Effect of dipotassium-ethylenediaminetetraacetic acid or lithium-heparin treatments and storage times on selected clinicopathologic analytes in equine synovial fluid.

BACKGROUND: Sample processing methods and storage time affect the outcome of biochemical analysis. OBJECTIVES: We aimed to assess the effects of dipotassium-ethylenediaminetetraacetic acid (K2-EDTA) and lithium-heparin treatments and storage times on selected analytes in equine synovial fluid (SF). METHODS: Approximately 2 mL of SF from each horse (n = 7) were collected via femoropatellar joint arthrocentesis into K2-EDTA-treated bottles (K2-EDTA group), lithium-heparin-treated bottles (heparin group), and plain bottles (control group). The pH was determined using an electronic bench pH meter. The total nucleated cell count (TNCC) of samples was determined by hemocytometer method, while total protein (TP) concentrations, and lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) activities of the samples were determined spectrophotometrically at 2, 8, 24, 48, and 168 hours postcollection while being maintained at approximately 4°C. RESULTS: TP concentrations in the anticoagulant-treated groups remained stable for 48 hours. TNCCs were stable for 8 hours. However, after 2 hours, ALP, LDH, and pH varied significantly (P < 0.05). At 2 hours, mean ALP and LDH activities were significantly elevated in the lithium-heparin treatment samples, while the activity of these analytes was similar in the K2-EDTA and control groups. At 8 hours, the TNCC and pH were significantly elevated in K2-EDTA treated groups, while values were similar in lithium-heparin and control groups. No significant variation was seen in TP values at 2 hours, irrespective of treatment. CONCLUSIONS: The analytes-except for TP-became unstable within a few hours postcollection. Lithium-heparin and K2-EDTA treatments significantly altered ALP, LDH, TNCCs, and pH but not the TP concentrations of equine SF. Studies establishing reference intervals for these analytes based on the anticoagulant used are warranted to limit misinterpretations in clinical or research settings.

Two New Antiprotozoal Diterpenes From the Roots of Acacia nilotica.

The powdered roots of the medicinal plant Acacia nilotica were extracted with hexane and ethyl acetate, and the extracts were subjected to column chromatography for the isolation of potentially bioactive compounds and their screening against kinetoplastid pathogens. NMR and HREI mass spectrometric analyses identified two new diterpenes, characterized as 16, 19-dihydroxycassa-12-en-15-one (Sandynone, 1) and (5S, 7R, 8R, 9R, 10S, 13Z, 17S)-7,8:7,17:16,17-triepoxy-7,8-seco-cassa-13-ene (niloticane B, 2). The previously reported (5S,7R,8R,9R,10S) -(-)-7,8-seco-7, 8-oxacassa-13,15-diene-7,17-diol (3), (5S,7R,8R,9R,10S) -(-)-7,8-seco-7, 8-oxacassa-13,15-dien-7-ol-17-al (4), and (5S,7R,8R,9R,10S) -(-)-7,8-seco-7, 8-oxacassa-13,15-dien-7-ol (5) a, mixture of stigmasterol (6a) and sitosterol (6b), and lupeol (7) were also isolated. Several column fractions displayed significant activity against a panel of Trypanosoma and Leishmania spp., and from the most active fraction, compound 4 was isolated with high purity. The compound displayed high activity, particularly against T. brucei, T. evansi, and L. mexicana (0.88-11.7 µM) but only a modest effect against human embryonic kidney cells and no cross-resistance with the commonly used melaminophenyl arsenical and diamidine classes of trypanocides. The effect of compound 4 against L. mexicana promastigotes was irreversible after a 5-h exposure, leading to the sterilization of the culture between 24 and 48 h.

Anthelmintic potential of three plants used in Nigerian ethnoveterinary medicine.

CONTEXT: The leaves of Irvingia gabonensis Baill. Ex Lanen (Irvingiaceae), Ficus exasperata Vahl (Moraceae), and Vernonia amygdalina Delile (Asteraceae) are folklorically used in treating worm infestation in Eastern Nigeria. The anthelmintic potential of the ethanol extracts of the leaves of I. gabonensis, F. exasperata, and V. amygdalina was investigated. MATERIALS: Acute toxicity tests were done in mice using 10, 100, and 1000 mg/kg/bw of extracts. In vitro larval assays of Heligmosomoides bakeri larvae at various extract concentrations (125, 250, and 500 mg/kg) were done. Mice experimentally infected with H. bakeri were treated with F. exasperata extract (200, 400, 800 mg/kg). RESULTS: At concentrations of 500, 250, and 125 mg/ml F. exasperata caused 100% larval mortality. V. amygdalina extract caused 71.43, 57.14, and 57.14% larval deaths while I. gabonensis extract caused 71.43, 57.14, and 42.9% larval deaths at the same concentrations. There was no significant difference in the fecal egg output, packed cell volumes and body weights of the F. exasperata treated mice when compared with the infected untreated group. DISCUSSION AND CONCLUSION: Leaf extracts of F. exasperata, V. amygdalina, and I. gabonensis exhibited varying degrees of larvicidal activities on the infective stage larvae of H. bakeri in vitro whereas F. exasperata showed no activity on the parasites in vivo.