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The effects of human chorionic gonadotrophin (hCG) and a combination of growth factors on the developmental competence of canine oocytes during in vitro maturation was examined. Oocytes recovered from domestic dog ovaries at routine ovariectomy were cultured in a basic tissue culture medium with 0.3% BSA, 7 microg mL(-1) progesterone and antibiotics. After the appropriate culture periods (up to 96 h), they were fixed and labelled by double-antibody immunofluorescence for tubulin and with propidium iodide for chromatin. Human chorionic gonadotrophin increased the proportion of oocytes resuming meiosis and reduced the degeneration rate. Supplementing with hCG in declining concentrations was of no superior benefit but the presence of a combination of growth factors (growth hormone, insulin-like growth factor-1, transforming growth factor-alpha and fibroblast growth factor) improved both the resumption of meiosis and the degeneration rate. No particular synergisms between pairs of growth factors could be demonstrated. Human chorionic gonadotrophin and growth factors together gave poorer results, implying that hCG inhibited the beneficial effects of the growth factors. A growth factor combination is the present most successful treatment, with 49% of total oocytes (inclusive of degenerated) recovered from anoestrous bitches at MI or MII by 96 h of culture. This is the highest result so far demonstrated for cultured dog oocytes.

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Cryptosporidium parvum (C. parvum) infection is one of the main causes of diarrhea in calves. The current study assessed the role of blood biomarkers (acute-phase proteins (APPs), procalcitonin, neopterin, cytokines, and oxidative stress in the pathogenesis, diagnosis, and prognosis of calves naturally infected with C. parvum. Fifty-seven calves, aged from 10 to 45 days, were detected positive for C. parvum and were allotted into the diseased group; twenty healthy calves were selected as a control group. Serum amyloid A, haptoglobin, cytokines, neopterin, procalcitonin, and stress biomarkers were tested in the diseased and healthy groups. The serum levels of APPs, cytokines, neopterin, procalcitonin, and malondialdehyde increased, whereas antioxidant levels were significantly decreased in diseased calves compared to the healthy group. Moreover, all examined biomarkers were significantly altered towards normal values in infected calves following different treatment protocols. All biomarkers examined were highly effective in discriminating between C. parvum-infected calves and healthy individuals. Furthermore, the area under the curve (AUC) showed that all tested parameters had a higher degree of prognostic accuracy in predicting the treatment response of calves naturally infected with C. parvum. Our data suggest the usefulness of the examined biomarkers in the immune pathogenesis of the C. parvum infection in calves, contributing to diagnosis and treatment efficacy.