Factors affecting seroprevalence of Toxoplasma gondii in the endangered Iberian lynx (Lynx pardinus)

Wild felids are considered important in maintaining the sylvatic cycle of Toxoplasma gondii. Although, T. gondii antibodies have been reported in several species of wild felids, little is known of the epidemiology and risk factors associated with T. gondii infection in wild cats. The Iberian lynx (Lynx pardinus) is the most endangered felid species in the world. In the present study, seroprevalence and associated risk factors for T. gondii infection in a large population of Iberian lynx in Spain were determined. Serum samples from 129 Iberian lynx collected from 2005 to 2009 and 85 wild rabbits (Oryctolagus cuniculus), sharing the habitat with the Iberian lynx, were tested for antibodies to T. gondii by the modified agglutination test (MAT) using a cut-off value of 1:25. Antibodies to T. gondii were found in 81 of 129 (62.8%) Iberian lynx. Seroprevalence to T. gondii in Iberian lynx significantly increased with age (P < 0.001). T. gondii seroprevalences were similar in free-ranging (66.7% of 93) and wild-caught captive lynx (69% of 84) but significantly lower in captive-born lynx (22.5% of 40). Seroprevalence was higher in lynx with concurrent Cytauxzoonfelis (88% of 25) but not with concurrent Feline Leukemia Virus (FeLV) infection (53.8% of 13). There were no significant differences in seroprevalence between sexes, geographic region and year of sample collection (2005–2009). Oocysts of T. gondii were not detected microscopically in fecal samples from 58 lynx. Wild rabbits are considered the most important food for the lynx. Antibodies to T. gondii were found in 14 (11.9%) of 85 rabbits tested. The present results indicate that T. gondii infection is widespread in the two areas where Iberian lynx survive in Spain. The fact that four captive-born lynx seroconverted was indication of contact with T. gondii also in the Captive Breeding Centers, hence, control measures to prevent T. gondii infection would be necessary in these centers.     

Pathogenesis of acute toxoplasmosis in specific-pathogen-free cats

Systemic toxoplasmosis was produced in specific-pathogen-free cats by intravenous inoculation with Toxoplasma gondii tachyzoites. Infectious organisms were recovered from all tissues studied, but the number of organisms recovered from liver, lungs and spleen was 10-fold to 10,000-fold higher than from heart and brain. The occurrence and severity of Toxoplasma-induced lesions correlated with the number of infectious organisms recovered from the various tissues. In nonlymphoid tissues, the Toxoplasma-associated lesions consisted of multifocal necrosis, usually with demonstrable organisms. Lesions in the spleen and mesenteric lymph nodes consisted of reticuloendothelial and lymphoid hyperplasia, with few demonstrable organisms. Pneumonitis was severe and sometimes fatal in the early stages of systemic toxoplasmosis. Light- and electron-microscopic studies showed that the earliest lung lesions were randomly distributed infiltrates of neutrophils, eosinophils, and mononuclear cells into alveolar walls. Later lesions were diffuse alveolar necrosis, pneumocytic hyperplasia, and extensive fibrinocellular exudates in alveoli. Tachyzoites were present in cytoplasmic vacuoles of fibroblasts, macrophages, type I and II pneumocytes, bronchiolar epithelial cells, bronchiolar smooth muscle cells, endothelial cells, neutrophils, and eosinophils, and circulating monocytes. Replication of organisms was found in all parasitized cell types except neutrophils and eosinophils.     

Genetic and biologic characteristics of Toxoplasma gondii infections in free-range chickens from Austria

The prevalence of Toxoplasma gondii in free-ranging chickens is a good indicator of the prevalence of T. gondii oocysts in the soil because chickens feed from the ground. The prevalence of T. gondii in free-range chickens (Gallus domesticus) from 11 Bio-farms in Austria was determined. Antibodies to T. gondii assayed by the modified agglutination test (MAT) were found in 302 of 830 (36.3%) chickens with titers of 1:10 in 50, 1:20 in 69, 1:40 in 53, 1:80 in 40, 1:160 or higher in 90. Hearts of 218 chickens with MAT titers of 10 or higher were bioassayed individually in mice. Tissues from 1183 chickens were pooled and fed to 15, T. gondii-free cats. Feces of the cats were examined for oocysts; 11 cats shed T. gondii oocysts. T. gondii was isolated from 56 chickens by bioassay in mice. Thus, there were 67 isolates of T. gondii from these chickens. Genotyping of these 67 isolates using the SAG2 locus indicated that all 33 were Type II. Phenotypically and genetically these isolates were different from T. gondii isolates from Brazil. None of the isolates was virulent for mice. This is the first report of isolation of T. gondii from chickens from Austria.     

Myelitis in a cat infected with Toxoplasma gondii and feline immunodeficiency virus

Severe necrotizing myelitis secondary to localization and reactivation of Toxoplasma gondii within the spinal cord of a domestic shorthair cat was diagnosed by use of light and electron microscopy and immunohistochemistry. The cat also was infected with feline immunodeficiency virus. This case may have useful comparative features to T gondii infections in human patients with acquired immunodeficiency syndrome.     

Clinical Sarcocystis neurona, Sarcocystis canis, Toxoplasma gondii, and Neospora caninum infections in dogs

Sarcocystis neurona, Sarcocystis canis, Toxoplasma gondii, and Neospora caninum are related apicomplexans that can cause systemic illness in many species of animals, including dogs. We investigated one breeder's 25 Basset Hounds for these infections. In addition, tissues from dogs and other non-canine hosts previously reported as S. canis infections were studied retrospectively. Schizonts resembling those of S. neurona, and recognized by polyclonal rabbit anti-S. neurona antibodies, were found in six of eight retrospective cases, as well as in two additional dogs (one Basset Hound, one Springer Spaniel) not previously reported. S. neurona schizonts were found in several tissues including the central nervous system, lungs, and kidneys. Fatal toxoplasmosis was diagnosed in an adult dog, and neosporosis was diagnosed in an adult and a pup related to the one diagnosed with S. neurona. No serological reactivity to S. neurona antibodies occurred when S. canis-like liver schizonts were retrospectively assayed from two dogs, a dolphin, a sea lion, a horse, a chinchilla, a black or either of two polar bears. Sequencing conserved (18S) and variable (ITS-1) portions of nuclear ribosomal DNA isolated from the schizont-laden liver of a polar bear distinguished it from all previously characterized species of Sarcocystis. We take this genetic signature as provisionally representative of S. canis, an assumption that should be tested with future sequencing of similar liver infections in other mammalian hosts. These findings further extend the uncharacteristically broad intermediate host range for S. neurona, which also causes a neurologic disease in cats, mink, raccoons, skunks, Pacific harbor seals, ponies, zebras, lynxes, and sea otters. Further work is necessary to delineate the causative agent(s) of other cases of canine sarcocystosis, and in particular to specify the attributes of S. canis, which corresponds morphologically to infections reported from wide range of terrestrial and marine mammals.     

The SnSAG merozoite surface antigens of Sarcocystis neurona are expressed differentially during the bradyzoite and sporozoite life cycle stages

Sarcocystis neurona is a two-host coccidian parasite whose complex life cycle progresses through multiple developmental stages differing at morphological and molecular levels. The S. neurona merozoite surface is covered by multiple, related glycosylphosphatidylinositol-linked proteins, which are orthologous to the surface antigen (SAG)/SAG1-related sequence (SRS) gene family of Toxoplasma gondii. Expression of the SAG/SRS proteins in T. gondii and another related parasite Neospora caninum is life-cycle stage specific and seems necessary for parasite transmission and persistence of infection. In the present study, the expression of S. neurona merozoite surface antigens (SnSAGs) was evaluated in the sporozoite and bradyzoite stages. Western blot analysis was used to compare SnSAG expression in merozoites versus sporozoites, while immunocytochemistry was performed to examine expression of the SnSAGs in merozoites versus bradyzoites. These analyses revealed that SnSAG2, SnSAG3 and SnSAG4 are expressed in sporozoites, while SnSAG5 was appeared to be downregulated in this life cycle stage. In S. neurona bradyzoites, it was found that SnSAG2, SnSAG3, SnSAG4 and SnSAG5 were either absent or expression was greatly reduced. As shown for T. gondii, stage-specific expression of the SnSAGs may be important for the parasite to progress through its developmental stages and complete its life cycle successfully. Thus, it is possible that the SAG switching mechanism by these parasites could be exploited as a point of intervention. As well, the alterations in surface antigen expression during different life cycle stages may need to be considered when designing prospective approaches for protective vaccination.     

Diagnosis of bovine neosporosis

The protozoan parasite Neospora caninum is a major cause of abortion in cattle. The diagnosis of neosporosis-associated mortality and abortion in cattle is difficult. In the present paper we review histologic, serologic, immunohistochemical, and molecular methods for dignosis of bovine neosporosis. Although not a routine method of diagnosis, methods to isolate viable N. caninum from bovine tissues are also reviewed.