Viability assessed with the most probable number dilution culture method after chemical treatment of ballast water reveals the presence of false negatives from an approved vital stain method.

The present study compares the CMFDA/FDA + motility- and the Most Probable Number (MPN) Dilution Culture + Motility methods for testing the viability of ≥10-<50 µm organisms in chlorine treated ballast water. The results of both methods were within the regulatory compliance criterion <10 organisms/mL, but the MPN-method revealed that growth-outs did occur. While the CMFDA/FDA method showed <0.5 organisms/mL, the MPN-method gave approx. 6 organisms/mL. This demonstrated that false negatives, i.e. living but not stained organisms, may occur when using the CMFDA/FDA-method for compliance testing of chemical treated ballast water. Organisms surviving the treatment were primarily the dinoflagellate Scrippsiella sp. and various coccoid chlorophytes present in a brackish- and freshwater test, respectively. It is suggested that their resilience to the chemical treatment is the ability to transform into a temporary cyst (Scrippsiella sp.) or the presence of a chemical resistant cell wall (certain chlorophytes).

Inherent lack of CMFDA/FDA staining in certain algae and its implication for ballast water testing.

The approved method for testing the efficacy of ballast water management systems with respect to killing 10-50 µm organisms uses movements of the organisms or the vital stains CMFDA/FDA. The present study demonstrates that certain freshwater coccoid chlorophytes, known or suspected to contain a highly resistant cell wall component (algaenan), stain poorly with CMFDA/FDA, resulting in false negatives. The staining rates for the most dominant species were determined and were approx. 3-70 %. The use of Crystal Violet as an indicator for the presence of algaenan gave inconclusive results. The number of the 10-50 µm organisms in a small pond was found to be 10,183 organisms/mL (Lugol's fixed sample) vs. 2335 organisms/mL (CMFDA/FDA-stained sample). Using the staining rates obtained, it was estimated that the number of false negatives could make 40-50 %. The implications for biological performance evaluation of ballast water management systems are discussed.

Ballast water treatment and bacteria: Analysis of bacterial activity and diversity after treatment of simulated ballast water by electrochlorination and UV exposure.

Effects of ballast water (BW) treatment by ultra-violet (UV) light and electrochlorination (EC) on survival, activity and diversity of marine bacterioplankton and release of organic matter from cell damage were examined at discharge in a large-scale BW test facility (250 m3 tanks) at Hundested harbour, Denmark. The tests were performed in accordance with the requirements for type approval testing by International Maritime Organization (IMO) and US Coast Guard. After treatment, the water was held in the tanks for one day (EC) before discharge, or 6 days (UV, including also a final UV re-treatment) before discharge. In the discharged and treated water, numbers of viable bacteria and bacterial growth rate had decreased significantly relative to the untreated water, but the total number of bacteria only was reduced in the EC-treated water. After additional storage for up to 10 days in small-scale laboratory incubations, significant regrowth of bacteria was observed after either treatment. Sequencing of 16S rRNA gene amplicons demonstrated that α-Proteobacteria initially were dominant, but γ-Proteobacteria dominated after regrowth. Bacteria used to document BW treatment efficiency (E. coli, Vibrio spp., enterococci) survived both treatments; neither treatment reduced the risk of pathogen dispersal. Concentrations of amino acids in the water were used as indicators of treatment-induced cell damage and demonstrated higher concentrations at discharge, but only after the EC treatments. Our results indicate that activity of bacteria, rather than their abundances, should be used when examining effects by ballast water treatment on microorganisms and that none of the examined treatment technologies could eliminate pathogenic bacteria.

Environmental persistence of organic pollutants: guidance for development and review of POP risk profiles.

Environmental persistence is an important property that can enhance the potential of a chemical substance to exert adverse effects and be transported to remote environments. The persistence of organic compounds is governed by the rates at which they are removed by biological and chemical processes, such as biodegradation, hydrolysis, atmospheric oxidation, and photolysis. The persistence workgroup in a recent Society of Environmental Toxicology and Chemistry (SETAC) Pellston workshop (Pensacola, FL, USA, January 2008) focused on evaluating persistence of organic compounds in environmental media (air, water, soil, sediment) in terms of their single-medium degradation half-lives. The primary aim was to provide guidance to authors and reviewers of chemical dossiers for persistent organic pollutants (POPs) and persistent, bioaccumulative, and toxic substances (PBTs) proposed for action. A second objective was to provide a summary of the current state of the science with respect to POP fate assessment. Assessing the persistence of chemical substances in the environment is not straightforward. A common misconception is that, like many chemical properties, environmental persistence is an inherent property of the substance and can be readily measured. In fact, rates of degradation of a substance in the environment are determined by a combination of substance-specific properties and environmental conditions. This article addresses how persistence can be evaluated based on an assortment of supporting information. Special attention is given to several critical issues, including transformation products, nonextractable residues, and treatment of uncertainty and conflicting data as part of a weight-of-evidence assessment.

The effect of short-term hyperammonaemia on milk synthesis in dairy cows.

To test the hypothesis that ammonia detoxification in ruminants consumes amino acids to the detriment of milk protein production, we infused four lactating dairy cows with ammonium acetate or sodium acetate in switchback experiments. Plasma ammonia concentrations increased to 411 microm within 1 h of the start of infusion of ammonium acetate at 567 mmol/h. The rate constant for ammonia clearance from plasma was 0 x 054/min and the half-life was 12 x 9 min. Infusion at 567 mmol/h for 1 h followed by 1 h without infusion, repeated four times between am- and pm-milking, caused a decrease in feed intake. Compared with sodium acetate, continuous infusion of ammonium acetate at 360 mmol/h throughout an entire 10-h milking interval increased plasma ammonia concentrations to 193 microm and caused a 20% decrease in milk, protein and lactose production with no effect on percentage composition of milk or the yield of milk fat. Arterial concentrations of glucose and non-esterified fatty acids tended to increase; there was no effect on arterial acetate, beta-hydroxybutyrate or triacylglcerol, and branched-chain amino acids, Lys and Thr decreased. Mammary plasma flow, estimated by assuming 100% uptake/output of Phe+Tyr, was significantly correlated with milk yield. Mammary uptakes of acetate tended to be reduced by hyperammonaemia, but uptakes of other energy metabolites and amino acids were not affected. Thus, while an increase in amino acid consumption during hyperammonaemia was apparent from the drop in circulating concentrations of Leu, Ile, Val, Lys and Thr, there was no evidence to support the hypothesis that milk yield is affected by the lower concentrations. An ammonia-induced depression in feed intake may have caused the decrease in milk synthesis.

Comparison of biodegradation of surfactants in soils and sludge-soil mixtures by use of 14C-labelled compounds and automated respirometry.

The biodegradability of dodecyl benzene sulphonate (LAS), nonylphenol-di-ethoxylate (NP2EO) and tridecyl-tetra-ethoxylate (LAE) in soil was examined by use of 14C experiments at two concentrations (10 and 400 mg/kg). Increasing the concentration of test chemical from 10 to 400 mg/kg resulted in a decrease in the relative maximum mineralization rate and an increase in the estimated lag times of a factor of approximately 3.5. In sludge-amended soil, the highest expected environmental concentration (just after sludge application) will be around 10 mg/kg for linear alkylbenzene sulphonate (LAS), while the concentration of NP2EO and linear alcohol ethoxylates (LAE) will be much lower. However, when using a respirometric method it is necessary to use a higher concentration of test substance in order to detect biodegradation. In our experiment, amendment with anaerobically digested sludge resulted in a decrease in the mineralization of LAS, NP2EO and LAE for all soils. Respirometric experiments were carried out at 400 mg/kg and could be used for estimation of biodegradation potential of LAS, NP2EO and LAE in soil and sludge-amended soil. For LAS, the results obtained from the respirometric experiments were similar to the results obtained in the 14C experiments, whereas NP2EO and LAE showed a faster degradation in the respirometric experiments.