An analysis of the micronuclei in the oral mucosa for assessing the cytogenetic effect of environmental pollutants

The frequencies of micronuclei in the buccal mucosa cells are investigated in several groups: in the control group and in the groups of workers from the pure iron factory (populated area A): in the control group and in the group of workers from the microbiological works (area B). Frequencies of micronuclei are shown to somewhat increase in workers of both works as compared to the control groups. Levels of micronuclei are revealed to increase in the smokers as against the nonsmokers in the group of workers from the microbiological factory. The above examination permits recommending analysis of micronuclei to estimate xenobiotics polluting respiratory tracts.     

The evolution of the major histocompatibility complex in upstream versus downstream river populations of the longnose dace

Populations in upstream versus downstream river locations can be exposed to vastly different environmental and ecological conditions and can thus harbor different genetic resources due to selection and neutral processes. An interesting question is how upstream–downstream directionality in rivers affects the evolution of immune response genes. We used next‐generation amplicon sequencing to identify eight alleles of the major histocompatibility complex (MHC) class II β exon 2 in the cyprinid longnose dace (Rhinichthys cataractae) from three rivers in Alberta, upstream and downstream of municipal and agricultural areas along contaminant gradients. We used these data to test for directional and balancing selection on the MHC. We also genotyped microsatellite loci to examine neutral population processes in this system. We found evidence for balancing selection on the MHC in the form of increased nonsynonymous variation relative to neutral expectations, and selection occurred at more amino acid residues upstream than downstream in two rivers. We found this pattern despite no population structure or isolation by distance, based on microsatellite data, at these sites. Overall, our results suggest that MHC evolution is driven by upstream–downstream directionality in fish inhabiting this system.     

DNA Damage and Repair in Stylonychia lemnae (Ciliata,Protozoa)1

ABSTRACT Irradiation with X rays, UV irradiation after incorporation of bromodeoxyuridine (BU) into the DNA, and cis-platinum (cis-Pt) treatment each cause the loss of micronuclei of Stylonychia lemnae while the macronuclei are not severely affected. The abilities of both nuclei to repair DNA were investigated. Unscheduled DNA synthesis could not be demonstrated after X-ray irradiation, but it was found after treatment with BU/UV and cis-Pt in macro- and micronuclei. The extent of the repair process in the micro- and macronuclei was alike, as indicated by grain counts of [6-³H]thymidine-treated cells. One reason for the different sensitivity of both nuclei to DNA-damaging treatment may be the different number of gene copies in the macro- and micronuclei.     

Effects of clay discharges on streams

The impact of clay discharges on benthic invertebrates was investigated by comparison of communities upstream and downstream of alluvial gold mining on 6 streams on the West Coast of the South Island, New Zealand. Mean turbidity was increased by 7–154 NTU above background (mean 1.3–8.2 NTU) by the mine discharges during the 2 months before sampling. Patterns of increase in suspended solids (strongly correlated with turbidity, r=0.95) were similar. Invertebrates densities were significantly lower at all downstream sites, ranging from 9 to 45% (median 26%) of densities at matched upstream sites. Downstream densities as a proportion of those upstream were negatively correlated with the logarithm of the turbidity loading (r=–0.82, P<0.05). The densities of the common taxa were also generally lower downstream of mining. Taxonomic richness was significantly lower at downstream sites in the four streams receiving higher turbidity loads (mean turbidity increase = 23–154 NTU). Lower epilithon biomass and productivity, and degraded food quality at the downstream sites probably explain the lowered invertebrate densities. At some sites, reduced bed permeability and interstitial dissolved oxygen, and avoidance reactions of invertebrates (i.e., increased drift), may have also contributed to lower invertebrate densities.Formerly DSIR Marine and Freshwater.     

Impact of an urban effluent on the bacterial community structure in the Arga River (Spain), with special reference to culturable gram-negative rods

The Arga River is an interesting system in which to study the impact of urban effluent pollution because it receives a single effluent in the form of wastewater discharge from the city of Pamplona. To analyze the extent of this discharge, total bacteria, culturable heterotrophic bacteria, and Gram-negative heterotrophic bacteria were enumerated and 409 isolates of the latter were identified. One sampling station was located upstream from the inflow, while five were located up to 30 km downstream. Bacterial counts increased drastically downstream from the wastewater inflow. Total bacterial numbers decreased along the 30 km downstream, the last station attaining similar values to those recorded upstream from the discharge. However, culturable heterotrophic and Gram-negative heterotrophic bacteria levels generally remained significantly higher within the 30 km zone investigated. Among the 409 isolates identified, Aeromonas spp. were the most frequent both upstream and downstream from the discharge. In contrast, although strains belonging to different genera of Enterobacteriaceae were found in all stations, their occurrence was significantly higher just downstream from the polluted discharge. Acinetobacter spp., which were never found upstream, were detected in all stations below the discharge. Our results clearly show that the bacterial community structure changes definitively downstream from the discharge and that Aeromonas were common throughout the sampling zone. Thus they cannot be considered good indicators of pollution in this setting compared to some genera of Enterobacteriaceae or some species of Acinetobacter, the distribution of which correlated better with the distance from the wastewater discharge.     

Spatial and temporal variation of fish assemblages and their associations to habitat variables in a mountain stream of north Tiaoxi River,China

The spatial and temporal variations of the fish assemblages in mountain streams of China are poorly understood. The relationships between the fish assemblage and selected habitat features were examined in the North Tiaoxi River, one of headwaters of Taihu Lake. A total of 3,348 individuals belonging to 5 orders, 11 families, 25 genera and 34 species were collected including 33 native species and one invasive species. Among those, about 20 species were endemic to China. Non-metric Multidimensional Scaling (NMDS) was applied to compare fish assemblage structures from upstream to downstream during four seasons. Species assemblages differed along the stream continuum, but there was little apparent change associated with the seasons. Species richness and Shannon-Weaver index (H′) tended to increase along the stream continuum from the upstream to downstream and the proportion of invertivorous fish tended to significantly decrease along the continuum with a parallel significant increase in the percentage of omnivores. Fish assemblages were significantly related to both water quality and habitat structure variables. Canonical Correspondence Analysis ordinations (CCA) revealed that 6 of the 14 selected environmental variables had significant relationships with the fish assemblage such as distance to source, stream width, altitude, pH, water depth, and water velocity and different sampling sites were associated with different environmental variables in different seasons. The main differences in fish assemblage structure and diversity within the whole watercourse are probably related to large-scale factors such distance to source, altitude and stream width. Differences of instream characteristics are likely to be caused by natural variability of the ecosystems but also, in some case, by anthropogenic influence like human settlements, agriculture and river embankment and pollution from small factory.     

Upstream-downstream movements of aquatic invertebrates in a Rocky Mountain stream

Simultaneous collections of drift and organisms moving either upstream or downstream in association with the substrate were made using a specially designed sampler. Samples were taken in a diel series along a transect across the study riffle of a Colorado foothills stream on six dates over an annual cycle. In addition to longitudinal movements, taxonomic composition and diel periodicity were evaluated. The insect-dominated fauna showed a net downstream displacement. Only the caddisflies Helicopsyche borealis and Hesperophylax occidentalis exhibited net upstream movement, primarily a result of low drift frequencies. The taxonomic composition of moving invertebrates differed from that of the benthos. Drift resembled downstream moving substrate-associated invertebrates in composition, but differed from that of the upstream directed fauna. Taxa collectively exhibited four types of diel patterns: 1) similar downstream (drift and substrate-associated movements) patterns, which generally differed from the upstream pattern; 2) similar benthic (upstream and downstream) patterns, which differed from that of drift; 3) aperiodic patterns; and 4) independent patterns for each type of directional movement. Analysis of size classes based on head capsule width for the mayfly Baetis tricaudatus showed significantly smaller size in stationary individuals compared with moving individuals in the population and revealed that nymphs moving during the day were smaller than those moving at night.     

Influence of wastewater treatment on the microbial ecology of a large, temperate river system — the Danube River

In an attempt to assess the influence of tertiary-treated sewage influx on bacterial metabolism in the Danube River, bacterial abundance, ectoenzymatic activity, faecal coliforms, faecal streptococci, chlorophyll a (chl a), concentrations of dissolved (DOC) and total organic carbon (TOC) and inorganic nutrients were measured upstream of the sewage influx and compared with sampling sites downstream. Additional samples were taken near the outlet of the sewage treatment plant. Bacterial abundance as determined by epifluorescence microscopy was compared with plate counts of total heterotrophic bacteria. Significantly higher values were obtained at the stations downstream from the sewage influx only for faecal coliforms and faecal streptococci, for glucuronidase activity and bacterial biovolume. All the other parameters were not significantly different from values obtained at the upstream sampling site. Strong seasonal dependence was detectable for nitrate with high concentrations during the winter ( 250 µM) and low concentrations during summer ( 100 µM). A distinct spring phytoplankton bloom was noticeable in the river reaching chl a concentrations of 70 µg 1^–1; during the remaining seasons chl a concentrations were <20 µg 1^–1. Highly significant correlations were found between faecal coliform counts and glucuronidase activity. C:N:P ratios of 13.9:10.7:1 (for the upstream station) and 11.7:9.2:1 (for the downstream stations) of dissolved nutrients are rather low indicating severe carbon limitation of bacterioplankton. Based on our results we conclude that the heterotrophic bacterial community is not significantly effected by the input of treated sewage but faecal contamination is readily detectable over a comparatively long reach of 30 km.     

Induction of kinetochore positive and negative micronuclei in V79 cells by N-methyl-N′-nitro-N-nitrosoguanidine: the protective effect of the pyridoindole antioxidant stobadine

The induction of micronuclei by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and their reduction by the cardioprotective synthetic antioxidant, stobadine were studied in hamster V79 cells cultured in vitro. The micronuclei derived from acentric fragments or from whole chromosomes were evaluated with the help of an immunofluorescent staining using antikinetochore antibodies from the serum of scleroderma (CREST syndrome) patients. Our results showed that MNNG (0.5 μg/ml) induced mainly kinetochore-negative micronuclei. At 6, 24 and 48 h after MNNG treatment, we measured a 2.7-, 4.3- and 7.0-fold increase, respectively, of kinetochore-negative micronuclei over the controls. The increase of kinetochore-positive micronuclei was rather low and represented at 6, 24 and 48 h, respectively 0.9-, 1.8- and 2.6-fold increases over the controls. Stobadine decreased the level of kinetochore-negative micronuclei at 6, 24 and 48 h to approximately one-half; the frequency of kinetochore-positive micronuclei was reduced only at 6 h. We suppose that the antioxidant stobadine reduces the induction of micronuclei by MNNG by scavenging of MNNG-induced highly reactive OH radicals which cause chromosomal damage.     

Anaerobic microbial counts of different potential anaerobic inocula

Sludges of various origin were analysed on the basis of microbial counts of anaerobes in order to investigate potential and alternative sources for inoculating anaerobic digestors. Sludges from neutralization and oil and fat separation tanks of a sugar cane factory, and from a stabilisation pond of a purified terephthalic-acid-producing plant were found to be adequate potential inocula. Activated sludges were found to have both hydrogenotrophic and acetoclastic methanogens and obligate hydrogen-producing acetogens (OHPA). In all sludges tested, hydrogenotrophic methanogens were the dominant strict anaerobes. The ratio between most probable number counts of acetoclastic methanogens and OPHA seems to be different between dispersed sludges and granular sludges. Correspondence to: J. P. Guyot     

The effects of an abandoned roast yard on the fish and macroinvertebrate communities of surrounding beaver ponds

Recently formed beaver-ponds surrounding an abandoned copper-nickel ore roast yard near Sudbury, Ontario, Canada, were influenced by highly acidic and heavy-metal contaminated effluent. Fish, including Culaea inconstans, Phoxinus neogaeus, Phoxinus eos, and Pimephales promelas, were found in the ponds upstream of the roast yard only. Macroinvertebrate taxon richness and diversity were greater in the upstream ponds than in the downstream ponds. Acid intolerant taxa, including Mollusca and Hyalella azteca, were found upstream only; acid tolerant taxa, including Enallagma and Ischnura (Odonata: Coenagrionidae) were found in the downstream ponds. One taxon, a species of Chironomus, was especially tolerant of both acid and metal stresses, and was the only invertebrate taxon found in the pond located directly on the roast yard itself. We conclude that long-abandoned mining and refining sites can continue to have a marked effect on aquatic communities.     

再生水补水对河道底泥细菌群落结构的影响

以北京市永定河麻峪湿地再生水补水口附近河道底泥为研究对象,应用末端限制性片段长度多态性(Terminal Restriction Fragment Length Polymorphism,T-RFLP)技术探究再生水补水口附近底泥细菌群落结构的空间差异特征以及环境因子所产生的影响,并借助典范对应分析(Canonical Correspondence Analysis,CCA)方法分析麻峪湿地底泥细菌群落结构空间差异特征的形成原因。分析结果表明:河道底泥微生物群落对再生水的净化主要发生在距补水口1200m的范围内,随着再生水与上游来水径向混合净化渐变过程,在补水口2000m处河道底泥微生物群落结构与河道再生水补水口上游趋于相似。基于样点聚类结果的群落结构多样性分析表明随着再生水净化过程的实现微生物综合多样性指数呈现下降的趋势,从均匀度指数的变化趋势看,再生水补水口具有最高均匀度指数,补水口下游1200米处由于非优势菌群所占相对丰度最低以及偶见菌群缺失,导致群落结构比较单一、群落多样性和均匀度指数都最低。CCA方面分析结果表明再生水补水口上游细菌群落与因累积效应形成的重金属有密切关系,补水口出口底泥细菌群落则主要受到总磷和总有机碳的影响较大,而再生水补水与上游来水汇水径向渐变过程可能与氨氮净化具密切关系。研究区的主要优势菌种为假单胞菌属、贪食菌属和芽孢杆菌属,是与再生水中有机碳、氮降解具有密切关系的菌属。     

Ecology and population structure of the bayou darter, <Emphasis Type="Italic">Etheostoma rubrum</Emphasis>: disjunct riffle habitats and downstream transport of larvae

The bayou darter, Etheostoma rubrum (Percidae), is endemic to the Bayou Pierre system in Mississippi. Adult and juvenile E. rubrum occupy swift, shallow riffles or runs over coarse gravel and pebble substrata. Habitat requirements of larval and post-larval stages, and the role of downstream dispersal of larvae in colonizing riffles are poorly known. The potential for movement and the high level of habitat specificity for the discontinuous riffle habitat suggest that E. rubrum may comprise a metapopulation of linearly arranged local populations. The greatest population densities of E. rubrum occur in the upper reaches of Bayou Pierre. We hypothesized that metapopulation structure of E. rubrum may include source–sink dynamics, whereby downstream areas are a sink for larvae/early juveniles originating upstream. We tested hypotheses that a transport mechanism, larval drift, occurred in E. rubrum, and that downstream riffles showed characteristics of population sinks. We captured larval stages of E. rubrum in cross-sectional and longitudinal drift nets, and rates of drift tended to increase during the day. Larval E. rubrum (n=19) occurred in samples above and below riffle areas, with no differences among longitudinal drift nets placed above and below riffles. Thus, larval drift is a viable transport mechanism. Relative abundance of adults and juveniles declined from upstream to downstream, and inter-riffle distances increased with cumulative downstream distance. However, both predictions of the source–sink hypothesis were not supported. The distribution of size classes did not change between upstream and downstream riffles nor did the mean size-adjusted body mass.     

Cytogenetic damage in fish (Oncorhynchus mykiss) exposed in situ to water from a polluted river]

Specimens of rainbow trout (Oncorhynchus mykiss) were exposed in situ for 7, 15, and 30 days to the water of the Po River either upstream or downstream the immission of the Lambro River, a small affluent which is highly polluted with industrial wastes. A third group of animals was maintained as control in aquarium with a flow of unchlorinated fresh water. Blood smears were obtained from each of the 90 trout specimens and examined under the microscope. The number of micronucleated erythrocytes (MNE) was registered by scoring at least 2,000 cells per animal. No increases in the number of MNE was detected in the animals exposed upstream the confluence, as compared to controls. Conversely, a significant increase in the frequency of MNE after 7 days of exposure to the water of the downstream site was observed. This frequency remained elevated, yet not significantly, after 15 and 30 days of exposure. This study demonstrates the validity of in situ exposure and the usefulness of cytogenetic end-points for environmental biomonitoring.     

Asymmetric dispersal structures a riverine metapopulation of the freshwater pearl mussel Margaritifera laevis

Unidirectional water flow results in the downstream‐biased, asymmetric dispersal of many riverine organisms. However, little is known of how asymmetric dispersal influences riverine population structure and dynamics, limiting our ability to properly manage riverine organisms. A metapopulation of the freshwater pearl mussel Margaritifera laevis may be sensitive to river currents because mussels are repeatedly exposed to downstream drift during floods—a parasitic life stage is the only, limited period (~40 days) during which larvae (glochidia) can move upstream with the aid of host fish. We hypothesized that water‐mediated dispersal would overwhelm upstream dispersal via host fish, and therefore, that upstream subpopulations play a critical role as immigrant sources. To test this hypothesis, we examined the effects of both up‐ and downstream immigrant sources on the size of target subpopulations in the Shubuto River system, Hokkaido, Japan. We found that target subpopulation size was dependent on the upstream distribution range of reproductive subpopulations and the number of upstream tributaries, which are proxies for the number of potential immigrants moving downstream. In contrast, little influence was observed of downstream immigrant sources (proximity to downstream reproductive subpopulations). These results were consistent even after accounting for local environments and stream size. Our finding suggests that upstream subpopulations can be disproportionately important as immigrant sources when dispersal is strongly asymmetric.     

Coexistence in streams: do source–sink dynamics allow salamanders to persist with fish predators?

Theory suggests that source–sink dynamics can allow coexistence of intraguild predators and prey, but empirical evidence for this coexistence mechanism is limited. We used capture–mark–recapture, genetic methods, and stable isotopes to test whether source–sink dynamics promote coexistence between stream fishes, the intraguild predator, and stream salamanders (Dicamptodon aterrimus), the intraguild prey. Salamander populations from upstream reaches without fish were predicted to maintain or supplement sink populations in downstream reaches with fish. We found instead that downstream reaches with fish were not sinks even though fish consumed salamander larvae—apparent survival, recruitment, and population growth rate did not differ between upstream and downstream reaches. There was also no difference between upstream and downstream reaches in net emigration. We did find that D. aterrimus moved frequently along streams, but believe that this is a response to seasonal habitat changes rather than intraguild predation. Our study provides empirical evidence that local-scale mechanisms are more important than dispersal dynamics to coexistence of streams salamanders and fish. More broadly, it shows the value of empirical data on dispersal and gene flow for distinguishing between local and spatial mechanisms of coexistence.     

Effects of pulsed and pressed disturbances on the benthic invertebrate community following a coal spill in a small stream in northeastern USA

In September 1999, a coal-carrying train derailed and spilled 180,000–270,000 kg of coal into the Cayuga Inlet near Ithaca, New York. This study determined the immediate effects of the coal spill and the clean up procedures on the aquatic invertebrate community, and whether the stream recovered from this event after 2 years. Benthic invertebrate samples were taken both upstream and downstream of the coal spill immediately following clean-up efforts and two years later. Just after the coal spill, the total abundance and species richness of aquatic invertebrates were significantly lower downstream of the spill, suggesting that the disturbance caused increased mortality and/or emigration compared to a reference site upstream. Taxa affected most were grazers and turbidity-susceptible invertebrates. Two years later the invertebrate communities were similar upstream and downstream of the spill, except for an increase in the percent of the dominant genus, Hydropsyche(Trichoptera: Hydropsychidae). We speculate that long-term effects of channelization of the stream that occurred during the clean-up prevented the invertebrate assemblage from returning to the conditions observed in a reference site upstream of the coal spill. We propose that large scale environmental clean-ups should be designed to avoid altering ecosystems permanently, and that streams should be allowed to recover naturally without destructive human intervention.     

Lytic Enzymes for Gram-negative Bacteria Produced by Bacillus subtilis YT-25

The available energy, gross protein value, phosphorus availability and palatability of 16 samples of single cell protein were evaluated in 20 bioassays using total 2,136 depleted chicks.

Four protein samples were products from Aspergillus tamarii grown on waste water of a fish processing factory, three were from Aspergillus oryzae grown on either acetic acid medium or cooked soybean waste, three were from Candida sp. grown on citrus molasses extracted from peel wastes of citrus processing plants, four were from Candida utitis grown on wood molasses produced from various wood wastes, and two were from Pseudomonas sp. and Alteromonas thlasomethanolica grown on methanol.

Five of 16 samples had excellent nutritive value, comparable to single cell proteins available commercially in Europe. All samples were palatable to the chicks, and no sign of acute toxicity was observed.     

Fungal and Bacterial Colonisation of Salix pedicellataLeaves Decaying in Permanent and Intermittent Streams in Eastern Morocco

Leaf degradation was investigated at four sites in the Zegzel/Cherraa river system (Oueds). Two sites (Upper Zegzel and Lower Zegzel) carry water throughout the year and two (Upstream and Downstream Cherraa) are dry for 5–7 months each year. The dynamics of leaf weight loss and microorganisms associated with Salix pedicellata leaves decaying at the four sites were compared for the first time during the same period over this permanent and intermittent system. Overall decay rates of leaves were significantly higher in the permanent Zegzel stream sections (k = 0.0094 d^–1 upstream and 0.0056 downstream) than in the intermittently flowing Cherraa sites (k = 0.0046 upstream, and 0.0036 downstream). In the latter, decay was much slower during dry than during wet periods (Upstream Cherraa: k = 0.0028 and 0.0446, respectively; Downstream Cherraa, k = 0.0008 and 0.0357, respectively). Similar gradients from permanent to intermittent sites were observed in numbers of bacteria per unit area or per weight of decaying leaves (direct counts by epifluorescence microscopy), in numbers of fungal species and in sporulation rates, from leaves recovered at the four sites. Ten hyphomycete species were new for Morocco.