Detection of ehrlichial infection by PCR in dogs from Yamaguchi and Okinawa Prefectures, Japan

Species-specific nested polymerase chain reaction (PCR) was used to detect the presence of possible canine ehrlichial agents (Ehrlichia canis, E. chaffeensis, E. ewingii, E. equi and E. platys) and monocytic ehrlichial agents found in Japan (E. muris and a recently discovered Ehrlichia species detected from Ixodes ovatus) in blood samples from dogs in Yamaguchi and Okinawa Prefecture, Japan. Partial sequence of E. platys was detected from 1 of 67 dogs (1.5%) tested from Yamaguchi Prefecture and 24 out of 87 (27.6%) in the subtropical Okinawa Prefecture. Dogs in Okinawa and Miyako Islands had a higher positive rate (69.2 and 45.0%, respectively) than Ishigaki Island (11.1%). Another dog in Yamaguchi Prefecture had a positive PCR reaction to the Ehrlichia sp. detected from I. ovatus. No other Ehrlichia were found in these samples.     

Response of Nkedi Zebu and Ankole cattle to tick infestation and natural tick-borne,helminth and trypanosome infections in Uganda

A cross-sectional study was conducted in Soroti district of Uganda to establish important traits of Nkedi Zebu and Ankole cattle regarding their production performance responses to natural infections of trypanosomes, gastrointestinal nematodes, Theileria parva, Babesia bigemina, Anaplasma marginale and tick infestations. Over four visits between October 2006 to August 2007, tick counts were performed and blood, faecal samples and sera were collected from the Nkedi Zebu (295) and Ankole (165) cattle from 86 herds in six locations per visit. Low parasitological prevalence of trypanosome infection (<6%) and high prevalence of gastrointestinal nematode infections (>30%) with low faecal egg counts (110–300 eggs per gramme (EPG)) were observed in the Nkedi Zebu and Ankole cattle. Both breeds had high, moderate and low mean counts of Rhipicephalus appendiculatus (18.0–24.0), Rhipecephalus (Boophilus) decoloratus (3.6–10.3) and Amblyomma variegatum ticks (1.7–4.3), respectively. In addition, both breeds had similar mean packed cell volumes (26.4–31.2) and a similar percentage of animals were anaemic (14.5–36.6%). The Nkedi Zebu cattle further had higher mean optical density (OD) values for antibodies against T. parva (1.093–1.445) and A. marginale infections (0.573–0.583), and significantly (P < 0.001) higher mean OD values of antibodies against B. bigemina infections (1.07–2.175) than the Ankole cattle: T. parva (1.030–1.302); A. marginale (0.442–0.603) and B. bigemina infections (0.863–2.154). The Ankole cows produced significantly more (P < 0.001) milk per day (2.68 L) than the Nkedi Zebu cows (1.98 L), and the Ankole oxen had significantly higher (P < 0.05) draught power output (2.57 days/acre) than the Nkedi Zebu oxen (2.93 days/acre). Liveweights of calves aged 0–12 months of both breeds were comparable, suggesting that the Nkedi Zebu and Ankole cattle under similar disease challenge exhibited similar growth rates. In conclusion, the Nkedi Zebu cattle seem to possess a higher degree of disease resistance against endemic parasitic diseases, while the Ankole cattle seem to possess a moderate degree of disease resistance coupled with a moderate production potential.     

Cryptosporidium infection in dogs in Osaka,Japan

Cryptosporidium parvum is a zoonotic pathogen composed of genetically distinct but morphologically identical genotypes. Recent molecular study indicates that dogs may transmit the cattle genotype, which is known to be pathogenic to humans. Although large-scale studies of Cryptosporidium infection in dogs have been performed in several countries, the isolates were not accurately identified because of the lack of a method for molecular analysis. It is important to identify the isolates harbored in dogs, which come in close contact with humans, in order to control human cryptosporidiosis. The aim of the present study was to calculate the prevalence of Cryptosporidium infection in dogs in Osaka city, Japan, and to characterize the isolates molecularly. The prevalence was determined to be 9.3% (13/140) by PCR. All isolates were found to be Cryptosporidium canis (previously known as the dog genotype), which is thought to be non-pathogenic in humans, based on the sequencing of diagnostic fragments. These results indicate that PCR-based diagnostic methods are a useful tool for the diagnosis and molecular epidemiology of Cryptosporidium infection in dogs, and that dogs living in Osaka are not a significant reservoir for human cryptosporidiosis. It is unclear why C. canis is dominant in dogs. Further study is required to understand this partial parasitism.     

Epizootiological study of tick-borne encephalitis virus infection in Japan

Tick-borne encephalitis virus (TBEV) is a zoonotic agent causing severe encephalitis in humans. Rodent species that are potential hosts for TBEV are widely distributed in various regions in Japan. In this study, we carried out large-scale epizootiological surveys in rodents from various areas of Japan. A total of 931 rodent and insectivore sera were collected from field surveys. Rodents seropositive for TBEV were found in Shimane Prefecture in Honshu and in several areas of Hokkaido Prefecture. These results emphasize the need for further epizootiological and epidemiological research of TBEV and preventive measures for emerging tick-borne encephalitis in Japan.     

Survey of tick infestation in small ruminants of Miesso district,West Harergie,Oromia Region,Ethiopia

A survey was conducted to identify tick species and determine the prevalence of tick infestation in small ruminants of Miesso District, West Harergie Zone. Collection and identification of the ticks were undertaken from November 2007 to April 2008. All visible individual adult ticks were collected from the body of 328 goats and 40 sheep. The prevalence of tick infestation in goats and sheep was found to be 89.9% and 87.5%, respectively. In this study, ten species of ticks which grouped under four genera were identified. The most abundant species found in this study were Boophilus decoloratus (60%), Rhipicephalus pulchellus (25.1%), and Amblyomma gemma (11%). Hyalomma dromedarii was the minor species observed on goats. The difference in the prevalence of tick infestation between sheep and goats was not statistically significant (Χ ²   = 0.22, p = 0.63) but found to be statistically significant between male and females (Χ ²  = 9.8, p = 0.003). Attention should be given to the control and prevention of ticks, since they cause sever damage to the skins of small ruminants and thereby reduce the foreign exchange of the country; they also transmit some diseases which can cause sever loss to the productivity of these animals.     

Survey of ixodid tick species on domestic cats in Japan

Various species of ixodid ticks, attached to domestic cats in Japan, were identified in spring (April to June) and autumn (September to November). In the spring, a total of 282 ticks, including 61 larvae, 70 nymphs, 127 females and 24 males were collected from 126 cats. Of these, 264 were identified up to the species level. In the spring, Haemaphysalis longicornis was the most frequently (39.7%, 50/126) found tick species on feline hosts, followed by Ixodes ovatus (35.0%, 44/126), Ixodes nipponensis (15.9%, 20/126) and Haemaphysalis flava (9.5%, 12/126). Small numbers of Haemaphysalis megaspinosa, Haemaphysalis japonica, Ixodes persulcatus, Ixodes granulatus and Amblyomma testudinarium were also recovered. H. longicornis was the most frequently found tick species on cats around riversides or river basins, while I. ovatus and I. nipponensis were more frequently found on cats kept near woodland or related areas. I. nipponensis was more frequently found on castrated males. No major statistical differences in the frequency of tick attachment among sex, age or hair length for the three major tick species were found. Of 205 ticks including 173 (84.4%) larvae, 27 (13.2%) nymphs, 4 (2.0%) females and 1 (0.5%) male recovered from 62 cats in autumn, only 32 (15.6%) were identified. Most of the larvae were fully- or partly-engorged Haemaphysalis spp., and it was difficult to identify them further by morphological characterization.     

Experimental transmission of granulocytic ehrlichial organisms in dogs

Canine granulocytic ehrlichial organisms were transmitted from an infected dog from Missouri to two male, 10-month-old dogs by an intravenous injection of whole blood. Physical or behavioral abnormalities were not detected during the 98 days of evaluation other than a mild pyrexia from Day 18 to 20. Ehrlichial morulae were found in blood granulocytes of Dog 1 from Day 13 to 44 and of Dog 2 from Day 14 to 34 with the peak rickettsemia occurring on Day 16 for both dogs. By Day 21 after inoculaiuon, both dogs had positive titers to Ehrlichia canis. The highest titers for both dogs were found 63 days after inoculation, after which the titers decreased. Most of the hematologic abnormalities (i.e., neutropenia, lymphocytosis, thrombocytopenia) and fever occurred between 18 and 24 days after inoculation. The pathologic bases of these abnormalities were not investigated but their concurrent presence suggested an association with the dogs' immunologic responses to the granulocytic ehrlichial agent. Results from the study indicated that the canine granulocytic ehrlichial agent of Missouri may produce subclinical infections and suggested that dogs may be able to clear the organism without antimicrobial therapy.     

Cutaneous lesions in dogs with Dirofilaria (Nochtiella) repens infestation and concurrent tick-borne transmitted diseases

A pruritic dermatitis characterized by the presence of erythema, papules, focal or multifocal alopecia, crusting and nodules was seen in 28 dogs with Dirofilaria repens microfilariae infestation in an endemic area in north-west Italy. Previous unsuccessful nonspecific antipruritic treatments, flea control and restricted diet were recorded in 53.6% of the patients. Both the Knott and the antigen tests were negative for Dirofilaria immitis and Acanthocheilonema reconditum. Concurrent babesiosis and/or canine granulocytic ehrlichiosis was also diagnosed in many affected dogs. Preliminary treatment of the concurrent diseases was followed by specific filaricide treatment. The cutaneous lesions, although slightly improved with the initial treatment, resolved completely with macro- and microfilaricide treatment. Although D. repens may be an opportunistic pathogen, this parasite should not be considered as harmless as previously thought and its potential pathogenic role in causing cutaneous lesions in dogs should be considered. Furthermore, it has a zoonotic importance as human cases have been reported worldwide.     

Molecular survey of Babesia infection in dogs in Okinawa, Japan

A total of 80 free-roaming dogs on Okinawa Island, Japan, were examined for Babesia infection using the polymerase chain reaction (PCR) and sequence analysis. Of 80 samples, 12 were positive in a Babesia genus-specific PCR. Consequent species-specific PCR for B. canis and B. gibsoni revealed that 5 (6.3%) and 7 (8.8%) dogs were infected with B. canis and B. gibsoni, respectively. Sequence analysis of the PCR products revealed that the 18S rRNA gene sequence of B. canis detected from dogs in Okinawa was very close to B. canis vogeli with sequence similarity of 99.94%.     

Survey of flea infestation in dogs and cats in the United Kingdom during 2005

During 2005, 31 uk veterinary practices participated in a survey of flea infestation, during which 2653 dogs and 1508 cats were examined for evidence of flea infestation and skin disease compatible with flea allergy dermatitis (fad). The prevalence of flea infestation in the cats was 21.09 per cent, significantly (P<0.001) higher than in the dogs (6.82 per cent). The prevalence of skin lesions compatible with fad in the cats (8.02 per cent) was also significantly (P<0.001) higher than in the dogs (3.32 per cent). Flea infestations were more common in households with cats and with more than one pet. Of 467 fleas identified from the cats, 462 (98.93 per cent) were Ctenocephalides felis, one was Ctenocephalides canis, one was Archaeopsylla erinacei, two were Pulex irritans, and one was Spilopsyllus cuniculi. Of 336 fleas identified from the dogs, 313 (93.15 per cent) were C felis, five were C Canis, 12 were A erinacei, five were P irritans, and one was Ceratophyllus (Nosophyllus) fasciatus. Almost half of the owners of the dogs and cats were unaware of their pet's flea infestation. The overall prevalence of fleas and/or skin lesions that could potentially be compatible with fad was 7.46 per cent in the dogs and 22.28 per cent in the cats.     

Effects of tick infestation and tick-borne disease infections (heartwater,anaplasmosis and babesiosis) on the lactation and weight gain of mashona cattle in South-Eastern Zimbabwe

Summary The effects of ticks and tick-borne disease infections on the lactation and weight gain of Mashona cattle were studied at Mbizi Quarantine Station in the south-eastern lowveld of Zimbabwe. Twenty-nine Mashona cows were allocated to 2 balanced groups and kept in separate paddocks at a stocking rate of one animal per 8 ha. One group received regular acaricide treatment to control bont(Amblyomma hebraeum) and other ticks. The other group was left untreated. The cows were artificially inseminated. The acaricide-treated cows and calves were essentially tick free throughout the experiment, while the untreated cows and calves were continuously tick infested. There was a drought-related decline in tick infestations in the second year of the experiment. Antibodies toCowdria ruminantium, Babesia bigemina andAnaplasma marginale were detected in cows and calves from both groups, though the untreated group had significantly higher titres toC. ruminantium (P < 0·001). The total, measured amount of milk suckled by untreated calves was significantly more than treated calves (273 kg vs. 241 kg,P 0·05). By interpolating between the twice weekly measurements, it was calculated that over the entire lactation untreated calves suckled an average of 935 kg/hd vs. 837 kg/hd for the treated group. There were no statistical differences in the weights of the 2 groups of calves at birth, weaning, 180 and 210 dayspost partum (P < 0·05). For cows, there were no statistically significant differences in gestation periods (288 vs. 279 days), reconception rates or weight patterns over time (P < 0·05). The results show that intensive acaricide treatment in areas of Zimbabwe where heartwater is enzootically stable is uneconomical. The maintenance of enzootic stability for tick-borne diseases through minimal tick control is clearly a more economic and practical control option.

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Efecto De La Infestacion Por Garrapatas Y De Las Enfermedades Transmitidas Por Garrapatas (Cowdriasis, Anaplasmosis Y Babesiosis) Sobre La Produccion De Leche Y El Crecimiento Del Ganado Vacuno De Raza Mashona En El Sureste De Zimbabwe

Resumen Se estudiaron los efectos de la infestación por garrapatas y de las enfermedades transmitidas por garrapatas sobre la producción de leche y el crecimiento del ganado vacuno de raza Mashona. El estudio se realizó en la Mbizi Quarantine Station, situada en la zona de estepa baja del sureste de Zimbabwe. Un total de 29 vacas Mashona se distribuyeron en 2 grupos y se mantuvieron en 2 parcelas separadas a una densidad de 8 ha por animal. Uno de los grupos fue tratado regularmente con acaricidas para controlar la infestación porAmblyomma hebraeum y otras garrapatas. El otro grupo no recibió ningún tratamiento. Las vacas se inseminaron artificialmente. Las vacas y terneros del grupo tratado con acaricida estuvieron libres de garrapatas durante todo el experimento, mientras que los animales del grupo que no fue tratado estuvieron continuamente infestados. Durante el segundo año del experimento se produjo una disminución en el nivel de infestacion debido a la sequia. Tanto en los terneros como en las vacas de ambos grupos se detectaron anticuerpos frente aCowdria ruminantum, Babesia bigemina y Anaplasma marginale, si bien los animales del grupo no tratado tuvieron titulos de anticuerpos frente aC. ruminantum màs altos que los animales tratados (P < 0·001). La cantidad total de leche media consumida por los terneros fue significativamente mayor en el grupo no tratado que en el grupo tratado (273 kgs vs. 241 kgs,P < 0·05). Mediante interpolación a partir de las dos mediciones semanales se calculó que a lo largo de toda la lactación los terneros del grupo no tratado consumieron una media de 935 kgs de leche, mientras que los terneros del grupo tratado consumieron una media de 837 kgs. No hubo diferencias significativas entre los dos grupos en los pesos de los terneros al nacimiento, al destete, a los 180 días postparto ni a los 210 días postparto (P < 0·05). No hubo diferencias significativas entre los dos grupos de vacas en la duración de la gestación (288 vs 279 dias), tasa de fertilidad ni variaciones de peso a lo largo del tiempo (P < 0·05). Los resultados indican que el tratamiento intensivo con acaricidas en zonas de Zimbabwe donde la cowdriasis es enzoóticamente estable no resulta económico. Mantener la estabilidad enzoótica de las enfermedades transmitidas por garrapatas mediante un control mínimo de las garrapatas es una alternativa claramente màs pràctica y económica.

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Effet De L'infestation Ixodidienne Et Des Maladies Transmises Par Tiques (Cowdriose, Anaplasmose Et Babesiose) Sur La Lactation Et Le Gain De Poids Chez Le Betail Mashona Du Sud-Est Du Zimbabwe

Résumé On a étudié à la station de quarantaine de Mbizi, dans le bas veld du sud-est du Zimbabwe, les effets des infestations ixodidiennes et des maladies à tiques sur la lactation et le gain de poids chez le bétail Mashona. Vingt-neuf vaches Mashona ont été réparties en 2 groupes équilibrés et placés dans des enclos séparés au taux d'occupation d'un animal pour 8 ha. Un groupe a reçu un traitement acaricide normal pour contrôlerAmblyomma hebraeum et les autres tiques; l'autre groupe a été laissé sans traitement. Les vaches traitées par acaricide, ainsi que leur veaux, sont restées pratiquement libres de tiques pendant toute l'expérience alors que les vaches et veaux non traités étaient infestés en permanence. Pendant la seconde année de l'expérience s'est produit une baisse de l'infestation ixodidienne concommitante d'une sécheresse. Des anticorps contreCowdria ruminantium, Babesia bigemina etAnaplasma marginale ont été detectés chez les vaches et les veaux des deux groupes bien que le groupe non traité ait des titres antiC. ruminantium significativement plus élevés (P < 0,001). La quantité total mesurée de lait ingéré par les veaux non traités a été significativement plus elevée que celle des veaux traités (273 kg contre 241 kg,P < 0,05). Par interpolation entre deux mesures bi-hebdomadaires, on a calculé que, pour toute la période de lactation, les veaux non traités ont tété une moyenne de 935 kg/tête contre 837 kg/tête pour le groupe traité. Aucune différence statistiquement significative n'a été noté pour les deux groupes de veaux, à la naissance, au sevrage, à 180 et à 210 jours aprés la naissance (P < 0,05). Chez les vaches, on n'a noté aucune différence statistiquement significative dans les périodes de gestation (288 et 279 jours), les taux de reconception ou les courbes de poids (P < 0,05). Ces résultats montrent que, dans les régions du Zimbabwe où la cowdriose est en stabilité enzootique, les traitements acaricides intensifs sont antiéconomiques. Le maintien de la stabilité enzootique pour les maldies transmises par tiques gráce a un contrôle anti-ixodidien minimum est, à l'évidence, l'option de contrôle la plus économique et la plus pratique.

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Deceased     

Incidence of Salmonella infection in healthy dogs in Gifu Prefecture,Japan

A total of 1,013 feces samples and 8 mesenteric lymphonodus samples obtained from apparently healthy dogs were examined for the incidence of salmonella infection. One strain of S. typhimurium (ST) was isolated from feces of one dog, and S. enteritidis (SE) was isolated from the mesenteric lymphonodus of one dog. Sera obtained from 330 apparently healthy dogs were examined for Salmonella antibodies using an ELISA with heated whole cells of SE and ST. Fifty-one of the 330 serum samples were considered to be positive for salmonella antibodies, including 12 which were SE-positive and 39 which were ST-positive. These results indicate that dogs cause possible environmental problems as Salmonella carriers.     

Natural Anaplasma marginale infection in 2 cattle herds with different levels of Boophilus microplus tick infestation

The dynamics of natural infections by Anaplasma marginale was studied in two adjacent dairy farms with different levels of Boophilus microplus infestation. The farms were located in the enzootic area of bovine anaplasmosis of the Northwest of Argentina. The study was carried out in 35 calves from birth in March-August 1985 to March 1986. The infection rate by A. marginale was evaluated by the observation of blood films and by determination of specific antibodies. The degree of infestation by B. microplus was also evaluated. The tick was found all over the year in farm A with peaks of 100 and 95% of infested calves in October and January, respectively. In farm B, B. microplus was found only in December and January with a maximum of 50% of infested calves. Natural infections by A. marginale started in June until the end of the study when 89% (farm A) and 81% (farm B) of the calves proved to be infected. According to the active serological reactors, the rate rose to a maximum of 85% (farm A) and 81% (farm B) at the end of the study. It is remarkable that 69% of primo-infections by A. marginale in farm B occurred when B. microplus was absent. Moreover, no direct relationship between the peaks of tick infestation and primo-infections with the rickettsie was detected in farm A. The authors concluded that B. microplus could have less importance in the transmission of A. marginale than previously assumed under the local conditions.     

Survey and molecular characterization of Cryptosporidium and Giardia spp. in owned companion animal, dogs and cats, in Japan

Compared with other countries, surveys of these parasites have been rarely performed in companion animals of Japan in spite of their significance for public health. Here, we investigated pet dogs and cats in Japan for the first time, and genetically analyzed the isolates to evaluate the risk of zoonotic infections. Seventy-seven fecal samples were collected from privately owned dogs and 55 samples from owned cats in Osaka city, Japan. Cryptosporidium oocysts were identified in 3/77 dogs (3.9%) and 7/55 cats (12.7%), and Giardia infection in 2/77 dogs (2.6%) and 1/55 cats (1.8%). Amplification of the target regions for genotyping was successful, Cryptosporidium isolates in dogs and cats were identified as C. canis and C. felis, respectively, and those of Giardia in dogs and cats were G. intestinalis Assemblages D and F. The discharge period of the oocysts varied within 3-16 weeks and that of the cysts was 12 weeks. To date, zoonotic types of both parasites have been identified in other animals in Japan, and further large-scale studies are needed to determine the distribution of zoonotic genotypes in these animals, especially those closely associated with humans.     

Incidence of canine Babesia gibsoni infection and subclinical infection among Tosa dogs in Aomori Prefecture, Japan

To identify the incidence of Babesia gibsoni (B. gibsoni) in Aomori Prefecture, northeastern Japan, dogs with acute B. gibsoni infection were investigated at the Animal Teaching Hospital, Kitasato University, between April 2002 and March 2003. Eighteen dogs with acute B. gibsoni infection were recognized; they were all male dogs of the fighting dog breed Tosa. Their platelet counts were below normal and their packed cell volumes (PCVs) were at various levels. We collected blood samples from 141 Tosa dogs from Aomori Prefecture and used polymerase chain reaction assay to investigate the incidence of subclinical B. gibsoni infection. We also looked into the serological abnormalities associated with thrombocytopenia or anemia in subclinical infection. Forty-one of 87 dogs (47.1%) with histories of dog fighting, and one dog of 54 without a history of dog fighting were positive for B. gibsoni; that is, 42 of 141 dogs (29.8%) showed a positive result. The mean platelet counts of dogs with subclinical infection were significantly lower and levels of anti-platelet IgG were significantly higher than levels for dogs without infection. Anti-erythrocyte membrane IgG levels were significantly higher in dogs with subclinical infections, although mean PCVs were not significantly different. Tosa dogs from Aomori Prefecture, Japan, were highly infected with B. gibsoni subclinically and this pathogen might be successfully transmitted during dog fighting. Dogs with subclinical infections were at risk of chronic thrombocytopenia, which may be due to autoimmune mechanisms.     

Serosurvey of Ehrlichia canis and Hepatozoon canis infection in dogs in Yamaguchi Prefecture, Japan

Antibodies to Ehrlichia canis and Hepatozoon canis in dogs at the Animal Hospital in Yamaguchi University were surveyed and potential risk factors for both pathogens were evaluated. Among 430 dogs examined, 20 (4.7%) and 18 (4.2%) dogs showed positive findings for E. canis and H. canis, respectively. Neither, sex nor age was associated with the seropositivity of either pathogen, but the positive rate in dogs kept outside was slightly higher than that in dogs kept inside for both pathogens. A higher seropositive reaction to E. canis and H. canis was observed in dogs that lived in certain cities and towns. Beagles, golden retrievers and pointers had higher seropositivity than other breeds in E. canis, whereas shibas, akitas, beagles, pointers and mongrels had higher positive rates than other breeds in H. canis.     

Phylogenetic identification of Cystoisospora spp. from dogs, cats, and raccoon dogs in Japan

Cystoisospora spp. from feces in dogs, cats, and raccoon dogs were isolated, sequenced at the small subunit ribosomal RNA gene locus and compared to other Cystoisospora spp. Cystoisospora oocysts from dogs and raccoon dogs were morphologically similar with those of C. ohioensis, and cat isolates were similar with those of C. felis. The sequences from dogs and raccoon dogs, and cats have a homology with C. ohioensis and C. felis, respectively. Phylogenetic analysis of the DNA sequences showed that the dog and raccoon dog isolates were nested in a clade with other Cystoisospora spp. including C. ohioensis, C. belli, and C. orlovi. The cat isolate formed a sister group with C. felis that was a separate clade from the dog and raccoon dog group. We report sequence variation in these Cystoisospora sequences and have identified raccoon dogs as another carnivore host for Cystoisospora spp. infecting dogs.