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Home , Manidae, Manis, Pangolin, Phataginus, Sunda pangolin

Received: 27 May 2017 | Revised: 14 September 2017 | Accepted: 17 October 2017 DOI: 10.1002/zoo.21388

RESEARCH ARTICLE

Keeping and breeding the rescued Sunda ( javanica) in captivity

Fuhua Zhang | Jiaming Yu | Shibao Wu | Shaoshan Li | Cuiyun Zou | Qiaoyun Wang | Ruyong Sun

School of Life Science, South Normal University, , P. R. China The Sunda (Manis javanica Desmarest, 1822) is a Critically Endangered species. Given that this species lacks effective policies for in situ conservation and Correspondence Shibao Wu and Shaoshan Li, Professor, prevention of in the wild, ex situ conservation and a captive breeding program College of Life Sciences, South China Normal are urgently needed to save this species from . However, techniques for the University, No.55, Zhongshan Avenue West, Tianhe District, Guangzhou 510631, P. R. maintenance and captive breeding of pangolins have not been well developed. In China. June 2010, we established the Pangolin Research Base for Artificial Rescue and Email: [email protected] (S.W.); [email protected] (S.L.) Conservation Breeding of South China Normal University (PRB-SCNU). To date, a total of 34 cubs have been born at PRB-SCNU, 26 of which were captive Funding information Natural Science Foundation of Guangdong bred, indicating great progress in keeping and breeding the Sunda pangolin. The Province, Grant number: S2013010013356; techniques for maintenance—including housing, transitioning to an artificial diet, Special Fund for Significant Science and Technology in Social Development Field of husbandry, and captive breeding—are described in this paper. The purpose of this Guangdong Province, Grant number: manuscript is to share our successful experiences and techniques in the maintenance, 2011A030100012; National Natural Science Foundation of China, Grant numbers: management, and captive breeding of Sunda pangolins, and to provide guidance and a 31572286, 31670266, 31702029; reference for the captive care and management of Sunda pangolins and other pangolin Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme species. (2012); China Environmental Protection Foundation KEYWORDS captive breeding, diet transition, husbandry, maintenance,

1 | INTRODUCTION Phillips, 1985; Wu, Wang, & Feng, 2005). This species has a low reproductive rate, as each pangolin always gives birth to only one There are eight extant species of pangolins worldwide belonging to the offspring at a time and breeds only once a year (Zhang, Wu, et al., 2015). mammalian Order Pholidota, Family (Corbet & Hill, 1992; In the last few decades, habitat loss and hunting to supply the Gaubert & Antunes 2005). Also called “scaly-,” because their international trade for live pangolins, , scales, and meat has led to body surfaces are covered by overlapping scales, they consume mainly the wild population declining dramatically and an inverted pyramid age and in the wild (Heath & Vanderlip, 1988). Sunda pangolins structure (Challender, Thai, Jones, & May, 2012; Challender et al., 2014; (Manis javanica Desmarest, 1822) are distributed in Southeast Asia, Challender, Harrop, & MacMillan, 2015; Yang et al., 2015). Therefore, including Malaysia, , , , Lao PDR, Thailand, the Sunda pangolin was assessed as Critically Endangered on the Menglian County of Puer City, and Mengla County of Xishuangbanna, International Union for Conservation of (IUCN) Red List of Yunnan Province, China, and other Southeast Asia regions (Challender in 2014 (Challender et al., 2014), and was also listed et al., 2014; Francis, 2008; Gaubert & Antunes, 2005; Payne, Francis, & in Appendix I of the Convention on International Trade in Endangered Species of Wild Fauna and Flora in 2017 (CITES, 2017), which bans

Shibao Wu and Shaoshan Li contributed equally to this work. international trade in wild caught pangolins for commercial purposes.

Zoo Biology. 2017;1–10. wileyonlinelibrary.com/journal/zoo © 2017 Periodicals, Inc. | 1 2 | ZHANG ET AL.

Since current policies appear ineffective for in situ conservation of the enclosures at PRB-SCNU are in a large room. Housing details for and prevention of poaching in the wild, ex situ conservation is urgently Sunda pangolins have been described by Zhang, Wu, et al. (2015). needed for the Sunda pangolin through the rescued individuals from Experience indicates that the best ambient temperature for Sunda the illegal trade into artificial environments to establish a long-term pangolins is 29–32°C. Pangolins display obvious cold stress (body viable artificial population to save this species from extinction. Ex situ rolled tightly into a ball and even shaking) when the ambient conservation protects endangered and plants by removing the temperature is below 20°C, and large temperature fluctuations can species to an artificial environment or other area and is the last also lead to . In addition, the cold, dry climate in winter conservation scheme for some species that are on the verge of (temperature and humidity are typically below 20°C and 60%, extinction. The principle of ex situ conservation is that can be respectively) can lead to dryness or cracking of the of some maintained and bred in an artificial environment. Since 1929, at least individuals (Figure 1). Therefore, the room is equipped with under- 20 institutions have attempted to keep Sunda pangolins in captivity floor heating and an air humidifier to control the temperature and (Clark, Thai, & Phuong, 2008; Challender et al., 2012; Vijayan, Yeong, & humidity of the enclosures and provide the pangolins with a warm and Ling, 2008; Wang et al., 2015; Yang et al., 2007; Yu, Peng, Liao, Jiang, & humid artificial environment without large fluctuations in temperature Ma, 2016); however, there has been no substantial success in the and humidity. The Sunda pangolin is nocturnal, so we also equip each maintenance and captive breeding of this species, mainly because enclosure with a monitor so that we can observe the undisturbed pangolins are highly specialized animals with poor immunity (Choo behavior of captive animals, record and review activity throughout the et al., 2016) that are unable to adapt to environmental changes; there is night, and collect data on this species’ activity pattern. a lack of knowledge of pangolin biology and ecology to address their Space is an essential resource for survival; thus, limited space is captive needs; and there is a lack of mastery of techniques to maintain likely to be a limiting factor on behavior and may be a source of stress in captive pangolins, with no concrete method as a guideline. captive animals (Jiang, 2004). The home range of some pangolin In the past several years, many live Sunda pangolins have been species is known. The ( tricuspis) has a small smuggled into Guangdong Province, China, and some have been home range of only 150–330 m2 (Pages, 1975). The range of the Cape confiscated by the Forest Police. However, these confiscated pangolin ( temminckii) is 0.17–11.07 km2 (Heath & Coulson, pangolins could not be released cursorily into the wild. Because, 1997), and the mean home range of male Sunda pangolins is 0.41 km2 firstly, China is a marginal range for Sunda pangolins, and the trade- (n = 3) (Lim, 2007). However, captive pangolin enclosures in many confiscated individuals with unknown origin likely emanated from institutions are less than 10 m2, with the smallest only 0.5 m2 (Chin Indonesia, Malaysia, Vietnam, etc, it would cause such as genetic et al., 2011; Ogilvie & Bridgwater, 1967; Vijayan et al., 2008; Wilson, pollution of populations if these Sunda pangolins were released 1994; Yang et al., 2007). This suggests that the sizes of captive elsewhere in its range; Secondly, it is very difficult to survival for these enclosures are very different (too small) from the natural home range rescued individuals when they are released into elsewhere in their area, which may be a major reason for the failure of captive pangolins. range (Wu, Ma, Liao, & Lu, 2005); Thirdly, in the present time, we don’t At PRB-SCNU, each enclosure is 15 m2 (3.0 m wide, 5.0 m long, and have effective conservative measures to stop poaching in the wild. 1.5 m high). Experience indicates that an enclosure with these Hence, we think the best solution is to maintain the trade-confiscated dimensions can meet the 's demand for space. Pangolins can pangolins in captivity and establish an artificial population by captive survive, grow, and reproduce well in such an enclosure. This is similar breeding, and at the appropriate time, to release these rescued individuals or their offspring into wild areas where is proper by prudent evaluating. Given that, we established the Pangolin Research Base for Artificial Rescue and Conservation Breeding of South China Normal University (PRB-SCNU) in June 2010. The longest survival time for a captive Sunda pangolin is now more than 5 years, and 26 females have become pregnant and given birth at the PRB-SCNU, indicating great progress in rearing, breeding, and maintaining this species. In this paper, we discuss the techniques and management experience that we have obtained on housing, husbandry, and captive breeding of the Sunda pangolin at PRB-SCNU. This provides a reference for Sunda pangolin rescue and captive breeding, and a guide to the improvement of captive care and management of other pangolin species.

1.1 | Housing

The Sunda pangolin is a tropical species that needs a warm and humid climate and cannot tolerate extremely cold weather (Challender et al., FIGURE 1 Flaky and chapped skin of the captive Sunda pangolin 2012; Wilson, 1994). Thus, to control the artificial ambient climate, all MJ60 ZHANG ET AL. | 3 to Heath and Vanderlip's (1988) argument that Chinese pangolins (Manis pentadactyla) require a minimum of 10–12 m2. A nest box (0.8 m long, 0.8 m wide, and 0.6 m high) with an entrance 0.25 m long by 0.25 m wide is provided in one corner of each enclosure for captive pangolins to sleep in. Sunda pangolins often defecate in water (Challender et al., 2012) and a water pool is also provided (0.6 m long × 0.5 m wide × 0.3 m deep) in the enclosure. This water pool satisfies the animal's defecation and swimming needs. Pangolins dig with their strong foreclaws. However, their affect their walking posture, and they walk on their wrists by curling their long front digging claws inward and backward. Based on this, the nest boxes and enclosures have a concrete floor to prevent the captive pangolins from digging burrows and escaping, covered with a 0.1 m thick layer of sandy soil (sand: clay = 1:1) to prevent the pangolins from directly walking on the cement floor, which could lead to foreclaw injuries. Initially, wood shavings or straw were placed in the nest boxes to FIGURE 2 Soft feces of Sunda pangolin FS15 fed with fishmeal improve comfort and maintain the temperature, but these materials were easily eaten by the pangolins (Ogilvie & Bridgwater, 1967) and even led to three deaths from suffocation or gastric retention (Griner, not digest these . Based on the feces of pangolins fed with 1983; Zhang, Kong, et al., 2015; unpublished records at PRB-SCNU). grains, the food was barely digested (Figure 3) and the feces smelled a Subsequently, cotton rags with a diameter larger than 10 cm replaced little sour. We believe that the high mortality rate of captive pangolins the wood shavings and straw as a new bedding material, which and their susceptibility to gastrointestinal disease may be related to the resolved the fatalities resulting from the bedding material. inability of their digestive physiology to adapt to artificial diets. In an artificial environment, pangolins can show stereotypic Chitin is the second most abundant carbohydrate in nature behavior (Challender et al., 2012; Mohapatra & Panda, 2014). This may (Bussink, Speijer, Aerts, & Boot, 2007), making up 58–85% of the be related to the monotonous captive environment. Considering that of arthropods (Stevens & Hume, 1995). A study on high housing environmental heterogeneity is conducive to the digestive enzymes found that some insectivorous primates have a low expression of animal behavior, improving their ability to withstand multiple of forms of pepsinogen (Kageyama, 2000; Narita et al., 2000), the stress of surrounding factors (Carlstead, 1996), we use a wooden and chitinase enzymes are expressed in some myrmecophagous frame, some plastic pipes, and several large stones to enrich the captive species (Jeuniaux, 1962). Janiak (2016) suggested that insectivorous environment and provide the pangolins with climbing and shelter species may not require multiple pepsinogens to digest , but facilities. These facilities in the enclosure increase the welfare of the rather, express chitinolytic enzymes to digest insect exoskeletons. Diet captive pangolins. Activity and behavioral complexity improve, with no appears to be a driving factor of digestive enzymes and may have obvious stereotypic behavior. convergently evolved in sharing the same feeding habits (Janiak, 2016). Because pangolins feed on ants and termites, which have abundant chitin, we speculate that they and other myrmecoph- 1.2 | Artificial food and diet transition agous species have similar digestive enzymes, which may be why Pangolins have a highly specialized diet, consisting mainly of ants and pangolins cannot digest the meat of . termites (Wu, Liu, Li, & Sun, 2005; Lim, 2007), and Yang, Chou, and -based foods, particularly from insects, which are Chao (1999) suggest some crickets, flies, worms, earthworms, and relatively close to the natural diet, such as mealworm powder, other are also natural diet of pangolins. Under captive silkworm pupae powder, and fly maggot powder, are thought to be conditions, it is not feasible in the long term to provide sufficient more appropriate ingredients of artificial food than other materials. amounts of ants and termites to meet the requirements of captive These ingredients can be mixed together in appropriate proportions pangolins. Thus, supplying a suitable artificial food to meet the with some nutritional additives, such as multivitamin and mineral pangolins’ nutritional needs is very important. According to reports, supplements. At PRB-SCNU, the formulation of the artificial food was some institutions feed pangolins meat, eggs, dairy products, feline based on the above guidelines and practical experience. The effects of diets, and even apples and grains, such as corn, rice, and bread (Yang the artificial food were good. The pangolins defecated regularly and et al., 2007; Vijayan et al., 2008). The life span of pangolins is greatly the feces were solidly formed (Figure 4) with little inherent odor. A shortened in captivity, most dead pangolins had gastrointestinal digestion trial was performed for 8 days with six Sunda pangolins, and disease (Chin et al., 2012; Hoyt, 1987; Yang et al., 2001, 2007). This the results showed that the apparent digestibility coefficients of crude study found that pangolins had dyspepsia symptoms after eating the protein, crude fat, and dry matter were 81.92 ± 1.15%, 97.72 ± 0.85%, artificial foods mentioned above. For example, they had diarrhea after and 70.35 ± 4.12% (n = 6), respectively, indicating initial success in eating fishmeal (Figure 2) or eggs, and it was obvious that they could developing artificial pangolin food. 4 | ZHANG ET AL.

However, some pangolins refuse to eat fresh ants within 7 days after arrivalat PRB-SCNU. In this case, we force-feed them artificial food to keep them alive. The detailed method is as follows. In the evening, when the pangolin becomes active in the enclosure, we weigh artificial food at 10 g/kg body weight (about 2/3 of the normal feeding amount), mix it with a suitable amount of water to form gruel in a soybean milk machine, allow the pangolin to smell the fluid for a few minutes, and then cautiously inject the liquid food into the pharyngeal cavity using a medical injector. Compulsory feeding is stopped after 5–7 days and a thin paste is placed in a bowl to induce them to eat, while withholding water. Pangolins are thereby forced to eat the thin paste in the bowl to obtain water. Most pangolins eat the food within 3 days, although some individuals still refuse to eat. If so, we force feed them for another 3 days, and then entice them to eat until they take the food on their own. FIGURE 3 Feces of Sunda pangolin MJ5 fed with cereal grains A total of 41.1% of the incoming pangolins have finished the feeding transition by eating ants, 36.7% have needed fluid artificial Newly arrived pangolins are often stressed by capture and food, and 22.2% have died during the feeding transition (n = 158). At transportation and by sudden food changes, and may need to be PRB-SCNU, pangolins are considered to have adapted to the artificial encouraged to transition to the artificial food. Different institutions environment when stressed behavior disappears and the individual use different methods to induce them to eat. In the Oklahoma Zoo, appears healthy and freely eats artificial food (Zhang, Wu, et al., 2015). sawdust is used to lure Indian pangolins (M. crassicaudata) to eat (Ogilvie & Bridgwater, 1967), while torpid live ants are sprinkled on the 1.3 | Husbandry surface of artificial food to entice Sunda pangolins to eat at the Singapore Night Zoo (Vijayan et al., 2008). Pangolins are extremely difficult to maintain in captivity (Mohapatra & At PRB-SCNU, only freshly frozen Polyrhachis dives and clean water Panda, 2014; Yang et al., 2007), so it is very important to develop are provided to newly arrived pangolins. In this way, the pangolins can appropriate husbandry. The quality of management is directly related adapt to their new artificial environment as quickly as possible, the stress to the success or failure of the maintenance program. At PRB-SCNU, caused by the new environment is reduced, and a foundation is laid for husbandry includes cleaning enclosures, treating diseases in the further artificial food consumption. After arrival, some pangolins begin morning, and monitoring and adjusting the ambient temperature and to eat the ants within 5 days. Once a pangolin begins to eat fresh ants, humidity. We offer food between 18:00 and 23:00 every evening, and 20 g fresh ants per day are offered for the next 2 days and artificial food evaluate their health status daily via a monitor and direct observation. is gradually introduced on the third day. The amount is increased Captive pangolins are routinely physically examined and weighed once according to body size, but care is taken to avoid feeding too much. In a week. Regular deworming is performed quarterly and environmental this way, a good appetite can be maintained. All of the above benefit disinfection is conducted twice a week. feeding habit adjustment. Pangolins adjust to the artificial diet (i.e., feeding habit transition is completed) over 15–30 days. 1.4 | Feeding pattern

It has been reported that pangolins can be housed together in pairs or even as a small group (Challender et al., 2012; Nguyen Van, Clark, & Tran, 2010; Wang et al., 2015; Wilson, 1994; Yang et al., 1999). However, they are solitary, and males in close proximity may experience increased stress or increased mortality (Challender et al., 2012). Therefore, at PRB-SCNU all pangolins are housed individually in separate enclosures, except mating males and females and mother pangolins and their infants. This is similar to the procedure at Pangolin Conservation Breeding Centre (PCBC) of Nandankanan Zoological Park (NKZP) in India (Mohapatra & Panda, 2014).

1.5 | Control of temperature and humidity

As previously mentioned, the Sunda pangolin is sensitive to fluctua- FIGURE 4 Firm feces of Sunda pangolin MJ12 fed the artificial tions in ambient temperature and humidity. It is important to monitor PRB-SCNU food the ambient temperature and humidity and adjust them in a timely ZHANG ET AL. | 5 fashion. We monitor the ambient temperature and humidity daily. of their active time in behavior. This not only helps to increase Temperature is maintained below 33°C in summer. In autumn, winter, activity but also helps to maintain a diversity of behaviors in captivity. and spring, we pay close attention to changes in climate to prevent Every morning, we examine food intake and defecation, clean the large fluctuations in ambient temperature and humidity, maintaining enclosures, wash the food and water bowls, and record the data. them at over 20°C and 60%, respectively. We are able to provide a relatively constant artificial environment that is warm and humid. 1.8 | Monitoring of body weight

We weigh the pangolins weekly to monitor changes. This is important in 1.6 | Disease prevention managing captive pangolins, as feeding intake can be adjusted according Disease prevention is extremely important for captive animals. To to weightchanges andmonitoringprovides a basis for health assessment. prevent potential outbreaks of infectious diseases at PRB-SCNU, we In this way, we can detect and treat ill pangolins in a timely fashion. disinfect the captive enclosures inside and out as well as the surrounding research base, and change the bedding materials every 1.9 | Parasites and treatments week. Given that pangolins have an excellent , they may be sensitive to some odors. Hence, we do not use any chemicals with a When pangolins arrive at PRB-SCNU, we give them a medicated bath pungent smell (e.g., chlorine dioxide) and instead opt for an odorless of a povidone iodine solution (povidone iodine: water = 1:1500) and disinfectant. A 2% sodium hydroxide solution is used to sterilize the conduct a general physical examination after cleaning off surface dirt. outside of the pangolin enclosures and the surrounding areas around Most incoming pangolins have ectoparasites and evidence of trauma. the PRB-SCNU. The interior of each enclosure is sterilized with 0.1% However, they are extremely sensitive to some compounds, such as potassium permanganate. A 0.1% povidone iodine solution is used as a DDT and Lindane, and death can occur if these substances are used to disinfectant of feeding utensils, such as food and water bowls. remove ectoparasites (Van Ee, 1966). Instead, we remove ectopar- asites manually using tweezers and then treat the trauma. We find that newly captive Sunda pangolins may have intestinal 1.7 | Food intake control (Figure 5a) and mesenteric parasites (Figure 5b), which can lead to Generally speaking, both malnutrition and overnutrition are bad for death (Zhang, Kong, et al., 2015). Individuals are fed the broad- animal health. Initially, we fed Sunda pangolins, weighing between 2 kg spectrum repellent abamectin (Kunming Veterinary Medicine Co., and 4 kg, 20 g/kg body weight per day, but we found that individuals LTD., China) at a dose of 400 mg/kg body weight, which is effective for fed this amount were obviously fatter than those who had just arrived elimination of internal parasites. Routine deworming is performed at PRB-SCNU. This may be related to the low metabolism of pangolins quarterly using abamectin at the initial dose for 3 days. (Heath, 1987; Lin, Chang, Yang, & Dierenfeld, 2015). To avoid obesity, Sunda pangolins, Chinese pangolins, and Indian pangolins all suffer we have to control the food intake of the captive animals. During from ectoparasites and endoparasites. Amblyomma javanense is an husbandry, we found that an increase in body weight of 10 g a day was ectoparasite of these three pangolin species (Hassan, Sulaiman, & Lian, appropriate for pangolin health. Thus, according to their growth stage 2013; Sanyal, De, Rao, & Acharjyo, 1987; Yang, Su, Zhang, Wu, & Ma, and daily food intake, we feed the pangolins artificial food at 15–20 g/ 2010). Chinese pangolins and Indian pangolins can be infected by kg body weight per day to individuals weighing less than 2.5 kg and nematodes and other types of parasites (Heath & Vanderlip, 1988; feed 12–18 g/kg body weight per day to those weighing 2.5–5.0 kg. Mohapatra & Panda, 2014). However, we have not identified the Sunda pangolins are nocturnal animals, usually foraging, drinking, ectoparasitic and endoparasitic species collected from Sunda pangolins. urinating, and defecating between 17:00 and 08:00 (Challender et al., 2012; Lim, 2007; Wang et al., 2015). To reduce the frequency of 1.10 | Physical examination stereotypic behavior, fresh food and clean water are provided at a random time between 18:00 and 23:00, and placed at a random During captivity, we qualitatively observe behavior, eating, and location in the enclosure. This encourages the pangolins to spend some defecation every day, check for parasites in feces, and perform a

FIGURE 5 Intestinal (a) and mesenteric parasites (b) of Sunda pangolins 6 | ZHANG ET AL.

FIGURE 6 A male Sunda pangolin mating with a female in captivity

TABLE 1 Records of captive breeding of Sunda pangolins at PRB-SCNU Mother Date arrived in Father Parturition Gestation period Baby No. ID captivity ID Mating date date (day) Baby ID gender 1 QF31 Nov.?, 2012 QF9 ? Jan. 10, 2014 ? QFA4 ♂ 2 QF18 Nov.?, 2012 QF29 ? Jan. 14, 2014 ? QFA5 ♂ 3 MJ-X1γ Bef. May 1, 2012 ? Jul 16–Sep 8, 2012 Jan. 23, 2013 137–191 MJ-X1b ♂ 4 MJ65γ Jun. 10, 2011 MJ66 Aug 15–Oct 2, Feb. 14, 2012 135–183 MJ132* ♀ 2011 5 MJ60γ May 12, 2011 MJ71 Aug 19–Nov 10, Mar. 10, 2012 120–203 MJ133* ♀ 2011 6 QF41 Dec.?, 2012 QF12 ? Mar. 21, 2014 ? QFA6 ♂ 7 MJ-X2γ Bef. May 1, 2012 MJ120 Sep 10–Nov 15, Mar. 27, 2013 134–200 MJ-X2b ♀ 2012 8 MJ-X3γ Bef. May 1, 2012 MJ120 Sep 10–Nov 15, Apr. 12, 2013 150–216 MJ-X3b ♀ 2012 9 QF54 Dec.?, 2012 QF12 ? Apr. 22, 2014 ? QFA8 ♀ 10 QF57 Dec.?, 2012 QF9 ? May 4, 2014 ? QFA9 ♀ 11 QF40 Dec.?, 2012 QF29 ? May 5, 2014 ? QFA10 ♀ 12 QF2 Aug.?, 2012 QF37 ? Jun. 20, 2014 ? QFA11 ♀ 13 QF47 Dec.?, 2012 QF29 ? Jun. 21, 2014 ? QFA12 ♂ 14 QF33 Nov.?, 2012 QF12 ? Jul. 9, 2014 ? QFA13 ♀ 15 QF14 Sep.?, 2012 QF9 ? Jul. 9, 2014 ? QFA14 ♀ 16 QF6 Sep.?, 2012 QF60 ? Sep. 10, 2014 ? QFA15 ♀ 17 MJ75γ Jun. 10, 2011 MJ71 Mar 28–Apr 15, Sep. 20, 2012 158–176 MJ188* ♂ 2012 18 QF17 Nov. 9 ∼ 11, 2012 QF12 ? Sep. 29, 2014 ? QFA16 ♀ 19 MJ90γ Dec. 14, 2011 MJ66 Mar 28 & Apr 4, Oct. 2, 2012 181 or 188 MJ189*λ ♂ 2012 20 QF7 Aug.?, 2012 QF15 ? Oct. 30, 2014 ? QFA18*λ ♂ 21 QF69 Dec.?, 2012 ? ? Oct.?, 2014 ? QFA17* ? 22 QF50 Dec.?, 2012 QF29 ? Nov. 28, 2014 ? QFA19 ♀ 23 QF58 Dec.?, 2012 QF9 Jun 11–13, 2013 Dec. 12, 2013 182–184 QFA1 ♂ 24 QF50 Dec.?, 2012 QF29 Jun 24–27, 2013 Dec. 19, 2013 175–178 QFA2 ♂ 25 QF1 Aug.?, 2012 QF12 Jun 20–24, 2013 Dec. 20, 2013 179–183 QFA3 ♂ 26 QF43 Dec.?, 2012 QF37 Jun 20–24, 2013 Dec. 23, 2014 182–186 QFA20* ♀

*, stillbirth; λ, the babe with malformation; ?, data no available; γ, date from Zhang, Wu, et al. (2015). ZHANG ET AL. | 7 complete physical examination every week. These procedures are the pair is separated and the female is housed individually. Based on important in diagnosing and treating sick animals in a timely fashion. female weight gain and appetite, more food is provided along with Healthy Sunda pangolins have a good appetite, normal activity, and more vitamins and minerals to meet her nutritional needs during defecate regularly. There should be no injuries or sores on their skin pregnancy until parturition. and no secretions from the mouth, eyes, ears, arms, or genitals. Body The gestation time of Sunda pangolins is about 6 months (n =6, temperature is about 30–35°C, similar to healthy captive Indian Table 1). When the pangolins are about to give birth, feeding pangolins (Mohapatra & Panda, 2014). If a pangolin is ill, one or more of management is improved and activity is observed closely. However, the following may be observed: low activity, loss of appetite, vomiting, due to lacking of the monitor in some enclosures, and some females diarrhea, unusual secretions, and reduced body temperature. giving birth to cub in the nest, only four cases of delivery were Given that pangolins are inactive during the day, treatment of observed, although 34 Sunda pangolin cubs have been born at PRB- wounded pangolins in the morning is more effective in promoting SCNU. Before giving birth, pregnant pangolins show a series of healing than treatment at other times. To treat some common diseases, abnormal behaviors such as anxiety, walking rapidly around the some medications have been shown to be safe and effective, for enclosure with occasional stops and looking at the abdomen, digging at instance, 80.72% of pangolins have been cured by injecting kanamycin the base of the enclosure, and moving of feeding utensils. During at 0.2 ml/kg body weight; of 76 pangolins with pneumonia treated with delivery, females face the wall in a corner, their forelimbs slightly off injections of lincomycin hydrochloride at 0.3 ml/kg body weight, 65 the ground, and move into a defecating position, accompanied by individuals recovered. regular abdominal contractions, they finally give birth their babies within 209.00 ± 56.81 min (145–270 min, n = 4; Figure 7). After the cub has been born, both female and cub are housed in the same 1.11 | Captive breeding enclosure. Human disturbance is avoided. The mother and cub are Although many zoos and other institutions keep pangolins in captivity, housed separately after weaning at about 4 months (Lim & Ng, 2008; only the Singapore Zoo and the PRB-SCNU have bred them in captivity Zhang, Wu, et al., 2015). (Zhang, Wu, Yang, Li, & Zhang, 2013; Zhang, Yang. Wu, Li, & Zhang, In this study, to date, a total of 43 Sunda pangolin births were 2013; Zhang, Wu, et al., 2015). In other cases, the females were already recorded (Challender et al., 2012; Lim & Ng, 2008; Wang, 2015; Yu pregnant before they arrived in the artificial environment (Challender et al., 2016; Zhang, Wu, et al., 2015; unpublished records at PRB- et al., 2012; Wang, 2015; Yang et al., 2007). SCNU), 34 Sunda pangolin cubs have been born at PRB-SCNU, 26 cubs At PRB-SCNU, after Sunda pangolins adapt to the captive who born at PRB-SCNU were bred in captivity, of which 19 were environment, one female and one male are selected at irregular healthy and 7 died (Table 1). Births have been recorded throughout the intervals and housed together for mating, and their sexual responses year, with the exception of August (n = 43; Figure 8). In this study, it is are observed. The selected pairs must be deemed healthy, weigh above the first report that Sunda pangolins can birth in May and December., 3.0 kg to ensure that they have reached sexual maturity (Yang et al., Zhang, Wu, et al., (2015) inferred that sexual maturity of the Sunda 2010), and the male should be bigger than the female to grasp the latter pangolin occurred at approximately 1 year, even it could occur as early for mating. Mating occurs many times during the joint housing period as 6–7 months. Always only one offspring was born at a time (n = 43). of several days. Thus, if the male and female pangolins are observed to mate successfully (Figure 6), they will be housed together for 5–7 days to increase the probability of successful mating and pregnancy. Then,

FIGURE 8 Numbers of births of Sunda pangolins in different months throughout the year (2002–2015). Data from Lim and Ng (2008), Challender et al. (2012), Yu et al. (2016), Wang (2015), Zhang, Wu, et al., 2015, Singarpore Zoo, Wildlife Rescue Center of FIGURE 7 Newborn baby MJ88 and its mother MJ70 Guangdong Province, and PRB-SCNU 8 | ZHANG ET AL.

Weaning occurred at about 3–4 months (Challender et al., 2012; Lim & 2 | CONCLUSIONS Ng, 2008; Zhang, Wu, et al., 2015). Pangolins are the most illegally trafficked wild animals in the world. It is estimated that more than 1 million pangolins have been killed in 1.12 | Causes of death the last decade, resulting in the depletion of wild pangolins, and the In total the deaths of 93 Sunda pangolins have been dissected and safety of the remaining individuals in the wild is uncertain. At PRB- recorded at PRB-SCNU. Pneumonia and gastrointestinal issues were SCNU, we rescue pangolins confiscated from illegal trade to develop the main diseases seen (Figure 9). In addition, complete loss of techniques to maintain and breed captive pangolins, establish a appetite, hepatitis, nephritis, skin disease, and endoparasites were viable artificial population, and release some individuals to rehabil- major health concerns (Figure 9). This is similar to other pangolin itate wild pangolin populations after the artificial population reaches species when kept in captivity (Hoyt, 1987; Hua et al., 2015; a certain size. Our experience indicates that providing Sunda Mohapatra & Panda, 2014; Yang et al., 2007). Choo et al. (2016) pangolins an enclosure of 12–15 m2, enriching the captive environ- found that the IFNE gene, a unique interferon expressed only in skin ment, maintaining the temperature between 20 and 33°C and epithelial cells and inner mucosa-protected tissues, is pseudogenized humidity between 60% and 90%, and providing an appropriate in Sunda pangolins, suggesting that innate immunity may be artificial food and appropriate veterinary care allow them to survive, compromised. Furthermore, captive pangolins are prone to infections grow, develop, and breed well in captivity. Additional studies are by pneumonia and gastrointestinal diseases. Based on captive needed on the nutrient and environmental needs of the Sunda individuals, it is thought that disease incidence is related to pangolin to improve the artificial food and environment. This will environmental stress, ambient temperature and humidity, and artificial greatly improve the welfare of captive pangolins, reducing morbidity food. For example, some captive pangolins display pica, ingesting the and extending their lifespans. sandy soil in their enclosures (e.g., MJ12, DY8, and DY9). This phenomenon is also seen in some other myrmecophagous mammals ACKNOWLEDGMENTS (Gull et al., 2015; Lin et al., 2015). Ingestion of soil may be related to a lack of certain nutrients or pH adjustment of the gut (Gull et al., 2015). We would like to thank all those who work at the Pangolin Research However, little research has been conducted on this. Therefore, Base for Artificial Rescue and Conservation Breeding of South China continuing to survey the feeding habits of pangolins and further Normal University (PRB-SCNU) for their help in data collection. We research on the nutrient needs of pangolins will help to develop an would further like to acknowledge and thank the Wildlife Rescue artificial food that can better meet the needs of pangolins in captivity. Center of Guangdong Province and forestry public security of Maoming city (Guangdong Province) for providing the animals for this study. This research was supported by the Natural Science Foundation of Guangdong Province (No. S2013010013356), the Special Fund for Significant Science and Technology in Social Development Field of Guangdong Province (No. 2011A030100012), the National Natural Science Foundation of China (31572286, 31670266, 31702029), Project supported by Guangdong Province Universities and Colleges Pearl River Scholar Funded Scheme (2012), and the public program ‘Conservation Action for Critically Endan- gered Pangolins’ funded by China Environmental Protection Foundation.

ORCID

Fuhua Zhang http://orcid.org/0000-0002-5179-386X

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