The severity of the clinical manifestations of cutaneous leishmaniasis can vary depending on various factors, such as the Leishmania species involved as well as hosts and their immune response. This study aimed to investigate the relationship between the severity of different clinical signs, histopathological changes, and genetic indicators with TAC in mice experimentally infected with Leishmania major.A total of 105 eight-week-old BALB/c mice of both sexes were assigned to seven experimental groups (15 in each) as follows: 1) healthy mice, 2) Leishmania-infected mice treated with 100 mg/kg/day of SC glucantime until complete healing, 3) mice which received 20 IU/kg/day of vitamin E (SC for 10 days) to increase TAC prior to infection and further treatment with glucantime, 4) Leishmania-infected mice which received both vitamin E and glucantime daily until complete healing, 5) mice which received 20 IU/kg/day of vitamin E (SC for 10 days) before infection, and 6) Leishmania-infected mice which received 20 IU/kg/day of SC vitamin E up to the end of the trial, and 7) mice which received daily vitamin E until the end of the experiment. Wound size, expression of pro-inflammatory cytokines (IFN-γ and TNF-α) and healing genes (KGF and EGF), histopathological findings, and mortality rate were assessed three times on days 31, 38, and 72 post-infection. Approximately, 31 days after the parasite inoculation, dermal lesions were developed in all infected mice. In group 3, the clinical manifestations, healing time, and histopathological changes were significantly more favorable, while group 4 showed the worst situation in terms of the evaluated indicators. A high level of TAC before the onset of the disease has an effective role in the recovery indicators. However, its simultaneous elevation at the beginning of infection will decrease the body's ability to effectively clear the parasite, heal the tissue, and improve the clinical manifestations of the disease.

Paratuberculosis (Johne's disease) is a chronic granulomatous small intestine disease caused by MAP. Diagnosing and isolating infected animals is the most important measure for controlling the disease. Therefore, this study aimed to molecularly identify mycobacterium isolated from ELISA-positive cows with Johne's disease by nested PCR from the samples from Markazi Province, Iran. For this purpose, 2938 samples were decontaminated and then cultured on the Herrold egg culture medium containing mycobactin and no mycobactin. After DNA extraction, PCR for 16S rRNA was first performed, followed by nested PCR on positive samples. Of 2938 samples, 87 were positive, and 26 were suspected. All positive isolates were observed in Ziehl-Neelsen staining in microscopic expansion. A 543-bp band was observed in 26 tested samples and mycobacterium strains in PCR for 16S rRNA, indicating the presence of mycobacterium in the above samples. Nested PCR was performed for all isolates and positive and negative control strains. A 398-bp band was obtained in the first stage, and a 298-bp fragment was obtained in the second stage, indicating the presence of MAP in the samples. Accordingly, nested PCR is suggested as a proper method for the quick and definitive diagnosis of disease cases.