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Results for “Analytical Biochemistry”

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USGS1993

Polyphosphate present in DNA preparations from fungal species of Collectotrichum inhibits restriction endonucleases and other enzymes

During the development of a procedure for the isolation of total genomic DNA from filamentous fungi (Rodriguez, R. J., and Yoder, 0. C., Exp. Mycol. 15, 232-242, 1991) a cell fraction was isolated which inhibited the digestion of DNA by restriction enzymes. After elimination of DNA, RNA, proteins, and lipids, the active compound was purified by gel filtration to yield a single fraction capable of complete inhibition of restriction enzyme activity. The inhibitor did not absorb uv light above 220 nm, and was resistant to alkali and acid at 25°C and to temperatures as high as 100°C. More extensive analyses demonstrated that the inhibitor was also capable of inhibiting T4 DNA ligase and Taq I DNA polymerase, but not DNase or RNase. Chemical analyses indicated that the inhibitor was devoid of carbohydrates, proteins, lipids, and nucleic acids but rich in phosphorus. A combination of nuclear magnetic resonance, metachromatic shift of toluidine blue, and gel filtration indicated that the inhibitor was a polyphosphate (polyP) containing approximately 60 phosphate molecules. The mechanism of inhibition appeared to involve complexing of polyP to the enzymatic proteins. All species of Colletotrichum analyzed produced polyP equivalent in chain length and concentration. A modification to the original DNA extraction procedure is described which eliminates polyP and reduces the time necessary to obtain DNA of sufficient purity for restriction enzyme digestion and Taq I polymerase amplification.

Analytical Biochemistry↗

USGS2022

Blood biochemistry and hematology of adult and chick brown pelicans in the northern Gulf of Mexico: Baseline health values and ecological relationships

The northern Gulf of Mexico supports a diverse community of nearshore seabirds during both breeding and nonbreeding periods of the annual cycle and is also a highly industrialized marine ecosystem with substantial levels of oil and gas development particularly in the west and central regions. Stakeholders in the region often assess risk to species of interest based on these differing levels of development. We collected blood samples from 81 adult and 35 chick eastern brown pelicans ( Pelecanus occidentalis carolinensis ) from 10 colonies across the northern Gulf of Mexico and used these to establish baseline values for hematology and blood biochemistry. We assessed the potential influence of body condition, sex and home range size on hematology and blood biochemistry. We also assessed potential influences of oil and gas activity by considering differing levels of oil and gas development that occur regionally throughout the study area. Although blood analyte concentrations of adults and chicks were often associated with these regional differences, the pattern we observed was not entirely consistent with the differing levels of oil and gas activity across the Gulf, suggesting that regional levels of oil and gas activity around breeding sites may not be the primary drivers of hematology and blood biochemistry. We note that baseline values or reference intervals are not available for other nearshore seabirds that breed in the northern Gulf. Given that exposure and risk may differ among this suite of species based on diet, foraging strategies and life history strategies, similar assessments and monitoring may be warranted.

Alabama, Florida, Louisiana, Mississippi, Texas↗

USGS2007

Clinical biochemistry in healthy manatees (Trichechus manatus latirostris)

Florida manatees (Trichechus manatus latirostris) are endangered aquatic mammals living in coastal and riverine waterways of Florida and adjacent states. Serum or plasma biochemical analyses are important tools in evaluating the health of free-ranging and captive manatees. The purpose of this study was to measure diagnostically important analytes in the plasma of healthy manatees and to determine whether there was significant variation with respect to location (free-ranging versus captive), age class (small calves, large calves, subadults, adults), and gender. No significant differences in plasma sodium, potassium, bilirubin, glucose, alanine aminotransferase, or creatine kinase were found among these classes of animals. Compared to free-ranging manatees, captive animals had significantly lower mean concentrations of plasma chloride, phosphate, magnesium, triglycerides, anion gap, and lactate. Captive manatees had significantly higher mean values of total CO2, calcium, urea, creatinine, alkaline phosphatase, gamma-glutamyltransferase, total protein, albumin, and albumin/globulin ratio than did free-ranging animals. Differences in the environments of these two groups, including diet, temperature, salinity, and stress, might account for some of these results. The higher plasma lactate and anion gap concentrations and lower total CO2 concentrations of free-ranging manatees were probably due to greater exertion during capture, but the lack of elevated plasma creatine kinase activity relative to captive animals indicates that there was no serious muscle injury associated with capture. Plasma phosphate decreased and total globulins increased with age. Plasma cholesterol and triglyceride concentrations were highest in small calves. Plasma aspartate aminotransferase was higher in large calves than in adults and subadults, and the albumin/ globulin ratio was higher in subadults than in adults. Plasma total CO2 was higher and chloride was slightly lower in females than in males.

Journal of Zoo and Wildlife Medicine↗