Figure 6. Disease score for visceral leishmaniasis.

A score to measure the grade of severity of the disease was performed for 3%INF (A), 14%INF (B), 3%NI (C) and 14%NI (D). For each hamster, a score of 2 was given to values below normal for leukocytes, lymphocytes, PLT and albumin; and to values above normal for total proteins, BUN, AST, globulin and erythrocytes. Other abnormalities in these biochemical (BioCh) or blood variables were scored as 1. Lymphoproliferative responses for SLA and CONA were scored as: 3 for a stimulation index (SI) <0.60; 2 for 0.60≤ SI<2.00; 1 for 2.00≤ SI<4.00; and 0 for SI≥4.00. The humoral response as determined by ELISA was scored as: 1 for 0.10≤ OD 430 nm <0.60; 2 for 0.60≤ OD 430 nm <1.00; 3 for OD 430 nm≥1.00. As determined by IFAT, the humoral response was scored as: 1 for 1/80 titre; 2 for 1/160 titre; 3 for 1/320 titre. Spleen and bone marrow parasite loads as determined by qPCR were scored as: 1 for 35≤ parasites/μg/ml<1000; 2 for 1000≤ parasites/μg/ml<5000; 3 for parasites/μg/ml≥5000. Liver and lymph node parasite loads were scored as: 1 for 35≤ parasites/μg/ml<600; 2 for parasites/μg/ml≥600. Presence of Leishmania DNA in the blood was given a disease score of 2. Histopathology for liver and spleen was scored from 0 to 3. A corrective factor was applied to the 6 categories of data to compensate the global score. Visceral leishmaniasis status was recorded as: Mild, when the total score for an animal was 2–2.5; Moderate, when between 2.6–3.0; and Severe, when >3.1. The bars represented each hamster and showed the contribution of each parameter to the total score. Abbreviations: BioCh- serum biochemistry profile, Blood- haematological alterations, Cell Rsp- lymphoproliferative response to SLA and CONA, Hum Rsp- ELISA and IFAT results, qPCR- parasite load (blood, lymph node, spleen, liver and bone marrow), and Histology- grade of alterations in spleen and liver.