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A rapid field techniques for preparing ant chromosomes for karyotypic analysis

This technique for chromosomal preparation of ant tissues for karyotypic analysis is advantageous under field conditions because it reduces processing time and can be used under humid conditions. The cerebral ganglia from prepupae or early pupae are incubated 20 minutes in a hypotonic citrate solution, minced in a fixative solution of 3:3:4 glacial acetic acid: absolute methanol: distilled HOH, rinsed in a fixative solution of 1:1 glacial acetic acid: methanol followed by Carnoy's fixative, then immediately flame dried. The resulting metaphase chromosomes are well spaced and usually show banding characteristics.

Stain Technology

Transport of microspheres and indigenous bacteria through a sandy aquifer: Results of natural- and forced-gradient tracer experiments

Transport of indigenous bacteria through sandy aquifer sediments was investigated in forced- and natural-gradient tracer teste. A diverse population of bacteria was collected and concentrated from groundwater at the site, stained with a DNA-specific fluorochrome, and injected back into the aquifer. Included with the injectate were a conservative tracer (Br- or Cl-) and bacteria-sized (0.2-1.3-??m) microspheres having carboxylated, carbonyl, or neutral surfaces. Transport of stained bacteria and all types and size classes of microspheres was evident. In the natural-gradient test, both surface characteristics and size of microspheres affected attenuation. Surface characteristics had the greatest effect upon retardation. Peak break-through of DAPI-stained bacteria (forced-gradient experiment) occurred well in advance of bromide at the more distal sampler. Transport behavior of bacteria was substantially different from that of carboxylated microspheres of comparable size.

Environmental Science & Technology

Establishing genome sizes of focal fishery and aquaculture species along Baja California, Mexico

Genome size—the total haploid content of nuclear DNA— is constant in all cells in individuals within a species, but differs among species. Consequently, the genome size is a quantifiable genetic signature that not only characterizes a species, but it can reflect chromatin modifications, which play fundamental roles in most biological processes that are involved in the manipulation and expression of DNA. This characteristic makes the genome size a crucial parameter for genetic research on endemic aquatic species and for genetic manipulations in aquaculture species. Technologies for genetic assessments and improvements applied to fishery and aquaculture species use genome size values as a means by which hybrids, polyploids, and sex can be identified, when sex chromosomes exist. The objectives of this study were to determine genome sizes of aquatic species with economic and biological importance along the Pacific coast of Mexico, as well as to identify the appropriate reference standards for use in this study. Blood, hemolymph or milt were collected from 10 species occurring along the coast of Baja California: Sablefish Anoplopoma fimbria , Black Snapper Lutjanus novemfasciatus , California Halibut Paralichthys californicus , Pacific Sardine Sardinops sagax , Flag Rockfish Sebastes rubrivinctus , Starry Rockfish Sebastes constellatus, Totoaba Totoaba macdonaldi , Whiteleg Shrimp Litopenaeus vannamei and two Yellowtail Seriola lalandi and S. dorsalis . Nuclear DNA was stained with propidium iodide solution and the genome size was determined by flow cytometry, with results ranging from 0.61 pg (1.22 pg/diploid cell) to 2.59 pg (5.18 pg/diploid cell), with the smallest value in Sablefish and the largest in the Whiteleg Shrimp. No significant differences were detected ( P ≤ 0.05) among individuals of the same species; the likely reason behind any dissimilar DNA content values with those from the literature were differences in methodologies or variations in genetics. Red-ear Slider Turtle Trachemys scripta elegans 2.65 pg (5.30 pg/diploid cell) and Red Junglefowl Gallus gallus 1.27 pg (2.54 pg/diploid cell) were chosen as the standards for reference values. These results establish the basis for the Mexican National Aquatic Genetic Resources project supporting genetic improvements for aquaculture and conservation status parameters for fisheries species.

Baja California

Cytochrome P450 1A expression in midwater fishes: Potential effects of chemical contaminants in remote oceanic zones

Cytochrome P450 1A (CYP1A) induction is a robust marker for exposure to polynuclear aromatic hydrocarbons and planar halogenated aromatic hydrocarbons that are aryl hydrocarbon receptor agonists. We examined CYP1A expression in mesopelagic fishes from the western North Atlantic. Individuals in 22 species were obtained from slope water and the Sargasso Sea in 1977, 1978, and 1993. Aryl hydrocarbon hydroxylase (AHH), a CYP1A activity, was detected in liver from all species in 1977/78. In some, including Gonostoma elongatum, AHH was inhibited by the CYP1A inhibitor ??-naphthoflavone. CYP1A-dependent ethoxyresorufin O-deethylase (EROD) was detected in liver microsomes of all species in 1993; rates were highest in G. elongatum and Argyropelecus aculeatus. Immunoblot analysis with the CYP1A-specific monoclonal antibody 1-12-3 detected a single microsomal protein band in most 1993 samples; the highest content was in G. elongatum. Immunohistochemical analysis showed CYP1A staining in gill, heart, kidney, and/or liver of several species. Extracts of the 1993 G. elongatum and A. aculeatus, when applied to fish hepatoma cells (PLHC-1) in culture, elicited a significant induction of EROD in those cells. The capacity of the extracts to induce CYP1A correlated with the content of PCBs measured in the same fish (2-4.6 ng/g total body weight). Mesopelagic fish in the western North Atlantic, which experience no direct exposure to surface waters or sediments, are exposed chronically to inducers of CYP1A at levels that appear to be biochemically active in those fish.Cytochrome P450 1A (CYP1A) induction is a robust marker for exposure to polynuclear aromatic hydrocarbons and planar halogenated aromatic hydrocarbons that are awl hydrocarbon receptor agonists. We examined CYP1A expression in mesopelagic fishes from the western North Atlantic. Individuals in 22 species were obtained from slope water and the Sargasso Sea in 1977, 1978, and 1993. Aryl hydrocarbon hydroxylase (AHH), a CYP1A activity, was detected in liver from all species in 1977/78. In some, including Gonostoma elongatum, AHH was inhibited by the CYP1A inhibitor ??-naphthoflavone. CYP1A-dependent ethoxyresorufin O-deethylase (EROD) was detected in liver microsomes of all species in 1993; rates were highest in G. elongatum and Argyropelecus aculeatus. Immunoblot analysis with the CYP1A-specific monoclonal antibody 1-12-3 detected a single microsomal protein band in most 1993 samples; the highest content was in G. elongatum. Immunohistochemical analysis showed CYP1A staining in gill, heart, kidney, and/or liver of several species. Extracts of the 1993 G. elongatum and A. aculeatus, when applied to fish hepatoma cells (PLHC-1) in culture, elicited a significant induction of EROD in those cells. The capacity of the extracts to induce CYP1A correlated with the content of PCBs measured in the same fish (2-4.6 ng/g total body weight). Mesopelagic fish in the western North Atlantic, which experience no direct exposure to surface waters or sediments, are exposed chronically to inducers of CYP1A at levels that appear to be biochemically active in those fish.

Environmental Science & Technology