No messing about on river conservation!


Scientists from the University of Portsmouth have been investigating nutrient concentrations in the Upper River Itchen, in Hampshire (GB), on behalf of Salmon & Trout Conservation (S&TC) to better understand where phosphorus is coming from and how it is impacting river ecology.

The work has been ongoing for over three years and Lauren Mattingley, Science Officer for S&TC says: “Continuous monitoring of phosphorus has improved our understanding of nutrient dynamics in the Itchen. So far, the results from this monitoring have influenced the lowering of discharge limits from watercress companies and trout breeding farms.

“The behaviour of phosphorus in our rivers is relatively poorly understood, and this is often reflected in water quality standards that, in our opinion, lack the scientific evidence to adequately protect the ecology of the UK’s diverse water resources. Research like that which we have commissioned on the Itchen is essential to set informed phosphorus permits to protect our water life.”

The Itchen is a world famous chalk stream; renowned for its fly fishing and clear water. Designated a ‘Special Area of Conservation’ (SAC) the river supports populations of water-crowfoot, Southern damselfly, Bullhead, Brook lamprey, White-clawed crayfish and otters. The upper river does not suffer from wastewater treatment plant discharges, but does support two watercress farms, which have been the focus of initiatives to reduce phosphate concentrations.

S&TC is the only British charity campaigning for wild fish and their habitats. The organisation’s goal is for British waters to support abundant and sustainable populations of wild fish and all other water-dependent wildlife. Within its ‘Living Rivers’ campaign S&TC is seeking to tackle two of the major causes of poor water quality – fine sediment and phosphorus. The Itchen is therefore being treated as a pilot river for their water quality monitoring initiatives.

Phosphorus in fresh water is a major concern globally; mainly because of its role in the formation of algal blooms and eutrophication, which have a harmful effect on water quality and habitats. Under certain conditions, raised phosphate concentrations contribute to the proliferation of nuisance phytoplankton as well as epiphytic and benthic algae. Diffuse sources of phosphate include storm water and agricultural run-off from land, and point sources include septic tanks and wastewater discharges from industry and sewage treatment works. While soluble reactive phosphorus (SRP) is the main concern, because of its availability for aquatic organism growth, other forms of phosphate such as particulate phosphate can contribute to nutrient enrichment.

Efforts to improve the quality of water bodies in Britain have been underway for many years. The EU’s Water Framework Directive (WFD) required Britain to achieve ‘good status’ of all water bodies (including rivers, streams, lakes, estuaries, coastal waters and groundwater) by 2015, but in 2012 only 36% of water bodies were classified as ‘good’ or better.

In 2013 the UK Technical Advisory Group (UKTAG) published recommendations to revise the standards for phosphorus in rivers, because the standards set in 2009 were not sufficiently stringent – in 75% of rivers with clear ecological impacts of nutrient enrichment, the existing standards produced phosphorus classifications of good or even high status! DEFRA (British Government Department looking after environmental matters), therefore, revised the phosphorus standards to lower concentrations. However, the SRP concentration limits vary widely according to the location and alkalinity of the river.

Recognising a gap in the understanding of the relationship between phosphorus and aquatic ecology, S&TC has a unique agreement with the Environment Agency (EA) in Hampshire in which key environmental targets have been established for the Rivers Test and Itchen to help drive ecological improvements. The agreed targets are set around the number of key water insects that should be expected in a 3-minute kick-sweep sample. The targets are for the middle and lower reaches of the catchment to support at least 10 separate mayfly species and 500 freshwater shrimps (Gammarus) – all of which are susceptible to different forms of pollution so their presence provides an effective measure of the environmental health of the river.

S&TC has also conducted research investigating the effects of fine sediment and SRP on the hatching of the blue winged olive, Serratella ignita (Ephemerellidae: Ephemeroptera) a crucial component of the aquatic food chain. The results found that a cocktail of SRP and fine sediment at concentrations exceeding those found in many UK rivers (25 mg/L fine sediment and 0.07 mg/L phosphate) caused 80% of the eggs in the experiment to die. This work was unique because it showed environmental damage caused by phosphorus beyond eutrophication.

River Itchen sampling and analysis
Five automatic water samplers have been strategically located on the river each collecting daily samples. This generates 120 samples per 24 day cycle, which are collected and transferred to the laboratory in Portsmouth. The samples are split into three for the analysis of Total Phosphate, Soluble Reactive Phosphate (SRP) and Total Dissolved Phosphate (TDP). To cope with such a high volume of work, the laboratory in the University of Portsmouth’s School of Earth & Environmental Sciences employs a QuAAtro 5-channel segmented flow autoanalyzer, from SEAL Analytical.

“The QuAAtro has been in heavy use for over 9 years,” says Senior Scientific Officer Dr Adil Bakir. “It has been employed on a number of academic and commercial research projects, and is also used for teaching purposes. As a 5-channel instrument, we are able to study phosphate, nitrate, nitrite, ammonia and silicate, but our work on the River Itchen is focused on the different forms of phosphate.”

The University of Portsmouth’s Environmental Chemistry Analytical Laboratory provides analytical and consultancy services for businesses, universities and other organisations. Dr Bakir says: “Using the QuaAAttro we are able to analyse diverse matrices including river water, sea water and wastewater, and with automatic dilution and high levels of sensitivity, we are able to measure a wide range of concentrations.”

Creating effective discharge consents
The analytical work undertaken by the laboratory at the University of Portsmouth has greatly improved the understanding of the ecology of the River Itchen and thereby informed the development of appropriate discharge consents for the two watercress farms. Effective 1st January 2016, new discharge permits were issued by the Environment Agency that set limits on phosphate discharges to the River Itchen system. For the Vitacress Pinglestone Farm these limits are set at 0.064 mg/L and are measured as an annual mean increase compared to the inlet sample.

S&TC now works closely with Vitacress, monitoring immediately downstream of the discharge so that the effects of the new discharge limit can be effectively assessed.

Looking forward, Lauren says: “The lessons that we have learned on the Itchen are transferrable, and do not just apply to chalk streams. All rivers have their issues and inputs, so proper diagnosis and understanding of how these inputs shape the biology is essential to the successful restoration of degraded systems.

“In an ideal world phosphorus targets would be bespoke, on a river by river basis, and determined by tailored research and proper monitoring.

“River ecology is impacted by a wide variety of factors and whilst nutrients represent a serious risk, it is important for us to understand all of the threats, and the relationships between them. In summary, without high-resolution monitoring, river standards and river restoration efforts will be blind to their consequences.”

#SealAnalytical #Environmental @SalmonTroutCons @_Enviro_News

Researchers investigate ultra-low Mediterranean nutrient levels.


Researchers at Haifa University’s Marine Biological Station in Israel are exploiting the ultra-low detection limits of advanced laboratory equipment to measure extremely low nutrient concentrations in marine water.

H.Nativ – Morris Kahn Marine Research Station

The University’s Prof. M. D. Krom says: “We work in the Eastern Mediterranean which has the lowest regional concentration of dissolved nutrients anywhere in the global ocean. We therefore utilize an automated segmented flow analyzer from SEAL Analytical, which has been specially adapted to accommodate ultra-low measurements.”

The SEAL AutoAnalyzer 3 (AA3) is a 4 channel system, measuring Phosphate with a long flow cell which has a detection limit of 2 nM. Ammonia is measured using a JASCO fluorometer with a similar ultra-low detection limit, and Silicate, which has a higher concentration, is measured using SEAL’s high resolution colorimetric technology.

The measurement data are being used to determine the season nutrient cycling in the system, which will then be used to help understand the nature of the food web and the effects of global environmental and climate change.

Low nutrient levels in the Mediterranean
The eastern Mediterranean Sea (EMS) has an almost unique water circulation. The surface waters (0-200m) flow into the Mediterranean through the Straits of Gibraltar and from there into the EMS at the Straits of Sicily. As the water flows towards the east it becomes increasingly saline and hence denser. When it reaches the coast of Turkey in winter it also cools and then flows back out of the Mediterranean under the surface waters to Sicily, and then eventually through the Straits of Gibraltar to the North Atlantic. This outflowing layer exists between 200m and 500m depth.

Phytoplankton grow in the surface waters (0-200m) because that is the only layer with sufficient light. This layer receives nutrients from the adjacent land, from rivers and wastewater discharges, and also from aerosols in the atmosphere. These nutrients are utilized by the plankton as they photosynthesize. When the plants die (or are eaten) their remains drop into the lower layer and are jetted out of the EMS. Because the water flows are so fast (it takes just 8 years for the entire surface layers of the EMS to be replaced), these nutrient rich intermediate waters rapidly expel nutrients from the basin. The result is very low nutrient concentrations and very low numbers of phytoplankton – some of the lowest values anywhere in the world. Prof. Krom says: “The maximum levels of nutrients measured in the EMS are 250 nM phosphate, 6 uM nitrate and 6-12 uM silicate. Ammonia is often in the low nanomolar range. By contrast, in the North Atlantic, values are 1000 nM phosphate, 16 uM nitrate and 20 uM silicate, and the levels in the North Pacific are even higher.”

The value of data
The low levels of plankton caused by low nutrient levels, result in a low biomass of fish. Nevertheless, coastal areas generally support more fish than offshore, so the research will seek to quantify and understand the nutrient cycle in the coastal regions, which is poorly understood at present. “We plan to develop understandings which will inform stakeholders such as government. For example, there is a discussion about the potential for fish farms off the Israeli coast, so our work will enable science-based decisions regarding the quantity of fish that the system can support.”

To-date, three data sets have been taken from the EMS, and the first publishable paper is in the process of being prepared.

Choosing the right analyzer
Prof. Krom says that his first ‘real’ job was working for the (then) Water Research Centre at Medmenham in Britain, where he was involved in the development of chemical applications for the Technicon AA-II autoanalyzers, which included going on secondment to Technicon for several months. SEAL Analytical now own and manufacture the AutoAnalyzer brand of Continuous Segmented Flow Analyzers, so his career has been connected with autoanalyzers for decades. For example his is Professor (Emeritus) at the University of Leeds (GB), where, again, he worked with SEAL autanalyzers. An AA3 instrument was employed at Leeds in a project to investigate the nature of atmospheric acid processing of mineral dusts in supplying bioavailable phosphorus to the oceans.

Explaining the reasoning behind the purchase of a new AA3 at Haifa University, Prof. Krom says: “During a research cruise, it is necessary to analyse samples within a day to avoid changes in concentration due to preservation procedures.

“Typically we analyse 50-80 samples per day, so it is useful to useful to be able to analyze large numbers of samples automatically. However, the main reasons for choosing the SEAL AA3 were the precision, accuracy and low limits of detection that it provides.”

Commenting on this application for SEAL’s analyzers, company President Stuart Smith says: “Many of our customers analyze nutrient levels in freshwater and marine water samples, where high levels of nutrients are a concern because of increasing levels of algal blooms and eutrophication. However, Prof. Krom’s work is very interesting because, in contrast, he is looking at extremely low levels, so it is very gratifying that our instruments are able to operate at both ends of the nutrient concentration spectrum.

• Powley, H.R., Krom, M.D., and Van Cappellen, P. (2017) Understanding the unique biogeochemistry of the Mediterranean Sea: Insights from a coupled phosphorus and nitrogen model. Global Biogeochemical Cycles, 11; 1010-1031. DOI 10.1002/2017GB005648.

• Stockdale, A. Krom, M. D., Mortimer, R.J.G., Benning, L.G., Carslaw, K.S., Herbert, R.J., Shi, Z., Myriokefalitakis, S., Kanakidou, M., and Nenes, A., (2016) Understanding the nature of atmospheric acid processing of mineral dusts in supplying bioavailable phosphorus to the oceans. PNAS vol. 113 no. 51

#SealAnal #Marine @_Enviro_News

Discrete or Continuous Flow Analysis – which is better?


A wide variety of factors affect the choice of analytical instrument. These include target workload (samples/hour), variety of chemistries, methods required, bench space, staff availability etc. In the following article Lalicia Potter, Technical Sales & Support Director at SEAL Analytical, examines one of the common decisions facing laboratory managers.

As the manufacturer of an instrumentation range that includes both discrete analyzers and continuous segmented flow analyzers, SEAL Analytical’s technical support chemists are often asked which is the better technique. Both offer fast, automated, colorimetric analysis of multiple samples, however, the answer depends on the current and future analytical requirements of the laboratory.

Descrete Analyser

Descrete Analyser

SEAL’s discrete analysers employ sample trays and discrete reaction wells in which the colorimetric reaction takes place. In contrast, segmented flow analysers (SFA) employ a continuous flow of samples and reagent, segregated by air bubbles within tubing and mixing coils.

In general terms, discrete analysers are ideal when automation is a priority and/or when many and varied tests are needed on different samples. SFA is ideal when a larger number of samples are to be analysed for a smaller number of chemistries. However, both techniques are flexible, so it is important that expert advice is sought in the choice of analyzer and that the instrument is configured to meet the precise needs of the laboratory.

Discrete Analysers
In order to minimise operator involvement, SEAL’s discrete analyzers are highly automated and simple to set up and run, even overnight. A robotic sampling arm works in conjunction with a stepper motor-driven syringe that is responsible for aspirating, dispensing and mixing accurate and precise quantities of sample and reagent. The SEAL AQ1 and AQ2 discrete analyzers can run seven different chemistries from each sample in the same run – and another seven in another run. These instruments have three separate wash stations including a unique probe washer, so cross-contamination is not a problem. This unique washing feature means that even ammonia (using Phenate), nitrate by cadmium reduction– (using ammonium chloride buffer) and low level phenol can be run together with no issues.

SEAL has also built an auto-dilution feature into the discrete analysers for preparing standards automatically and handling over-range samples. These diluted sample results are automatically bracketed by QC sets.

The reproducibility and detection limits of these discrete analysers have been optimised by ensuring that each sample is read in the same optical glass cuvette with a 10mm path length. The sample is always read in the same position in front of the detector, which eliminates any potential issues with scratching or reaction well variability that can be found with direct-read systems. Since the liquid is moved and not the tray; fewer moving
parts maximises reliability.

Most discrete analysers employ miniaturised components to reduce reagent consumption and waste costs. For example, both the AQ1 and AQ2 analysers use just 20 to 400µl of reagent per sample.

Segmented Flow Autoanalysers
Based on the original tried and tested technology of the Technicon™ /Bran Luebbe™ AutoAnalyzer, today’s SFAs deliver fast, accurate analysis for enormous numbers of samples; the QuAAtro for example can run up to 600 tests per hour. SFA’s are also highly automated and once the analyzer is configured and the reagents and samples are loaded, reliable unattended operation is a major benefit.

Flow Analyser

Flow Analyser

A basic SFA system consists of an autosampler, a peristaltic pump, a chemistry manifold, a detector and AACE data acquisition software. Sample and reagents are pumped continuously through the chemistry manifold and
air bubbles are introduced at precisely defined intervals, forming unique reaction segments which are mixed using glass coils. With SFA, even slow reactions run to completion and the ratio of sample to reagents in the detector reaches a constant maximum value; the steady-state condition.

SFAs have been developed for running a few parameters on a larger number of samples, and the SEAL SFAs are the system of choice for marine and seawater organisations and anyone running very low nutrient waters. The SEAL AutoAnalyzer 3 and QuAAtro deliver high levels of performance and reproducibility, and are also the systems of choice for tobacco, soil and fertiliser testing around the world. These analysers provide maximum sensitivity by ensuring that the reaction always goes to completion, and with a digital true dual-beam detection system with real time referencing, the highest reproducibility and very lowest detection limits are achieved.

In summary, when choosing the most appropriate analytical technique, it is important to consider both the current and likely future needs of the laboratory. However, one of the reasons behind the large numbers of SEAL instruments in laboratories around the globe, is that each analyzer has been configured to meet the individual needs of its laboratory. So, it is good practice to contact SEAL’s technical support team at an early stage because if the question is: “Which technique is better,” the answer is: “It depends…”