Trichinella nativa and T. spiralis induce distinguishable histopathologic and humoral responses in the raccoon dog (Nyctereutes procyonoides)

Three experimental groups of six male raccoon dogs (Nyctereutes procyonoides) each were formed by placing one of three littermates from six litters into each group. One group was inoculated with pig-origin Trichinella spiralis, the second was inoculated with raccoon dog-origin T. nativa, and the third served as a control group. The infective dose was 1,000 larvae/kg of body weight. Every third week, biopsies from M. triceps brachii were taken, and serum samples were collected for up to 12 weeks postinfection. In the early phase of the infection, cysts of both parasites were elongated cylinders that later became more spherical. However, at the end of the experiment, the cysts of T. nativa were more rounded than those of T. spiralis (mean length/width = 2.5 versus 1.5 in T. spiralis versus T. nativa, respectively). Both species accumulated a collagen-rich capsule around the nurse cell, but the capsule was thicker in T. nativa. In both parasites, the total surface area of the sagittal section of the cyst was equal. Inflammation was more intense around T. nativa cysts. Specific antibodies were recognizable 2 weeks after infection by both enzyme-linked immunosorbent assay (ELISA) and western blot. In western blots, serum from both T. nativa- and T. spiralis-infected animals recognized the same components, but reaction with the homologous antigen was stronger. The same pattern was also seen in the ELISA. Immunoreactive epitopes were localized only in internal organs and cuticula of larvae in muscle.     

Histopathological comparison of pulmonary artery lesions between raccoon dogs (Nyctereutes procyonoides) and domestic dogs experimentally infected with Dirofilaria immitis

Five raccoon dogs (Nyctereutes procyonoides) and two domestic dogs (Canis familiaris) were subcutaneously infected with 100 infective larvae (L3) of Dirofilaria immitis. Two and five worms, respectively, were collected from two of three raccoon dogs. Villous endarteritis was found in the raccoon dog with five worms and two dogs at 116 days after infection. The number of recovered worms in the raccoon dogs was significantly smaller than that of the domestic dogs having 22 and 29 worms, while histopathological features and the severity of the lesions in the raccoon dogs were similar to those in the domestic dogs. The vascular lesions in two chronically-infected raccoon dogs turned into much severe at 565 and 590 days after inoculation.     

Sarcoptic mange in free-ranging raccoon dogs (Nyctereutes procyonoides) in Japan

Sarcoptes scabiei infestation was diagnosed in three freshly dead free-ranging raccoon dogs (Nyctereutes procyonoides) in Kanagawa Prefecture, Japan. The dogs presented with an alopecic pruritic skin disease, with signs of alopecia on the ears, muzzle, around the eyes, elbow, thigh and the neck, and hyperpigmented and crusted skin lesions, which had a severe malodour. Skin scrapings revealed the presence of the mite Sarcoptes scabiei. Histopathology of lesions demonstrated marked acanthosis, hyperkeratosis, parakeratosis and fungal elements, which were subsequently identified as Acremonium sp., Alternaria sp. and an unknown fungus. Mite segments were located mainly in the stratum corneum and also in the stratum granulosum. Tunnels could be observed in the hyperkeratotic stratum corneum. Scanning electron microscopy (SEM) revealed the tortoise-like Sarcoptes scabiei with four long bristles, suckers and blade-like claws on legs 1 and 2, cuticular spines, prominent body striations and a terminal anus. SEM also revealed an adult female mite digging a tunnel with the head wedged into the very end of the closed burrow. Tunnels filled with eggshells, corneocyte debris and faecal pellets were also observed.     

Helminth findings in indigenous raccoon dogs Nyctereutes procyonoides (Gray, 1843)

Internal organs of 74 racoon dogs originating from the eastern districts of the Federal Land Brandenburg were subjected to a complete helminthological dissection. In addition, samples of diaphragma muscles from 46 further animals were examined for Trichinella larvae. Altogether 3 trematode- (Alaria alata, Isthmiophora melis, Metorchis bilis), 3 cestode- (Mesocestoides sp., Echinococcus multilocularis, Taenia polyacantha) and 7 nematode species (Capillaria aerophila, C. plica, Trichinella spiralis, Uncinaria stenocephala, Molineus patens, Crenosoma vulpis, Toxocara canis) could be identified. Thereby, the racoon dog has a comparable helminth spectrum as well known in red foxes. Differences in the frequency of the occurrence of helminths in both hosts can be explained by different food preferences.     

Radiographic evaluation of congenital vertebral anomalies in Korean raccoon dogs (Nyctereutes procyonoides koreensis)

BackgroundThe normal vertebral anatomy of Korean raccoon dogs and their variants require research attention as a prerequisite for identifying pathologies and anomalies.ObjectivesThis retrospective study aimed at describing the vertebral formula and congenital vertebral anomalies in Korean raccoon dogs (Nyctereutes procyonoides koreensis).MethodsRadiographs of 82 raccoon dogs (42 males, 40 females) acquired from May 2013 to June 2020 in the Gangwon Wildlife Medical Rescue Center were reviewed to evaluate the cervical, thoracic, and lumbar vertebrae of the spine.ResultsNormal morphology of all vertebrae was observed in 50 of the 82 raccoon dogs, and the vertebral formula was cervical 7, thoracic 13, and lumbar 7. Congenital vertebral anomalies were found in 32 raccoon dogs: transitional vertebrae (TV) in 31 and block vertebrae in 2. Two raccoon dogs had 2 types of vertebral anomalies: one had TV and block vertebra, and the other had 2 types of TV. Twenty-nine raccoon dogs had thoracolumbar TV (TTV) and 3 had lumbosacral TV. TTV was morphologically classified into 4 different types: unilateral extra-rib in 5 raccoon dogs, bilateral extra-ribs in 14, bilateral elongated transverse processes in 4, and an asymmetric mixed formation of extra-rib with elongated transverse process in 6.ConclusionsThis study showed that TTV is common in Korean raccoon dogs, and that the vertebral formula is relatively diverse. The bilateral extra-ribs type TTV is the most common variant, which is almost similar to normal rib to be confused the radiographic evaluation.     

Severe hydrocephalus in a raccoon dog (Nyctereutes procyonoides)

Hydrocephalus is one of the most common central nervous system malformations in domestic dogs, yet they are poorly documented and studied in wild carnivoran mammals. A pup of raccoon dog (Nyctereutes procyonoides) was rescued and brought to Wildlife Center. The pup showed generalized ataxia, a domed skull, and an open bregmatic fontanelle. Ultrasound and MRI showed severe enlargement of the lateral ventricle with the loss of septum pellucidum resulting in a single large ventricle and cervical syringohydromyelia. Although treatment was attempted, the animal was euthanized due to poor prognosis. At necropsy, macroscopic findings were identical to the diagnostic imaging, where marked enlargement of the calvarium, and attenuated gyri and sulci were observed. Finally, hydrocephalus was confirmed. Here, we describe a case of hydrocephalus in a raccoon dog (Nyctereutes procyonoides).     

Epidemiology of sarcoptic mange in free-ranging raccoon dogs (Nyctereutes procyonoides) in Yokohama,Japan

Free-ranging raccoon dogs (Nyctereutes procyonoides) from Nogeyama Zoological Gardens, Kanazawa Zoological Gardens, and Yokohama Zoological Gardens frequently rescued dogs having Sarcoptes scabiei infestation. However, the epidemiology of S. scabiei infestation has not yet been elucidated. In the present study, we investigated the epidemiology of S. scabiei infestation in raccoon dogs and its influence on the population of masked palm civets in Yokohama, Japan. We examined records of raccoon dog rescue between 1981 and 2010 and classified the dogs into the following 4 categories on the basis of the reason for rescue: dogs with S. scabiei infestation, scabies-infested dogs involved in car accidents, uninfested dogs involved in car accidents, and other reasons for rescue. We found that the number of dogs rescued due to car accidents and other reasons increased from 1989 onwards, and an S. scabiei outbreak was recorded since 1993. The infestation spread from the southern to the northern regions of Yokohama. The total number of raccoon dogs rescued annually peaked in 1995 and declined thereafter. The number of masked palm civets (Paguma larvata) rescued gradually increased with a decline in the number of raccoon dogs rescued. In the present study, we revealed the epidemiology of S. scabiei infestation in the raccoon dog. The outbreak might be induced by the increased population density, and the infestation spread immediately from the southern to the northern regions of Yokohama since 1993. Further, the population of masked palm civets may have increased due to the decrease in the population of the raccoon dog.     

Epidemiology of canine distemper virus in wild raccoon dogs (Nyctereutes procyonoides) from South Korea

Raccoon dogs (Nyctereutes procyonoides) are widespread and common in South Korea. In 2011, we obtained serum samples from 102 wild raccoon dogs to survey their exposure to canine distemper virus (CDV). Forty-five of the 102 animals (44.1%) were seropositive. Field cases of canine distemper in wild raccoon dogs from 2010 to 2011 were investigated. Fourteen cases of CDV infection were identified by a commercially available CDV antigen detection kit. These cases were used for virus isolation and molecular analysis. Sequence analysis of hemagglutinin genes indicated that all viruses isolated belonged to the Asia-2 genotype. H protein residues which are related to the receptor and host specificity (residues 530 and 549) were analyzed. A glutamic acid (E) residue is present at 530 in all isolates. At 549, a histidine (H) residue was found in five isolates and tyrosine (Y) residue was found in 6 isolates. Our study demonstrated that CDV infection was widespread in wild raccoon dogs in South Korea.     

Detection of antibodies against the rabies virus in Korean raccoon dogs (Nyctereutes procyonoides koreensis)

Rabies has been reported in dogs, cattle, and raccoon dogs in Korea, especially in the provinces of Gyeonggi and Gangwon. The aim of this study was to investigate the seroprevalence of the rabies in wild Korean raccoon dogs by using 50 blood samples collected in Gyeonggi and Gangwon provinces. Antibodies against rabies were measured by using a fluorescent antibody virus neutralization test. Of the 50 blood samples tested, 20 raccoon dogs (40%) tested positive. Seropositive animals were distributed in Paju, Yangpyeong, Goyang, Yangju, Yanggu, and Hoengseong cities.     

Nematodes from raccoon dogs (Nyctereutes procyonoides viverrinus) introduced recently on Yakushima Island, Japan

Nematodes were collected from 14 raccoon dogs (Nyctereutes procyonoides viverrinus) on Yakushima Island, Kagoshima Prefecture (Kyushu), Japan, that were the progeny of recently introduced animals to this World Natural Heritage Area, probably within the last 2 decades. Seven nematode species (Toxocara tanuki, Ancylostoma kusimaense, Arthrostoma miyazakiense, Strongyloides planiceps, Trichuris vulpis, Gongylonema sp., and Dirofilaria immitis) were collected from the raccoon dogs. Two nematode species (Molineus legerae and Strongyloides martis) were collected from 2 Japanese weasels (Mustela itatsi) indigenous to this island that were killed by traffic accidents. Considering that foxes and other carnivores, except for weasels, dogs and cats, are not distributed on this island, most of soil-borne nematodes such as roundworms, hookworms, and threadworms in raccoon dogs should have been introduced by the original population from the mainland Japan, and the infection hence maintained at a high level by new generations. Recovery of the third-stage larvae of Gongylonema sp. from the esophagus of raccoon dogs was remarkably frequent (79%), suggesting that the animals might vigorously take insect hosts. Although S. martis have never been recorded from weasels in Japan until this study, this threadworm might be prevalent in a variety of mustelids in Eurasia.     

Listeriosis in a raccoon dog (Nyctereutes procyonoides) associated with canine distemper

A wild raccoon dog (Nyctereutes procyonoides) that manifested severe illness and died was examined. Necropsy revealed severe emaciation, systemic icterus and petechial hemorrhages on the mucous membranes. Histopathologically, necroses were seen in the liver and brain stem associated with meningitis. Eosinophilic intranuclear inclusion bodies were observed in the spleen and intestinal mucosa, and eosinophilic intracytoplasmic inclusion bodies were seen in transitional epithelium in the bladder. Listeria monocytogenes 4b was isolated from the liver, spleen, kidneys and lungs, and the pathogen was also detected in the liver and brain stem immunohistopathologically. The disease was diagnosed as listeriosis associated with canine distemper virus infection in a raccoon dog.     

Effects of alprazolam on capture stress-related serum cortisol responses in Korean raccoon dogs (Nyctereutes procyonoides koreensis)

The purpose of this study was to evaluate the effect of alprazolam on the stress that Korean raccoon dogs (Nyctereutes procyonoides koreensis) may experience while caught in a live trap by measuring their serum cortisol response. The animals were placed in a live trap with or without being pretreated with oral doses of alprazolam. In both groups, pre-trap blood samples were initially collected without anesthesia before the animals were positioned in the live trap; then post-trap blood samples were collected after the animals had remained in the live trap for 2 h. Changes in cortisol levels were observed using a chemiluminescent immunoassay. The level of cortisol increased in the control group and decreased in the alprazolam-pretreatment group (p < 0.05). In this study, we demonstrated that alprazolam pretreatment reduced stress during live trap capture.     

Juvenile bird acanthocephalans recovered incidentally from raccoon dogs (Nyctereutes procyonoides viverrinus) on Yakushima Island, Japan

Helminth parasites were collected from 14 raccoon dogs (Nyctereutes procyonoides viverrinus) and 2 Japanese weasels (Mustela itatsi sho) on Yakushima Island, Kagoshima Prefecture (Kyushu), Japan. In addition to trematodes and nematodes reported separately, everted cystacanths of 3 bird acanthocephalans (Mediorhynchus robustus in 5 animals, Porrorchis leibyi in 2 animals, and Sphaerirostris lanceoides in 1 animal) were collected from the raccoon dogs introduced recently on this World Natural Heritage Area. A cystacanth of P. leibyi was collected also from a weasel indigenous to this island that were killed by traffic accidents. Recovery of everted cystacanths of M. robustus from the intestine of raccoon dogs was remarkably frequent (36%), suggesting that this acanthocephalan should be highly prevalent in passerine hosts on Yakushima Island, and the animals might vigorously take insect hosts or paratenic hosts. Furthermore, the present report is the second on M. robustus after its first record in Japan (Aichi Prefecture, Honshu) as M. garruli from a Eurasian jay (Garrulus glandarius japonicus) by Yamaguti in 1939. Porrorchis leibyi has never been recorded in Japan.     

Clinical features of experimental trichinellosis in the raccoon dog (Nyctereutes procyonoides)

Three groups of six raccoon dogs (Nyctereutes procyonoides) were provided for the experiment: the first group was infected with pig-origin Trichinella spiralis, the second with raccoon dog-origin Trichinella nativa, and the third served as controls. Infection dose for both parasite species was 1000 larvae/kg of body weight, which led to intense final infection. Clinical signs, haematology and serum biochemistry with repeated blood samples were monitored up to 12 weeks post-infection. The most significant findings were a short-term eosinophilia in peripheral blood from the end of the first week post-infection until the end of the third week, loss of weight, and mild anaemia. In the early phase of the infection, the animals had gastrointestinal signs, loss of appetite and diarrhoea. No specific differences in clinical findings could be noticed between the groups infected with T. nativa and T. spiralis. In contrast to the symptoms reported in human outbreaks, fever was not observed in any of the infected animals and serum levels of muscle-specific enzymes did not change. No acute-phase response was observed in the enteral or parental phase of the infection. These findings indicate that because Trichinella spp. are very well adapted to the raccoon dog, it thus, could serve as the most crucial reservoir animal for sylvatic trichinellosis in Finland.     

Pathogenesis and serodiagnosis of experimental Trichinella spiralis spiralis and Trichinella spiralis nativa infections in cattle.

Trichinella spiralis spiralis infections were established in cattle by gavage and by feeding infected musculature in the ration. Trichinae were present in greatest numbers in masseter, tongue and diaphragm. Trichinella spiralis nativa had a low infectivity to cattle although a light infection was established in one cow by a heavy challenge. Cattle had an aversion to eating musculature unless it was camouflaged with molasses. Clinical signs of reluctance to eat and masticate were observed between 10 and 30 days postinfection. Eosinophil counts started to increase at seven days and peaked at about 30 days postinfection. By day 60 eosinophil counts returned to near preinfection levels but in animals examined greater than 90 days postinfection, the counts were variable. Focal lesions of eosinophilic myositis were observed up to about 90 days postinfection. Little cellular reaction was observed surrounding trichinae after muscle invasion and cyst development was completed except for cysts undergoing disintegration. Seroconversion occurred in all cattle examined between 7 and 14 days postinfection. Seroconversion was associated with IgG1 and IgG2 immunoglobulins. Peak levels of antibody occurred between 30 and 60 days. Cattle examined at 182 and 369 days postinfection showed a gradual decrease in antibody levels over time.     

First record of Cryptosporidium infection in a raccoon dog (Nyctereutes procyonoides viverrinus)

Cryptosporidium species have been found in more than 150 species of mammals, but there has been no report in raccoon dogs. Here we found the Cryptosporidium organism in a raccoon dog, Nyctereutes procyonoides viverrinus, and identified this isolate using PCR-based diagnostic methods. Cryptosporidium diagnostic fragments of the 18S ribosomal RNA, Cryptosporidium oocyst wall protein and 70-kDa heat shock protein genes were amplified from the isolate and sequenced to reveal the phylogenetic relationships between it and other Cryptosporidium species or genotypes reported previously. The results showed that the raccoon dog isolate represented the C. parvum cattle genotype which could be a causative agent in human cryptosporidiosis.     

Seasonal changes in immunolocalization of inhibin/activin subunits and testicular activity in wild male raccoon dogs (Nyctereutes procyonoides)

Thirty-four pairs of testes from wild adult raccoon dogs (Nyctereutes procyonoides) were obtained between September 2000 and May 2003. The cellular localization of the inhibin alpha and inhibin/activin (betaA and betaB) subunits in wild raccoon dog testes was investigated. The testicular weight and size and seminiferous tubule diameters were measured. There were marked seasonal variations in testicular weight and size and seminiferous tubule diameters, with values relatively low in September and high in March. Spermatogonia and primary spermatocytes were observed in September, and spermatogonia, spermatocytes, and round spermatids were present in January. All types of spermatogenic cells, including mature spermatozoa, were found in March, indicating that the breeding season is around March in Japan. Thereafter, spermatogonia and degenerating spermatocytes were observed in April. The sections of testes were immunostained by the avidin-biotin-peroxidase complex method (ABC) using polyclonal antisera raised against porcine inhibin alpha, inhibin/activin betaA and inhibin/activin betaB. The inhibin alpha and inhibin/activin (betaA and betaB) subunits were only expressed in Leydig cells in September. On the other hand, the inhibin alpha, betaA, and betaB subunits were observed in Leydig cells and Sertoli cells, but not in germ cells, in March. These results suggest that the testes of wild raccoon dogs have the ability to synthesize inhibins, and the cellular localization of inhibin/activin subunits showed season-related changes in the breeding and non-breeding seasons.     

Differentiation of Trichinella spiralis spiralis and Trichinella spiralis nativa based on resistance to low temperature refrigeration.

A refrigeration technique to differentiate the subspecies, Trichinella spiralis spiralis and T. spiralis nativa is described. Trichinella spiralis spiralis trichinae in musculature do not survive 48 hours post-refrigeration at -32 degrees C while T. spiralis nativa will survive 72 hours and longer at the same temperature.