Monitoring Mice Behavior
Monitoring mice behavior
Behavioral testing started at 10 weeks post-infection. Male and female mice of both groups were tested in separate trials. The behavior of the mice was videotaped and all videos were blindly analyzed using observer scoring software. Between each mouse testing, feces and urine was removed and arena was detergent-cleaned and dried.
Open- Field test
The Open-Field test originally described by Hall and ballechey (1932) was used with modifications. The test consisted of a 60x60x40 cm box with a black plastic arena floor, divided into 16 symmetrical squares (15x15 cm each) (Figure 1). The mouse is placed in a corner square, facing the walls, and videotaped for 10 min. Mouse activity was assessed by the following parameters: …show more content…
Mice were bled out completely before brains were removed and blood was available for further serologic examination (Dubey 2010). Brains were dissected within 90 seconds of death and brain imprint smear was done immediately and fixed by dipping twice in methanol to be examined later for T. gondii tissue-stages. Each brain was divided into two equal halves (one for the histopathological examination and one for the neurochemical analysis). Both were kept frozen at -80 °C.
Identification of Toxoplasma gondii parasitic stages in brain imprints
Giemsa-stained preparations were performed according to Garcia et al. (1991). Pre-fixed slides were stained by Giemsa diluted as 1:10 in buffered distilled water for 10 minutes, then washed in buffered distilled water and air-dried. Stained-slides were examined microscopically under X40 and X100 powers. Histopathological Examination
Histopathological examination of brain specimens was done in order to confirm the presence of T. gondii cysts in the brains of mice. Brain samples fixed in 10 % neutral buffered formalin were processed according to Culling (1974). The histological technique included slicing at 5 um, tissue processing and paraffin wax embedding, sectioning, and staining with Hematoxylin and Eosin (H & E). The stained tissue sections were microscopically-examined under X40 and X100 for the identification of Toxoplasma
Behavioral testing started at 10 weeks post-infection. Male and female mice of both groups were tested in separate trials. The behavior of the mice was videotaped and all videos were blindly analyzed using observer scoring software. Between each mouse testing, feces and urine was removed and arena was detergent-cleaned and dried.
Open- Field test
The Open-Field test originally described by Hall and ballechey (1932) was used with modifications. The test consisted of a 60x60x40 cm box with a black plastic arena floor, divided into 16 symmetrical squares (15x15 cm each) (Figure 1). The mouse is placed in a corner square, facing the walls, and videotaped for 10 min. Mouse activity was assessed by the following parameters: …show more content…
Mice were bled out completely before brains were removed and blood was available for further serologic examination (Dubey 2010). Brains were dissected within 90 seconds of death and brain imprint smear was done immediately and fixed by dipping twice in methanol to be examined later for T. gondii tissue-stages. Each brain was divided into two equal halves (one for the histopathological examination and one for the neurochemical analysis). Both were kept frozen at -80 °C.
Identification of Toxoplasma gondii parasitic stages in brain imprints
Giemsa-stained preparations were performed according to Garcia et al. (1991). Pre-fixed slides were stained by Giemsa diluted as 1:10 in buffered distilled water for 10 minutes, then washed in buffered distilled water and air-dried. Stained-slides were examined microscopically under X40 and X100 powers. Histopathological Examination
Histopathological examination of brain specimens was done in order to confirm the presence of T. gondii cysts in the brains of mice. Brain samples fixed in 10 % neutral buffered formalin were processed according to Culling (1974). The histological technique included slicing at 5 um, tissue processing and paraffin wax embedding, sectioning, and staining with Hematoxylin and Eosin (H & E). The stained tissue sections were microscopically-examined under X40 and X100 for the identification of Toxoplasma