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Effect of anaesthetics MS-222 and clove oil on blood biochemical parameters of juvenile Siberian sturgeon (Acipenser baerii)

The effects of MS-222 and clove oil on blood biochemical parameters of juvenile Siberian sturgeon (Acipenser baerii) were studied. MS-222 caused higher glucose (GLU) concentrations in anaesthetic test groups than for the control group. Triglyceride (TGL) concentrations of fish in the 140 and 160mgL-1 groups were also significantly higher than those of other groups. Alanine aminotransferase (ALT) activity in the 140mgL-1 group was significantly higher than the level in 80, 100 and 120mgL-1 groups. Aspartate aminotransferase (AST) activity in the 140mgL-1 group was significantly higher than those in the 100 and 120mgL-1 groups. Levels of total protein (TP), cholesterol (CHOL) and alkaline phosphatase (ALP) in anaesthetic test groups were not significantly influenced by MS-222. Clove oil did not have significant effects on levels of GLU, TP, CHOL, ALT and ALP. TGL concentration of fish exposed to 180mgL-1 clove oil was significantly higher than those of the rest anaesthetic groups. AST activities of fish exposed to 120, 150 and 180mgL-1 were significantly higher than those of 60 and 90mgL-1. Overall, TGL and AST could be potentially used as indicators of anaesthetic stress for juvenile Siberian sturgeon. Based on blood biochemical parameters, the appropriate anaesthetic concentrations of MS-222 and clove oil were 80-120mgL-1 and 60-90mgL-1, respectively. Clove oil was a promising alternative to MS-222. ?? 2011 Blackwell Verlag, Berlin.

Journal of Applied Ichthyology

Statistical methods for estimating normal blood chemistry ranges and variance in rainbow trout (Salmo gairdneri), Shasta Strain

Gaussian and nonparametric (percentile estimate and tolerance interval) statistical methods were used to estimate normal ranges for blood chemistry (bicarbonate, bilirubin, calcium, hematocrit, hemoglobin, magnesium, mean cell hemoglobin concentration, osmolality, inorganic phosphorus, and pH for juvenile rainbow ( Salmo gairdneri , Shasta strain) trout held under defined environmental conditions. The percentile estimate and Gaussian methods gave similar normal ranges, whereas the tolerance interval method gave consistently wider ranges for all blood variables except hemoglobin. If the underlying frequency distribution is unknown, the percentile estimate procedure would be the method of choice.

Journal of the Fisheries Research Board of Canada

Sex identification of polar bears from blood and tissue samples

Polar bears ( Ursus maritimus ) can be adversely affected by hunting and other human perturbations because of low population densities and low reproduction rates. The sustainable take of adult females may be as low as 1.5% of the population. Females and accompanying young are most vulnerable to hunting, and hunters have not consistently reported the sex composition of the harvest, therefore a method to confirm the sexes of polar bears harvested in Alaska is needed. Evidence of the sex of harvested animals is often not available, but blood or other tissue samples often are. We extracted DNA from tissue and blood samples, and amplified segments of zinc finger (ZFX and ZFY) genes from both X and Y chromosomes with the polymerase chain reaction. Digestion of amplified portions of the X chromosome with the restriction enzyme Hae III resulted in subdivision of the original amplified segment into four smaller fragments. Digestion with Hae III did not subdivide the original segment amplified from the Y chromosome. The differing fragment sizes produced patterns in gel electrophoresis that distinguished samples from male and female bears 100% of the time. This technique is applicable to the investigation of many wildlife management and research questions.

Alaska

Prevalence and diversity of avian blood parasites in a resident northern passerine

Background: Climate-related changes are expected to influence the prevalence and distribution of vector-borne haemosporidian parasites at northern latitudes, although baseline information about resident birds is still lacking. In this study, we investigated prevalence and genetic diversity of Plasmodium, Haemoproteus, and Leucocytozoon parasites infecting the Northwestern Crow (Corvus caurinus), a non-migratory passerine with unique life-history characteristics. This species occupies both intertidal and forested habitats and is subject to high prevalence of avian keratin disorder (AKD), a disease that causes gross beak deformities. Investigation of avian blood parasites in Northwestern Crows at sites broadly distributed across coastal Alaska provided an opportunity to evaluate specific host factors related to parasite infection status and assess geographic patterns of prevalence. Results: We used molecular methods to screen for haemosporidian parasites in Northwestern Crows and estimated genus-specific parasite prevalence with occupancy modeling that accounts for imperfect detection of parasite infection. We observed considerable geographic and annual variation in prevalence of Plasmodium, Haemoproteus, and Leucocytozoon , but these patterns were not correlated with indices of local climatic conditions. Our models also did not provide support for relationships between the probability of parasite infection and body condition or the occurrence of co-infections with other parasite genera or clinical signs of AKD. In our phylogenetic analyses, we identified multiple lineages of each parasite genus, with Leucocytozoon showing greater diversity than Plasmodium or Haemoproteus. Conclusions: Results from this study expand our knowledge about the prevalence and diversity of avian blood parasites in northern resident birds as well as corvids worldwide. We detected all three genera of avian haemosporidians in Northwestern Crows in Alaska, although only Leucocytozoon occurred at all sites in both years. Given the strong geographic and interannual variation in parasite prevalence and apparent lack of correlation with climatic variables, it appears that there are other key factors responsible for driving transmission dynamics in this region. Thus, caution is warranted when using standard climatic or geographic attributes in a predictive framework. Our phylogenetic results demonstrate lower host specificity for some lineages of Leucocytozoon than is typically reported and provide insights about genetic diversity of local haemosporidian parasites in Alaska.

Parasites & Vectors

A novel technique for isolating DNA from Tempus™ blood RNA tubes after RNA isolation

Objective We use Tempus blood RNA tubes (Applied Biosystems) during health assessments of American moose ( Alces alces spp.) as a minimally invasive means to obtain RNA. Here we describe a novel protocol to additionally isolate high-quality DNA from the supernatant remaining after the RNA isolation methodology. Metrics used to qualify DNA quality included measuring the concentration, obtaining a DNA integrity number from a genomic DNA ScreenTape assay (Agilent), and running the isolated DNA on an agarose gel. Results Of the 23 samples analyzed, the average DNA concentration was 121 ng/µl (range 4–337 ng/µl) and a genomic DNA ScreenTape assay of seven samples indicated high DNA integrity values for 6 of the 7 samples (range 9.1–9.4 out of 10). Of the DNA sent for genotyping by sequencing, all proved to be of sufficient integrity to yield high-quality next-generation sequence results. We recommend this simple procedure to maximize the yield of both RNA and DNA from blood samples.

BMC Research Notes

Taqman Real-Time PCR Detects Avipoxvirus DNA in Blood of Hawaìi `Amakihi ( Hemignathus virens )

Background Avipoxvirus sp. is a significant threat to endemic bird populations on several groups of islands worldwide, including Hawaìi, the Galapagos Islands, and the Canary Islands. Accurate identification and genotyping of Avipoxvirus is critical to the study of this disease and how it interacts with other pathogens, but currently available methods rely on invasive sampling of pox-like lesions and may be especially harmful in smaller birds. Methodology/Principal Findings Here, we present a nested TaqMan Real-Time PCR for the detection of the Avipoxvirus 4b core protein gene in archived blood samples from Hawaiian birds. The method was successful in amplifying Avipoxvirus DNA from packed blood cells of one of seven Hawaiian honeycreepers with confirmed Avipoxvirus infections and 13 of 28 Hawaìi `amakihi ( Hemignathus virens ) with suspected Avipoxvirus infections based on the presence of pox-like lesions. Mixed genotype infections have not previously been documented in Hawaìi but were observed in two individuals in this study. Conclusions/Significance We anticipate that this method will be applicable to other closely related strains of Avipoxvirus and will become an important and useful tool in global studies of the epidemiology of Avipoxvirus .

Hawaii

Effects of nonlethal sea lamprey attack on the blood chemistry of lake trout

A laboratory study examined changes in the blood chemistry of field-caught and hatchery-reared lake trout Salvelinus namaycush subjected to a nonlethal attack by sea lampreys Petromyzon marinus. We measured glucose, total protein, amylase, alkaline phosphatase (ALKP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatine kinase, calcium, magnesium, triglycerides, sodium, and potassium with a Kodak Ektachem DT60 Analyzer, Ektachem DTSC Module, and the DTE Module. Mean levels of total protein, AST, ALKP, hematocrit, calcium, magnesium, and sodium decreased significantly ( P ≤ 0.05), and mean levels of ALT and potassium increased significantly ( P ≤ 0.05) after sea lamprey feeding. Lake trout condition ( K ) and hematocrit levels also decreased significantly ( P ≤ 0.05) after the sea lamprey attack. Frequency distributions of eight lake trout blood chemistry variables and the hematocrit were significantly different before and after a sea lamprey attack. A second study that used hatchery lake trout broodstock measured changes in hematocrit before and after a sea lamprey attack.

Michigan

Evidence for an amoeba-like infectious stage of ichthyophonus sp. and description of a circulating blood stage: a probable mechanism for dispersal within the fish host

Small amoeboid cells, believed to be the infectious stage of Ichthyophonus sp., were observed in the bolus (stomach contents) and tunica propria (stomach wall) of Pacific staghorn sculpins and rainbow trout shortly after they ingested Ichthyophonus sp.–infected tissues. By 24–48 hr post-exposure (PE) the parasite morphed from the classically reported multinucleate thick walled schizonts to 2 distinct cell types, i.e., a larger multinucleate amoeboid cell surrounded by a narrow translucent zone and a smaller spherical cell surrounded by a “halo” and resembling a small schizont. Both cell types also appeared in the tunica propria, indicating that they had recently penetrated the columnar epithelium of the stomach. No Ichthyophonus sp. pseudo-hyphae (“germination tubes”) were observed in the bolus or penetrating the stomach wall. Simultaneously, Ichthyophonus sp. was isolated in vitro from aortic blood, which was consistently positive from 6 to 144 hr PE, then only intermittently for the next 4 wk. Small PAS-positive cells observed in blood cultures grew into colonies consisting of non-septate tubules (pseudo-hyphae) terminating in multinucleated knob-like apices similar to those seen in organ explant cultures. Organ explants were culture positive every day; however, typical Ichthyophonus sp. schizonts were not observed histologically until 20–25 days PE. From 20 to 60 days PE, schizont diameter increased from ≤25 μm to ≥82 μm. Based on the data presented herein, we are confident that we have resolved the life cycle of Ichthyophonus sp. within the piscivorous host.

Journal of Parasitology

Survey of selected pathogens and blood parameters of northern yellowstone elk: Wolf sanitation effect implications

The restoration or conservation of predators could reduce seroprevalences of certain diseases in prey if predation selectively removes animals exhibiting clinical signs. We assessed disease seroprevalences and blood parameters of 115 adult female elk (Cervus elaphus) wintering on the northern range of Yellowstone National Park [YNP] during 2000-2005 and compared them to data collected prior to wolf (Canis lupus) restoration (WR) in 1995 and to two other herds in Montana to assess this prediction. Blood parameters were generally within two standard deviations of the means observed in other Montana herds (Gravelly-Snowcrest [GS] and Garnet Mountain [GM]), but Yellowstone elk had higher seroprevalences of parainfluenza-3 virus (95% CI YNP = 61.1-78.6, GS = 30.3-46.5) and bovine-virus-diarrhea virus type 1 (95% CI YNP = 15.9-31.9, GM = 0). In comparisons between pre-wolf restoration [pre-WR] (i.e., prior to 1995) seroprevalences with those post-wolf restoration [post-WR] in Yellowstone, we found lower seroprevalences for some disease-causing agents post-wolf restoration (e.g., bovine-virus-diarrhea virus type-1 [95% CI pre-WR = 73.1-86.3, post-WR = 15.9-31.9] and bovine-respiratory syncytial virus [95% CI pre-WR = 70.0-83.8, post-WR = 0]), but similar (e.g., Brucella abortus [95% CI pre-WR = 0-4.45, post-WR = 0-4.74] and epizootic hemorrhagic disease virus [95% CI pre-WR = 0, post-WR = 0]) or higher for others (e.g., Anaplasma marginale [95% CI pre-WR = 0, post-WR = 18.5-38.7] and Leptospira spp. [95% CI pre-WR = 0.5-6.5, post-WR = 9.5-23.5]). Though we did not detect an overall strong predation effect through reduced disease seroprevalence using retrospective comparisons with sparse data, our reference values will facilitate future assessments of this issue.

American Midland Naturalist

Absorption of p,p'-dichlorodiphenyldichloroethylene and dieldrin in largemouth bass from a 60-D slow-release pellet and detection using a novel enzyme-linked immunosorbent assay method for blood plasma

This work describes the uptake of two organochlorine pesticides from slow-release pellets by largemouth bass and the utility of a blood plasma enzyme-linked immunosorbent assay (ELISA) method for exposure verification. We measured blood and tissue levels by gas chromatography/mass spectrometry and by a novel ELISA method, and present a critical comparison of the results.

Environmental Toxicology and Chemistry

Blood analyses of wolf pups and their ecological and metabolic interpretation

Blood samples were obtained from 32 wolf ( Canis lupus ) pups live-trapped over a three-year period in northern Minnesota. The results of 21 laboratory analyses of hematology and blood chemistry are tabulated and analyzed in terms of study area, age, sex, and year of collection. Mean values are compared to those reported for dogs in the same age group. The numerous differences between dog and wolf pups are interpreted in terms of nutritional levels and dietary composition with the suggestion that the wolves are not achieving their full growth potential. Individual abnormal test results are tabulated and possible interpretations are suggested. Abnormal results were observed in 13 animals including 10 of 11 animals sampled in 1972. The results in the 1972 animals indicated a poorer nutrition. This preponderance of abnormal test results in pups from 1972 is correlated with ecological studies on this wolf population indicating decreased survival. The potential value of such long-term integrated field and laboratory studies for providing a more complete understanding of changes in the dynamics of natural populations in terms of the responses of individual animals is demonstrated.

Minnesota

Effects of winter fasting and refeeding on white-tailed deer blood profiles

This study examined the effects of dietary protein, fasting, and refeeding on blood characteristics of 9 nonpregnant, female white-tailed deer ( Odocoileus virginianus ) in captivity from 23 February to 3 May 1984. Percent weight loss was greater in fasted deer than in deer fed diets of 2 crude protein levels. Fasting effects were also observed for hemoglobin (Hb), red blood cell (RBC) counts, packed cell volume (PCV), cholesterol, triglycerides, serum urea nitrogen (SUN), potassium (K), glucose, phosphorus (P), insulin, thyroxine (T4), and total protein (TP). Refeeding influenced cholesterol, sodium (Na), and calcium (Ca). Hemoglobin, PCV, Ca, P, and albumin varied with time in fasted deer. Changes over time in the fed deer occurred for several hematological and serum characteristics. Data are presented to serve as reference values for better understanding of data collected from free-ranging deer under less known conditions.

Minnesota

Concentrations of Elements in Hellbender Blood and Fish Fillets from the Missouri Department of Conservation Monitoring Programs

This report presents the results of contaminant monitoring surveys conducted annually by the Missouri Department of Conservation to examine the levels of selected elemental contaminants in hellbender (Cryptobranchus alleganiensis) blood and fish. Catfish (Ictalurus furcatus, Ictalurus punctatus, Pylodictis olivaris), redhorse (Moxostoma anisorum, Moxostoma erythrurum), bass (Micropterus salmoides, Micropterus punctulatus, Micropterus Lacepede, Ambloplites rupestris), walleye (Sander vitreus), and sunfish (Lepomis megalotis) were collected from 17 sites as part of the Department's General Contaminant Monitoring Program. Bluegill (Lepomis macrochirus) and other sunfish (Lepomis megalotis, Lepomis cyanellus) were collected from 18 sites as part of the Department's Resource Assessment and Monitoring Program. Blood from hellbenders was collected from seven sites as part of the Department's Hellbender Monitoring Program.

Open-File Report

Sex and storage affect cholinesterase activity in blood plasma of Japanese quail

Freezing at -25?C had confounding effects on cholinesterase (ChE) activity in blood plasma from breeding female quail, but did not affect ChE activity in plasma from males. Plasma ChE activity of control females increased consistently during 28 days of storage while both carbamate- and cidrotophos-inhibited ChE decreased. Refrigeration of plasma at 4?C for 2 days had little effect of ChE activity. Plasma ChE activity was averaged about 34% higher in breeding males than in females. Extreme caution should be exercised in use of blood plasma for evaluation of anti ChE exposure in free-living birds.

Journal of Wildlife Diseases

Blood types in Pacific salmon

Intraspecific differences in erythrocyte antigens (blood types) were shown to occur in four species of Pacific salmon, the sockeye or red salmon ( Oncorhynchus nerka ), the chinook or king salmon ( 0. tshawytsch a), the chum salmon ( O. keta ), and the pink salmon ( O. gorbuscha ). Antisalmon-erythrocyte sera prepared in rabbits and chickens were used after absorption of species-specific antibodies. Some of these blood types were shown to differ in their frequency of occurrence between different geographic races. In addition, isoimmunizations were conducted on one race of sockeye salmon. Antisera of seven different specificities were prepared and at least eight different patterns of antigenic composition were displayed by the cells tested. These results indicate that considerable antigenic diversity exists in salmon. Reagents to detect valuable markers for the investigation of geographic races of salmon should be obtained through further research.

International North Pacific Fisheries Commission B

Use of whole blood samples preserved in DNA lysis buffer for serological detection of avian malaria in Hawaiian forest birds

Recent detections of avian malarial parasites in native and non-native forest birds at Hakalau Forest National Wildlife Refuge and reports of epidemic transmission of the disease in high elevation habitats as well as controversy over accuracy of the PCR (polymerase chain reaction) diagnostic test that was being used led to a request by U.S. Fish and Wildlife Service to see if existing blood samples that were preserved in a DNA lysis buffer could be used for independent confirmation of the findings with antibody based serological methods. The primary objective of this study was to test whether some DNA buffers used for preservation of blood samples cause denaturation and loss of antigenicity of antibody molecules. If the buffer does not destroy antigenicity of these molecules, then the samples can be used in serological assays to provide an independent assessment of the accuracy of PCR tests.

Hawaii

Analysis of cyanide in whole blood of dosed cathartids

A gas-liquid chromatographic method was modified to quantify both unmetabolized (“free”) and metabolized (“bound”, i.e., thiocyanates) cyanides. The methods for both are efficient and sensitive to 0.05 ppm. Repeated freezing and thawing of whole blood from treated cathartids caused an initial increase in free cyanide concentrations, followed by a gradual decline to a plateau. Bound cyanide concentrations declined after repeated freezing and thawing.

Environmental Toxicology and Chemistry

Use of blood levels to infer carcass levels of contaminants

Inferences may be made about the carcass levels of a contaminant based on the contaminant level in blood samples. A method is given for comparing such populations that utilizes bivariate normal distributions and their principal axes, thereby avoiding a dilemma arising from the use of regression techniques. Confidence intervals and power calculations are given. Data from captive barn owls provide partial justification for the use of this method.

Archives of Environmental Contamination and Toxico