Detection of mining impacts on aquatic macroinvertebrate communities: Results of a disturbance experiment and the design of a multivariate BACIP monitoring programme at Coronation Hill,Northern Territory

Abstract Multivariate methods for the detection of impacts on communities are: (i) explored in the context of a disturbance experiment at a site of previous uranium mining in Kakadu National Park, Northern Territory; and (ii) applied to the design of a monitoring programme for a proposed mining development at nearby Coronation Hill. The disturbance experiment in Rockhole Mine Creek used a gravity-fed redirection of polluted water from the mine adit to create a point-source pollutant. Weekly samples of the macroinvertebrate communities were taken both before and after disturbance at the control and‘impact’ site, as part of a multivariate BACIP (Before-After-Control-Impact-Pairs) design. The results of the experiment show that the multivariate approach provided high sensitivity in detecting impacts on the community, with little difference in sensitivity between species and family level analyses. A simple procedure for deleting those taxa that show poor discrimination between pristine and presently disturbed areas further increased the sensitivity of the tests. In addition to the BACIP analyses, robust ordination of the community data suggested a time/treatment interaction effect. Application of the multivariate BACIP approach also was explored using baseline data for benthic communities from the South Alligator River near Coronation Hill. The method provides high statistical power for monitoring; a relatively small change in community dissimilarity would be detectable in a programme with 5 years of observations before and after commencement of mining. A robust ordination of the yearly samples for the control and potential-impact sites showed that, although samples varied along identifiable gradients over time, the two areas changed in parallel so that the difference between the areas was approximately constant over years. This result demonstrates that the multivariate BACIP approach can provide relative constancy of pre-impact difference values, a key requirement of BACIP designs that is difficult to satisfy using abundance data for a single species.     

DNA fingerprint bands applied to linkage analysis with quantitative trait loci in chickens

Summary
An efficient approach to detect association between quantitative traits and bands of DNA fingerprint patterns uses intra-family tail analysis, which compares fingerprints of DNA mixes from individuals at the two tails of a phenotypic distribution. In analysis of 67 paternal half-sibs of a meat-type chicken family, of 57 sire bands generated by two probes, one sire-specific band (S6–6) was associated with abdominal fat deposition. The band effect was estimated by a linear model analysis to be 0–88 standard deviations, or about 30% of the family mean. The association between band S6–6 and abdominal fat was further examined by testing progeny of paternal half-sibs of the chickens which were used in the tail analysis, establishing genetic linkage between the DNA marker and a genetic locus affecting abdominal fat deposition.     

Normal rat kidney proximal tubule cells in primary and multiple subcultures

Summary Anin vitro model to establish primary and subcultures of rat kidney proximal tubule (RPT) cells is described. After excising the kidneys and separating the cortex, the cortical tissue is digested with the enzyme DNAse-collagenase (Type I) resulting in a high yield of viable RPT Cells. The isolated RPT cells are then seeded onto rat tail collagen-coated surfaces and grown to confluency in a serum-free, hormonally defined medium. The cell yield can be increased by transfering the conditioned medium on Day 1 to more rat tail collagen-coated surfaces. RPT cell attachment and morphology was better on rat tail collagen-coated surfaces than on bovine collagen Type I coated surfaces. The culture medium was a 1∶1 mixture of Ham’s F-12 and Dulbecco’s modified Eagle’s medium supplemented with bovine serum albumin, insulin, transferrin, selenium, hydrocortisone, triiodothyronine, epidermal growth factor, and glutamine. The RPT cells became confluent in 7–10 d, at which point they could be subcultured by trypsinizing and growth in the same medium. In some studies, 10 ng/ml cholera toxin was added to the culture medium. We could passage the RPT cells up to 14 times in the presence of cholera toxin. The cells were investigated for activity of several markers. The cells were histochemically positive for alkaline phosphatase and γ-glutamyl transpeptidase activity and synthesized the intermediate filament pankeratin. The RPT cells displayed apically directed sodium-dependent active glucose transport in culture. Hence, the RPT cells retain structural and functional characteristics of transporting renal epithelia in culture. This rat cell culture model will be a valuable tool for substrate uptake and nephrotoxicity studies.     

Entry of immotile spermatozoa into zona-free hamster ova

We studied six men whose spermatozoa were immotile and possessed a variety of sperm tail structural abnormalities by electron microscopy. The semen of all six subjects had a normal percentage of oval forms and sperm undergoing capacitation and acrosome reaction. Despite the absence of motility, when incubated sperm from these subjects was added to a microdrop of medium containing zona pellucida-free hamster ova, sperm penetration or entry into the cytoplasm of from 1–9% of the eggs was evident with phase contrast microscopy. This latter finding suggests that, at least in this system, oocytes actively facilitate sperm incorporation. Penetration was absent when sperm of fertile men were rendered immotile, though still viable, by heat treatment.     

Tail raising by baboon mothers toward immigrant males

Behavioral responses of adult females to recently immigrated adult males were examined in two groups of chacma baboons in the Moremi Wildlife Reserve, Botswana. Mothers carrying infants respond negatively by raising their tails, avoiding, and screaming when in proximity to immigrant males. These responses are not given by mothers when their infants are away from them or by females who do not have infants. Also, the negative responses by mothers carrying infants are given only to immigrant males and not to long-term resident males. These observations are in concordance with the growing evidence of infanticide in savanna baboon societies: mothers would be expected to show negative responses to potentially aggressive males, particularly immigrants. There was evidence to suggest that mothers respond negatively to immigrants only insofar as these males could not have sired their infants. If confirmed by subsequent observations, this result would lend support to the sexual selection model of infanticide.     

Ecological degradation and agricultural intensification in the Andean highlands

Andean cultural ecologists have made two claims in recent years: ecological decomposition is absent due to effective indigenous management of communal resources, and agricultural intensification is inversely related to altitude. Drawing on material from the Jukumani Indians of Northern Potosi, Bolivia, these assertions are challenged. First, there is little evidence to prove or disprove ecological degradation. Second, the location of agricultural intensification, as the Jukumani data suggests, is influenced by altitude as well as by the presence of market.Fieldwork in Bolivia was carried out between January 1979 and May 1981. This essay was originally presented at a conference entitled, How Communities Resolve Common Property Problems, sponsored by the Harvard Institute for International Development and the Center for Population Studies of Harvard University in the spring semester of 1983.     

Expression of a gene encoding a rabbit sperm membrane protein in mammalian cells.

A general mammalian expression vector designated pSV2-EP was reconstructed by inserting an oligonucleotide fragment into pSV2-dhfr. This vector allowed insertion of cDNAs with EcoRI cohesive ends. The pSV2-EP contains a simian virus 40 (SV40) early promoter, origin for DNA replication, SV40 poly-A site, splicing site, an initiator ATG downstream from the promoter and an EcoRI site for the insertion of cDNA fragment screened from lambda gt11 expression libraries. A recombinant plasmid (pS-VRS-1) was constructed by inserting RSD-1, a cDNA encoding a rabbit sperm tail protein, into the EcoRI site of the pSV2-EP vector. Chinese hamster ovarian (CHO) dhfr-negative cells were cotransformed with pSV2-dhfr and pSVRS-1 by the calcium phosphate method. In selective culture medium without thymidine and hypoxanthine, several cell lines were obtained containing mRNA and DNA that hybridized with RSD-1. One of these transformed cell lines stained intensely with anti-rSMP-B antibodies, demonstrating that the RSD-1 was expressed in the transformed CHO cells.     

Soil restoration under pasture after lignite mining: Management effects on soil biochemical properties and their relationships with herbage yields

The recovery of soil biochemical properties under grazed, grass-clover pasture, after simulated lignite mining, was studied over a 5-year period in a mesic Typic Dystrochrept soil at Waimumu, Southland, New Zealand. The restoration procedures involved four replacement treatments, after A, B, and C horizon materials had been separately removed, from all except the control, and stockpiled for 2–3 weeks. In each replacement treatment, the effects of ripping to 1.8 m depth, mole drainage, and the use of fertilizer nitrogen were also investigated.Replacement treatment markedly influenced the recovery of herbage production and soil organic C and total N contents, N mineralization, microbial biomass (as indicated by mineral-N flush) and invertase and sulphatase activities. The effectiveness of replacement treatments decreased in the order: 1. control (no stripping or replacement). 2 A, B, and C horizon materials replaced in the same order. 3. A, B, and C horizon materials each mixed with an equal amount of siltstone overburden and replaced in order, 4. A and B horizon materials mixed before replacing over C horizon material.Ripping increased herbage production, net N mineralization, and to some extent microbial biomass. Drainage had little, if any, effect.Fertilizer N also stimulated herbage production, but depressed clover growth. Over 2.5 years, it had little detectable effect on the soil properties.Increases in soil invertase and, to a lesser extent, sulphatase activity during the trial were closely related to changes in herbage production. Microbial biomass increased more rapidly than did soil organic C in the early stages of the trial.Rates of net N mineralization strongly suggest that N availability would have limited pasture growth, especially in the treatments with mixed soil materials.