© 2005 The Japan Mendel Society Cytologia 70(3): 249–255, 2005 Cytogenetic Study on Wild Animal Species of the Subfamily Paradoxurinae in Thailand Alongkod Tanomtong1, Arunrat Chaveerach1,*, Apiradee Sriphoom2 and Ruengwit Bunjonrat2 1 Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen, 40002, Thailand 2 Genetics Program, Department of Botany, Faculty of Science, Chulalongkorn University, Payathai, Bangkok 10300, Thailand Received April 11, 2005; accepted May 18, 2005 Summary As an endangered species in Thailand, wild animal species of the subfamily Paradoxuri- nae were selected for a cytogenetic study. The animal species sampled are Masked palm civet (Pagu- ma lavata), Binturong (Arctictis binturong), Asian palm civet (Paradoxurus hermaphroditus) and Small-toothed palm civet (Arctogalidia trivergata). Blood samples taken from Khao Khiew Zoo, Songkhla Zoo and Dusit Zoo were examined using lymphocyte culture techniques. The results indi- cate that the number of diploid chromosome of Masked palm civet, Binturong, Asian palm civet and Small-toothed palm civet are 44, 42, 42 and 40, respectively. The fundamental number (NF) are 62, 46, and 50 for both male and female Masked palm civet, Binturong, Asian palm civet, while that of Small-toothed palm civet is 56 for female and 55 for male. The type number of autosomes, metacen- tric, submetacentric, acrocentric and telocentric chromosome are 6-10-10-16, 2-4-16-18, 2-4-20-14, and 6-8-10-14, respectively. Chromosome pairs 17, 19, and 20 are satellite chromosomes. The X and Y chromosomes are large submetacentric and small submetacentric for the Masked palm civet, whereas the Binturong and Asian palm civet have large metacentric and small submetacentric. The Small-toothed palm civet contains large metacentric and small acrocentric chromosome. In addition, conventional staining and G-banding techniques show that there are differences in number and loca- tion of G-banding for each species. Key words Cytogenetic, Paradoxurinae, Wild animal. Thailand is a country with high biodiversity, resulting from its varying ecosystems and forests. It has been estimated that there may be up to 100,000 wildlife species in Thailand as well as many unknown plant species. Unfortunately, rushed economic development and social welfare has led to vast destruction of Thailand’s forest and wild life resources. Many wildlife species have become ex- tinct due to loss of habitat, hunting for food, trade and export. One such example is civets, whose numbers are rapidly deteriorating. Masked palm civet, Binturong, Asian palm civet and Small- toothed palm civet are members of the subfamily Paradoxurinae. Their present rarity causes as well great difficulty in obtaining blood samples for cytogenetic studies. Wuster and Benirschke (1967) studied the cytogenetics of Masked palm civet, Binturong, and Asian palm civet, and their results showed chromosome numbers of 44, 42, and 42, respectively. The karyotypes indicate that Masked palm civet have 22 metacentrics and submetacentrics, 20 acro- centrics and telocentrics, and metacentric and submetacentrics X and Y chromosomes. Binturong show NF of 68 for both male and female and karyotypes of 22 metacentrics and submetacentric, 18 acrocentrics and telocentrics, and medium metacentric and small metacentric X and Y chromo- somes. Asian palm civet were reported to have NF of 66 for both male and female, X and the Y chromosomes as medium metacentric, and the smallest chromosome was undetermined. * Corresponding author, e-mail: [email protected] 250 Alongkod Tanomtong et al. Cytologia 70(3) Wang et al. (1984) reported chromosome numbers for Masked palm civet, Binturong, and Asian palm civet to be 2 nϭ44, 42, and 42, respectively. The karyotype of Masked palm civet shows 24 metacentrics and submetacentrics, 18 acrocentrics, and the X and Y chromosomes are submeta- centric and acrocentric. The NF is 69 and 68 for the male and female. The most recent cytogenetic study on Masked palm civet was conducted by Masashi and Haru- mi (1993). Their results showed chromosome numbers of 44. The karyotype showed 8 metacentrics and submetacentrics, 18 acrocentrics and 16 telocentrics, and the X and Y chromosomes are meta- centrics. The NF of both male and female is 66. Studies of chromosomal features in Indian Muntjacs (Muntiacus muntjak) and Fea’s muntjacs (M. feae) of Thailand (Tanomtong et al. 2005) indicate chromosome variation with breakage, fusion and lacking of homologous chromosome in the same species and between sexes. These factors cause abnormalities in productive gametes and may have contributed to the Fea’s muntjacs becom- ing an endangered species. As an endangered species, basic data resulting from cytogenetic studies can help to identify in- nate features which may lead towards the extinction of civets. Therefore, this study aims to test the normality or abnormality of chromosomal features in Masked palm civet, Binturong, Asian palm civet and Small-toothed palm civet. Materials and methods Blood samples of 3 male and 2 female Masked palm civet, 2 male and 2 female Binturong, 1 male and 2 female Asian palm civet, and 2 male and 2 female Small-toothed palm civet from Khao Khiew Zoo, Cholburi province, Songkla Zoo, Songkla province and Dusit Zoo, Bangkok, Thailand were collected and subjected to cytogenetic studies by lymphocyte culture of whole blood samples. The cultured cells were examined with a colchicine-hypotonicfixation-air-drying technique fol- Fig. 1. Metaphase chromosome of male (A) and female Fig. 2. Metaphase chromosome of male (A) and female (B) Masked palm civet (Paguma lavata) 2 nϭ44, (B) Asian palm civet (Paradoxurus hermaphrodi- male (C) and female (D) Binturong (Arctictis tus) 2 nϭ42, male (C) and female (D) Small- binturong) 2 nϭ42, and satellite chromosomes toothed palm civet (Arctogalidia trivergata) (arrows). 2 nϭ40, and satellite chromosomes (arrows). 2005 Cytogenetic Study on Wild Animal Species of the Subfamily Paradoxurinae in Thailand 251 lowed by conventional staining and G-banding with Giemsa’s. Chromosomal checks using light mi- croscopy were performed with 20 cells of each individual. Results This is the first study to describe the chromosome and karyotype of these wildlife animals of Thailand. Chromosome numbers of Masked palm civet, Binturong, Asian palm civet indicate 2 nϭ44, 42 and 42, respectively (Figs. 1, 2). The 44 chromosomes of Masked palm civet consists of 6 metacentrics, 10 submetacentrics, 10 acrocentrics and 16 telocentrics; the sex chromosomes are large submetacentric and small submetacentric; the 17th chromosome pair is a satellite chromo- some (Figs. 3A, B and 5A, B). The 42 chromosomes of Binturong is comprised of 2 metacentrics, 4 submetacentrics, 16 acrocentrics and 18 telocentrics; the X and Y chromosomes are large meta- centrics and small submetacentrics; the 19th chromosome pair is a satellite chromosome (Figs. 3C, D and 5C, D). The Asian palm civet has 42 chromosomes, consisting of 2 metacentrics, 4 submeta- centrics, 20 acrocentrics and 14 telocentrics; the sex chromosomes are large metacentric and small submetacentric; the 20th chromosome pair is a satellite chromosome (Figs. 4A,B and 6A, B). For the first cytogenetic study of the Small-toothed palm civet, the chromosome number was reported 2 nϭ40 (Fig. 2C, D), consisting of 6 metacentrics, 8 submetacentrics, 10 acrocentrics, 14 telo- Fig. 3. Karyotype showing different all types of chromo- Fig. 4. Karyotype showing different all types of chromo- somes for male (A) and female (B) Masked palm somes for male (A) and female (B) Asian palm civet (Paguma lavata) 2 nϭ44, male (C) and fe- civet (Paradoxurus hermaphroditus) 2 nϭ42, male (D) Binturong (Arctictis binturong) 2 nϭ42, male (C) and female (D) Small-toothed palm and satellite chromosomes (arrows) by conven- civet (Arctogalidia trivergata) 2 nϭ40 and satel- tional staining method. lite chromosomes (arrows) by conventional stain- ing method. 252 Alongkod Tanomtong et al. Cytologia 70(3) Fig. 6. Karyotype different all types of chromosomes for Fig. 5. Karyotype showing different all types of chro- male (A) and female (B) Asian palm civet (Para- ϭ mosomes for male (A) and female (B) Masked doxurus hermaphroditus) 2 n 42, male (C) and palm civet (Paguma lavata) 2 nϭ44, male (C) female (D) Small-toothed palm civet (Arctoga- ϭ and female (D) Binturong (Arctictis binturong) lidia trivergata) 2 n 40 and satellite chromo- 2 nϭ42, and satellite chromosomes (arrows) by somes (arrows) by G-banding staining method. G-banding staining method. centrics, and X and Y chromosomes are large metacentric and small acrocentric. There is a genetic marker on the 17th chromosome pair called a satellite chromosome (Figs. 4C, D and 6C, D). The results from G-banding staining on autosomes and sex chromosomes of Masked palm civet, Binturong, Asian palm civet and Small-toothed palm civet show 141, 142, 169 and 159 color bands. NF is 62 for male and female Masked palm civets, 46 for male and female Binturongs, 50 for male and female Asian palm civets, and 56 for male and 55 for female Small-toothed palm civets (Figs. 5, 6, 7, 8, 9, 10). The highly varied G-banding patterns present a basis for a summary of their evolution. The total results of this study are shown in Table 1. Discussions The Present results agree with the previous studies by Ray-Chaudhuri et al. (1966), Wurster and Benirschke (1967), Wang et al. (1984); and Masashi and Harumi (1993). The Masked palm civet, Binturong, Asian palm civet and small-toothed palm civet show chromosome number of 44, 42, 40 and 42, respectively. Variations occurring on autosomes and sex chromosomes cause differ- ences of NF in male and female individuals (Figs. 7–10). According to these 4 sampled species of the subfamily Paradoxurinae in Thailand, the varied chromosome number is in the range of 40-44 chromosomes while the NF ranges from 46–56 chromosomes. The sex chromosomes are meta- centrics or large submetacentrics and submetacentrics or small acrocentric, which differs from re- 2005 Cytogenetic Study on Wild Animal Species of the Subfamily Paradoxurinae in Thailand 253 Fig.