Direct evidence of cheetah (Acinonyx jubatus) as intermediate host of Toxoplasma gondii through isolation of viable strains.

Toxoplasma gondii causes lifelong infection in most definitive and intermediate hosts. Clinical cases of toxoplasmosis in captive cheetahs have been reported. However, there are few reports of viable T. gondii strains isolated from cheetahs. Here, T. gondii infection was investigated using molecular and serological assays in cheetahs from China. Modified agglutination test (MAT) (cut-off: 1:25) indicated that all six examined cheetahs (n = 6) showed T. gondii antibodies. Toxoplasma gondii DNA was detected in three out of five cheetahs. Two viable T. gondii strains were isolated from the striated muscles of two cheetahs using mice bioassay. They were designated as TgCheetahCHn1 and TgCheetahCHn2. Genetic characterization of DNA derived from tachyzoites was performed using RFLP-PCR of 10 markers. Toxoplasma gondii TgCheetahCHn1 is ToxoDB PCR-RFLP genotype #319, and the alleles of ROP18/ROP5 types were 3/7. TgCheetahCHn2 is ToxoDB genotype #9, and the alleles of ROP18/ROP5 were 3/6. The average survival time of TgCheetahCHn1-infected Swiss mice was 22 ± 1 days (n = 23), and the mice did not have detectable T. gondii-specific antibodies until 117 ± 30 days post-inoculation (n = 8), therefore, TgCheetahCHn1 had intermediate virulence. TgCheetahCHn2 was avirulent for Swiss mice. Few brain tissue cysts (0-50) were observed in the mice inoculated with TgCheetahCHn1 or TgCheetahCHn2. The results provide direct evidence of cheetah as intermediate host of T. gondii.

Isolation and Characterization of a Viable Toxoplasma gondii from Captive Caracal (Caracal caracal).

Toxoplasma gondii is a widespread protozoan parasite that infects most warm-blooded animals, and felids can serve as intermediate and definitive hosts. Pathological diagnosis and serological and etiological investigations were conducted on a captive caracal (Caracal caracal) carcass collected from China in 2022. Pathological diagnosis revealed that cardiac insufficiency, pulmonary edema, hepatic failure, and renal insufficiency were the causes of the caracal's death. A modified agglutination test (cut-off: 1:25) revealed that IgG antibodies against T. gondii were present in the myocardium juice (1:1600), ascitic fluid (1:3200), and hydropericardium (1:800). A viable T. gondii (TgCaracalCHn2) strain was isolated from the tissue samples (heart, brain, spleen, and skeletal muscle) of this caracal using a mouse bioassay. The genotype of TgCaracalCHn2 was ToxoDB#5 (Type II variant), as determined via RFLP-PCR. The strain was avirulent in Swiss mice and matched the prediction of ROP18 and ROP5 gene alleles of TgCaracalCHn2 (2/2). Mild tissue cysts (203 ± 265) were observed in mice brains after inoculation with TgCaracalCHn2 tachyzoites. ToxoDB#5 is the dominant genotype in North American wildlife, and this is the first documented isolation of T. gondii ToxoDB#5 from China. This indicates that caracal plays an important role in the transmission of this T. gondii genotype.

Additional evidence of tigers (Panthera tigris altaica) as intermediate hosts for Toxoplasma gondii through the isolation of viable strains.

Toxoplasmosis is one of the most common zoonotic diseases in the world. Felines excrete Toxoplasma gondii oocysts, which play a key role in the transmission of this protozoon. Pathological diagnoses were performed on four carcasses of captive tigers collected from 2019 to 2021 in China, and T. gondii was surveyed using serology, molecular analysis, and aetiology. Striated muscle samples of the tigers (n = 4) were bioassayed in mice. DNA derived from T. gondii tachyzoites was isolated and characterized using PCR-RFLP. The pathological diagnoses revealed that ageing, declined immune function, liver, and kidney failures caused the deaths in the tigers examined. A modified agglutination test (cut-off: 1:25) revealed that IgG antibodies to T. gondii were 100% (4/4) in the captive tigers. Two viable T. gondii strains (TgTigerCHn3 and TgTigerCHn4) were isolated from tiger striated muscles and seeded on the Vero cell culture for further propagation. The genotypes of TgTigerCHn3 and TgTigerCHn4 were ToxoDB#20 and ToxoDB#2, respectively. The two strains were avirulent for Swiss mice, which matched the ROP18 and ROP5 gene alleles of TgtigerCHn3 (3/4) and TgtigerCHn4 (3/3). Few brain tissue cysts (0-213) were observed in the mice after inoculation with TgTigerCHn3 and TgTigerCHn4. This is the first documented isolation of T. gondii ToxoDB#20 and ToxoDB#2 from tigers. The results provide additional direct evidence of tiger as intermediate hosts for T. gondii. Tigers in the zoos may potentially transmit T. gondii to other animals and humans.

Isolation, genotyping and virulence determination of a Toxoplasma gondii strain from non-human primate from China.

Toxoplasma gondii infects almost all warm-blooded animals, including humans and non-human primates. Many cases of T. gondii infection in non-human primates have been reported worldwide. In this study, 15 monkeys were collected from zoos in Henan Province between 2016 and 2019. A modified agglutination test (MAT) (cut-off: 1:8) showed that 46.7% (7/15) of the heart juices had T. gondii IgG antibody transformation. One viable T. gondii strain was successfully isolated from the myocardium of a rhesus monkey by bioassay in mice. This strain was designated as TgMonkeyCHn1. The DNA of T. gondii tachyzoites was obtained using cell cultures, and the genotype of this strain was determined by PCR-RFLP with 10 markers and the virulence genes ROP5 and ROP18. The genotype and ROP18/ROP5 (3/6) of TgMonkeyCHn1 did not match any known genotypes. In addition, the TgMonkeyCHn1 formed low number of tissue cysts and was non-lethal to mice. To our knowledge, this is the first T. gondii strain isolated from Old World monkeys. Rhesus monkey is a new host record for T. gondii.