Prevalence of antibodies against Toxoplasma gondii and Neospora caninum in Hungarian red foxes (Vulpes vulpes)

In the present study we have investigated the seroprevalence to the protozoan parasites Toxoplasma gondii and Neospora caninum in 337 red foxes (Vulpes vulpes) from 16 out of 19 counties in Hungary. The foxes were originally collected within a National vaccination program against rabies. Antibodies to T. gondii were detected in as many as 228 (68%) of the foxes using a commercial direct agglutination test (DAT). In an indirect iscom ELISA, five foxes (1.5%) were positive for antibodies against N. caninum. The high prevalence of foxes positive for T. gondii might be explained by the widespread occurrence of the parasite in the diet of foxes. As a contrast, latent infections of N. caninum among red foxes in Hungary are much less common.     

Seroprevalences of antibodies to Neospora caninum and Toxoplasma gondii in zoo animals

Neospora caninum is an apicomplexan parasite that causes neuromuscular disease in dogs and abortions in cattle. Little is known about the prevalence of antibodies to this parasite in zoo animals. Sera from 556 animals, from 13 Czech and Slovak zoos were tested for antibodies to N. caninum and Toxoplasma gondii by indirect fluorescent antibody test. Antibodies to N. caninum were found in 31 of 556 zoo animals (5.6%), representing 18 of 114 species tested: Eurasian wolf (Canis lupus lupus), Maned wolf (Chrysocyon brachyurus), fennec (Vulpes zerda), cheetah (Acinonyx jubatus), jaguarundi (Herpailurus yaguarondi), Eurasian lynx (Lynx lynx), Indian lion (Panthera leo goojratensis), fisher (Martes pennanti), blackbuck (Antilope cervicapra), European bison (Bison bonasus), lechwe (Kobus leche), African buffalo (Syncerus caffer caffer), eland (Taurotragus oryx), sitatunga (Tragelaphus spekei gratus), Thorold's deer (Cervus albirostris), Eastern elk (C. elaphus canadensis), Vietnam sika deer (C. nippon pseudaxis) and Père David's deer (Elaphurus davidianus). Titres ranged from 1:40 to 1:2560. The highest prevalence 50% was found in family mustelidae of the order carnivora. Antibodies to T. gondii were detected in 193 of 556 zoo animals (34.7%) representing 72 of 114 species tested, with titres ranging from 1:40 to 1:40960. The highest prevalence 100% was found in families: hyaenidae, mustelidae, ursidae and viveridae of the order carnivora. The results of this study indicate that zoo animals have more exposure to T. gondii than to N. caninum. It is the first report of seroprevalence of antibodies to N. caninum in European zoo animals.     

Red foxes (Vulpes vulpes) are a natural intermediate host of Neospora caninum

The present study was undertaken to determine if red foxes are natural intermediate and/or definitive host for Neospora caninum and to study the importance of infection of N. caninum in this species in North-eastern Spain. Faecal samples and brain tissues were obtained from 122 foxes from 21 rural areas of Catalonia. Faeces collected were examined for parasite eggs and coccidian oocysts using sucrose flotation. For PCR-based diagnosis of N. caninum in brain tissues, the specific genomic Nc5 region was selected as the target sequence for DNA amplification. To control for PCR failure and facilitate identification of truly negative samples, the competitor pNc5C molecule was added to all negative samples in a second round of PCR reactions. Of the 122 foxes analysed, 13 (10.7%) were positive by PCR for N. caninum. Signal intensities of all positive samples were relatively weak with the exception of one sample from a 3-month male animal, that also showed the highest repeatability. No differences were observed by sex, age or area of sampling analysis. Detection of stages of N. caninum in brain from naturally infected red foxes demonstrated that red foxes are a natural intermediate host for N. caninum. Faecal samples were analysed for the presence of N. caninum oocysts, however, no oocysts compatible with N. caninum were found. A widespread latent infection of red foxes in North-eastern Spain found in the present study indicates that red foxes could have a very important role in the epidemiology of neosporosis in our area.     

Seroprevalences of Toxoplasma gondii and Neospora sp. infections in Swedish horses

Sera from 414 Swedish horses were investigated for the presence of antibodies to Toxoplasma gondii and Neospora sp. by the T. gondii direct agglutination test (DAT), and an Neospora caninum iscom-ELISA. Five sera (1%) had a titre >1:40 in DAT, but when analysed by immunoblotting against T. gondii antigens only two of them were positive, giving a seroprevalence of 0.5%. Since the Neospora iscom ELISA had not been validated for equine sera it was used for an initial screening, and all sera with an optical density exceeding 0.200 absorbance units were selected for further investigation by immunoblot analysis. Of the 39 sera tested by immunoblotting, four reacted with at least two of the immunodominant Neospora antigens recognized by the positive control sera and were judged as positive, resulting in a seroprevalence of 1%. This is the first evidence of Neospora infection in Swedish horses. The study illustrates the necessity of critically evaluating results of serological analyses performed by methods that are not validated for the animal species under investigation.     

Study on the prevalence of Toxoplasma gondii and Neospora caninum and molecular evidence of Encephalitozoon cuniculi and Encephalitozoon (Septata) intestinalis infections in red foxes (Vulpes vulpes) in rural Ireland

Thoracic fluid (pleural fluid and clotted blood) from 206 foxes were examined for antibodies to Toxoplasma gondii and 220 thoracic fluid samples were tested for Neospora caninum antibodies using indirect immunofluorescent antibody tests (IFAT). A total of 115 (56%) and six (3%) foxes had antibodies to T. gondii and N. caninum, respectively. The brains from 148 foxes were examined for histological lesions and pathological changes suggestive of parasitic encephalitis were observed in 33 (22%). Two thirds of these foxes had antibodies to T. gondii and one fox had antibodies to both T. gondii and N. caninum. PCR assays carried out on DNA extracted from the 33 brains with histological lesions were negative for N. caninum but one of the brains was positive for T. gondii. Microsporidian DNA was also amplified from the brains of two of these foxes. Sequencing these amplicons revealed 100% homology with Encephalitozoon (Septata) intestinalis in one fox and Encephalitozoon cuniculi in the second fox. This is the first report of Encephalitozoon infections in wildlife in Ireland.     

Seroprevalences of Neospora caninum and Toxoplasma gondii in nondomestic felids from southern Africa.

Sera from 68 nondomestic captive and free-ranging felids from southern Africa were tested for antibodies to Neospora caninum and Toxoplasma gondii by the indirect fluorescent antibody test. Four of the 68 (5.9%) serum samples were positive for antibodies to N. caninum, with titers ranging from 1:50 to 1:200. All other animals were negative for antibodies to N. caninum at a dilution of 1:50. Fifty of the 68 (74%) serum samples tested positive for antibodies to T. gondii, with titers ranging from 1:50 to 1:26,500. Four animals tested positive for antibodies to both N. caninum and T. gondii. None of these animals displayed clinical signs of disease. Results of this study indicate that nondomestic felids in southern Africa have been exposed to, and are likely infected with, N. caninum and T. gondii.     

Prevalence of Antibodies to Neospora caninum and Toxoplasma gondii in Swedish Dogs

Neospora caninum is a newly described coccidian parasite which has been found in various species such as the dog, cattle, horse, sheep and goat. Morphologically it resembles Toxoplasma gondii with which it is related (Holmdahl et al. 1994), and with which it has earlier been confused. The life cycle of N caninum is only partially known. Tachyzoites and tissue cysts are the only known stages of the parasite, and transplacental transmission is the only known route of infection. Subclini-cally infected dams can transmit the parasite to their fetuses and successive offspring from the same mother might be born infected (Dubey et al. 1990b). Clinical neosporosis is mostly seen in pups or young dogs, and the majority or all pups in a litter are often affected. The disease is characterized by ascending paralysis of the legs, with the hind legs more severely affected than the front legs, paralysis of the jaw, difficulty in swallowing and muscle flaccidity and atrophy (Dubey 1992, Dubey & Lindsay 1993). Fatal infections with N caninum in dogs have been reported from many countries, e.g. Norway (Bjerkäs & Presthus 1988), USA (Dubey et al. 1988), Sweden (Uggla et al. 1989a,b) and the United Kingdom (Dubey et al. 1990a). Serological surveys for antibodies to N. caninum in dogs from Kansas, USA and England have shown a prevalence of 2 and 13%, respectively (Lindsay et al. 1990, Trees et al. 1993).     

Helminths of red foxes (Vulpes vulpes) in Denmark

An epidemiological study of helminths in 1040 red foxes collected from various localities in Denmark during 1997-2002, revealed 21 helminth species at autopsy, including nine nematode species: Capillaria plica (prevalence 80.5%), Capillaria aerophila (74.1%), Crenosoma vulpis (17.4%), Angiostrongylus vasorum (48.6% from Northern Zealand (endemic area)), Toxocara canis (59.4%), Toxascaris leonina (0.6%), Uncinaria stenocephala (68.6%), Ancylostoma caninum (0.6%), and Trichuris vulpis (0.5%); seven cestodes: Mesocestoides sp. (35.6%), a number of Taeniid species (Taenia pisiformis, T. hydatigena, T. taeniaeformis, T. crassiceps, and unidentified Taenia spp.) (22.8%), and Echinococcus multilocularis (0.3%); four trematodes: Alaria alata (15.4%), Cryptocotyle lingua (23.8%), Pseudamphystomum truncatum (3.6% from Northern Zealand), and Echinochasmus perfoliatus (2.4% from Northern Zealand); one acanthocephalan: Polymorphus sp. (1.2%). Significant difference in prevalence was found for T. canis and A. vasorum according to host sex, and for T. canis, U. stenocephala, Mesocestoides sp., Taenia spp., A. alata, A. vasorum, and Capillaria spp. according to age groups (adult, young or cub). Prevalence and average worm intensity for each helminth species varied considerably according to geographical locality, season, and year. Aggregated distribution was found for several helminth species. The two species E. multilocularis and E. perfoliatus are first records for Denmark.     

Prevalence of antibodies against Neospora caninum and Toxoplasma gondii in dogs and foxes in Austria

Sera from 1770 dogs and 94 red foxes from Austria were examined for antibodies against Neospora caninum using the indirect immunofluorescent antibody test (IFAT). 3.6% of the dogs were seropositive with titres ranging from 1:50 to 1:6400. Dogs from rural areas were significantly more often seropositive for N. caninum than those from the urban area of Vienna (5.3% versus 2.1%). There were no significant differences in sex or breed, but a slight increase in seropositivity with age was apparent, indicating postnatal infection. None of the foxes had antibodies against N. caninum. Additionally, sera from 242 dogs and 94 foxes were examined for antibodies against Toxoplasma gondii using the IFAT. Thirty-five percent foxes and 26% of the dogs were positive; 1.7% of the dogs were positive for both parasites. This is the first report of the prevalence of N. caninum infections in dogs and foxes in Austria.     

Ectoparasite infestations of red foxes (Vulpes vulpes) in Hungary

A survey was carried out to investigate the ectoparasite infestations of 100 red foxes (Vulpes vulpes) in Hungary. The overall prevalence of flea and tick infestation was high (62 and 86% with the dominance of the anthropophil Pulex irritans and Ixodes ricinus), but the number of parasites was low to moderate. Felicola vulpis was not found in the present study, and the prevalence of Otodectes cynotis was only 2%. Based on prevalence (21%), mange lesion scores, and the negative correlation between lesion scores and condition of foxes, Sarcoptes infestation should be considered as the most important parasitosis of foxes in Hungary. Besides the ecological significance of these parasites, the high overall prevalence of mange and anthropophil flea and tick infestations of foxes and the appearance of these animals in the synanthropic environment as a result of the increasing population size, may result in the increasing incidence of flea, tick and accidental mite infestation of man and domestic animals, and may enhance the transmission rate of some vector-borne diseases.     

Extraintestinal nematode infections of red foxes (Vulpes vulpes) in Hungary

A survey was carried out to investigate the prevalence and worm burden of extraintestinal nematodes in 100 red foxes (Vulpes vulpes) of Hungary. The overall prevalence of nematode infections of the respiratory tract was 76%. Eucoleus aerophilus (Capillaria aerophila) was the predominant species (66%), followed by Crenosoma vulpis (24%), Eucoleus (Capillaria) b?hmi (8%) and Angiostrongylus vasorum (5%). Pearsonema (Capillaria) plica was found in 52% of the urinary bladders. In 3% of the foxes, Trichinella britovi was present in muscle samples. The high prevalence of lungworms and P. plica and the fox colonisation in urban areas may enhance the prevalence of these nematode infections in domestic dogs and cats, and the flow of T. britovi from the sylvatic cycle to the domestic cycle, enhancing the risk of infections in humans.     

High seroprevalence of Neospora caninum in the red fox (Vulpes vulpes) in the Pyrenees (NE Spain)

Antibodies to Neospora caninum were determined in the red fox (Vulpes vulpes) in the Pyrenees, north-eastern Spain. Sera from 53 red foxes (29 male and 24 female) were tested using a Neospora agglutination test (NAT). Seroprevalence at dilutions of 1/40, 1/80 and 1/160 was 69.8%, 47.2% and 7.5%, respectively. Sex differences were significant only at a dilution of 1:40. The high seroprevalence observed in red fox suggests that this species is highly exposed to N. caninum in this area.     

Prevalence of antibodies to Neospora caninum and Toxoplasma gondii in gray foxes (Urocyon cinereoargenteus) from South Carolina

Little is known about the epidemiology of Neospora caninum in wild mammal populations. It has been suggested that a sylvatic cycle exists for N. caninum. Dogs and potentially other canids are a definitive host for N. caninum. The present study was done to determine the prevalence of antibodies to N. caninum in a population of gray foxes (Urocyon cinereoargenteus) from a nonagricultural setting in South Carolina. We also determined the prevalence of antibodies to Toxoplasma gondii in these animals. Antibody levels were measured in direct agglutination tests using either N. caninum or T. gondii formalin-fixed tachyzoites as antigen. Four (15.4%) of the 26 gray foxes had titers to N. caninum. Titers to N. caninum were low being 1:25 in three gray foxes and 1:50 in the fourth gray fox. Antibodies to T. gondii were observed in 16 (61.5%) gray foxes. Titers to T. gondii were usually >1:50 and two gray foxes had titers of 1:1600. Results of this study indicate that gray foxes have more exposure to T. gondii than to N. caninum in this environment.     

Occurrence of Giardia and Cryptosporidium in Norwegian red foxes (Vulpes vulpes)

Faecal samples from 269 Norwegian wild red foxes (Vulpes vulpes) shot during the hunting season (October-April) in 2002-2004 were examined for the presence of Giardia and Cryptosporidium. Cryptosporidium oocysts were detected in samples from 6 (2.2%) of the foxes, and Giardia cysts in 13 (4.8%) of the foxes. The prevalence of Giardia infection was significantly higher in juvenile male foxes than in adult male foxes, but no other significant differences between age and sex were found. No significant differences in prevalence related to geographical origin of animals were found. Insufficient nucleated Cryptosporidium oocysts were isolated for successful PCR, but genotyping of Giardia duodenalis isolates from seven foxes demonstrated a high degree of heterogeneity amongst them, with all isolates belonging to the zoonotic Assemblages A and B.     

Distribution of Crenosoma vulpis and Eucoleus aerophilus in the lung of free-ranging red foxes (Vulpes vulpes).

Crenosoma vulpis and Eucoleus aerophilus are nematode parasites that can cause verminous pneumonia in wild carnivores. There is a paucity of information regarding the distribution of parasites in the lungs and the relationship between histopathological and parasitological diagnoses in naturally infected foxes. The objectives of this study were: first, to study the lobar and airway distribution of C. vulpis and E. aerophilus in wild red foxes and second, to investigate the relationship between fecal and histopathological diagnoses. Samples from 6 sites of the lung and fecal contents were obtained from 51 wild foxes in Prince Edward Island. By fecal examination, 78.4% of wild foxes tested positive for C. vulpis and 68.6% for E. aerophilus. In contrast, 66.6% and 49% of foxes had histopathological evidence of C. vulpis and E. aerophilus in the lungs, respectively. Anatomically, C. vulpis was observed in the small bronchi and bronchioles of all pulmonary lobes whereas E. aerophilus was restricted to the large bronchi and the caudal lobes. Affected airways exhibited severe epithelial glandular hyperplasia and bronchiolar mucous metaplasia. It was concluded that C. vulpis is widely distributed in airways of all pulmonary lobes, whereas E. aerophilus is mainly restricted to the bronchi of caudal lobes. Also, this study showed that histological examination of lung underestimates the infection with E. aerophilus.     

Potential application of serological tests on fluids from carcasses: detection of antibodies against Toxoplasma gondii and Sarcoptes scabiei in red foxes (Vulpes vulpes)

Background

Serological surveys for disease investigation of wild animal populations require obtaining blood samples for analysis, which has logistic, ethic and economic difficulties. Applying serological test to fluids collected from dead animals is an alternative. The aim of this study was to assess if antibodies could be detected in two types of fluids collected from 56 carcasses of red foxes (Vulpes vulpes): pleural fluid and lung extract.

Findings

In 22 (39%) foxes antibodies against Sarcoptes scabiei were detected in both fluid types by ELISA and Western blot. In 46 (82%) foxes, antibodies against Toxoplasma gondii were detected in pleural fluid and in 41 (73%) in lung extract applying a Toxo-screen test (DAT). Antibodies were still detectable in the same fluids kept at room temperature for 28 days, although in fewer foxes (16 and 14 foxes tested for T. gondii in lung extract and pleural fluid respectively; and 1 and 4 tested for S. scabiei in lung extract and pleural fluid respectively.

Conclusions

These results indicate the potential utility of using fluids from carcasses for antibody screening of wild animals at the population level.     

Experimental infection of red foxes (Vulpes vulpes) with Sarcoptes scabiei var. vulpes

Two of 3 red foxes became experimentally infected with Sarcoptes scabiei v. vulpes isolated from a naturally infected wild fox. The first clinical sign was seen after 10 days. After 30 days hairless patches were observed on the back of one of the foxes while general hair loss continued slowly. On the other fox the only macroscopical changes was tangling of the hair. The foxes died after 67 and 94 days, respectively. Both foxes were anaemic and in very poor condition. Scratching was seen in only 1 of the foxes.     

Rickettsiae in arthropods collected from red foxes (Vulpes vulpes) in France

The aim of our study was to detect the presence of Rickettsia spp. and Bartonella spp. in ticks and fleas collected from red foxes (Vulpes vulpes) in southeastern France during 2008. Using a genus-specific quantitative PCR (qPCR) assay, which was followed by a species-specific qPCR assay for the positive samples, 45.2% (33/73) of ticks (Rhipicephalus turanicus) were found to be infected with Rickettsia massiliae. 10.5% (2/19) of the fleas (Archaeopsylla erinacei) collected in the study tested positive for Rickettsia felis. A genus-specific qPCR assay did not reveal any Bartonella species in any of the ticks or fleas collected. The role of red foxes in the epidemiology of spotted fever caused by Rickettsiae species requires further investigation.     

Studies on the role of the red fox (Vulpes vulpes) as a potential definitive host of Neospora caninum

Neospora (N.) caninum is a protozoan parasite which is regarded as a major cause of abortion in cattle. Dogs and coyotes are definitive hosts of N. caninum which may shed environmentally resistant stages, oocysts, in their feces. Epidemiological studies in Germany showed that the presence of dogs increased the risk of a bovine herd to be N. caninum-positive in a bulk-milk ELISA test. However, there were also N. caninum-positive herds where dogs were not kept together with cattle.This leads to the question whether canids other than dogs, e.g., foxes, might be involved in the horizontal transmission of N. caninum. Therefore, the aim of our examinations in wild animals was to find out whether there are indications for a sylvatic cycle with foxes as definitive hosts and deer, roe deer and wild mice samples contained structures which resembled those of coccidian oocysts. In 13 of these 65 samples coccidian DNA was detected using a 18S rRNA gene based polymerase chain reaction (PCR).The examination of the 65 samples in a N. caninum-specific PCR revealed no positive result. Hammondia (H.) heydorni-DNA was detected in two samples. In addition, brain samples from 528 foxes, 224 wild mice, 16 deer and roe deer as well as from 1 wild boar were examined for the presence of N. caninum DNA by real time PCR. All samples tested negative by PCR. In conclusion, our study yielded no evidence indicating that the examined animals were part of a sylvatic cycle for N. caninum.