Disseminated Toxoplasma gondii Infection in an Adult Osprey (Pandion haliaetus)

An adult female osprey (Pandion haliaetus) was found weak and unable to fly in Auburn, Alabama in August 2019. The bird was captured and submitted to the Southeastern Raptor Center of the Auburn University College of Veterinary Medicine for evaluation. On presentation, the bird was thin with a body condition score of approximately 1.5 out of 5. The bird died during the examination and was submitted for necropsy. At the necropsy, there was a severe loss of muscle mass over the body, and the keel was prominent. The liver and spleen were moderately enlarged with pale tan to red foci randomly scattered throughout the parenchyma. A histopathologic observation revealed multifocal to coalescing areas of necrosis and hemorrhage with intralesional protozoans in the liver, spleen, lungs, kidney, sciatic nerve, esophagus, cerebrum, heart, and proventriculus. Immunohistochemistry using anti-Toxoplasma gondii-specific antibodies showed a strong positive labeling of the parasite. Semi-nested PCR, specific for the B1 gene of T. gondii, successfully identified T. gondii. This is the first confirmed case of T. gondii infection in an osprey.

Phylogenetic Analysis and Genetics Polymorphisms Evaluation of ROP8 and B1 Genes of Toxoplasma gondii in Livestock and Poul-try Hosts of Yazd, Qom and Golestan Provinces of Iran

Background: A high correlation is observed between specific clonal lineages and host types in toxoplasmosis. The main objectives of this study were comparing polymorphism and evolutionary analysis of the B1 and ROP8 genes, as well as the evaluation of phylogenic and Toxoplasma gondii isolates obtained from different hosts and regions. Methods: Overall 96 brain/ diaphragm tissue samples of livestock and poultry from three provinces of Iran (cows: 9 from Yazd, 9 from Qom; sheep: 19 from Yazd, 7 from Qom; goats: 7 from Yazd, 4 from Qom; one camel from Yazd and 37 chickens, 2 roosters and one duck from Golestan) were tested during 2018-19. A nested PCR and PCR-PCR methods were developed with the B1 and ROP8 genes. Evaluation of genetic proximity, genetic diversity and evolutionary analysis were done using MEGA-X and DnaSP5 software. Thirty samples of both genes were sequenced (18 B1 and 12 ROP8 genes), and submitted to the GenBank (MN275903-MN275932). Results: Tajima's D index analyses showed that both genes were in the negative direction of evolution. The B1 gene was more sensitive than the ROP8 gene. The ROP8 gene showed better and more acceptable results in terms of the relationship between the host and the genotyping of the samples. Conclusion: The B1 gene was only an attractive target for rapid detection of T. gondii parasites, whereas the ROP8 gene due to a high level of polymorphism was able to isolate the three clonal lineages (type I, II and III), inter-types and even atypical strains from different isolates of T. gondii.

Prevalence of Toxoplasma gondii infection among small mammals in Tatarstan, Russian Federation

Toxoplasma gondii is a zoonotic parasite with a wide host range that includes humans, domestic animals and wild animals. Small mammals serve as intermediate hosts for T. gondii and may contribute to the persistence of this parasite in the environment. Mass mortality in wild animals and deaths in rare endemic species make the study of this parasite of growing importance. In this study, T. gondii infection prevalence was evaluated in brain tissues from 474 small mammals captured at 26 trapping points in urban and rural areas of Tatarstan, Russian Federation. Nested PCR was used to detect the T. gondii B1 gene in the samples. Overall, 40/474 samples (8.44%) showed B1 gene positivity. T. gondii infection among the wild small mammals trapped in the rural area was significantly higher as a whole than that of the urban area as a whole. Multivariate logistical regression analysis also showed that the trapping area (rural or urban) significantly contributed to T. gondii positivity. Vegetation in the trapping points, small mammal species, sex, age or distance from the trapping points to the nearest human settlements did not significantly affect T. gondii positivity in the sampled small mammals.

Targeting the B1 Gene and Analysis of Its Polymorphism Associated with Awned/Awnless Trait in Russian Germplasm Collections of Common Wheat

The presence of awns on the ear is associated with a number of important plant properties, such as drought resistance, quality of the grain mass during processing, etc. The main manifestations of this trait are controlled by the B1 gene, which has recently been identified and encodes the C2H2 zinc finger transcription factor. Based on the previously identified SNPs in the promoter region of this gene, we constructed markers for dominant and recessive alleles which determine awnless and awned phenotypes, respectively. The markers were successful for use in targeting the respective alleles of the B1 gene in 176 varieties of common wheat, accessions of T. spelta L., as well as on F2/F3 hybrids from crosses between awned and awnless forms of T. aestivum. We first identified a new allele, b1mite, which has both an insert of a miniature Stowaway-like transposon, 261 bp in length, and 33 novel SNPs in the promoter region. Despite these changes, this allele had no effect on the awned phenotype. The possible mechanisms of the influence of the analyzed gene on phenotype are discussed.

Genetic Diversity of Toxoplasma Gondii by Serological and Molecular Analyzes in Different Sheep and Goat Tissues in Northeastern Iran

Background: Toxoplasmosis, a parasitic disease caused by compilation protozoan agent Toxoplasma gondiithat led to significant financial and quality-adjusted life-year losses. Consumption of undercooked or raw meat has been regarded as a major route of transmission. The present study was conducted to determine the seroposevitity rate of T.gondii in sheep and goats by serological and molecular tests and also genotyping of obtained isolatesin northeast of Iran.Methods: Blood and tissue samples (diaphragm, heart) of 296 animals (including 168 sheep and 128 goats) were collected from slaughterhouse in Quechan city from august 2016 to April 2017. Serum samples examined by the Modified agglutination test (MAT) and the Nested-PCR method performed to amplify the fragment of the B1 gene to detect parasite DNA on diaphragm and heart tissues of seropositive animals. PCR-RFLP method of GRA6 gene was used to determine the genotype of T. gondii. Also, sequencing analysis was performed to evaluate the Toxoplasma type strains. Results: Serum positive for MAT results were found in 27.4% (46/168) of Sheep and23.4% (30/128) of goats. Positive Nested-PCR of B1 gene results in diaphragm and heart tissues of sheep and goats was 47.8% (22/46) and 26.1% (12/46), 40% (12/30) and 23.3% (7/30), respectively. Nested-PCR of GRA6 gene results were positive in 10 samples (7 sheep and 3 goats) that RFLP technique results with using MseІ enzyme revealed genotype І. Sequencing and Phylogenetic analysis revealed DNA of all samples were closely related to Toxoplasma type І.Conclusions: Concerning to highseropositivityrate of toxoplasmosis in studied region, undertaking an appropriate preventive program for reducing the prevalence of T. gondii infection by raw or undercooked meat consumption of livestock recommended. Our study supports the notion that consumption of raw and undercooked meat of these animals can be a probable source of human toxoplasmosis.

The Response of Wheat with Different Allele Statuses of the Gpc-B1 Gene under Zinc Deficiency

The aim of this study was to investigate the effect of zinc (Zn) deficiency on the growth and grain yield of wheat with different allele statuses of the Gpc-B1 gene. For this research, common wild emmer wheat (Triticum turgidum ssp. dicoccoides (Koern. ex Asch. &Graebn.) Schweinf.), bread wheat (Triticum aestivum L. cv. Festivalnaya), and two intogressive lines were used. T. dicoccoides and introgressive line 15-7-1 carry a functional allele of the Gpc-B1 gene, while the T. aestivum cv. Festivalnaya and introgressive line 15-7-2 carry the non-functional Gpc-B1 allele. Zn deficiency did not affect the shoot height or fresh weight of any of the studied plants. The only exception was T. dicoccoides, where a small decrease in shoot height was registered. Additionally, under Zn deficiency T. dicoccoides had an increase in flag leaf area, spike length, and dry weight, as well as in grain number and grain yield per spike. The other variants did not experience changes in the above-described parameters under Zn deficiency. Under Zn deficiency, the Zn concentration in the grains was higher in the plants with a functional allele of the Gpc-B1 gene compared to the plants with a non-functional allele. These results show that wheat with a functional allele of the Gpc-B1 gene growing under Zn deficiency is capable of grain production with a sufficient Zn concentration without a decrease in yield.

Molecular Detection of Toxoplasma gondii and Neospora caninum in Domestic Ducks in Hunan Province, China

Toxoplasma gondii and Neospora caninum are protozoan parasites that infect warm-blooded animals, and cause major economic losses in livestock industries worldwide. However, little is known about the genotypes of T. gondii and N. caninum in domestic ducks in China. Herein, brain samples from 588 domestic ducks from Hunan province in China were examined for the presence of T. gondii and N. caninum. Polymerase chain reaction (PCR) was used to detect T. gondii B1 gene and N. caninum NC-5 gene. Forty-five DNA samples (7.7%; 95% CI: 5.5–9.9) were positive for B1 gene, and two (0.3%; 95% CI: 0–0.7) were positive for NC-5 gene. The risk factors significantly associated with T. gondii infection were age and sex. The 45 samples positive for T. gondii were genotyped using multi-locus PCR-RFLP analysis and only one sample was fully genotyped as ToxoDB#9 (Chinese I). These results provide new information about the epidemiology of T. gondii and N. caninum in ducks in Hunan province in China. The data also highlight the importance of a “One Health” approach to dealing with toxoplasmosis.