Bioaccumulation and toxic effects of copper on growth and oxygen consumption by the postlarvae of Penaeus indicus

The aim of this study was to assess the toxic impact of copper on postlarvae (PL) of the penaeid shrimp Penaeus indicus. Tolerance, growth, oxygen consumption and metal accumulation were investigated in these PL on exposure to copper. Tolerance studies were conducted for 96 h to assess the tolerance limits of P. indicus PL exposed to different concentrations of copper using static renewal bioassay tests. Using the Probit method, the regression equation was calculated as Y=0.4899+2.3562 X, with a correlation coefficient of 0.9707. The 96 h LC₅₀ was 0.8204 ppm. The effect of sublethal (one-fifth of 96 h LC₅₀) copper on PL for short- and long-term exposures revealed a significant (p<0.05) decrease in the rate of oxygen consumption, metabolic rate, mean length, wet and dry weight of the exposed PL over their respective controls which can be attributed to a gradual and time-dependent accumulation of the metal, as noticed in the exposed PL through accumulation studies. Overall, the data suggest that on chronic exposure even sublethal concentrations of copper can reduce the metabolic rate and growth in P. indicus PL. This is perhaps the first attempt to use the wild P. indicus PL as a bioindicator of copper toxicity.     

Temperature effects on the toxicity of four trace metals to adult spotted Babylonia snails (Babylonia areolata)

A study was conducted to determine the median lethal toxicity of four heavy metals on the marine gastropod Babylonia areolata. Median lethal toxicity tests were conducted to observe the sensitivity of this gastropod to metals and how variations in temperature might affect toxicity of test elements. Four heavy metals were used in the study. It was observed that the 96-hr LC₅₀ (in mg/L) for the different metals was found to be nickel (Ni) 33.53 (35.22–28.43), copper (Cu) 44.59 (46.43–41.53), cadmium (Cd) 21.53 (23.43–18.37), and zinc (Zn) 27.34 (28.81–24.24) at room temperature 24 °C. With temperature as a variable, median lethal concentration (LC₅₀) values were observed to increase from 22.41 mg/L at 10 °C to 27.34 mg/L at 28 °C and reduce to 18.43 mg/L at 30 °C and a further rise in toxicity was observed at 35 °C where LC₅₀ value was 12.7 mg/L as seen in the case of Zn. It was also observed that at 40 °C thermal and chemical toxicity overlapped as 100% mortality was observed in controls. This trend was noted in all metals for Babylonia areolata indicating that temperature played an important role in determining LC₅₀ values of toxicants.     

Effects of salinity and possible interactions with temperature and pH on growth and photosynthesis of <Emphasis Type="Italic">Halophila johnsonii</Emphasis> Eiseman

The effects of salinity, temperature, and pH variations on growth, survival, and photosynthetic rates of the seagrass Halophila johnsonii Eiseman were examined. Growth and survival responses to salinity were characterized by aquarium experiments in which plants were exposed to seven different salinity treatments (0, 10, 20, 30, 40, 50, and 60 psu) during 15 days. Photosynthetic behavior was assessed for short-term salinity exposures (1 or 20 h) by incubation experiments in biological oxygen demand (BOD) bottles and by measuring photosynthesis versus irradiance (PI) responses in an oxygen electrode chamber. In the bottle experiments the possible effects of interactions between salinity and temperature (15, 25, and 35°C) or pH (5, 6, 7, and 8.2) were also examined. Growth and survival of H. johnsonii were significantly affected by salinity, with maximum rates obtained at 30 psu. Salinity also altered the parameters of the PI curves. Light-saturated photosynthesis (P _max) and the photosynthetic efficiency at subsaturating light (α) increased significantly up to an optimum of 40 psu, decreasing again at the highest salinities. Dark respiration rates and compensating irradiance (I _c) showed minimum values at 40 and 50 psu, while light-saturation point (I _k) was maximum at 30–50 psu. An interaction between salinity and temperature was not found although an increase of temperature alone produced an increase in α, P _max, respiration rates, and I _k. An interaction between salinity and pH was only found in the P _max response: P _max increased with pH=5 at 30 psu. In addition, reducing the pH increased α significantly. In the BOD bottles experiment a significant reduction in the dark respiration with decreasing pH was observed, but the opposite trend was observed in the photosynthetic rate. These results suggest that the endemic seagrass H. johnsonii could be negatively affected by hypo- or hypersalinity conditions, although salinity changes did not seem to alter the tolerance of this species to other environmental factors, such as temperature or pH.     

Influence of some abiotic environmental factors on acute toxicity of inorganic lead to Cyprinus carpio var communis (Linn.) and Catla catla (Ham.) in simulated toxic aquatic environment

With the increase of water hardness from 60 to 720?mg/L CaCO₃, total alkalinity from 32 to 376?mg/L CaCO_3, pH from 7.6 to 7.9 and chloride from 28 to 350?mg/L, 96?h LC₅₀ on the basis of total lead increased from 8.2 to 1291?mg/L for Cyprinus carpio and 5.3 to 865?mg/L for Catla catla, when soil sediments were included these values were further raised to 1356 and 874?mg/L, respectively. The dissolved lead LC₅₀ values in all the treatments of soil and water was consistent with fixed amount of dissolved lead (1.04–1.78?mg/L) being needed for median lethal toxicity. Total lead toxicity also decreased with increase in pH from 6.3 to 11.3. 96?h LC₅₀ values increased for common carp 15 to 631?mg/L and for catla, 8 to 355?mg/L. But dissolved lead toxicity was found to increase with the increase of pH from 6.3 to 11.3 for both common carp (LC₅₀, 3.53 to 0.24?mg/L) and catla (LC₅₀, 2.21 to 0.09?mg/L). Removal of dissolved Pb with increasing carbonate content, particulate matter and pH due to adsorption, precipitation or coprecipitation reaction, reduced the dissolved lead concentration and thus the total lead toxicity. Increase in toxicity of lead with increase of exposure time was the biological response of longer contact time and decrease in dissolved lead toxicity with decrease in pH was due to increase H⁺ ion competition.     

Monitoring toxicity of an azo dye methyl red and a heavy metal Cu,using plant and animal bioassays

Toxicities of an azo dye methyl red and a heavy metal copper (Cu) were quantified, using growth and mortality as end points, in four plant species and three animal species by subjecting them to short-term (4 days for animals, 10 days for plants) static bioassays. Lemna aequinoctialis Welwitch (EC₅₀: 7–16 ppm) was found to be the most sensitive species for methyl red, Ceratophyllum demersum L. (EC₅₀: 25 ppm) and Lactuca sativa L. (EC₅₀: 56 ppm) were intermediate, while Oryza sativa L. shows reduction in seedling vigor (9–27%) of <50%, being the least sensitive amongst the tested plant species. Methyl red toxicity is almost 3–5-fold higher in growing medium (pH = 5.8–6.0), even at high nutrient levels, while Cu toxicity is higher in nutrient-poor alkaline medium at alkaline pH (8.3–8.7; EC₅₀: Ceratophyllum = 104–200 ppb; Lemna = 100–170 ppb) compared to nutrient-rich acidic medium (pH = 5.4–5.7; EC₅₀: Ceratophyllum = 2600–3175 ppb; Lemna = 4350–4715 ppb). Rice tolerance (EC₅₀: 6500 ppb) was found to be higher than hydrophytes while lettuce was most tolerant to Cu. Fish sensitivity toward the test chemicals was almost parallel to Ceratophyllum and Lemna [Gambusia affinis Baird and Gerard (LC₅₀: 250 ppb for Cu) and Poecilia reticulata Peters (LC₅₀: 24 ppm for methyl red)]. Similar to the plants, dye toxicity increased markedly (LC₅₀: 7 ppm) in the acidic medium (pH = 6.0). Amongst the tested organisms, Daphnia was found to be most sensitive to methyl red (EC₅₀: 6 ppm) while its sensitivity to Cu (EC₅₀: 230 ppb) was similar to hydrophytes and fish. Initially, the combination of dye and Cu (at their sublethal concentrations) had additive effects in duckweed, while dye concentration ruled afterward. These results indicate that hydrophytes and animals are equally sensitive toward the test chemicals. Dye toxicity in hydrophytes and fish was pH dependent, while in the case of Cu, it is related to the nutrient status of the growth medium of plants.     

Acute toxicity evaluation of triazophos,deltamethrin and their combination on earthworm,Eudrilus eugeniae and its impact on AChE activity

The acute toxicity of three formula grade pesticides namely, triazophos (an organophosphate, OP), deltamethrin (a pyrethroid) and combined pesticide (triazophos?+?deltamethrin) was determined in earthworm, Eudrilus eugeniae. They were exposed to different concentrations of these pesticides for 48 h by paper contact toxicity method. The LC₅₀ values for triazophos, deltamethrin and of combination were determined as 0.076, 0.031 and 0.065?μg/cm², respectively. To study the sublethal effect of these pesticides, E. eugeniae were exposed to 5% and 10% of LC₅₀ of pesticides for 48 h. Morphological alterations such as coiling, clitellar swelling, mucus release and bleeding followed by body segmentation were observed in exposed earthworm. Acetylcholinesterase (AChE) activity assayed in different regions of body segment exhibited a significant (p?<?0.05) decrease in its activity particularly in the pre-clitellar region as compared to other regions. The altered behavioural responses in pesticides exposed earthworms would have been due to decline in AChE activity of the nervous system.     

Acute toxicity of sugarcane vinasse to aquatic organisms before and after pH adjustment

The purpose of this study was to investigate the toxicity of vinasse to cladocerans and fish before and after pH adjustment using an acute toxicity test. Linear and quadratic regression models were adjusted to demonstrate the concentration–response relationship between vinasse and the endpoints evaluated. The median lethal concentrations (LC_50–48h) of vinasse before pH adjustment for Ceriodaphnia dubia and Daphnia magna were 0.67% and 0.80% respectively, and the median lethal concentrations (LC_50–96h) for Danio rerio was 2.62%. After pH adjustment, the values increased for all organisms, demonstrating a decrease in toxicity. This study reported marked toxicity for vinasse to aquatic organisms with toxicity reduction after pH adjustment.     

Acute and chronic toxic effect of lead (Pb) and zinc (Zn) on biomarker response in post larvae of Penaeus monodon (Fabricus, 1798)

Metal pollution produces damage to marine organisms at the cellular level possibly leading to ecological imbalance. The present investigation focused on the acute and chronic toxicity of lead (Pb) and zinc (Zn) by examining the effects of biomarker enzymes in post-larvae of Penaeus monodon (Tiger prawn). Antioxidant biomarker responses such as lipid peroxidation (LPO) and catalase (CAT) activity for Pb and Zn were determined following chronic exposure. Acute Lethal Concentration ₅₀ (LC₅₀) values observed in the study at 96?h for Pb and Zn at 5.77?±?0.32?mg?L^?1 and 3.02?±?0.82?mg?L^?1, respectively. The estimated No Observed Effect Concentration and Lowest Observed Effect Concentration values for Pb were 0.014 and 0.029?mg?L^?1 and that recorded for Zn was 0.011 and 0.022?mg?L^?1, respectively. Among the two metals studied, toxicity of Zn was found to be greater to P. monodon than Pb. The activities of antioxidant defense enzymes and total protein content differed significantly from control following exposure to both metals. Overall, the biomarker studies demonstrated that alterations in antioxidant enzymes and induction of LPO reflect the consequences of heavy metal exposure in P. monodon.     

The Influence of Temperature and Salinity Upon the Acute Toxicity of Heavy Metals to the Banana Prawn (Penaeus merguiensis de Man)

Bioassays were conducted to determine the effects of temperature and salinity on the acute toxicity of mercury, copper, cadmium, zinc, nickel and lead to juvenile banana prawns (Penaeus merguiensis de Man). Tests were conducted at all combinations of 35, 30 and 20°C with 36 and 20° salinity over 96 h. The general rank order of metal toxicity was Hg > (Cu, Cd, Zn) > Ni > Pb. The toxicity of all metals increased with increased temperature. This was most noticeable in the high salinity treatments, particularly for copper and zinc. Salinity appeared to influence the toxicity of all metals tested although significant differences were only found for copper and lead at 20°C. At this temperature prawns were markedly more susceptible to both metals in low salinity sea water. The data are compared with lethal concentrations found for other crustaceans and are discussed in relation to existing water quality criteria.     

The influence of solid matrices on arsenic toxicity to Corophium orientale

In marine ecosystems, benthic organisms are really important because they are the first step in the transfer of contaminants from environment to biota. To this end, this study focused on biological assays with the amphipod Corophium orientale exposed to two different molecules of arsenic: arsenate (As^V), the most abundant form in sediments, and dimethyl-arsinate (DMA), expected to be moderately toxic as an intermediate in the process of detoxification. The toxicity of arsenic compounds was measured after exposure to three different matrices: water, spiked natural sediment and inert spiked quartz sand. LC₅₀ values were calculated for each exposure, and the results confirmed the highest toxicity of As^V, in addition to underlining the importance of matrix of exposure. Water exposure was the matrix which presented the highest toxicity for inorganic arsenic (As^V LC₅₀=3.51 mg L^?1 vs DMA LC₅₀=54.65 mg L^?1), spiked natural sediment demonstrated its capability to chelate arsenate toxicity (As^V LC₅₀=34.27 mg kg^?1 vs. DMA LC₅₀=52.19 mg kg^?1) and spiked quartz sand presented intermediate values for As^V (LC₅₀=25.26 mg kg^?1), whereas for DMA a lower toxicity was registered (LC₅₀=872.35 mg kg^?1). This study can provide some useful data linked with chemical speciation of arsenic and exposure matrix, for improving the correct management of contaminated sediment.     

Metabolic response under different salinity and temperature conditions for glass eel Anguilla japonica

Diadromous fish often enter freshwater directly from seawater via fish ladders or channels built in estuarine dams. The oxygen consumption rates (OCR) of glass eel, Anguilla japonica, were determined using an automatic intermittent flow respirometer under various salinity and temperature regimes to physiologically explain this direct movement. The endogenous rhythm of the OCR in wild glass eels, freshly collected from estuaries, was nearly synchronous with the tidal pattern at the estuarine collection site. When the salinity was changed from 20 psu (12°C) at a constant temperature to that of freshwater, the OCR of the glass eels decreased by 21.6±7.0% (mean ± SD) (P<0.05), showing a dampened rhythm for about 48 h. After this period of impediment, the glass eels resumed normal metabolic activity. Direct migration from seawater to freshwater under constant temperature would result in a severe physiological stress for these glass eels for about two days. When the glass eels were exposed to a cyclic change in water temperature of 2°C 26 h⁻¹, as they encounter in estuaries, and then were introduced to freshwater abruptly, the OCR rhythm corresponded to the cyclic changes in water temperature after exposure to freshwater. Under these conditions, the mean OCR of the glass eels had a small difference before and after exposure to freshwater. These data explained how glass eels can directly move from sea water into the freshwater without any apparent metabolic stress in the estuaries showing cyclic change in water temperature (Δt=2°C).     

Growth and development of nauplii and copepodites of the estuarine copepod Acartia tonsa from southern Europe (Ria de Aveiro, Portugal) under saturating food conditions

A temperature-dependent growth model is presented for nauplii and copepodites of the estuarine calanoid copepod Acartia tonsa from southern Europe (Portugal). Development was followed from egg to adult in the laboratory at four temperatures (10, 15, 18 and 22°C) and under saturating food conditions (>1,000 μg C l⁻¹). Development times versus incubation temperature were fitted to a Belehradek’s function, showing that development times decreased with increasing incubation temperature: at 10°C, A. tonsa need 40.3 days to reach adult stage, decreasing to 8.9 days when reared at 22°C. ANCOVA (homogeneity of slopes) showed that temperature (P<0.001) and growth phase (P<0.01) had a significant effect on the growth rate. Over the range of temperatures tested in this study, highest weight-specific growth rates were found during naupliar development (NI–NVI) and varied from 0.185 day⁻¹ (10°C) to 0.880 day⁻¹ (22°C) with a Q ₁₀ equal to 3.66. During copepodite growth (CI–CV), the weight-specific growth rates ranged from 0.125 day⁻¹ (10°C) to 0.488 day⁻¹ (22°C) with a Q ₁₀ equal to 3.12. The weight-specific growth rates (g) followed temperature (T) by a linear relationship and described as ln g=−2.962+0.130 T (r ²=0.99, P<0.001) for naupliar stages and ln g=−3.134+0.114T (r ²=0.97, P<0.001) for copepodite stages. By comparing in situ growth rates (juvenile growth and fecundity) for A. tonsa taken from the literature with the temperature-dependent growth model defined here we suggest that the adult females of A. tonsa are more frequently food limited than juveniles.     

Studies of thermodynamics of phosphomidon on fly ash

Adsorption thermodynamic studies of phosphomidon on fly ash at 25° and 50°C have been analysed as adsorption isotherms, Freundlich equations, K_d values and various other thermodynamic parameters. These data were in close agreement with Freundlich isotherms and yield ‘S’ type isotherms at both the temperatures. Thermodynamic constants (K_o) and standard free energy (ΔG°), enthalpy (ΔH°) and entropy changes (ΔS°) have been calculated for predicting the nature of adsorption.     

Effects of temperature and salinity acclimation of adults on larval survival,physiology, and early development of Lytechinus variegatus (Echinodermata: Echinoidea)

Larval survival and developmental rates of Lytechinus variegatus (Lamarck) were determined as a function of temperature and salinity in two experiments by: (1) directly transferring fertilized eggs to 35, 30, 27.5, 25, 20, 15, and 10S seawater at 18 and 23°C, and (2) acclimation of adult sea urchins to the conditions described above for 1 to 4 wk prior to spawning. Developmental rates and percent survival of larvae prior to metamorphosis decreased at salinities below 35 (Q₁₀ values for metamorphosis=0.380 to 0.384). Temperature and salinity significantly (P<0.05) affected metabolic rates of L. variegatus plutei. These results show that L. variegatus larvae are stenohaline when compared to larvae of other echinoderm species. LC₅₀ values (S), developmental rates, and survival to metamorphosis indicate that acclimation of adult sea urchins to lower salinity prior to spawing and fertilization does not enhance development or survival of embryos exposed to low salinity.     

Effects of ammonia on survival and osmoregulation of various development stages of the shrimp Penaeus japonicus

In Penaeus japonicus, the tolerance to ammonia increased with the development from nauplius to late juvenile. The 48-h LC₅₀ of ammonia in nauplii (III–V), 96-h LC₅₀ in zoeae (I–III), mysis (I–III), post-larvae (PL1) and late juveniles (10.4±1.1 g) were respectively 5.0, 6.1 to 8.1, 9.4 to 10.9, 15.5 and 52.7 mg Nl^-1 (0.5, 0.6 to 0.7, 0.9, 1.3 and 3.1 mg NH₃–Nl^-1). In a chronic experiment (20 d), the LC₅₀ in post-larvae (PL1) was 19.1 (1.4) at 96 h and 16.2 mg Nl^-1 (1.3 mg NH₃–Nl^-1) at 480 h. Osmoregulatory capacity (OC) was calculated as the osmotic gradient between the hemolymph and the external medium at given salinities. The effects of ammonia on OC, Na⁺ and Cl^- regulation and gill Na⁺–K⁺ ATPase activity in late juveniles were examined in fullstrength seawater, SW (1050 mosm kg^-1, 36 S) and in dilute SW (450 mosm kg^-1, 15%.), after 48 or 96 h exposure to various concentrations of ammonia. Ambient ammonia disrupted both hypo- and hyper-osmoregulation; decreased OC resulted from impaired Na⁺ and Cl^- regulation. Gill Na⁺–K⁺ ATPase activity increased in SW and was not affected in dilute SW. The decrease of OC was ammonia-dose-dependent. The threshold ammonia concentrations affecting hypo-OC and hyper-OC were, respectively, 16 (1.3) and 32 mg Nl^-1 (2.3 NH₃–Nl^-1) for a 48 h exposure; these concentrations were lower than the 48-h LC₅₀ value, 65.3 mg Nl^-1 (3.5 NH₃–Nl^-1). The time course of exposure to sublethal ammonia (48 mg Nl^-1) demonstrated that the effect on osmoregulation was time-dependent. This effect was also temporary, and the exposed shrimps recovered control OC values after removal of excessive ambient ammonia. The possibility of using OC as an indicator of physiological condition in osmoregulating crustaceans and the acting mode of ammonia on osmotic and ionic regulation are discussed.     

Individual and combined toxicity of some petroleum aromatics to the marine amphipod Elasmopus pectenicrus

Toxicity of 4 components of petroleum oils to the marine amphipod Elasmopus pectenicrus (Bate) has been assessed. Two ephemeral aromatic hydrocarbons, naphthalene (A) and 1, 2, 4-trimethylbenzene (B) were more toxic than two persistent aromatics, o-cresol (C) and o-toluidine (D). The acute toxicity concentrations obtained for individual aromatic compounds were always greater than the actual concentrations found in the water-soluble fractions (WSF) of fuel oils. Results from mixtures of 2 or more components indicated that the LC₅₀ levels were primarily determined by the more toxic substances, A and B. Naphthalene and 1, 2, 4-trimethylbenzene became more toxic to the E. pectenicrus when present in a mixture of more than 2 components, and the toxicity increased with increasing numbers of components present. Synergistic effects, therefore, possibly occur in the whole WSF. No antagonistic effects were observed among the 4 petroleum aromatics.University of Texas, Marine Science Institute Contribution No. 290     

Survival and growth of bivalve larvae under heavy-metal stress

In a study of the toxicity of mercury, silver, copper, nickel, and zinc to larvae of the American oyster Crassostrea virginica and hard clam Mercenaria mercenaria, the concentrations at which 5% (LC₅), 50% (LC₅₀), and 95% (LC₉₅) of the larvae died were determined, as well as growth at the LC₅ and LC₅₀ values. The order of toxicity for oyster larvae was Hg>Ag>Cu>Ni, and for clam larvae Hg>Cu>Ag>Zn>Ni. Growth of larvae of both species, with the exception of clam larvae in nickel-treated water, was not reduced at the LC₅ values, but was markedly reduced at the LC₅₀ values.     

Differential toxic response of Chironomus travalensis kieffer ( Chironomidae : Diptera ) to some industrial effluents

Acute toxicity testing of some industrial effluents (rubber (Rr), brewery (Br), and bottling (Bt)) on Chironomus travalensis (Diptera larvae) was determined at 96-h using a static bioassay technique. Effluent quality characterization revealed acidic pH values of 4.5 for Rr, 5.64 for Br, and 4.32 for Bt effluents. Tests were carried out in replicates between 10 and 100% effluent concentrations. Mortality rate rose progressively with increasing concentrations. Bt effluent was the most toxic with 96-h LC₅₀ of 61.5%, closely followed by Rr effluent with 96-h LC₅₀ of 87% and the least toxic was Br effluent with 96-h LC₅₀ of 90%.     

Removal of cadmium(II), lead(II), and chromium(VI) ions from aqueous solution using clay

Removal of cadmium(II), lead(II), and chromium(VI) from aqueous solution using clay, a naturally occurring low-cost adsorbent, under various conditions, such as contact time, initial concentration, temperature, and pH has been investigated. The sorption of these metals follows both Langmuir and Freundlich adsorption isotherms. The magnitude of Langmuir and Freundlich constants at 30°C for cadmium, lead, and chromium indicate good adsorption capacity. The kinetic rate constants (K _ad) indicate that the adsorption follows first order. The thermodynamic parameters: free energy change (ΔG ^o), enthalpy change (ΔH ^o), and entropy change (ΔS ^o) show that adsorption is an endothermic process and that adsorption is favored at high temperature. The results reveal that clay is a good adsorbent for the removal of these metals from wastewater.     

Thermal dependency of burrowing in three species within the bivalve genus <Emphasis Type="Italic">Laternula</Emphasis>: a latitudinal comparison

The upper thermal limits for burrowing and survival were compared with micro-habitat temperature for anomalodesmatan clams: Laternula elliptica (Antarctica, 67°S); Laternula recta, (temperate Australia, 38°S) and Laternula truncata (tropical Singapore, 1°N). Lethal limits (LT₅₀) were higher than burrowing limits (BT₅₀) in L. elliptica (7.5–9.0 and 2.2°C) and L. recta (winter, 32.8–36.8 and 31.1–32.8°C) but the same range for L. truncata (33.0–35.0 and 33.4–34.9°C). L. elliptica and L. truncata had a BT₅₀ 0.4 and 2.4–3.9°C, respectively, above their maximum experienced temperature. L. recta, which experience solar heating during midday low tides, had a BT₅₀ 0.7–2.4°C below and a range for LT₅₀ that spanned their predicted environmental maximum (33.5°C). L. recta showed no seasonal difference in LT₅₀ or BT₅₀. Our single genus comparisons contrast with macrophysiological studies showing that temperate species cope better with elevated temperatures. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.