Simulation of photodynamically-induced thrombosis of the femoral vein of rats

The purpose of the work is to determine the conditions that ensure the formation of a stable thrombus in the femoral vein of rats with photodynamic damage.Material and methods. The experiments were carried out on male Wistar rats weighing 250–270 g. A photosensitizer (FS) Bengal rose (ACROS ORGANICS, USA) (BR) was administered intravenously at a dose of 17 mg/kg and after 60 minutes a vascular clamp was applied to the femoral vein. Distal to clamping, a 30-minute laser irradiation of a vein was performed (DPSS diode laser, Republic of South Korea, 532 nm, 60 mW, irradiation area – 1 mm2 ). The clamp was removed after 90 min after cessation of irradiation. The blood flow was recorded by the method of high-frequency Doppler ultrasound (MinimaxDoppler-K Russia). Cessation of blood flow was considered as an indicator of occlusion of femoral vein.Results. Rats, after the clamp was removed, had no blood flow in the irradiated veins, and after 24 hours it was not restored. Histology revealed severe damage of all layers of the vascular wall and the formation of a blood clot filling the lumen of the vessel. After a 30-minute irradiation without clamping the vein, the blood flow was maintained, although damage to the vascular wall was detected. Introduction of BR and clipping of the vessel without irradiation caused minimal damage to the vascular wall and did not lead to thrombosis.

Sperm morphological and morphometric evaluation in captive collared peccaries (Pecari tajacu)

The aim of this study was to compare different staining methods for the evaluation of sperm morphology by light microscopy and also to describe the morphometry of the entire sperm in collared peccaries (Pecari tajacu). Semen from 10 males was obtained by electroejaculation and evaluated for sperm motility, vigor, and concentration. Semen smears were prepared through three different staining methods: Bengal rose, brome-phenol blue, and eosin-nigrosin. Smears were evaluated under light microscopy and sperm morphologic alterations were determined in percentage. In addition, sperm morphometric analysis was conducted by light microscopy coupled to image analyzer software. The smears stained with Bengal Rose provide the best results for the visualization of the sperm tail, midpiece, and head. The use of eosin-nigrosin stain did not allow an adequate impregnation, and some sperm presented a few contrasts with the background. A higher incidence of bent coiled tails was verified in the use of brome-phenol blue staining (P<0.05). Through morphometric evaluation, it was observed that the tail occupies the greatest proportion (89%) of the sperm which presents a discretely elongated head. According to the results, the use of the Bengal Rose stain is recommended for the morphologic evaluation of the collared peccary sperm.