Seroprevalence of Toxoplasma gondii and Neospora caninum in dogs from Korea

Toxoplasma gondii and Neospora caninum are closely related protozoan parasites, they share many common hosts, and can cause neurological diseases in dogs. Dogs can have close contacts with humans and livestock and therefore they can act as reservoirs of these parasites. The aim of this study was to survey the seroprevalence of antibodies against T. gondii and N. caninum and their co-infection rate in dogs in Korea. In total, sera from 553 domestic dogs were collected from different breeds, sexes, and ages of dogs from nine provinces across the country of Korea during 2006 and 2007. The presence of antibodies against T. gondii and N. caninum was analyzed using the latex agglutination test (LAT) with a cut-off value of 1:32, and the indirect fluorescent antibody test (IFAT) using a serum titer of 1:100. In the total dog population, 71 (12.8%) dogs were positive for anti-T. gondii antibodies and only 20 (3.6%) were positive for anti-N. caninum antibodies. Relatively higher seropositive frequencies of antibodies against T. gondii (20.1%) and N. caninum (4.9%) were detected in the dog population from the Gyeonggi. A higher proportion of animals seropositive for anti-T. gondii antibodies was found in stray dog populations as compared to household dog populations: 18.5% (59/319) vs 5.1% (12/234), respectively. The Chi-square tests revealed significant differences in the seropositive frequencies of antibodies against T. gondii between stray and household dogs in the total population (p<0.0001), and in dogs from the Gyeonggi (p<0.01). No significant differences were observed for the presence of antibodies against T. gondii or N. caninum when compared across the sex or age (p>0.05). The first serological survey on antibodies against both T. gondii and N. caninum parasites across the entire country showed that co-infection was not common in these canine populations with a seropositive level of 0.72%. The significantly higher positive frequency of T. gondii antibodies in stray dogs in both, Gyeonggi and in the total dog populations suggests that further investigation on the seroprevalence of parasites should focus on stray dogs.     

Serological study of Neospora caninum infection in dogs and cattle from west of Iran

The aim of this study was to determine the seroprevalence of Neospora caninum infection in dogs and cattle from Hamedan province (West of Iran). Blood samples were collected from 1,046 cattle and 270 dogs in this area. Cattle and dog samples were tested and analyzed using ELISA and IFAT, respectively. IgG-antibodies to N. caninum were found in 27 and 17.4 % of dogs and cattle samples, respectively. In cattle study, The association between infection and type of cattle was statistically significant (P?=?0.004). Also, significant statistical differences were observed regarding to stray canids presence in farm (P?P?P?=?0.195) and breed (P?=?0.077). In dog study, there was statistical differences among age groups (P?P?P?=?0.112). This study is the first report of N. caninum infection in dogs and cattle from west of Iran. There is both horizontal and vertical transmission of N. caninum in this area, and the presence of stray dogs may be a risk factor for N. caninum infection in cattle. N. caninum is an important factor in the economic losses of the cattle breeding in Hamedan province. Therefore, further investigations and designing control strategies for improving management in cattle farms is highly recommended.     

Seroprevalence of Neospora caninum infection in dairy cows in Northern provinces,Thailand

Neospora caninum, an obligate intracellular protozoan parasite, is the causative agent of neosporosis, recognized as a major cause of bovine abortion around the world. Thailand is a developing agricultural country located in Southeast Asia. Livestock developments particularly in dairy cows of this country have been hampered by low productivity including milk and slow growth rate due to the impact of many pathogens including N. caninum. Currently, there is no effective method for control of neosporosis since there is less information regarding current status of infections. The objective of this study was to investigate the seroprevalence of neosporosis in dairy cows of the northern part of Thailand. During 2006–2007, the sera of 642 cows from 42 small farm holders with the top three highest consensus of dairy farms in the northern provinces, such as Chiang Rai, Chiang Mai and Lumpang were collected and performed tests. Antibodies to N. caninum were assayed by enzyme-linked immunosorbent assay (ELISA) with recombinant N. caninum surface antigen 1 (NcSAG1) and indirect fluorescent antibody test (IFAT). The overall prevalence of N. caninum infection in this study was 46.9% (301/642) by ELISA and 34.3% (220/642) by IFAT.     

Prevalence of antibodies to Neospora caninum in stray dogs of Urmia, Iran

To investigate anti-Neospora caninum antibodies in stray dogs living in Urmia city, 135 blood samples were collected. Serum samples were screened for detection of anti-N. caninum IgG antibodies using indirect fluorescent antibody test (IFAT; ≥50). Antibodies were seen in 36 (27%) of 135 dogs. The IFAT antibody titers were as follows: 1:50 in 16 dogs, 1:100 in ten dogs, 1:200 in six dogs, 1:400 in one dog, 1:800 in two dogs, and 1:1,600 in one dog. There were no significant differences in seroprevalence of Neospora infection between different genders (p > 0.05). The seropositive results were increased with age and the differences were statistically significant (p < 0.05). The results confirm the presence and exposure of stray dogs to N. caninum in Urmia city and the importance of this protozoan as a cause of disease in dogs of the region.     

Seroprevalence of Neospora caninum in racehorses in Ankara, Turkey

To assess seroprevalence of Neospora spp. in asymptomatic horses in Ankara, Turkey, 19 mares and 56 stallions older than 2 years of age were examined using ELISA; 9.3% of the horses were seropositive. Seroprevalence of N. caninum in mares was twice of that in stallions (15.8 vs 7.1%) and all appeared to be asymptomatic.     

Seroprevalence of Neospora caninum and Toxoplasma gondii in camels (Camelus dromedarius) in Mashhad, Iran

One hundred twenty camels were blood-sampled and used to evaluate serological screening for Neospora caninum and Toxoplasma gondii infection by indirect fluorescent antibody test (IFAT) in Mashhad, Iran, during years 2004–2005. Of the 120 camels, antibodies to N. caninum were found in three in titers of 1:20 and in four in titers of 1:40 using whole N. caninum tachyzoites as IFAT slide (VMRD Inc., Pullman, WA 99163, USA). Antibodies to T. gondii were found in three camels in titers 1:20 and in two camels in titers 1:40 using whole T. gondii tachyzoites as IFAT slide (BIOGENE, Iran).     

Seroprevalence of Neospora caninum in cats from the Czech Republic

Sera of 414 cats coming from different parts of the Czech Republic were tested for N. caninum antibodies. Sera samples were collected during years 2002–2011. N. caninum antibodies were detected by a commercial competitive-inhibition enzyme-linked immunosorbent assay (cELISA) with cut off ≥30% inhibition. Samples positive in cELISA were confirmed by an indirect fluorescence antibody test (IFAT); titre ≥50 was considered positive. In total, 137 (33%) cats reacted positively in cELISA; N. caninum antibodies in IFAT were detected in 16 (3.86%) cats with titres 50 and 100. In 6 cats, positive for N. caninum antibodies, T. gondii antibodies were also detected by IFAT. It is the first report of N. caninum antibodies in domestic cats from the Czech Republic and third report in Europe.     

Neospora caninum infection dynamics in dairy cattle

Neospora caninum is considered in many countries as one of the key abortion agents in cattle. This study aims to investigate the parasite behavior in dairy cattle in the municipality of Avaré, SP, Brazil, where abortions frequently occur. An ELISA was performed to compare two samplings on a total of 615 animals; tests were performed in the same herds with a gap of 3 years. An increase in the percentage of reactive animals was observed, ranging from 21.6% at the first sampling to 38.9% at the second sampling. Of the 176 animals tested at both samplings, 61.93% retained a non-reactive status, 15.9% retained a reactive status, 19.88% switched from non-reactive to reactive and 2.27% switched from reactive to non-reactive. Of the 100 animals with reproductive disturbances, 50% presented anti-Neospora antibodies, thereby indicating the presence of the protozoa. When comparing cows and their respective female offspring, a predominance of horizontal infection was observed. Moreover, considering the significant percentage of animals that switched from non-reactive to reactive and the abundant presence of dogs among the herds, the N. caninum transmission may be attributed to presence of carnivores.     

Seroprevalence of Toxoplasma gondii infection in shepherd dogs from Sarab District,Northwest Iran

The aim of this study was to compile initial epidemiological data on the prevalence of Toxoplasma gondii antibodies in Sarab area. We evaluated the titers of anti-T. gondii antibodies by latex agglutination test (LAT; titer?>?1:64) serological test in 39 (10.1 %) serum samples from shepherd dogs exhibiting clinical signs of infectious diseases. The largest age group was 2–4 years, with 170 (44.3 %) dogs out of which only 21 dogs (12.3 %) were seropositive and the lowest age group was 4–6 years, with 87(22.6 %) out of which only 3 dogs (3.4 %) were seropositive. The highest rate of infection was seen in Ardaha village, with 11 (27.5 %) dogs. The lowest rate of infection was detected in Khaki and Asbfroshan villages, with only one dog. According to dog breed, the seropositivity rate of T. gondii was 14.3 % in pure breed dogs and 11.2 % in mongrel dogs. No sex or breed predispositions to T. gondii infection were detected during this serological assays (P?≤?0.05). The antibody titres of T. gondii ranged from 1:64 to 1:2,084. 39 Seropositive dogs with titers of 1:64 in 10, 1:128 in 13, 1:256 in 5, 1:512 in 6, 1:1,024 in 3, and 1:2,084 in 2 dog. In the present study, there was no correlation between antibody titers and, age or location. Our results show that presence and exposure of shepherd dogs to T. gondii in Sarab city confirm that dogs are exposed to T. gondii and play an important role in the epidemiology of T. gondii of this region.     

Seroprevalence of <Emphasis Type="Italic">Neospora caninum</Emphasis> infection in dogs from rural and urban environments in Tehran,Iran

103 serum samples were taken from owned dogs in Tehran (53 urban dogs and 50 farm dogs) from February to May 2005 and tested for IgG antibody against Neospora caninum using the immunofluorescence antibody test. Urban dogs were selected from apparent healthy dogs referred to the Small Animal Hospital of Faculty of Veterinary Medicine, University of Tehran, for routine operation, and the farm dogs were selected from the sheepdogs in sheep flocks around Tehran randomly. The titer of 1:50 was selected as cutoff. In total, 20 of 103 cases (19.4%) were identified as positive. The infection rate in farm dogs (28%) was higher than in urban dogs (11.3%), and the difference was statistically significant (p < 0.05). The infection rates in different genders and age groups were determined. The differences between seroprevalence of N. caninum infection in different sexes were not significant. A high correlation was observed between the infection rate and age (r ² = 0.97, p < 0.01).     

Cyclic transmission of Neospora caninum: serological findings in dogs shedding oocysts

In a previous paper we demonstrated that Hammondia heydorni-like oocysts isolated in 1996 from a naturally infected dog could not be distinguished from the isolate Neospora caninum NC-1. The isolate, designated as H. heydorni-Berlin-1996, was cyclically transmitted using dogs as the final hosts. The present study provides information on the antibody responses of the dogs used for the cyclical transmission of this isolate. The majority of dogs that had shed oocysts showed no sero-conversion with respect to N. caninum tachyzoite surface or immunodominant antigens, either in the indirect fluorescent antibody test or in two Western-blot-based tests. In addition to the examination of responses to immunodominant antigens, we also analysed the antibody reactions of dogs to a high-molecular-weight antigen (152 kDa) in the tachyzoite antigen preparation. The antibodies against this antigen appeared after the dogs had been fed infected intermediate host tissues and shed oocysts. The reaction was observed in dogs between day 35 and day 447 after feeding of intermediate host tissues. Therefore, our study provides initial information on a 152 kDa tachyzoite antigen, which might be a suitable candidate to identify dogs with a history of shedding N. caninum oocysts.     

Direct agglutination test for serologic diagnosis of Neospora caninum infection

A direct agglutination test was evaluated for the detection and quantitation of IgG antibodies to Neospora caninum in both experimental and natural infections in various animal species. As compared with results obtained by the indirect fluorescent antibody test, the direct agglutination test appeared reliable for the serologic diagnosis of neosporosis in a variety of animal species. The direct agglutination test should provide easily available and inexpensive tools for serologic testing for antibodies to N. caninum in many host species. Received: 26 June 1997 / Accepted: 5 August 1997     

Molecular detection of Neospora caninum in house sparrows (Passer domesticus) in Iran

Neospora caninum is an intracellular protozoan parasite with a wide range of intermediate bird hosts. There is little information describing the prevalence and genetic characterization of N. caninum in bird hosts worldwide and in Iran. In this study, a total of 217 brain samples of house sparrow (Passer domesticus) were examined for N. caninum presence by nested polymerase chain reaction targeting the Nc-5 gene. N. caninum DNA was detected in 3.68% (8/217) of sparrows. Sequencing of the Nc5 genomic DNA revealed 97–99% of similarity with N. caninum sequences deposited in Genbank. To our knowledge, this study is the first molecular evidence of N. caninum DNA in bird hosts in Iran. The results of this study highlight the role of the house sparrow (Passer domesticus) in maintaining and spreading N. caninum infection to canines in the feral and domestic environment.     

Prevalence of antibodies against Neospora caninum in dogs from urban areas in Central Poland

Neospora caninum is a protozoan parasite which causes abortion in cattle as well as reproduction problems and neurological disorders in dogs. To assess the prevalence of the parasite in urban dogs in the Mazovian Voivodeship, Central Poland, serum samples from 257 dogs were analyzed for the presence of specific IgG antibodies. The examined dogs visited three private veterinary clinics located in Warsaw due to control tests, vaccinations, or other reasons not directly connected with neosporosis. Using ELISA and Western blot, antibodies against the parasite were detected in 56 out of 257 dogs, giving a prevalence of 21.7%. A greater prevalence was observed in female dogs than in males, 28% and 17.3%, respectively, and the differences were statistically significant (p < 0.05). There were no significant differences in seroprevalence of Neospora infection within the age groups (p > 0.05). This study indicates the presence of N. caninum in the Mazovian Voivodeship, in dogs which live in urban areas and exposure of these dogs to the parasite. The fact that seropositive dogs had no contact with cattle confirms the important role of dogs in the parasite’s epidemiology.     

Efficacy of toltrazuril and ponazuril against experimental Neospora caninum infection in mice

Neosporosis is a disease affecting predominantly fetal development in cattle and dog hosts; and it may cause neuromuscular disfunction in infected new-born calves and pups. Predispositions – including, e.g. transient immunosuppression during pregnancy – may result in an increased dissemination of the parasite within the host or its offspring. Chemotherapeutic treatment of neosporosis may be an issue, provided that an appropriate drug is made available. In this respect, we describe the use of a mouse model for the evaluation of toltrazuril and ponazuril medication as a means of preventing parasite dissemination and subsequent formation of cerebral lesions. Toltrazuril- and ponazuril-treated mice were experimentally infected intraperitoneally (i.p.) with 2 × 10⁶ Neospora caninum tachyzoites. The infection was monitored at three levels: clinically, by assessing symptoms, histologically, by assessing the occurrence of cerebral lesions and parasites by immunohistochemistry, and on the molecular level, by detection of parasite DNA using PCR. Chemotherapy using either toltrazuril or ponazuril, both applied in a drinking-water formulation (20 mg toltrazuril or ponazuril kg⁻¹ body weight day⁻¹) completely prevented the formation of cerebral lesions in all treated animals, as assessed by immunohistochemistry. PCR analyses of these treated animals showed that DNA-detectability was reduced by 91% and 90% upon toltrazuril and ponazuril medication, respectively. Received: 6 August 2000 / Accepted: 7 August 2000     

Kennel dogs as sentinels of Leishmania infantum, Toxoplasma gondii, and Neospora caninum in Majorca Island, Spain

Kennel dogs can serve as sentinels and/or reservoirs of diseases of veterinary and zoonotic interest because they have often roamed free and lived outdoors, thus being exposed to pathogens. We tested dogs from the kennel of Inca (Majorca/Mallorca, Balearic Islands, Spain) for evidence of infection with three protozoan parasites: Leishmania infantum, Toxoplasma gondii, and Neospora caninum. Exposure to L. infantum was found in 56.3% of 48 dogs (37.5% by Western blot, 43.8% by PCR). Only 30% of infected dogs had leishmaniosis-like lesions. Seroprevalence to T. gondii was 58.7% of 46 dogs using the modified agglutination test (MAT, titer 1:25). None of the 44 dogs tested had N. caninum antibodies using a commercial competitive ELISA, probably because the surveyed dogs did not roam in the proximity of cattle farms. Results confirm the endemicity of L. infantum and also the widespread presence of T. gondii in the Mediterranean island of Majorca.     

Stress-driven stage transformation of Neospora caninum

Neosporiosis, a serious disease caused by the apicomplexan intracellular parasite Neospora caninum, is considered to be one of the most economically important diseases in cattle. It is associated with potentially serious complications such as abortion, stillbirth and maternal infertility. To survive in fluctuating physiological and immunological environments, N. caninum has evolved a diverse set of regulatory mechanisms that govern various adaptive responses. The most intriguing paradigm in N. caninum adaptive evolutionary biology is its ability to alternate between two phenotypically and functionally distinct stages within the host. Recent research has reinforced the notion that N. caninum tachyzoite–bradyzoite stage switching on and off is correlated with its ability to form dormant cysts. Knowledge of the specific mechanisms that govern the dynamics of N. caninum phenotype switching enables a better understanding of the pathogenesis of the disease and effective control measures to be identified. Herein we review the available knowledge relating to various aspects of stage interconversion in N. caninum, with particular focus on the stress-related hypothesis presumed to be involved in this event. Finally, we put forward the postulation that N. caninum uses stage interconversion as an adaptive process to cope with the hostile environment within the host and to ensure its continuity in nature.     

Immune responses in pregnant cattle and bovine fetuses following experimental infection with Neospora caninum

Humoral and cell-mediated immune (CMI) responses [i.e. proliferative responses and gamma interferon (IFN-gamma) production], were elicited in five cows infected between 159 and 169 days of gestation by a combined intravenous-intramuscular inoculation of Neospora caninum tachyzoites. Analysis of antigen-specific immunoglobulin (IgG) subclasses revealed a predominant IgG2 response in two cows, a mixed IgG1-IgG2 response in two other cows and a predominant IgG1 response in one cow. No correlation was found between IgG2 titers and IFN-gamma levels. CD4-T cells were responsible for the CMI responses in peripheral blood mononuclear cells from three infected cows. All five fetuses removed from infected dams at week 9 post-infection (219-231 days of gestation) mounted strong Neospora-specific humoral responses and had a predominant IgG1 response, regardless of their ability to produce IFN-gamma. However, CMI responses were highly variable between fetuses. These data indicate the complexity of the immune mechanisms associated with Neospora infection in both the dams and their fetuses.     

Dense-granule protein NcGRA7, a new marker for the serodiagnosis of Neospora caninum infection in aborting cows

To investigate whether the production of an antigen-specific antibody is associated with Neospora caninum-induced bovine abortion, 62 serum samples were tested with an enzyme-linked immunosorbent assay using the recombinant antigens NcSAG1, NcSRS2, and NcGRA7. Our study suggested that NcGRA7 would be a new marker for the serodiagnosis of N. caninum infection resulting in abortion.