Neospora caninum Infection in a Litter of Labrador Retriever Dogs in Denmark

Neospora caninum is a recently recognized cyst-forming coccidian parasite associated with severe encephalomyelitis and myositis in dogs of different breeds and ages (Bjerkås et al 1984, Bjerkås & Presthus 1988, Dubey et al. 1988), but has for many years been misdiagnosed as Toxoplasma gondii. In some dogs, the main clinical sign has been attributed to polyradiculoneuritis (Dubey et al. 1988, Cuddon et al. 1992). Furthermore, ulcerative dermatitis (Dubey et al. 1988) and megaoesophagus have been reported (Wolf et al. 1991). The life cycle of the parasite and mode of infection have not been clarified, but transplacental infection seems so far to be the natural route of transmission between intermediate hosts (Dubey & Lindsay 1989). It has been speculated that the disease in young and adult dogs might be due to reactivation of a persistent infection because corticosteroid therapy can activate a latent N. caninum infection (Dubey & Lindsay 1993).     

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).     

Neosporosis

Neospora caninum is a recently recognized protozoan parasite of animals, which, until 1988, was misdiagnosed as Toxoplasma gondii. Neospora caninum or Neospora-like parasites, here discussed by J.P. Dubey and David Lindsay, cause paralysis and death in dogs and neonatal mortality and abortion in cattle, sheep, goats and horses.     

A Case of Neospora Associated Bovine Abortion in Sweden

Neospora caninum is a recently recognized protozoan organism that causes fatal neuromuscular disease in dogs and abortions and stillbirths in cattle and other animals (Dubey & Lindsay 1993). The parasite is morphologically similar and phylogenetically very closely related to the cyst-forming coccidium Toxoplasma gondii (Ellis et al. 1994, Holmdahl et al. 1994). This group of parasites has a two-host life cycle principally involving a carnivorous definitive host and a herbivorous or omnivorous intermediate host. However, with N. caninum, there is as yet no knowledge of any definitive host harbouring sexual stages of the parasite. The only known route of transmission is vertical from mother to foetus (Dubey & Lindsay 1993).     

Toxoplasma gondii infection in horses. A review

This review updates those written by Dubey and Beattie in 1988 (1988a) and by Tenter et al in 2000, on pathological and epidemiological aspects of Toxoplasma infection in horses. Under natural conditions, seroprevalence may variate from 0% up to 90%. This wide variation may be due to the sensitivity of the serological methods, to the age of animals, to the geographical area, and even to the hygienic condition of the farms and farm management. With few exceptions, horses are considered one of the less sensitive specie to the pathogenic effect of Toxoplasma gondii. In fact, neither under experimental nor under natural condition a genuine pathologic picture related to the toxoplasmic infection has been described. In one occasion the organism has been isolated from an eye condition and in others a connection between a higher frequency of unspecified pathological conditions and a positive response to serological test for Toxoplasma has been speculated. Diaplacental transmission and the following abortion have been only occasionally reported, and at least in one case in a quite trustworthy way, therefore it must be considered possible, though rare. Although infection of humans due to the consumption of horse meat has never been reported, the existence of a possible risk arouses by the demonstration of the presence of parasite stages in either naturally or experimentally infected horses, which resulted to be infective for mice and/or cats.     

Fatal toxoplasmosis in free-ranging endangered 'Alala from Hawaii

The 'Alala (Corvus hawaiiensis) is the most endangered corvid in the world, and intensive efforts are being made to reintroduce it to its former native range in Hawaii. We diagnosed Toxoplasma gondii infection in five free-ranging 'Alala. One 'Alala, recaptured from the wild because it was underweight and depressed, was treated with diclazuril (10 mg/kg) orally for 10 days. Antibodies were measured before and after treatment by the modified agglutination test (MAT) using whole T. gondii tachyzoites fixed in formalin and mercaptoethanol. The MAT titer decreased four-fold from an initial titer of 1:1,600 with remarkable improvement in physical condition. Lesions of toxoplasmosis also were seen in two partially scavenged carcasses and in a third fresh intact carcass. Toxoplasma gondii was confirmed immunohistochemically by using anti-T. gondii specific serum. The organism was also cultured by bioassay in mice from tissues of one of these birds and the brain of a fifth 'Alala that did not exhibit lesions. The life cycle of the parasite was experimentally completed in cats. This is the first record of toxoplasmosis in 'Alala, and the parasite appears to pose a significant threat and management challenge to reintroduction programs for 'Alala in Hawaii.     

The History of Toxoplasma gondii—The First 100 Years

ABSTRACT. In this paper the history of Toxoplasma gondii and toxoplasmosis is reviewed. This protozoan parasite was first discovered in 1908 and named a year later. Its medical importance remained unknown until 1939 when T. gondii was identified in tissues of a congenitally infected infant, and veterinary importance became known when it was found to cause abortion storms in sheep in 1957. The discovery of a T. gondii specific antibody test, Sabin–Feldman dye test in 1948 led to the recognition that T. gondii is a common parasite of warm‐blooded hosts with a worldwide distribution. Its life cycle was not discovered until 1970 when it was found that felids are its definitive host and an environmentally resistant stage (oocyst) is excreted in feces of infected cats. The recent discovery of its common infection in certain marine wildlife (sea otters) indicates contamination of our seas with T. gondii oocysts washed from land. Hygeine remains the best preventive measure because currently there is no vaccine to prevent toxoplasmosis in humans.     

Investigation of anti-Toxoplasma gondii antibodies in cats of the Ankara region of Turkey Using the Sabin-Feldman dye test and an indirect fluorescent antibody test

Blood samples from 99 cats from the Ankara province of Turkey were examined for the presence of anti-Toxoplasma gondii antibody with the use of both the Sabin-Feldman dye test (DT) and an indirect fluorescent antibody test (IFAT). Forty of the 99 sera (40.3%) were positive for antibodies against T. gondii with the DT, whereas the IFAT assay detected antibodies in 34 (34.3%). The study also evaluated 3 factors for their potential association with the presence of T. gondii antibody: age (<1 yr, 1-2 yr, and >2 yr), gender (female vs. male), and outdoor access (stray, owned with outdoor access, or indoor only). The DT detected antibodies in 3 cats under 1 yr of age, 22 cats between 1 and 2 yr, and 15 cats older than 2 yr, whereas the IFAT found 1, 18, and 15 cats positive for antibodies, respectively, in each of these categories. Of 61 female cats, 27 (44.2%) were positive by the DT; and of 38 male cats, 13 (34%) were positive by the DT. For the IFAT, 24 female cats (39.3%) and 10 male cats (26.3%) were positive. The percent seropositivity in indoor cats was 30.8% by the DT and 23.1% by the IFAT. In stray cats, the percent seropositivity was 52.8% by the DT and 41.7% by the IFAT. Antibody presence was significantly associated with age, but not with outdoor access.     

宠物弓形虫的感染及其防控

弓形虫病是一种世界性分布的人兽共患寄生虫病,对人类,尤其是妇女、儿童危害很大,估计全世界有1/3人受到该病的威胁。孕妇感染弓形虫后导致早产、流产、胎儿发育畸形;弓形虫是免疫功能低下患者的主要死亡原因之一。犬、猫是弓形虫的中间宿主和终末宿主,是人类感染弓形虫的主要来源。随着我国经济的迅速发展和人民生活水平的不断提高,城市中饲养犬、猫作为宠物的人越来越多,人、宠物间的亲密接触增加了弓形虫病传播给人的机会。加强对宠物犬、猫弓形虫病的研究及防控势在必行。本文就弓形虫的危害、宠物犬猫弓形虫感染及其防控措施作以综述。     

Evaluation of two Toxoplasma gondii serologic tests used in a serosurvey of domestic cats in California

We evaluated the sensitivity (Se) and specificity (Sp) of an IgG enzyme-linked immunosorbent assay (ELISA) and IgG indirect fluorescent antibody test (IFAT) for detection of Toxoplasma gondii-specific antibodies in sera from 2 cat populations using a Bayesian approach. Accounting for test covariance, the Se and Sp of the IgG ELISA were estimated to be 92.6% and 96.5%, and those of the IgG IFAT were 81.0% and 93.8%, respectively. Both tests performed poorly in cats experimentally coinfected with feline immunodeficiency virus and T. gondii. Excluding this group, Se and Sp of the ELISA were virtually unchanged (92.3% and 96.4%, respectively), whereas the IFAT Se improved to 94.2% and Sp remained stable at 93.7%. These tests and an IgM ELISA were applied to 123 cat sera from the Morro Bay area, California, where high morbidity and mortality attributable to toxoplasmosis have been detected in southern sea otters. Age-adjusted IgG seroprevalence in this population was estimated to be 29.6%, and it did not differ between owned and unowned cats. Accounting for Se, Sp, and test covariances, age-adjusted seroprevalence was 45.0%. The odds for T. gondii seropositivity were 12.3-fold higher for cats aged >12 mo compared with cats aged <6 mo.     

Toxoplasmosis: the first commercial vaccine

Infection with Toxoplasma gondii can have serious consequences in pregnant women and in immunocompromised individuals. For the farming industry it is a major cause of economic loss through neonatal mortality, particularly in sheep. In the following short review, David Buxton summarizes the main features of the complex immune response to the parasite and outlines how the first commercial vaccine was developed to control toxoplasmosis in pregnant sheep. Although the vaccine is inappropriate for use in human beings, the manner in which it induces immunity in sheep will prove vital to our understanding of the infection and the eventual development of a suitable vaccine to combat toxoplasmosis in people as well as in animals.     

Age-dependent impairment of functional helper T cell responses to immunodominant epitopes of Toxoplasma gondii antigens in congenitally infected individuals

Human infection with Toxoplasma gondii is generally asymptomatic in immunocompetent adults while it causes significant morbidity in congenitally infected children. Cell mediated immunity plays the main role in host resistance to T. gondii infection and a Th1 cytokine profile is necessary for protection and control of infection. The present work focused on comparing the helper T cell response to the GRA1 antigen of the parasite between children with congenital toxoplasmosis and healthy adults with acquired infection. We demonstrated that in young children with congenital infection the specific T cell response to parasite antigens is impaired and that such hypo-responsiveness is restored during childhood. Also, we provided clear evidence that in individuals with congenital toxoplasmosis the acquisition of functional helper T cell responses is disease-unrelated and indistinguishable in terms of strength, epitope specificity, and cytokine profile from the corresponding responses in immunocompetent adults with asymptomatic acquired T. gondii infection.     

Frequency and risk factors for toxoplasmosis in ovines of various regions of the State of Colima, Mexico

Infection of ewes by Toxoplasma gondii may induce abortions, thus it has consequences for sheep production. Colima, Western State of Mexico, has favourable climatic conditions for transmission and both domestic and wild cats live there. The aim of this study was to determine the frequency of specific antibodies in sheep from the coast, a mountain, and a hill of Colima. Serum samples from 351 sheep were tested by a previously standardized indirect ELISA. The frequency of infection was estimated and the farm location and flock size, as well as the animals' age and sex were analysed as risk factors for toxoplasmosis. The frequency of antibodies depended on the altitude, being higher at sea level than at 1200 metres above sea level (OR=3.77, 95% CI=1.79-7.94, P<0.0001), and the size of the flock, being higher in the large ones (OR=2.23, 95% CI=1.35-3.71, P=0.002). Older animals were more frequently positive and with a stronger response than young ones (OR=1.77, 95% CI=1.07-2.93, P=0.016). No differences were observed between male and female sheep. In conclusion, toxoplasmosis is present in sheep of Colima with variations related to altitude, flock size and age.     

Evaluation of an indirect fluorescent antibody test (IFAT) for demonstration of antibodies to Toxoplasma gondii in the sea otter (Enhydra lutris)

An indirect fluorescent antibody test (IFAT) for detection of Toxoplasma gondii infection was validated using serum from 77 necropsied southern sea otters (Enhydra lutris nereis) whose T. gondii infection status was determined through immunohistochemistry and parasite isolation in cell culture. Twenty-eight otters (36%) were positive for T. gondii by immunohistochemistry or parasite isolation or both, whereas 49 (64%) were negative by both tests. At a cutoff of 1:320, combined values for IFAT sensitivity and specificity were maximized at 96.4 and 67.3%, respectively. The area under the receiver-operating characteristic curve for the IFAT was 0.84. A titer of 1:320 was used as cutoff when screening serum collected from live-sampled sea otters from California (n = 80), Washington (n = 21), and Alaska (n = 65) for T. gondii infection. Thirty-six percent (29 out of 80) of California sea otters (E. lutris nereis) and 38% (8 out of 21) of Washington sea otters (E. lutris kenyoni) were seropositive for T. gondii, compared with 0% (0 out of 65) of Alaskan sea otters (E. lutris kenyoni).     

A chloroplast‐derived Toxoplasma gondii GRA4 antigen used as an oral vaccine protects against toxoplasmosis in mice

The parasitic protozoan Toxoplasma gondii, the causal agent of toxoplasmosis, can infect most mammals and birds. In human medicine, T. gondii can cause complications in pregnant women and immunodeficient individuals, while in veterinary medicine, T. gondii infection has economic importance due to abortion and neonatal loss in livestock. Thus, the development of an effective anti‐Toxoplasma vaccine would be of great value. In this study, we analysed the expression of T. gondii GRA4 antigen by chloroplast transformation (chlGRA4) in tobacco plants and evaluated the humoral and cellular responses and the grade of protection after oral administration of chlGRA4 in a murine model. The Western blot analysis revealed a specific 34‐kDa band mainly present in the insoluble fractions. The chlGRA4 accumulation levels were approximately 6 μg/g of fresh weight (equivalent to 0.2% of total protein). Oral immunization with chlGRA4 resulted in a decrease of 59% in the brain cyst load of mice compared to control mice. ChlGRA4 immunization elicited both a mucosal immune response characterized by the production of specific IgA, and IFN‐γ, IL‐4 and IL‐10 secretion by mesenteric lymph node cells, and a systemic response in terms of GRA4‐specific serum antibodies and secretion of IFN‐γ, IL‐4 and IL‐10 by splenocytes. Our results indicate that oral administration of chlGRA4 promotes the elicitation of both mucosal and systemic balanced Th1/Th2 responses that control Toxoplasma infection, reducing parasite loads.     

Exploring the ecologic basis for extreme susceptibility of Pallas' cats (Otocolobus manul) to fatal toxoplasmosis

Recent efforts by North American zoos to establish a genetically viable captive population of Pallas' cats (Otocolobus manul) have been compromised by high newborn mortality (approximately 60%), primarily because of toxoplasmosis. The basis for this extreme susceptibility to toxoplasmosis is unknown. In the present study, the general health status of wild Pallas' cats in Mongolia was evaluated, including assessment of basal hematologic parameters and fecal corticoid metabolite concentrations. The prevalence of exposure to Toxoplasma gondii in Mongolian Pallas' cats, local domestic cats, and prey species also was determined based on serology and/or polymerase chain reaction analysis. Biologic samples (blood, feces, and/or brain tissue) were obtained from 15 wild Pallas' cats, 15 domestic cats, and 45 prey animals (rodents and pikas) captured in Mongolia during the summers of 2000 and 2001. Comparative data were obtained from nine captive Pallas' cats maintained in North American zoos. Based on physical examinations, complete blood counts, and blood chemistry analyses, only minor differences were observed in the general health status of wild and captive Pallas' cats. Fecal cortisol metabolite concentrations did not differ (P > 0.05) between populations, indicating that Pallas' cats in captivity and in the wild have similar basal adrenocortical activity. A pronounced difference (P < 0.01) in seroprevalence to T. gondii was observed between populations. Whereas all captive Pallas' cats exhibited elevated immunoglobulin titers (IgG > 2,048) to T. gondii, only two of 15 (13%) wild Pallas' cats were seropositive, with both cats having lower IgG titers (< 1,024). Furthermore, no evidence of exposure to this parasite was found in any of the Mongolian domestic cats or prey species. These findings suggest that wild Pallas' cats have minimal opportunity for exposure to T. gondii in their natural habitat and, typically, do not become infected with this parasite until being brought into captivity. Accordingly, maintenance of a viable captive population may require implementing effective strategies to prevent exposure of immunologically naive Pallas' cats to T. gondii and to reduce parasite transmission between seropositive females and their highly susceptible offspring.     

Sero-epidemiological update on sheep toxoplasmosis in Sardinia, Italy

In order to better understand the epidemiology of ovine toxoplasmosis in Sardinia, a serological survey was carried out on 22 flocks with no fertility problems. In total 1043 sera (9% of the 11,382 sheep raised in the flocks) were examined by means of a commercial ELISA kit. To verify the performance of ELISA test, 160 selected sera were tested again with a gold standard test (IFAT). Performance of the commercial ELISA kit was summarised in terms of Sensitivity (SE), Specificity (SP), positive and negative Likelihood Ratios (LR+; LR-). The overall seroprevalence with ELISA test was recorded as 51.3%. It was generally increasing according to age and was significantly lower in animals younger than one year (with the exception of < 1 month old lambs). This survey provided data on the current Toxoplasma gondii sheep seroprevalence in Sardinia, confirmed a still high parasite pressure and pointed out that consumption of raw or undercooked ovine meat can be considered a potential risk factor for humans.     

Prevalence of antibodies to Toxoplasma gondii in horses in Riyadh Province, Saudi Arabia

The aim of the present study was to determine the seroprevalence of Toxoplasma gondii in horses used for sporting purposes in the Province of Riyadh, Saudi Arabia. In total, 266 serum samples from clinically healthy horses were analyzed for anti- T. gondii antibodies using the Sabin-Feldman dye test. Antibodies to T. gondii were found in 84 (31.6%) horses, with specific titers of 1∶16 (78 with a prevalence of 29.3%), 1∶64 (4 with a prevalence of 1.5%), and 1∶256 (2 with a prevalence of 0.8%). The number of seropositive horses in Shaqra (43.7%) was considerably higher than in other regions, with the lowest occurring in Al-Majmaah (11.0%). However, the differences in seroprevalence between the 6 locations were not statistically significant (P > 0.05). In contrast, the overall seroprevalence in Riyadh province was higher than that reported in other countries. Horses are not a direct source of infection for human toxoplasmosis in this region. Usually human infection is facilitated via cats, which are the reservoir hosts for this parasite.     

Seroprevalence and isolation of Toxoplasma gondii from sheep from São Paulo state, Brazil

Sheep are important in the epidemiology of Toxoplasma gondii infection but little is known of ovine toxoplasmosis in Brazil. Antibodies to T. gondii were assayed in sera of 495 sheep from 36 counties of S?o Paulo State, Brazil, using the modified agglutination test (MAT titer > or =1:25) and found in 120 (24.2%). Samples of brain, heart, and diaphragm of 82 seropositive sheep were pooled, digested in pepsin, and bioassayed in mice. Toxoplasma gondii was isolated from tissue homogenates of 16 sheep and the isolates were designated TgShBr1-16. Six of the 16 T. gondii isolates killed 100% of infected mice. Results indicate that asymptomatic sheep can harbor mouse-virulent T. gondii, and hence they can serve as a source of infection for humans.     

Prevalence of Toxoplasma gondii in stray cats of Gyeonggi-do, Korea

Toxoplasma gondii is an obligate intracellular zoonotic protozoan with a worldwide distribution. It infects humans as well as a broad spectrum of vertebrate hosts. Cats and wild felidae play crucial roles in the epidemiology of toxoplasmosis. This study was performed to survey the prevalence of T. gondii infection among stray cats in the Gyeonggi-do, Republic of Korea. A total of 174 stray cat blood samples were collected from Gwacheon-si (n=20), Bucheon-si (82), and Yangju-si (72). Positive sera for T. gondii were identified in 14 samples (8.1%) exclusively via the latex agglutination test, 28 (16.1%) via ELISA, and 23 (13.2%) via PCR analysis. The overall infection rate of female stray cats (29.2%) presented as higher than that of male cats (24.0%). This study suggests that T. gondii is widespread in the stray cat population of Gyeonggi-do, Korea. It is urgently needed to control urban stray cat population and to reduce the risk of zoonotic transmission of toxoplasmosis to other animal hosts and humans.