bhawan, 5th Floor, LB - 2, Sector - III, Salt Lake City, Kolkata, West Bengal 700106, India.
Among the macrofungal specimens collected from various ecological regions of West Bengal, six species under four genera (Auricularia, Turbinellus, Craterellus and Clavaria) have been identified through macro and micro morphological investigation. Out of these, three species (Auricularia cornea Ehrenb., Craterellus shoreae Hembrom, K. Das, A. Parihar and Buyck and Clavaria zollingeri Lev.) are reported for the first time from West Bengal. All the six species are detailed with amplified description having illustrations, photographs and necessary taxonomic notes for easy identification.
West Bengal of Eastern India, is unique for its varied topography and climate that results the ecological regions like Himalayan hills, Terai-Dooars, gangetic plains, lateritic region and coastal plains. These zones with different ecological conditions have various forest types which are very much conducive for diversified microhabitats of macromycetes. Since the last eighteen years, our laboratory has been engaged in exploration of macrofungi in different parts of West Bengal of which a good many species have been contributed in the macromycetes of West Bengal 1‒7. In continuation to our earlier contribution, the present work deals the detailed account of six macrofungal species [Auricularia fuscosuccinea (Mont.) Henn., Auricularia cornea Ehrenb. and Auricularia nigricans (Sw.) Birkebak Looney and Sánchez- Garcia, Turbinellus floccosus (Schwein.) Earle ex Giachini and Castellano, Craterellus shoreae Hembrom, K. Das, A. Parihar and Buyck and Clavaria zollingeri Lev.].
MATERIAL AND METHODS:
The macro-fungal specimens were collected during monsoon to post monsoon season from June to October from different forest and road side areas of West Bengal, India. Necessary field notes of the collected specimens like place and date, habit, habitat, shape, size and colour of the fruit body, types of fruit body with the attachment to substratum, presence/absence of hairs, types of hymenium region, presence/absence of stipe and stipe attachments were taken along with colour photographs for future references. The fruit bodies were carefully separated with the help of scalpel and chisel from the substratum. Then each collection was wrapped with tissue papers and isolated in a box to avoid mixing of spores. The collected specimens were dried in a hot air drier to remove moisture. Microscopic characters were noted by making transverse sections of these dried materials by mounting and staining in 10% KOH, Congo red and Melzer’s reagent and observing it under microscope. Amyloidity/ Non-Amyloidity/Dextrinoidity of the microscopic features were observed using Melzer’s reagent. Microscopic characters like pileipellis cells, stipitipellis cells, hyphal system, presence or absence of clamp connections, presence or absence of secondary septa, presence or absence of medulla, size of hairs, basidia, basidiospores, cystidial element, presence or absence of gloeopherous hyphae, presence or absence of guttulae in the spores etc. were noticed under Carl Zeiss AX10 Imager A1 phase contrast microscope for identifications of the specimens following standard keys and published literatures8‒13. Methuen Handbook of colour was used for colour terms and codes of specimens14. To calculate the dimensions of basidiospores thirty measurements were taken from each specimen. The Q value is denoted by Length/breadth ratio. The measurement of mean Q value (Qm) was done by dividing total sum of Q value by total number of spores observed. Outline of all identifying characters were drawn using camera lucida and 0.1 mm rotring pen was used to trace the lines. To preserve the specimens standard protocol was followed15. The voucher specimens were deposited systematically at Calcutta University Herbarium (CUH), Kolkata, India.
RESULTS AND DISCUSSIONS:
Auricularia fuscosuccinea (Mont.) Henn.
Basidiomata Solitary to gregarious :
mostly pileate, substipitate to sessile, gelatinous when fresh, thin membranous when dry, orbicular, 32‒36 mm wide and 2–3 mm thick; abhymenial surface reddish orange (7B6), slightly folded, margin entire; hymenium brownish orange (6C3), smooth, lubricous.
Zona pilosa 7.74–25.8 µm wide; hairs scattered 21.5–34.4 × 3.44–5.17 µm, vinaceous brown-hyaline, thick walled, gregarious, tips acute. Zona compacta 15–47.3 µm wide, densely packed, individual hyphae indistinguishable. Zona subcompacta superior 17.2–21.5 µm wide. Zona laxa superior 51.6–193.5 µm wide, hyphae 3–4 µm wide. Medulla 43–94.6 µm wide, often pigmented, composed of a single band of hyphae, hyphae 2–3 µm wide, oriented parallel to the surface, slightly abhymenial in position. Zona laxa inferior 150–245 µm wide, hyphae 3–4 µm in diameter. Zona subcompacta inferior 43–68.8 µm wide. Hymenium 73.1–86 µm thick, pigmented, yellowish brown, clamp connection present. Basidia 30.17–56.03 × 4.3–5.1 µm, cylindrical, transversely 3 septate, epibasidium with oil guttles, sterigma prominent, 4.3–9.03 µm × 0.43–0.86 µm. Basidiospores 8.6–9.46–11.38–12.4(–13.76) × 3.87–4.9–5.16(–6.02) µm, cylindric˗allantoid, smooth walled, curved at tip, 1–2 oil droplets, Q = 1.6–3.33, Qm = 2.26.
Habit and habitat:
Solitary to gregarious, grown on dead wooden log of angiosperms.
INDIA, West Bengal, Kolkata, Joka, 22.453˚N 88.301˚E, alt. 8m, A.K. Dutta, 13 May-2012, CUH AM303; INDIA, West Bengal, Targhera, Jalpaiguri, 24.382˚N 87.846˚E, alt. 137m, K. Acharya, A. Roy and R. Saha, 19 September-2017, CUH AM306.
Auricularia fuscosuccinea (Mont.) Henn. is well characterized by its orbicular, reddish orange (7B6), thin membranous basidiocarp; smooth hymenial surface; presence of scattered hairs measuring 21.5–34.4 × 3.44–5.17 µm; distinct medullary layer and basidiospores measuring 8.6–13.76 × 3.8–6.02 µm with a Qm value of 2.26. This present taxon was previously reported from Cuba16, Columbia11 and Brazil17. Our collection is nicely matched with the specimen reported from Columbia. This specimen mostly matches with the description reported from Cuba, except for having slightly larger basidia (45–59 × 3.0–6.5 µm vs 30.17–56.03 × 4.3–5.1 µm). The collection reported from Brazil mostly matches with our collection, however differs on the basis of thickness of zona pilosa layer (40–100 µm vs 7.74–25.8 µm), which may be due to difference in climatic and habitat conditons. Among macro˗microscopically related taxa Auricularia cornea Ehrenb. differs by having coriaceous basidiocarp with longer hairs (up to 380 µm) and larger basidiospores (13.5–17.5 × 6–7.5 µm) 16 and A. nigricans (Sw.) Birkebak Looney and Sánchez- Garcia differs by having thick zona pilosa layer (600–700 µm) 17.
Auricularia cornea Ehrenb.
Basidiomata solitary or caespitose, sessile to substipitate, firm gelatinous when fresh and rough coriaceous when dry, pileate; 5‒17 mm wide and 1.5–2 mm thick; abhymenial surface pilose, reddish brown(9D4) to greyish red (10D4), margin entire; hymenial surface smooth, greyish violet (19D4) to greyish magenta (14D4).
Zona pilosa 181.83–314.07 µm wide, hairs 60.34–349.11 × 4.31–8.18 µm, brown, hyaline, thick walled, wall thickness 2.5–3 µm, dense often aggregates in short tufts, apex acute. Zona compacta 30.17–86.2 µm wide, brownish, densely packed, individual hyphae indishtinguishable. Zona subcompacta superior 17.2–64.5 µm wide. Zona laxa superior 129.3–258.6 µm wide, hyphae 2–3 µm wide. Medulla 21.55–86.2 µm wide, composed of single band of hyphae, hyphae 1.5–2 µm wide. Zona laxa inferior 64.45–172.4 µm wide, hyphae 2–3 µm wide. Zona subcompacta inferior 25.86–86.2 µm wide. Hymenium 47.41–64.65 µm thick, pigmented, brown, clamp connection present. Basidia 30.14–51.72 × 4.31–5.17 µm, cylindrical, transversely 3 septate, epibasidium with prominent sterigmata, sterigma 8.62–21.55 × 2.1 µm wide. Basidiospores 10.7–12.93–13.93–15.08–(16.80)17.24 × 4.31–5.77–(6.03)7.75µm, cylindric, smooth walled with curved tip, 1–2 oil droplets, Q = 1.6–3, Qm = 2.46.
Habit and habitat: Solitary to gregarious, grows on dead wooden log of angiosperms.
Specimen examined: INDIA, West Bengal, East Medinipur, Darua, 21.88065˚N 87.3344˚E, alt. 6m, K. Acharya, 1 November-2018, CUH AM304.
Auricularia cornea Ehrenb. is distinctly characterized by its pileate firm reddish brown (9D4) basidiocarp with smooth hymenial surface; brown hyaline hairs ranging from 60.34–349.11 × 4.31–8.18µm; presence of thin medullary layer; and basidiospores measuring 10.7–17.4 × 4.31–7.75µm with a Qm value 2.4. Auricularia cornea has been reported for first time from West Bengal. There are earlier reports on the occurrence of present taxon in Thailand18, Mariana Island16, Hawaii island10, Russia19 etc. Our collection mostly matches with the description reported from Russia except having slightly longer hairs (150–500 × 4.3–7.8µm vs 60–349 × 4–8µm). The descriptions reported from Hawaii, Mariana and Thailand differs by having longer basidia (100–110 × 4–5 µm, 52–69 × 3–6 µm, 55–66 × 3–4 µm respectively vs 30–51 × 4–5µm) and thick medullary layer (up to 130 µm vs 21–86µm) which may be due to differences in climatic and geographical conditions. Among macro˗microscopically related taxa, Auricularia fuscosuccinea (Mont.) Henn. differs by having thin transparent basidiocarp, thin pillose zone (20–90 µm) and smaller basidiospores (8–10 × 4.2–6.5 µm)11; A. nigricans (Sw.) Birkebak Looney and Sánchez- Garcia differs by having thick zona pilosa layer (600–700 µm) and thick medulla layer (40–230 µm)17.
Auricularia nigricans (Sw.) Birkebak Looney and Sánchez- Garcia Figs. 1(VII‒IX)
Fig.1: (I‒II) Fresh basidiomata of Auricularia fuscosuccinea, scale bars: 10 mm, (III) Microscopic characters of Auricularia fuscosuccinea (a) Cross-section of basidioma, (b) Hairs, (c) Basidia, (d) Basidiospores, (e) Generative hyphae, scale bars: a=100 μm, b–e = 5 μm; (IV‒V) Fresh basidiomata of Auricularia cornea, scale bars: 10 mm, (VI) Microscopic characters of Auricularia cornea (a) Cross-section of basidioma, (b) Hairs, (c) Basidia, (d) Basidiospores, (e) Generative hyphae, scale bars: a=100 μm, b–e = 5 μm; (VII‒VIII) Fresh basidiomata of Auricularia nigricans, scale bars: 10 mm, (IX) Microscopic characters of Auricularia nigricans (a) Cross-section of basidioma , (b) Hairs, (c) Basidia, (d) Basidiospores, (e) Generative hyphae, scale bars: a=100 μm, b–e = 5 μm.
Basidiomata solitary to gregarious, mostly pileate, substipitate, gelatinous when fresh, rough coriaceous when dry, orbicular, 22–35 mm wide and 0.8–1 mm thick; abhymenial surface violet brown (10E6), densely pillose, margin entire; hymenium brownish red (10D6), smooth, lubricous.
Zona pilosa 146.54–318.94µm wide; hairs tufted 107.75–474.1 × 4.31–6.46µm, brown-hyaline, thick walled, gregarious, rounded tip. Zona compacta 17.24–34.48 µm wide, densely packed, individual hyphae indistinguishable. Zona subcompacta superior 21.55–64.45µm wide. Zona laxa superior 360.8–524.8µm wide, hyphae 3–4µm wide. Medulla 32.8–214.89µm wide, often pigmented, composed of a single band of hyphae, hyphae 2–3µm wide, oriented parallel to the surface. Zona laxa inferior 524.8–688.8µm wide, hyphae 3–4µm in diameter. Zona subcompacta inferior 90.3–210.7µm wide. Hymenium 64.65–107.75µm thick, pigmented, brown, clamp connection present. Basidia cylindrical, 30.17–34.48 × 4.3–5.1µm, transversely 3 septate. Basidiospores 9.48–12.54–13.79 × 4.31–4.43–4.74µm, cylindric˗allantoid, smooth walled, curved at tip, 1–2 oil droplets, Q = 2.2–3.2, Qm = 2.82.
Habit and habitat:
Solitary to gregarious, grown on dead wood of angiosperms.
INDIA, West Bengal, New Jalpaiguri District, Khairbari, 26.683º N 89.229º E, alt. 80m, K. Acharya. A. Roy and R. Saha, 27 July-2017, CUH AM308.
Auricularia nigricans (Sw.) Birkebak Looney and Sánchez- Garcia is well characterized by its densely pilose coriaceous brownish red (10D6) basidiocarp with smooth hymenial surface; brown hyaline hairs ranging from 107.75–474.1 × 4.31–6.46µm; presence of thick medullary layer 32.8–185µm and basidiospores measuring 9.48–13.79 × 4.31–4.74µm with a Qm value 2.82. The taxon has worldwide in distribution and previously reported from Brazil17, Columbia11 and Jamaica10. Our collection mostly matches with the specimens reported from Brazil with slight difference in length of zona pilosa (600–700µm vs 146.54–318.94 µm) and Columbia with slight variation in size of basidia (40–70µm vs 30.17–34.48µm). The description reported from Jamaica differs by having larger spores (12.5–15.5 × 6.5–7µm vs. 9.48–13.79 × 4.3–4.7µm). Among macro˗microscopically related taxa, Auricularia fuscosuccinea (Mont.) Henn. differs by having thin transparent basidiocarp, thin pilose zone (20–90µm) and smaller basidiospores (8–10 × 4.2–6.5µm)11; Auricularia cornea Ehrenb. differs by having larger spores (10.2–16.7 × 5–7µm) and thin medullary layer (75–125µm)18.
Turbinellus floccosus (Schwein.) Earle ex Giachini and Castellano Figs. 2(I‒III)
Basidiocarp pileate, infundibulliform, stipitate, 90–45 × 15–37mm in diam., reddish orange (7A8), typically funnel shaped. Hymenophore pseudolamellate, folded, white (1A1) to greyish red (7B3). Pseudolamellae dichotomously veined towards margin. Margin wavy, thick, concolourous with the pileus upper surface. Stipe short, 20–40 × 10–20mm in diam., distinct, cylindrical, slightly bulbous at base, orange red (8A8) when young and reddish golden brownish orange (6C4) at maturity.
Pileipellis cells 56.61–116.55 × 13.32–27.97µm in diam., thin walled, hyaline, flattened, broadly cylindricall in outline. Hyphal system monomitic; generative 2.99–9.99µm wide, thin walled, simple septate, sometimes with clamp, hyaline, branched. Pileus tramal gloeopherous hyphae 2.66–8.33µm wide, brownish orange (5C3), thin walled, branched. Stipe gloeopherous hyphae 1.67–6.66µm wide, brownish orange (5C3), thin walled, branched. Cystidia absent. Basidia 69.93–103.23 × 9.99–13.32µm in diam., clavate, hyaline, thin walled, long, 4-sterigmate. Basidiospores (12.99–)13.65–15.79–16.65(–17.32) × 6.33–6.74–7.33 µm in diam., broadly cylindrical, verrucose, thick walled, hyaline with brownish grey (5C2) deposites, Q=1.95–2.78, Qm=2.36.
Habit and habitat: Gregareous, grown on soil.
Specimen examined: INDIA, West Bengal, Darjeeling District, Mane bhanjang, 26.987⁰N 88.121⁰ E, alt. 1,928 m, S. Paloi, 17 March-2014, CUH AM276.
Turbinellus floccosus (Schwein.) Earle ex Giachini and Castellano is characterized by its pileate and stipitate basidiocarp; folded hymenophore, monomitic hyphal system; clamped generative hyphae; broadly cylindrical, verrucose, thick walled basidiospores measuring 12.99–17.32 × 6.33–7.33µm in diameter with mean Q value of 2.36. This taxon was previously reported from Uttarakhand, India and matches mostly with our specimen except in having slightly shorter basidia (50–80 µm vs 10–13µm) which may be attributed to the variation in climatic conditions12. Among macro-microscopically closely related species, Gomphus clavatus (Pers.) Gray differs by the presence of vinaceous brown to violet hymenium and Gomphus brunneus (Heinem.) Corner differs by the presence of smaller basidiospores (7.5–10 × 3.5–5μm vs 12.99–17.32 × 6.33–7.33µm)20.
Craterellus shoreae Hembrom, K. Das, A. Parihar and Buyck Figs. 2(IV‒VI)
Basidiomata 11–20 mm high, annual, terrestrial, small, gregarious, soft to more or less leathery, papery when dried. Pileus 7–16 mm diameter, 2–3 mm thick, slightly infundibuliform, nearly plane to uplifted with broadly undulating margin, centre deeply depressed, melon yellow (5A6). Margin entire, concolourous. Hymenophore smooth to wavy towards margin, covering entire underside of pileus. Stipe 9–18 mm long, 1–3 mm wide, cylindrical, melon yellow (5A6); Trama concolorous.
Pileipellis 114.8–163.59µm wide, composed of cylindrical hyphae, pale yellow in KOH; terminal cells 26.64–45.92 × 3.33–6.66µm, with distinct minute vesicle. Pileus tramal hyphae 5.74–8.61µm wide, pale yellow, branched, interwoven with distinct minute vesicle; cells 17.22–40.18 × 5.74–8.61 µm. Hymenium in transverse section 100.45–157.85 µm thick, pale yellow in KOH. Basidia 51.66–80.36 × 5.74–8.04µm in diameter, subcylindrical to subclavate, hyaline to pale yellow in KOH, thin walled; 4-sterigmate, sterigmata 5.74–14.35µm long, 0.57–5.17µm wide at base, acute pointed to tapered and few others curved. Cystidia absent. Cystidial element 48.79–111.93 × 5.74–12.05 µm wide, smooth, with numerous globose to irregular vesicle. Basidiospores 8.61–9.49–10.05(–11.48) × 5.17–5.81–6.31 µm in diameter, ellipsoid to elongate, smooth, thin walled with apiculae (0.3–0.9µm long), hyaline, with single to multiple yellowish guttules, Q=1.36–2, Qm=1.64. Stipitipellis composed of compactly arranged regular hyphae, thin walled, cells 26.64–39.96 × 3.9–4.9 µm and rounded at apex. Stipe trama composed of branched septate hyphae, hyaline in KOH, cells 23.31–39.96 × 2.66–3.99 µm. Clamp connections absent from all tissue.
Habit and habitat:
Gregarious, grown on soil under Shorea robusta Roth.
INDIA, West Bengal, New Jalpaiguri District, Apalchand forest, 26.7785° N 88.6434° E, alt. 84m, K. Acharya, A. Roy and R. Saha, 31 August-2018, CUH AM309.
Craterellus shoreae Hembrom, K. Das, A. Parihar and Buyckis well characterized by the presence of melon yellow colouredpileus, hymenophore smooth to wavy towards margin, presence of cystidial element, absence of clamp connection and basidiospores with single to multiple yellowish guttules, measuring 8.61–11.48 × 5.17–6.31µm with mean Q value of 1.64. We are reporting Clavaria zollingeri for the first time from West Bengal. Craterellus shoreae has been previously reported from Rajmahal hills of Jharkhand, India13. Considering the morphological features the description of our collected specimens mostly matches with earlier report from India, however varies a bit with regard to the larger size of basidia (36–58 × 6–11µm vs 51.66–80.36 × 5.74–8.04µm), sterigmata (3–7 × 1–2µm vs 5.74– 14.35 × 0.57–5.17µm), cystidial element (23–67 × 6–10 µm vs 48.79–111.93 × 5.74–12.05µm) that may be attributed to the reason of climatic and geographical variations. Among macro-microscopically related taxa, Craterellus albostrigosus C.K. Pradeep and K.B. Vrinda differs by having brownish gray to grayish brown cantharelloid basidiomata and 2–3–4–5 sterigmata; Craterellus inusitatus C.K. Pradeep and K.B. Vrinda differ by having chalky white pileus surface, cream or pale grayish towards base, absence of stipe or with a reduced minute eccentric to lateral pseudostipe21.
Clavaria zollingeri Lev. Figs. 2(VII‒VIII)
Fig. 2: (I‒II) Fresh basidiomata of Turbinellus floccosus, scale bars: 10 mm, (III) Microscopic characters of Turbinellus floccosus (a) Basidia (b) Pileipellis hyphae (c) Basidiospores (d) Genetative hyphae (e) Gloeopherous hyphae of pileus trama (f) Gloeopherous hyphae of stipe trama, scale bars: a–f = 5 μm; (IV‒V) Fresh basidiomata of Craterellus shoreae, scale bars: 10 mm, (VI) Microscopic characters of Craterellus shoreae (a) Basidia (b) Cystidial element (c) Genetative hyphae of pileus trama (d) Basidiospores (e) Genetative hyphae of stipe trama (f) Pileipellis hyphae (g) Stipitipellis hyphae. Scale bars: a–g = 5 μm; (VII) Fresh basidiomata of Clavaria zollingeri, scale bars: 10 mm, (VIII) Clavaria zollingeri (a) Basidia (b) Basidiospores (c) Generative hyphae with secondary septa. Scale bars: a–c = 5 μm.
Basidiomata 30–45 mm long, base up to 3‒4 mm wide, moderate in size, solitary, erect, branched, glabrous, greyish violet (18D4). Trunk sterile, glabrous, concolourous, brittle on drying. Branches dichotomous, divided up to 5 times, unequal, in alternating planes; primary branches flattened, up to 4 mm wide, ultimate branches radial, up to 1.2 mm wide; apices pastel violet (18A4), mostly rounded. No specific odour or taste. No change of colour in dilute alkali.
Hymenium amphigenous, only absent at the trunk. Hyphal system monomitic; generative hyphae 5.6–10.77µm wide, simple septate, branched, inflated, secondary septa present, clamp absent, smooth, hyaline, thin-walled, loosely interwoven. Basidia 21.55–38.79 × 4.74–8.62µm, elongated, narrowly clavate to clavate, light grey (19D1), multiguttulate, thin walled, 4-sterigmata; strerigmata straight, up to 1µm long. Basidiospores (4.74–)5.17−6.71−7.76 × 3.8–4.8–5.6(–6.03)µm, bluish grey (19D1), broadly ellipsoid, without ornamentation, smooth, thin-walled, aguttate, hilar apendix papillate, papilla less than 1 µm long, Q=1.09–1.8, Qm=1.3.
Habit and habitat:
Solitary, grown on humus.
INDIA, West Bengal, New Jalpaiguri District, Nagrakata, 26.880°N 88.907°E, alt. 214m, K. Acharya, A. Roy and R. Saha, 6 October-2018, CUH AM310.
Clavaria zollingeri Lev. possesses characteristic features like violet coloured fruit body, thin walled hyphae with secondary septa, absence of cystidia, absence of clamps, aguttate spores, multiguttulate basidia, basidiospores measuring 4.74–7.76 × 3.8–6.03µm with mean Q value of 1.3. Clavaria zollingeri is reported here for the first time from West Bengal. Previously this species has been reported from South Eastern Australia22, India (Mussoorie9 and Chamba12) and Malaya23.Our specimen nicely matches to the description previously reported from South East Australia. The specimen reported from Mussoorie and Chamba mostly matches with the present specimen except in having longer basidia (36–60 µm vs 21.55–38.79 µm, size of the basidia is same collected from Mussoorie and Chamba) of the both and smaller spore in case of chamba (4–6.5 × 3.5µm vs 4.74–7.76 × 3.8– 6.03µm) which may be attributed due to climatic variation. Among the macro and micro-morphologically related species, Clavaria indica Corner, K.S. Thind and Dev differs from Clavaria zollingeri on the basis of the presence of cystidia; Clavaria amoeinoides Corner, K.S. Thind and Anand and Clavaria fumosa Pers. differs due to the prensence of alutaceous and yellowish fruit body respectively9.
The authors would like to acknowledge Department of Environment, Government of West Bengal, India for providing financial assistance. The West Bengal Biodiversity Board and the Department of Botany (UGC-CAS Phase VII), University of Calcutta and DST-FIST are duly acknowledged for providing necessary infrastructural and administrative supports. Authors are grateful to directorate of forest, Government of West Bengal for extending support during the field visit.
CONFLICT OF INTEREST:
All authors have none to declare.
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Received on 28.05.2019 Modified on 25.06.2019
Accepted on 31.07.2019 © RJPT All right reserved