Endangered crayfish in northern Japan: Distribution, abundance and microhabitat specificity in relation to stream and riparian environment

Japan’s only native crayfish species Cambaroides japonicus has been declining dramatically in the past few decades. For the purpose of conservation planning, twenty-two coastal streams were surveyed to investigate summer distributions of crayfish in relation to stream and riparian environment. Classification and regression trees were used to predict the occurrence and abundance of crayfish. The classification tree model with stream variables as predictors showed that crayfish would occur in swift or high gradient streams (correct classification rate = 91%). Within those streams, however, crayfish only inhabited depositional microhabitats, in which the areas are limited in availability. Crayfish were not found in gentle, low gradient streams containing abundant depositional microhabitats. This paradoxical distribution pattern was attributed to availability of boulder substrates in swift or high gradient streams. The regression tree model indicated that crayfish abundance was determined primarily by the percentage of boulder substrates and the presence of fish (observed vs. predicted r = 0.64).The classification tree model using only riparian variables indicated that the total woody plant (mainly broadleaf species) density followed by the percentage of early successional species such as alder and willow determined the splits of the tree model (correct classification rate = 95%). A leaf processing experiment on 10 riparian plant species suggested that crayfish preferred high nitrogen (or low C/N) leaves.These results suggest that swift or high gradient fishless streams associated with abundant cover in dense broadleaf forest serve conservation areas for this endangered crayfish, and that consideration of riparian composition may facilitate conservation efforts.     

Does natural acidity mediate interactions between introduced brown trout, native fish, crayfish and other invertebrates in West Coast New Zealand streams?

The presence of introduced brown trout (Salmo trutta) on the distribution of native crayfish (Paranephrops planifrons), native galaxiid fishes (Galaxias spp.) and invertebrate fauna was investigated in 18 West Coast New Zealand streams (8 with trout and 10 without trout) differing in chemical characteristics. Gut contents of trout, crayfish and eels were also examined to evaluate whether competition or predation could be linked to the patterns found. Abundances of crayfish and galaxiids were significantly lower in streams with trout, but in streams without trout, substrate size was one of the most important factors determining crayfish abundance. In contrast to crayfish, other macroinvertebrates were more abundant in trout streams than streams without trout and significantly more taxa were found in streams with trout. Macroinvertebrate abundance was related to environmental factors, such as pH, substrate, depth and total nitrogen. Gut content analysis showed an overlap in diet (mostly invertebrates) between trout and eels. Crayfish, however, had a more omnivorous diet where detritus was the most frequently occurring food material. Differences in chemical characteristics marked the streams with and without trout. Thus, trout were not present in streams with pH < 6.0. Crayfish and galaxiids were present in streams with pH ranging from 4.1 to 7.9, and those with pH < 6.0 may function as trout-free refuges where larger populations of these species may persist. However, other macroinvertebrate taxa may be more negatively affected by acidification than by trout. By protecting naturally acidic, brown water streams, New Zealand crayfish and galaxiid fish populations can be conserved within geographic areas where trout are present.     

Heterogeneous Atmospheric Nitrogen Deposition Effects Upon the Nitrate Concentration of Stream Waters in a Forested Mountain Area

Nitrogen compounds generated by anthropogenic combustion deposits in forest watersheds and induce nitrogen saturation of the area. Because excess nitrogen is derived from atmospheric deposition, this action is expected to uniformly affect a wide area of forest soils. Geographically, heterogeneous nitrate concentration of stream water within a small area has been attributed to the tree type, geological setting and tree cut. In this article, we hypothesized that the effect of the atmospheric nitrogen deposition in the forest watershed may vary within a small area, and that such variation is induced by the degree of air mass containing a high concentration of nitrogen deposition of combustion origin. We measured major ion concentrations, including nitrate, nitrite oxygen and nitrogen stable isotope of nitrate sampled at 24 water streams in the Chichibu region, which is 50?C100 km from the Tokyo metropolitan area. The nitrate concentration showed a wide range (25.6?C237 ??mol L^?1) within 300 km², which was explained sufficiently by the air mass advection path and its contact with the mountain??s surface. The nitrate concentration showed a significant positive correlation with chloride (r?=?0.73; p?<?0.001). As chloride originates outside of the Chichibu region, the positive correlation between two ions showed that the nitrate concentration of the stream water was affected by the nitrogen compound from the Tokyo Metropolitan area as a form of atmospheric deposition. Between the nitrate concentration and the stable isotope ratio of oxygen of nitrate, there was a positive correlation until nitrate concentration of 100 ??mol L^?1. When the nitrate is over 100 ??mol L^?1, ??¹⁸O shows a stable value of ca. 5.7??. This indicates that the nitrification proceeds when the nitrate concentration was low to middle, but the reaction slowed when the nitrate concentration became high. Oxygen stable isotope of nitrate along with a set of nitrate concentrations can be used as a good indicator of nitrogen saturation.     

Ecology of the native and introduced crayfishes Austropotamobius pallipes and Procambarus clarkii in southern Spain and implications for conservation of the native species

The former and present distribution of white clawed crayfish (Austropotamobius pallipes) in the province of Granada (southern Spain) is studied. Before 1980 it was widely distributed but at present only 16 populations exist. The decline is related to the presence of the freshwater red-swamp crayfish (Procambarus clarkii), an American species, vector of the aphanomycosis disease, introduced to the Iberian Peninsula in 1974 and now widely distributed in the watercourses and marshes of southern Spain. To establish an appropriate conservation policy for A. pallipes at its southernmost distribution limit, we studied watercourses from two river basins, Genil and Guadiana Menor, (tributaries of the Guadalquivir River). P. clarkii inhabits the medium to lower reaches of these two river basins (with its upper limit at 820 m a.s.l.). The distribution of this species was best explained by the effect of three of the 12 analyzed variables: altitude, water-current and minimum winter temperatures. From our results, the repopulation of the native crayfish is almost impossible in those reaches inhabited by P. clarkii. However, based on the habitat selection study, it is clear that upper reaches are unsuitable for the red-swamp crayfish, where the native white-clawed crayfish may have greater survival possibilities, and these sites can be used for future restocking projects.     

Selected ecosystem relationships in the mercury contaminated Wabigoon-English river system,Canada, and their underlying causes

Mercury concentrations in basic ecosystem compartments of the Wabigoon-English River system are interrelated. Statistically significant correlations are evident between Hg levels in various biotic-young northern pike (Esox lucius), yearling yellow perch (Perca flavescens) and crayfish (Orconected virihs) and abiotic compartments (water, surface sediments). Highly significant positive correlations are found between methyl-mercury in water and Hg in biota. Highly significant positive correlations also exist between Hg levels in different organisms, despite differences in habitat, diet, uptake routes and proportion of bodyburden that is in the methyl form. A plausible basis for these relationships is the rapid equilibration between methyl and inorganic Hg in and between various abiotic components of the ecosystem and subsequent incorporation into food webs. As sediment-water partitioning for inorganic and methylmercury is also rapid it is likely that in lakes and slow moving river reaches, local environmental variables probably largely govern Hg body burdens in biota.     

Environmental fate of mercury discharged into the upper Wisconsin river

We studied the distribution of Hg in sediments, fish, and crayfish in a 60 km reach of the Upper Wisconsin River that formerly received Hg in discharges from pulp and paper mills. The most heavily contaminated strata of sediments were deposited during the 1950s and early 1960s and buried under subsequent deposits; however, surficial sediments remained substantially enriched at certain sites in 1981. Median concentrations of Hg in surficial sediments, adjusted for grain size, were at least 10-fold greater at the main study area than at an upstream reference site. Total concentrations exceeded 1.0 μg g^?1 wet weight in axial muscle tissue in only 2 of 173 fish analyzed from the study area; however, historical comparisons revealed that Hg contamination of fish (common carp Cyprinus carpio and walleye Stizostedion vitreum vitreum) and crayfish (Orconectes) in the river had not decreased since the early 1970s. The availability of Hg to biota in this system may be enhanced by rapid methylation of the metal in surficial sediments, despite burial of the most heavily contaminated sediments. Management practices for this river should be designed to prevent conditions favoring mobilization and methylation of Hg in sediments.     

Relationship of biotic and abiotic factors to recruitment patterns in Margaritifera margaritifera

We investigated relationships of biotic and abiotic factors to recruitment patterns of the endangered freshwater pearl mussel (Margaritifera margaritifera) in 10 Swedish streams. We found that the maximum proportion of gravid mussels did not differ between streams with and without recent recruitment. Moreover, the mean glochidial load on trout (Salmo trutta), which was positively related to adult mussel density, did not differ significantly between these stream types. Thus, the larval stages of the freshwater pearl mussel were not related to recruitment failure. Instead, recruitment is probably hindered at the next stage in the life history of the mussels, the benthic stage, and may be related to sedimentation as turbidity was four times greater in streams lacking recent recruitment than in streams with recent recruitment. Furthermore, we found that juvenile mussel density was positively related to the number of glochidial infections per stream area in streams with ongoing recruitment, indicating that successful recruitment in these streams may depend on both mussel and trout density. Future research should thus examine biotic interactions between mussels and trout as well as the effects of sedimentation on benthic-living mussels.     

Simultaneous automated determination of chloride,nitrite, nitrate,and ammonia in water and wastewater

An automated system capable of simultaneous determination of chloride, nitrite, nitrate, and ammonia in about 2 ml of fresh water or wastewater is described. The four compounds are determined using modifications of established colorimetric procedures. Results can be reported at a true rate of five samples per hour with a relative standard deviation at optimum concentrations of less than 2%. Detection limits are 1 ppm Cl for chloride and 1, 5, and 5 ppb N for nitrite, nitrate, and ammonia, respectively. Sample pH adjustment is not required from 0.0002 N H₂SO₄ (pH 3.7) to 0.005 N NaOH (pH 11.2). Fresh water and wastewater samples were analyzed for nitrite and ammonia by the proposed procedure and by the manual sulfanilic-naphthylamine and nesslerization method. Analysis by the student t-test showed no significant difference between the paired sets of data (P > 0.5). When potable and wastewater samples were spiked with sodium chloride, potassium nitrate, sodium nitrite, and ammonium chloride and analyzed, average recoveries were 99 to 103%.     

The occurrence of primary burrowing crayfish in terrestrial habitat

A researcher’s perception of a target species’ landscape strongly influences the design of habitat studies conducted at broad spatial scales. Consequently, researcher-dependent perceptions may misguide conservation efforts. Although the life histories of some crayfish (i.e., primary burrowers) are centered on a fossorial existence independent of surface water, all North American crayfish are viewed in an aquatic context. This paradigm restricts the range of habitats that are typically sampled and managed for crayfish conservation. This study used presence/absence of the primary burrower Distocambarus crockeri at 137 locations within the Long Cane Ranger District of the Sumter National Forest, South Carolina, USA, to model the habitat association of the species across a GIS-based landscape. Logistic regression indicated that D. crockeri presence was most strongly associated with a terrestrial habitat defined by a set of morphologically similar soils located along ridge tops. Furthermore, the species was negatively associated with aquatic habitats such as streams and floodplains. The results indicate that D. crockeri is a terrestrial habitat specialist and should be modeled and managed at the landscape as a terrestrial organism. When viewed as a subset of the total United States cambarid fauna, primary burrowers are disproportionately imperiled. Primary burrowers comprise only 15% of the total crayfish fauna, while they account for 32% of those crayfish ranked critically imperiled. Habitat loss and an aquatic bias that restricted sampling to aquatic and semi-aquatic habitats might explain the group’s disproportionate imperilment.     

Assessing the Impact of Effluents from a Modern Wastewater Treatment Plant on Breakdown of Coarse Particulate Organic Matter and Benthic Macroinvertebrates in a Lowland River

Wastewater treatment plants (WWTP) with insufficient technologies for wastewater purification often cause a distinct nutrient pollution in the receiving streams. The increased concentrations of dissolved nutrients can severely disturb the ecological integrity of streams, which has been recently shown for basic ecosystem processes like mineralization of coarse particulate organic matter (CPOM). The present study investigated the impact of a modern WWTP (Zentralkläranlage Jena) on breakdown rates of CPOM exposed in net bags (1 mm mesh size) to the effluent of a large municipal WWTP and an upstream control site in the Saale River (Thuringia, Germany) from April to October 2005. Control and effluent site differed significantly in water chemistry with increased concentrations of dissolved organic carbon (DOC), ammonium, sulfate, and chloride at the effluent site, while the control site displayed higher concentrations of nitrate. However, breakdown rates of toothpickers and small twigs were not significantly different between the sites, whereas breakdown rate of leaf litter was significantly higher at the effluent site (k?=?0.0124 day^?1) than at the control site (k?=?0.0095 day^?1). Benthic invertebrate assemblages inhabiting the sandy stream bed at both sites were dominated by Chironomidae and Oligochaeta, typical inhabitants of fine sediments. Although the Shannon diversity of the benthic invertebrates was slightly higher at the effluent site (0.85) than at the control site (0.63), no significant difference could be detected. Bacterial numbers in water samples and surface biofilms on glass slides also displayed no significant differences between the two sites. This study showed that the effluents of a WWTP with modern technologies for wastewater purification did not directly affect breakdown rates of CPOM, bacteria numbers in epibenthic biofilms and the water column, and the community composition of sediment inhabiting aquatic macroinvertebrates in an effluent-receiving river with already increased concentrations of dissolved nutrients.     

Isolation and identification of iron-reducing bacteria from gley soils

Seventy-one facultative anaerobic bacteria, capable of reducing iron oxide in pure culture, were isolated from three differently gleyed subsoils. The bacteria were picked at random from poured plates (10⁻⁵ and 10⁻⁶) inoculated with serially diluted soil samples. An attempt was made to identify these strains by morphological and biochemical tests. Among these 71 iron-reducing bacteria, all except three were capable of reducing nitrate to nitrite and 35 reduced nitrite further into gaseous compounds (denitrification), but only one strain (Bacillus subtilis) produced H₂S. Based upon their physiological and morphological properties, 38 strains were allotted to the genus Pseudomonas, 31 sporeformers to the genus Bacillus and two were regarded to be coryneform (Arthrobacter?) bacteria. Species identified were Ps. denitrificans (23), ps. stutzeri (8) ps. fluorescens-putida (5), Bacillus cereus (6), B. cereus var. mycoides (14) and Bacillus subtilis (9). Two spore-forming bacilli, two non-pigmented pseudomonads and two coryneform type of bacteria could not be identified. The significance of the enzyme nitrate reductase (nitratase) of these bacteria for anaerobic respiration and as a mechanism of iron reduction is discussed.     

Effect of oils and oil-products on crustaceans

The effect of oil and oil products on the 5 species of crustaceans in the Caspian sea was investigated. Two species of shrimp (Palaemon elegans, P. adspersus), crab (Thithropanopeus harrisii tridentatus), amphipoda (Pontogammarus maeoticus), and cirripeda crayfish (Balanus improvisus) are classified among them. Various oil products (oil, phenol, fuel oil, xerosene, gasoline, solar oil) were used in the experiments. The influence of oil on the respiration and mass increase, quality of generation, and larval stages of the crustaceans was studied. Minimal critical concentrations for the crustaceans are 0.001 mg L^?1 of fenol, 0.4 mg L^?1 of oil, 0.1 mg L^?1 of solar oil, 0.01 mg L^?1 of gasoline and fuel oil.     

Dynamics of nitrification in a continuous flow system

Ammonium sulphate was added at constant rates to a column containing a mixture of glass beads and marble chips inoculated with the nitrifying bacteria, Nitrosomonas europaea and Nitrobacter agilis. Changing the rate of NH₄⁺ addition caused nitrite and nitrate concentrations in the outflow to stabilize at characteristic steady state values after brief transition periods. The steady state concentrations were inversely related to the flow rate. The transition phases were characterized by overshoots in the concentrations of nitrite and nitrate when the flow was reduced and a smooth monotonic change to the new steady state values when it was increased. Three mathematical models were developed based on several simplifying assumptions and tested with the data. The best fit was obtained when it was assumed that growth of the organisms could be described by a modification of the Verhulst-Pearl logistic equation.     

In Situ Batch Denitrification of Nitrate-Rich Groundwater Using Sawdust as a Carbon Source—Marydale, South Africa

Batch experiments were performed to denitrify groundwater using sawdust as a carbon source at Marydale, South Africa. Alkalinity, pH, electrical conductivity, nitrate, nitrite, ammonia, SO ₄ ^2? , heterotrophic plate count (HPC), dissolved organic carbon (DOC), potassium and chloride were monitored. Two soil depths, 75 to 100 and 165 to 200 cm, respectively, from the Marydale area were used as matrix material during denitrification based on contrasting chemical composition with respect to major ion composition and moisture to consider different denitrification rates for varying soil depths. Different N to C ratios were used to evaluate the denitrification efficiency and the least undesirable products, e.g., elevated SO ₄ ^2? , H₂S and other reduced compounds. DOC is directly proportional to the N to C ratio used. Nitrite was produced for most of the treatments as incomplete denitrification occurred. The incubation periods were 28 and 43 days, respectively. N to C ratios were 12.6:1, 24:1, 34:1 and 54:1. Longer incubation period and higher N to C ratio resulted in total removal of both nitrate and nitrite. The reaction was carbon-limited for lower N to C ratios. The denitrification rate was proportional to the carbon availability at any time during the experiment. There was no significant difference in denitrification using heterogeneous and homogeneous particle sizes for sawdust. Soil depth of 75–100 cm displayed a greater denitrification rate than 165–200-cm soil depth due to higher initial soil nitrate concentration. The method showed some specificity, as DOC, nitrite, nitrate, alkalinity and HPC were the only parameters that showed a change in concentration over the duration of the denitrification experiment under constant temperature and nitrogen gas atmosphere. DOC and HPC were unacceptable for domestic use, but methods such as boiling or chlorinating water can rid it of bacteria.     

Responses of soil nitrous oxide production and abundances and composition of associated microbial communities to nitrogen and water amendment

Soil moisture and nitrogen (N) are two important factors influencing N₂O emissions and the growth of microorganisms. Here, we carried out a microcosm experiment to evaluate effects of soil moisture level and N fertilizer type on N₂O emissions and abundances and composition of associated microbial communities in the two typical arable soils. The abundances and community composition of functional microbes involved in nitrification and denitrification were determined via quantitative PCR (qPCR) and terminal restriction length fragment polymorphism (T-RFLP), respectively. Results showed that N₂O production was higher at 90% water-filled pore (WFPS) than at 50% WFPS. The N₂O emissions in the two soils amended with ammonium were higher than those amended with nitrate, especially at relatively high moisture level. In both soils, increased soil moisture stimulated the growth of ammonia-oxidizing bacteria (AOB) and nitrite reducer (nirK). Ammonium fertilizer treatment increased the population size of AOB and nirK genes in the alluvial soil, while reduced the abundances of ammonia-oxidizing archaea (AOA) and denitrifiers (nirK and nosZ) in the red soil. Nitrate addition had a negative effect on AOA abundance in the red soil. Total N₂O emissions were positively correlated to AOB abundance, but not to other functional genes in the two soils. Changed soil moisture significantly affected AOA rather than AOB community composition in both soils. The way and extent of N fertilizers impacted on nitrifier and denitrifier community composition varied with N form and soil type. These results indicate that N₂O emissions and the succession of nitrifying and denitrifying communities are selectively affected by soil moisture and N fertilizer form in the two contrasting types of soil.     

Nitrogen isotope discrimination in denitrification of nitrate in soils

Nitrogen isotope discrimination during denitrification in soils of nitrate containing natural concentrations of ¹⁴N and ¹⁵N was studied by determining the amount and the ¹⁵N content of nitrate-N and (nitrate + nitrite)-N in nitrate-treated soils incubated under anaerobic conditions (He atmosphere) for various times after treatment with glucose to promote denitrification. Analyses performed showed that the nitrate-N lost on incubation of these soils could largely be accounted for as products of denitrification (nitrite, NO. N₂O and N₂).The studies reported show that marked discrimination between ¹⁴N and ¹⁵N occurs during denitrification of nitrate in soils and that significant N isotope effects occur both in reduction of nitrate to nitrite and in reduction of nitrite to gaseous forms of N. They also indicate that the overall N isotope effect during denitrification of nitrate in soil will depend upon the tendency of the soil to accumulate nitrite under conditions that induce denitrification.It is concluded that discrimination between ¹⁴N and ¹⁵N during denitrification in soils of nitrate containing natural concentrations of these isotopes is of sufficient magnitude to invalidate the use of N isotope-ratio analyses for assessment of the contributions of soil and fertilizer N to nitrate in surface or ground waters or to nitrous oxide in the atmosphere.     

Effects of liming on crayfish and fish in Sweden

The effects of lime treatment on crayfish (Astacus astacus and Pacifastacus leniusculus) populations in 17 lakes and fish populations in 47 lakes and 7 rivers within the trial period 1976–82 have been evaluated. An increase in the catch of crayfish per unit effort was observed in 7 lakes, although significantly in one lake only. The varying results in the other lakes indicate that factors other than pH may be of greater importance for the development of crayfish populations after liming. Recruitment of fish improved in waters where liming resulted in pH >5.5. In lakes with pH <5.5 before and pH >5.5 after treatment, there was a significant increase in the number of fish caught, from 12 to 34 per unit effort. Due to improved recruitment the individual average weight was smaller and hence the catch in weight per unit effort was about the same as before liming. After lime treatment in streams which resulted in a stable pH of >5.5, the abundance of juvenile salmonids increased to numbers found in non-acidified streams. In other streams acid spates reduced the positive influence of liming on the abundance of juvenile salmonids.     

Effect of Water Bicarbonate Concentration,pH and the Presence,or Not,of a Nitrification Inhibitor in the Nitrification Process

Ammonium (NH₄⁺) is an important nitrogen (N) source for plant growth. Nevertheless, NH₄⁺–N can be oxidized to nitrate (NO₃^?) by nitrification and then nitrate and nitrite can be leached into groundwater. The aims of this study were to investigate the effect of the water bicarbonate concentration, pH, and the presence, or not, of a nitrification inhibitor, on the nitrification process. Six treatments were established, changing the pH and the bicarbonate concentration, with or without the nitrification inhibitor. The results showed that an active nitrification process occurred in treatments with high pHs (8 and 7) and with no nitrification inhibitors. This causes an increase in the nitrate and nitrite concentration in the substrate. The use of N–NH₄⁺ fertilizers joint to a nitrification inhibitor, especially in nutritive solutions with a high concentration of bicarbonate and a pH of 8 and 7, decreases nitrate and nitrite accumulation in substrate which can prevent nitrate and nitrite leaching.     

Sources of acidity in running waters in central northern Sweden

Factors controlling the acidity of running waters between the coast of the Gulf of Bothnia and the Caledonian mountain range in central-northern Sweden were studied intensively in 8 large streams and in two synoptic surveys of 179 small streams. The bulk deposition of SO₄ ^2? was between 11–22 μeq m^?2 y^?1, of which 93% was nonmarine, with the highest values in the coastal region. Organic anions were the most frequent acid anions in the whole investigation area followed by sulfate. The major portion of SO₄ ^2? was from natural sources in the whole investigation area. The most acidic streams occurred in the hilly wave-washed terrain of the coastal region, because of a high terrestrial export of organic acids and very low neutralizing capacity. It is concluded that most of the acidity in the investigated streams is due to natural sources.     

Contrasting response of two forest soils to nitrogen input: rapidly altered NO and N2O emissions and nirK abundance

Denitrification represents one of the main microbial processes producing the primary and secondary greenhouse gases nitrous oxide (N₂O) and nitric oxide (NO) in soils. It is well established that abiotic factors like the soil water content and the availability of nitrogen (N) are key parameters determining the activity of denitrifiers in soils. However, soils differing regarding their characteristics such as the content of C_org, the soil texture or the pH value may respond in specific manners to equivalent changes in soil moisture and N input. Thus, short-term incubation experiments were performed to test and compare the capacity of two contrasting Austrian forest soils to respond to mineral N application at increased soil water contents. Soils from the pristine Rothwald forest (rich in C_org) and the more acidic Schottenwald forest (poor in C_org) were amended with either NH ₄ ⁺ -N or NO ₃ ^? -N and were incubated at 40% and 70% water-filled pore space for 4 days. Changes in mineral N pools, nitrite reductase activity and NO and N₂O emission rates were measured, and the abundance and structural community composition of the functional group involved in nitrite reduction were analysed via quantitative real-time polymerase chain reaction and terminal restriction fragment length polymorphism analysis of the nirK gene. Rapid and distinct activity responses to increased soil moisture and altered mineral nitrogen availability were observed in two contrasting forest soils. In both soils, nitrogen oxide emission rates were stimulated by N inputs and, depending on the soil moisture status, either NO or N₂O emission was prevailing. However, different N cycling processes appeared to predominate in either soil under equivalent treatment. Nitrogen oxide emissions peaked following NO ₃ ^? application in Schottenwald soils but were the highest after NH ₄ ⁺ application in Rothwald soils. Denitrifying (nirK) communities differed significantly in Rothwald and Schottenwald soils; however, changes in the community structure were marginal during the short-term incubation. Abundances of nirK genes remained unaffected by N application in either soil. The soil water content affected nirK gene abundances only in Rothwald soil, indicating a distinct reaction of nitrite reducing communities in the two soils.