Aspergillus species isolated from mangrove forests in Borneo Island, Sarawak, Malaysia

 

JayaSeelan Sathiya Seelan 1, A.A.K. Faisal Ali 2 & Sepiah Muid 3

 

1 Institute for Tropical Biology and Conservation, Locked Bag 2073, UniversitiMalaysia Sabah, 88999 Kota Kinabalu, Sabah, Malaysia

2 Department of Biological Sciences, Texas Tech University, Lubbock, Texas 79409-3131, USA

3 Department of Plant Science and Environmental Ecology, Faculty of Resource Science and Technology, 94300 Kota Samarahan, UniversitiMalaysia Sarawak, Malaysia

Email: avinash80us@yahoo.com 1

 

 

Date of online publication 26 June 2009

ISSN 0974-7907 (online) | 0974-7893 (print)

 

Editor: V.B. Hosagoudar

 

Manuscript details:

Ms # o2192

Received 30 April 2009

Final received 03 June 2009

Finally accepted 17 June 2009

 

Citation: Seelan, J.S.S., A.K.K.F. Ali & S. Muid  (2009). Aspergillusspecies isolated from mangrove forests in Borneo Island, Sarawak, Malaysia. Journal of Threatened Taxa 1(6): 344-346.

 

Copyright: © Jaya Seelan Sathiya Seelan, A.K.K. Faisal Ali & Sepiah Muid 2009. Creative Commons Attribution 3.0 Unported License. JoTT allows unrestricted use of this article in any medium for non-profit purposes, reproduction and distribution by providing adequate credit to the authors and the source of publication.

 

Acknowledgements: Authors thank UniversitiMalaysia Sarawak (UNIMAS) for permission and MOSTI for financial support. They are grateful to Prof. Maren Klichof the United States Agricultural Department (USDA) for confirmation of identification.

 

 

For Images & Tables – Click here

 

The occurrence of Aspergillus is worldwide but mainly in tropical and subtropical regions (Domschet al. 1980; Christensen & Tuthill 1985). Aspergillus spp. areisolated from soil, sea water, plant debris, compost and litter (Raper & Fennell 1965). Aspergillusspp. are able to develop into many new strains through the process of heterokaryosis or parasexualreproduction and result high diversity in the environment. (Raper & Fennell, 1977; Papagianni2004). This study aims to project the diversity of Aspergillusspp. on mangrove soil and leaf litters of four mangrove locations in Sarawak.

 

Materials and Methods

Sample collection and fungal isolation: Mangrove soil and 100-200 g of dried leaf litters were collected at random from four locations, i.e. Sematan, Lundu,Kampung Bako, Bako in the Borneo Island of Sarawak.  Soil samples were pretreated following Warcup (1950).  Five replicates were made for different soil samples and incubated at 28°C for seven days. Dried leaf litter samples were sliced into 1cm  pieces,  directly plated onto Malt Extract Agar (MEA) and then incubated at 28°C for seven days.

Identification: For identification, the fungi isolates were grown on five different media such as Czapek Yeast Extract Agar incubated at 25°C (CYA25), CYA37 (incubated at 37°C), CY20S (added with 20% sucrose incubated at 25°C), Malt Extract Agar (MEA) and Czapek Doxsolution agar (CZ) (incubated at 25°C). For each culture, five plates were used in triplicates.  Each plate was inoculated at three points, equidistant from the center and incubated in the dark for seven days. The strains were identified using current universal keys of identification (Raper & Fennell 1977; Pitt 1979; Domschet al. 1980; Klich 2002).  Notes of IMI description were also used for identification.

 

Microscopic Observation

A small tuft of mycelium and conidiophores were lifted from a fairly young portion of the colony, placed in a drop of alcohol on a glass slide and gently teased out.  A drop of lactophenolblue or acid fuchsin was used to stain.  The appearance of foot cell, conidiophores, presence of metulae (sterigmata) and conidia were observed.  Digital photographs were taken by a Nikon camera attached to LeicaMicroscope.

 

Result and Discussion

Fungal Isolation: A total of 138 isolates belonging to five species of Aspergillus were isolated from the mangrove soil and leaf litter samples, collected from different sampling sites in four mangrove forests in Sarawak.  Based on both macroscopic and microscopic observations, using an identification key, individual isolates were classified within the genus Aspergillus, belonging to three subgenera, four section and five species. The fungal isolates were identified as A. terreus Thom, A.flavipes Bainier and Sartory, A. carneus Blochwitz, A. fumigatus Fresenius and A. clavatus Desmazieres.  The most frequently isolated species was A.flavipes (63.04%), followed by A. fumigatus (16.7%), A. terreus(13.04%), A. carneus (5.8%) and A. clavatus (1.44%). The total isolates of Aspergillus from mangrove soil and leaf litters in Sarawak shown in Figure 1.

 

Morphological Characteristics

All the five isolates showed significant growth rate level (P<0.05) on the media tested as in Table 1.  All the five species isolated were varying  intexture, vesicle shape, pigment production and sporulation(Images 1-5).

Klich (2002)  reported nearly 200 species of Aspergillus primarily from India, Europe, Egypt, Syria, tropical Africa, Japan and North America.  Since there is no record for Borneo Island, this work has documented five different species of Aspergillusfrom the mangrove soils and leaf litters. From this preliminary observation, A.fumigatus Fresenius, A. flavipes Bainier and Sartory, A.carneus Blochwitz and A.clavatus Desmazieres, were morphologically more variable in conidial ornamentation, phialide shapes and pigmentation than described by Raper & Fennell (1965). In this study, A. clavatus was found  interesting  because of its yellow pigment and long conidiophores ranging from 3-5 cm on the leaf litters found in Kampung Bako while the other isolate of A. clavatus from Sematan, had short conidiophores ranging from 2.5-3.5 cm. Besides that, A. carneus showed reddish sclerotia on MEA at room temperature.  This is an important character of this species.

 

Conclusion

In conclusion, five species of Aspergillus were obtained i.e. A. fumigatus Fresenius, A. flavipes Bainier and Sartory, A. carneus Blochwitz and A. clavatus Desmazieres are reported from mangrove soil and leaf litters at Kampung Bako, Bako, Lundu and Sematan, Sarawak. Morphological characteristics of these five species were distinct.  Colony growth characteristics were media dependent and were different for each species

 

References

 

Christensen, M. & D.E. Tuthill (1985). Aspergillus: an overview, pp. 195-209. In: Samson, R.A. & J.I. Pitt (eds.).Advances in Penicillium and Aspergillus systematics.New York: Plenum Press.

Domsch, K.H., W. Gams & T.H. Anderson (1980). Compendium of soil fungi. Academic Press, London. New York, Torroute, Sydney San Francisco, 859pp.

Klich, M.A. (2002). Identification of Common Aspergillusspecies. 1sted. Centraalbureau voor Schimmelcultures, Utrecht, Netherlands, 116pp.

Papagianni, M. (2004).Review of fungal morphology and metabolite production in submerged mycelial process. Biotechnology advances 22: 189-259.

Raper, K.B. & D.I. Fennell (1965). The genus Aspergillus. Williams and Wilkins, Baltimore, 686pp.

Raper, K.B. & D.I. Fennell (1977). The genus Aspergillus. Robert Erieger Publishing Company Huntington, New York, 68pp.

Warcup, J.H. (1950). The soil plate method for isolation of fungi from soil. Nature166: 117-118.