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PSRTAfUKA A Journal of Biological, Physical and Social Sciences

April 1992

Vol. 15 No. 1

Contents Section I: Biological Sciences Growth and Physiological Changes ofAverrhoa carambolaas Influenced 1>\ Water Availability- Mohd. Razi Ismail and Muhammad Awang Hosts of Pseudomonas solanacearum in Malaysia - //. Abdullah Kesan Sela Masa Pengawanan Semuladan Umur Sewaktu Pengasingan daripada Ibu ke atas Prestasi Reproduktifdan Tumbesaran Arnab New Zealand White - Chong, S.F., Hanafi bin Hanoi dan A.M. Majid

1 9 13

Communication Compositional Changes during Development and Maturation of Hydro ponically Grown Cucumber {Cucumis sativus I..) - II.M GhazaUandAbd. Gkam Hashim

21

A Life Table of the Asiatic Maize Stem Borer, Ostrinia furnacalu Guenee - M. Y. Hussein and MB. Ibrahim

27

Ketoksikan Beberapa Racun Hamama and Piretroid Tiruan terhadap Hamama Lelabah Merah, Tetranyckus urticae Koch Kompleks- Yusof Ibrahim, DzolkhiJU Omar and Adnan Othman Curcumin for Upgrading Skin Colour of Broilers ~ Awang I.P.R., U, ChulanandF.B.H. Ahmad

31 37

Section II: Physical Sciences Toxicity Studies of Plan! Extracts on Insects and Fish - Mohd. Aspollah Sukari, Mawardi Rah mam, Abd. Rahman Manas and Show Takahashi Effect of Carbon and Nitrogen Sources on the Growth and Production ofCellulase Enzymes of a Newly Isolated . 1 spergilius sp. - Chon>-(-hin I'ng and K. Rajendra Solvent Extraction of Cd(II) usingn-Butylamine-C10~-MIBKand n-Butylamine-Oxine-MIBK- W.T. TanandS.K Wong Calculation of the Effective Stopping Power of Ions Generated by Neutrons in Tissue Constituents - EUas Saion and O.K. Watt Mass Spectrometric Studies of Positive Ion Molecule Reactions in \ H and SV Gases- ZainalAbidin Talib & U Saporoschenka Kegunaan Sub-Martingale dalam Masalah Storan - Mohd. Kidin Shah ran

\1

15 51 55 61 69

Communica tion FI-IR Spectroscopic Studies on Lignin from Some Tropical Woods and Rattan - \. I.an cr Ramli Ibrahim Section III: Social Sciences January Effect on the Thinly Traded KLSE: Tests with Appropriate Refinements - M.N. Annuar, M, Ariff and M. Sham she)

85

A Scientific Journal published bj

UNIVERSITI PERTANIAN MALAYSIA

ISSN 0126-6128

EDITORIAL BOARD

CHIN HOONG FONG (Chief Editor)

FATIMAII MO1H). ARSIIAD (Business Manager) SULAIMAN MOHD \ASSIN ANG KOKJEE MOHD. ZOHADIE BARDAIE MAT YUSOFF ABDULIAH

SUHAIIA MOHAMED VIDYADARAN MENON KAREN ANNE CROUSE BADRI AHMAD SAID SAJAP ABDUL. RAHMAN MOHD AROFF

SUMANGALA PILLAI (Secretary) PERTANIKA is a scientific journal published thrice a year (April, August and December) by Universiti Pertanian Malaysia (University of Agriculture, Malaysia) in which papers in Bahasa Malaysia and English in any area aligned with the work done at the faculties of the University appear. Currently these include Agriculture, Forestry, Veterinary and Animal Science, Food Science and Technology, Economics and Management, Engineering, Fisheries and Marine Science, Science and Environmental Studies, Extension and Continuing Education, Education and Social Studies, Human Development and Consumer Studies. PERTANIKA welcomes original reports in English or Bahasa Malaysia of research not previously or simultaneously published in any scientific or technical journal from the staff of Universiti Pertanian Malaysia and other local and overseas institutions and organisations. Contributions are reviewed by a panel of consultants whose names appear in the last issue of each volume. PERTANIKA is currently abstracted by the following; Agrindex, Biological Abstracts, Chemical Abstract, Nutrition Abstracts, Animal Breeding Abstracts, Field Crop Abstracts, Forestry Abstracts, Forest Production Abstracts, Herbal Abstracts, Horticultural Abstracts, Indian Veterinarian, Plant Breeding Abstracts, Reviews in Applied Entomology, Reviews in Plant Pathology, Soils and Fertilizers and Veterinary Bulletin. Articles in triplicate should be submitted to the Chief Editor, PERTANIKA,Universiti Pertanian Malaysia, Serdang, Selangor, Malaysia. Subscription Rates(per year) Malaysia/Singapore

Overseas

Individual

$45,000

US$30,00

Institutions

$70.00

US$35.00

Oversea subscribers - US $6.00 per issue for airmail surcharge. Cheques/Bank drafts should be made payable to UNIVERSITI PERTANIAN MALAYSIA and sent to, PERTANIKA, Universiti Pertanian Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.

OFFICE *FKocJO€ATo»« Uf*M"MTQC> Technology Cu

A Journal of Biological, Physical and Social Sciences Number 1, April 1992 Contents Section I: Biological Sciences Growth and Physiological Changes of Averrhoa carambola as Influenced by Water Availability - Mohd. Razilsmall and Mu ham mad A wa ng Hosts of Pseudomonas solanacearum in Malaysia - H. Abdullah Kesan Sela Masa Pengawanan Semula dan Umur Sewaktu Pengasingan daripada Ibu ke atas Prestasi Reproduktif dan Tumbesaran Arnab New Zealand White - Chong, S.F., Hanafi bin Harun dan AM. Majid

1 9 13

Communication Compositional Changes during Development and Maturation of Hydroponically Grown Cucumber (Cucumis sativus L.) - HM. Ghazali and Abd. Ghani Hashim A Life Table of the Asiatic Maize Stem Borer, Ostrinia furnacalis Guenee - M. Y. Hussein and M.B. Ibrahim

21 27

Ketoksikan Beberapa Racun Hamama and Piretroid Tiruan terhadap Hamama Lelabah Merah, Tetranychus urticae Koch Kompleks - Yusof Ibrahim, Dzolkhifli Omar and Adnan Othman Curcumin for Upgrading Skin Colour of Broilers- Awangl.P.R., U. Chulan andF.B.H. Ahmad* Section II: Physical Sciences Toxicicy Studies of Plant Extracts on Insects and Fish - Mohd. Aspollah Sukari, Maiuardi Rahmani, Abd. Rahman Manas and Shozo Takahashi Effect of Carbon and Nitrogen Sources on the Growth and Production of Cellulase Enzymes of a Newly Isolated Aspergillussp, - Chozo-Chin Tong and K. Rajendra Solvent Extraction of Cd(H) using n-Butylamine-ClO^-MIBKandn-Butylamine-Oxine-MIBK-W.7. TanandS.K. Wong Calculation of the Effective Stopping Power of Ions Generated by Neutrons in Tissue Constituents - Elias Saion and O.K. Watt Mass Spectrometric Studies of Positive Ion/Molecule Reactions in NH^ and SF6 Gases - Zainal Abidin Talib 6f M. Saporoschenko Kegunaan Sub-Martingale dalam Masalah Storan - Mohd. Kidin Shahran

31 37

41

45 51 55 61 69

Communication FT-IR Spectroscopic Studies on Lignin from Some Tropical Woods and Rattan - S. IMU 6f Ramli Ibrahim

75

Section III: Social Sciences January Effect on the Thinly Traded KJLSE: Tests with Appropriate Refinements - M.N. Annuar, M. Ariff and M. Shamsher

85

Members of the Editorial Board Prof. Chin Hoong Fong M. Agric. Sc, Ph.D. (Melb.) F.I. Biol. (Land).

Faculty of Agriculture (Chief Editor)

Prof. Sulaiman bin Mohd Yassin B. Agric. Sc. (Hons.) Malaya, M.RA. (C.A.), Ph.D. (Cornell).

Centre for Extension and Continuing Education

Prof. Ang Kok Jee B. Sc. (Madras), M. Sc. (Malaya), Ph.D. (Waterloo)

Faculty of Fisheries and Marine Science

Faculty of Engineering Assoc. Prof. Mohd Zohadie bin Bardaie Dip. Agric. (Malaya), B.S., M.S. (U.C. Davis), Ph. D. (Cornell) Assoc. Prof. Mat Yusoff Abdullah B. Sc. (Malaya), M.S. (U.P.L.B.), Ph.D. (Okla. State)

Faculty of Science and Environmental Studies

Ahmad Said bin Sajap Dip. Agric. (Malaya), B. Sc. (Calif.), M.S. (Iowa State), Ph.D. (Iowa State)

Faculty of Forestry

Assoc. Prof. Suhaila Mohamed B.S. (Leeds), Ph.D. (Leeds)

Faculty of Food Science and Biotechnology

Fatimah Mohd. Arshad B. Ec. (Hons) (Malaya), M. Sc, Ph.D. (N'cle) (UK)

Faculty of Resource Economics and Management (Business Manager)

Assoc. Prof. Vidyadaran Menon D.V.M. (Mymensingh), M.V.S. (Melbourne), Ph.D (UPM)

Faculty of Veterinary Medicine and Animal Science

Assoc. Prof. Karen Anne Crouse-Badri B.Sc (Hons), (St. F.X.) M. Sc. (Dal.) Ph.D. (Malaya)

Faculty of Science and Environmental Studies

Assoc. Prof Abdul Rahman bin Md. Aroff B.A. Dip. Ed. (Malaya), M. Ed. (Nheff), Ph.D. (Southampton)

Faculty of Educational Studies

Sumangala Pillai B. Soc Sc. (Hons)

Universiti Pertanian Malaysia Press (Secretary)

Section I Biological Sciences

Pertanikal5(l), 1-7 (1992)

Growth and Physiological Changes of Averrhoa carambola as Influenced by Water Availability MOHD. RAZI ISMAIL1 and MUHAMAD AWANG2 Department of Agronomy and Horticulture, department of Environmental Science Universiti Pertanian Malaysia, 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia.

1

Key words: Water stress, Averrhoa carambola^ growth, leaf water potential, photosynthesis rate, stomatal conductance. ABSTRAK Kajian tegasan air ke alas pertumbuhan dan fisiologi yang berkaitan bagi tanaman Averrhoa carambola dijalankan dalam dua kajian berasingan menggunakan kaedah gabungan tegasan air. Kajian pertama jelas menunjukkan pengurangan pertumbuhan berkait rapat dengan pengurangan kedapatan air. Terdapat korelasi yang signifikan di antara potensi air dan pengurangan kedapatan air. Terdapat korelasi yang signifikan di antara potensi air dan pengurangan konduksi stomata, kadar transpirasi dan kadar fotosintesis. Pengurangan kadar fotosintesis hanya berlaku apabila potensi air daun mengurang sehingga -0.85 MPa. Walau bagaimanapun, pengurangan kandungan klorofil hanya berlaku pada tahap tegasan air yang ketara. Kaitan di antara ciri-ciri fisiologi dan pertumbuhan vegetatif di bincangkan. ABSTRACT Utilizing an integrated soil moisture stress approach, two different experiments were conducted simultaneously to investigate the effects of water stress on growth and related physiological characteristics J)f Averrhoa carambola. The first experiment clearly indicated a high correlation between soil water availability and a reduction in plant vegetative grozvth. In the second experiment, there was a significant correlation between leaf water potential and a reduction in stomatal conductance, transpiration rate and photosynthesis rate. The inhibition of photosynthesis rate was only apparent when leaf water potential was reduced to -0.85 MPa, However, chlorophyll content was only affected by a further reduction in water availability. The relationship between physiological characteristics and vegetative growth is discussed. INTRODUCTION Starfruit (Averrhoa carambola) has been recognized as one of the major crops in the Malaysian fruit industry. According to the Department of Agriculture Malaysia (DOA, 1990), establishment of young plants in the field must be carried out during the early rainy season to ensure better root growth. In addition, Coloma (1972) suggested that irrigation should be carried out for the first 2 - 3 years in areas with a prolonged dry season. However, information relating to the effects of water availability on growth and its physiological responses are generally scarce and details of the physiological water status pertaining to this particular crop need further investigation.

It is acknowledged that water status could limit growth and plant development through its effect on plant physiological processes (Kramer 1983; Jones 1990. The responses of plant physiological processes also vary according to the status of water potential developed in a plant. Basically, water stress may reduce leaf water potential which consequently reduces turgor potential in the cell which in turn affects cell expansion (Hsiao and Acevedo 1974). Considering the importance of water requirements of this crop, this study was undertaken to examine the growth and plant physiological characteristics that are influenced by differing water availability. The study also aimed at elucidating the relationship between physiological responses and water status in the plants.

MOHD. RAZI ISMAIL AND MUHAMAD AWANG

MATERIALS AND METHODS

The experiments were conducted at the Greenhouse Unit at Universiti Pertanian Malaysia, Serdang, Selangor. The average daily temperature ranged between 28-35°C and the relative humidity was between 65-72%. Four-month-old uniform plants of Averrhoa carambola of clone B17 grafted on BIO stock plants were planted in pots which were 38 x 30 x 28 cm in size containing 13 kg soil mixture of 3:2:1 (top soil: organic manure: sand). Plants were raised according to the recommendations of DOA (1990). Two different sets of experiments were conducted simultaneously. In the first experiment, plants were watered to field capacity at intervals of 0, 3 and 6 days for 12 weeks, whilst in the second set of experiments, plants were subjected to water stress for 3, 7, 10, 14, 17 and 21 days. The changes in plant physiological characteristics were monitored for the period of water stress. Both experiments were conducted in a completely randomized design with 4 replications. Stem extension rate was recorded at weekly intervals. After twelve weeks of treatment, leaves were enclosed in the polythene bags and individual leaf area was recorded using an automatic leaf area meter (Delta-T, Cambridge, UK). Plants were separated into shoots and roots and oven dried at 80°C for 48 hours. Trifoliate terminal leaflets with 2 cm petiole were excised and immediately placed in a pressure chamber (PMS Ins. Utah, U.S.A) to determine leaf water potential. Techniques and precautions for the determination were as described by Turner (1981). Stomatal conductance and transpiration rate were determined using a steady state porometer (Ll-1600 Li Cor Inc. Nebraska, U.S.A). Measurements were conducted between 1100-1230 h. The measurements of photosynthesis and respiration rates were based on polarographic methodology (Delliu and Walker 1972; Saka and Chisaka 1985). Chlorophyll content was determined according to Mackinney (1942). Measurements of these plant processes were carried out at the vegetative growth stage of the plants. RESULTS

Fig. 1 illustrates stem extension as influenced by water stress. The stem elongation of plants irrigated at 6-day intervals was reduced to 56% compared to control at termination. However, there was no significant difference (P>0.05) between the control and the plants irrigated at 3-day intervals within a week. A similar trend was observed in leaf 2

area and its biomass (Table 1). Reduction in leaf area was 15% and 47% as compared to the control when plants were subjected to water stress for 3 and 6 days respectively. Contrary to expectations, there was no significant difference (P>0.05) between control and 3 days of water stress on root biomass. However, the root:shoot ratio was significantly higher (P>0.05) on plants treated with 3 days of water stress as compared to the control and 6 days of water stress. There was a general reduction in all physiological processes as a result of a reduction in a leaf water potential. Inhibition of photosynthetic O2 evolution and respiratory activities was accompanied by a decrease in stomatal conductance resulting in a lower transpiration rate. The compensation point for assimilation of CO2 was only achieved after 22 days of water stress as indicated in Fig. 2. The results also revealed that the photosynthesis rate was significantly reduced when the leaf water potential dropped to - 0.85 MPa after day 7, suggesting that plant photosynthesis is sensitive to small changes in water stress. Correlations between rates of photosynthesis (r=0.89) and transpiration (r=0.92) with respect to changes in leaf water potential are presented in Figures 3 and 4 respectively. These linear responses of leaf photosynthesis and transpiration rates to water stress reflect the behaviour of stomatal functioning in contributing to gas exchange and efficiency of water utilisation. Furthermore, a close relationship between photosynthesis rates and stomatal conductance (Figure 5) along with the water witholding process was exhibited by the treated plant. In contrast, chlorophyll content was only significantly reduced after 7 days of water stress which coincided with the reduction of photosynthesis rate as high as 33% compared to day 1. DISCUSSION The most pronounced effect of reduction of water availability to the plants was growth inhibition (Begg and Turner 1976; Takami et ai 1981; Li et al. 1989). Cell enlargement is one of the most sensitive processes affected by a change in plant water status (Hsiao 1973). The relationship between growth reduction and plant water status has been widely reviewed elsewhere (Kramer 1983; Schulze 1986). When plants were grown at 3-day intervals of water stress, root:shoot ratio increased. The change in root: shoot ratio is generally considered an adaptive mechanism of plants exposed to mild water stress. Begg and Turner (1976) suggested that a greater allocation of the limited

PERTANIKAVOL. 15 NO. 1, 1992

GROWTH AND PHYSIOLOGICAL CHANGES OF AVERRHOA CARAMBOIA

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Weeks from start of treatments Fig. 1:

Stem elongation (cm) as influenced by different water availability. Tl = daily irrigation; T2 = irrigation at 3 day intervals; 13 = irrigation at 6 day intervals. Bars represent LSD 5%.

TABLE 1 Plant vegetative growth as influenced by different water availability. T l : daily irrigation; T2 : 3 day interval irrigation; T3; 6 day interval irrigation Treat.

Leaf area (cm)

Tl T2 T3

1,805%

Leaf Dry Wt (g/plant)

Stem Dry Wt (g/plant)

Root Dry WT (g/plant)

Root: shoot ratio

960.5 817.4 626.0

6.5 4.6 3.4

24.6 15.0 8,0

27.8 26.8 14.4

4.3 5.8 4.2

112.7

0.5

4.4

4.7

0.9

PERTANIKA VOL. 15 NO. 1, 1992

MOHD. RAZI ISMAIL AND MUHAMAD AWANG

30(

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20-

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r = 0.98 **

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IXiration of witholding water (Days)

Fig1. 2: Relationship between photosynthesis rate, transpiration rate and duration ofxoitholding water. Lines are fitted roith a linearregression. Q=Photosynthesis rate;% ^respiration rate; ** - significant at P = 0.01

-3.0

-2.0

-1.5

-1.0

-0.5

Leaf water potential (MPa) Fig. 3: Relationship between leaf water potential and photosynthetic rate. ** = significant at P = 0,01

PERTANIKAVOL. 15 NO. 1, 1992

GROWTH AND PH\SIOLOGICAL CHANGES OF AVERRHOA CARAMBOIA

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35

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25 20

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PERTANIKA VOL. 15 NO. 1, 1992

MOHD. RAZI ISMAIL AND MUHAMAD AWANG

TABLE 2 Leaf water potential (Uw), Photosynthesis rate (Pn), Respiration rate (Rs), Chlorophyll content (Chi), Transpiration rate (Tr) and Stomatal conductance (gs) as influenced by duration of water stress Days

Uw

(MPa) 0 3

7

-0.7 -0.7 -0.9

10

-1.3

14

-2.0 -2.4 -2.9

17 21

LSD5%

0.2

Pn

30.3 27.6 20.2 17.1 12.8

(mg/dnr )

(sec cm )

gs (mm s"1)

95.0 91.4 89.4 86.7 68.3 47.8 34.8

9.09 9.11 6.34 6.25 5.26 4.00 2.38

8.7

0.04

1

11.5

8.3

5.4

6.1

4.6

3.35 3.34 3.36 2.76 2.65 2.67 2.65

3.1

1.4

0.48

11.4 10.0

carbon is available to the root than to shoot when there is a water deficit in plants. The results also indicated that a small change in plant water status results in a reduction in most of the plant physiological parameters studied (Table 2 and Figure 3). The present studies show that leaf photosynthesis rate is affected by water potential greater than 0.85 MPa. The response of the photosynthesis rate to a small change in leaf water potential agrees with the findings of Schulze (1986) who suggests that a reduction in the photosynthetic rate when water deficit develops arises from the gradient of water potential that develops locally within the leaf tissue between xylem and the site of evaporation. In Zea mays, Barlow et al (1977) showed that leaf photosynthesis was affected when the reduction of water potential in plants was greater than - 1.2MPa. They also showed that stomatal conductance and transpiration rates were reduced with the changes in plant water status. This agrees with the findings of Aston (1973) and Sheriff (1984). Bradford and Hsiao (1982) suggest that the decline in respiration rate during water stress is related to a reduction in water demand as growth is inhibited. CONCLUSION This study indicates the importance of adequate water for the early establishment of Averrhoa carambola in the field. The reduction in growth when a water deficit develops is associated with changes in the plant physiological processes such as stomatal conductance and photosynthesis rate. Detailed studies need to be undertaken to further understand the mechanism of stomatal response

Tr

Chi

Rs (umol02dm"*h"»)

6.5 6.1

1

affecting photosynthesis and osmotic adjustment in the plant. ACKNOWLEDGEMENTS

The authors would like to thank the IRPA Fruit Group of the Faculty of Agriculture, and Faculty of Science and Environmental Studies, University Pertanian Malaysia for providing a grant for this study. We thank Mr. Syed Razlan and Mr. Kamaludin Mahmod for their technical assistance. REFERENCES

MJ. 1973. Changes in Internal Water Status and the Gas Exchange of Leaves in Response to Ambient Evaporative Demand, In Plant Response to Climatic Factor ed. R.O. Stayter, Proceedings Uppsala Symposium (1970). UNESCO p. 243-247.

ASTON,

E.W.R., L. BOERSMA, and J.L. YOUNG. 1977. Photosynthesis, Transpiration Rate and Leaf Elongation in Corn Seedlings of Suboptimal Soil Temperatures. Agron. J. 69: 95-100.

BARLOW,

BEGG, J.E.

and N.C. TURNER. 1976. Crop water deficit. Adv. Agron. 28:161-217. K.J. and T.C. HSIAO. 1982. Physiological Responses to Moderate Water Stress. In: Encyclopedia of Plant Physiology. Water Relation and Carbon Assimilation eds. O.L. Long, D.S. Nobel, C.B. Osmond and H. Ziegler 264-324 Berlin, Tokyo: Springer-verlag.

BRADFORD,

G.R. 1972. Kamias Averrhoa bilimbi L., (Oxalidaceae). In Cultural Direction for Philippine Agricultural Crops. Vol. I (Fruits) p. 142145.

COLOMA,

PERTANIKAVOL. 15 NO. 1, 1992

GROWTH AND PHYSIOLOGICAL CHANGES OF AVERRHOA CARAMBOIJK

DELLIU, T J and D.A. WALKER 1972. An Improved

Method for the Measurement of Photosynthesis Oxygen Evolution by Isolated Chloroplast. New PhytoL 71: 201-225. DEPARTMENT OF AGRICULTURE, MAIAYSIA. 1990. Belimbing

BesL Booklet Number 28: 24 pp.

MACKINNEY, G. 1942 Absorption of Light by Chlorophyll Solutions./ Biol Chemistry 140: 315-322. SAKA, E and H. CHISAKA. 1985. Photosynthetic Meas-

urement by Oxygen Electrode as a Simple Bioassay Method. Jap. Agric. Research Quart. 18(4): 252-259.

HSIAO, T.C. 1973. Plant Responses to Water Stress. Ann. Rev, Plant Physiol. 24: 519-570.

SCHULZE, E.D. 1986. Whole Plant Response to Drought. Aust.J. Plant Physiol 13: 127-141.

HSIAO, T.C. & E. ACEVEDO. 1974. Plant Responses to

SHERIFF, W.D. 1984. Epidermal Transpiration and Stomatal Responses to Humidity : Some Hypothesis Explored. Plant, Cell and Environ. 7: 669677.

Water Deficits, Water Use Efficiency and Drought Resistance. Agric Meteorology 14: 59-84. JONES, H.G. 1990. Physiological Aspects of the Control of Water Status in Horticultural Crops. Hort. S « 2 5 ( l ) : 19-24. KRAMER, P.J. 1983. Water Relations of Plants. Academy Press, New York, 481 p. Li, S.H., J.G. HUGUET, P.G SCHOCH and P. ORLANDO.

1989. Response of Peach Tree Growth and Cropping to Soil Water Deficit at Various Phenological Stages of Fruit Development. / . Hort. Sci. 64: 541-552.

TAKAMI, S., N.C. TURNER and H.M. RAWSON 1981. Leaf

Expansion of Four Sunflower (Helianthus annuus L.) Cultivars in Relation to Water Deficits. 1 Patterns during Plant Development. Plant, Cell and Environ. 4: 399-407. TURNER, A.C. 1981. Techniques and Experimental Approaches for the Measurements of Plant Water Status. Plant and Soil 58: 339-366.

PERTANIKA VOL. 15 NO. 1, 1992

(Received 8 January 1992)

Pertanika 15(1), 9-12 (1992)

Hosts of Pseudomonas solanacearum in Malaysia H. ABDULLAH Department of Plant Protection, Faculty of Agriculture, Universiti Pertanian Malaysia, 43400 UPM, Serdang, Selangor Darul Ehsan, Malaysia. Key words: Pseudomonas solanacearum^ host, Malaysia, tomato. ABSTRAK Satu kajian telah dijalankan untuk mengenal pasti perumah-perumah bagi Pseudomonas solanacearum di Malaysia. Kajian ini menunjukkan bahawa patogen ini didapati tersebar luas. Patogen telah di asingkan daripada 25 spesies pokok-fokok tanaman, rumpai dan perhiasan yang berpenyakit. Ujian inokulasi rumah kaca menunjukkan bahawa selain perumah asalnya, semua strain-strain patogen ini juga didapati patogenik kepada tomato. ABSTRACT A survey was conducted to determine the hosts of Pseudomonas solanacearum in Malaysia. The study showed that the pathogen was widespread for the pathogen was isolated from 25 species of diseased crop plants, weeds and ornamentals from various localities. Greenhouse inoculation tests showed that, besides their respective hosts, all isolates of the pathogen were also pathogenic to tomato. INTRODUCTION Bacterial wilt caused by Pseudomonas solanacearum

is widely distributed especially in the tropical regions of the world. Buddenhagen and Kelman (1964) considered it to be one of the most important, widespread and lethal bacterial diseases of plants. In Malaysia, bacterial wilt was first reported on potato and tomato in 1910 (Bancroft. 1910). Since then, its occurrence on a wide range of hosts has been reported by several workers (Burnett 1949; Thompson and Johnston 1953; Johnston 1960; Singh 1973: Abdullah 1980 a and b, 1983 a and b). In Malaysia, estimates of losses due to the disease are not available. However, on tomato, Graham et al. (1977) considered bacterial wilt to be the most serious factor limiting commercial production in the lowlands. The disease has resulted in the abandonment and discontinuation of the growing of tomato and ginger in severely affected areas. Resistant varieties developed, such as in the case of tomato, did not remain resistant in the tropics possibly due to conditions conducive to severe disease development in the tropics and/or to the existence of different strains of the pathogen. Thus, until adequate resistance can be

achieved, the control of the disease by other means, such as crop rotation and manipulation of cultural practices, seems to be the best solution to the problem. However, since P. solanacearum infects a wide range of cultivated and weed hosts, it is essential that these hosts of the local strains of the pathogen are known before the implementation of any crop rotation programme in a particular area. Therefore, in this study, attempts were made to determine the distribution of bacterial wilt in relation to its natural hosts and the pathogenicity of the isolates to its respective hosts and/or on tomato (Lycopersicon esculentum Mill). This infor-

mation is important for the formulation of effective control measures involving crop rotation and appropriate cultural management for the control of bacterial wilt of tomato in Malaysia. MATERIALS AND METHODS

The locations surveyed for bacterial wilt in this study are indicated in Figure 1. Plants were observed for typical bacterial wilt symptoms, viz. wilting of the plants (with or without yellowing of the leaves). The suspected plants were examined for exudation of bacterial ooze by placing the cut end of the stem in water. Positive diagnoses were

H. ABDULLAH

made when fine-milky-white strands of bacterial masses oozed out from the vascular tissues of the cut end of the stem after a few minutes. Infected hosts were also collected for isolation of the pathogen. In most cases, the pathogen was isolated from the stem except in older plants when isolation was also made from roots. In the case of ginger and dahlia, isolations were also made from the rhizomes and the tuberous roots respectively. The section of the plant used for isolation was washed in running water and dried with clean paper towels. The epidermis was removed aseptically and a segment approximately 3 - 5 mm in length was placed in ca. 5 ml of sterile distilled water for 20 - 30 min. A loopful of the bacterial suspension was then streaked on triphenyl tetrazolium chloride (TZC) medium (Kelman 1954) plates and incubated for 40 - 48 h at 30 ± 1°C. Fluidal white colonies with or without red centres that are typical of P. solanacearum appearance on TZC were selected and restreaked on TZC plates twice or until the colonies appeared pure. Isolates were then streaked on sucrose-peptone (SP) medium (Hayward 1960) or Casamino-acid peptone glucose (CPG) medium (Cuppels et al 1978) and incubated for 48 - 72 h. at 30 ± 1°C prior to preparation of stock culture. Stock cultures were maintained as bacterial suspension in ca. 5 ml sterile tap water and stored at 20 ± 1°C for further studies. Identification of the isolates was based on comparison of cultural and biochemical characteristics (Hayward 1964) with those of the confirmed isolates of P. solanacearum (Abdullah 1980 a and b). Pathogenicity tests were performed by stem pinprick inoculation and root inoculation of the respective hosts and tomato (Abdullah 1988). Inoculation of tomato was to determine whether isolates obtained from other hosts could also infect tomato. Control plants were similarly inoculated for each of the techniques but using sterile distilled water.

ing on land left fallow after a crop or among crop plants or even on roadsides. In most instances, whenever the infected weeds were found among susceptible crops, the crops were also observed to be infected with bacterial wilt. TABLE 1 Natural hosts of Pseudomonas solanacearum recorded

Host

Family

No. of

Types of

isolate plant Ageratum conyzoides L. Compositae Arachis hypogaea L. Leguminoseae Capsicum annum L. Solanaceae Capsicum grossum L. Solanaceae Ckome speciosissima Deppe. Capparidaceae Corchorus oliiarius L. Tiliaceae Cosmos caudatus Cav. Compositae Croton hirtus V Herit. Euphorbiaceae Dahlia rosea Cav. Compositae Euphorbia hiria L. Euphorbiaceae Euphorbia prunifolia Jacq (. Euphorbiaceae Hyptis capitata Jacq. Labiatae Ipomoea setosa L. Convolvulaceae Ixonurus sibiricus L. Labiatae Lycopersicon esculentum Mill. Solanaceae Nicotiana tabacum L. Solanaceae Phaseolus vulgaris L. Leguminoseae Physalis minima L. Solanaceae Psophocarpus tetragonolobus DC. Leguminoseae Solarium melongena L. Solanaceae Solanum tuberosum L. Solanaceae Synedrella nodiflora Gaertn. Compositae Tagetes erecta L. Compositae Zingiber officinale Rose. Zingiberaceae Zinnia elegans Jacq. Compositae

8

W*

26 22 2

c c c

5

w w c w o w w w o c

3 2 22 8 2 7 6 4 3 20 21 11 2

c c c w

6 18 3

c c c

4 6 29 3

w o c o

C = Crop plants. W = Weeds. O = Ornamentals.

RESULTS AND DISCUSSION

Species of natural hosts of P. solanacearum recorded in this study are presented in Table 1. A total of 243 isolates were obtained from 25 species of crop plants, weeds and ornamentals belonging to 9 families. Bacterial wilt was observed in all the areas sampled (Figure 1). Bacterial wilt was observed and the pathogen isolated throughout the year at the farm of the Universiti Pertanian Malaysia CUPM). Besides crop plants, the disease was observed commonly infecting weeds found grow10

Isolates were aerobic Gram-negative rods, oxidase and catalase positive, able to reduce nitrate to nitrite but did not produce fluorescent pigment on King's medium B (King et al 1954). Their cultural and biochemical characteristics were similar to those of P, solanacearum and were therefore identified as such. All the isolates collected during the earlier part of the study were subsequently confirmed by the Commonwealth Mycological Institute. The pathogenicity of all

PERTANIKAVOL. 15 NO. 1, 1992

HOSTS OF PSEUDOMONAS SOLANACEARUM IN MALAYSIA

ble to the local strains of P. solanacearum, such as in the case of winged bean (Psophocarpus tetragonolobus L.) DC. (Abdullah 1980a). Out of the 25 hosts of the pathogen that were recorded, 10 were common weeds. These weed species could serve as reservoir hosts during the absence of crop plants. Thus, it is essential that such alternate hosts be identified especially for successful implementation of crop rotation programmes designed to control bacterial wilt. It is interesting to note that bacterial wilt of sweet potato (Ipomoea batatas) and cassava {Manihot

esculenta) has, so far, not been reported in Malaysia. The former has been reported in China (He et al. 1983) and the latter in Indonesia (Nishiyama et al. 1980). The absence of the disease in these commonly grown crops in Malaysia places greater emphasis on the need for stricter quarantine measures to prevent the introduction of exotic strains into this country. CONCLUSION 111°

113°

115°

11?

IIS £

Fig 1: Locations of the areas sampled for bacterial wilt caused

by Pseudomonas solanacearum, isolates was confirmed on its respective host and tomato by both the stem and root inoculation techniques and the pathogen reisolated from the inoculated plants. Since all isolates were pathogenic to tomato, these hosts, whether crop plants or otherwise, could act as alternate hosts of the pathogen whenever tomato are not cultivated. Thus, future breeding programmes and varietal resistance studies designed to control bacterial wilt should take into consideration the many strains of the pathogen and its many alternate hosts. Control of weeds which are hosts of the pathogen may also be essential for successful crop rotation programmes. Results of this study showed bacterial wilt to be widespread. Even though the disease was observed on 25 plant species it was not observed on a number of hosts previously reported in Malaysia,

Bacterial wilt caused by P. solanacearum is widespread in Malaysia. The existence of the many weed hosts of the pathogen which are also pathogenic to tomato, indicates that for successful implementation of crop rotation programmes, designed to control bacterial wilt of tomato, it is essential that weeds be controlled. ACKNOWLEDGEMENTS

The author wishes to thank Universiti Pertanian Malaysia (UPM) for the research grant that made this study possible. Thanks are also due to all personnel from the Department of Agriculture Malaysia and the Malaysian Agricultural Research Development Institute for their invaluable help during the survey and to the laboratory staff of the Department of Plant Protection (UPM) for their help in the laboratory and glasshouse. Last but not least, I wish to thank the Commonwealth Mycological Institute (C.A.B. International Mycological Institute), Kew, England for the confirmation of some of the isolates. REFERENCES

such as Brassica chinensis, Vigna sinensis (L) EndL

ex. Hassk. (Singh 1973) and Amaranthus sp. (Thompson and Johnston 1953). Since the above crop plants were commonly seen growing in soils known to be infested with P. solanacearum such as at the farm of the UPM, this indicates the possible varietal resistance of the local varieties. Introduced varieties have been known to be suscepti-

ABDULLAH, H. 1980a. A Disease of Winged Bean (Psophocarpus tetragonolobus) Caused by Pseudomonas solanacearum in Malaysia. Plant Dis. 64: 798-799. ABDULLAH, H. 1980b. Pseudomonas solanacearum isolated from Two New Weed Hosts. FAO Plant Protection Bull 28: 79-81.

PERTANIKA VOL. 15 NO. I, 1992

H. ABDULLAH ABDULLAH, H. 1983a. Record of Additional New Hosts of Bacterial Wilt Pathogen (Pseudomonas solanacearum) in Malaysia. Malaysian Appl Bioi 12(1): 59-60. ABDULIAH, H. 1983b. Distribution of Bacterial Wilt Caused by Pseudomonas solanacearum in Malaysia, p. 249. In Proceedings of the Fourth International Congress of Plant Pathology. Melbourne, Australia (Abstr.). ABDULLAH, H. 1988. Biology and Survival of Pseudomonas solanacearum in Malaysia. Ph.D. Thesis, 211 p. Univ. Agriculture Malaysia.

HAYWARD, A.C. 1960. A Method for Characterizing Pseudomonas solanacearum. Nature, London 186: 405-406. HA WARD, A.C. 1964. Characteristics of Pseudomonas solanacearum. J. Appl. Bacteriol. 27: 265-277. HE,

L.Y., L. SEQUIERA and A. KELMAN. 1983. Charac-

teristics of Strains of Pseudomonas solanacearum from China. Plant Dis. 67: 1357-1361. JOHNSTON, A. 1960. A Host-list of Plant Diseases in Malaya. Mycological paper no. 77. C.M.I., Kew, England.

BANCROFT, C.K. 1910. A Bacterial Disease of Potato and Tomato. Federated Malay States Agricultural Bull. 9: 478-480.

KELMAN, A. 1954. The Relationship of Pathogenicity of Pseudomonas solanacearum to Colony Appearance in a Tetrazolium Medium. Phytopathology 44: 693-695.

BUDDENHAGEN, I.W. and A. KELMAN. 1964. Biological

KING, E.O., W.K. WARD and D.E. RANEY. 1954. Two

and Physiological Aspects of Bacterial Wilt Caused by Pseudomonas solanacearum. Ann. Rev. Phytopath. 2: 203-230. BURNETT, F. 1949: Report on Agriculture in Malaya for 1947. CUPPELS, D., R.S. HANSON and A. KELMAN. 1978. Iso-

lation and Characterization of a Bacteriocin Produced by Pseudomonas solanacearum J. Gen. Microbiol 109: 295-303.

Simple Media for t h e Demonstration of Pyocyanin and Fluorescein./ Lab Clin. Med. 44: 301-307. NISHIYAMA, K., N.H. ACHMAD, S. WIRTOMO and

T.

TAMAGUCHI. 1980. Causal Agent of Cassava Bacterial Wilt. Control Center Res. Institute of Agric. Bogor Bull. No. 59. SINGH, K.G. 1973. Check-list of Hosts and Diseases in Peninsular Malaysia, Min. of Agriculture and Fisheries, Malaysia Bull. No 132.

GRAHAM, K.M., H. TAN, K.Y. CHONG, T.C. YAP, S.

VYTHIUNGAM. 1977. Breeding Tomatoes for Lowlands of Malaysia. Malaysian Appl. Biol. 1: 134.

12

THOMPSON, A. and A. JOHNSTON. 1953. A Host-list of

Plant Diseases in Malaya. Mycological paper No. 52. C.M.I., Kew, England.

PERTANIKAVOL. 15 NO. 1, 1992

Pertanika 15(1), 13-19(1992)

Kesan Sela Masa Pengawanan Semula dan Umur Sewaktu Pengasingan daripada Ibu ke atas Prestasi Reproduktif dan Tumbesaran Arnab New Zealand White CHONG, S. F., HANAFI BIN HARUN dan A. M. MAJID Jabatan Genetik, Fakulti Sains Hayat Universiti Kebangsaan Malaysia 43600 Bangi, Selangor Darul Ehsan, Malaysia Key Words: Remating interval, weaning age, rabbit performance. ABSTRAK Sebanyak 16 ekor arnab betina yang telah mencapai kematangan seks dan progeni mereka telah digunakan untuk mengkaji kesan sela masa pengawanan semula dan pengasingan daripada ibu ke atas dri-dri reproduktif dan tumbesaran arnab baka New Zealand White. Pengawanan semula dilakukan pada minggu 1, 2, 3 and 4 selepas kelahiran, manakala pengasingan daripada ibu dibuat ketika arnab berumur 3, 4, 5 dan 6 minggu. Sela masa pengawanan 'semula tidak mempunyai kesan yang signifikan ke atas kematian anak arnab dari waktu lahir hingga umur 3 minggu dan dari umur 3 hingga 12 minggu. Sela masa pengawanan semula juga tidak mempunyai kesan yang signifikan terhadap saiz kelahiran tetapi ia mempengaruhi (p

Fig. 2:

Temperature optimun of cellulase activities. Point represents an average of duplicates.

Effect of pH on Cellulase Activity

towards Filter Paper

Basically, fungal cellulases are stable from pH 3.0 to 8.0 at 30°C, active at pH 3.7 to 7.0 (Tong & Cole 1982). The optimal pH observed for cellulases produced by Aspergillus sp. occurred at pH 4.5 (Fig. 3). At a lower pH of 3.0, there was a sharp decrease in the cellulase activity to about 35% of the maximum. At a pH higher than 4.5, a gradual decrease in the activity was observed with only 9% of the maximum activity remaining at pH 8.0 and 9.0. In some fungal species, such as Trichoderma koningii (Wood 1969) and Aspergillus sp. (Khatijah et al 1983) double pH optimum has been reported.

The amount of extracellular cellulolytic enzymes in the crude filtrates was subsequently determined. It is interesting to note that there is no correlation between the amount of mycelial growth and the amount of cellulase enzymes produced. Xylan and CMC which did not promote much mycelial growth turned out to be good carbon sources in enhancing the production of cellulase enzymes. Other carbon sources which yielded good cellulase activity included cellobiose and cellulose powder. This supported the findings of Reese and Mandels (1971) that cellulase enzymes TABLE 1. Effect of different carbon sources on the mycelial growth. Carbon source

Effect of Different Carbon Sources on the Mycelial Growth and Production of Cellulase Enzymes

Xylan

To test the effect of different types of carbohydrate as the sole carbon source on mycelial growth and on the production of extracellur cellulolytic enzymes, culture was inoculated into Fergus (1969) medium and incubated for 15 days at 37°C. The results are shown in Table 1 and Fig. 4. There were some difficulties experienced in measuring the amount of mycelial dry weight in some of the substrates used. This was due to the fact that some of the mycelia became adsorbed onto the substrate thereby making its separation difficult Therefore, a visual obervation of the mycelial growth in the liquid medium was adopted. Cotton, cellobiose, filter paper and particularly cellulose powder were found to be good sources for the mycelial growth, whereas xylan, citric acid and mannitol were less suitable for mycelial growth.

Citric acid

48

Mannitol CMC Yeast Cotton Cellobiose Filter paper Cellulose powder

PERTANIKA VOL. 15 NO. 1, 1992

Arbitrary scale

EFFECT OF CARBON AND NITROGEN SOURCES ON THE GROWTH & PRODUCTION OF CELLULASE ENZYMES

production of cellulase enzymes in Fergus (1969) medium with filter paper as the carbon source was determined. As shown in Fig. 5, sodium nitrate, sodium nitrite and ammonium nitrate were found to be the best nitrogen sources for mycelial growth. However, ferrum ammonium sulphate and ammonium molybdate were not suitable for the growth of Aspergillus sp. Surprisingly, peptone did not support good mycelial growth either. 0

1

2 3 Activity (Unit/ml)

Fig. 4: Effect of different carbon sources on the production of cellulase activities. All values are average of duplicates.

were produced when cellulose was present as the carbon source in submerged cultures of fingal mycelia. Other carbon sources which are easily assimilated and have been shown to promote fungal growth may not induce the production of cellulolytic enzymes (Mandels and Reese 1965; Noviella, 1966). In addition, Rautela and King (1965), as well as Fan et al (1982) have shown that different forms of cellulose vary from one another in their growth-supporting, enzyme-inducing and reacting capabilities. This may be due to the difference in the microstructures among the cellulose substrates (Wang 1982). The cellulase activity observed may not be due to the actual amount of enzymes secreted in the cultures. This is due to the fact that cellulases have an affinity for certain cellulose sources. The cellulase enzymes would then be adsorbed onto the cellulose substrate thereby reducing the amount of detectable cellulase enzymes in the culture filtrate (Halliwell 1961). The percentage of this adsorbed cellulolytic activity has been estimated to vary from 0 to 40% depending on the species of the fungus. Greaves (1971) reported that by successively washing and gently agitating the mycelium and cellulose several times, the cellulase enzymes can be released from cellulose. Similarly, certain cellulose substrates, for example, the non-absorbent cotton wool, have surfaces that are not suitable for the absorption of cellulolytic enzymes leaving a higher amount of enzymes detectable in the filtrates. Effect of Nine Different Nitrogen Sources on the Mycelial Growth and Production of Cellulase Enzymes

The effect of nine different nitrogenous compounds on the mycelial growth of Aspergillus sp and

NaN03 N3N02

0.4

0.6

0.8

1

Hycelial weight (g)

Fig. 5: Effect of different nitrogen sources on the mycelial growth. All values are average of duplicates.

1 X

M

f

1 1

{ f

!

r

I

11 1

Fig. 6: Effect of different nitrogen sources on the production of cellulase activities. All values are average of duplicates.

With regard to the cellulase activity, again there was no correlation between the mycelial growth and cellulase production (Fig. 6). Ferrum ammonium sulphate heptahydrate and glycine stimulated maximum production of the cellulase enzymes followed by ammonium sulphate, urea, sodium nitrite and ammonium nitrate. Although ammonium nitrate supported the best mycelial growth, it did not stimulate as high a level of

PERTANIKAVOL. 15 NO. 1, 1992

CHOW-CHIN TONG AND K RAJENDRA

cellulase production as that achieved by ferrum ammonium sulphate heptahydrate and glycine. This may be explained by the fact that ammonium ions were absorbed by the mycelium which resulted in a lowering of the p H that subsequently inhibited the production of cellulase enzymes (Wang 1982).

NELSON, N. 1944. A Photometric Adaptation for the Somogyi Method for the Determination of Glucose. J. Biol. Chem. 153: 375-380. NOVIELLA, C. 1966. Pectolysis and Cellulolytic Activities of Sclerotium rofsii. Ann, Fac. Sri. Agron. Univ. Studi. Napoli Portia 30: 461-474. RAUTELA, C. 8C K.W. KING. 1965. Significance of the

Crystal Structure of Cellulose. Arch. Biochem. Biophys. 123(3): 599-601.

REFERENCES FAN, L.T., Y.H. LEE & M.M. GHARPURAY.

1982. The

Nature of Lignocellulosics and Their Pre-treatment for Enzymic Hydrolysis. Adv. Biochem. Eng. 23: 157-187. FERGUS, C. L. 1969. The Cellulolytic Activity of Thermophilic Fungi and Actinimycetes, Mycologia 61: 120-129. GREAVES, H. 1971. Effect of Substrate Availability on Cellulolytic Enzyme Production by Selected Wood-rotting Microorganisms. Aust. J. Biol. Sci. 24(6): 1169-1182. HALLIWELL, G. 1961. The Action of Cellulolytic Enzymes from Myrothedum verrucaria. Biochem. J. 79: 185-192.

REESE, E.T. and M. MANDELS 1971. Degradation of

Cellulose and its Derivatives. New York: John Wiley. SHEPHERD, M.G., A.LJ.

COLE & C.C. TONG. 1988.

Cellulases of Thermoascus aurantiacus. Methods in Enzymology 160: 301-307. SOMOGYI, M. 1952. Notes on Sugar Determination./ BioL Chem. 195: 19-23. TANAKA, M,, T. MORITA, M. TANIGUCHI, R. MATSUNO &

T. KAMIKUBO, 1 9 8 0 . / Ferment Tech. 58: 517. TONG, C.C, A.LJ. COLE 8C M.G. SHEPHERD. 1980. Pu-

rification and Properties of the Cellulases from the T h e r m o p h i l i c Fungus, Thermoascus aurantiacus. Biochem. J. 191: 83-94. TONG, C.C. 8c A.LJ. COLE 1982. Cellulase Production

HUDSON, JA., H.W. MORGAN & R.M. DANIEL. 1990. A

Survey of Cellulolytic Anaerobic Thermophiles from Hot Springs. Syst. Appl. Microbial 13: 72-76. KHATIZAH, M., M. I. YAZIZ 8C C.C. TONG. 1983. Deg-

by the Thermophilic Fungus, aurantiacus. Pertanika 5: 255-262.

Thermoascus

WANG, C.W. 1982. Cellulolytic Enzymes of Volvariella volvacea. p. 167-185. Hong-Kong : The Chinese University Press.

radation of Cellulose by Aspergillus sp., Trichoderma koningii and Myriococcum sp. Pertanika WOOD, T.M. 1969. Cellulolytic Enzymes System of 6: 8-16. Trichoderma koningii. Separation of Components MANDELS, M. 8C R.T. REESE. 1965. Inhibition of a Attacking Native Cotton. Biochem. J. 109: 217-277. Cellulase. Ann. Rev. Phytopath 3: 85-102. (Received 29 November 1991)

50

PERTANIKA VOL. 15 NO. 1, 1992

Pertanika 15(1), 51-54 (1992)

Solvent Extraction of Cd(II) using n-Butylamine-C10~-MIBK and n-Butylamine-Oxine-MIBK W.T. TAN AND S.K. WONG Chemistry Department, Faculty of Science and Environmental Studies Universiti Pertanian Malaysia 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia. Key words: absorbance, % extraction, pH, concentration, cadmium, n-butylamine, oxine, perchlorate, MIBK. ABSTRAK Pengekstrak n-butilamina-perchlorate-MIBK dan n-butilamina-oxine-MIBK didapati lebih berkesan untuk mengekstrakkan ion Cd(II) dalam keadaan alkali kuat (pH > 10). Pengekstrakan ini adalah fungsi kepekatan oksin, perklorat, n-butilamina dan pH. Isyarat nyalaan spektrometrik (AAS) yang lebih tinggi dapat ditunjukkan dengan menggunakan pengekstrak tersebut Peningkatan isyarat ini dan % pengekstrakan dapat diterangkan oleh pembentukan aduk amin sinergi.

ABSTRACT n-Butylamine-perchlorate-MIBK and n-butylamine-oxine-MIBKextractants were found to be effective for the extracti of Cd(II) ions under strongly alkaline conditions (pH>10). This extraction is a function of the concentration of oxine, perchlorate, n-butylamine and pH. The flame AAS signal for Cd(II) was considerably enhanced by using the above extractants. The signal enhancement and the increase in % extraction of Cd(II) can be attributed to the formation of synergistic amine adducts. INTRODUCTION In our previous report (Tan and Wong 1990), we described the advantages of using the primary short chain amine, n-butylamine, as a mixed solvent or complexing agent in the ammonium pyrollidine-dithiocarbamate (APDTC) -methyl isobutyl ketone (MIBK) extraction system. Higher extraction efficiency, higher signal output and greater stability of metal complexes were observed for the extraction of Cd(II) and Zn(II) ions from the n-butylamine-APDTC-MIBK system using flame atomic absorption spectrometry for metal analysis. As a continuation of our previous work, we shall report two other species which indicate same effects as APDTC in the presence of nbutylamine-MIBK but with increased signal output. These are perchlorate and oxine (8hydroxyquinoline). As in the case of APDTC and iodide, the use of oxine as a metal extractant has been widely reported (Marcus and Kertes 1969). However, there have been no reports on work done using

n-butylamine in the oxine-MBIK and perchlorateMIBK extraction systems. MATERIALS AND METHODS Reagent grade MIBK, nitric acid, sodium hydroxide, acetic acid, oxine, and perchloric acid were used without futher purification; n-butylamine was redistilled before use. An oxine solution (1% W/V) was prepared by dissolving 1 g of oxine in the minimum amount of acetic acid and adjusting to 100 ml using deionised distilled water. Fresh solutions were prepared as required. A stock solution of 1000 u.g/mL Cd was prepared by dissolving the pure metal in a minimum amount of nitric acid. The solution was standardized using EDTA. Solvent Extraction Solutions (40-50mL) containing known concentrations of metal ions and oxine or metal ions and perchlorate with adjusted pH were transferred to

W.T. TAN AND S.K. WONG

a separatory funnel containing 10 ml of the organic solvent(s). The pH of the solutions was adjusted using NaOH or HNO r The ionic strength of solutions was adjusted to 0.1 M with KNO^ when necessary. The solutions were shaken well for 1 minute and the phases were allowed to equilibrate for 5 minutes before separation was carried out. A flame atomic absorption spectrophotometer (Model IL 651) was used for metal determination. Absorbances obtained were corrected by using the reagent blanks. The percentage of extraction was calculated as follows:

100 D E= D + V/V where Vg and Vo refer to the volume of aqueous and organic phase respectively. D is estimated by using the ratio of the absorbances in the organic solvent to that of the aqueous phase. (Brooks et al 1989).

Effect of Perchlorate Concentration

The results in Table 2 indicate the requirement of a counter ion such as perchlorate for complete extraction of Cd(II) by the n-butylamine - MIBK system. The absorbance and hence the % extraction increases sharply from a low concentration of 1x10* M C10~ to near 100% extraction at 4xlO2M C10~ with a limiting absorbance value of 0.54. The optimum perchlorate concentration for the extraction of 1 |Ug/mL Cd is 0.04M at a fixed amine concentration of 7%. TABLE 2. Effect of varying (C10~) on the extraction of 1 |ig/ ml Cd(II) using 7% n-butylamine- C10~MIBK extractants at pH 11.4. (C10") (mM)

Absorbance

0.01 0.10 1.0 5.0 10.0 40.0 70.0 90.0

0.01 0.04 0.16 0.44 0.49 0.54 0.54 0.54

2 9 32 78 89 98 98 99

RESULTS Effect of n-butylamine Concentration n-Butylamine-C 10 ~-M IBK

pH Effect Results in Table 1 show that the extraction of Cd(II) ion by n-butylamine-C104-MIBK is strongly favoured under strongly alkaline conditions corresponding to the pl^ of n-butylamine (11.0). Rather poor extraction was observed under slightly alkaline and acidic conditions. Hence it is recommended that the natural pH of n-butylamine be used for chemical analysis. This pH condition was also used in subsequent studies. TABLE 1 Effect of pH on the extraction of 1 |4.g/mL Cd(II) using 7% n-butylamine- 40mM C10"4-MIBK pH

Absorbance

1.8 4.8

0 0.10 0.12 0.42 0.53 0.59

7.0 9.5 10.0 11.4

52

%E 0 9 10

70 90 100

The data in Fig. 1 show that metal extraction is dependent on the n-butylamine concentration. In the presence of 0.04M C10~4, 3% amine is sufficient to produce optimum signal output. n-Butylamine-Oxine-MIBK Extractants

pH Effect Results in Table 3 show that 1 jLlg/mL Cd(II) ion appears to be poorly extracted by the 0.01% oxine-MIBK extraction system. Although the efficiency of extraction increased with pH, the optimum pH of 11.4 only corresponds to 22% extraction of Cd(II) ion and 0.18 absorbance. However, in the presence of 0.5% n-butylamine, the extraction of Cd(II) ion increases to 80% which corresponds to an absorbance of 0.51 at pH ^ 10. It is therefore evident that the extraction of Cd(II) ion by oxine-MIBK is considerably enhanced in the presence of n-butylamine. Effect of n-butylamine Concentration

The effect of varying the initial concentrations of n-butylamine at a fixed concentration of 0.01% oxine and natural pH of 11.4 was studied. The

PERTANIKAVOL. 15 NO. 1, 1992

SOLVENT EXTRACTION OF CD (II) USING N-BUTYLAMINE-C10, -MIBK & N-BUTYIAMINE-OXINE-MIBK

TABLE 4 Effect of varying n-butylamine concentration on extraction of 1 |Ug/mL Cd(ll) using n-butylamine - 0.01% Oxine-MIBK extractants. n-butylamine 0.01 0.05 0.10 0.50 1.0 5.0

7.00

Absorbance

0.1% oxine since precipitation occurs. Calibration Curve

The data in Fig. 3 demonstrate the usefulness of a preconcentration step using n-butylamine-C104MIBK as solvent extractants for Cd(II) compared with the direct nebulization of aqueous samples of Cd(II) ions in flame atomic absorption spectrometry. However, linearity of the calibration curve was observed below 0.4 ppm for the case with solvent extraction. As in the case of the previous study on n-butylamine-APDTC-MIBK, the present studies of C10~4-MIBK and Oxine-MIBK-in

PERTANIKA VOL. 15 NO. 1, 1992

W.T. TAN AND S.K. WONG

plex species. The presence of n-butylamine would stabilize the CdOx complex at pH > 9. Under strongly alkaline conditions, the unprotonated amine predominates and could encourage the formation of a more stable amine adduct probably via the following equilibria: M + L + Am ;==± ML + Am ^ M + L + Am ^

ML (Am)

M(Am) + L ^ M ( A m ) L

L = Ox or C10" Am = unprotonated n-butylamine

0.8 5.

1.6

1.2

[CADMIUM]

( P P M

,

Calibration graph ofCd(II): • aqueous solution (before extraction) nonaqueous solution (after extraction with butylamine - 0.04M C1(J4 - MIBK)

ti-

the presence of n-butylamine produce similar calibration curves for the extraction of Cd(II). Both systems produce readable signals with a considerable degree of linearity at a range of 0.01 to 0.3 |lg/mL Cd(II). Under similar operating conditions, the Cd(II) signal was barely detectable via direct nebulization of aqueous samples containing less than 0.05 ug/mL Cd(II). DISCUSSION The explanation given previuosly (Tan and Wong 1990) for the enhanced Cd flame spectrophotometric signal using APDTC-MIBK in the presence of n-butylamine can be extended to the present studies. Oxine (8-hydroxyquinoline) has pKa values of 5.0 and 9.66 and its chemical equilibria are as given below :

The amine adduct, Cd(Ox) 2 (Am) or Cd(C104)2 (Am) thus formed would be easily extracted by the MIBK. This effect helps to explain the fairly high flame AAS signal observed for the Cd(II) ion and hence its extractability observed under strongly alkaline conditions rather than the alkaline condition and acidic conditions (Tables 1 and 3). The present and previous studies (Tan and Wong 1990) illustrate the advantage of adding nbutylamine to commonly used extraction systems such as oxine-MIBK and APDTC-MIBK especially for the extraction of Cd(II). The presence of nbutylamine in these extraction systems enhances the stability and the extractibility of the metal ion complex. It allows for the chemical analysis of Cd(ll) under strongly alkaline conditions which would otherwise prove impossible due to hydrolysis. ACKNOWLEDGEMENTS This work was supported by a Universiti Pertanian Malaysia Research Grant. The authors wish to thank C.B.Tan for technical assistance. REFERENCES BROOKS, R.R.,

M. HOASHI, S.M. WILSON and

R.Q.

1989. Extraction into MIBK of Metal Complexes with Ammonium Pyrrolidine Dithiocarbamate Formed in Strongly Acidic Media. Analytica Chimica Ada 217: 165 - 174. CHANG.

MARCUS, Y. and A.S. KERTKS. 1969. Ion Exchange and

Solvent Extraction of Metal Complexes, p. 515. UK:

J.W.Arrowsmith Ltd. The phenolate group and the unprotonated N group would be responsible for the chelation of'Cd(II) ion forming a five-membered ring chelate with the Cd ion to form the CdOx com-

TAN, W.T. and S.K.WONG. 1990. Enhancement of

Cd(II) and Zn(II) Flame Spectrometric Signals by using APDTC-nButylamine-MIBK Extractants. Pertanika 13(1): 95-99. (Received 10 July, 1991)

54

PERTANIKA VOL. 15 NO. 1, 1992

Pertanika 15(1), 55-59 (1992)

Calculation of the Effective Stopping Power of Ions Generated by Neutrons in Tissue Constituents ELIAS SAION Department of Physics Faculty of Science and Environmental Studies Universiti Pertanian Malaysia 43400 UPM Serdang, Selangor Darul Ehsan, Malaysia D.E. WATT Director and Safety Advisor Environmental Health and Safety Services University of St. Andrews St. Andrews, Fife, KY16 9TS, United Kingdom Key words: stopping powers, heavy charged particles, Ziegler's universal screening length. ABSTRAK Kertas ini melaporkan pengiraan kuasa penghenti zarah-zarah bercas berat yang tersentak oleh neutron dalam empatunsur kandungan tisu bagi tenaga zarah daripada 0.1 heV hingga 1.0 MeV. Pada tenaga rendah kurang daripada 30 keV/amu yang mana proses penghenti nukleus lebih berperanan daripada proses penghenti elektron, nilai kuasa penghenti berkesan lebih tinggi daripada nilai pendekatan perlambatan selanjarnya (PPS). Sisihan di antara kedua-dua nilai ini bergantung kepada jisim sasaran danjuga tenaga sertajisim zarah. ABSTRACT This paper reports the calculation of stopping powers of heavy charged particles generated by neutrons in four-element tissue constituents for particle energies from 0.1 keV to 1.0 MeV. At low projectile energies of less than 30 keV/amu where the nuclear stopping phenomenon is more dominant than the electronic stopping phenomenon, the effective stopping power values are higher than the continuous slowing-down approximation (CSDA) values, from which the deviation is dependent upon the target mass and the energy and mass of the projectiles.

INTRODUCTION Knowledge of stopping powers and projected ranges of low-energy secondary charged particles is very important for accurate prediction of energy deposition by intermediate energy neutrons in tissue (Al-Affan et al 1984). Intermediate energy neutrons slow down in tissue matter to generate short-range heavy charged particle recoils which have ranges less than the cellular dimension of most mammalian cells. The radiation quality of these neutrons, however, is not fully defined for radiation protection purposes although charged particles generated by these neutrons are known to be capable of producing biological effects (Jung and Zimmer 1966; Al-Kazwini et al 1988). Some have reported an increase in the radiobiological

effectiveness (RBE) of intermediate energy neutrons of energies less than 100 keV (Key 1971; Mill 1986). The penetration of charged particles through matter is a complex phenomenon which can be measured in terms of stopping power and projected ranges. In the region where projectile velocities, v, are greater than the electronic orbital velocity, v0 (= 27Ce2/ h = 25 keV/amu) the path length of ions in matter is a straight line. However, in the low energy region, i.e. v < v0, the pathlength is no longer a straight line but a complex form (Cook et al 1953) due to various interaction processes such as elastic collision with the target atom, inelastic collision with electrons, charge exchange and chemical binding effects. In general, there is competition between the loss of

PERTANIKA VOL. 15 NO. 1, 1992

55

ELIAS SAION 8c D.E. WATT

energy of charged particles to electrons and to recoil particles (Lindhard and Scharff 1961). The particles undergo successive small angle scattering known as multiple scattering which may produce recoils deflected in the direction inside the medium, increasing the stopping cross section per atom. The scattering angle becomes larger as the particle energy decreases. The particles travel in a zig-zag path which partly explains the difference between the projected range and path length of ions at low energies. The difference is very significant at energies where quasi-elastic scattering becomes dominant (Watt 1972). The projected range is an average thickness of material traversed by a number of particles directed perpendicular to the material surface. The effective stopping power is the energy loss per unit length along this projected range. For low energy ions the effective stopping power value is greater than the continuous slowing down approximation (CSDA) value because of the significant importance of elastic scattering collisions with nuclei of the target materials and range of straggling. In the region where nuclear stopping becomes more dominant than electronic stopping, and experimental stopping-power data is lacking, the stopping process of charged particles in matter is not fully understood and here, an attempt is made to include the effective nuclear stopping in the general stopping power formula. However, for ions at high energies, the effective stopping power will be the same as the CSDA stopping power because the loss of energy there is mainly due to collisions with atomic electrons. MATERIALS AND METHODS Calculation of Effective Stopping Power

is given as f(x) =0.021 [In ( m . / m ) ] 2 0.1434 [In (m./m)] +0.3642

Ziegler et al (1985) introduced a new universal screening length, az •= 0.8854a0/(Z.023 + Z.023), where Z. and Z are the atomic number of recoil ion type i and target atom type j respectively. The corresponding screening function produces a tight grouping with a standard deviation of 18%, better than the one derived from a well-known Lindhard's screening length, a( = 0.8854a 0 /(Z. 2/3 + Z 2/3 ) 1/2 , which has standard deviation of 40%. They found that their fitting universal screening function is in good agreement with the experimental data within 5% of the standard deviation. The reduced energy, £ , for ions of energy E (keV), in terms of Ziegler's screening length is given by l; E 23 , 7 0.2S,

8.462xlO" 15 Z i Z i m 1 (4) where Sn and Se are reduced nuclear and electronic stopping power respectively. The reduced nuclear stopping power is given by Ziegler in equations (5) and (6) for low and high energy ions respectively. Sn(e 2 ) In (1+ 1.1383 zz)

/

\

ln

type j respectively, y2 = 4 ra m. / (m. + m.)2 with ra and m being masses of particle i and target j respectively. f(x) is the function which converts the effective stopping power of nuclear stopping and

(ez) for £

(1)

where E is the energy of the recoil particle, S.n (E) and 'S e (E) are the nuclear and electronic stopping powers of recoil particle type i in the atom

(5)

for £2 < 30 keV/amu c

,sefl

56

(3)

The total stopping power is the sum of the nuclear and electronic stopping powers expressed in terms of Ziegler's reduced energy by

The effective stopping power, §.** (E), can be determined according to Watt and Sutcliffe (1972) in a manner similar to that adopted by Al-Affan et al (1984).

j

(2)

*

> 30

keV/amu

The corresponding reduced electronic stopping power can be expressed accordingly in terms of Ziegler's screening length by 7.93 x lO"2 Z2/3 Z^ (m, + m j " 2

L

£

"(V)

(Note the occurrence of T°n replacing Z 1 / 3 in the

PERTANIKA VOL. 15 NO. 1, 1992

CALCUIATION OF THE EFFECTIVE STOPPING POWER OF IONS expression given by Watt and Sutcliffe (1972)). In the high energy region, however, the contribution from nuclear stopping is very small and can be neglected. Therefore, one needs to consider only the electronic stopping power and this can be calculated according to the Bethe formula or by Ziegler's code (1985). The latter has been adopted in this work. The effective stopping powers of heavy recoil ions (i.e. protons, C, N, O) in tissue constituents (i.e. hydrogen, carbon, nitrogen and oxygen) have been determined. Except for recoil protons, the CSDA nuclear stopping power for C, N and O ions was calculated according to equations (5), (6) and the first term of equation (4). The electronic stopping powers of energies of less than 30 keV/amu were calculated according to equation (7) and the second term of equation (4). The choice of energy cut off corresponds to the ion velocity of about 2.25 x 106 m s"1. The electronic stopping powers for ions in the high energy region were calculated according to Ziegler's code. Normalization was performed to connect smoothly the stopping power values below 30 keV/amu to Ziegler's stopping powers for every ion interacting on each target. For protons in four-element tissue constituents, the CSDA stopping powers were obtained by fitting data of the International Committee on Radiation Units and Measurements (ICRU) (Berger 1986), for energies from 1 keV to 20 MeV. The fitting results produced deviation of less than 1%. Consequently, the effective stopping power of ions in four-element tissue constituents was calculated according to equation (1).

cada

A a +

I0"1

PROJECTILE ENERGY (keV) Fig. 1: The calculated effective and CSDA values of stopping powers for heavy ions (proton, C, N and O) in hydrogen CARBON

The effective and CSDA stopping power values calculated for recoil ions of energy from 0.1 keV to 1 MeV in hydrogen, carbon, nitrogen and oxygen are shown in Figs. 1, 2, 3 and 4 respectively. The effective and CSDA stopping power values are indicated by Seff (solid lines) and Scsda (broken lines) respectively. For nitrogen and oxygen ions, the values have been multiplied by factors of 10 and 100 respectively for graphic purposes only. The results are compared with published experimental data as indicated on each figure. The theoretical calculations of Oldenburg and Booz (1972) are also shown. The effective stopping power values, as expected, are higher than the CSDA stopping power values. As we can see from the figures, the deviation at the low energy region for a given recoil increases with the target mass and decreases as

n^-

x 0.01

N x 0. 1

" • • ' i

RESULTS AND DISCUSSION

Reynolds et o I . (1953) Falcuda (19B2) (1953) Oldenburg end Booz (1972)

en LU

-i—^7 „

10*

i

i

i

_ _ _' _ ^

i - t -

" H

- *

»-r^

c

;

r

o d

o _—

10 s

— **"

>-

o O

° o

10*

10°

A m o o o x K • M +

c

C

^eff iet ~a~I. ~(1953) ^csda Reynolds P h i l l i p s (19531 Vljngeerden I Duckworth 11962) So, Arkhlpov and Gott 11969) Sn+Se, Arkhlpov and Gott (1969) Moorhoad U965) Ormrod and Duckworth (1963) Hogborg (1971) FaBtrup ot a 1. 11 P66T Oldenburg Bnd Booz (1972)

101

10!

10 3

PROJECTILE ENERGY (keV) Fig, 2: The calculated effective and CSDA values of stopping powers for heavy ions (proton, C, N and O) in carbon

the projectile energy increases. Also, the deviation for a given target decreases with the mass of the projectile. The deviation is largest for protons

PERTANIKA VOL. 15 NO. 1, 1992

57

ELIAS SAION & D.E. WATT

NITROGEN x 0.01

-_ L - ' -

t.

I01

x 0. 1

£ o en

s

a: Q_ L3

5

in5

Seff S - " cSda Reynolds ot a I. 11953) Phi LLlps (1953) Fakuda (1980) Fakuda (1961) Oldenburg end Rooz (1972)

a. o

10 3 10"'

10°

10'

10 1

10 3

PROJECTILE ENERGY (keV) Fig. 3: The calculated effective and CSDA values of stopping powers for heavy ions (proton, C, N and O) in nitrogen 10

_

OXYGEN

D

x 0.01

:o8

component. The experimental stopping power data for recoil ions of low energies are generally lower than the present calculations due to the fact that the measurements were performed for electronic stopping powers. In one case when the calculated nuclear components were added to the experimental data of Arkhipov and Gott (1969), the results were consistent with the present calculation as shown in Fig. 2, for proton in carbon. Nevertheless, experimental data for most heavy ions in the low-energy region are scarce and are not sufficient to test the theoretical calculations. The present calculations consider the contribution from the effective nuclear stopping power which is otherwise not included in the experiments. Our results suggest the need to study the contribution of effective nuclear elastic scattering in the penetration of charged particles in hydrogen, carbon, nitrogen and oxygen, from which the effective stopping powers and projected ranges can be calculated. Accordingly, the effective stopping power and projected ranges in tissue and tissue equivalent (TE) materials can then be extracted in the recommended manner. Both quantities are important in the determination of microdose spectra and quality of intermediate energy neutrons in the simulated tissue volume of Tissue Equivalent Proportional Counter (TEPC). CONCLUSION

10

The effective stopping power is higher than the CSDA stopping power for low energy ions due to interaction of particles through nuclear elastic scattering. The deviation for a given particle increases with the target mass and decreases with an increase in the projectile energy. For a given target, it decreases with the mass of the particle. The results can be used to predict the quality of intermediate energy neutrons which could show the possibility that elastic nuclear collisions play a significant role in determining an increase in RBE for these neutrons.

s

a

to'

QJ D CL

|,o.

Seff A • a +

Reynolds et a I. (1953) Phllllpa (19531 Fakuda (1982) Oldenburg and Booz (1972)

REFERENCES 10"'

10°

101

10*

105

AL-AFFAN, I. A. M, P. COLAUTTI, G. TALPO and D. E. PROJECTILE ENERGY

(keV)

Fig. 4: The calculated effective and CSDA values of stopping powers for heavy ions (proton, C, N and O) in oxygen

in all the targets. In the high energy region, however, the deviation is negligible because the nuclear stopping power component is very small in comparison with the electronic stopping power 58

WATT. 1984. Calculated Microdose Spectra for Intermediate Energy Neutrons (1 to 100 KeV). Radial Prot. Dosi 5(3): 151-157. AL-KAZWINI, A. T., J. W. CANNINGHAM and D. E. WATT.

1988. Damage by Nuclear Elastic Scattering (NES) - Forty Years On. Int. J. Radial. BioL 53(4): 683-685.

PERTANIKA VOL. 15 NO. 1, 1992

CALCULATION OF THE EFFECTIVE STOPPING POWER OF IONS

E. P. and Yu. V. GOTT. 1969. Slowing Down of 0.5-30 keV Protons in Some Materials. Sov. Phys.JETP 29(4): 1146-1151.

ARKHIPOV,

M.J. 1986. Proton Stopping-Power and Range Tables. ICRU Stopping Power Committee.

BERGER,

COOK, C.J., E.JONES and T.JORGENSEN.

1953. RangeEnergy Relations of 10-250 keV Protons and Helium Ions in Various Gases. Phys. Rev. 91(6): 1417-1422.

A. 1980. Stopping Powers of a Tissueequivalent Gas for 40-200 keV Protons. Phys. Med. BioL 25(5): 877-886.

FAKUDA,

A. 1981. Stopping Powers of a Tissueequivalent Gas for 40-200 keV He+ and N+. Phys. Med. Biol. 26(4): 623-632.

FAKUDA,

FAKUDA, A.

1982. Stopping Powers of H2, O2, C2H4 for 40-200 keV Protons. Phys. Med. BioL 27(1): 73-79.

FASTRUP, B., P. HVELPLUND and C. A. SAUTTER. 1966.

Stopping Cross Section of Carbon of 0.1-9 MeV atoms with ZYOj|Yn> ...,Y O = Xn daripada keputusan (2). Umbaian U2

(Diterima 11 Mei, 1991)

s E J

[ {T=k}Xk J daripada lema 2.3

= E [ x k | T = k] Kb [T =

Y I

^E[x n+k |Y n) ...,Y o j

Martingale dalam Gerakan Brownan. Sains Malaysiana 17(4): 481-490.

= E[l| T=k }E[x k+n _ k |Y k ,...,Y o ]]

Bukti lema 2.3 dengan aruhan.

Y

B. V. 1968: The Theory of Probability. New York: Chelsea Pub. Co.

GNEDENKO,

n |T

= k]>E[x k |T =

Umbaian U3 Bukti lema 2.5 Pertimbangkan peristiwa

Bukti Lema 2.4 Pertimbangkan E X ^TAn =E

+E

E[x n I{ T=k }|Y k! ...,Y o ]

n-1

X

=E

= I E I XT Ti {I

I| T=k jE[x n |Y kl ...,Y 0 ]

yang I 72

{

1 jika peristiwa c berlaku 0 jika sebaliknya . PERTANIKA VOL. 15 NO. 1, 1992

KEGUNAAN SUB-MARTINGALE DALAM MASALAH STORAN

Umbaian U4 n-l

Di tunjukkan

= IE

k=0 n-l < E E k=0

=1 yang{Yn}n=0 § tertabur tak bersandar sepercaman normal dengan min \i dan varians a 2 = 2(i/X.

Pertimbangkan jujukan

X o =0, X n = Z |x k -E[x k |Y k _ 1> ...,Y o ]j

n

2

M (e) = e

ixl maka |

Katakan M( 0) ialah Fungsi Penjana Momen N 1i |i.o2 iI yang fungsi penjana momennya diberi oleh

Maka persamaan

f ialah Martingale

M

E fe eY "l = 1

« 0=E

(e) = i

XTAn

= E

^S {xk-E[xk|Yk.lf...,Yo]}

M0

+

l

Q202 =

2 TAn

\ /; v^ ;

\

46.22 1800 Fig. I(b)and l(c)

1600

/ \ A

w

f\v

*/

1400

1200

1000

800 cm"1

Comparison FT-IR spectra of lignin obtained using the KBr pellet cell. (A) - Rattan; (B) Kaput; (C) - Rubberwood; (D) - Selangan Batu and (E) - Belian.

80.08%T

63.18 1800 1600 1400 1200 1000 800 cm"' Fig. 2 (a) : FT-IR spectrum of Selangan Batu wood strip obtained using the MIR technique. PERTANIKA VOL. 15 NO. 1, 1992

79

S. LAU & RAML1 IBRAHIM

62.19 1800 Fig. 2 (b) and 2 (c) :

80

1600 1400 1200 1000 800 cm*1 Comparison FI-IR spectra ofivood strips obtained using the MIR technique. (A) -Rattan; (B) - Selangan Batu; (C) - Rubbenuood; (D) - Kapur and (E) - Belian. PERTANIKAVOL. 15 NO. 1, 1992

FT-IR SPECTROSCOPIC STUDIES ON LIGNIN FROM SOME TROPICAL WOODS AND RATTAN

nificant differences among the various wood samples. T h e first was the absorption band of wave n u m b e r between 1500 and 1515 cm 1 . Belian wood showed a relatively strong absorption at 1510 cm 1 while the wood showed a doublet at 1503 a n d 1512 cm*1. This absorption band is assigned to the aromatic skeletal vibration. This indicates that belian has almost a single type of aromatic ring while the other wood contains at least two types with a more or less similar concentration. T h e second absorption band is at the region between 1000 a n d 1030 c m 1 . Belian a n d selangan batu showed a single absorption band at 1013 c m 1 while kapur showed a wider band which was unresolved at 1007 cm 1 . Rubberwood showed distinctly two equally strong bands at 1013 a n d 1030 cm 1 , while rattan also showed the two similar bands, with the band at 1030 cm'1 being much stronger than that of 1013 cm 1 . T h e absorption band at 1030 c m 1 is assigned to the aromatic C-H in plane deformation of t h e guaiacyl type (5). This indicates that lignins in rubberwood a n d rattan contain a higher proportion of guaiacyl group than those from belian, selangan batu and kapur. CONCLUSION This study has shown that there are detectable differences in the FT-IR spectra of the extracted lignins as well as in the untreated wood strips of various types of wood. Further studies should be carried out to elucidate the specific differences between the lignins of various woods. Separation of lignin components will be necessary to characterise the exact nature of each type of lignin in plants. Further work is being done in our laboratory.

REFERENCES BiTA, J.G., F. ZADRAZIL, and G.C. GALLKTTI. 1989. FT-

IR Determination of Lignin Degradation in Wheat Straw by White Rot Fungus Stropharia rugosoannulata with Different Oxygen Concentrations. J. Agric. Food Chem. 37: 1382-1384. F.ux. O. and O. BEONHOFF. 1988. FT-IR Spectra of Milled Wood Lignins and Lignin Polymer Models (DHP's) with Enhanced Resolution Obtained by Deconvolution. /. Wood Chem. TechnoL 8(4): 505-523. HARHN, J. M. 1969. Methods of Attacking the Problems of Lignin Structure. In Recent Advances in Phytochemistry, vol.11, ed. M. K. Seikel and V. C. Runeckles, p 35-75, N. Y.: Appleton-CenturyCrofts. HERGERT, H . L. 1 9 7 L In LIGNINS e d . K. V. SARKANEN and

C. H.

LUDWIG, p 267-293 N. Y. : Wiley-

Interscience. HlGUCHI, T. 1981. Lignin Structure and Morphological Distribution in Plant Cell Wall. In Lignin Biodegradation: Microbiology, Chemistry and Potential Applications, vol.1,pi-19 ed. T. K. Kirk, T. Higuchi and H. M. Chang, Florida: CRC Press JANTAN, M. D. and M. K. TAM. 1987. Natural Dura-

bility of Malaysian Timbers, Timber Trade Leaflet No. 28, FRIM, K. L. MORTON, J. and H.E. SPARKKS. 1967. Tappi, 50: 363. RITTER, G.J., R. M. SEBORG and R. L. MITCHELL. 1932,

Ind. Eng. Chem., Anal. Ed., 4: 202. ROY, A.K., S.C. BAG and S.K. SEN. 1987. Studies on the Chemical Nature of Milled Wood Lignin of Jute Stick. Cellu. Chem. TechnoL 21: 343-348.

ACKNOWLEDGEMENTS The authors wish to thank IRPA for funding the project and Encik Mohd Afandi bin Mat Sani and Encik Sidem Nogep for assistance. S. LAU 8c RAMLI IBRAHIM Centre of Basis Science Studies Universiti Pertanian Malaysia, Kampus P.O. Box 396, 97008 Bintulu Sarawak Malaysia. .

Bintulu (Received. 2 September 1991)

PERTANIKAVOL. 15 NO. 1, 1992

81

Section III Social Sciences

Pertanika 15(1), 85-91 (1992)

January Effect on the Thinly Traded KLSE: Tests with Appropriate Refinements M.N. ANNUAR, M. ARIFF1 and M. SHAMSHER Department of Accounting and Finance, Faculty of Economics and Management, Universiti Pertanian Malaysia, 43400 UPM, Serdang, Selangor Darul Ehsan, Malaysia. Department of Finance and Banking, National University of Singapore, No. 10 Kent Ridge Crescent, Singapore 0511 Keywords: January effect, thinness of trading, regularity. ABSTRAK Sejak kebelakangan ini Kesanjanuari telah dirakamkan sebagai gelagat anomali di kebanyakan pasaran saham di negara-negara rnaju. Kesan January berlaku apabila pulangan sekuriti pada awal bulan Januari melebihi pulangan pada bulan-bulan yang lain. Keivujudan Kesanjanuari di Bursa Saham Kuala Lumpur (BSKL) telah ditemui oleh satu kajian awal. Kajian ini adalah lanjutan kajian tersebut dan mengambil kira beberapa penghalusan yang wajar seperti mengkaji pulangan 298 saham (bukan mengkaji pulangan a (as indeks), mengkaji hubung kait kesan saiz dengan Kesanjanuari, dan mengkaji kesan faktor ketipisan dagangan. Penemuan kajian ini menunjukan bahawa Kesanjanuari tetap wujud bagi pulangan saham di BSKI, dan pulangan purata bersih sebanyak 1.75 peratus diperolehi dalam bulan Januari. Walaupun demikian, pulangan purata bersih ini belum diselaraskan risikonya: oleh itu kecekapan penUaian Kesan Januari belum dapat dipastikan. Berbeza dengan hasil penemuan di negara-negara maju, kesan saiz tidak dapat dihubungkaitkan dengan Kesanjanuari. Kemungkinan faktor ketipisan dagangan sebagai penjelasan wujudnya Kesan Januari disokong oleh kajian ini. ABSTRACT Over recent years the January Effect has been documented as an anomaly on stock markets around the ivorUL The January regularity refers to the phenomenon that security returns in early January are higher than in any other period of the year. The presence of the January regularity on the Kuala Lumpur Stock Exchange (KI^SE) has been established. Hoxoever, this study investigates further the possible explanations, taking into consideration the returns on stocks rather than indices, control for thinness of trading and the association of the regularity with size effects. The evidence indicates that returns on 298 stocks traded in KSIJt do exhibit the January seasonality, and the average returns net of cost for January are L 75 per cent. However, these net average returns are not adjusted for risk, therefore it is difficult to confirm the valuational efficiency of the regularity. In contrast to the documented evidence in developed securities markets, the size effect cannot rationalise the regularity in the developing Kuala Lumpur Stock Exchange. The possibility of the thinness of trading as a [(id or that could partially explain the regularity is supported. INTRODUCTION In recent years, a number of anomalies in market price behaviour have been documented in professional and academic journals. One such regularity is the January effect on stock markets around the world. This regularity was observed in Copenhagen Stock Exchange as far back as the 1890s (Jennergren and Sorensen 1988) but was only given serious attention in the mid-1970s. The

January regularity refers to the phenomenon that security returns in early January are higher than in any other period of the year. It has also been documented that average returns on stocks of small firms in January are relatively higher than those of large firms (Keim 1983, Gultekin and Gultekin 1983). However, the regularity has been noted in returns of a variety of share characteristics such as size, yield and neglected firm

M.N. ANNUAR, M. ARIFF & M. SHAMSHER

(Keim 1983, 1985, Arbel and Strebel 1983, Arbel 1985). Like all anomalies, the fanuar) regularity is documented as evidence of market inefficiency (Jacobs and Levy 1988). If this regularity implies a regular and predictable pattern of price behaviour, should not then the market arbitrage away such regularity? Keane (1989) explains that the regularity might explain informational inefficiency, but for the market to be valuationally efficient, the regularity must be material and persistent for a reasonable period of time. The evidence of market anomalies suggests that the informational definition of market efficiency by Fama (1970), " the market is efficient if it correctly reflects all available information " cannot be completely true of the real markets. For investment purposes the markets might be valuationally efficient as there is no evidence that the seasonalities are exploitable. A number of plausible explanations have been offered for the January regularity which at best rationalise investors' behaviour but do not explain how the January returns might be reconciled with market rationality. Keim (1983) and Reinganum (1983) suggest that a large part of the differential risk adjusted returns occur in the first week of January, consequently it might be that investors sell securities at the end of the year to establish short-term losses for income tax purposes and then buy securities in early January. This explanation suggests that there is a downward pressure on stock prices in late November and December and a positive impact in early January. This explanation assumes that capital gains are taxed as income and capital losses are deductible from taxable income. However, Constantinides (1984) argues that an optimal policy for investors who want tax shields against profits is to realise losses as they occur and not wait for the end of the tax year. Portfolio managers usually sell low performing securities at the end of the year to avoid their appearance on the annual report. Similar securities are repurchased in the new year. Rogalski and Tinic (1986) suggest that the regularity might be partially explained as compensation for higher risk and provide evidence that volatility of security prices in January is significantly greater than in other months. Arbel (1985) suggests the regularity might be a compensation for bearing informational risk. The effect may be due to the reduction of uncertainty associated with the dissemination of information after the end of the fiscal year for small and

neglected firms. However, Chari et al. (1986) provide evidence that companies with non-December fiscal year experience returns that are seasonal at the turn of the fiscal year, as information risk is resolved rather than at calendar year end. Jacobs and Levy (1988) suggest that cashflow patterns at the end of the year may explain the January regularity. Annual bonuses, holiday gifts and pension contributions might be invested in the stock market in January. However, this regular and predictable returns could be arbitraged. In summary, the literature on the regularity provides no convincing evidence of economic significance of the regularity. For investment purposes, this is sufficient reason for investors to behave as if the market is valuationally efficient. Objective of Study

The presence of the January regularity on the KLSE has been established (Annuar and Shamsher 1987). However, this study intends to investigate further the possible explanations, taking into consideration important characteristics of a developing securities market like KLSE. This study refines the previous study in the following respects: (i) It investigates the regularity on returns of 298 stocks rather than indices. The use of indices results in underestimation of the returns as dividends are not included. (ii) It controls for the thinness of trading of stocks at KLSE. Thinness of trading is a peculiar characteristic of developing securities markets. Failure to adjust for thinness of trading biases the results in favour of the null hypothesis of non-regularity for a given level of significance. Thinness is controlled by analysing the returns of stocks which are continuously traded in the market over the period of study. Eighty-two stocks qualified for this requirement. (iii) After controlling for thinness of trading, the association of the regularity with size effect is investigated. (iv) The presence of the regularity is investigated on the returns of the total sample (298 stocks), the controlled sample (82 stocks) and noncontrolled traded sample (298 less 82 stocks). If the presence of the regularity is observed only in the total and the non-controlled sample, this would suggest that thinness of trading might explain the regularity. (v) The valuational efficiency of the regularity (if present) would be determined, considering

PERTANIKA VOL. 15 NO. 1, 1992

JANUARY EFFECT ON THE THINLYTRADED KLSE

round-trip transaction costs of about 2.7 per cent on the KLSE. Prior Research

There is a dearth of published evidence on the January regularity of returns at the KLSE. Annuar and Shamsher (1987) showed the presence of the Januar)' regularity at the KLSE. However, they used indices instead of individual securities. Wong et al (1989) used six KLSE sectorial indices from 1970 to 1985 and showed the presence of the regularity, expressed in terms of the Gregorian calendar month effect. Yong (1989) also used six sectorial indices at KLSE from 1970 to 1988 and concluded that there is no January regularity on the KLSE. In view of mixed evidence prevailing, this study intends to investigate the presence of the regularity considering the appropriate refinements discussed earlier. MATERIALS AND METHODS Monthly price relatives were calculated for the 298 listed stocks on KLSE for the period January 1975 to June 1989. The returns are calculated as follows: R

log

P. + D

where Pjt is the price of the stock i at time period t. P kl is the price of the stock i at time period t-L The average monthly returns were computed for all the months over the 15-year period of study. The difference between January returns and returns for February to December were determined. Any significant difference would imply the presence of the regularity. To ascertain the possible association of the regularity with activeness of trading, 82 stocks were identified from ten deciles, ranging from the most active trading (decile 10) to the least active (decile 1). This sorting was done for each year and ten deciles were reformed each succeeding year to 1989. This results in ten deciles of stocks with broadly similar frequency of trading within each decile, such that the returns calculated in a particular decile are free of thin-trading bias. Thinness of trading is measured by normalising the annual volume of trading by the average number of shares outstanding.

For each decile, the difference between January returns and returns for February to December was observed and the difference was tested for significance using normal test statistics. To ascertain the possible link between the regularity and size of firms, the 82 controlled stocks were categorised into small and large firms based on their market value of stocks at the end of each year. The presence of the anomaly was tested on the samples of large and small firms. The following hypotheses were tested: (1) The presence of the January anomaly on returns of 298 listed stocks at KLSE. The presence of the regularity will be evidenced by the significant positive difference between average January returns and average returns of February to December. (2) For the sample of controlled stocks, the presence of the regularity on returns of stocks with different activeness of trading was investigated. If there is no evidence of the regularity on the most and the least actively traded stocks, it would imply that thinness of trading might partially explain the regularity. (3) The presence of the regularity on returns of different sizes of firms. If the regularity is present only on returns of small firms, this would imply that size might partially explain the regularity. (4) The economic significance of the regularity is then evaluated. If the regularity is present, could investors devise a trading strategy to earn abnormal gains net of the round-trip costs of about 2.7 per cent. RESULTS Tables 1 (a) and (b) summarise the mean returns of each month for the 298 stocks, for the period 1975 to 1989. The average monthly percentage returns are positive for all the months. The January returns of 4.45 per cent are about twice as large as the average returns of February to December, and the difference is significant at 5 per cent level. This evidence is consistent with the findings of our earlier research (Annuar and Shamsher 1987), which implies that the presence of the regularity cannot be attributed to few outliers as the number of positive returns dominate the negative returns in January. It is also possible that large returns in January could be due to higher total risk on stock returns, as measured by variances of returns.

PERTANIKA VOL. 15 NO. 1, 1992

87

M.V \ \ \ (

TABLE l(a) Average monthly percentage returns 1 298

KR. M. ARIFK& M. SHAMSHER

listed

stocks on KLSK: 1975-1989 Month

Return

Variance

Jan nan

4.45 2.75 1.38 3.79 3.24 1.42 1.46 0.35 0.22 0.16 2.86

0.0023 0.0014 0.0017 0.0032 0.0025 0.0026 0.0012 0.0023 0.0034 0.0012 0.0061 0.0043

February

March April May

[une Jnlv

August September October November December

L.96

TABLE l(b) Average January percentage returns versus average percentage returns of February through December of 298 stocks: 1975-1989 January 4.45

February-December

I-si at

2.06

2.47 *

For the next most active decile (decile 9), the average returns for January are positive (5.1%) but not significantly different from average returns of February to December (t=l .46). However, the F-statistics (Fall) and (Fj) are significant at 5 per cent level implying that there is a significant difference in the average returns of each month. For the least actively traded stocks (decile 1), returns are positive for all months, but there is no evidence of significant difference in the returns of each month (FiU = 1.33 and F( = 1.39). The average January returns are not significantly different from the average returns of February to December (t=1.63). For the intermediate deciles (decile 4 and 5) there is evidence of significant difference between average January returns and average returns of February to December at 5 per cent level. This implies that, on all average, January regularity might be confined to stocks with moderate activeness. Overall, the absence of the regularity in eight out of the ten deciles (based on t-statistics and Ftests) suggests that thinness of trading might explain the regularity. January Effect on 216 Non-controlled Stocks

However, Table 1 (a) shows that for the sample studied, variances of returns cannot explain the higher January returns. The variance of returns for January is 0.23 per cent, which is lower than all months except February, March, July and October. Table 1 (a) also shows that the returns in October were on average significantly lower than any other month of the year for the 15-year period, consistent with the October effect, reported by Cadsby (1988).

The presence of the regularity is investigated on the total sample of 298 stocks less the 82 controlled traded stocks. The findings are summarised in Tables 3(a) and 3(b). The average monthly percentage returns for all the months except September are positive. The average returns for January are three times as large as the average returns of February to December (4.31% versus 1.4%), and the difference is significant at one per cent level (t=3.28). This further supports the notion that thinness of trading might explain the regularity.

January Effect and Frequency of Trading

January Effect and Sizes of Firms

Table 2 shows the average monthly percentage returns on portfolios of stocks with different frequency of trading. For the most actively traded decile (decile 10), the returns for each month are positive except for the month of October. Irrespective of including (F H) or excluding (F) the month of January, there is no significant difference in the returns of each month in this decile. The average returns in January (5.4%) the fourth largest after the returns in November, April and September, are not significantly different from the average returns for the months of February to December (t=1.61).

Tables 4 (a) and (b) show that for samples of both large and small firms, the average returns for January are significantly different from the average returns of February through December at 5 per cent level (t=2.19 and 2.60 respectively). This evidence suggests that size effect alone cannot explain the regularity, which is inconsistent with the documented evidence on developed securities markets. The pronounced January effect on returns of small firms documented in developed securities markets (Keim 1983), could be due to relative thinness of trading of smal1 finns' stocks.

^significant at 5 per cent level.

PERTANIKA VOL. 15 NO. 1, 1992

TABLE 2 Average monthly percentage returns on portfolio stocks with different levels of trading: 1975-1989 Month Sept

Ocl

Nov

Dec

Average (Feb-Dec)

0.765

5.887

-3.393

7.817

4.435

3.569

0.560

-1.00

-2.136

1.245

0.956

5.005

3.145

8.033

1.670

-0.192

-0.509

-0.958

4.951

0.020

4.260

-1.386

4.080

-1.269

3.048

3.318

2.261

0.934

1.584

0.036

3.233

4.431

0.469

2.181

-1.107

1.363

3.033

1.339

4.367

1.512

0.505

-0.448

3.696

0.887

3.876

2.672

3.347

-0.166

3.316

4.023

3.360

3.327

1.213

1.752 Decile 1 (I .east Active)

0.917

3.649

1 ',07

1.467

Apr

May

June

July

(Most Active) 5.425 Decile 10

2.221

3.66

7.789

3.991

2.765

3.283

Decile 9

5.124

2.482

4.505

3.905

2.678

0.443

PERT i,VOL

r-

Dedle 8

6.949

1.504

1.933

2.893

2.650

Decile 7

2.534

1.666

2.651

5.048

Decile 6

6.637

1 .\)^2

3.669

c (£ 35 KS

Decile 5

7.404

2.379

Decile 4

5.149

Decile 3 Decile 2

F

t-tcst

0.90

0.84

1.161

1.694

2.44*

2.50*

1.461

•n

4.866

2.548

1.65

1.40

1.56

:T OX

Mai

ANUARY i

Fob

|«m

-1.312

3.127

2.012

1.08

1.13

0.38

H

0.408

-0.818

4.821

1.928

1.51

1.14

1.83

H X

0.166

0.277

-0.223

3.728

1.536

1.21

1.05

1.98*

_j

1.382

3.152

-0.300

-0.132

5.366

1.797

1.71

1.55

2.22*

0.803

0.367

-0.331

-0.174

0.687

4.681

1.514

1.89*

1.76

1.24

2.329

-0.119

0.499

1.019

-1.388

1.498

3.753

1.790

1.76

1.54

0.94

0.838

1.186

1.209

1.137

0.923

4.826

1.619

2.007

1.33

1.39

1.63

70

* significant at 5 per cent level

5

M.N. ANNUAR, M. ARIFF & M. SHAMSHER

TABLE 3(a) Average monthly percentage returns of 210 stocks on KLSE: L97M989 Month

January February March April May June July

August September October November December

Return

Variance

4.31 2.88 0.69 3.64 0.40 1.41 1.35 0.24 -0.18 0.38 3.47 1.13

0.0010 0.0005 0.0010 0.0025 0.0032 0.0021 0.0008 0.0019 0.0022 0.0005 0.0067 0.0002

TABLE 3(b) Average January percentage returns versus average percentage returns of February through December of 216 stocks: 1975-1989 January 4.31

February-December

t-stati sties

1.40

3.28*

* Significant at 1 per cent level

DISCUSSION The presence of the January regularity was investigated and supported on returns of 298 stocks listed on the KLSE. The average January returns exceed the round-trip transaction costs of 2.7 per cent. However, the average January returns net of transaction cost are not adjusted for risk, thus the val national efficiency of the net return cannot be confirmed. The total risk of security returns in January was lower than those of most other months, implying that it cannot rationalise the regularity. The January effect was investigated on a sample of controlled stocks. There was no evidence of the regularity in both portfolios of most actively traded and least actively traded stocks. Only the moderately active portfolio of stocks exhibited the presence of the regularity. Since only two out of ten deciles exhibited the regularity, it supports the suggestion that thinness of trading might partially explain the regularity . Thus it is possible that the apparent January regularity documented in developing securities markets could be a proxy for the effect of thinness of trading. 90

TABLE 4(a) Mean average percentage returns for large and small firms: 1975-1989 Large Firms

Small Firms

April May June July August September October November December

5.169 0.333 2.118 5.539 3.413 2.826 2.073 0.273 1.989 -1.378 -0.346 2,689

4.157 1.935 2.475 3.303 1.346 1.932 1.257 -0.228 0.437 -0.063 -0.327 2.391

February to December

1.775

1.223

1.34 1.39

1.23 1.18

January February March

•i

TABLE 4(b) Average returns in January versus average returns of February through December: 1975-1989 Group

January February to December t-statistic

Large firms

5.169

1.775

2.19*

Small firms

4.157

1.223

2.60*

* Significant at 5 per cent level

Since Keim's (1983) study indicated that much of the year's size effect seemed to occur in January, a series of papers by Banz (1981), Keim (1985), Roll (1982), and Keim and Stambaugh (1984) showed that all of the small companies' effect in U.S. virtually occurs at the end of December and early January. They tend to rationalise the January regularity in terms of small firm effect. The sample in this study was divided into 'small' and 'large' firms based on the market value of the outstanding stocks at the beginning of each year. The presence of the regularity in both samples was investigated and supported. This implies that size factor alone cannot explain the January regularity. However, it is possible that the January regularity was reinforced by the small firm effect but certainly the size factor per se cannot explain the regularity on stocks traded in KLSE. The regularity is observed on the returns of the total sample, and the non-controlled sample but was not present in the controlled sample.

PERTANIKA VOL. 15 NO. 1, 1992

JANUARY EFFEC I ()N THE THINLYTRADED KLSE

This suggests that to mitigate the possibility of implausible findings, empirical studies on the January regularity in developing securities markets should control for the thinness trading. CONCLUSION The returns on stocks traded on the KLSE do exhibit the January seasonality, and the average returns n e t of cost for January are 1.75 per cent. However, as these net average returns are not adjusted for risk, it is difficult to confirm the valuational efficiency of the regularity. In contrast to the documented evidence in developed securities markets, the size effect cannot rationalise the regularity as both samples of large and small firms exihibited large and positive significant returns in January vis-a-vis the returns of February to December. The possibility of thinness of trading as a factor that could partially explain the regularity was investigated and supported. This suggests the necessity of controlling for thinness of trading in any empirical study on the January anomaly in developing securities markets. REFERENCES ANNUAR, M.N. and M. SNAMSHKR. 1987. The January

Effects on Stocks Traded on the Kuala Lumpur Stock Exchange: An Empirical Analysis. Hong Kong journal of Business Management 5: 37-52. ARBEL, A. and P. STREBEL. 1988.

Pay Attention to

Neglected Firms. The Journal of Port]olio Management 9: 37-42. ARBEL, A. 1985. Generic Stocks: An Old Product in a New Package. The journal of Portfolio Management 11: 4-13. BAN/, R.W. 1981. The Relationships between Returns and Market Value of Common Stocks./ Finan. Earns. 9: 3-18. BILDERSEE, J. and N. KAHN. 1987. A Preliminary Tost

of the Presence of Window Dressing, journal of Accounting, Auditing and Finance. 44: CADSBY, C.B. 1988. The CAPM and the Calendar Anomaly: A Systematic Treatment of Empirical Anomalies. University of Guelph, Discussion Paper.

CHARI, V., R. JOGANNATHEN

and A. OFER. 1986. Fiscal

Year End and the January Effect. Kellogg Working Paper #20, Northwestern University. CONSTANTINIDES, G.M. 1984. Optimal Stock Trading with Personal Taxes: Implication for Prices and the Abnormal January Returns. /. Finan. Earns. 13: 65-89. FAMA, E.F. 1970. Efficient Capital Markets: A Review of Theory and Empirical Evidence. /. Finan. 25: 383-417. GULTEKIN, M. N. and N. B. GULTEKIN. 1983. Stock

Market Seasonality: International Evidence. /. Finan. Earns. 12: 469-481. JACOBS, B. I. and

K. N.

LEVY.

1988.

Calendar

Anomalies: Abnormal Returns at Calendar Turning Points. / . Finan. Analy. 44: 28-39. JENNERGREN, L. and P. SORENSEN (1988), quoted in

Keane, S. M. Seasonal Anomalies and The Need for Perpective Investment Analyst, 91: 25-30. KEANE, S. M. 1989. Seasonal Market Anomalies and the Need for Perspective. Investment Analyst 91: 25-30. KEIM, D. B. 1983. Size Related Anomalies and Stock Return Seasonality: Further Evidence. /. Finan, Earns. 12: 13-32. KEIM, D. B. 1985. Dividend Yields and Stock Returns: Implications of Abnormal January Returns. / Finan. Earns. 14: 473-489. KEIM, D. B. and R. STAMHAUGH. 1984. A Further Investigation of the Weekend Effect in Stock Returns. /. Finan. : 819-837. REINGANUM, M. R. 1983. The Anomalies in Stock Behavior of Small Firms in January: Empirical Tests for Tax Loss Selling Effects./ Finan. Earns. 12: 89-104, ROGALSKI, R. and S.M. TlNlC. 1986. The January Size Effect: Anomaly or Risk Measurement. Finan. Analy. J. 42: 63-70. Roi i, R. 1982. A Possible Explanation of the Small Firm Effect. /. Finan. 36: 879-888. WONG, P.L., S. K. NEOH, K.H. LEE and T. S. THONG.

1989. Seasonality in The Malaysian Stock Market. Proceedings of the Inaugural International Conference on Asian-Pacific Financial Markets, National University of Singapore. YON(., O. 1989. Seasonality Effect in the Kuala Lumpur Stock Exchange. Malaysia Management Review 24: 37-43.

PERTANIKAVOL. 15 NO. 1, 1992

(Received 14 February 1991)

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