Environmental transmission of a sulfur-oxidizing bacterial gill endosymbiont in the tropical lucinid bivalve Codakia orbicularis.

Codakia orbicularis is a large tropical member of the bivalve mollusk family Lucinidae which inhabits shallow-water sea-grass beds (Thalassia testudinum environment) and harbors sulfur-oxidixing endosymbiotic bacteria within bacteriocytes of its gill filaments. When a C. orbicularis-specific 16S rDNA (DNA encoding rRNA) primer is used with a bacterium-specific 16S rDNA reverse primer in amplifications by PCR, the primer set was unsuccessful in amplifying symbiont DNA targets from ovaries, eggs, veligers, and metamorphosed juveniles (600 microns to 1 mm in shell length) cultivated in sterile sand, whereas successful amplifications were obtained from gill tissue of adult specimens and from metamorphosed juveniles (600 microns to 1 mm in shell length) cultivated in unsterilized sea-grass bed sand. To ascertain the presence of the symbiont target in juveniles, restriction fragment length polymorphism analysis, Southern blotting, and transmission electron microscopy were used. Specific hybridizations and observation of endosymbiotic bacteria in the gills of numerous juveniles cultivated in unsterilized sea-grass bed sand showed that the sulfur-oxidizing endosymbionts of C. orbicularis are environmentally transmitted to the new generation after larval metamorphosis.     

Putative environmental transmission of sulfur-oxidizing bacterial symbionts in tropical lucinid bivalves inhabiting various environments

Four tropical lucinids, Codakia orbiculata, C. pectinella, Linga pensylvanica, which inhabit sea-grass beds, and Lucina pectinata, which inhabits mangrove swamps, harbor sulfur-oxidizing endosymbiotic bacteria within bacteriocytes of their gill filaments. To elucidate the symbiont transmission mode in these bivalves, symbiont-specific oligonucleotides were designed and used in polymerase chain reaction amplifications (PCR). For all species investigated, each primer set was unsuccessful in amplifying symbiont DNA targets from ovaries and testis, whereas successful amplifications were obtained from symbiont-containing gill tissue. These data suggest that the transmission mode is environmental, independently of the lucinid habitat, as it is in the other tropical lucinid Codakia orbicularis.     

Gill filament differentiation and experimental colonization by symbiotic bacteria in aposymbiotic juveniles of Codakia orbicularis (Bivalvia: Lucinidae)

Summary

Codakia orbicularis is a tropical lucinid harboring gill endosymbionts which are environmentally transmitted from a free living-symbiont form to the new host generation after metamorphosis. Structural changes occurring in the cellular organization from incomplete gill filaments in young aposymbiotic juveniles to full differentiated gill filaments containing bacterial endosymbionts in reared symbiotic juveniles, were analyzed for juveniles from 250 μm to 2 μm shell-length. Aposymbiotic juveniles possess differentiated gill filaments with ciliated, intermediary, and lateral zones similar to those described in wild juveniles, except for the bacteriocytes which are lacking. Granule cells, which progressively differentiate during the morphogenesis of the gill filament, do not appear as a consequence of symbiosis. Experimental colonization of aposymbiotic juveniles by the free-living symbiont form has been obtained through the addition of unsterilized sand collected from the natural habitat of C. orbicularis. Two days after exposure to crude sand, symbiosis-competent bacteria enter by endocytosis at the apical pole of undifferentiated cells which progressively differentiate into classical bacteriocytes similar to those found in the adult gill filaments. Undifferentiated cells of aposymbiotic gill filaments remain receptive to bacteria several months after metamorphosis, and become bacteriocytes when aposymbiotic juveniles get contact with the symbiont free-living form. Therefore, the environmental transmission of symbionts does not appear to be restrained to a defined period of time during post-larval development in C. orbicularis.     

The influence of temperature on the susceptibility of vine weevil, Otiorhynchus sulcatus (Fabricius) (Coleoptera: Curculionidae), larvae to Metarhizium anisopliae (Deuteromycotina: Hyphomycetes)

Mortality of Otiorhynchus sulcatus (Fabricius) larvae at 10^oC, 15^oC, 20^oC and 25^oC following treatment with 10⁷conidia ml“¹suspensions of six Metarhizium anisopliae (Metschnikoff) Sorokin isolates was temperature dependent. In all cases, the LT₅₀s were inversely related to temperature, but the nature of this response varied between isolates. Strain 101-82 was the most virulent isolate at 25^oC with an LT₅₀ of 3.7 days, but it was the least virulent isolate at 15^oC and it failed to kill any O. sulcatus larvae at 10^oC. In contrast, strain 159-83 had the lowest virulence at both 20^oC and 25^oC, whereas it was the most virulent isolate at 10^oC with an LT₅₀ of 20.0 days. The mortality rates followed a similar pattern and were positively related to temperature in all cases with the exception of strain 159-83 at 25^oC. Mycosis development was examined on larvae treated with strain 275-86 and significant differences were obtained between all four temperatures. Sporulation commenced after approximately 2.75 days at 25^oC, but took nearly 11 days at 10^oC. The infection rates also varied between temperatures; sporulation occurred on 98% of the treated larvae at 25^oC, but only on 93%, 87% and 49% of the larvae at 20^oC, 15^oC and 10^oC, respectively. The results of these bioassays demonstrate that temperature has a significant effect on the virulence of M. anisopliae. The differences between fungal strains also emphasises the importance of selecting isolates for specific situations on the basis of their temperature profiles.     

Larval development of the invasive charru mussel,Mytella strigata (Bivalvia: Mytilidae)

ABSTRACT

The South American charru mussel, Mytella strigata, was recently recorded in Singapore waters, possibly introduced into Southeast Asia through shipping. The mussels have rapidly spread across estuarine coastal mudflats. Adult mussels were collected, spawned in aquaria and larvae were successfully cultured to the juvenile stage in the laboratory. The larval morphology and development of M. strigata is described in this paper. D-shaped veligers were produced within 20 h of fertilization and were approximately 75 µm in shell length. These larvae were capable of settlement two weeks post fertilization. Given an adequate amount of food, they were able to grow up to 1 mm in shell length within 30 days. The larval shell of M. strigata possesses anterodorsal G2 hinge teeth as distinct wavy ledges, with a pitted resilial ridge clearly evident in the juvenile shell.     

Modelling the Effects of Water Stress and Temperature on Seed Germination of Radish and Cantaloupe

Cantaloupe (Cucumis melo L.) and radish (Raphanus sativus L.) are considered as important vegetables with potential for national and international markets due to their sugars, vitamins and minerals. This study arranged, therefore, to simultaneously investigate the effect of temperature (T) and water potential (ψ) on seed germination (SG) of these plants using two hydrothermal time (HTT) models and to determine cardinal Ts and base water potential (ψ_b(50)) for both species. The results indicated that SG of both species was more affected by ψ than T (p ≤ 0.001). At Ts below an optimum temperature (T_o) the ψ_b(50) was constant (− 0.582 and − 0.760 MPa for radish and cantaloupe, respectively) and then increased linearly by 0.0481 and 0.0446 MPa °C⁻¹ as T increased above T_o (as thermoinhibition) until 0 MPa at the ceiling temperature (T_c), respectively. As the first report, however, we observed that the T at which ψ_b(50) begins to change was the same here (that is, T_d = T_o), when determined by either model for both species. This result suggests that the assumption in Rowse and Finch-Savage’s model (T_d is often less and or very close to T_o) may be invalid in some cases. For both species, the base temperature (T_b) and T_o were not affected by ψ and were constant while there was an exception only for T_c for which the value declined with decreasing ψs (more negative). In general, the estimated T_b, T_o and T_c were 9.64, 21.3 and 33.0 °C for radish and 11.8, 28.3 and 45.7 °C for cantaloupe in the control condition (ψ = 0 MPa), respectively. The HTT models used here and their parameters, each with strengths and weaknesses, can be used as a predictive tool in both cantaloupe and radish SG simulation models. However, at first, we need to select an appropriate HTT model based on SG behavior of plant species and then use the best model for quantifying the response of SG across Ts and ψs.

     

Bioefficacy and mode of action of rocaglamide from Aglaia elaeagnoidea (syn. A. roxburghiana) against gram pod borer,Helicoverpa armigera (Hübner)

Abstract: Rocaglamide, a highly substituted benzofuran, was isolated and identified as the main biologically active component in Aglaia elaeagnoidea (syn. A. roxburghiana) for gram pod borer Helicoverpa armigera (Hübner). Addition of rocaglamide to an artificial diet retarded the growth of neonate larvae in a dose‐dependent manner with EC₅₀ values of 0.76 p.p.m. These values compared favourably with azadirachtin (EC₅₀ = 0.23 p.p.m.). However, azadirachtin was apparently more potent than rocaglamide in inducing growth inhibition via oral administration to these first stadium larvae. The candidate compound was found to have LD₅₀ and LD₉₅ values of 0.40 and 1.02 μg per larva, respectively, in topical application against third instar larvae 96 h post‐treatment. However, these values for azadirachtin were 8.16 and 25.8 μg per larva for the same period. This shows that azadirachtin was less effective against third instar H. armigera larvae in inducing acute toxicity via topical treatment in comparison with rocaglamide. However, severe morphological larval deformities were observed in such azadirachtin‐treated larvae during the process of ecdysis. The cytotoxic nature of rocaglamide was established by evaluating dietary utilization and the results did not implicate any antifeedant effect but the toxicity‐mediated effect due to reduced efficiency of conversion of ingested food. It was obvious that feeding deterrence is not the primary mode of action but a centrally mediated effect, which could be due to the induced cytotoxicity at non‐specific cellular levels.     

ON THE EXISTENCE OF A Go-PHASE IN THE CELL CYCLE

The model is based on the assumption that the cell cycle contains a G_o-phase which cells leave randomly with a constant probability per unit time, γ. After leaving the G_o-phase, the cells enter the C-phase which ends with cell division. The C-phase and its constituent phases, the‘true’G₁-phase, the S-phase, the G₂-phase and mitosis are assumed to have constant durations of T, T₁T_s, T₂ and T_m, respectively. For renewal tissue it is assumed that the probability per unit time of being lost from the population is a constant for all cells irrespective of their position in the cycle. The labelled mitosis curve and labelling index for continuous labelling are derived in terms of γ, T, and T_s. The model generates labelled mitosis curves which damp quickly and reach a constant value of twice the initial labelling index, if the mean duration of the G_o-phase is sufficiently long. It is shown that the predicted labelled mitosis and continuous labelling curves agree reasonably well with the experimental curves for the hamster cheek pouch if T has a value of about 60 hr. Data are presented for the rat dorsal epidermis which support the assumption that there is a constant probability per unit time of a cell being released from the Go-phase.     

Steroid hormones promote bovine oocyte growth and connection with granulosa cells

Many approaches have been investigated for growing oocytes in vitro in mammals. To support oocyte growth in vitro, the culture systems must meet certain conditions for maintaining connections between oocytes and surrounding granulosa cells. The aims of this study were to determine the effects of combinations of 17β-estradiol (E₂) and androstenedione (A₄) on in vitro growth of bovine oocytes and to determine the number of connections between the oocyte and granulosa cells. Oocyte–granulosa cell complexes (OGCs) collected from early antral follicles (0.4−0.7 mm in diameter) were cultured for 14 days in a medium with different concentrations of E₂ and A₄, either alone or in combinations. We then assessed the number of transzonal projections (TZPs), which extend from granulosa cells through the zona pellucida to the oolemma. During in vitro growth culture, OGC structures were maintained in the medium with steroid hormones. The mean diameter of oocytes grown in the medium with both E₂ and A₄ was increased from 95.8 μm to around 120 μm, larger than oocytes grown without steroid hormones (109.9 μm) and similar in size to in vivo fully grown oocytes (119.4 μm) from 4- to 6-mm antral follicles. In subsequent in vitro maturation culture (22 hours), 30% (12 of 40) and 34% (14 of 41) of oocytes grown with E₂ or A₄ alone, respectively, matured to metaphase II; meanwhile, oocytes grown with a combination of E₂ and A₄ matured to metaphase II at a high rate (58%, 23 of 40). Growing oocytes isolated from early antral follicles had many uniformly distributed TZPs throughout the zona pellucida. After 14 days of culture, there was a significant decrease in the number of TZPs in oocytes grown without steroid hormones, whereas the number of TZPs was maintained in oocytes grown with steroid hormones. In particular, oocytes grown with E₂ alone or with a combination of E₂ and A₄ had numbers of TZPs similar to oocytes before growth culture. In conclusion, a combination of E₂ and A₄ maintained the connections between oocytes and granulosa cells during in vitro growth culture of bovine oocytes for 14 days, resulting in the complete oocyte growth and the acquisition of meiotic competence in more than half the oocytes.     

Effect of temperature on development rate and intrinsic rate of increase ofAphis gossypii reared on greenhouse cucumbers

Thermal requirements for development and life table statistics ofAphis gossypii Glover (Homoptera, Aphididae) were determined over a range of constant temperatures from 10 to 30°C. The lower development threshold and the sum of effective temperatures were 6.9°C and 90.1°C, respectively, for preimaginal development, and 5.8°C and 113.6°C from birth to the onset of reproduction. Mean total fecundity ranged from 36 larvae per female at 10°C to 76 larvae at 30°C. On a time scale of days, net reproductive rate (R _o ) increased with increasing temperature while generation time (T) decreased causing the intrinsic rate of increase (r _m ) to increase linearly from 0.115 to 0.465. On a day-degree scaler _m only varied from 0.019 to 0.028 because the growth ofR _o was compensated by an increase inT with increase in temperature. The nearly constantr _m in terms of day-degrees, over a wide range of temperatures, greatly simplifies the prediction of future population numbers ofA. gossypii.     

Veligers of an introduced bivalve,Limnoperna fortunei,are a new food resource that enhances growth of larval fish in the Paraná River (South America)

1. Larvae of ‘sábalo’, Prochilodus lineatus, whose adults represent over 60% of overall fish biomass in the Río de la Plata Catchment, have been observed to feed intensively on veligers of the exotic bivalve Limnoperna fortunei. 2. To assess the effects of this dietary shift on the growth of P. lineatus, 28‐day laboratory experiments were carried out feeding newly hatched P. lineatus larvae with three diets: zooplankton artificially enriched with L. fortunei veligers; natural zooplankton; and zooplankton artificially enriched with cladocerans and copepods. The average length, weight and gut contents of the fish larvae were assessed weekly and metabolic rates of fish larvae were measured. 3. Proportions of veligers in gut contents were always higher than those in the experimental diet: 100, 76 and 21% for veliger‐enriched, natural and low‐veliger diets, respectively. Larvae fed a veliger‐enriched diet grew to a significantly larger size than larvae fed the other two diets. In energetic balance comparisons using metabolic rates and prey energy content, all three diets were sufficient to support metabolism and growth. The greatest values of excess energy at the end of each week were in the veliger‐enriched experiments. 4. Feeding on veligers of L. fortunei significantly enhances the growth of P. lineatus larvae and supports the idea that this new and abundant resource is selectively preyed upon by P. lineatus during its larval stage. Higher growth rates may stem from the higher energy contents of veligers compared to crustaceans and/or from the lower energy costs of capturing slower prey.     

The effects of gradual or abrupt changes in salinity on digestive enzymes activity of Caspian kutum,Rutilus kutum (Kamensky, 1901) larvae

This study investigates the effects of gradual or abrupt changes in rearing salinity on food transit time and digestive enzymes activity of Caspian kutum (Rutilus kutum) larvae. The larvae (532 ± 0.05 mg) were supplied and randomly allocated into 12 tanks at a density of 45 fish per tank. Experimental treatments were fresh water (salinity 0) [FW] as control, exposure to salinity 5 [T₁], and gradual transfer to salinity 10 in two steps of first to 5 h, then and after 12 h to a salinity of 10 [T₂], and abrupt change (direct transfer to a salinity of 10 [T₃]). Results showed at 8 h after start of feeding that the larvae intestine was filled with food pellets except in treatment T₁. Enzyme activity responded to salinity change as follows: the highest trypsin, amylase, and chymotrypsin activities were observed in T₁; however, these were not significantly different to treatment T₃ (P > 0.05). Trypsin activity peaks in the FW and T₂ groups occurred 8 h after feeding, and in T₃ and T₁ groups 5 h after feeding. Peak chymotrypsin and alkaline phosphatase activity was observed 5 and 8 h after feeding in all experiments, respectively. The highest α amylase activity in FW and T₂ groups occurred 5 h after feeding, while in T₃ and T₁ these peaks were observed 8 h after feeding. These results indicate that salinity had some noticeable effects on the activities of digestive enzymes after feeding.     

Larvae of the invasive species <Emphasis Type="Italic">Limnoperna fortunei</Emphasis> (Bivalvia) in the diet of fish larvae in the Paraná River,Argentina

To assess the trophic impact of the planktonic larvae of the invasive Asian bivalve Limnoperna fortunei, introduced in South America around 1990, we investigated the gut contents of fish larvae collected at monthly intervals between October 2000 and March 2001 at three locations along a 600 km stretch of the Paraná River, and during November 2004 in two areas of marginal lagoons connected to the river near the city of San Nicolás, Argentina. Zooplankton was also collected in the lagoons in 2004. In total, 11,956 fish larvae were retrieved, of which 1,511 were used for detailed analyses. Of the 15 fish taxa collected, 11 had veligers of L. fortunei in their gut. Fish larvae with empty guts represented 60% (San Nicolás) to 72% (Paraná River) of the total number of fish. Proportions of feeding fish larvae with L. fortunei veligers in their guts varied between 20% (San Nicolás) and 56% (Paraná River); in 15% of the guts analyzed, L. fortunei was the only food item recorded. For those specimens that had consumed L. fortunei larvae and any other food item, L. fortunei was the most important item in 55% (Paraná River) to 71% (San Nicolás) of the animals in terms of biomass. No major temporal or spatial changes in the diet were observed along the Paraná River, but the relative biomass contribution of L. fortunei larvae differed strongly in fishes of different developmental stage. In protolarvae and mesolarvae, veligers accounted for 30–35% of the gut contents. In metalarvae, veligers accounted for only 3%, indicating enhanced food supply for the earliest fish life stages. Comparison of the relative proportions of the three main zooplankton types (L. fortunei veligers, cladocerans, and copepods) in the water and in larval fish guts indicates that L. fortunei is always selected positively over the other two prey types. While our results strongly suggest that the expansion of L. fortunei results in an enhanced food supply for local fish populations, they do not necessarily imply that the overall effect on the ecosystem in general, and on the fish fauna in particular is beneficial.     

A hydrothermal time model explains the cardinal temperatures for seed germination

Temperature (T) and water potential (y) are two primary environmental regulators of seed germination. Seeds exhibit a base or minimum T for germination (T_b), an optimum T at which germination is most rapid (T_o), and a maximum or ceiling T at which germination is prevented (T_c). Germination at suboptimal T can be characterized on the basis of thermal time, or the T in excess of T_b multiplied by the time to a given germination percentage (t_g). Similarly, germination at reduced y can be characterized on a hydrotime basis, or t_g multiplied by the y in excess of a base or threshold y that just prevents germination (y_b). Within a seed population, the variation in thermal times to germination among different seed fractions (g) is based on a normal distribution of y_b values among seeds (y_b(g)). Germination responses across a range of suboptimal T and y can be described by a general hydrothermal time model that combines the T and y components, but this model does not account for the decrease in germination rates and percentages when T exceeds T_o. We report here that supra‐optimal temperatures shift the ψ_b(g) distribution of a potato (Solanum tuberosum L.) seed population to more positive values, explaining why both germination rates and percentages are reduced as T increases above T_o. A modified hydrothermal time model incorporating changes in ψ_b(g) at T > T_o describes germination timing and percentage across all T and ψ at which germination can occur and provides physiologically relevant indices of seed behaviour.     

Daily Rhythms of Metabolic Rate and Body Temperature of Two Murids from Extremely Different Habitats

Daily circadian rhythms of body temperature (T_b) and oxygen consumption (VO₂) were measured in two murid species, which occupy extremely different habitats in Israel. The golden spiny mouse (Acomys mssalus) is a diurnal murid distributed in arid and hot parts of the great Syrio-African Rift Valley, while the broad-toothed field mouse (Apodeinns mystacinus) is a nocturnal species that inhabits the Mediterranean woodlands. In both species, the daily rhythms of T_b and VO₂ are entrained by the photoperiod. Under laboratory experimental conditions (ambient temperature T_a = 33^oC and photoperiod regime of 12L: 12D), Acomys russatus exhibits a tendency towards a nocturnal activity pattern, compared to the diurnal activity displayed by this species under natural conditions. Under the same photoperiod regime and at T_a = 28^oC, Apodemus mystacinus displays nocturnal activity, as observed under natural conditions. The maximal values of T_b were recorded in Acomys russatus at midnight (23:50 h), while the maximal values of VO₂ were recorded at the beginning of the dark period (18:20 h). In Apodemus mystacinus, the maximal values of T_b and VO₂ were recorded at 23:40 and 20:00 h, respectively. The ecophysiological significance of these results is discussed further.     

Gonadogenesis in early developmental stages of Acipenser naccarii and influence of estrogen immersion on feminization

Gonad development processes and the effects of a single 8‐hour immersion treatment with 17β‐estradiol (E2, 400 μg L⁻¹) on sex differentation in the Adriatic sturgeon, Acipenser naccarii, were investigated. After migration of germ cells, gonadal ridges appeared in 16‐ to 18‐day old larvae and undifferentiated gonads in 55‐ to 60‐day old larvae. Putative ovaries with notches in the germinal epithelium and presumed testes with smooth germinal epithelium appeared in 180–185‐day old juveniles. Ovaries with proliferating oogonia and early meiotic oocytes clusters were observed in 292‐day old juveniles. Testes did not exhibit germ cell mitosis until 430 days of age. Developmental stages in E₂‐treated animals closely followed those of controls up to 430 days. The treatment significantly increased the percentage of ovaries when administered to embryos about 1.5 day before hatching, while did not significantly altered the normal 1/1 sex ratio when administered to 1.5‐day old pre‐larvae and 10‐day old larvae. It is likely that in A. naccarii exogenous E₂ administration may act through a feedback mechanism of self‐supporting steroid production and that steroids are the physiological inducers of sex differentiation, as in most teleosts. The E₂‐immersion treatment, easier than time‐consuming administration through food, could be a good approach to control sex differentiation and caviar production.     

Larvicidal activity of some fungal extracts on Aedes caspius and Culex pipiens (Diptera: Culicidae)

A preliminary study was conducted to evaluate the effects of nine methanol fungal extracts on fourth instar larvae of Aedes caspius and Culex pipiens (Diptera: Culicidae). None of the extracts tested showed larvicidal activity except for the Paecilomyces lilacinus extract, which showed 100 % mortality against the larvae of Ae caspius after 24 h. The LC₅₀ values of the P. lilacinus extract after 24 h were 190.66 μg/mL against Ae. caspius and 254.25 μg/mL against Cx. pipiens, respectively. After 48 h, meanwhile, the LC₅₀ values were 65.70 and 164.13 μg/mL, respectively. Histological analysis of the midgut of the treated larvae of Ae. caspius and Cx. pipiens revealed changes such as cell destruction, spacing between the cells and disruption of the microvilli, resulting in an appearance of vacuolization in the midgut. This article is the first report of the use of P. lilacinus extract for the control of Ae. caspius and Cx. pipiens larvae and the data obtained may help to provide a better understanding of the mode of action of P. lilacinus as an insecticide against mosquito larvae.     

Effects of copper on production of periphyton, nitrogen fixation and processing of leaf litter in a Sierra Nevada, California, stream

SUMMARY.

• 1 Production of periphyton, nitrogen fixation and processing of leaf litter were examined in an oligotrophic Sierra Nevada stream and the responses of these processes to copper (2.5, 5 and 10μg 1^-1 Cu_T [total filtrable copper]; approximately 12, 25 and 50 ng 1^-1 Cu²⁺) were determined.
• 2 Autotrophic and total production were estimated from 3-week accumulations of biomass on artificial substrates. Mean autotrophic production in the control ranged from 0.22 to 0.58 mg C m^-2 h^-1 in summer-autumn 1979, but declined to 0.08–0.28 mg C m ² h^-1 after peak discharge in summer 1980, apparently due to phosphorus-limited growth. Total production in the control ranged from 0.30 to 0.82 mg C m^-2 h ^-1 in summer-autumn 1979 and from 0.16 to 0,68 mg C m ^-2 h ^-1 in 1980. Mean autotrophic productivity, estimated by ^l4C-bicarbonate uptake in daylight, ranged from 0.30 to 2.8 mg C m^-2 h^-1.
• 3 Autotrophic productivity was reduced by 57–81% at 2.5μg 1^-1 Cu_T, 55–96% at 5μg 1^-1 Cu_T, and 81–100% at 10μg 1^-1 CU_T, Heterotrophic productivity (based on dark ³⁵S-sulphate uptake) was inhibited to a lesser extent (28–63% at 2.5μg 1^-1 Cu_T, 24–84% at 5μg 1^-1 Cu_T, and 67–92% at 10μg 1^-1 Cu_T), The inhibition of autotrophic and heterotrophic productivity persisted through the year of exposure. Production in stream sections previously exposed to 2.5 and 5μg 1^-1Cu_T increased to control levels within 4 weeks after dosing, but remained depressed for more than 7 weeks after exposure to 10μg 1^-1 Cu_T.
• 4 The specific rate of photosynthesis (mg C mg chlorophyll a^-1 h^-1) of mature periphyton communities declined at all test concentrations of copper, but the rate for periphyton on newly-colonized surfaces did not change. The species composition of benthic algae shifted during exposure to an assemblage more tolerant of copper. Achrtanthes minutissima and Fragilaria crotonensis were the primary replacement species on newly-colonized surfaces.
• 5 The nitrogenase activity of blue-green algae was low. with controls ranging from 2.4 to 12 nmol C₂H₂ m^-2 h^-1. Nitrogenase activity was inhibited during the initial weeks of exposure by 5 and 10μg 1^-1 Cu_T. However, after 9 months of exposure, control and copper-treated sections did not differ.
• 6 The rate of processing of leaf litter, estimated by microbial respiration and nutrient quality of litter of resident riparian woodland taxa, was inhibited at all test concentrations of copper.