Effects of ammonia on growth,digestion and antioxidant capacity in juvenile yellow catfish Pelteobagrus fulvidraco (Richardson, 1846)

Effects of ammonia stress on food ingestion, growth, digestion and antioxidant capacity were investigated in juvenile yellow catfish Pelteobagrus fulvidraco (Richardson) with initial body weights of 20.24 ± 0.18 g. The fish were reared in triplicate in 15 experimental tanks at a rate of 30 fish per tank for 56 days. Water was maintained at a dissolved oxygen (DO) level of over 6.2 mg L^?1, pH 7.2–7.6, and temperature of 29.0 ± 1.5°C under a natural 12L: 12D photoperiod. Survival, food ingestion (FI), specific growth rate (SGR), food conversion efficiency (FCE), apparent digestibility coefficient (ADC), total antioxidant capacity (T‐AOC), levels of glutathione (GSH) and malonaldehyde (MDA), and activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH‐PX) of the juveniles were determined in total ammonia nitrogen (NH₃‐N+NH₄‐N) levels of 0 (control group), 3.36, 6.72, 13.44 and 26.88 mg L^?1. The results show that the survival, FI, SGR, FCE, and ADC decreased significantly with an increase in total ammonia nitrogen (p < .05), and a significantly negative correlation between SGR and total ammonia nitrogen levels (p < .05). T‐AOC, SOD, CAT, GSH‐PX and GSH in the blood, liver and gills were found to decline significantly with an increase in the total ammonia nitrogen level (p < .05), while the MDA in the blood, liver and gills was elevated significantly with the increase in total ammonia nitrogen (p < .05). The results indicate a threshold in the induction of the T‐AOC and activities of antioxidant enzymes in yellow catfish tissues with a total ammonia nitrogen increase. In the present study the total ammonia nitrogen threshold thus changed from 6.72 mg L^?1 in the juvenile yellow catfish.     

Seasonal variation of cyanobacteria and microcystins in the Nui Coc Reservoir, Northern Vietnam

In order to understand the environmental variables which promote the proliferation of cyanobacteria and variation in microcystin concentrations in the Nui Coc reservoir, Vietnam, physicochemical parameters, the occurrence, and abundance of phytoplankton, cyanobacteria, and microcystin concentration were monitored monthly through the year 2009–2010. The relationships between these parameters were explored using principal component analysis (PCA) and Pearson correlation analysis. The phytoplankton community was mainly dominated by the cyanobacterium Microcystis with higher cyanobacteria abundance during summer and autumn season. PCA and Pearson correlation results showed that water temperature and phosphate concentration were the most important variables accounting for cyanobacteria, Microcystis, and microcystin occurrence. Analysis of the toxins by high-performance liquid chromatography demonstrated the presence of two microcystin variants: microcystin-LR (MC-RR) and microcystin-ddRR (MC-ddRR) with total concentrations of the toxins in filtered samples from surface water ranging from 0.11 to 1.52 μg MC-LR equiv L^?1. The high concentrations of microcystin in the Nui Coc reservoir highlighted the potential risk for human health in the basin. Our study underlined the need for regular monitoring of cyanobacteria and toxins in lakes and reservoirs, which are used for drinking water supplies, not only in Vietnam but also in tropical countries.     

Interaction of larval age and antibiotic on the susceptibility of three insect species to Bacillus thuringiensis

The addition of chlortetracycline hydrochloride to bioassay diets increased the LC₅₀S of the HD-1 isolate of Bacillus thuringiensis 2.4–67.1 times when bioassayed against 4-day-old larvae of Trichoplusia ni, Heliothis virescens, and Ostrinia nubilalis. There were no significant differences in LC₅₀s when bioassays were conducted with neonate larvae in the presence or absence of antibiotic, nor were there any highly significant differences between the LC₅₀s for HD-1 when neonate larvae were used in the presence or absence of antibiotic or when 4-day-old larvae were used in the absence of antibiotic. Viable counts of B. thuringiensis did not increase over time in any of three bioassay diets. There were no differences in activity of pure chlortetracycline hydrochloride and veterinary grade Aureomycin. There was a direct correlation between amount of antibiotic and elevation of HD-1 LC₅₀s when 4-day-old larvae were used. LC₅₀ elevation in presence of antibiotic first appeared in 2-day-old larvae.     

Influence of COD/NH3–N ratio on organic removal and nitrification using a modified RBC

Synthetic wastewaters were prepared with different influent concentrations of ammonia nitrogen (NH₃–N) and COD and the treatment studies were conducted using a rotating biological contactor (RBC). If organic removal and nitrification can be simultaneously effected in one process, it will be an ideal solution to water pollution control. The RBC used in the present study was a four stage laboratory model and the discs were modified by attaching porous netlon sheets to enhance biofilm area. The COD loads (S ₀) used were about 1000 and 1500?mg/l whereas NH₃–N concentrations used were in the range of 20 to 185?mg/l. Hydraulic load (q) of 0.03?m³?.?m^-2?.?d^-1 and ammonia nitrogen loadings in the range of 0.66 to 5.5?g NH₃–N?.?m^-2?.?d^-1 were used. The RBC was operated at two different rotating speeds of 6 and 12?rpm. The results showed that the nitrification and percentage of COD removal were not affected up to the value of the COD/NH₃–N in the range from 47 to 23 at w=6?rpm and for an average influent COD of 1003?mg/l. Beyond that range only the nitrification rate decreased much whereas the percentage of COD removal was not affected. Similarly, at an influent COD load of 1557?mg/l, the nitrification and percentage COD removal were not affected for the value of the COD/NH₃–N in the range from 44 to 23 but beyond that range only the nitrification rate decreased while the percentage of COD removal was approximately constant and still high. A correlation plot between the NH₃–N removed and NH₃–N applied was presented at a rotating speed of 6?rpm and it was found that the nitrification rate of 3.93?g NH₃–N?.?m^-2?.?d^-1 was achieved at ammonia loading of 5.55?g NH₃–N?.?m^-2?.?d^-1. Also the results at w=12?rpm showed improvement of nitrification rate over those at 6?rpm.     

Ammonia toxicity as a criterion for the evaluation of larval quality in the prawn Macrobrachium rosenbergii

The feasibility of a short-term ammonia toxicity test as an evaluation criterion for larval quality was assessed in three trials. In each one, Macrobrachium rosenbergii larvae originating from the same spawn were nutritionally differentiated in two groups by feeding them either a nutrient-rich (Artemia nauplii enriched for 24 h with n-3 highly unsaturated fatty acids (HUFA) and ascorbic acid (AA)) or a nutrient-poor diet (Artemia nauplii starved for 24 h). Throughout their development, larvae from both treatments were exposed during 24 h to six concentrations of total ammonia (NH₄⁺+NH₃) and a control (no ammonia added). Based on mortality rates, the median lethal concentration for 50% of the population (LC₅₀) was estimated. As expected from earlier work, larvae fed the optimal diet presented higher n-3 HUFA and AA contents as well as higher growth and metamorphosis rates. From the moment the effect of diet quality was analytically detectable in the tissues of the larvae, the ammonia test was able to distinguish both groups of larvae. Differences in ammonia tolerance were observed as early as larval stage 4 and remained evident throughout larval development. The short-term ammonia toxicity test proved to be a valuable, sensitive and reproducible criterion for the establishment of larval quality.     

Exogenous sodium nitroprusside and glutathione alleviate copper toxicity by reducing copper uptake and oxidative damage in rice (Oryza sativa L.) seedlings

Nitric oxide (NO) and glutathione (GSH) regulate a variety of physiological processes and stress responses; however, their involvement in mitigating Cu toxicity in plants has not been extensively studied. This study investigated the interactive effect of exogenous sodium nitroprusside (SNP) and GSH on Cu homeostasis and Cu-induced oxidative damage in rice seedlings. Hydroponically grown 12-day-old seedlings were subjected to 100 μM CuSO₄ alone and in combination with 200 μM SNP (an NO donor) and 200 μM GSH. Cu exposure for 48 h resulted in toxicity symptoms such as stunted growth, chlorosis, and rolling in leaves. Cu toxicity was also manifested by a sharp increase in lipoxygenase (LOX) activity, lipid peroxidation (MDA), hydrogen peroxide (H₂O₂), proline (Pro) content, and rapid reductions in biomass, chlorophyll (Chl), and relative water content (RWC). Cu-caused oxidative stress was evident by overaccumulation of reactive oxygen species (ROS; superoxide (O₂ ^?–) and H₂O₂). Ascorbate (AsA) content decreased while GSH and phytochelatin (PC) content increased significantly in Cu-stressed seedlings. Exogenous SNP, GSH, or SNP?+?GSH decreased toxicity symptoms and diminished a Cu-induced increase in LOX activity, O₂ ^?–, H₂O₂, MDA, and Pro content. They also counteracted a Cu-induced increase in superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR), and glyoxalase I and glyoxalase II activities, which paralleled changes in ROS and MDA levels. These seedlings also showed a significant increase in catalase (CAT), glutathione peroxidase (GPX), dehydroascorbate reductase (DHAR), glutathione S-transferase (GST) activities, and AsA and PC content compared with the seedlings stressed with Cu alone. Cu analysis revealed that SNP and GSH restricted the accumulation of Cu in the roots and leaves of Cu-stressed seedlings. Our results suggest that Cu exposure provoked an oxidative burden while reduced Cu uptake and modulating the antioxidant defense and glyoxalase systems by adding SNP and GSH play an important role in alleviating Cu toxicity. Furthermore, the protective action of GSH and SNP?+?GSH was more efficient than SNP alone.     

Toxic Effect of Cadmium on Rice as Affected by Nitrogen Fertilizer Form

A nutrient solution experiment was conducted to determine the influence of N forms on growth, oxidative stress, and Cd and N uptake in rice plants. The treatments were consisted of two Cd levels (0 and 1 μmol) and three N forms (NH₄)₂SO₄, NH₄NO₃ and Ca(NO₃)₂. The results indicated that without Cd addition in the culture solution, the N forms had no significant effect on all measured parameters, including plant growth, photosynthetic traits, malondialdehyde (MDA) concentration, superoxide dismutase (SOD) activity, and Cd and N concentration, while Cd addition in the medium resulted in significant differences in measured parameters among the three forms of N fertilizers. The least inhibition of growth was noted in (NH₄)₂SO₄-fed plants, and the largest in Ca(NO₃)₂-fed plants, when plants were exposed to Cd stress. The highest photosynthetic rate and chlorophyll content was also recorded in (NH₄)₂SO₄-fed plants. Addition of Cd caused a remarkable increase in SOD activity and MDA content in plants, and the extent of increase varied with N form, with (NH₄)₂SO₄-fed plants being smallest. In comparison with the control plants, the N concentration in roots and shoots was not significantly affected in (NH₄)₂SO₄-fed plants, but significant decrease in root N concentration was found for the NH₄NO₃ and Ca(NO₃)₂-fed plants under Cd stress. Moreover, the significant differences were also noted among the three N forms in both root and shoot Cd concentrations, with (NH₄)₂SO₄-fed plants being the lowest. The results indicated that the toxic effect of Cd on rice varied with the form of N fertilizer.     

Caterpillar guts and ammonia volatilization: retention of nitrogen by gypsy moth larvae consuming oak foliage

The gypsy moth (Lymantria dispar L.), a major defoliator of hardwood forests in the eastern U.S., has a highly alkaline midgut pH. We hypothesized that the high pH would cause high rates of ammonia (NH₃) volatilization as larvae consumed foliage, leading to potentially large losses of N from the ecosystem to the atmosphere during gypsy moth outbreaks. We measured NH₃ emission during the consumption of oak foliage by larvae in the laboratory. Surprisingly, we found very low amounts of NH₃ release of about 0.1% of the N consumed in foliage. We speculate that digestive mechanisms may limit NH₃ production in the midgut, and that the acidic environment of the hindgut traps most of the small amount of NH₃ that is produced, effectively preventing a potentially very large N loss from both larvae and ecosystem. The estimated rate of NH₃ emission from a defoliated forest is small compared to other inputs and outputs of N from the ecosystem, but could potentially enhance the neutralization of atmospheric acidity during the defoliation period. Received: 12 May 1998 / Accepted: 28 July 1998     

Calcium and L-histidine effects on ascorbate-glutathione cycle components under nickel-induced oxidative stress in tomato plants

The effects of NiSO₄, calcium, and L-histidine (His) on the components of ascorbate-glutathione cycle, antioxidant enzymes and lipid peroxidation in a tomato cultivar Early Urbana Y was investigated. The activities of enzymes including catalase (CAT), guaiacol peroxidase (GPX), ascorbate peroxidase (APX), superoxide dismutase (SOD), glutathione reductase (GR), lipoxygenase (LOX), and phenylalanine ammonia lyase (PAL) were measured. In addition, the content of H₂O₂, ascorbate (ASC), dehydroascorbate (DHA), reduced glutathione (GSH), chlorophyll (Chl) a+b, carotenoids, proteins, malondialdehyde (MDA), membrane aldehydes, and electrolyte leakage (EL) were determined. Results suggest that the excess of Ni increased the content of H₂O₂, MDA, membrane aldehydes and proteins in roots as well as GPX, LOX, APX activities, and EL in leaves, whereas Ca and His ameliorated these effects. Moreover, decreasing leaf GSH and DHA content and GR activity were observed under the Ni stress, but these parameters were raised by Ca plus His treatment. However, no improvement in leaf protein, ASC, root GSH content, and activities of PAL and CAT were observed by using Ca or His under Ni stress.     

Propolis protects against high glucose-induced vascular endothelial dysfunction in isolated rat aorta

While propolis is known to have abundant bioactive constituents and a variety of biological activities, it is not clear whether propolis has beneficial effects on high glucose-mediated vascular endothelial impairment. The aim of the present study was to investigate the potential protective effect of propolis extract against the acute vascular endothelial dysfunction resulting from exposure to high glucose load and to elucidate its underlying mechanism. Rat aortic rings were incubated with normal glucose (11 mM), high glucose (44 mM), or mannitol (44 mM) for 3 h with or without propolis extract (400 μg/ml). Contraction to phenylephrine (Phe, 10^?9–10^?5 M) and relaxation to acetylcholine (ACh, 10^?9–10^?5 M) and sodium nitroprusside (SNP, 10^?9–10^?5 M) were measured before and after incubation. Changes in malondialdehyde (MDA), reduced glutathione (GSH), and superoxide dismutase (SOD) were also measured. Phe-induced contraction was impaired by high glucose as the E _max decreased from 138.87?±?11.43 to 103.65?±?11.5 %. In addition, ACh-induced relaxation was impaired as the E _max decreased from 99.80?±?7.25 to 39.20?±?6.5 %. SNP-induced relaxation was not affected. Furthermore, high glucose decreased the levels of both SOD (by 6 U/ml) and GSH (by 68 %) and increased levels of MDA (by 85 %). Propolis extract prevented high glucose-induced impairment of Phe and ACh responses and increased both SOD and GSH, leading to decreased MDA levels. In conclusion, propolis can protect against high glucose-induced vascular dysfunction by reducing oxidative stress.     

Nutrient limitation of Microcystis aeruginosa in northern California Klamath River reservoirs

Nutrient limitations were investigated in Copco and Iron Gate Reservoirs, on the Klamath River in California, where blooms of the toxin-producing cyanobacterium Microcystis aeruginosa were first reported in 2005. Nutrient enrichment experiments conducted in situ in June and August, 2007 and 2008, determined responses in phytoplankton biomass, Microcystis abundance and microcystin concentration to additions of phosphorus and different forms of nitrogen (NH₄⁺, NO₃, and urea). Microcystis abundance was determined using quantitative PCR targeting the phycocyanin intergenic spacer cpcBA.Total phytoplankton biomass increased with additions of N both before and during Microcystis blooms, with no primary effects from P, suggesting overall N limitation for phytoplankton growth during the summer season. NH₄⁺ generally produced the greatest response in phytoplankton growth, while Microcystis abundance increased in response to all forms of N. Microcystis doubling time in the in situ experiments was 1.24–1.39 days when N was not limiting growth. The results from this study suggest availability of N during the summer is a key growth-limiting factor for the initiation and maintenance of toxic Microcystis blooms in Copco and Iron Gate Reservoirs in the Klamath River.     

The effect of Myrtus communis L. ethanol extract on the small intestine and lungs in experimental thermal burn injury

Thermal trauma can damage organs away from the skin burn site and lead to multiple organ dysfunction. Following thermal injury, all tissues are exposed to ischemia, and as a result, resuscitation and reperfusion occur during the burning shock. Burn damage starts systemic inflammatory reactions that produce toxins and reactive oxygen radicals that lead to peroxidation. This study aimed to investigate, for the first time, the possible antioxidant effects of Myrtus communis ethanol extract on burn-induced oxidative distant organ injury orally. The thermal trauma was generated under ether anesthesia by exposing the dorsum of rats to 90 ^°C water bath for 10 s. 100 mg/kg/day Mrytus communis ethanol extract was applied orally for two days. Malondialdehyde (MDA) and glutathione (GSH) levels, glutatinone-S-transferase (GST), superoxidedismutase (SOD) and catalase (CAT) activities were determined to detect the possible antioxidant effects of myrtle on small intestine and lung tissues. Burn damage significantly increased MDA levels in lung and small intestine tissues, and significantly decreased GSH levels, CAT and GST activities in the small intestine and lung tissues compared to control group. Mrytus communis ethanol extract decreased MDA level and increased GSH level, SOD, CAT and GST activities significantly in either small intestine or lung tissues. Mrytus communis extract may be an ideal candidate to be used as an antioxidant adjunct to improve oxidative distant organ damage to limit the systemic inflammatory response and decreasing the recovery time after thermal injury.     

Hepatic and branchial glutathione S-transferases of two fish species: Substrate specificity and biotransformation of microcystin-LR

Liver and gills of roach (Rutilus rutilus) and silver carp (Hypophthalmichthys molitrix) were examined for glutathione S-transferases (GSTs) contents and their substrate specificity and capacity to biotransform microcystin-LR (MC-LR). GSTs and other glutathione (GSH) affine proteins were purified using a GSH-agarose matrix and separated by anionic chromatography (AEC). Substrate specificities were determined photometrical for 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), 4-nitrobenzyl chloride (pNBC) and ethacrynic acid (ETHA). Biotransformation rate of MC-LR was determined by high performance liquid chromatography (HPLC). Roach exhibited different hepatic and branchial GST activities for used substrates (DNB, pNBC and DCNB) compared to silver carp but not for ethacrynic acid. It suggests that, both fish species have similar amount of pi and/or alpha class, which were the dominant GST classes in liver and gills. Gills of both fish species contained a higher number of GST isoenzymes, but with lower activities and ability of MC-LR biotransformation than livers. GST isoenzymes from roach had higher activity to biotransform MC-LR (conversion rate ranging up to 268 ng MC-LR min^? 1 mL^? 1 hepatic enzyme) than that isolated from silver carp. Without any prior contact to MC-LR or another GST inducer, roach seems to be better equipped for microcystin biotransformation than silver carp.     

Ammonia-related metabolism of tobacco varieties differing in their susceptibility to Alternaria alternata

Some studies report that ammonia is an important factor of disease development in tobacco plants and various post-harvest fruits. Four tobacco (Nicotiana tabacum L.) varieties resistant or susceptible to Alternaria alternata (Fries) Keissler, a tobacco pathogenic fungus, were used to investigate whether there are differences in ammonia accumulation and the related metabolism of senescing leaves. The results showed that: (a) the leaves of susceptible varieties had significantly higher apoplastic [NH ₄ ⁺ ], pH, and ammonia emission potential (??-values) than resistant varieties during the period from 40 to 60 days of leaf age; (b) leaf tissue [NH ₄ ⁺ ] and total N concentrations in the tobacco varieties were not in line with their susceptibility or resistance to disease; (c) the increases in the apoplastic pH, ??-values, and leaf [NH ₄ ⁺ ] occurred in parallel with a significant decline in glutamine synthetase activity. Compared with the resistant varieties, apoplastic pH values and ?? values were increased more rapidly in the susceptible varieties due to a steeper decline in glutamine synthetase activity and a slower increase in glutamate dehydrogenase activity. In conclusion, NH₃ accumulation or NH3-dependent alkalinization rather than [NH ₄ ⁺ ] and total N appears to be mainly attributed to the enhanced susceptibility of tobacco plants to A. alternata.     

Stability of toxigenic Microcystis blooms

This is the first detailed study on the occurrence of cyanobacterial toxins in India, where we studied five eutrophic, temple ponds in the vicinity of Varanasi city, Uttar Pradesh, which continuously supported blooms of Microcystis sp. for several years. Bloom material from all five ponds was sampled bi-monthly from September 2003 to August 2004. Analysis of extracts by high-performance liquid chromatography (HPLC) indicated that microcystin-RR (MC-RR) was present all year round at high concentrations (311–1540 μg/g, dry weight), posing a significant health hazard especially since all five ponds are widely used for bathing, washing, cattle drinking supply, irrigation and recreation. In addition, there was unusually low temporal variation in concentration of MC-RR in each pond, <20% variation in four out of five ponds throughout the year.Characterization of microcystin composition of several bloom samples from this study by HPLC–PDA/MS confirmed that additional microcystins were present in many of the samples. The rarely reported, MC-AR was frequently detected in bloom samples from three of the ponds (Adityanagar, Durgakund and Sankuldhara), where it typically represented 20% of the microcystin pool. MC-WR was frequently found in samples from Adityanagar and Sankuldhara, representing 5–10% of the microcystin pool. MC-LR, along with the previously unreported MC-AHar, each represented approximately 5% of the microcystin pool when present. Bloom samples from each pond had a characteristic microcystin profile, when sampled from 2003 to 2006, suggesting persistent species/strain domination.The perennial and consistent nature of the toxic Microcystis blooms in these ponds is highly unusual, in contrast to the commonly encountered temporal and spatial variation of toxigenic and non-toxigenic species. Laboratory isolates from several ponds were shown to produce microcystins, showing similar microcystin composition to the original bloom material.     

Nitrogen dynamics at the sediment–water interface in shallow, sub-tropical Florida Bay: why denitrification efficiency may decrease with increased eutrophication

Nitrogen (N) dynamics at the sediment–water interface were examined in four regions of Florida Bay to provide mechanistic information on the fate and effects of increased N inputs to shallow, subtropical, coastal environments. Dissimilatory nitrate (NO₃ ^?) reduction to ammonium (DNRA) was hypothesized to be a significant mechanism retaining bioreactive N in this warm, saline coastal ecosystem. Nitrogen dynamics, phosphorus (P) fluxes, and sediment oxygen demand (SOD) were measured in north-central (Rankin Key; eutrophic), north-eastern (Duck Key; high N to P seston ratios), north-western (Murray Key; low N to P ratios), and central (Rabbit Key; typical central site) Florida Bay in August 2004, January 2005, and November 2006. Site water was passed over intact sediment cores, and changes in oxygen (O₂), phosphate (o-PO₄ ^3?), ammonium (NH₄ ⁺), NO₃ ^?, nitrite (NO₂ ^?), and N₂ concentrations were measured, without and with addition of excess ¹⁵NO₃ ^? or ¹⁵NH₄ ⁺ to inflow water. These incubations provided estimates of SOD, nutrient fluxes, N₂ production, and potential DNRA rates. Denitrification rates were lowest in summer, when SOD was highest. DNRA rates and NH₄ ⁺ fluxes were high in summer at the eutrophic Rankin site, when denitrification rates were low and almost no N₂ came from added ¹⁵NO₃ ^?. Highest ¹⁵NH₄ ⁺ accumulation, resulting from DNRA, occurred at Rabbit Key during a picocyanobacteria bloom in November. ¹⁵NH₄ ⁺ accumulation rates among the stations correlated with SOD in August and January, but not in November during the algal bloom. These mechanistic results help explain why bioreactive N supply rates are sometimes high in Florida Bay and why denitrification efficiency may decrease with increased NO₃ ^? inputs in sub-tropical coastal environments.     

Interaction of larval age and dietary formaldehyde on the susceptibility of tufted apple budmoth (Lepidoptera: Tortricidae)to Bacillus thuringiensis

Laboratory tests of Bacillus thuringiensis var. kurstaki (strain HD-1) against Platynota idaeusalis reared on a semisynthetic diet showed that the LC₅₀ values for neonates and 9-day-old larvae showed no significant difference in susceptibility; however, 14-day-old larvae were significantly more susceptible than neonates or 9-day-old larvae. There was no evidence of any interaction between 14-day-old larvae reared on the semisynthetic diets with and without formaldehyde and B. thuringiensis toxicity at any assessment reading. Likewise the toxicity of B. thuringiensis to 14-day-old larvae reared on apple leaves did not differ from the B. thuringiensis toxicity of 14-day-old larvae reared on semisynthetic diets with and without formaldehyde.     

Anti-Oxidative Defences Are Modulated Differentially in Three Freshwater Teleosts in Response to Ammonia-Induced Oxidative Stress

Oxidative stress and the antioxidant response induced by high environmental ammonia (HEA) were investigated in the liver and gills of three freshwater teleosts differing in their sensitivities to ammonia. The highly ammonia-sensitive salmonid Oncorhynchus mykiss (rainbow trout), the less ammonia sensitive cyprinid Cyprinus carpio (common carp) and the highly ammonia-resistant cyprinid Carassius auratus (goldfish) were exposed to 1 mM ammonia (as NH₄HCO₃) for 0 h (control), 3 h, 12 h, 24 h, 48 h, 84 h and 180 h. Results show that HEA exposure increased ammonia accumulation significantly in the liver of all the three fish species from 24 h–48 h onwards which was associated with an increment in oxidative stress, evidenced by elevation of xanthine oxidase activity and levels of hydrogen peroxide (H₂O₂) and malondialdehyde (MDA). Unlike in trout, H₂O₂ and MDA accumulation in carp and goldfish liver was restored to control levels (84 h–180 h); which was accompanied by a concomitant increase in superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase activity and reduced ascorbate content. Many of these defence parameters remained unaffected in trout liver, while components of the glutathione redox cycle (reduced glutathione, glutathione peroxidase and glutathione reductase) enhanced to a greater extent. The present findings suggest that trout rely mainly on glutathione dependent defensive mechanism while carp utilize SOD, CAT and ascorbate as anti-oxidative sentinels. Hepatic cells of goldfish appear to utilize each of these protective systems, and showed more effective anti-oxidative compensatory responses towards HEA than carp, while trout were least effective. The present work also indicates that HEA exposure resulted in a relatively mild oxidative stress in the gills of all three species. This probably explains the almost complete lack of anti-oxidative responses in branchial tissue. This research suggests that oxidative stress, as well as the antioxidant potential clearly differ between salmonid and cyprinid species.     

The Ammonia Transport, Retention and Futile Cycling Problem in Cyanobacteria

Ammonia is the preferred nitrogen source for many algae including the cyanobacterium Synechococcus elongatis (Synechococcus R-2; PCC 7942). Modelling ammonia uptake by cells is not straightforward because it exists in solution as NH₃ and NH ₄ ⁺ . NH₃ is readily diffusible not only via the lipid bilayer but also through aquaporins and other more specific porins. On the other hand, NH ₄ ⁺ requires cationic transporters to cross a membrane. Significant intracellular ammonia pools (≈1–10 mol?m^?3) are essential for the synthesis of amino acids from ammonia. The most common model envisaged for how cells take up ammonia and use it as a nitrogen source is the “pump–leak model” where uptake occurs through a simple diffusion of NH₃ or through an energy-driven NH ₄ ⁺ pump balancing a leak of NH₃ out of the cell. The flaw in such models is that cells maintain intracellular pools of ammonia much higher than predicted by such models. With caution, [¹⁴C]-methylamine can be used as an analogue tracer for ammonia and has been used to test various models of ammonia transport and metabolism. In this study, simple “proton trapping” accumulation by the diffusion of uncharged CH₃NH₂ has been compared to systems where CH₃NH ₃ ⁺ is taken up through channels, driven by the membrane potential (ΔU _i,o) or the electrochemical potential for Na⁺ (ΔμNa _i,o ⁺ ). No model can be reconciled with experimental data unless the permeability of CH₃NH₂ across the cell membrane is asymmetric: permeability into the cell is very high through gated porins, whereas permeability out of the cell is very low (≈40 nm?s^?1) and independent of the extracellular pH. The best model is a Na _in ⁺ /CH₃NH ₃ ⁺ _in co-porter driven by ΔμNa _i,o ⁺ balancing synthesis of methylglutamine and a slow leak governed by Ficks law, and so there is significant futile cycling of methylamine across the cell membrane to maintain intracellular methylamine pools high enough for fixation by glutamine synthetase. The modified pump–leak model with asymmetric permeability of the uncharged form is a viable model for understanding ammonia uptake and retention in plants, free-living microbes and organisms in symbiotic relationships.     

Absence of negative allelopathic effects of cylindrospermopsin and microcystin-LR on selected marine and freshwater phytoplankton species

Cyanobacterial toxins have been regarded by some researchers as allelopathic substances that could modulate the growth of competitors. Nevertheless, often the concentrations of toxins used are too high to be considered ecologically relevant. In this work we tested the hypothesis that microcystin-LR (MC-LR) and cylindrospermopsin (CYN) at ecologically relevant concentrations have no allelopathic effects on some species of phytoplankton. Extracts containing the toxins as well as pure MC-LR and CYN toxins were used to assess their effects on the growth rates of Nannochloropsis sp., Chlamydomonas reinhardtii, and Chlorella vulgaris. Cyanobacterial crude extracts induced more pronounced effects on growth rates than pure toxins. Microcystis aeruginosa and Aphanizomenon ovalisporum crude extracts containing MC-LR and CYN at 0.025–2.5 mg l^?1 stimulated growth rates of microalgae, whereas A. ovalisporum crude extracts containing 2.5 mg l^?1 of CYN strongly inhibited growth rates of microalgae after 4 and 7 days of exposure. MC-LR and CYN at environmentally occurring concentrations were unable to affect negatively the growth of microalgae, and therefore these molecules may play roles other than allelopathy in natural ecosystems.