TOC Alerts Journal of Research in Biology Volume 4 Issue 4 PDF

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Abiya Chelliah [Molecular Biology] Dr. Afreenish Hassan [Microbiology]
Publisher, Journal of Research in Biology. Department of Pathology, Army Medical College, Rawalpindi, Pakistan.
Editorial Board Members: Gurjit Singh [Soil Science]

Ciccarese [Molecular Biology] Krishi Vigyan Kendra, Amritsar, Punjab, India.
Universita di Bari, Italy.
Dr. Marcela Pagano [Mycology]
Sathishkumar [Plant Biotechnologist] Universidade Federal de São João del-Rei, Brazil.
Bharathiar University.
Dr.Amit Baran Sharangi [Horticulture]
SUGANTHY [Entomologist] BCKV (Agri University), West Bengal, INDIA.
TNAU, Coimbatore.
Dr. Bhargava [Melittopalynology]
Elanchezhyan [Agriculture, Entomology] School of Chemical & Biotechnology, Sastra University, Tamilnadu, INDIA.
TNAU, Tirunelveli.
Dr. Sri Lakshmi Sunitha Merla [Plant Biotechnology]
Syed Mohsen Hosseini [Forestry & Ecology] Jawaharlal Technological University, Hyderabad.
Tarbiat Modares University (TMU), Iran.
Dr. Mrs. Kaiser Jamil [Biotechnology]
Dr. Ramesh. C. K [Plant Tissue Culture] Bhagwan Mahavir Medical Research Centre, Hyderabad, India.
Sahyadri Science College, Karnataka.
Ahmed Mohammed El Naim [Agronomy]
Kamal Prasad Acharya [Conservation Biology] University of Kordofan, Elobeid-SUDAN.
Norwegian University of Science and Technology (NTNU), Norway.
Dr. Zohair Rahemo [Parasitology]
Dr. Ajay Singh [Zoology] University of Mosul, Mosul,Iraq.
Gorakhpur University, Gorakhpur
Dr. Birendra Kumar [Breeding and Genetic improvement]
Dr. T. P. Mall [Ethnobotany and Plant pathoilogy] Central Institute of Medicinal and Aromatic Plants, Lucknow, India.
Kisan PG College, BAHRAICH
Dr. Sanjay M. Dave [Ornithology and Ecology]
Ramesh Chandra [Hydrobiology, Zoology] Hem. North Gujarat University, Patan.
S.S.(P.G.)College, Shahjahanpur, India.
Dr. Nand Lal [Micropropagation Technology Development]
Adarsh Pandey [Mycology and Plant Pathology] C.S.J.M. University, India.
SS P.G.College, Shahjahanpur, India
Fábio M. da Costa [Biotechnology: Integrated pest control, genetics]
Hanan El-Sayed Mohamed Abd El-All Osman [Plant Ecology] Federal University of Rondônia, Brazil.
Al-Azhar university, Egypt
Marcel Avramiuc [Biologist]
Ganga suresh [Microbiology] Stefan cel Mare University of Suceava, Romania.
Sri Ram Nallamani Yadava College of Arts & Sciences, Tenkasi, India.
Dr. Meera Srivastava [Hematology , Entomology]
T.P. Mall [Ethnobotany, Plant pathology] Govt. Dungar College, Bikaner.
Kisan PG College,BAHRAICH, India.
P. Gurusaravanan [Plant Biology ,Plant Biotechnology and Plant Science]
Mirza Hasanuzzaman [Agronomy, Weeds, Plant] School of Life Sciences, Bharathidasan University, India.
Sher-e-Bangla Agricultural University, Bangladesh
Dr. Mrs Kavita Sharma [Botany]
Mukesh Kumar Chaubey [Immunology, Zoology] Arts and commerce girl’s college Raipur (C.G.), India.
Mahatma Gandhi Post Graduate College, Gorakhpur, India.
Suwattana Pruksasri [Enzyme technology, Biochemical Engineering]
N.K. Patel [Plant physiology & Ethno Botany] Silpakorn University, Thailand.
Sheth M.N.Science College, Patan, India.
Dr.Vishwas Balasaheb Sakhare [Reservoir Fisheries]
Kumudben Babulal Patel [Bird, Ecology] Yogeshwari Mahavidyalaya, Ambajogai, India.
Gujarat, India.
CHANDRAMOHAN [Biochemist] Dr. Pankaj Sah [Environmental Science, Plant Ecology]

College of Applied Medical Sciences, King Saud University. Higher College of Technology (HCT), Al-Khuwair.
B.C. Behera [Natural product and their Bioprospecting] Dr. Erkan Kalipci [Environmental Engineering]
Agharkar Research Institute, Pune, INDIA. Selcuk University, Turkey.
Kuvalekar Aniket Arun [Biotechnology] Dr Gajendra Pandurang Jagtap [Plant Pathology]

Lecturer, Pune. College of Agriculture, India.
Mohd. Kamil Usmani [Entomology, Insect taxonomy] Dr. Arun M. Chilke [Biochemistry, Enzymology, Histochemistry]
Aligarh Muslim university, Aligarh, india. Shree Shivaji Arts, Commerce & Science College, India.
Dr. Lachhman Das Singla [Veterinary Parasitology] Dr. AC. Tangavelou [Biodiversity, Plant Taxonomy]
Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India. Bio-Science Research Foundation, India.
Vaclav Vetvicka [Immunomodulators and Breast Cancer] Nasroallah Moradi Kor [Animal Science]
University of Louisville, Kentucky. Razi University of Agricultural Sciences and Natural Resources, Iran
José F. González-Maya [Conservation Biology] T. Badal Singh [plant tissue culture]

Laboratorio de ecología y conservación de fauna Silvestre, Panjab University, India
Instituto de Ecología, UNAM, México.
Dr. Kalyan Chakraborti [Agriculture, Pomology, horticulture] Dr. Satish Ambadas Bhalerao [Environmental Botany]
AICRP on Sub-Tropical Fruits, Bidhan Chandra Krishi Viswavidyalaya, Wilson College, Mumbai
Kalyani, Nadia, West Bengal, India.
Rafael Gomez Kosky [Plant Biotechnology]
Dr. Monanjali Bandyopadhyay [Farmlore, Traditional and indigenous Instituto de Biotecnología de las Plantas, Universidad Central de Las Villas
practices, Ethno botany]
V. C., Vidyasagar University, Midnapore. Eudriano Costa [Aquatic Bioecology]
IOUSP - Instituto Oceanográfico da Universidade de São Paulo, Brasil
M.Sugumaran [Phytochemistry]
Adhiparasakthi College of Pharmacy, Melmaruvathur, Kancheepuram District. M. Bubesh Guptha [Wildlife Biologist]
Wildlife Management Circle (WLMC), India
Prashanth N S [Public health, Medicine]
Institute of Public Health, Bangalore. Rajib Roychowdhury [Plant science]
Centre for biotechnology visva-bharati, India.
Tariq Aftab
Department of Botany, Aligarh Muslim University, Aligarh, India. Dr. S.M.Gopinath [Environmental Biotechnology]
Acharya Institute of Technology, Bangalore.
Manzoor Ahmad Shah
Department of Botany, University of Kashmir, Srinagar, India. Dr. U.S. Mahadeva Rao [Bio Chemistry]
Universiti Sultan Zainal Abidin, Malaysia.
Syampungani Stephen
School of Natural Resources, Copperbelt University, Kitwe, Zambia. Hérida Regina Nunes Salgado [Pharmacist]
Unesp - Universidade Estadual Paulista, Brazil
Iheanyi Omezuruike OKONKO
Department of Biochemistry & Microbiology, Lead City University, Mandava Venkata Basaveswara Rao [Chemistry]
Ibadan, Nigeria. Krishna University, India.
Sharangouda Patil Dr. Mostafa Mohamed Rady [Agricultural Sciences]

Toxicology Laboratory, Bioenergetics & Environmental Sciences Division, Fayoum University, Egypt.
National Institue of Animal Nutrition
and Physiology (NIANP, ICAR), Adugodi, Bangalore. Dr. Hazim Jabbar Shah Ali [Poultry Science]
College of Agriculture, University of Baghdad , Iraq.
Jayapal
Nandyal, Kurnool, Andrapradesh, India. Danial Kahrizi [Plant Biotechnology, Plant Breeding,Genetics]
Agronomy and Plant Breeding Dept., Razi University, Iran
T.S. Pathan [Aquatic toxicology and Fish biology]
Department of Zoology, Kalikadevi Senior College, Shirur, India. Dr. Houhun LI [Systematics of Microlepidoptera, Zoogeography, Coevolution,
Forest protection]
Aparna Sarkar [Physiology and biochemistry] College of Life Sciences, Nankai University, China.
Amity Institute of Physiotherapy, Amity campus, Noida, INDIA.
María de la Concepción García Aguilar [Biology]
Dr. Amit Bandyopadhyay [Sports & Exercise Physiology] Center for Scientific Research and Higher Education of Ensenada, B. C., Mexico
Department of Physiology, University of Calcutta, Kolkata, INDIA .
Fernando Reboredo [Archaeobotany, Forestry, Ecophysiology]
Maruthi [Plant Biotechnology] New University of Lisbon, Caparica, Portugal
Dept of Biotechnology, SDM College (Autonomous),
Ujire Dakshina Kannada, India. Dr. Pritam Chattopadhyay [Agricultural Biotech, Food Biotech, Plant Biotech]
Visva-Bharati (a Central University), India
Veeranna [Biotechnology]
Dept of Biotechnology, SDM College (Autonomous),
Ujire Dakshina Kannada, India.
RAVI [Biotechnology & Bioinformatics]

Department of Botany, Government Arts College, Coimbatore, India.
Sadanand Mallappa Yamakanamardi [Zoology]

Department of Zoology, University of Mysore, Mysore, India.
Anoop Das [Ornithologist]

Research Department of Zoology, MES Mampad College, Kerala, India.
Table of Contents (Volume 4 - Issue 4)
Serial No Accession No Title of the article Page No
1 RA0446 Laboratory evaluation and comparative study of herbal mosquito coils 1332-1337
against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae).
Susheela P and Radha R.
2 RA0447 Daily Activity Budget of Nicobar Long-tailed Macaque (Macaca 1338-1347

fascicularis umbrosa) in Great Nicobar Island, India..
Rajeshkumar S, Raghunathan C, Kailash Chandra and Venkataraman K.
3 RA0454 Analysis on protein fingerprint, RAPD and fruit quality of tomato 1348-1356
mutants by ion beam implantation.
Duan HY, Wang CF, Yu YA, Li XW and Zhou YQ.
4 RA0452 The leaping behavior of the sally lightfoot crab Grapsus grapsus 1357-1364
(Crustacea: Decapoda: Brachyura) at an oceanic archipelago.
Marina de Sá Leitão Câmara de Araújo.
Journal of Research in Biology ISSN No: Print: 2231 –6280; Online: 2231- 6299
An International Scientific Research Journal
Original Research
Laboratory evaluation and comparative study of herbal mosquito coils

against the filarial vector, Culex quinquefasciatus (Diptera: Culicidae)
Authors: ABSTRACT:
Journal of Research in Biology
Susheela P* and Radha R.

Synthetic insecticides employed for the control of insect pests are toxic to
man and livestock acting as pollutants to the environment, killing all beneficial insects
thereby causing a disturbance to the ecosystem. The use of natural products such as
plant essential oils has assumed significance as an important component of insect pest
management because of their financial viability and eco-friendly nature. They hold
Institution:
promise as alternatives to chemical insecticides to reduce pesticide load in the
Department of Zoology,
PSGR Krishnammal College environment. A laboratory experiment was conducted to investigate the efficacy of
for Women Coimbatore, three essential oils -eucalyptus oil, lemon grass oil and thyme oil for the repellent
Tamilnadu, India. activity against the filarial vector, Culex quinquefasciatus. Among the essential oils,
Lemon grass oil showed good repellency property when compared to the other two
plant oils. Hence, the results of the investigation would indicate a significant potential
for lemon grass oil as a possible source of natural products that could be used as an
alternative to synthetic insecticides.
Corresponding author: Keywords:

Susheela P. Mosquito, Culex quinquefasciatus, repellency, Plant essential oil.
Article Citation:
Web Address:
http://jresearchbiology.com/ Susheela P and Radha R.
documents/RA0446.pdf. Laboratory evaluation and comparative study of herbal mosquito coils against the
filarial vector, Culex quinquefasciatus (Diptera: Culicidae)
Journal of Research in Biology (2014) 4(4): 1332-1337
Dates:
Received: 01 April 2014 Accepted: 31 May 2014 Published: 20 Jun 2014
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/

licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and
reproduction in all medium, provided the original work is properly cited.
1332-1337 | JRB | 2014 | Vol 4 | No 4

An International
Scientific Research Journal www.jresearchbiology.com
Susheela and Radha, 2014
INTRODUCTION contain active materials that are potential enough to

Mosquitoes are considered as an important insect control the mosquito population. (Sutthanont et al.,
pests that affect the health and well being of human 2010). In recent times, plant products are used as novel
beings and other animals worldwide. Mosquitoes are chemo therapeutants in pest management in different
cosmopolitan in distribution and have occupied many parts of the world, because of their biodegradable nature.
niches including higher altitudes. Mosquitoes are always (Hardin and Jackson, 2009).Therefore, the present study
considered as a nuisance because they consume blood was aimed to investigate the mosquito repellent nature of
from living vertebrates, including human beings three essential oils: Eucalyptus tereticornis (Eucalyptus),
(Bernhard et al., 2003). In India, annually around 40 Cymbopogon citratus (lemon grass) and Thymus vulgaris
million people suffer from mosquito borne diseases. The (thyme) against C. quinquefasciatus.
extensive use of mosquito repellents and insecticides in
public health programmes has caused extensive level of MATERIAL AND METHODS
environmental pollution and serious health hazards. Plant Oils:
Many of them are alarmingly toxic to human beings and The plant oils were purchased from the Aromatic
also other non-target organisms. Oil Stores, Coimbatore, Tamil Nadu and formulated for
Controlling the mosquitoes in an effective manner is the experiment. A stock solution at 1000 ppm is prepared
often complex and expensive task which requires support by dissolving the essential oils in distilled water using
from communities and also from different groups such as 2 ml of 100% acetone respectively. The serial dilutions
industry, agriculture, state and local governments of essential oils at the concentration of 5%, 15% and
(Joseph et al., 2004). The harmful effect of the pesticides 25% and three replicate of each concentration were
on the environment, animals, plants and human beings is made.
an issue of great concern. As far as India is concerned, Preparation of herbal mosquito coils:
many of the insecticides and larvicides are Mosquito coils were prepared using cow dung,
commercialized in the form of dust, powder or sprays sawdust, neem leaves, flower waste and tulsi leaves.
that contain chemicals such as organochlorine, Then the essential oils, Thymus vulgaris, Lemon grass,
organophosphates and synthetic pyrethroid. Yet and Eucalyptus oils were sprayed (w/w) on top of the
mosquitoes, due to a prolonged use of these insecticides coil by using a hand spray pump in different
become resistant and thus it becomes a difficult task to concentration of 5%, 15% and 25 % separately and they
eradicate them totally (Prajapati et al., 2005). They also were used for its efficacy against C. quinquefasciatus
pose a threat to the human population by carrying vector mosquito. The coil was dried in the oven at 70°C for
borne diseases and sometimes out break as epidemics. 6 hours was dried for half an hour at room temperature.
Hence to control the vector mosquitoes, efforts are These coils were then packed in suitable air tight plastic
being taken to look for an alternate solution which folders and kept for 2 – 3 days for even spread of the
will ultimately minimize the use of synthetic essential herbals on the coil.
insecticides. Test Organisms
The development of eco-friendly insecticides will The test organism, C. quinquefasciatus, was reared in
serve its purpose as a new alternate to substitute the the laboratory in the Department of Zoology, PSGR
synthetic insecticides essentially cutting down the Krishnammal College for Women, Coimbatore, Tamil
chemical pollution. The pyrethrum flower extracts Nadu. Dog biscuits and yeast powder in a ratio of 3:1
1333 Journal of Research in Biology (2014) 4(4): 1332-1337
Concentration of lemon grass oil

Figure-1 Repellency of lemon grass oil against C. quinquefasciatus
were given as feed for the mosquito larvae. On the other coil was put in the middle of one side of the room. For
hand, adult mosquitoes were fed with a 10% sucrose control, 50 female unfed, 5 days old mosquitoes were
solution and a 1 week-old chick. Mosquitoes were kept released in the centre of the room. Then the number of
at relative humidity of 28-30°C, 75 ± 5%, with 14-h light landing mosquitoes on the bare legs of the human
and 10-h dark, photo period ( Kitzmiller et al., 1954). volunteers was counted for a period of 2 min. For testing,
Bioassays the mosquito coil was ignited, then counting of the
Repellency Test number of landing mosquitoes on the bare legs of the
The experiment was conducted in a closed room, human volunteers began and was recorded at periodic
with a volume of 92.8 m3 in the Department of Zoology, intervals. Three replications were done by changing the
PSGR Krishnammal College for Women, Coimbatore, positions of the human volunteers, and then repeating the
Tamil Nadu. The human volunteers sat at 1 m, 2 m, 4 m, procedure the next day.
and 8 m from the herbal mosquito coil. The mosquito
Concentration of eucalyptus oil

Figure-2 Repellency of eucalypus oil against C. quinquefasciatus
Journal of Research in Biology (2014) 4(4): 1332-1337 1334

Concentration of thyme oil
Figure-3 Repellency of thyme oil against C. quinquefasciatus
RESULTS AND DISCUSSION A number of studies have been focused on lemon

The results of repellency test of thyme oil against grass oil for controlling mosquitoes as a larvicide and a
C. quinquefasciatus (Say) after one hour of treatment are repellent with varied results. Hanifah et al., (2011)
presented in Figure-3. The results clearly indicated that demonstrated C. citratus extract has more acaricidal
the highest repellency was reported at 25% concentration activity against Der-matophagoides farina and
of thyme oil when compared to 5% concentration and D. pteronyssinus than Azadirachta indica at 50%
10% concentration. As the concentration of the plant oil concentra-tion. This proves the efficiency of
formulation increases the total mortality of Cymbopogon citratus in controlling the insect pests.
C. quinquefasciatus also gets increased. Figure-2 Oyedele et al., (2002) evaluated the ointment and cream
revealed the efficacy of eucalyptus oil against formulations of lemon grass oil in different classes of
C. quinquefasciatus. The lowest repellency was observed base and the oil in liquid paraffin solution for mosquito
at 5% concentration of eucalypus oil and the highest repellency in a topical application. Cilek et al., (2011)
repellency was observed at 25% concentration. But the studied the efficacy of several commercially formulated
essential oil, eucalypus oil is more effective than thyme essential oils against caged female Aedes albopictus and
oil. Increase in the concentration of the plant oil Culex quinquefasciatus. Mgbemena (2010) found that
formulation was found to increase the total repellency of the essential oil O. gratissimium had a greater larvicidal
Culex quinquefasciatus. The different concentrations of activity than C. citratus. Purwal et al., (2010) tested the
the lemon grass oil was recorded against activity of C. citratus and Mentha piperita essential oils
Culex quinquefasciatus in Figure-1. The percentage of in a combination against Pe-diculus humanus and found
repellency was found to be high in 25 % concentration a mean time to death of 60 minutes. Therefore the
than 5 % concentration of the plant oil. The results of essential oils can be used as an alternative to synthetic
this study clearly indicated that lemon grass oil had high insecticides for vector control programmes.
repellency potential to control the mosquitoes than the The essential oils (EO) eucalyptus oil, lemon
other two essential oils. grass oil, thyme oil were evaluated for repellent activity
against the Culex quinquefasciatus. Essential oils of
many plants were observed to have mosquito larvicidal Hanifah AL, Awang SH, Ming HT, Abidin SZ and
property and have received attention as potentially Omar MH. 2011. Acaricidal activity of Cymbopogon
controlling vectors of mosquito borne disease (Zhu et al., citratus and Azadirachta indica against house dust mites.
2006). Therefore, the use of lemon grass oils in insect/ Asian Pac J Trop Biomed., 1(5):365-369.
mosquito control is an alternative pest control method for
Hardin JA and Jackson FLC. 2009. Applications of
minimizing the harmful effects of pesticidal compounds
natu-ral products in the control of mosquito-transmitted
on the environment. The present study has identified
diseases. Afr J Biotechnol., 8(25): 7373-8.
more plant oils showing larvicidal activity against
Culex mosquito. The results obtained suggest that the Joseph CC, Ndoile MM, Malima RC and Nkuniya
plant oils are promising as larvicides against MH. 2004. Larvicidal and mosquitocidal extracts, a
Culex mosquito. The present study also suggests the use coumrin, isoflavonoids and pterocarpans from
of Lemon grass oil as the most effective alternative in Neorautanenia mitis. T Roy Soc Trop Med H., 98(8):
controlling mosquitoes. 451-455.
Kitzmiller JB and Micks DW. 1954. Techniques for

CONCLUSION
rearing Culex mosquitoes. Am. Midland Nat., 52(1): 253
The results of the present investigation proved
-256.
that the all essential oils at higher concentration are
effective but lemongrass oil exhibit a significant knock Mgbemena IC. 2010. Comparative evaluation of
down activity at higher concentration when compared to larvicidal potentials of three plant extracts on Aedes
the other oils. For the commercialization of these herbal aegypti. J Am Sci., 6: 435-40.
mosquito coils, further simulated and actual field trials
Oyedele AO, Gbolade AA, Sosan MB, Adewoyin FB,
are required. Hence, Lemongrass essential oil, alone or in
Soyelu OL and Orafidiya O. 2002, Formulation of an
combinations with those obtained from other mosquito
Effective Mosquito-repellent Topical Product from
repellent plant species, could be potentially used for the
Lemongrass Oil, Phytomedicine. 9(3):259-262.
preparation of mosquito repellent products.
Prajapati V, Tripathi AK, Aggarwal KK and
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Original Research
Daily Activity Budget of Nicobar Long-tailed Macaque

(Macaca fascicularis umbrosa) in Great Nicobar Island, India
Authors: ABSTRACT:
Rajeshkumar S1*,
Raghunathan C1, Nicobar long-tailed macaques (Macaca fascicularis umbrosa Miller, 1902) are
Kailash Chandra2 and distributed in three Islands of Nicobar namely Great Nicobar, Little Nicobar and
Venkataraman K2. Katchal. Their insular population requires special attention from research and
management perspectives. Daily activity budget of M.f. umbrosa in the Great Nicobar
Institution: Island was studied from October 2011 to September 2013 by intensive direct
1. Zoological Survey of observation method. Study revealed that Nicobar long-tailed macaque, undergoes
India, Andaman and Nicobar most of the time for Locomotion (36.07%), followed by feeding (22.35%), resting or
Regional Centre, Port Blair- being inactive (15.74%), grooming (11.14%), vocalization (7.03%), playing (5.64%),
744 102, Andaman and sexual arousal (1.46%) and agonistic (0.56%). All daily activities have significant
Nicobar Islands, India. difference (χ2 = 1156.22; df = 7, P = 0.05). Chi-square test demonstrated that the daily
activity budget differed significantly among the behaviours. Qualitative results found
2. Zoological Survey of that the interaction within the group was fighting and grabbing food. The significant
India, M-Block, New
observation of disability in their legs was noticed in Nicobar Long-tailed Macaque. The
Alipore, Kolkatta-700 053,
relation between their behaviour and disability is also discussed.
India.

Rajeshkumar S. Macaca fascicularis umbrosa, Daily activity budget, Great Nicobar Island
Article Citation:
Email Id:
Rajeshkumar S, Raghunathan C, Kailash Chandra and Venkataraman K.
Daily Activity Budget of Nicobar Long-tailed Macaque (Macaca fascicularis umbrosa) in
Great Nicobar Island, India.
Web Address: Dates:

http://jresearchbiology.com/ Received: 01 Apr 2014 Accepted: 30 May 2014 Published: 24 Jun 2014
documents/RA0447.pdf.
1338-1347 | JRB | 2014 | Vol 4 | No 4

An International
Rajeshkumar et al., 2014
INTRODUCTION Veira, 2002; Hamada et al., 2008). Previous researches

Primates are maintaining the sustainable in Nicobar subspecies are available for population status
ecosystem and play as indicator for ecosystem health; and distribution profiling (Umapathy et al., 2003;
hence, they help in making of conservation and Sivakumar, 2010; Narasimmarajan and Raghunathan,
management plans. Non-human primates of undisturbed 2012) Study on ecology and behaviour are also focused
areas are having great behavioural variation (Thomas, in the other subspecies of Long-tailed Macaque in South
1991) which are closely related to human beings such as East Asian countries. Reports are available on the
eating, playing, fighting, keeping young ones etc. (Rod aggressive and social behaviour of M. fascicularis
and Preston-Mafham, 1992). The daily activities and (Nordin and Jasmi, 1981; Zamzarina, 2003; Brent and
behaviour of primates differ between residential, non- Veira, 2002; Khor, 2003; Md-Zain et al., 2003; Siti,
residential and undisturbed areas (Krebs and Davies, 2003). The present study is focused on the daily activity
1993). Large group size, poor habitat quality, seasonal budgets of M f. umbrosa in Great Nicobar Island.
variation in food availability may affect their daily
activity budget (Peres, 1993; Passamani, 1998). The MATERIALS AND METHODS
Long-tailed macaques (Macaca fascicularis umbrosa Study Area
Miller, 1902) are the only non-human primates found on The Great Nicobar Island is about 1045.1 sq km
Nicobar Islands (Umapathy et al., 2003). Other comprises of Campbell bay National Park and Galathea
subspecies occur in Myanmar, Cambodia, Laos, National Park (Fig. 1). These two National Parks
Vietnam, Thailand, Malaysia, Indonesia and the embrace Great Nicobar Biosphere Reserve (GNBR). The
Philippines (Rodman, 1991; Tikader and Das, 1985). study site covers about 3 km2 and is composed of low
This species varies in their behaviour, social hills near dense semi evergreen forest, Maggar Nallah
organisations, habitat consumption, morphology and river and human Settlements at Govind Nagar (06°
genetic variation due to wide distribution (Brent and 59.985' N 093° 54.459' E) and it is 6 km away from
Campbell Bay (Fig. 1). GNBR has richest faunal and
Fig 1. Study area and Study site.

floral communities. Great Nicobar is the home for plants grooming, vocalization, playing, sexual arousal and
like Albizia chinensis, Albizia lebbeck, Artocarpus agonistic were collected during the study. Chi-square test
chaplasha, Calophyllum soulattri, Dipterocarpus sp., was applied to analyse the behaviour data set obtained.
Pterocarpus sp., and Sterculia campanulatum. In fauna, The nonparametric χ2 test was used to analyze the
other than the long-tailed macaques, the endemic significance of activity budgets.
mammals recorded are Nicobar wild boar (Sus scrofa
nicobaricus), Nicobar Tree shrew (Tupaia nicobarica), RESULTS AND DISCUSSION
Nicobar shrew (Crocidura nicobarica) and Nicobar Result on the percentage of eight daily activities
Flying fox (Pteropus faunulus). of Nicobar long-tailed macaques monitored is given in
Behaviour Sampling Method Table 2. Chi-square analysis upon the present study
Following the methods of Hambali et al., (2012); indicated that all the eight behavioural observation shows
Md-Zain et al., (2008b) and Brent and Veira (2002) daily significant differences (Table 2). Jaman and Huffman
activity observations of macaque were made during 2 to (2008) observed that, activities of Japanese macaque
3 days in a week at 0500 hours until 1630 hours for 78 (M. fuscata) in captivity varied between age-sex classes.
days from October 2011 to September 2013 to determine Similarly the behavioural variation occurred in
the behaviour categories. A study group categories and individual with different age-sex observed in the present
its composition of the three consecutive years are given study. The most observed daily activity for all the age
in Table. 1. The total number of individuals in study group was locomotion. The locomotion is the highest
group increased year by year i:e from 37 to 51 portion of daily activity in long-tailed macaques
individuals. Every year the numbers of females were compared to other activities (Hambali et al., 2012; Md-
more than that of males. This group was marked by their Zain et al., 2010; Sia, 2004; Suhailan, 2004). This is
dominant male who had a distinctive large and elongated because of diurnal in nature as they are very active
white area between the eyes and white eyelids compared during the day as they use their maximum time in
to the other groups. Focal animal sampling method was searching for food.
adopted to collect the quantitative data at ten minutes Locomotion
interval (Altmann, 1974; Lehner, 1979). During According to Menard (2004) and Wheatley
torrential rain and adverse weather condition, the (1980) Long-tailed Macaques are the primates spending
observation was discontinued until the weather resumes most of their time for moving as they are mainly
normally, because the animals were partially obscured or frugivorous and occupy more space. It was also observed
moved completely from the observation sites. The data that the study group’s moving choice is varied day by
on the observations of locomotion, feeding, resting, day to different location and range. When they move out
Table 1. Year wise group composition and total number of Individuals in the study group.
Group categories Adult (Mature) Immature
Total No. of Individual
Year Male Female Total Sub Adult Juvenile Infant
2011 (October) 10 13 23 10 3 1 37
2012 (March) 12 15 27 12 4 2 45
2013 (August) 13 16 29 12 6 4 51

Table 2. Percentage and Chi-square value of Nicobar long-tailed macaque’s daily activity.
Activity Observation Percentage (%) Expected frequency χ2 = (O-E)2/E

Locomotion 518 36.07 179.5 638.34*
Feeding 321 22.35 179.5 111.54*
Resting 226 15.74 179.5 12.04*
Grooming 160 11.14 179.5 2.12*

Vocalization 101 7.03 179.5 34.33*
Playing 81 5.64 179.5 54.05*
Sexual 21 1.46 179.5 139.95*
Agonistic 08 0.56 179.5 163.85*
Total 1436 100 1436 1156.25
* Showing significant differences (p<0.05), by using the chi-square test (χ2).

Degrees of freedom (df) = 7, O-Observation, E-Expected frequency.
of their home range, there was a shortage of food sources highest proportion of time in resting rather than feeding
and availability of fruits. According to O’Brien and depending on the food and weather factor. An increase in
Kinnaird (1997), availability of food source significantly one activity may pose some influence on other activities
affects their locomotion in daily activity pattern. (Jaman and Huffman, 2008). The main food sources are
Sometimes these animals visit human settlement areas fruits, flowers, tender leaves, insects, crabs, beetles,
and raids crop land, coconuts farms and banana farms butterflies, some spiders, grasshopper etc. Usually
which lead to their destruction. The result indicates that macaque feed insects in afternoon period between resting
the macaque spent most of the time in moving due to the and grooming. When the food sources are less long-
insufficient food sources in their habitat. Likewise this tailed macaque usually rest.
study group also spend most of their time to visit Resting
different localities because of their diminishing natural Resting is the third most activity observed in our
food sources. study (Fig. 2 B). The result of the study revealed that
Feeding prolonged feeding activity considerably reduced the
Besides locomotion, feeding was observed as one resting behaviour during the observation from macaque
of the major activities of macaque during the study (Fig. in Great Nicobar as noticed by Hambali et al., (2012) in
2 A). It resembles with the other subspecies studied by Malayan long-tailed macaque and Kurup and Kumar
Hambali et al., (2012), Md-Zain et al., (2010), Suhailan (1993) in lion-tailed macaque. Resting includes activities
(2004) and Tuan-Zaubidah (2003) who all found that like sleeping, lying down and to sit idle. Macaques were
feeding is the second most occurrence activity compared observed resting on tree branches, dead woods, bushes,
to other. However this finding was contradict with other rocks and sometimes resting on the roads. Also they use
macaque species. For example Southern India wild lion- to take a few minutes rest after walking continuously.
tailed macaque (Kurup and Kumar, 1993) and captive Rainy season and unusual climate directly affect their
Japanese macaque (Jaman and Huffman, 2008) spend the feeding and moving activities and increase their resting

activity. During night time, macaques sleep on the top of than social grooming. Social grooming highly noticed
tree branches. This behaviour indicates that the macaque between the adult female and adult male. Observations
protect themselves from the predators. The only known on grooming between the adult female with infants were
predator is reticulated python (Broghammerus least due to the presence of only few infant in the group.
reticulatus) as no other higher predators are found in There was a least observation on grooming between
Great Nicobar Island, but the anthropogenic activity and adult female and juveniles as well as sub adults. Self-
domestic predators like dogs also affects their normal grooming was also often observed in sub adults and
activity. secluded male at the time of resting. In addition, after
Grooming mating, the dominant male is groomed by female.
Grooming is the fourth highest activity observed According to Lazaro-perea et al., (2004) this behaviour
after resting (Fig. 2 C). This result is similar with M. can be a way to get protection from others while fighting
fascicularis found in Kuala Selangor Nature Park, and also for sharing of food.
Malaysia (Hambali et al., 2012). Most of their grooming Vocalization
activity occurs at the time of resting period. It was Vocalization is the fifth behaviour that has been
predominantly observed at late afternoon when the observed. When the agonistic interaction occurs between
macaques return to the home range. At the time of the group individuals, dominant adult male produce loud
grooming one monkey picks up lice from other’s body. calls and all the other individuals sound continuously. In
Most of the individuals often prefer to self-groom rather general, macaque produces loud calls especially for
Fig 2. Various activities of Long-tailed Macaque in Great Nicobar Island

A. Feeding, B. Resting, C. Grooming, D. Playing, E. Mating, F. Agonistic.

grabbing and snatching food item and fighting with their categories observed during the study. It was also
group member. In addition during agonistic interaction observed that these animals prefer playing on the
within the group or entrance of predatory animals such as selected trees like Casuarinas, Pandanus, Guava and
dogs in their territory, macaque used to make Coconut. In the evening, all the group member moves
vocalization. Normally vocalization can be treated as a near sleeping site and while moving many were found
warning signal to protect themselves from predators as collecting and eating some insects in the bushy area.
observed by Md-Zain et al., (2010). Due to the Sexual Arousal
observer’s or the human’s activity in their range, Sexual behaviour like mating, mount, inspect
macaque produce different sounds and mainly the sub copulation are the categories were observed as the
adults seem to be most active as they used to climb very seventh activity (Fig. 2 E). In our study period dominant
quickly and keep other individuals alert. Members of the males were actively involved in mating with adult
group after hearing the vocal call warning used to climb females as this may help females in giving birth to
to higher ground to escape or hide in bushes. We healthy generation. Females use to live with multimale
observed a least number of calls produced by macaques group, focused in copulating with dominant males as
while playing activity. Kipper and Todt (2002) and Md- observed by Hambali et al., (2012), Lawler et al., (1995),
Zain et al., (2010) also found that the vocal call was Md-zain et al., (2010) and Van Noordwijk and Van
produced by macaques while playing. In the present Schaik (1999). Sexual behaviour observed is only a small
study the male long-tailed macaques were found to portion of daily activity in long-tailed macaque.
produce vocal calls while grooming after mating. No Normally the adult male was found to smell or observe
females were observed producing vocals during mating. the adult female genitalia first to make sure that the
On the other hand observation made by Md-Zain et al., females are ready to mate or not which is in corroborated
(2010) showed that females were found to produce vocal with the report of Brent and Veira (2002), Md Zain et al.,
during and after mating. The possible reason for this (2010) and Hambali et al., (2012). The long-tailed
behaviour can be a hormonal effect (Engelhardt et al., macaque takes a few seconds for mating activity.
2005). Agonistic Activity
Playing The least observed activity is the agonistic
Playing activity is the sixth behaviour that has behaviour (Fig. 2 F). During our study chase, grab, hit,
been observed during the study period (Fig. 2 D). We bite and fight are the categories of agonistic behaviour
found predictable differences in playing activity in the observed as the eighth activity. Though these behaviours
juveniles and sub adults. Juveniles were found to play are supported by Hambali et al., (2012), Md-Zain et al.,
more than sub adults. Adult macaques were not involved (2010), Suhailan (2004) and Tuan-Zaubidah (2003) they
in playing activity. The playing behaviour may form a found that mating is the least observed activity. Fighting
social competition and juveniles in their active age behaviour occurred while gaining foods and mates.
period will learn on social relations (Kipper and Todt, Hambali et al., (2012) found that Malay wild long-tailed
2002). Usually, playing behaviour was observed in the macaque has a hierarchy in the group, so that they have
late afternoon, when adult long-tailed macaques are their own way to avoid fight when looking for food
inactive. Wrestling, chasing, tickling, swinging on the together. Chasing and biting occur sometime between the
tree branches, pulling their tails to play with one another males and sub adults. Adult male were more aggressive
and invert hanging and jumping were the playing when their food was grabbed by other males, this shows
that the aggression appeared in males higher than activities, etc. The relation between disability and
females which is agreed with the Brent and Veira (2002) behaviour is also reported in Japanese macaques
from macaque observed at Indo-China population. (Macaca fuscata) by Turner et al., (2012). The possible
Significantly we observed few aggressive activities in the causes of disabilities are congenital defects, dog chasing
Nicobar long-tailed macaque against human beings and anthropogenic activities. However, exact cause of
especially women and children during the study period. disability was not known. But this significant
Disability and Behaviour observation may throw some light on the threats and
During our study period several disabled their status of these monkeys.
macaques were spotted (Fig 3). They were not able to
move properly due to their disability. These disabilities CONCLUSIONS
may cause some changes in their daily activities which in The present study enlightened behavioural and
turn will cause changes in their behaviour like activity patterns of the long-tailed macaque population
locomotion, disability in finding mates, foraging living in the Great Nicobar Island. It is revealed that
Fig 3. Disability in Nicobar Long-tailed macaque

A. Forearm partially disabled, B. Foreleg disabled, C. Hindleg partially disabled.

locomotion, feeding and resting were the most common 70(12): 1133-1144.
daily activities of these monkeys. Disabled macaques
Kamarul Hambali, Ahmad Ismail and Badrul Munir
spotted during our study period may give some
Md-Zain. 2012. Daily Activity Budget of Long-tailed
information on the changes in their behaviour that occur
Macaques (Macaca fascicularis) in Kuala Selangor
due to disability as well as on the threats they use to
Nature Park. Int. J. Basic and Applied Sciences. 12(4):
encounter. This study also found that the aggressive
47-52.
behaviour against humans may raise the issue of human-
macaque conflict. Further studies on the specific impact Khor OP. 2003. Kajian kelakuan Macaca fascicularis
of crop raiding and feeding behaviour will derive the dan interaksi dengan manusia di Taman Belia, Pulau
implication of its conservation and management Pinang. Tesis sarjana muda, Universiti Kebangsaan,
strategies. Malaysia.
Kipper S and Todt D. 2002. The use of vocal signals in

ACKNOWLEDGMENTS
the social play of Barbary Macaques. Primates. 43(1): 3-
The authors are grateful to the Ministry of
17.
Environment and Forests, Government of India. The
logistic support provided by Divisional Forest officer, Krebs JR and Davies NB.1993. An introduction to
Nicobar Division, Campbell Bay is duly acknowledged. behavioural ecology. Wiley-Blackwell Scientific
Publications, London.
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Original Research
Analysis on protein fingerprint, RAPD and fruit quality of

tomato mutants by ion beam implantation
Authors: ABSTRACT:
Duan HY*, Wang CF, In this research, seeds of tomato were irradiated by ion beam or treated with
Yu YA, Li XW and ion beam and soybean DNA, and some tomato mutants with morphological variations
Zhou YQ.
were analyzed. Protein analysis in the leaves of mutants showed, changes of protein
pattern in mutants were different as compared with the control, the main variation of
protein pattern were darkening of bands, increase of protein bands were detected in
mutant 12, mutant 14 and mutant 15 and lose of a band in mutant 15. Genomic DNA
of mutants were analyzed by RAPD, and total number of amplification bands, number
of differential bands and rate of differential bands were studied among mutants.
Compared with the control, rate of differential bands was 100.0 % in mutant 9 and 15,
Institution: also high in mutant 14 and 12, but was 20.0-50.0 % in other mutants except for
College of Life Science, mutant 3 and 11 without differential bands. In addition, content of vitamin C, soluble
Henan Normal University, saccharide and protein were different, and fruit quality was multifarious in the fruit of
Xinxiang 453007, China. mutants compared with the control; mutant 7 has better comprehensive nutritional
quality of fruit, whereas mutant 12 and 14 stand second. The above results showed
that effects of ion beam or soybean DNA on tomato genomic DNA would lead to the
changes in gene expression, protein synthesis and fruit quality, moreover some
tomato plants with better fruit quality or special characters were achieved, which
would provide basis for the application of ion beam technology in tomato breeding.

Duan HY. Ion beam, tomato, SDS-PAGE, RAPD, fruit quality.
Email Id: Article Citation:

Duan HY, Wang CF, Yu YA, Li XW and Zhou YQ.
Analysis on protein fingerprint, RAPD and fruit quality of tomato mutants by ion beam
implantation.
Web Address:
http://jresearchbiology.com/
documents/RA0454.pdf. Dates:
Received: 03 Jun 2014 Accepted: 06 Jun 2014 Published: 26 June 2014

1348-1356 | JRB | 2014 | Vol 4 | No 4

An International
Duan et al., 2014
INTRODUCTION MATERIALS AND METHODS

In recent years, mutation breeding has been a Plant materials
novel way in plant genetic improvement, especially low In this study, seeds of tomato (tomato Zhongza
energy ion beam implantation which exhibits many No. 9) were provided by Vegetable Flower Institute of
advantages, such as low damage, high mutation rate, Agricultural Sciences, Beijing, P. R. China, and were
wide mutational spectrum, and so on (Yu, 2000). At respectively irradiated by N+ or Ar+ ion beam in the 30
present, ion beam mutation breeding technology has kev energy conditions. Seeds of soybean (soybean
been successfully applied to a lot of crop breeding, such Zaoshu No. 2) were preserved in our laboratory, soybean
as rice, wheat, tobacco, cotton, soybean, rape and others seedlings with single-leaf were used to extract genomic
(Zhou, 2009). In addition, the etching and sputtering DNA with CTAB method, and DNA fragments of
effects of ion beam on cells would be very beneficial to soybean genomic DNA were obtained by ultrasonication.
foreign DNA entering into the cells (Wang et al., 2009, Culture of tomato plants
Li and Sun, 2011) and some transgenic plants have been Tomato seeds implanted with N+ or Ar+ ion beam
achieved by ion beam implantation (Duan et al., 2012), were treated as described in research (Ji et al., 2001), at
thus the transgenic technology mediated by ion beam is a first were respectively immersed into 0.1×SSC buffer
simple and feasible transgenic method. solution or 300 µg ml-1 DNA working solution which
Tomato is one of the most important vegetables was composed of soybean DNA and 0.1×SSC buffer
and fruits that contain abundant nutrients, such as solution, and then were separately washed several times
lycopene, vitamin C, trace elements and other nutrients with sterile water, but the control was only immersed
(Xue et al., 2004, Wang et al., 2010). In order to meet into sterile water. The above seeds were sowed in the test
the need of people, germplasm resources or genetic field and cultured under the greenhouse conditions with
improvement breeding of tomato is required to be 20°C light and 10°C dark temperature cycle. Seven days
studied and new cultivar of tomato should be cultivated. later, seeds germinated, seedlings with two leaves were
+
In our laboratory, it was found that nitrogen ion (N ) or transplanted in nutritive bowl and continued to be
+
argon ion (Ar ) had obvious influences on cell mitosis cultured. When cultured for two months, seedlings with
and chromosome structure, and lead to various types of five or six leaves were transplanted in the test field and
chromosome aberrations (Duan et al., 2013). Thus, dry cultured at the above culture condition.
seeds of tomato (tomato Zhongza No. 9) were irradiated In addition, the variations of morphologic
+ +
by N or Ar ion beam and soak into soybean DNA after characters in tomato plant were found, such as tall plant,
ion beam implantation to obtain a series of new fat leaves, thick stalk, and so on, moreover protein and
germplasm and cultivars with important application DNA fingerprint of some tomato mutants were
value, and some tomato mutants with the variations of respectively analyzed by SDS-PAGE or RAPD, and
morphologic characters were found in M1 present several indexes of fruit quality were also detected.
generation. In this research, tomato mutants with SDS-PAGE of protein in leaves
morphologic variations were analyzed by SDS-PAGE Proteins were extracted from the fresh leaves of
and RAPD, and several indexes of fruit quality were also tomato plants with morphologic variations as described
detected, which would provide foundation for new previously (Ji et al., 2001) with modifications. 1.0 g
cultivars of tomato and theoretical basis for ion beam leaves were mixed together with 1ml sterile water and
mutation breeding of tomato. grinded in the mortar on ice-bath, and then the
Duan et al., 2014
homogenate of leaves were centrifuged for 20 min by the test tube was sealed with plastic film and put in
12000 rpm at 4°C. The supernatant in the centrifuge tube boiling water for 30 min to extract soluble saccharide.
was transferred to 5 ml volumetric flask, furthermore, the The crude extract was filtered into 10 ml volumetric
precipitate in the centrifuge tube was extracted again flask, simultaneously the text tube and residues were
with sterile water and then the supernatant was also rinsed repeatedly with sterile water, and then the extract
transferred to the above 5 ml volumetric flask, in which was diluted with sterile water to constant volume. The
the supernatant was diluted with sterile water to a content of soluble saccharide was determined with
constant volume, then the solution was mixed and spectrophotometry at 485 nm, and the standard curve of
preserved at -20°C. The content of soluble protein in the soluble saccharide was drawn with sucrose. In addition,
above solution was determined by Bradford colorimetric determination of soluble saccharide content was repeated
method (Bradford, 1976) at 595 nm, and the standard three times.
curve of soluble protein was drawn with Bovine Serum Determination of vitamin C and protein in fruit
Albumin (BSA). In this research, SDS-PAGE of protein Mature fruits of tomato mutants were crushed
was performed under experiment conditions of 3 % with juicer, 0.5 g tomato juices were diluted with sterile
stacking gel (pH6.8), 12 % separating gel (pH8.8) and water to 100 ml volumetric flask, then extracted by
Tris-Glycine buffer solution (pH8.3), and Coomassie vacuum extrusion machine and preserved for the
Brilliant Blue method was used in this research. determination of fruit protein and vitamin C.
RAPD amplification Determination of fruit protein was performed as
In this study, leaves of tomato mutants were used determination of leaf protein in tomato, content of
to extract DNA by CTAB extraction procedure (Ausubel vitamin C was assayed by spectrophotometry (Chen
et al., 1987). RAPD amplification was performed as the et al., 2012) with modification and the standard curve of
method (Kangfu et al., 1994). Reaction system of RAPD vitamin C was drawn with standard vitamin C.
amplification was 25 μl and composed of 20 ng DNA, Moreover, determination of vitamin C and protein was
-1 -1
0.2 μmol L primer, 0.2 μmol/L dNTPs, 2.0 mmol L repeated three times.
Mg2+, 1U Taq DNA polymerase and double distilled
water. RAPD amplification was performed as follows: RESULTS AND DISCUSSION
initial denaturalization at 94°C for 5 min, followed by 35 Protein fingerprint in the leaves of tomato mutants
cycles of 94°C for 1 min, 36°C for 1 min and 72°C for It is well known that, effects of ion beam on
1.5 min, with a final extension cycle of 72°C for 8 min. plant are very obvious and could cause versatility, such
In addition, 100 primers were screened to obtain primers as stem diameter, flowering phase, plant height, quality
by which amplification bands are most distinctive, characteristic, and so on (Phanchaisri et al., 2012). In this
numbers of amplification bands are more and the research, protein in the leaves of tomato mutants were
repeatability is preferable. analyzed by SDS-PAGE (Figure 1), and the electro
Determination of soluble saccharide in fruit photograph was drawn in Figure 2 to more clearly
Assay of soluble saccharide was performed by observe changes of the protein pattern. As compared
enthrone colorimetric method (Liu et al., 2013) with with the control, the main variation of protein pattern in
improvement. Mature fruit of tomato mutants was the mutants were some bands darkening, especially the
crushed with juicer, 0.5 g tomato juices were mixed band with 0.350 Rf value obviously darkened, however
together with 5 ml sterile water in test tube, subsequently lose and increase of protein band was less found, only

Duan et al., 2014
Figure 1: Protein pattern in the leaves of tomato mutants by SDS-PAGE
M: marker, 1: the control, 2-11: tomato mutant induced by ion beam and soybean
DNA, 12: tomato mutant induced with 2×1017N+/cm2 ion beam, 13: tomato mutant
induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced with 2×1017Ar+/cm2
ion beam, 15: tomato mutant induced with 4×1017Ar+/cm2 ion beam.
two bands increased in mutant 12, mutant 14 and mutant curcas (Pamidimarri et al., 2010), Balsamine (Gao et al.,
15, and the Rf values were 0.05 and 0.083 respectively, 2012), and so on. In this research, genomic DNA of
furthermore mutant 15 lost one band (Rf=0.133) in tomato mutants was also analyzed with RAPD markers
comparison with the control and other mutants. The in order to explore changes in the genomic DNA.
above results suggest the effects of ion beam or soybean 100 random primers were used in the RAPD
DNA on leaf protein of tomato mutants were various, amplification, but only bands amplified by S11 primer
which was same to other researchers (Ji et al., 2001). (GTAGACCCGT) and S45 primer (TGAGCGGACA)
Owing to the effects of ion beam on chromosome could be variant between the control and tomato mutants,
structure (Huang et al., 1994), we infer that variation of and numbers of amplification bands and length of
protein pattern in the leaves of tomato mutants might be amplification fragment were different in the mutants by
caused by the changes of genomic DNA due to the different primer (Figure 3). As shown in the Figure 3 (a),
damage of ion beam or integration of soybean DNA. only one DNA fragment with 550 bp was amplified by
RAPD analysis of genomic DNA in tomato mutants primer S11 in the control, mutant 3 and mutant 11.
RAPD technology is actually PCR amplification, Compared with the control, DNA fragment with 850 bp
and any organism could be identified by RAPD markers increased in mutant 2, mutant 4-8, mutant 10 and mutant
(Williams et al., 1990, Welsh et al., 1991). Hither to, 13, DNA fragment with 550 bp lost in mutant 9, mutant
some plant mutants induced by ion beam implantation 12, mutant 14 and mutant 15, and numbers of
have been already analyzed by RAPD markers, such as amplification bands and length of amplification fragment
Nicotiana tabacum (Zhang et al., 1998), Cucumis melo were same in mutant 12 and mutant 14. Furthermore,
(Chen et al., 2002), Arabidopsis thaliana (Chang et al., four DNA fragments were amplified in mutant 9, in
2003), Dahlia pinnata Cav. (Yu et al., 2008), Jatropha which DNA fragment with 700 bp was also amplified in
Duan et al., 2014
Figure 2: Protein ideograph in the leaves of tomato mutants

1: the control, 2-11: tomato mutant induced by ion beam and soybean DNA, 12: tomato mutant induced with
2×1017N+/cm2 ion beam, 13: tomato mutant induced with 4×10 17N+/cm2 ion beam, 14: tomato mutant induced
with 2×1017Ar+/cm2 ion beam, 15: tomato mutant induced with 4×10 17Ar+/cm2 ion beam.
mutant 12 and mutant 14. On the other side, bands However, rate of differential bands in the mutant 3 and
amplified by S45 primer were shown in Figure 3 (b); two mutant 11 was 0.0 %, moreover rate of differential bands
bands were amplified from the control, mutant 2-6, in other mutants was in the scope of 20.0-50.0 %. Further
mutant 11 and mutant 13, one special band was more, although rate of differential amplification bands
amplified in mutant 8, mutant 10, mutant 12 and mutant was 100.0 % in mutant 9, some protein bands only
15 compared with the control. Moreover, three bands darken and number of protein bands did not change in
were amplified in mutant 9, but their lengths were mutant 9. In addition, the variation of protein pattern in
different from the control and other mutants. Meanwhile, mutant 12, mutant 14 and mutant 15 were relatively
there were two bands in mutant 14 in which one DNA large, and rate of differential amplification bands was
fragment with 700 bp was also found in mutant 12 and respectively 66.7 %, 83.3 % or 100.0 %. Therefore, the
mutant 15, yet other DNA fragment with 500 bp was differential DNA fragments amplified by RAPD might
only amplified in mutant 14. be related to the expression of some genes by encoding
In addition, RAPD amplification bands of tomato some proteins or regulating protein synthesis, but it is not
mutants by S11 and S45 primer were given in Table 1, clear whether differential DNA fragments could
total number of amplification bands, number of influence fruit quality.
differential bands and rate of differential bands in tomato Fruit quality of tomato mutants
mutants were found to be different. Compared with the As everyone knows, tomato is rich in nutrition,
control, rate of differential bands were 100.0 % in mutant such as saccharide, vitamin C, protein, etc. (Xue et al.,
9 and mutant 15, and number of differential bands were 7 2004, Wang et al., 2010). In this research, fruit quality of
and 3, respectively. Secondly, rate of differential bands tomato mutants were assayed, content of vitamin C,
in mutant 14 and mutant 12 were also high, the number soluble saccharide and protein in the fruit of tomato
of differential bands were 5 and 4, respectively. mutants were respectively listed in Table 2. As compared

Duan et al., 2014
a b
Figure 3: Results of RAPD amplification by S11 primer and S45 primer

(a) Results of RAPD amplification by S11 primer, (b) Results of RAPD amplification by S45 primer. M: DM2000,
M: marker, 1: the control, 2-11: tomato mutant induced by ion beam and soybean DNA, 12: tomato mutant induced
with 2×1017N+/cm2 ion beam, 13: tomato mutant induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced
with 2×1017Ar+/cm2 ion beam, 15: tomato mutant induced with 4×10 17Ar+/cm2 ion beam.
with the control, content of vitamin C in 50 % mutants (174.49 μg g-1), moreover content of vitamin C was the
was low, such as mutant 2-4, mutant 6, mutant 9, mutant highest in mutant 11 (242.24 μg g-1). In addition, content
13 and mutant 15, especially lower in mutant 2, mutant 9 of soluble saccharide in 64 % mutants was lower than the
-1 -1
and mutant 4, and was 66.60 μg g , 69.65 μg g and control, but was high in mutant 2, mutant 5, mutant 7,
-1
74.43 μg g , respectively. However, content of vitamin mutant 9 and mutant 10, particularly higher in mutant 7
C was high in mutant 5, mutant 7, mutant 8, mutant 10- (58.84 mg g-1) and mutant 2 (46.96 mg g-1). Furthermore,
12 and mutant 14, especially was higher in mutant 8 content of protein was high in 64 % mutants in
-1 -1
(152.03 μg g ), mutant 10 (167.09 μg g ) and mutant 12 comparison with the control, especially was the highest
Table 1: RAPD amplification bands of tomato mutants by

S11 and S45 primer
Total number Number of Rate of differential

Plants
of bands differential bands bands (%)
1 3 0 0.0
2 4 1 25.0
3 3 0 0.0
4 4 1 25.0
5 5 2 40.0
6 5 2 40.0
7 6 3 50.0
8 4 2 50.0
9 7 7 100.0
10 5 2 40.0
11 3 0 0.0
12 6 4 66.7
13 5 1 20.0
14 6 5 83.3
15 3 3 100.0
1: the control, 2-11: tomato mutant induced by ion beam and soybean
DNA, 12: tomato mutant induced with 2×10 17N+/cm2 ion beam, 13: tomato
mutant induced with 4×1017N+/cm2 ion beam, 14: tomato mutant induced
with 2×1017Ar+/cm2 ion beam, 15: tomato mutant induced with 4×1017Ar+/
cm2 ion beam.

Duan et al., 2014
Table 2: Content of vitamin C, soluble saccharide and protein in the fruit of tomato
Content of vitamin C Content of soluble Content of protein

Plant
(μg/g) saccharide (mg/g) (mg/g)
1 111.95 19.18 18.88
2 66.60 46.96 13.98
3 95.07 13.17 20.51
4 74.43 17.09 26.44
5 114.28 21.37 18.48
6 95.66 14.19 20.12
7 116.91 58.84 29.19
8 152.03 16.35 17.45
9 69.65 37.48 46.57
10 167.09 40.51 6.17
11 242.24 11.06 18.58
12 174.49 16.46 25.48
13 92.36 13.65 21.19
14 122.95 19.12 24.86
15 99.96 12.62 20.35
The average content
119.71 23.87 21.88
in mutants
1: the control, 2-11: tomato mutant induced by ion beam and soybean DNA, 12: tomato
mutant induced with 2×1017N+/cm2 ion beam, 13: tomato mutant induced with 4×1017N+/cm2
ion beam, 14: tomato mutant induced with 2×10 17Ar+/cm2 ion beam, 15: tomato mutant
induced with 4×1017Ar+/cm2 ion beam.
in mutant 9 (46.57 mg g-1), yet content of protein in quality of mutant 3 and mutant 11 are obviously different
mutant 2, mutant 5, mutant 8, mutant 10 and mutant 11 with the control, but rate of differential amplification
was lower than the control, and only 6.17 mg g-1 protein bands was 0.0 % in mutant 3 and 11 which were treated
in mutant 10. with ion beam and soybean DNA, inferring some big
On the other side, content of vitamin C, soluble insert segment of soybean DNA might be not amplified,
saccharide and protein were different in mutants, and perhaps there might be a more complicated relationship
fruit quality of mutants was multifarious. As shown in between nutritional quality of fruit and genomic DNA of
Table 2, compared with the control, content of vitamin tomato irradiated with ion beam or treated with ion beam
C, soluble saccharide and protein in mutant 7 was all and soybean DNA, moreover the effect mechanism of
higher, so mutant 7 has better comprehensive quality of ion beam or foreign DNA was very complex and need to
fruit, secondly were mutant 12 and mutant 14 because be further studied and explored.
content of vitamin C and protein was both higher.
Moreover, content of soluble saccharide and protein in CONCLUSION
mutant 9 was both higher, especially content of protein This study shows that ion beam or soybean DNA
-1
was the highest (46.57 mg g ). However content of could influence leaf protein, genomic DNA and fruit
vitamin C in mutant 11 was the highest (242.24 μg g-1), quality of tomato mutants, inferring the variation of leaf
and content of soluble saccharide and protein was only protein and fruit quality in tomato mutants might be
-1 -1
11.06 mg g and 18.58 mg g . In addition, content of caused by the changes of genomic DNA which would
vitamin C and soluble saccharide was low in mutant 15 happen due to damage of ion beam or integration of
and mutant 3, one other thing to note is that nutritional soybean DNA. However the effects of ion beam or

Duan et al., 2014
soybean DNA were different, and the changes among 6(5): 355-358.
protein, DNA and fruit quality was not consistent with
Duan HY, Yu YA, Li XW and Duan ZQ. 2012.
each other, thus it is necessary to further study effect
Summary of plant transformation mediated by low
mechanism of ion beam or foreign DNA, which would
energy ion beam. Biology Teaching. 37(1): 12-13.
contribute to provide foundation for ion beam mutation
breeding of tomato. Gao WJ, Su JX, Xie L, Deng CL, Zhang T and Lu
LD. 2012. The point mutation induced by the low-energy
ACKNOWLEDGMENT N+ ion implantation in impatiens balsamine genome.
This research was kindly supported by Science Russian Journal of Genetics. 48(10): 1009-1014.
Fund from Henan province (122300410025), and the
Huang WC, Fan SJ, Huang JN, Sang JL, Shi YF, Xia
grant of young teachers in Henan province institution of
ZE, Wu WJ and Yao HL. 1994. Study on mutagenic
higher learning (2011GGJS-063), in P. R. China.
effect of ion implantation into microorganism. Journal of
Anhui Agricultural University. 21(3): 282- 285.
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Journal of Research in Biology ISSN No Print: 2231 –6280; Online: 2231- 6299
Original Research
The leaping behavior of the sally lightfoot crab Grapsus grapsus

(Crustacea: Decapoda: Brachyura) at an oceanic archipelago
Authors: ABSTRACT:
Marina de Sá Leitão
Câmara de Araújo. The genus Grapsus includes a total of nine recognized species of semi-
terrestrial crabs. Among them, Grapsus grapsus (Linnaeus, 1758) stands popularly
Institution: known as sally lightfoot crab. It is very abundant in Oceanic Islands, such as the
Departamento de Ciências Fernando de Noronha Archipelago, Brazil. The present study registered the behavior
Exatas e Naturais, Faculdade of jumping between the rocks by G. grapsus in the supralittoral of Fernando de
de Ciência, Educação e Noronha Archipelago. Field observations were performed in May 2012, including
Tecnologia de Garanhuns video footage. The crabs, juveniles and adults, males and females, jump from a rock to
(FACETEG), Campus another. This can be related to a defense habit, but it seems that the crabs also jump
Garanhuns, Universidade de to avoid entering into the sea, or to escape from wave wash. Other registers on crabs
Pernambuco (UPE), Brazil. jumping from literature are also discussed. However, more studies on this behavior
are still necessary for understanding them completely.

Marina de Sá Leitão Crab behavior, Fernando de Noronha Archipelago, Red rock crab,
Câmara de Araújo. Semi-terrestrial crab.
Article Citation:
Email Id:
Marina de Sá Leitão Câmara de Araújo.
The leaping behavior of the sally lightfoot crab Grapsus grapsus (Crustacea: Decapoda:
Brachyura) at an oceanic archipelago.
Dates:
Web Address:
http://jresearchbiology.com/ Received: 20 May 2014 Accepted: 30 May 2014 Published: 26 Jun 2014
documents/RA0452.pdf.
1357-1364 | JRB | 2014 | Vol 4 | No 4

An International
Araújo, 2014
INTRODUCTION and rocks, which is found under jurisdiction of the State

The genus Grapsus Lamarck, 1801 (Grapsidae) of Pernambuco, Northeast of Brazil. The benthic fauna
includes a total of nine recognized species of semi- of FNA was studied by Lopes and Alvarenga (1955) and
terrestrial crabs: G. adscensionis (Osbeck, 1765), Matthews and Kempf (1970) (Mollusca), Pires et al.,
G. albolineatus Latreille, in Milbert, 1812, (1992) (Cnidaria), Mothes and Bastian (1993) and
G. fourmanoiri Crosnier, 1965, G. granulosus H. Milne Muricy and Moraes (1998) (Porifera), among others.
Edwards, 1853, G. grapsus (Linnaeus, 1758), Several oceanographic expeditions explored the
G. huzardi Desmarest, 1825, G. intermedius de Man, archipelago, such as H.M.S. Beagle Challenger
1888, G. longitarsis Dana, 1851 and Expedition, Hartt Expedition, Branner-Agassiz
G. tenuicrustatus (Herbst, 1783) (WORMS, 2013; Ng Expedition, Calypso, Canopus and Almirante Saldanha.
et al., 2008). Among these species, G. grapsus, stands The results of the Crustacea sampled on these
out popularly and are known as red rock crab, sally expeditions can be found at several publications, such as
lightfoot crab, "aratu" (in Portuguese) and "abuete negro" Smith (1869), Miers (1886), Henderson (1888), Bate
or "sayapa" (in Spanish). This species is found in the (1888), Rathbun (1900, 1918, 1925, among others),
Pacific Ocean, from Baja California to Northern Chile, Forest and de Saint-Laurent (1967) and Coelho et al.,
and Galapagos Islands, and in the Atlantic Ocean, from (2006, 2007, 2008). Fausto-Filho (1974) presented a list
Bermudas, Florida, Gulf of Mexico, Antilles, Colombia, of the Decapoda and Stomatopoda collected by himself
and from Venezuela to Brazil. In the Brazilian coast, this and based on some of the cited publications, which
crab is found from the States of Ceará to Espírito Santo, resulted in a total of 66 species (3 Stomatopoda and 63
but it is more abundant in the Oceanic islands (Fernando Decapoda) for FNA. Included, there is G. grapsus. The
de Noronha Archipelago, Rocas Atoll and Saint Peter species was considered very abundant, being found in all
and Saint Paul Rocks) (Melo, 1996; Freire et al., 2011). beaches. There is no doubt that the species inhabiting
At Saint Peter and Saint Paul Rocks, (Ross 1847, apud FNA is G. grapsus. They are commonly observed in the
Holthuis et al., 1980) cited that this species is a predator rocky shores of the islands that compose the archipelago,
of the eggs of birds that nest at the area, and Viana et al., sharing the habitat with Plagusia depressa (Fabricius,
(2004) cited that this is one of the most abundant animal 1775) (Plagusiidae). The present study aims to describe
species on the rocks. Melo (1996) also signals the the jumping behavior of Grapsus grapsus at FNA during
occurrence of this species at Trindade, a Brazilian field observations.
volcanic island distant 1,167 km from the continent, but
probably the species inhabiting this island is, in fact, MATERIAL AND METHODS
G. adscensionis (Hartnoll, 2009). Ratti (2004) believed The archipelago is distant 545 km from the
that the differences between G. adscensionis and capital of Pernambuco, the Municipality of Recife,
G. grapsus were not enough to support two different occupies an area of 26 km² and the main island,
species, but more recently, several authors such as Ng Fernando de Noronha, has an area of 17 km², being 6
et al., (2008) and Freire et al., (2011), recognized the miles long and 2 miles wide (Matthews and Kempf,
taxonomic validity of both species. 1970; Fausto-Filho, 1974). In May 2012, during three
Among the oceanic island this species can be days, field observations and footages of this species were
found, stands out the Fernando de Noronha Archipelago performed at Sueste Bay, FNA (Figure-1) (3º52'01" S;
(FNA) (3°51′S, 32°25′ W), a complex of volcanic islands 32º25'19" W). At the bay, the Sueste Beach and the
Araújo, 2014
A B
C D
Figure 1. Brazilian coast with the location of the Fernando de Noronha Archipelago, FNA
(A); FNA with the location of the Sueste Bay (B); Aerial view of the Sueste Bay (C);
Rocky shore at Sueste Bay, where the field observations of Grapsus grapsus (Linnaeus,
1758) were perfomed (D).
Sueste Mangrove are included, the last one being behavior of crabs in the literature.
considered the only oceanic mangrove of South Atlantic. The air temperature and tidal heights for the
In the seawater of the bay, there are several islets, such dates of study were obtained through the Integrated
as Cabeluda, Chapéu, Ovos and Trinta-Réis. System of Environmental Data (SINDA).
The individuals of Grapsus grapsus were
observed in the rocky shore of the bay. These rocks are RESULTS AND DISCUSSION
mainly distributed in the extremities of the bay, and also The air temperature for the study period varied
serve as habitat for Plagusia depressa. The water was from 25.5 to 30ºC (Figure-2), characterizing a tropical
transparent and shallow, with a depth of 1m. The footage climate. The observations were performed during the dry
was performed with a Panasonic camera, DMC-FT10 period, equatorial summer. According to Ribeiro et al.,
model. After that, a bibliographic research was (2003, 2005), the FNA climate is of the type Aw of
performed to seek possible registers of the jumping Köppen's classifications, i.e. tropical with semi-arid

Araújo, 2014
Figure 2. Air temperature by dates and hour during the study period,
at Fernando de Noronha Archipelago.
characteristics, having well defined dry and rainy The observed population consisted of Grapsus
periods. grapsus juveniles and adults of both sexes. They were
The tidal level for the study period varied from found sharing the habitat with Plagusia depressa.
1.25 to 2.75 m (Figure-3). The tidal regime can be Besides the size, adults and juveniles are also
characterized as semi-diurnal tide, since there are two distinguished by the color of the carapace. Juveniles of
high tides in each lunar day (Thurman, 1997). According G. grapsus are dark green, dark gray or almost black,
to Souza (2011), the maximum height of the tide in FNA which is important for they camouflage on the
is 2.80 m, and the minimum, 0.0m. Thus, regarding its black volcanic rocks of oceanic islands, and with light
amplitude, the tide of FNA can be classified as yellow spots. On the other hand, adults are quite variable
mesotides. in color; some are dark red or bright red (especially
Figure 3. Tidal level by dates and hour during the study period,
at Fernando de Noronha Archipelago.

Araújo, 2014
Some other interesting information was found in

the literature, regarding the locomotion of crabs. The
species Armases roberti (H. Milne Edwards, 1853)
(Sesarmidae) is found along river banks between rocks
and stones, as well as on the vegetation (Chace and
Hobbs, 1969). According to Schubart and Diesel (1998),
when these crabs are disturbed, they jump from the trees
into the water, and due to this behavior, they are know in
the Caribbean as “jumpy crabs”. Thus, this behavior
could be related to a defensive attitude. A similar
behavior was also registered for Percnon gibbesi (H.
Milne-Edwards, 1853) (Percnidae) by Deudero et al.,
Figure 4. Crabs of the species Grapsus grapsus
(Linnaeus, 1758) from the rocky shore at Sueste (2005); the specimens, observed in shallow waters, run
Bay, Fernando de Noronha Archipelago.
and jump when threatened, seeking for shelter from
males), others are dark green. Some lines and spots can predators.
be observed (Fausto-Filho, 1974; Freire et al., 2011) The crabs Sesarma trapezoideum H. Milne
(Figure-4). Edwards, 1837 (Sesarmidae) occur preferentially in
During the field observations, an unusual riverine cliffs near water streams (Jeng et al., 2003).
behavior in Brachyura could be noticed: the sally According to these authors, these crabs retreat into
lightfoot jumps from a rock to another. Two scenes of crevices or jump into the water below them when
G. grapsus jumping were recorded (Videos 1, 2 and 3). disturbed; few minutes after that, they climb back to the
This behavior was observed for both males and cliff. The species Leptograpsus variegatus (Fabricius,
females, and juveniles and adults. A total of 12 1793) (Grapsidae), a supralittoral crab of rocky shores as
observations were performed. In a first moment, it can be G. grapsus, jump into tidal pools or into the sea to escape
an useful strategy to prevent predation, as described to from predation (Greenaway et al., 1992).
the species which will be discussed below. Besides, this
type of movement could be important to escape from the CONCLUSIONS
wave wash (Video 1) or to avoid entering into the water All these mechanisms described in literature are
(Video 2), instead of walking through the water to reach related to a fast escape from danger, such as predation,
another point of the rocks. They also seem to jump from including jumping into the water. But during the field
a lower to a higher rock (Video 3). Kramer (1967) also observations of G. grapsus, it could be noticed that the
observed this behavior in a population of G. grapsus specimens also jump from a rock to another, which could
from Galapagos. He noticed that the jumpy crabs had an be useful to escape from the wave wash or to avoid
average carapace width of 30 cm. The crabs from FNA entering into the water. They also seem to jump to a
were not measured, but it was clear that they did not higher rock. However, further studies on this feature are
reach 10 cm CW. Before jumping, the crab aligns the still necessary. For example, to test if there is difference
body by stretching the front running pairs of legs on in the jumping frequency between sexes and age classes,
(Kramer, 1967), which was also noticed in the present as well as or to correlate the distance or amplitude of the
study. jumps with the body size of the crab.

Araújo, 2014
ACKNOWLEDGEMENT waters. Mar. Ecol. Prog. Ser., 285: 151-156.

The author is thankful to Maurício de Sá Leitão
Fausto-Filho J. 1974. Stomatopod and decapod
Dévé, Silvia de Sá Leitão Dévé e Jean Luc Dévé for
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Serial No Accession No Title of the article Page No

2 RA0447 Daily Activity Budget of Nicobar Long-tailed Macaque (Macaca 1338-1347

An International Scientific Research Journal

Laboratory evaluation and comparative study of herbal mosquito coils

Susheela P* and Radha R.

Corresponding author: Keywords:

This article is governed by the Creative Commons Attribution License (http://creativecommons.org/

1332-1337 | JRB | 2014 | Vol 4 | No 4

INTRODUCTION contain active materials that are potential enough to

Concentration of lemon grass oil

Concentration of eucalyptus oil

Journal of Research in Biology (2014) 4(4): 1332-1337 1334

Concentration of thyme oil

Figure-3 Repellency of thyme oil against C. quinquefasciatus

RESULTS AND DISCUSSION A number of studies have been focused on lemon

Kitzmiller JB and Micks DW. 1954. Techniques for

Journal of Research in Biology (2014) 4(4): 1332-1337 1336

edible plant-derived essential oils against the pyrethroid-

Zhu J, Zeng X, Ma Y, Liu T, Ting Liu Y, Qian K,

Submit your articles online at www.jresearchbiology.com

1337 Journal of Research in Biology (2014) 4(4): 1332-1337

An International Scientific Research Journal

Daily Activity Budget of Nicobar Long-tailed Macaque

Corresponding author: Keywords:

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1338-1347 | JRB | 2014 | Vol 4 | No 4

INTRODUCTION Veira, 2002; Hamada et al., 2008). Previous researches

Fig 1. Study area and Study site.

1339 Journal of Research in Biology (2014) 4(4): 1338-1347

Journal of Research in Biology (2014) 4(4): 1338-1347 1340

Activity Observation Percentage (%) Expected frequency χ2 = (O-E)2/E

Feeding 321 22.35 179.5 111.54*

Resting 226 15.74 179.5 12.04*

Grooming 160 11.14 179.5 2.12*

Playing 81 5.64 179.5 54.05*

Sexual 21 1.46 179.5 139.95*

Agonistic 08 0.56 179.5 163.85*

Total 1436 100 1436 1156.25

* Showing significant differences (p<0.05), by using the chi-square test (χ2).

1341 Journal of Research in Biology (2014) 4(4): 1338-1347

Fig 2. Various activities of Long-tailed Macaque in Great Nicobar Island

Journal of Research in Biology (2014) 4(4): 1338-1347 1342

Fig 3. Disability in Nicobar Long-tailed macaque

Journal of Research in Biology (2014) 4(4): 1338-1347 1344

Kipper S and Todt D. 2002. The use of vocal signals in

1345 Journal of Research in Biology (2014) 4(4): 1338-1347

Journal of Research in Biology (2014) 4(4): 1338-1347 1346

Umapathy G, Mewa Singh and Mohnot SM. 2003.

Van Noordwijk MA and van Schaik CP. 1999. The

Wheatley BP. 1980. Feeding and Ranging of East

Zamzarina MA. 2003. A study on aggressive