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Figure 2 | Veterinary Research

Figure 2

From: Extensive production of Neospora caninum tissue cysts in a carnivorous marsupial succumbing to experimental neosporosis

Figure 2

Neosporosis in fat-tailed dunnarts. Appearance of the infected dunnarts (a). The fur of infected dunnarts was roughened due to marked incontinence (A), recognised as wet anal, perineal and base of the tail regions (B22). The negative control animal (B2C) had clean groomed fur including around the tail base (B). Interscapular (arrow) and gluteal (arrowheads) fat was absent from infected animals (B21, B22, B23) compared to the control animal (B2C) which had marked fat deposition at both sites (C). PCR detected presence of N. caninum DNA using Np6+/Np21+ primers (b). A N. caninum specific reaction appeared as a 330 bp amplicon. In experiment A the infectious dose was 104 N. caninum tachyzoites (1-3); this experiment included a N. caninum-infected animal immunosuppressed with methylprednisolone acetate as a control (S) plus an animal not infected with N. caninum as a negative control (C). Animals in experiments B1 and B2 were inoculated with 105 N. caninum tachyzoites (1-3). Animals in experiment E were inoculated orally with less than 20-40 oocysts of N. caninum (1, 2). Negative (-, water) and positive (+, N. caninum DNA) controls were included for each PCR batch. The left lane is a 100 bp DNA ladder (300, 400, 500 bp). Gel is stained using GelRed and inspected under UV. Oil Red O staining of tails of fat-tailed dunnarts (c) and ImageJ colour threshold applied to separate red areas of fat from the skin, muscle and bone for statistical analysis (d). Tail cross sections of animals in experiments B1 and B2 (c). Ratio means were compared for different inoculation doses (104 N. caninum tachyzoites, n = 3; 105 N. caninum tachyzoites, n = 6; < 50 N. caninum oocysts, n = 2; control animals, n = 3) and comparison made using Student's t-Test (P < 0.05 is considered significant).

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