Nucleotide sequence variants of Rattus norvegicus mitochondrial DNA

The mitochondrial DNA (mtDNA) molecules of different albino, domesticated rats (Rattus norvegicus) of the SASCO colony are of two kinds (SASCO-1 and SASCO-2) in regard to their sensitivity at certain sites to a number of restriction enzymes. MtDNA molecules from Utah wild R. norvegicus (Wild-UT) have sensitivities to restriction enzymes which differ at some sites from either SASCO-1 or SASCO-2 mtDNA molecules. Four single nucleotide differences were found among the HindIII F fragments (169 nucleotides) of SASCO-1, SASCO-2, and Wild-UT mtDNAs. Arguments are presented in favor of the interpretation that each variant nucleotide is the third nucleotide of the codon containing it, and that none of the four differences would result in a difference in the respective amino acid translated.Dedicated to Professor W. Beermann on the occasion of his 60th birthday     

The impact of surface mine reclamation on headwater streams in Southwest Virginia

Recovery of headwater streams following the cessation of mining and the application of terrestrial (vegetative) reclamation techniques was assessed by comparison of water quality and aquatic biota in two such systems (reclaimed four to seven years) with that of an unimpacted stream and of streams draining mine areas which were abandoned without reclamation. Alkalinity, hardness, sulfate, and conductivity were elevated in the reclaimed mine streams as were fine-particle suspended solids and sediment. Overall water quality was comparable to streams draining unreclaimed lands. Benthic invertebrate and fish populations were significantly lower in abundance in the reclaimed mine streams than in the reference stream and showed less taxonomic richness and stability; they were similar in these respects to the biota of the unreclaimed mine streams. Continued sedimentation from mined areas and haul roads affected stream habitat and appeared to be the major factor limiting biotic recovery. These findings emphasize that terrestrial reclamation does not assure lotic restoration. Water quality criteria merit consideration in the refinement of reclamation procedures for mined lands.     

Monitoring river periphyton with artificial benthic substrates

The objective of this research was to identify the materials and methods necessary to study the attached algal community on a river bottom in deep water. The study site was the Susquehanna River near Falls, Pennsylvania. Artificial substrates of smooth glass, frosted glass, Vermont slate, sandy slate (flagstone) and acrylic plate were placed on the stream bottom in detritus free sample holders by scuba divers. Both monthly and long-term cumulative samples were collected from the plates employing scuba and a Bar-Clamp sampler. River stones (natural substrates) were collected for comparison. Samples were analyzed in a Palmer Cell under a Bausch and Lomb research microscope. Diatoms were the most important colonizers of river stones, with the genera Nitzschia and Navicula most abundant. Highest periphyton densities occurred on natural substrates in winter with a maximum of 2.2 × 10⁴ units/ mm². Artificial substrates with one month exposure periods accumulated maximum periphyton density from May through October with relatively low densities in winter. Cumulative artificial substrates were most like river stones in patterns of colonization. Frosted acrylic is recommended for future studies employing benthic artificial periphyton substrates.This study was partially supported by the Pennsylvania Power and Light Company     

Population dynamics of daphnia pulex and utilization by the rainbow trout (salmo gairdneri)

The effect of rainbow trout (Salmo gairdneri) predation on the population dynamics of the water flea,Daphnia pulex, was examined during 1976 and 1977 in Becker Lake, a small, shallow, productive reservoir in northeastern Arizona.Rainbow trout were size-biased feeders, utilizing daphnids which were 1.3 mm in size or larger. Trout predation uponDaphnia pulex occurred mainly during winter and early spring when their numbers were relatively low but their clutch size high, suggesting that trout selectedDaphnia on the basis of brood pigmentation. By far the greatest proportion ofDaphnia mortality was due to nonpredatory sources, since generally less than 6% could be attributed to trout predation.TheD. pulex population exhibited a bimodal seasonal abundance curve which was attributed to ephippial egg production and trout predation during the winter and poor food quality/quantity during the summer.This work was supported in part by the Arizona Department of Game and Fish. The authors wish to thank Jim Novy and Joe Stone of that department for their invaluable assistance in the field collecting segment of this study.     

Thermoregulatory behavior of juvenile cromileptes altivelis (serranidae), a tropical marine fish

Ten juvenile Cromileptes altivelis were tested individually for 3-day periods in electronic shuttleboxes to measure their thermoregulatory behavior. The fish voluntarily occupied a 21–27°C span, out of a potentially available 0–50°C range. The mean final thermal preferendum was 24.5°C for all individuals. The thermoregulatory performance of this species is similar to that of cool temperate freshwater fishes.     

Complement-dependent induction of DNA synthesis and proliferation of human diploid fibroblasts

The effects of fresh human serum (FHS) and heat-inactivated human serum (HHS) on the DNA synthesis and proliferation of human diploid fibroblasts were assessed. FHS activated significantly more quiescent fibroblasts to undergo DNA synthesis and proliferation than did HHS. The stimulatory effect occurred consistently over a serum concentration range of 0.1–10%. Using bromodeoxyuridine selective killing techniques, it was shown that this FHS stimulatory effect was on a specific subpopulation of fibroblasts unresponsive to HHS. The involvement of the complement system, and specifically of C1, was shown by the inability of Clq-depleted FHS to support enhanced DNA synthesis whereas Clq-depleted serum reconstituted with purified Clq was effective. Purified Clq did not restore activity when added to heated serum, nor was it mitogenic when tested in basal medium without serum. The addition of purified Clq to fresh serum inhibited the enhancement of DNA synthesis, and at Clq concentrations of 4γ/ml and greater, the fresh serum effects were abrogated. Thus, it appears that binding of the assembled C1 complex to the fibroblast surface was required for FHS-mediated enhancement of fibroblast proliferation, with Clq subcomponent serving as the recognition site. The results from several experiments indicated that antibody was not required for the complement-dependent fibroblast activation. FHS was not cytotoxic, and autologous serum was as effective as allogeneic sera. A 20-fold molar excess of Fab' from pooled human IgG did not alter the FHS effects. FHS from which IgG was more than 99% depleted was still effective. These results suggested an antibody-independent role for complement in the activation of a subpopulation of human diploid fibroblasts.     

Cytochalasin B blocks sperm incorporation but allows activation of the sea urchin egg

Cytochalasin B (CB) (2 × 10⁻⁶ M) prevents the incorporation of sperm into the eggs of Lytechinus pictus and Strongylocentrotus purpuratus as judged by light and transmission electron microscopy (TEM). At lower concentrations of CB (2 × 10⁻⁷ M), sperm are successfully incorporated into the egg, but their migration in the area of the egg cortex is impaired. The site of action of CB on the sperm may be on the initial rotation of the sperm nucleus in the cortex; the subsequent migration is not affected by CB. Although sperm incorporation is prevented at the higher CB concentrations, the eggs become activated—as judged by cortical reaction, increased protein synthesis and increased respiration. These findings raise the concept that egg activation by sperm could result from some pre-fusion event and hence that sperm-egg fusion would not be a prerequisite for the triggering of development. An alternative hypothesis is that fusion occurs between the acrosome process membrane and egg membrane, but since CB has destroyed the integrity of the cortex actin, the fusion bridge is so weak that it cannot be maintained without some contractile or cytoskeletal support by the cortex. The sperm may activate the CB-treated egg in the same manner as pricking with a microelectrode sometimes does.     

Evaluation of Bone Strength During Aflatoxicosis and Ochratoxicosis

Young chickens were fed graded levels of aflatoxin (0, 0.625, 1.25, 2.5, 5.0, and 10.0 μg/g of diet) or ochratoxin (0, 0.5, 1.0, 2.0, 4.0, and 8.0 μg/g of diet), and the breaking strength, displacement before failure, and diameter of their tibias were determined. Breaking strength was decreased at growth inhibitory levels of aflatoxin (2.5 μg/g) and ochratoxin (2 μg/g), whereas a reduction in diameter required higher levels (5.0 and 4.0 μg/g, respectively). Bones from birds with ochratoxicosis selected to have diameters equal to control bones had lower breaking strength. In an attempt to negate mathematically the effect of decreased diameter and bias in any selection process, stress at time of failure of the bones was calculated and found to be decreased by feeding aflatoxin but not ochratoxin. Total displacement of bones before breaking was increased significantly (P < 0.05) by both toxins at the highest levels administered, but this increase was primarily the result of an increase in displacement from the start of failure to complete failure. Increased displacement associated with both toxicoses was equal in bones selected to be of equal diameter or in bones from the same treatment but of different diameters. However, calculation of modulus of elasticity which is corrected for diameter revealed aflatoxin had no effect whereas ochratoxin tripled the effect. These data indicate that the material properties of bones can be altered during mycotoxicoses and suggest yet another way in which mycotoxins are detrimental to animal health.     

Fluorometric assay for 1-deoxyfructose

A sensitive fluorometric assay has been developed for the measurement of 1-deoxyfructose in biological fluids. Samples containing 1-deoxyfructose are incubated with an equal volume of 0.01 m 3,5-diaminobenzoic acid in 5.0 m phosphoric acid in a boiling-water bath for 15 min. The fluorescent product has an emission maximum at 502 nm and an excitation maximum at 396 nm. Fluorescence is proportional to 1-deoxyfructose concentrations over a range from 0.002 to 1.0 mm. The method can be used to detect as little as 0.03 μmol of 1-deoxyfructose in deproteinized blood and in urine and no interference is observed with glucose, fructose, pyruvate, or ketone bodies. The method will also detect 1-deoxytagatose, 2-deoxyaldohexoses, and -pentoses, 2,5-anhydromannose, and a number of 2-, 3-, 4-, and 5-mono- or bis(hydroxymethyl)furans. The fluorescence properties of the products formed from all of the above compounds are similar suggesting structural similarities of the adducts formed and possible mechanistic similarities of the reactions involved.