A sensitive spectrophotometric method for the determination of arsenic in environmental samples

We developed a cost-effective and sensitive spectrophotometric method for the determination of arsenic at trace level using a new reagent, leuco malachite green. Here we show that, arsenic reacts with potassium iodate in acidic conditions to liberate iodine, and the liberated iodine selectively oxidizes leuco malachite green to malachite green dye. We studied the Beer’s law at 617 nm, which showed linearity over the concentration range 0.09–0.9 μg ml⁻¹ of arsenic. We show that the molar absorptivity, Sandell’s sensitivity and detection limit of the method are 6.1 × 10⁴ l mol⁻¹ cm⁻¹, 0.0012 μg cm⁻² and 0.025 μg ml⁻¹, respectively. We applied the developed method for the determination of arsenic in environmental samples.     

A rapid kinetic technique for quantifying polycarbonate species in coalspoils

A rapid, automated procedure has been used for differentiation and quantitative determination of siderite, calcite and dolomite, based on the different rates of dissolution of the three carbonate species when reacted with excess 5 mol/litre HCl. The method employs manometric measurements of acid-liberated CO₂ with a membrane differential-pressure transducer coupled to a stripchart recorder. Tested on the three carbonate species, singly and in pairs, the technique is discriminatory, sensitive and relatively rapid.Contributions from the Department of Agronomy, University of Kentucky Agricultural Experiment Station in Lexington, and Institute for Mining and Minerals Research of the University of Kentucky. The investigation reported in this paper (No. 83-3-206) is in connection with a project of the Kentucky Agricultural Experiment Station and is published with the approval of the Director.     

A nanosilver-based spectrophotometric method for sensitive determination of methyl violet in river water

A method is reported for the determination of methyl violet in the range of 10–120 nmol L^?1. The method is based on the catalytic effect of silver nanoparticles (AgNPs) on the oxidation reaction of methyl violet by potassium bromate in acid medium. The reaction is followed spectrophotometrically by measuring the change in absorbance () at 620 nm using a fixed time method. The reaction variables were optimized in order to achieve highest sensitivity. The 3б criterion detection limit was 5 nmol L^?1, and the relative standard deviation for ten replicate determinations at a concentration of methyl violet of 15 nmol L^?1 was 0.97% (n = 10). The method was successfully applied to the determination of methyl violet in river water samples.     

A radiotracer technique for the determination of male mating success in natural populations

Summary Males of a natural population of Antechinus stuartii (Marsupialia) were injected at the beginning of their short, synchronous breeding period with one of twelve gamma-emitting nuclides which are individually recognizable by their unique spectial properties. This label passed to the females during ejaculation and was identified when the females were captured and counted in a whole-body counter. This technique established mating success of males and provided an estimate of reproductive success.     

A new application of the diacetyl monoxime method to the automated determination of dissolved urea in seawater

A new application of the diacetyl monoxime method for the determination of dissolved urea in seawater is presented, based on the Alpkem autoanalyzer (OI, Analytical). This system has several advantages over automated methods previously described in the literature. The urea cartridge can be easily built using nut/ferrule connections, and the external thermostatic bath is not required. The system is set up to operate at low urea concentrations (0–3 µM urea), but it is linear up to 100 µM urea. The sample volume needed for analysis is 0.4 ml, and the rate of analysis is 25 samples h^–1. The precision of analysis (CV) of seawater samples is 2.1% at 0.50 µM urea, and the detection limit is 0.02 µM urea. The interference on the colorimetric measurement due to formation of Brij-35 aggregate in strongly acidic media was eliminated by substituting this wetting agent with hexadecyl–trimethyl–ammonium bromide; thus, this method is suitable for other autoanalyzer technologies that use small-diameter transmission tubes and small-sized spectrophotometric flowcells.Communicated by R. Cattaneo-Vietti, Genova     

Rapid and sensitive headspace gas chromatographic method for cyanide determination in whole blood

We reported a simple and rapid headspace (HS) gas chromatographic (GC) assay for the determination of cyanide in human whole blood obtained from both cadaveric and intoxicated subjects. The poison was extracted from whole blood samples by HS technique after 10 min incubation at 80°C with a HCl (1N) acidic solution supplemented with 30 g L^?1 of sodium chloride and 10 mg L^?1 of isopropanol as internal standard. A SGE WAX capillary column (30 m × 0.32 mm ID × 0.5 µm df) was used. The Flame Ionization Detector (FID) was set at 250°C. The detection and the quantification limits were 0.02 and 0.2 mg L^?1, respectively. In the within-day study, the coefficients of variation for three different whole blood cyanide concentrations varied between 3.2 and 6.5%, whereas those in the day-to-day study varied between 4.8 and 6.3%. The assay was performed in less than 20 min per sample, which appears to be suitable for emergency toxicological tasks involved in diagnosis of cyanide poisoning.     

A recirculating seawater aquarium system for inland laboratories

We have designed and installed a 7,500 l recirculating aquarium system for the purpose of maintaining marine gastropods in an inland laboratory. It is completely nonmetallic and can be adapted for any number of isolated living compartments. Vonvenient and efficient filtration is achieved by the use of disposable cartridge filters. The water temperature is regulated by a direct expansion cooling coil and a hot water heating coil without the intervention of secondary heat transfer fluids. Most of the water is stored in an underground reservoir which contributes to thermal stability, and the water temperature varies <±0.1°C per week. The pH and specific gravity remain at 8.0±0.1 and 1.023+0.002-0.001, respectively, for as long as 4 months. We have no evidence that any animal (Aplysia, Hermissenda, Tritonia) has died due to poor water quality. The system is constructed from readily available components, is moderate in cost, and has performed reliably for over 2 years.     

An empirical test of biases in the rapid visual technique for species-time censuses of reef fish assemblages

For an assemblage of fishes inhabiting a warm temperate, rock reef near San Diego, California (USA), a series of censuses were made based on frequency of occurrence and standard numerical counts. These data were then compared with a parallel study based on the rapid visual technique (RVT) of Jones and Thompson (1978). Because the RVT ranks abundances according to frequency of encounter and disregards variations in the spatial distributions of different species, the method overemphasizes the importance of widespread albeit rare fishes but under-emphasizes patchy although abundant species. The discussion considers the relative merits of the RVT method for characterizing assemblages of fishes on temperate rock reefs and on tropical coral reefs.     

Quantification of trypsin with a radioimmunoassay in herring larvae (Clupea harengus) compared with a highly sensitive fluorescence technique to determine tryptic enzyme activity

Enzymatic activity and quantity of the protease trypsin were measured in individual herring larvae (Clupea harengus L.). The enzymatic activity assay was done using a fluorescence technique, and a radioimmunoassay was used for quantification of trypsin. The results are compared and the differences between the techniques discussed. Both methods gave similar results, as high or low values in trypsin quantity were reflected in high or low values of tryptic activity. Quantity and activity were linearly and positively correlated, but small differences between methods were found at the lowest detection limits. Both techniques reflect high variability between individual larvae.     

A sensitive method for determining nitrogen-15 isotope in urea

A method is presented in which the ¹⁵N at.% of urea is determined with high precision on liquid samples containing as little as 10 nmol of urea. The method involves removal of interference from NH₄ ⁺ initially present in the sample by cation exchange. Urea in the sample is subsequently hydrolyzed to NH₄ ⁺ by the enzyme urease. Liberated NH₄ ⁺ is separated from the alkaline sample by diffusion as NH₃ through a helium gas phase where it is finally oxidized to N₂ by reaction with hypobromite iodine. The isotopically labeled N₂ thus formed is mixed with the N₂ initially present in the sample, and the ¹⁵N at.% of urea is calculated from the relative amounts of ¹⁴N¹⁵N and ¹⁵N¹⁵N by means of the isotope pairing principle. The presented method for ¹⁵N-urea analysis proved to be precise (SE < 0.4 at.%, n = 5), when applied on both marine and freshwater samples with an ¹⁵N enrichment >1 at.%, and interference was found only from volatile methyl amines. Received: 19 September 1996 / Accepted: 25 October 1996     

Comparison of methods for the analysis of dissolved urea in seawater

A comparison between the diacetyl monoxime and urease methods for measuring dissolved concentrations of urea in seawater was conducted in artificial seawater, phytoplankton-culture filtrate and both natural and ureaspiked field samples from coastal and oceanic enviroments during 1984. The urease technique underestimated urea concentrations in unbuffered photoplankton-culture filtrate as a result of the inhibition of the urease enzyme, causing the incomplete hydrolysis of urea in these samples. Factors responsible for inhibiting urease included pH, seawater ions, and possibly extracellular metabolites produced in unialgal cultures. Seawater type and time of sample collection were important variables affecting urea measurement by the urease method, and recovery of internal standards ranged from 40 to 100%. Increasing the heating time of the urease assay, or the concentration of urease added to the seawater samples increased the amount of urea determined by the urease method. However, measured values were still less than the concentration of the urea internal standards. The diacetyl monoxime method was suitable for urea determinations in all the seawater samples we examined; it was easily automated, and the results were accurate and reproducible. This modified technique is recommended for measuring disolved concentrations of urea in seawater.     

Effects of ambient ammonia levels on blood ammonia, ammonia excretion and heart and scaphognathite rates of Nephrops norvegicus

During commercial handling of Nephropsnorvegicus (L.) there are a number of situations when the prawns may be exposed to very high ambient ammonia levels. These experiments evaluated the effects of increased levels of ambient total ammonia (TA = NH₃ + NH₄ ⁺) on␣blood ammonia, ammonia efflux rates and on the cardio-ventilatory performance of N. norvegicus. When prawns were taken from <1 to 2000 μmol TA l⁻¹ medium, blood TA concentrations increased rapidly for the first 2 h but tended to drop thereafter. Original blood TA levels were restored 6 h after the prawns were transferred back from seawater containing 2000 to <1 μmol TA l⁻¹. Sudden exposure to 500, 1000, 2000 or 4000 μmol TA l⁻¹ medium induced blood TA concentrations to increase respectively to 50, 30, 33 and 36% of external concentrations (normally, internal TA values are much higher than external levels). Immediately after transfer back to seawater with low ammonia concentration ( <1 μmol TA l⁻¹), excretion rates were higher than those of control prawns, and the absolute amounts of TA excreted were considerably higher than those calculated to have accumulated in the haemolymph. Heart rate (HR) and scaphognathite rate (SR) were not altered when prawns were subjected to sudden alterations in ambient ammonia ( <1 to 2000 to <1 μmol TA l⁻¹). When water ammonia concentrations were altered more gradually, both rates increased, but only at 4000 μmol TA l⁻¹. These results show that N. norvegicus is able to remove ammonia from the haemolymph and/or transform ammonia into some other substance when subjected to increased levels of ambient ammonia. Possible mechanisms involved (e.g. active transport across the gills; storage in some other tissue; glutamate synthe sis) are discussed. Received: 20 May 1996 / Accepted: 1 July 1996     

Feasibility of using an automated colorimetric procedure for the determination of seawater nitrate in the 0 to 100 nM range: Examples from field and culture

An automatic colorimetric procedure for measuring nanomolar quantities of nitrate and nitrite in seawater has been improved in sensitivity. Application of this method toin situ investigations in the North Atlantic Ocean in January 1989 and algal culture (Skeletonema costatum) experiments in the laboratory demonstrated that one can record concentrations in the 0 to 100 nM range using classical Technicon AutoAnalyzer equipment. In this working range highest precision obtained was ±2.87 and ±1.24 nM for nitrate and nitrite, respectively.     

Bisphenol A and metabolites released by biodegradation of polycarbonate in seawater

This is the first report of bisphenol A release from polycarbonate during biodegradation by marine microorganisms. Bisphenol A is a monomer in polycarbonate and an endocrine disruptor toxic for marine organisms. Biodegradation of polycarbonate is poorly documented. Here, we have tested the possible release of bisphenol A and metabolites during biodegradation of polycarbonate by marine microorganisms. Polycarbonate degradation was carried out in vitro using a mixed marine microbial consortium isolated from the Bay of Bengal, India, 1 year under controlled laboratory conditions. The degradation was monitored by elemental analysis (EA), Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance spectroscopy (NMR), and gas chromatography—mass spectrometry (GC–MS). The organic soluble metabolites were analyzed by high-performance liquid chromatography (HPLC). We found that bisphenol A was released. The amount of bisphenol A released during 1 year is higher than the half-maximal effective concentration (EC₅₀) values reported for marine organisms. We also identified the following bisphenol A metabolites: 4-hydroxyacetophenone, 4-hydroxybenzaldehyde, and 4-hydroxybenzoic acid. Polycarbonate biodegradation was evidenced by gravimetric weight loss and Fourier transform infrared spectroscopy. The reduction of methyl and carbonyl indices suggests oxidation and hydrolysis of the polymer, respectively. 2D NMR showed an aromatic C–C cleavage.     

A description of a highly diverse soft-bottom community in Kingston Harbour,Jamaica

A highly diverse soft-bottom community in Kingston Harbour, Jamaica is described to provide additional information on the structure of tropical benthic communities. It is shown that the community has a level of diversity unique to stable tropical environments, that dominance by a few species is greatly reduced,and that there are no true parallels to this community in similar environments. The community described seems to fit Sanders' stability-time hypothesis. The density and standing crop of the community are low, and possible explanations for this are discussed. Continuous year-round breeding, short life-spans, and fast turnover rates are suggested as the most important factors regulating biomass in the community described.     

Effect of temperature on seawater desalination-water quality analyses for desalinated seawater for its use as drinking and irrigation water

The effect of feed seawater temperature on the quality of product water in a reverse osmosis process was investigated using typical seawater at Urla Bay, Izmir region, Turkey. The tests were carried out at different feed seawater temperatures (11–23°C) using two RO modules with one membrane element each. A number of variables, including pH, conductivity, total dissolved solids, salinity, rejection percentage of a number of ions (Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl⁻, HCO₃ ⁻, and SO₄ ²⁻), and the levels of boron and turbidities in collected permeates, were measured. The suitability of these permeates as irrigation and drinking water was checked by comparison with water quality standards.     

A technique for collection of particulate organic matter in situ

A technique is described for concentrating and collecting particulate organic matter from water with minimal disturbance. This involves exposing in situ the surface of a Millipore^® filter monitor mounted on a simple vacuum flask assembly.     

Use of tritiated substrates in the study of heterotrophy in seawater

An improved method is described for the study of heterotrophic utilization of dissolved organic substances by marine microorganisms. The method is based on the use of ³H-labelled organic substrates of very high specific activity, rather than the conventionally used ¹⁴C-labelled substrates. Direct measurement of the rate of tracer uptake at near ambient concentration can thus be made instead of extrapolation using the Michaelis-Menten equation. The method also permits comparison between the rates of tracer uptake in sub-samples exposed to different physico-chemical conditions (temperature, light, pollutants, etc.) without the necessity of determining the ambient substrate concentration. The method was applied to the determination of D-glucose uptake by nearshore and pelagic natural microbial populations, and was found to be sensitive and convenient.     

Gonopore sexing technique allows determination of sex ratios and helper composition in eusocial shrimps

An evaluation of the social organization and sexual system of eusocial species of Synalpheus has been hindered because it has not been possible to determine the sexual composition of colony helpers (workers). The external sexual characters typically used to sex caridean shrimps are lacking in Synalpheus. We used SEM sexing technique to determine the sexual composition of helpers in colonies of Synalpheus regalis, S. rathbunae, S. chacei, S. rathbunae A (see Morrison et al. Mol Phylogen Evol 30:563–568, 2004), and S. filidigitus. Colonies consisted of both sexes and sex ratios of helpers generally conformed to 50:50 female to male. Females were characterized by gonopores with U-shaped slits on the coxae of the third pereopods (first walking legs) while males had oval gonopore openings on the coxae of the fifth pereopods (third or last walking legs). In S. chacei, S. filidigitus, and S. rathbunae A, a few helpers were found that had both male and female gonopores (intersexes). All three reproductive females (queens) of S. filidigitus examined were intersexes. Sexing of helpers allowed us to test some hypotheses about sexual differences in helper morphology that might indicate task specialization (division of labor). Male helpers were not different from female ones in body size (except in S. regalis: males somewhat larger) and in fighting chela size. The lack of sexual dimorphism in these characters suggests no male–female specialization in colony tasks such as defense. The presence of male and female helpers similar in size suggests that the sexual system of these eusocial species is gonochoristic, although protandry of some sort in S. filidigitus can not be ruled out. The intersexuality observed in a few individuals may be due to developmental anomalies, protandry, or even simultaneous hermaphroditism. Finally, the sexing technique allowed us to establish that new colonizers of unoccupied sponges in S. rathbunae are a single male and female of helper size.     

A whole-animal bioassay for the determination of the food attractants of the lobster Homarus gammarus

A semi-quantitative assay is described for assessing the effectiveness of bait extracts for the lobster Homarus gammarus (L.), using an extract of the squid Loligo vulgaris Lamarck as a standard. Fractionation of this extract indicated that, while no single component was as attractive to lobsters as the whole, the recombination of the various components yielded a solution which was as attractive as the original extract. It appears likely that several different classes of chemosensory cell must be simultaneously stimulated for the initiation of feeding behaviour in the lobster.