Gastrointestinal Helminths of Feral Raccoons (Procyon Lotor) in Wakayama Prefecture, Japan

FULL PAPER Parasitology

Gastrointestinal Helminths of Feral Raccoons (Procyon lotor) in Wakayama Prefecture, Japan

Hiroshi SATO1) and Kazuo SUZUKI2)

1)Laboratory of Veterinary Parasitology, Faculty of Agriculture, Yamaguchi University, 1677–1 Yoshida, Yamaguchi 753–8515 and 2)Hikiiwa Park Center, 1629 Inari-cho, Tanabe 646–0051, Japan

(Received 30 May 2005/Accepted 1 December 2005)

ABSTRACT. The population and distribution of feral raccoons (Procyon lotor) are expanding in Japan after escape or release from animal- owners. Wakayama Prefecture is one of the most typically devastated areas by this exotic carnivore, particularly in the last five years after a latent distribution for more than ten years. Official control measures of feral raccoons commenced in the summer of 2002 by several municipalities, and 531 animals collected in 12 municipalities between May 2003 and April 2005 were submitted for parasitological examination of gastrointestinal helminths. Detected parasites included six nematodes (Physaloptera sp. [prevalence; 5.1%], Con- tracaecum spiculigerum [0.9%], Strongyloides procyonis [25.5%], Ancylostoma kusimaense [0.8%], Arthrostoma miyazakiense [0.4%], and Molineus legerae [1.1%]), seven trematodes (Isthmiophora hortensis [4.9%], echinostomatid sp. with 34–39 collar spines [1.7%], Metagonimus takahashii [12.4%], M. yokogawai [0.8%], Plagiorchis muris [0.2%], Macroorchis spinulosus [1.9%], and Consinium ten [0.2%]), one cestode (Mesocestoides sp. [0.2%]), and six acanthocephalan spp. (Centrorhynchus bazalenticus [0.2%], Centrorhynchus teres [5.5%], Sphaerirostris lanceoides [2.4%], Plagiorhynchus ogatai [0.6%], Porrorchis oti [1.5%], and Southwelina hispida [1.9%]). Most of the collected parasites are food-borne, indigenous helminth species. Physaloptera sp. has never been recorded in indigenous wild carnivores in Japan, and resembles closely P. rara, prevalent in raccoons of North America, in morphology. The position of a pair of phasmids in the posteroventral region of the adult male, however, could differentiate it from P. rara. Since Strongyloides procyonis is known to cause creeping eruption as well as intestinal infection in a healthy human volunteer, we should be concerned about the rapid increase in the population and distribution of feral raccoons in Japan from the viewpoint of public health as well. KEY WORDS: gastrointestinal helminth, Physaloptera, Procyon lotor, Strongyloides procyonis, zoonosis. J. Vet. Med. Sci. 68(4): 311–318, 2006

The population and distribution of feral raccoons (Pro- cultural loss and housebreaking, an official control and cyon lotor (Linnaeus, 1758)) in Japan are expanding after eradication program of feral raccoons commenced in the intentional or unintentional releases by animal-owners. summer of 2002 in several municipalities of Wakayama Wakayama Prefecture is one of the most typically devas- Prefecture, including Tanabe City, which has been since tated areas in this country. Miyashita [27] recorded the dis- then adopted by other municipalities. tribution of feral raccoons in 15 prefectures in Japan, Raccoons adapt to a variety of habitats, and are often including Aichi, Gifu, Hokkaido, Kanagawa and attracted to areas of human activity by food available in the Wakayama, in March 1992. Earlier data on feral raccoons farmland and garbage in the town, or by spaces under the in Wakayama Prefecture is not available. Based on a recent roof that may offer shelter [16, 24]. Considering that serious inquiry conducted by the prefectural office (August 2003; zoonoses such as leptospirosis, tularemia, rabies, cryptospo- unpublished), a chronological change in the distribution of ridiasis and Baylisascaris procyonis larva migrans, and dis- raccoons in this prefecture is as follows (50 municipalities eases to companion animals such as canine and feline in total before 1 October 2004 or the great municipal merger distemper could be transmitted by raccoons [16, 24, 47], in Heisei era [Heisei shi-cho-son-gappei], and we follow effective control measures of feral raccoons are necessary this division thoroughly in descriptions of the present not only due to direct nuisance behaviors to human activities study): feral raccoons were noticed in 5 municipalities and disturbance of local ecosystem, but also to prophylacti- (Tanabe, Nakahechi, Kamitonda, Shirahama, and Kawabe) cally remove possible health threats to residents and indige- by 1997; another 5 municipalities located in the northern nous wildlife and domestic animals. In fact, recent region of this prefecture were added to the distribution area outbreaks of fatal neurological larva migrans caused by the by 1999; feral raccoons were noticed in 21 municipalities in raccoon roundworm B. procyonis in rabbits and Japanese total by 2001; and at least in 36 municipalities by 2003. At macaques kept in zoo facilities in Japan were serious [10, present, feral raccoons are widely distributed in at least 12 38, 41, 44], emphasizing the need for pet and feral raccoons municipalities after the inquiry data mentioned above (Fig. to be under strict control of public and animal health author- 1), and the agricultural loss caused by these animals ities. increased abruptly in the last five years: the area of damage In the present study, we examined gastrointestinal helm- was approximately 10 and 142 ha, resulting in economic inths of 531 feral raccoons collected in 12 municipalities of loss amounting to 1.5 and 32.9 million JPY, in 2001 and Wakayama Prefecture to assess their possible health threats 2002, respectively. Due to apparent nuisances such as agri- to residents and indigenous animals, with special attention 312 H. SATO AND K. SUZUKI

Fig. 2. Relationship between body weight and month of trapping of feral raccoons examined in the present study. The survey period is May 2003 to April 2005.

was opened longitudinally, scratched carefully by glove- worn hands, and the content was washed repeatedly by sim- ple sedimentation in tap water. Sediments were checked thoroughly under a dissection microscope. Except for several worms for DNA extraction, the collected parasites were fixed in 10% neutral-buffered formalin or 70% alcohol. Parasitological examination was carried out on 245 (100 female and 145 male) and 286 (130 female and 156 male) Fig. 1. Geographic distribution of feral raccoons examined in the feral raccoons collected in 12 municipalities during the first- present study. Wakayama Prefecture, Japan, was divided into 50 municipalities (exactly saying, this division was correct year (May 2003—April 2004), and second-year period before 1 October 2004 or the great municipal merger in Heisei (May 2004—April 2005), respectively. Geographical dis- era, but we followed this division here), and 531 examined rac- tribution and monthly numbers of feral raccoons examined coons in total were trapped in Hashimoto (HSM), Yura (YRA), in the present study are shown in Figs. 1 and 2. Body weight Gibo (GBO), Kawabe (KWB), I’nami (INM), Minabe (MNB), (BW) partially reflects the age of the animal [16], and Tanabe (TNB), Nakahechi (NHC), Kamitonda (KTD), Ohtoh roughly saying, raccoons less than 2.0 kg BW were juvenile, (OHT), Shirahama (SRH), and Hikigawa (HKG). Numbers in those between 2.0 and 4.0 kg BW growing young, and those parentheses after each abbreviated municipality name indicate the number of examined raccoons. Currently, feral raccoons are more than 4.0 kg BW young and grown adults. noticed in 36 municipalities (marked by dots or dark), and are DNA extraction and polymerase chain reaction (PCR): widely distributed in at least 12 municipalities (dark). Molecular genetic characterization was conducted on echinostomatid trematodes to identify the species [20, 28–30]. to the raccoon roundworm and other exotic helminth spe- Amplified were segments of small and large subunit (SSU/ cies. We believe that this kind of assessment of possible LSU) ribosomal RNA genes (rDNA) with internal tran- biohazards is important also for non-specialized workers scribed spacer 1 (ITS-1) and 2 (ITS-2), and mitochondrial engaged in the control of this animal at present as well as in genes (mtDNA) such as cytochrome c oxidase subunit I the future. (COI) and nicotinamide adenine dinucleotide dehydroge- nase subunit 1 (ND1). Parasites of interest were collected as MATERIALS AND METHODS promptly as possible, and preserved in 1.5 ml Eppendorf tubes at –80°C. DNA extraction, PCR, and DNA sequenc- Parasitological examination: Raccoons were trapped ing were conducted as described in our previous work [42]. using metal cage-traps by licensed residents, euthanized Each cycle of PCR consisted of denaturation for 30 sec at using a humanitarian method, labeled individually, and 94°C, annealing for 60 sec at 62–65°C (rDNA), 52°C (COI stored individually at –20°C until zoological examination. mtDNA), or 48°C (ND1 mtDNA), and extension for 60 sec After thawing, data related to sex, age class and zoological at 72°C. This cycle was repeated 32 times. The used prim- measurements were recorded. The entire intestine distal to ers were universal ones described by others [8, 28–30, 52]. the pylorus was removed from the carcass, refrozen, and DNA sequences were aligned using the CLUSTAL W mul- sent to the laboratory for parasitological examination. The tiple alignment program [49] available on the net [clust- stomach was opened to examine food items, and the col- alw.genome.jp/]. Nucleotide sequences reported here are lected parasites were fixed in 70% alcohol. The intestine deposited and available in the DDBJ/EMBL/GenBank as HELMINTHS OF FERAL RACCOONS IN JAPAN 313 mentioned below. 4 species, and 1 with 5 species. Infection with soil-transmitted nematodes of indigenous carnivores, such as Ancylos- RESULTS toma kusimaense, Arthrostoma miyazakiense, and Molineus legerae was rare in feral raccoons, and observed mainly in General view: Except for Strongyloides procyonis, which juvenile or younger raccoons with BW less than 4 kg. In was detected for the first time in the second-year period of addition, the number of these parasites from an individual our survey and then often found in raccoons examined, host was substantially small (Table 1). Examined raccoons recovery of other parasites showed no apparent differences had no roundworms including Baylisascaris procyonis, or by year. Table 1 summarizes the results of the parasite Toxocara tanuki that was found rarely from this host species recovery by host sex and body weight. The proportion of in Japan [4]. infected raccoons with any helminth species was higher in Physaloptera sp.: The adults measuring 2–4.5 cm in males than females regardless of BW (age), and increased length (Fig. 3), were found firmly attached to the gastric steadily with BW (age). Infection with a small number of a mucosa or sometimes embedded in the mucosal nodes in 24 single species was common, and infection with multiple raccoons, and a few juveniles were found in the small intes- species was seen only in 59 raccoons (27.4% of all infec- tine of 3 raccoons caught between May and July. Infected tions): 37 raccoons with 2 species, 16 with 3 species, 5 with raccoons were trapped mainly in Tanabe (16 cases), particu-

Table 1. Gastrointestinal helminths collected from feral raccoons in Wakayama Prefecture, Japan, between May 2003 and April 2005

Body weight (F, female; M, male) Parasite <2.0kg ≥2–<4 kg ≥4–<6 kg ≥6 kg Total F M F M F M F M Prevalence Range Locality of Number of animals examined 40 35 50 40 97 112 43 114 531 (%) in intensity infected raccoons* Number of animals infected 7 10 15 15 37 55 16 60 215 with any parasites (%) (17.5)(28.6) (30.0)(37.5) (38.1) (49.1)(37.2)(52.6) (40.5) Nematoda Family Physalopteridae Physaloptera sp. 0 1 2 1 9 7 2 5 27 5.1 1 – 57 TNB, HKG, SRH, KTD, INM, KWB Family Anisakidae Contracaecum spiculigerum 000002125 0.93 TNB Family Strongyloididae Strongyloides procyonis 2 3 5 7 18 14 10 14 73 25.5** 1 – 197 TNB, HKG, SRH, KTD, INM, KWB, GBO,YRA Family Ancylostomatidae Ancylostoma kusimaense 030100004 0.81 TNB Arthrostoma miyazakiense 1 0 1 0 0 0 0 0 2 0.4 1 – 2 TNB Family Molineidae Molineus legerae 0 1 0 0 1 3 1 0 6 1.1 1 – 40 TNB, INM, KWB

Trematoda Family Echinostomatidae Isthmiophora hortensis 0 0 1 2 4 4 2 13 26 4.9 1 – 101 TNB, INM, KWB Unidentified echinostomatid 2 1 0 0 1 2 0 3 9 1.7 1 – >200 TNB, INM, KWB sp. with 34–39 collar spines Family Heterophyidae Metagonimus takahashii 1 0 2 1 8 19 7 28 66 12.4 1 – ca 3,080 TNB, HKG, KTD, INM, KWB, GBO, YRA Metagonimus yokogawai 0 0 0 0 1 2 0 1 4 0.8 3 – >173 TNB, OHT, HKG, GBO Family Plagiorchiidae Plagiorchis muris 010000001 0.21 KWB Family Nanophyetidae Macroorchis spinulosus 2 0 1 4 0 3 0 0 10 1.9 1 – 41 TNB, OHT, HKG, KWB Family Dicrocoeliidae Concinium ten 000100001 0.24 TNB

Cestoda Family Mesocestoidae Mesocestoides sp. 0 0 0 0 1 0 0 0 1 0.2 8 TNB

Acanthocephala Family Centrorhynchidae Centrorhynchus bazaleticus 000000011 0.21 TNB Centrorhynchus teres 0 0 2 1 6 12 0 8 29 5.5 1–17 TNB, OHT, KTD, KWB, GBO, YRA Sphaerirostris lanceoides 0 0 0 1 2 6 0 4 13 2.4 1 TNB, HSM Family Plagiorhynchidae Plagiorhynchus ogatai 0 0 1 0 1 1 0 0 3 0.6 1 TNB, INM Porrorchis oti 0 0 0 0 1 6 0 1 8 1.5 1 TNB, OHT, INM Family Polymorphidae Southwelina hispida 0 0 0 0 1 3 0 6 10 1.9 1 TNB, OHT, YRA * For abbreviation of the locality, see the legend of Fig. 1. ** Recovery of S. procyonis began from May, 2004, and prevalence was caliculated for the second-year period only (total number of examined raccoons were 286 for this instance). 314 H. SATO AND K. SUZUKI

phasmids situated at around the middle between the second and third pairs of posteroventral papillae (Fig. 4E). Spicules were unequal in length; average length (range, n=4) of left spicule was 0.74 (0.62–0.83) mm, and that of right spicule was 0.65 (0.50–0.77) mm. Contracaecum spiculigerum (Rudolphi, 1809) Railliet et Henry, 1912: Two male and 1 female adults were firmly attached to the gastric mucosa of one raccoon (Fig. 5). In addition, a single juvenile worm each was collected from the intestine of 4 raccoons. These 5 raccoons were trapped in certain areas (Inari, Nakahaya, and Maro) of Tanabe. Mor- phological data of 2 adult males were as follows (in mm): body, 30.0 and 28.0 long, 0.92 and 0.80 wide; head, 0.10 and 0.08 long, 0.23 and 0.20 wide; interlabia 0.074 and Fig. 3. Gross appearance of Physaloptera sp. found in the stom- 0.076 long; nerve ring and cervical papilla from the anterior ach. The four smaller adults on the left side are males with end, 0.56 and 0.48, and 0.66 and 0.61, respectively; esopha- swollen caudal ends, while the other four on the right side are gus, 3.5 and 3.0 long, ventriculus, 0.29 by 0.21 and 0.24 by females. The more tapered end of each worm is the anterior 0.20; ventricular appendix, 1.00 and 0.76 long; intestinal end. cecum, 2.6 and 2.4 long; spicules alate, subequal, 8.94 and 7.85 long; preanal papillae, 46 and 52 pairs; postanal papil- larly at certain foci such as Shinjo and Nakahaya. Figure 4 lae, subventral 4 pairs with lateral 3 pairs; and tail, 0.22 and shows the morphological details of this species. Three pre- 0.20 long. The single female was 17.8 by 0.46, and not fully cloacal sessile papillae of equal size were arranged in a tri- developed without egg production. The juvenile forms angular row, and the external surface of the cloaca was not measured 3.0–4.8 by 0.10–0.15 mm. clearly divided from the surrounding area (Fig. 4D). The Echinostomatid trematodes: The echinostomatid species first pair of posteroventral papillae was situated distantly with 34–39 collar spines were not fully-developed at any from 2 pairs of postcloacal papillae (Fig. 4C). One pair of instances, and DNA extraction from any lots of the parasite

Fig. 4. High magnification of Physaloptera sp. A and B. Different views of the anterior end. Two each of cervical papillae (arrow heads) located on each pseudolabium. C. Ventral view of the caudal end of a male to illustrate the posteroventral microtopography. Upper and lower insets in C are magnified in D and E, respectively. Abbrevia- tion: Cl, cloaca; EP, excretory pore; NR, nerve ring; Ph, phasmid; PostCP, postcloacal papillae; PP, pedunculated papillae; PreCP, precloacal sessile papillae; PVP, posteroventral papillae; PVP-1, the first pair of PVP; PVP-2, the second pair of PVP; and PVP-3, the third pair of PVP. Magnification of A - C is identical. HELMINTHS OF FERAL RACCOONS IN JAPAN 315

Fig. 6. Contracted (left side) and expanded shapes (right side) of Fig. 5. Adult (A, B) and juvenile (C) Contracaecum spicu- Macroorchis spinulosus. OS, oral sucker; Ph, pharynx; SV, ligerum found in the stomach and intestine, respectively. seminal vesicle; Te, testis; VS, ventral sucker; and Vt, vitelline To show clearly the relationship of each organ, the out- gland. line is traced partially. Abbreviations: CP, cervical papilla; Es, esophagus; IC, intestinal cecum; IL, interla- Since other helminth species collected from feral raccoons bia; Int, intestine; NR, nerve ring; Pa, papilla; VA, ven- in the present study are well-known parasites of indigenous tricular appendix; and Ve, ventriculus. wild carnivores in Japan [39, 40], taxonomic note for each parasite is omitted in the present study. was unsuccessful, so that we could not identify the species. Fully-developed Isthmiophora hortensis (Asada, 1926) DISCUSSION Kostadinova et Gibson, 2002, having 27 or 28 collar spines and eggs measuring 0.110–0.136 mm by 0.060–0.076 mm, Among many zoonotic pathogens transmittable through were collected from 26 raccoons. Two testes were contigu- raccoons, the raccoon roundworm (B. procyonis) has the ous, and anterior testis, oval in shape, was equatorial, not highest priority with respect to public health and ecological pre-equatorial. Three arbitrarily chosen specimens from concerns [11, 17, 18, 38, 41, 44, 47]. In this sense, the fact different raccoons had an identical 261-bp COI segment and that no roundworms were found in all 531 raccoons exam- 6,871-bp rDNA sequences (DDBJ/EMBL/GenBank acces- ined in the present study is good news for the residents in the sion nos. AB189980 and AB189982). ITS-1 (444 bp in survey area. However, of concern was the detection of S. length) and ITS-2 (440 bp) of our specimens showed 99% procyonis at a high rate widely in the survey area, since this homology with those of I. hortensis (syn. Echinostoma species is known to cause “creeping eruption” and intestinal hortense; accession no. U58101), and 92% or 94% homol- infection in a healthy human volunteer [21–23]. Detailed ogy with respective regions of I. melis (syn. Euparyphium discussion about this exotic species was made in a separate melis; AY168932). The data of COI and ND1 sequences work [42]. also supported that our specimens should be classified as I. As we reported here, feral raccoons in Japan had meager hortensis. parasites with regard to species number and infection inten- Macroorchis spinulosus Ando, 1918: This minute trema- sity [4, 53], compared with that found in North America tode (≤0.92 mm in length; Fig. 6), numbered 1 to 41 where raccoons are endemic. Most of the collected parasites (geomean, 3.6)/host, was found in 10 raccoons: 4 in Tanabe, from feral raccoons in Japan were food-borne, indigenous 4 in Hikigawa, and 1 each in Ohtoh and Kawabe. helminth species, and infection with soil-transmitted nema- Cestodes were found only in a single raccoon. Found todes was rare in them. It has been reported that the typical were 8 destrobilated adults of Mesocestoides sp. Detailed parasite fauna of raccoons is largely lost after placing the descriptions of Strongyloides procyonis and acanthoceph- host at a new locality or island separated from inland habi- alan species are made in separate publications [42, 43]. tats, and notable variation exists in the diversity and inten- 316 H. SATO AND K. SUZUKI sity of parasitic infection in raccoons from different from feral raccoons. To clarify the origin of this parasite, geographic regions [7, 13, 15, 33, 36]. Originally, feral rac- extensive parasitological investigation of indigenous carni- coons entered Japan as companion animals: the animals vores in the same survey areas is in progress at present. were imported when they were juvenile, approximately 2–3 Molecular phylogenetic relationships and taxonomic month old. Accordingly, imported raccoons might have no revision of abundant echinostomatid spp. (more than 120 time to acquire a variety of parasites before arrival to Japan, nominal spp.) are currently the focus of researchers [19, 20, and in an artificial environment during personal keeping, 28–30]. Apparently, Isthmiophora hortensis detected in this they could not sustain the life cycle of parasites that need study resembled closely I. beaveri (Yamaguti, 1958) Kosta- intermediate hosts. Even if raccoons harboring parasitic dinova et Gibson, 2002, which is the North American sib- helminths are released in the field, it might be again difficult ling species of the Euro-Asian I. melis (Schrank, 1788) to sustain the life cycle of the parasite by a limited number Lühe, 1909, and different from the latter species in shape of dispersing hosts because most species need a specific and position of the testes [5, 12, 19]. Asakawa et al. [4] also intermediate host and the contact between the potential found a closely related species to one described here, in feral intermediate host and final host is fully accidental. After all, raccoons in Hokkaido, Japan, although they reported that transplanted hosts are deprived of the original parasite the two testes are absolutely post-equatorial. In the rede- fauna, and acquire new parasites sustained by indigenous scription of I. hortensis, Kostadinova and Gibson [19] char- hosts in that ecosystem. Most of helminth species collected acterized the position of the testes in either I. melis or I. from feral raccoons in Japan by other researchers [4, 53] and hortensis as pre-equatorial for the anterior testis and equato- us also follow this schema. Stuewer [48] found that adult rial for the posterior testis. Despite the morphological dis- and juvenile male raccoons in Michigan had larger home crepancy from I. hortensis, the nucleotide sequences of ranges than females of corresponding ages, and that adults rDNA did indicate that the species with 27 or 28 collar had larger home ranges than juveniles. This behavioral dif- spines found in the present study was I. hortensis. It is nec- ference and acting power to live food resources might par- essary to consider that greater morphological diversity of tially explain different prevalences by sex noted in the this species might occur in specimens from different host present study (17.5–38.1 % in females vs. 28.6–52.6% in species, or artificially in specimens recovered from frozen males). carcasses. Alternatively, it could be a local variant in Two Physaloptera spp., Physaloptera rara and P. maxil- Wakayama Prefecture. Species differentiation of echinosto- laris, have been recorded at a high prevalence (63–97%) in matids based only on morphology is sometimes difficult, raccoons of North America [9, 35, 46]. The former species and nucleotide sequencings may provide a solution to this is a common stomach parasite of coyotes (Canis latrans problem [28]. Say, 1823), while the latter is a common stomach parasite of Macroorchis spinulosus was originally described by skunk (Mephitis mephitis (Schreber, 1776)). Anderson [1] Ando [2, 3] in 1918 by experimental infection of laboratory pointed out that P. maxillaris could not infect raccoons animals with metacercariae encysted in Japanese freshwater experimentally, and the reports of P. rara from raccoons crabs (Geothelphusa dehaani) collected in Gifu Prefecture, might need confirmation as well. In Japan, P. praeputialis, near our survey area. Although a variety of laboratory ani- a common stomach worm of felids with cosmopolitan distri- mals including dogs, cats, mice, rats, guinea-pigs, and rab- bution, has been recorded in domestic cats and dogs [14, bits served as the experimental definitive hosts, the natural 30], but any Physaloptera species has never been recorded definitive host was unknown until 1982, when Saito et al. in wild carnivores. P. praeputialis is distinguished from P. [37] found the natural infection in Japanese water shrews rara by several morphological features [50]. In the poster- (Chimarrogale platycephala platycephala (Temminck, oventral microtopography except for the position of the 1842)) collected in Shizuoka Prefecture with adult flukes, phasmids (Fig. 4), and the length of spicules of our speci- and in Japanese freshwater crabs collected in Shizuoka, mens (left, 0.62–0.83 mm; and right, 0.50–0.77 mm) and P. Kochi, and Nagasaki Prefectures with metacercariae. They rara (left, 0.74–0.90 mm; and right, 0.48–0.70 mm [32]), found also two new species of the Macroorchis (i.e., M. chi- these two species resembled closely each other, distinct marrogalus and M. elongatus) in Japanese water shrews from P. praeputialis. P. sibirica is known from foxes [37], and Machida and Uchida [26] recorded another new (Vulpes vulpes (Linnaeus, 1758)), badgers (Meles meles species, M. himizu, from greater Japanese shrew-moles (Linnaeus, 1758)), dogs, and cats at cold climates or high (Urotrichus talpoides Temminck, 1841). As shown in Fig. altitudes of the Eurasian continent [25, 34, 45, 51]. The 6, our specimens are identical to the original description of length of spicules of P. sibirica (left, 0.59–0.74 mm; and M. spinulosus made by Ando [2, 3]. Recently, Korean right, 0.53–0.64 mm) [45], and the posteroventral microto- researchers redescribed this species based on distinct spe- pography except for the arrangement of three precloacal cies, which was obtained experimentally in mice, rats and sessile papillae (a flattened triangular row in our specimens cats by inoculating metacercariae encysted in Korean fresh- vs. a distinct triangular row in P. sibirica [25, 34]) are close water crayfishes (Cambaroides similis) from the eastern to our speciemens. The position of the posteroventral phas- mountainous areas of Korea [6]. This Korean species is mids of P. sibirica is identical with our specimens. Accord- closely related to M. himizu, but presumably a distinct spe- ingly, we could not specify Physaloptera species obtained cies. HELMINTHS OF FERAL RACCOONS IN JAPAN 317

Adults and juveniles of Contracaecum spiculigerum were 8. Chisholm, L. A., Morgan, J. A. T., Adlard, R. D. and Whitting- isolated from 5 raccoons caught in the same area: 3 devel- ton, I. D. 2001. Phylogenetic analysis of the monocotylidae oped adults attached to the stomach of one raccoon, and one (Monogenea) inferred from 28S rDNA sequences. Int. J. Para- juvenile each was found in the intestine of 4 raccoons. The sitol. 31: 1253–1263. localities of parasite collection are near the nest area of com- 9. Cole, R. A. and Shoop, W. L. 1987. Helminths of the raccoon (Procyon lotor) in western Kentucky. J. Parasitol. 73: 762– mon cormorants (Phalacrocorax carbo (Linnaeus, 1758)), 768. although the population of this avian host has decreased (ca. 10. Furuoka, H., Sato, H., Kubo, M., Owaki, S., Kobayashi, Y., 2,000 in 1992 to ca. 500 in 2001; M. Tsumura, unpublished) Matsui, T. and Kamiya, H. 2003. Neuropathological observa- after dispersion of raccoons around there. tion of rabbits (Oryctolagus cuniculus) affected with raccoon Raccoons favor not only rural or suburban life but also roundworm (Baylisascaris procyonis) larva migrans in Japan. urban life. When we consider the wandering of this carni- J. Vet. Med. Sci. 65: 695–699. vore within the town or from the wild to the town, or vice 11. Gavin, P. J., Kazacos, K. R., Tan, T. Q., Brinkman, W. B., versa, feral raccoons make a special situation with respect to Byrd, S. E., Davis, A. D., Mets, M. B. and Shulman, S. T. dispersion of viral, bacterial, protozoan and helminthic dis- 2002. Neural larva migrans caused by the raccoon roundworm eases to residents, companion or domestic animals, and the Baylisascaris procyonis. Pediatr. Inf. Dis. J. 21: 971–975. 12. Gupta, S. P. 1962. A redscription of Euparyphium melis indigenous wildlife, along with their wandering. In this (Schrank, 1788) Dietz 1909 and Echinostoma revolutum context, it should be stressed that feral raccoons must be (Froelich, 1802) Looss, 1899 parasitic in the intestine of mink controlled or eradicated promptly not only for their negative (Mustela vison) and muskrat (Ondatra zibethica) from Canada. economic impact through the destruction of gardens and Ind. J. Helminthol. 14: 77–85. orchards, but also from the viewpoint of disease control for 13. Harkema, R. and Miller, G. C. 1962. Helminths of Procyon the public, domestic animals and wildlife. Even if this is lotor solutus from Cape Island, South Carolina. J. Parasitol. impossible, exotic raccoons should be routinely surveyed 48: 333–335. for their zoonotic or exotic pathogens. 14. Hayasaki, M., Oishi, I. and Munakata, A. 1982. Incidence of stomach worm, Physaloptera praeputialis von Linstow, 1889, ACKNOWLEDGMENTS. We express our sincere thanks in two cats and a dog in Tokyo, Japan. Jpn. J. Parasitol. 31: 499–506. to trappers and officers engaged in control measures of feral 15. Hoberg, E. P. and McGee, S. G. 1982. Helminth parasitism in raccoons in Wakayama Prefecture who made this study pos- raccoons, Procyon lotor hirtus Nelson and Goldman, in sible, Dr. M. Horiuchi, Hokkaido University, Miss Y. Naya, Saskatchewan. Can. J. Zool. 60: 53–57. Wildlife Management Office, Kawasaki, and Dr. Y. Yoko- 16. Johnson, A. S. 1970. Biology of the raccoon (Procyon lotor hata, Toyama University, for their kind arrangement of our varius) Nelson and Goldman in Alabama. Auburn Univ. Agr. collaborative study, and Wakayama prefectural office for Exp. St. Bull. 402: 1–148. the permission to use the unpublished inquiry results. This 17. Kazacos, K. R. 1997. Visceral, ocular, and neural larva study was supported partly by a Grant-in-Aid (No. migrans. pp. 1459–1473. In: Pathology of Infectious Diseases, 13460137) from the Japan Society for the Promotion of Sci- vol. II (Coonor, D. H., Chandler, F. W., Schwartz, D. A., ence. Manz, H. J. and Lack, E. E. eds.), Appleton, Stamford, Con- necticut. 18. Kazacos, K. R. 2001. Baylisascaris procyonis and related spe- REFERENCES cies. pp. 301–341. In: Parasitic Diseases of Wild Mammals, 2nd ed. (Samuel, W. M., Pybus, M. J. and Kocan, A. A. eds.), 1. Anderson, R. C. 1992. Nematode Parasites of Vertebrates; Iowa State University Press, Ames, Iowa. Their Development and Transmission. C.A.B. International, 19. Kostadinova, A. and Gibson, D. I. 2002. Isthmiophora Lühe, Oxon. 1909 and Euparyphium Dietz, 1909 (Digenea: Echinostoma- 2. Ando, A. 1918. On a new tematode with a freshwater crab as tidae) re-defined, with comments on their nominal species. its intermediate host. Tokyo Iji Shinshi 2079: 1215–1220 (in Syst. Parasitol. 52: 205–217. Japanese). 20. Kostadinova, A., Herniou, E. A., Barrett, J. and Gibson, D. I. 3. Ando, A. 1918. On a new tematode with a freshwater crab as 2003. Phylogenetic relationships of Echinostoma Rudolphi, its intermediate host. Tokyo Iji Shinshi 2081: 1320–1327 (in 1809 (Digenea: Echinostomatidae) and related genera re- Japanese). assessed via DNA and morphological analyses. Syst. Parasitol. 4. Asakawa, M., Kurachi, T. and Wildl. Eco. Soc. 1999. Parasitic 54: 159–176. helminths of raccoons in Hokkaido, Japan. Jpn. J. Zoo Wildl. 21. Little, M. D. 1961. Comparative morphology of six previously Med. 4: 101–103 (in Japanese with English abstract). described and seven new species of Strongyloides (Nematoda). 5. Beaver, P. C. 1941. Studies on the life history of Euparyphium Ph.D. thesis, Tulane University, New Orleans, Louisiana. melis (Trematoda: Echinostomidae). J. Parasitol. 27: 35–44. 22. Little, M. D. 1965. Dermatitis in a human volunteer infected 6. Chai, J.-Y., Sohn, W.-M., Huh, S., Choi, M.-H., and Lee, S.-H. with Strongyloides of nutria and raccoon. Am. J. Trop. Med. 1996. Redescription of Macroorchis spinulosus Ando, 1918 Hyg. 14: 1007–1009. (Digenea: Nanophyetidae) encysted in the fresh water crayfish, 23. Little, M. D. 1966. Seven new species of Strongyloides (Nema- Cambaroides similis. Korean J. Parasitol. 34: 1–6. toda) from Louisiana. J. Parasitol. 52: 85–97. 7. Ching, H. L., Leighton, B. J. and Stephen, C. 2000. Intestinal 24. Lotze, J.-H. and Anderson, S. 1979. Procyon lotor. Mamm. Sp. parasites of raccoons (Procyon lotor) from southwest British 119: 1–8. Columbia. Can. J. Vet. Res. 64: 107–111. 25. Miquel, J., Segovia, J. M., Feliu, C. and Torres, J. 1996. On 318 H. SATO AND K. SUZUKI

Physaloptera sibirica Petrow et Gorbunow, 1931 (Nematoda: paucitesticulus and Brachylaima tokudai. J. Vet. Med. Sci. 61: Physalopteridae) parasitizing Iberian mammals. Wiad. Parazy- 1339–1342. tol. 42: 435–442. 40. Sato, H., Inaba, T., Ihama, Y. and Kamiya, H. 1999. Parasito- 26. Machida, M. and Uchida, A. 1982. Some helminth parasites of logical survey on wild Carnivora in north-western Tohoku, the Japanese shrew mole from the Izu Peninsula. Mem. Natl. Japan. J. Vet. Med. Sci. 61: 1023–1026. Sci. Mus., Tokyo 15: 149–154 41. Sato, H., Kamiya, H. and Furuoka, H. 2003. Epidemiological 27. Miyashita, M. 1993. Prevalence of Baylisascaris procyonis in aspects of the first outbreak of Baylisascaris procyonis larva raccoons in Japan and experimental infections of the worm in migrans in rabbits in Japan. J. Vet. Med. Sci. 65: 453–457. laboratory animals. Seikatsu-eisei 37: 137–151 (in Japanese 42. Sato, H., Suzuki, K., Osanai, A., Kamiya, H. and Furuoka, H. with English abstract). 2005. Identification and characterization of the threadworm, 28. Morgan, J. A. T. and Blair, D. 1995. Nuclear rDNA ITS Strongyloides procyonis, from feral raccoons (Procyon lotor) sequence variation in the trematode genus Echinostoma: an aid in Japan. J. Parasitol. 92: 63–68. to establishing relationships within the 37-collar-spine group. 43. Sato, H., Suzuki, K., Uni, S. and Kamiya, H. 2005. Recovery Parasitology 111: 609–615. of the everted cystacanth of seven acanthocephalan species of 29. Morgan, J. A. T. and Blair, D. 1998. Relative merits of nuclear birds from feral raccoons (Procyon lotor) in Japan. J. Vet. Med. ribosomal internal transcribed spacers and mitochondrial CO1 Sci. 67: 1203–1206. and ND1 genes for distinguishing among Echinostoma species 44. Sato, H., Une, Y., Kawakami, S., Saito, E., Kamiya, H. and (Trematoda). Parasitology 116: 289–297. Furuoka, H. 2005. Fatal Baylisascaris larva migrans in a col- 30. Morgan, J. A. T. and Blair, D. 1998. Mitochondrial ND1 gene ony of Japanese macaques kept by a safari-style zoo in Japan. sequences used to identify echinostome isolates from Australia J. Parasitol. 91: 716–719. and New Zealand. Int. J. Parasitol. 28: 493–502. 45. Skrjabin, K. I. and Soboleb, A. A. 1964. [Essentials of Nem- 31. Oishi, I. and Kume, S. 1974. On the stomach worm found from atodology, vol. 12 (Skrjabin, K. I. ed.): Spirurata of animals cats in Tokyo. Jpn. J. Vet. Sci. 36: 27–32. and man and the diseases caused by them; 2nd part, Physalop- 32. Olsen, O. W. 1974. Animal Parasites: Their Life Cycles and teridae], Izdat. Akad. Nauk, SSSR, Moscow (in Russian). Ecology, 3rd ed., University Park Press, Baltimore. 46. Smith, R. A., Kennedy, M. L. and Wilhelm, W. E. 1985. Helm- 33. Price, R. L. and Harman, D. M. 1983. Helminths from the rac- inth parasites of the raccoon (Procyon lotor) from Tennessee coon, Procyon lotor litoreus Nelson and Goldman 1930, on St. and Kentucky. J. Parasitol. 71: 599–603. Catherines Island, Georgia. Proc. Helminthol. Soc. Wash. 50: 47. Snyder, D. E. 1988. Indirect immunofluorescent detection of 343–344. oocysts of Cryptosporidium parvum in the feces of naturally 34. Quentin, J.-C. and Biocca, E. 1976. Presence du Nématode infected raccoons (Procyon lotor). J. Parasitol. 74: 1050– Physaloptera sibirica Petrow et Gorbunow, 1931, parasite de 1052. carnivores chez le lerot Eliomys quercinus L. dans les Alpes. 48. Stuewer, F. W. 1943. Raccoons: their habits and management Ann. Parasitol. (Paris) 51: 255–262 (in French). in Michigan. Ecol. Monogr. 13: 203–257. 35. Richardson, D. J., Owen, W. B. and Snyder, D. E. 1992. Helm- 49. Thompson, J. D., Higgins, D. G. and Gibson, T. J. 1994. inth parasites of the raccoon (Procyon lotor) from north-central CLUSTRAL W: Improving the sensitivity of progressive mul- Arkansas. J. Parasitol. 78: 163–166. tiple sequence alignment through sequence weighting, posi- 36. Sadykhov, I. A. 1966. [The helminth fauna of raccoons accli- tions-specific gap penalties and weight matrix choice. Nucleic matized in Azerbaidzhan. p. 246. In: Acclimatization of Ani- Acids Res. 22: 4673–4680. mals in the U.S.S.R. (Yanushevich, A. I. ed.), translated from 50. Tiekotter, K. L. 1981. Observations of the head and tail regions Russian by the Israel Program for Scientific Translations, of male Physaloptera praeputialis von Linstow, 1889, and Jerusalem. Physaloptera rara Hall and Wigdor, 1918, using scanning 37. Saito, Y., Watanabe, T. and Yamashita, T. 1982. Two new electron microscopy. Proc. Helminthol. Soc. Wash. 48: 130– intestinal trematodes, Macroorchis chimarrogalus n. sp. and 136. Macroorchis elongatus n. sp. from Japanese water shrew and 51. Torres, J., Miquel, J. and Motjé, M. 2001. Helminth parasites others, with a description of the metacercaria of Macroorchis of the Eurasian badger (Meles meles L.) in Spain: a biogeo- chimarrogalus n. sp. (Trematoda: Nanophyetidae). Acta Med. graphic approach. Parasitol Res. 87: 259–263. Biol. 30: 47–56. 52. Van de Peer, Y., de Rijk, P., Wuyts, J., Winkelmans, T. and de 38. Sato, H., Furuoka, H. and Kamiya, H. 2002. First outbreak of Wachter, R. 2000. The European small subunit ribosomal RNA Baylisascaris procyonis larva migrans in rabbits in Japan. Par- database. Nucleic Acids Res. 28: 175–176. asitol. Int. 51: 105–108. 53. Yamada, D., Oku, Y., Nonaka, N., Asakawa, M., Ikeda, T., 39. Sato, H., Ihama, Y., Inaba, T., Yagisawa, M. and Kamiya, H. Asano, M., Akamatsu, R., Matoba, Y. and Kamiya, M. 2000. 1999. Helminth fauna of carnivores distributed in north-west- Studies on the parasite fauna of raccoon (Procyon lotor) natu- ern Tohoku, Japan, with special reference to Mesocestoides ralized in Hokkaido, Japan. Parasitol. Int. 49 (Suppl.): 91.