Wei ZHANG, Ben LI, Xiaoxiao SHU, Hanbin XIE, Enle PEI, Xiao YUAN, Yujie SUN, Tianhou WANG*and Zhenghuan WANG*

1School of Life Science, East China Normal University, Shanghai200062, China

2Shanghai Landscaping and City Appearance Administrative Bureau, Shanghai200040, China

A New Record of Kaloula （Amphibia: Anura: Microhylidae） in Shanghai， China

Wei ZHANG1, Ben LI1, Xiaoxiao SHU1, Hanbin XIE1, Enle PEI2, Xiao YUAN2, Yujie SUN2, Tianhou WANG1*and Zhenghuan WANG1*

1School of Life Science, East China Normal University, Shanghai200062, China

2Shanghai Landscaping and City Appearance Administrative Bureau, Shanghai200040, China

We discovered a medium-sized species of narrow-mouthed frog of the genus Kaloula in June and September 2014 during municipal surveys of amphibians in Shanghai. Three narrow-mouthed frogs were collected in city drains of the Binjiang Forest Park, Shanghai. Based on subsequent examination and morphological characters, these specimens were identifi ed as Kaloula borealis (Barbour, 1908). The species was mainly distributed in Northern China and Korea. It is a new amphibian record for Shanghai. We found the visible differences in male secondary sexual characteristics between our specimens and Beijing specimens. K. borealis was fi rst described inhabiting urban greenlands which could benefi t amphibian conservation in urban and urbanizing areas.

Kaloula borealis, Urban greenland, Amphibian, new record

The genus Kaloula Gray, 1831 (Anura: Microhylidae) is currently composed of 17 recognized species and is distributed throughout Northern China, Korea, Lesser Sunda Islands, Philippines, Bangladesh, India, and Sri Lanka (Diesmos et al., 2002; Das et al., 2004; Fei et al., 2009; Frost, 2014). This genus is composed of only fi ve species in China (Mo et al., 2013). However, no species of this genus has been reported in the area of Shanghai Municipality. A 3 year wildlife survey in Shanghai Municipality was initiated in June 2013. We found 8 narrow-mouthed frogs (Kaloula borealis) after a rainstorm in city drains of Binjiang Forest Park on 22 June 2014 (Site A: 31°23′1″ N, 121°31′20″ E) (Figure 1). We later counted 51 individuals around a pond of the same park on 4 September 2014, and another 23 individuals in greenlands near a small stream of Wusong Coastal Forest Park next day (Site B: 31°23′38″ N, 121°30′23″ E). Thisspecies was fi rst described as inhabiting urban greenlands; two females and one male were collected, euthanized, and fi xed in formalin, then transferred to 70% ethanol. Muscle or liver tissues were sampled and preserved in 100% ethanol for DNA extraction. All specimens (Voucher Nos.: ECNU14062203; ECNU14062204; ECNU14062205) were archived at the School of Life Science of East China Normal University, China.

Body measurements: SVL (snout–vent length: from the tip of the snout to the vent); HL (head length: distance between jaw angle and snout tip); HW (head width: across jaw angle); SL (snout length: from anterior eye front to snout tip); IND (internarial distance); IOD (interorbital distance); UEW (maximum upper eyelid width); ED (eye diameter); LAHL (length from the elbow to the tip of the third fi nger); HLL (hind leg length: from the vent to the tip of the longest toe); TL (tibia length); TW (tibia width); FOL (foot length: from the base of the inner metatarsal tubercle to the tip of the fourth toe). We obtained comparative morphological data from previous studies on Kaloula specimens and from photographs of live specimens (Table 1). We used Independent T-tests to compare our measuring data with previous data. Our results indicate that these morphological parameters of our specimens are consisted with K. borealis(ECNU14062203: Levene’s test F = 0.038, P= 0.847, T-test P= 0.946; ECNU14062204: Levene’s test F = 0.001, P = 0.972, T-test P= 0.984; ECNU14062205: Levene’s test F= 0.128, P= 0.724, T-test P= 0.757).

Table 1 Morphological comparison of Kaloula borealis collectiong from Beijng and Shanghai (in mm).

We assigned this specimen to K. borealis according to the following characteristics: medium size (41.7–42.9 mm in two adult females, 37.3 mm in one adult male), smooth or slightly rough olive dorsum with irregular dark-green marks and brown spots (Figure 2a), half-webbed toes (except for the fourth toe), tips of both fingers and toes not dilated, obtuse upper surface of the tips of both fi ngers and toes (Figure 2d), and sides with small lemon-colored spots (Figure 2b). However, these specimens differed from those described by Fei et al. (2009) in secondary sexual characteristics. The specimens from Shanghai had fl esh-colored throats, no epidermal adhesive glands in the chest and belly, thin and semitransparent ventral skin, and white linea masculina (Figure 2c).

Kaloula borealis was found in Binjiang Forest Park and Wusong Coastal Forest Park, which are adjacent to the Changjiang River and to the banks of the Huangpu River, respectively (Figure 3). Local climate is subtropical monsoon, with an annual mean temperature of 16.9oC and average annual rainfall of 1213 mm. These two parks were reclaimed from the Changjiang River in the 1950s and the 1960s. The landscape is mainly secondary forest, with fresh-water ponds connected by a relatively dense short river network. The vegetation is composed of an artifi cially planted forest predominated by trees, such as camphor tree (Cinnamomum camphora), bald cypress (Taxodium distichum), pond cypress (T. ascendens) and southern magnolia (Magnolia grandiflora), the aquatic plants around the wetlands were common reed (Phragmites australis), wall iris (Irissp.), great bulrush (Schoenoplectus sp.), and shrubs (Fatsia japonica, Ligustrum quihoui, Pittosporum tobira, Viburnum odoratissinum). Elevation of sampling sites ranged from 5 to 15 m, with a temperature of 25oC and humidity of 95% as we measured in the field. The habitats of the frog adjoined the wetlands of the park (Figure 4), and K. borealis gathered in the drains for breeding after rainstorm.

Discussion

We discovered a medium-sized species of narrowmouthed frog (K. borealis) of the genus Kaloula in June and September 2014 during municipal surveys of amphibians in Shanghai. This is a new record of the genus Kaloula and K. borealis in Shanghai. The morphological data and characteristics of K. borealis from Shanghai were consistent with Beijing specimens except secondary sexual characteristics. Shanghai specimens have fleshcolored throats, no epidermal adhesive glands in the chest and belly, thin and semitransparent ventral skin, and white linea masculina (Figure 2c). By contrast, Fei et al. (2009) described the specimens of Beijing with dark gray throats, epidermal adhesive glands in the chest but notin the belly, and mulberry-colored masculina. Due to the limited sampling size, we cannot conclude whether the differences between the two populations are significant or not. Molecular phylogeny research will be performed in near future, and identify the difference of secondary sexual characteristics (Matsui et al., 2011).

Our field surveys indicated that K. borealis has inhabited the urban greenlands adjoining the mouth of the Huangpu River. In addition, we observed reproductive activity and eggs of K. borealis in a pond in Binjiang Forest Park, and this species has formed a population in artificial ponds and stormwater wetlands of urban greenlands. Upon communicating with local ecologists, we learned that one college student found individuals of this species in Fudan University’s campus in Jiangwan Town (Site C: 31°20′24″N, 121°30′16″E) in 2008, which is 5 km from Site A. This account was further confi rmed by the website of China with K. borealis’s picture (Internet reference, 2012). Considering the absence of K. borealis during a herpetological survey of Shanghai in the 1990s (Shanghai Forestry Bureau, 2000), we speculated that this species has only migrated to the urban greenlands of Shanghai in recent years. Previous studies on the herpetology of Shanghai from the 1960s to present indicated that our present work is the first record on K. borealis in Shanghai, despite this species being distributed in adjacent provinces, such as Zhejiang and Jiangsu Province (Fei et al., 2009). We also discovered evidence of this frog using artifi cial greenlands for breeding along the Huangpu River in Shanghai. The number of known amphibian species was up to 13 in Shanghai, including Bufo gargarizans, B. raddei, Hyla immaculata, H. chinensis, Hylarana guentheri, Fejervarya multistriata, Pelophylax nigromaculatus, Pelophylax plancyi, R. japonica, Hoplobatrachus chinensis, Microhyla ornata, Rhacophorus dennysi, and Polypedates megacephalus, according to the records of 1980 (Huang et al., 1980). However, only 8 species of frogs had been found in the first wildlife survey of Shanghai Municipality in 2000 (Shanghai Forestry Bureau, 2000), with B. raddei, H. chinensis, H. guentheri, R. dennysi, and P. megacephalus still missing during surveys. From 2013 to 2014, thesecond wildlife survey of Shanghai Municipality was undertaken. During these surveys, we counted 5 species frogs (B. gargarizans, F. multistriata, P. nigromaculatus, P. plancyi M. ornata) and 1 new record species (K. borealis). By contrast with the surveys in 2000, we did not encounter H. immaculata, H. chinensis and R. japonica in these surveys.

Urbanization is recognized as a major driver of amphibian declines globally (Hamer et al., 2008). Urban greenlands is useful for mitigating habitat loss and retaining water runoff from impermeable urban surfaces, and serve as breeding habitat for amphibians. Our studies show that urban greenlands play increasingly important roles as an amphibian refuge due to urbanization in the Changjiang Estuary. Amphibians require aquatic habitats to support larval development and terrestrial habitat for adult stages. Urban greenlands provide necessary habitat for many species that spend breeding season in ponds, but forage and hibernate in the surrounding riparian zone and upland vegetation (Scheffers and Paszkowski, 2013). In areas with high levels of human activity, urban gardens and greenland parks are likely the last available habitats for amphibians and have tremendous value in preserving the diversity of amphibian species. Moreover, K. borealis is distributed along the two banks of the Huangpu River. We hypothesize that the river plays a key role in dispersal, and most researchers agree that rivers help create and sustain species diversity (Lougheed et al., 1999; Li et al., 2009). With further study and extensive field work, we speculate that K. borealis may be found in more localities, which will be useful to understand the exact distributional pattern of K. borealis in Shanghai.

Acknowledgements We thank Qifeng ZHU from Binjiang Forest Park, Xiaokang Yin from Wusong Coastal Wetland Forest Park, and Dr. Yuyi LIU from Shanghai Wild Plant and Animal Protection, Department for Issuance of a Research Permit. This work was supported by the Shanghai Landscaping and City Appearance Administrative Bureau Project (Grant No. F131508).

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s: Zhenghuan WANG, from East China Normal University (ECNU), Shanghai, China, with his research focusing on urban ecology and wildlife biology; Prof. Tianhou WANG, from ECNU, Shanghai, China, with his research focusing on ecology and conservation biology.

E-mail: zhwang@bio.ecnu.edu.cn (Z. H. WANG); thwang@bio.ecnu. edu.cn (T. H. WANG)

17 October 2014 Accepted: 16 April 2015