NOTE Public Health Prevalence of Salmonella spp. in Pet Reptiles in Japan Aya NAKADAI1,2), Toshiro KUROKI3), Yukio KATO2), Rieko SUZUKI3), Shiro YAMAI3), Chiharu YAGINUMA2), Ryo SHIOTANI4), Akira YAMANOUCHI5) and Hideki HAYASHIDANI1)* 1)Division of Animal Life Science, Institute of Symbiotic Science and Technology, Tokyo University of Agriculture and Technology, 3–5–8 Saiwai-cho, Fuchu, Tokyo 183–8509, 2)Department of Public Health II, School of Veterinary Medicine, Azabu University, 1–17–71 Fuchinobe, Sagamihara, Kanagawa 229–8501, 3)Kanagawa Prefectural Institute of Public Health, 1–3–1 Shimomachiya, Chigasaki, Kanagawa 253–0087, 4)Gentleman Loser, 5–29–7 Fuchinobe-honcho, Sagamihara, Kanagawa 229–0002 and 5)Yamanouchi Iguana Laboratory, 103 Lions Plaza 7–5–8 Sagami-ono, Sagamihara, Kanagawa 228–0803, Japan (Received 28 May 2004/Accepted 27 August 2004) ABSTRACT. From November 2000 to July 2002, 112 fecal samples from pet reptiles, including 18 turtles, 71 lizards and 23 snakes, sold at a pet shop were examined for the prevalence of Salmonella spp. in Japan. Salmonella spp. were isolated from 83 (74.1%) of 112 samples, and a total of 112 Salmonella isolates were identified as subspecies I to IV. The majority of isolates (62.5%) belonged to sub- species I and 54 isolates could be identified as any of 28 serovars. The predominant serovars were found to be S. Bardo, S. Newport and S. Panama, which cause human salmonellosis. These results indicate that pet reptiles may be a potential infectious source of human salmonellosis in Japan. KEY WORDS: prevalence, reptiles, Salmonella. J. Vet. Med. Sci. 67(1): 97–101, 2005 Salmonellosis is known to be one of the most important Carry-Blair transport medium (Eiken Chemical Co., Ltd., cause of public health problems worldwide. In Japan, more Tokyo, Japan), and immediately transported to the labora- than 460 cases of human salmonellosis occurred in 2002, tory in an icebox. The fecal samples were suspended in 3 ml and most of them resulted from consumption of foods of of sterile saline, and 1 ml of the suspension was inoculated animal-origin contaminated with Salmonella [14]. Wild and into 10 ml of buffered peptone water (BPW, OXOID Ltd., pet reptiles are generally known to be asymptomatic carriers Basingstoke, Hampshire, England). After incubation at of several Salmonella serotypes, which are potentially 37°C for 24 hr, 1 ml of BPW culture was transferred to 10 pathogenic for humans. Recently, the number of exotic rep- ml of Hajna-tetrathionate broth (Eiken). The broth was tiles has been increasing in popularity as pets and the fact incubated at 37°C for 24 hr, then one loopful of each tube has led to an increase in the number of reptile-associated was inoculated onto a plate of brilliant green agar (BGN, Salmonella infections in the United States [4–6, 18] and OXOID) supplemented with 20 µg novobiocin/ml, desoxy- European countries [2, 7–9]. Majority of the patients cholate hydrogen sulfide lactose agar (DHL, Nissui Pharma- infected with Salmonella spp. from reptiles are infants, and ceutical Co., Ltd., Tokyo) and mannitol lysine crystal violet usually show gastroenteritis, less frequently, fatal meningi- brilliant green agar (MLCB, Nissui). These plates were tis or septicemia [5,7,8]. Recently, many reptiles have also incubated at 37°C for 24 hr and three suspicious colonies been kept as pets in Japan as well as in other developed morphologically similar to Salmonella spp. from each plate countries and it is possible that these animals can be a source were subcultured for biochemical examinations. Biochemi- of human salmonellosis. However, little is known about the cal characteristics were examined on triple sugar iron prevalence of Salmonella spp. in reptiles except for aquar- medium (Nissui), lysine indole motility medium (Nissui) ium turtle in Japan [13,19]. This study was designed to and Voges-Proskauer semisolid medium (Eiken). The sub- investigate the presence of Salmonella in pet reptiles, sold species of Salmonella isolates was confirmed by biochemi- and kept in Japan. cal examinations according to Holt et al. [12]. Serotyping of During the period from November 2000 through July Salmonella isolates was accomplished with commercial O 2002, a total of 112 fecal samples of 50 reptile species con- and H antisera (Denka Seiken Co., Ltd., Tokyo) according sisting of 12 families were collected (Table 1). All reptiles to the method of Popoff and Le Minor [16]. except green iguanas were sold as pets in a pet shop located Salmonella spp. were isolated from 83 (74.1%) of 112 in Kanagawa Prefecture, and green iguanas were kept at fecal samples of reptiles (Table 1). Salmonella spp. were Yamanouchi Iguana Laboratory. Reptiles sold in the shop isolated from 23 (100%) of 23 snakes, 13 (72.2%) of 18 tur- had been kept in individual cages immediately after they tles, and 47 (66.1%) of 71 lizards. The isolation rate from were introduced into the shop. Fecal samples were put into snakes was the highest of all. This high proportion of Sal- monella infection in captive reptiles, especially snakes, is *CORRESPONDENCE TO: HAYASHIDANI, H., Division of Animal Sci- ence, Institute of Symbiotic Science and Technology, Tokyo similar to the results of several previous studies [10, 15, 18]. University of Agriculture and Technology, 3–5–8 Saiwai-cho, Geue et al. [10] examined the prevalence of Salmonella in Fuchu, Tokyo 183–8509, Japan. pet reptiles in Germany and Austria, and reported that Sal- 98 A. NAKADAI ET AL. Table 1. Distribution of samples examined, number of isolates and serovars of Salmonella associated with reptile species Reptile taxonomy No. of samples (%)a) No. of Subspecies and Serovars Common name Nomenclature isolates (No. of isolates) Order Testdines (Turtles) 13/ 18 (72.2) 20 Family Testdidae Red-footed Tortoise Geochelone carbonaria 0/ 1 (0.0) 0 – Indian Starred Tortoise Geochelone elegans 1/ 1 (100.0) 1 Salmonella subsp.I (UTb)) (1) Leopard Tortoise Geochelone pardaris 7/ 8 (87.5) 11 S. Minnesota (3) S. Montevideo (1) Salmonella subsp.I (UT) (5) Salmonella subsp.IIIb (UT) (2) Greace Tortoise Testudo graeca ibera 1/ 1 (100.0) 1 S. Bareilly (1) Central Asian Tortoise Testudo horsfieldi 1/ 1 (100.0) 3 S. Horsham (1) S. 13:z29:1,5 (subsp.II) (1) Salmonella subsp.II (UT) (1) Southern Spider Tortoise Pyxis arachnoides oblonga 0/ 1 (0.0) 0 – Madagascar Flat-shelled Tortoise Pyxis planicauda 0/ 1 (0.0) 0 – Family Emydidae Three-toad Box Turtle Terrapene carolina 2/ 2 (100.0) 2 Salmonella subsp.I (UT) (2) Family Chelydridae Snapping Turtle Chelydra serpentina 0/ 1 (0.0) – Alligator Snapping Turtle Macroclemys temminckii 1/ 1 (100.0) 2 S. Haifa (1) S. Potsdam (1) Order Squamata Suborder Sauria (Lizards) 47/ 71 (66.1) 57 Family Agamidae Inland Bearded Dragon Pagona vitticeps 15/ 16 (93.8) 16 S. Amsterdam (1) S. Beaudesert (1) S. Kisarawe (1) S. Minnesota (1) S. Rissen (1) S. 16:z4: – (subsp.IV) (1) Salmonella subsp.I (UT) (3) Salmonella subsp.II (UT) (5) Salmonella subsp.IV(UT) (2) Egyptian Spiny-tailed Agama Uromastyx aegypticus 1/ 1 (100.0) 1 Salmonella subsp.II (UT) (1) Indian Spiny-tailed Agama Uromastyx hardwickii 3/ 4 (75.0) 3 Salmonella subsp.I (UT) (3) Family Iguanidae Green Iguana Iguana iguana 3/ 12 (25.0) 3 Salmonella subsp.IV (UT) (3) Family Scincidae Monky-tailed Skink Corucia zebrata 0/ 1 (0.0) 0 – Pygmy Spiny-tailed Skink Egernia depressa 1/ 2 (50.0) 1 Salmonella subsp.I (UT) (1) Family Cordylidae Rough-scaled Plated Lizard Gerrhosaurus major 2/ 2 (100.0) 2 S. Amsterdam (1) Salmonella subsp.I (UT) (1) Family Helodermatidae Banded Gila Monsters Heloderma S. cinctum 1/ 1 (100.0) 1 S. Poona or Farmsenc) (1) Reticulated Gila Monsters Heloderma S. Suspectum 1/ 1 (100.0) 2 S. Alachua (1) Salmonella subsp.I (UT) (1) Family Varanidae Gray’s Monitor Varanus olivaceus 1/ 1 (100.0) 3 S. Haifa (1) Salmonella subsp.I (UT) (1) Salmonella subsp.IIIb (UT) (1) Roughneck Monitor Varanus rudicllis 1/ 1 (100.0) 2 S. Kentucky (1) S. Schwarzengrund (1) Family Gekkonidae Texas Banded Gecko Coleonyx brevis 1/ 2 (50.0) 1 S. Beaudesert (1) Central American Banded Gecko Coleonyx mitratus 0/ 2 (0.0) – Malayan Bow-fingered Gecko Cyrtodactylus pulchellus 0/ 1 (0.0) – Leopard Gecko Eublepharis macularius 1/ 5 (20.0) 1 S. Amsterdam (1) Japanese Gecko Gekko japonicus 1/ 1 (100.0) 2 S. Bardo (1) S. Panama (1) Continued on following page. SALMONELLA FROM PET REPTILES 99 Table 1. – Continued. Reptile taxonomy No. of samples (%)a) No. of Subspecies and Serovars Common name Nomenclature isolates (No. of isolates) Indian House Gecko Hemidactylus flaviridas 1/ 1 (100.0) 1 Salmonella subsp.IIIb (UT) (1) Masobe Madagascar Ground Gecko Paroedura masobe 1/ 1 (100.0) 1 S. Enteritidis (1) Madagascar Ground Gecko Paroedura pictus 6/ 9 (66.7) 8 S. Hvittingfos (1) S. Panama (1) S. Potengi (1) S. Soananina or Sundsvallc) (2) Salmonella subsp.I (UT) (1) Salmonella subsp.II (UT) (2) Carter’s Rock Gecko Pristurus rupestoris 1/ 1 (100.0) 1 S. Stanley (1) Crested Gecko Rhacodactylus ciliatus 3/ 3 (100.0) 4 S. Braenderup (2) Salmonella subsp.IV (UT) (2) Carrot-tailed Viper Gecko Teratolepis fasciata 2/ 2 (100.0) 2 S. Panama (2) Fantastic Flat-tailed Gecko Uroplatus phantasticus 1/ 1 (100.0) 2 S. Muenchen (1) Salmonella subsp.IIIb (UT) (1) Order Squamata Suborder Ophidia (Snakes) 23/ 23 (100.0) 35 Family Boidae Children’s Python Antaresia childreni 1/ 1 (100.0) 1 Salmonella subsp.II (UT) (1) Garden Tree Boa Corallus hortulaus 2/ 2 (100.0) 3 S. Kentucky (1) Salmonella subsp.IIIb (UT) (2) Kenyan Sand Boa Gongylophis colubrinus 1/ 1 (100.0) 1 Salmonella subsp.IIIb (UT) (1) Green Python Morelia viridis 1/ 1 (100.0) 1 S.