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Forty-five cattle of different ages and gender were selected from three separate farms with a total number of 929 animals. Blood serum samples from each of the animals were tested twice at two-month intervals for bovine viral diarrhoea virus (BVDV) antigen (BVDV Ag) and BVDV antibodies (BVDV Ab) using ELISA. Five animals were found to be BVDV Ag positive and BVDV Ab negative. Therefore, their blood and saliva samples were subjected to further investigation. The samples of blood serum and saliva were additionally screened by a nested reverse transcription PCR (RT-nPCR), real-time PCR, and virus isolation to confirm BVDV persistent infection. Viral RNA was isolated from blood and saliva samples. The cDNA was synthesised and amplification of DNA was performed. The results of RT-nPCR were analysed by gel electrophoresis using ethidium bromide while those of real-time PCR were interpreted according to the amplification curve. Laboratory testing of blood and saliva samples revealed 5 persistently infected (PI) animals from one farm with 579 cattle (0.9% prevalence). The results were confirmed by RT-nPCR and real-time PCR screening samples of blood serum. Using PCR techniques and virus isolation, BVDV RNA was detected; however, the level of viral RNA in saliva was found to be lower than that in blood serum. The results obtained show the possibility to identify PI animals by RT-nPCR and real-time PCR techniques from saliva samples. The collection and testing of saliva is a simple and quick technique, and can be successfully applied in field conditions to identify PI animals, avoiding the risk of intervention while sampling blood or dependence on animal gender and lactation period while sampling milk or semen.
The aim of this study was to demonstrate the oceurrence of bovine foamy virus (BFV) and BFV DNA in peripheral blood leukocytes (PBLs), milk cells, and saliva of cattle serologically positive to BFV. The virus was detected by co-cultivation technique with canine thymus cells (Cf2Th) and viral DNA was quantified by the real-time PCR. Out of co-cultures from 23 cattle, BFV was found in 19, 13, and 15 samples of PBLs. milk cells and saliva, respectively, while BFV-DNA was confirmed in DNA extracted from 14 PBLs and 8 milk cells samples. All 23 saliva samples were negative. The number of BFV DNA copies in milk cells was in average over 2.5 times lower than in PBLs. The presence of BFV and viral DNA in milk and saliva samples is discussed in the context of virus spread and possible human exposure to BFV through food products of cattle origin.
Calcium and magnesium are known to be necessary for the normal function of various systems in animal and human organisms. There are many diseases caused by abnormal concentration of electrolytes, e.g. arterial hypertension or nervous system diseases such as multiple sclerosis, Mb. Alzheimer or Mb. Parkinson. The mechanisms of homeostasis indicate only the ionized forms of these elements. It is known that ionized calcium serves as an endocellular intermediary in action of enzymes and hormones in cells. Therefore, it is very important to define levels of total and ionized forms of Ca2+ and Mg2+ in blood serum and saliva by the method of atomic absorption spectrometry and to show their diagnostic value for various pathological conditions of a human body. The 39 persons, aged 21 to 47 years take part in these investigations. The results of determinations of calcium and magnesium forms present in human serum and saliva, representing physiological states are presented. The age and daily fluctuations of Ca2+ and Mg2+ content in serum and saliva were studied by atomic absorption spectrometry. The levels of non albumin forms of these elements were found by FAAS. The significance of determination of calcium and magnesium levels in serum and saliva under various pathological conditions (arterial hypertension and osteoporosis) was shown.
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