The Morphological Characters of the Malayan Pit Viper Calloselasma rhodostoma (Kuhl, 1824): on The Cephalic Scalation and Distribution Status in Indonesia

The examination on variations of morphological characters among 35 specimens of Calloselasma rhodostoma (Kuhl, 1824) from four different populations in Indonesia has been completed in this study. Univariate and multivariate analyzes allowed us to recognize the clustering of four populations through morphological diagnosis. The results of the average body size (Total Length) showed that the largest male is from Kangean Island (579.33 mm), while the largest female is from Java (841.07 mm). Comparison of meristic analysis represented three clusters from Principal Component Analysis (PCA) which is considered to be independent population. Here we also described three types of cephalic scalation variation that called small accessories scales and their distribution in Indonesia.

collected from North Borneo, which deposited in Museum Zoologicum Bogoriense-Bogor, Indonesia (MZB 428).This species inhabits various habitat types indicating that this species is well adapted in lowland forest, plantations, shrubs, and rocky areas [4].

Morphometric and Meristic Characters Measurement
The measurement of morphometric and meristic characters in this study followed Vogel et al. [11], with modification as listed in Table 1.A total of 37 morphological characters, consisted of 19 morphometric and 18 meristic characters, were examined for each specimen.We used a slide-caliper to the nearest 0.05 mm for morphometric measurements, except Snout Vent Length (SVL) and Tail Length (TaL) which was measured to the nearest 1 millimeter.The counting of ventral scales, body scales, and subcaudal scales followed Dowling [23]; the terminal scute of subcaudal was excluded in this counting; while the counting of dorsal scales row followed Vogel et al. [11].
To prevent juvenile specimens be examined, we determined the size of Total Length (TL) as 400 mm for adult specimens.Pattern and coloration of the body were observed from fresh preserved specimens [11].Here, we also examined all specimens for the variation on cephalic scalation identified as small accessories scale and the distribution in Indonesia.The small accessories scale that we identified as scale that touched to internasal, prefrontal, nostril and loreal scale.

Data Analysis
Univariate analysis was conducted.Each sample was classified into three groups recognized as Java, Karimunjawa, and Kangean population as presented in Table 2.For multivariate analysis, morphometric data were classified into male and female, since snakes exhibited sexual dimorphism.Data analysis were conducted in 6 characters with bootstrap 999 for Principle Component Analysis (PCA) using PAST3 program after standardizing all characters.The standardization was done by dividing all characters with SVL and converting into percentage.The converted data was transformed to log 10 values.

RESULT AND DISCUSSION
In this study, univariate statistical analysis consists of mean, standard deviation, minimum and maximum values of Java, Karimunjawa, and Kangean population as presented in Table 2 for all characters examined.The largest and smallest average males by TL character were 579.33 mm and 498.75 mm from Kangean Island and Karimunjawa Island, respectively.Whereas the largest and smallest average females were 841.07 mm and 573 mm from Java and Karimunjawa Island, respectively.
The meristic comparison that consists of the number of ventral, subcaudal, supralabial, infralabial, and dorsal scale, is presented in Table 3.According to the range of ventral scale number in males, Karimunjawa population shows higher range than Java and Kangean population.That result is in contrast to females, which shows that Karimunjawa and Kangean population is in the range of Java population.Meanwhile, according to subcaudals scale number, Kangean population has a higher range than Java and Karimunjawa population, both in male and female specimens.The range of dorsal scales number in female specimens shows that Java population has the highest range than other populations.
Several characters indicate that C. rhodostoma exhibits sex dimorphism (see Table 2), including SVL, HL, HW, DE-Nostril length, SnL, ratio TaL/TL, ratio SnL/ED, number of ventral scales, and subcaudal scales.The ratio of TaL/TL (males x: 498.75-579.33,females x: 550.67-841.07)shows that females have longer size than males [27,28].The ratio of TaL/TL (males: 0.18-0.20,females: 0.11-0.12)strongly shows that sexual dimorphism has occurred in C. rhodostoma.The benefits of sexual dimorphism in species with few or no parental care are related to its adaptive ability.
Generally, larger females would produce larger and higher number of offspring.Larger offspring would have higher survival rates as it is able to store more yolk for their development.In males, an individual with a smaller size will have a positive impact including enhancing their ability to find and court female.In addition, the smaller size refines the chemosensory senses, higher mobility, inconspicuous to the predator, and early maturation [29][30][31][32][33].The tail length differences between males and females are due to the influence of copulatory organs (hemipenes) inside the tail, which is the size of tail length affecting male mating success [28].Notes: ♂ = male specimen, ♀ = female specimen

Principal Component Analysis (PCA)
Factor loadings of the first two components from Principal Component Analysis (PCA) are presented in Table 4. Principal Component Analysis in males and females are shown in  The first and second score factor loadings of PC females explain a moderate total variability which is similar to males.All variables show positive score loadings in the first PC with SVL and HL as the highest and TaL as the lowest.The  The different variation on head of C. rhodostoma may be affected by the snake's adaptation process with its ecological environment.Various habitats resulted in a variety of adaptation processes that lead to morphological differences in C. rhodostoma.The same location with different ecological conditions is also able to result in different ecogenesis processes [34].In this research, several types of accessories scales were found in Java Island.However, the variation represented different geographical region for example Type 2 was found only in West Java, while Type 1 and Type 3 were found in Central Java and East Java.

CONCLUSION
Morphological variation in chepalic scalation, including three types of small accessories scale between population of C. rhodostoma in Java, Karimunjawa, Kangean Island can be used as one of the supporting data to determine the taxonomic status of the species.However, this finding needs to be tested using molecular studies for a further determination on the taxonomic status of these populations.

Figure 1 .
The two first scores of factor loadings of principal component (PC) males explain a moderate total variability.All variables show positive score loadings in the first PC, with TaL and DE-Nostril Length as the most positive loaded, in contrast with ED which is lower.The second PC shows three variables that are positively loaded, and the other three are negatively loaded.The variables of SVL and TL are the most positively loaded, in contrast to the lower load of HL.The PCA results of males' C. rhodostoma are shown in Figure 1.(A), in this graph, plot of the two first PC show three groups: Cluster II, Cluster III, and Cluster I. Based on the locality, Cluster III is separated with Cluster II and I.However, our results show a little overlap among these two clusters (Cluster II and I).The single specimen of OTU 3 (Cluster II) is placed inside of Cluster I.

Figure 1 .
Figure 1.Plot of component 1 (PC1) versus component 2 (PC2) for the principal component analysis of males and females in three populations of C. rhodostoma.(A) Males; (B) Females.

Figure 2 .
Figure 2. Head scalation of specimen of Calloselasma rhodostoma (MZB 470) from Bogor, West Java, showing the position of a small accessory scale directly adjacent to four other scales (as: accessory scale, in: internasal, pf: prefrontal, l: loreal, n: nasal).

199Figure 3 .
Figure 3. Relative Amount of Pro-inflammatory Cytokines in Experimental Mice after 24 Days of Treatment.

Table 1 .
List of Morphological Characters Used in This Study and Their Abbreviation Malayan Pit Viper: Cephalic Scalation & Distribution Status in Indonesia (Kadafi et al)

Table 2 .
Summary of Morphometric Measurements of C. rhodostoma from Three Different Localities.Data are shown as the mean and standard deviation, followed by the range in parentheses.All measurements are shown in mm.

Table 4 .
Factor loading on the two first components extracted from the correlation matrix of 6 characters for males and females of C. rhodostoma