Brain-derived neurotrophic factor (BDNF) can promote developmental competence in mammalian oocytes during in vitro maturation (IVM), but the role of BDNF in oocyte maturation at cellular level is not still clear. In this study, mouse cumulus-enclosed oocytes subjected to IVM were fertilized and cultured to blastocyst stage. Meiotic spindle configuration and cortical granules distribution during oocyte maturation in vitro were assessed by using immunofluorescence and laser confocal microscopy. The results showed that BDNF contributed to the complete preimplantation development of mouse oocytes compared to the control oocytes (13.78% vs. 5.92%; P<0.05). Further, BDNF did not accelerate nuclear maturation of IVM oocytes. For the BDNF-treated oocytes at meiosis I, Meiotic spindle areas were significantly smaller and the number of cytoplasmic microtubule organizing centers was greater than that in the control, and the percentages of oocytes showed spindles positioned near the oolemma and a well-formed cortical granule-free domain were significantly higher than that of the control. These morphological characteristics of the BDNF-treated oocytes were much closer to the oocytes matured in vivo than those of the control oocytes. In conclusion, BDNF can promote the developmental competence of mouse IVM oocytes, by improving the meiotic spindle configuration and location and cortical granules distribution at meiosis 1.
Animals ; Blastomeres ; cytology ; Brain-Derived Neurotrophic Factor ; pharmacology ; Female ; Fertilization in Vitro ; In Vitro Oocyte Maturation Techniques ; methods ; Male ; Mice ; Oocysts ; growth & development

Neospora caninum is an important cause of abortion in dairy cattle worldwide. Dog is the definitive host for N. caninum and can infect dairy cattle. The aim of this study is to determine the prevalence of Neospora oocysts in feces of dogs from dairy farms. A total of 174 fecal samples was collected from 89 farm dogs and 85 household dogs during 2006 and 2008. Fecal samples of dogs were microscopically examined for detecting Hammondia Neospora-like oocysts (HNLO) by Mini Parasep(R)SF fecal parasite concentrator. HNLO were microscopically detected in 4 fecal samples (2.2%). The fecal samples with HNLO were examined by N. caninum-specific PCR. Two of the samples were positive for N. caninum. The 2 positive fecal samples were selected for inoculation to calves. Two inoculated calves were seronegative by ELISA for 4 months post-infection. This is the first report of finding N. caninum DNA in feces of farm dogs in Mashhad area, Iran.
Animals ; Antibodies, Fungal/blood ; Cattle ; Cattle Diseases/immunology/parasitology ; Coccidiosis/epidemiology/parasitology/*veterinary ; DNA, Fungal/genetics/isolation & purification ; Dog Diseases/*epidemiology/*parasitology ; Dogs ; Enzyme-Linked Immunosorbent Assay/methods ; Feces/*microbiology ; Iran/epidemiology ; Male ; Microscopy/methods ; Neospora/*genetics/*isolation & purification ; Oocysts/cytology ; Polymerase Chain Reaction/methods ; Prevalence

The present study surveyed the prevalence of natural infection of the sheep esphagus muscle with sarcocysts of Sarcocystis ovicanis and examined induction of protective immunity using UV-attenuated sporocysts. The overall prevalence of natural infection of the sheep was 95%. Infectivity of the collected sarcocysts was confirmed by shedding of sporulated oocysts after feeding infected esophageal tissues to dogs. To induce protective immunity, lambs were immunized 3 times (once a week) with 1.5 x 10(4) sporocysts exposed to UV-light for 30 min (UV-30 group) or 60 (UV-60 group) min and then challenged with 1.5 x 10(4) normal sporocysts at the 3rd week post the 1st vaccination. These lambs showed high survival and less clinical signs of sarcocystosis than normal infected lambs. The attenuated sporocysts produced abnormal cysts; small in size and detached from the muscle fiber. These abnormalities were more obvious in UV-60 group than UV-30 group. Also, the IFN-gamma level and lymphocyte percentage were increased while the total leukocyte count was decreased in the UV-60 group compared with other groups. The high level of IFN-gamma may be an evidence for the induction of Th1 responses which may have protective effect against a challenge infection.
Animals ; Dogs ; Esophagus/parasitology ; Feces/parasitology ; Interferon-gamma/secretion ; Lymphocytes/immunology ; Oocysts/*immunology ; Peptide Fragments/secretion ; Prevalence ; Protozoan Vaccines/immunology ; Sarcocystis/cytology/*immunology/*radiation effects ; Sarcocystosis/epidemiology/immunology/prevention & control/*veterinary ; Severity of Illness Index ; Sheep/immunology/parasitology ; Sheep Diseases/immunology/*prevention & control ; Survival Analysis ; *Ultraviolet Rays ; Vaccines, Attenuated/immunology