Genetic characterisation of Toxoplasma gondii in wildlife from North America revealed widespread and high prevalence of the fourth clonal type.

Little is known of the genetic diversity of Toxoplasma gondii circulating in wildlife. In the present study wild animals, from the USA were examined for T. gondii infection. Tissues of naturally exposed animals were bioassayed in mice for isolation of viable parasites. Viable T. gondii was isolated from 31 animals including, to our knowledge for the first time, from a bald eagle (Haliaeetus leucocephalus), five gray wolves (Canis lupus), a woodrat (Neotoma micropus), and five Arctic foxes (Alopex lagopus). Additionally, 66 T. gondii isolates obtained previously, but not genetically characterised, were revived in mice. Toxoplasma gondii DNA isolated from these 97 samples (31+66) was characterised using 11 PCR-restriction fragment length polymorphism (RFLP) markers (SAG1, 5'- and 3'-SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). A total of 95 isolates were successfully genotyped. In addition to clonal Types II, and III, 12 different genotypes were found. These genotype data were combined with 74 T. gondii isolates previously characterised from wildlife from North America and a composite data set of 169 isolates comprised 22 genotypes, including clonal Types II, III and 20 atypical genotypes. Phylogenetic network analysis showed limited diversity with dominance of a recently designated fourth clonal type (Type 12) in North America, followed by the Type II and III lineages. These three major lineages together accounted for 85% of strains in North America. The Type 12 lineage includes previously identified Type A and X strains from sea otters. This study revealed that the Type 12 lineage accounts for 46.7% (79/169) of isolates and is dominant in wildlife of North America. No clonal Type I strain was identified among these wildlife isolates. These results suggest that T. gondii strains in wildlife from North America have limited diversity, with the occurrence of only a few major clonal types.

Serological and parasitological prevalence of Toxoplasma gondii in wild birds from Colorado.

Ground-feeding birds are considered important in the epidemiology of Toxoplasma gondii because they serve as indicators of soil contamination by oocysts, and birds of prey are indicators of T. gondii prevalence in rodents and other small mammals. Cats excrete environmentally resistant oocysts after consuming tissues of T. gondii -infected birds. In the present study, sera and tissues from 382 wild birds from Colorado were tested for T. gondii infection. Antibodies to T. gondii were found in 38 birds with the use of the modified agglutination test (MAT, 1∶25 titer). Tissues (brains, hearts) of 84 birds were bioassayed in mice. Viable T. gondii was isolated from 1 of 1 barn owl (Tyto alba), 1 of 5 American kestrels (Falco sparverius), 1 of 7 ferruginous hawks (Buteo regalis), 1 of 4 rough-legged hawks (Buteo lagopus), 2 of 13 Swainson's hawks (Buteo swainsoni), and 1 of 25 red-tailed hawks (Buteo jamaicensis). This is the first time T. gondii has been isolated from the barn owl, ferruginous hawk, rough-legged hawk, and Swainson's hawk.

Morphologic and molecular characterization of the sarcocysts of Sarcocystis rileyi (Apicomplexa: Sarcocystidae) from the mallard duck ( Anas platyrhynchos ).

Macroscopic sarcocysts are often observed in ducks, but at present their taxonomic status remains uncertain because ducks serve as intermediate hosts for several such parasites in the genus Sarcocystis . One such species, Sarcocystis rileyi , was long ago established to involve the northern shoveler duck ( Anas clypeata ) and the striped skunk ( Mephitis mephitis ) as its intermediate and definitive hosts, respectively. Here, we employed light microscopy, electron microscopy, and DNA sequencing to more precisely describe diagnostic attributes of parasites presumed to represent S. rileyi occurring in a naturally-infected mallard duck ( Anas platyrhynchos ). By light and transmission electron microscopy, sarcocysts from the mallard duck resembled the S. rileyi described from A. clypeata . We document 18S, ITS-1, and 28S rDNA sequences from the mallard duck, the first for S. rileyi from any host. Sequences of conserved and variable portions of nuclear ribosomal DNA indicated that S. rileyi is related to, but distinct from, parasites employing opossums as their definitive host (including Sarcocystis neurona and Sarcocystis falcatula ). Diagnostic ultrastructural features and nucleotide sequences should aid in future studies and communications regarding this parasitic taxon, which lends itself to experimentation because its sarcocysts are macroscopic and easily excised from infected birds.

Effects of trapping and subsequent short-term confinement stress on plasma corticosterone in the brown treesnake (Boiga irregularis) on Guam.

The effects of capture in a live trap and subsequent handling stress on plasma concentrations of corticosterone and other sex steroids were examined in wild male and female brown treesnakes (Boiga irregularis), an introduced species on Guam that has been implicated in the extirpation or decline of many of that island's vertebrate species. Males and females that spent 1 night in a trap had plasma levels of corticosterone about four and two times higher, respectively, than those of the respective free-ranging controls. Mean plasma levels of corticosterone of snakes that had spent 3 nights in a trap were intermediate between, but not significantly different from, those of snakes that had spent 1 night in a trap and free-ranging snakes, suggesting that some acclimation to capture occurred during this period. Snakes that were taken from traps and held in collecting bags for 10 min and 2 h prior to blood sampling had levels of corticosterone about two and three times higher, respectively, than those of control snakes that were taken from traps and bled immediately. Concentrations of plasma corticosterone in free-ranging females were about two times higher than those of males but were well within the range of basal levels observed in other reptiles. Few snakes of potential reproductive size were reproductive (males: 1 of 35; females: 2 of 33), and plasma concentrations of testosterone and progesterone in nonreproductive males and females, respectively, were accordingly low. The possible relationship between corticosterone and these sex steroids, therefore, could not be adequately assessed, although there was a positive relationship between plasma progesterone and corticosterone in the nonreproductive females. Nonetheless, as a prerequisite for studies on the seasonal hormonal cycles of this species on Guam, our observations raise the possibility that the stress caused by trapping could affect the levels of other sex steroids and that, therefore, such studies should use free-ranging individuals.