Human population induced eutrophication (pollution) of water sources throughout the developed & developing world is increasing the occurrence of blue-green algal scum (H.A.B.) formation on water bodies and even when eutrophication is removed these occurrences become self sustaining. The number of toxin types identified is increasing & the legislated safety limits for drinking & recreational water is continuously being revised down. A H.A.B. toxins testing service has been set up by us to meet the increasing legislative demand for enforcement & monitoring of the increasing occurrence of toxic blue-green algal presence in water bodies used by humans, commercially important domestic animals and wildlife.

SIGNIFICANT FACTORS IN H.A.B. OCCURENCE
Planktonic blue-green algae concentrate horizontally near the water surface, capturing most of the sunlight that fuels productivity in the aquatic environment. A dense bloom can develop under conditions such as warm, calm weather and when nutrients (phosphorus and nitrogen) are not scarce. It is with the arrival of a breeze that concentration of a scum begins along the leeward shore, resulting in an increasingly toxic dose in a greatly reduced volume. The wide range of toxins produced by cyanobacteria includes hepatotoxic peptides, neurotoxic alkaloids, cytotoxic alkaloids, saxitoxin derivatives, lipopolysaccharides and allergens. Approximately 50% of all cyanobacterial are reported to be highly toxic and when a Microcystis bloom this goes to approximately 80%. These toxins are among the most deadly and include the most potent tumour promoter known to man. Even when not toxic, a dense blue-green algal presence can result in daylight production of high O2 concentrations resulting in oxygen stress and conversely during the night aerobic respiration can result in severe anoxia, which adversely affects aquatic life e.g. fish kills due to asphyxiation. The bloom as it decays will result in a general deterioration of water quality presenting foul odours and water treatment problems

HARMFUL ALGAL BLOOM ANALYSIS SERVICE
We offer a service that enables customers to demonstrate due diligence, properly documented, in regard to the safety of their products and services:

A low cost monitoring service to assist with forward planning & risk assessment.

MONITORING                                                                     PRICE (Excl. V.A.T.)                    *ALERT LEVEL

● Microscopic analysis
Photographed as a permanent record of
species present & cell density within sample                            £10                                           20, 000 cells per ml

● Chlorophyll a Concentration
by Spectroscopic Analysis                                                              £15                                            10 mg chlorophyll a per litre

*WHO recommended trigger level (Chorus & Bartram, 1999), above which actions must be taken e.g. post on-site advisory signs and inform relevant authorities. Below these alert levels there is a relatively low probability of adverse health effects.

If, during monitoring, the bloom density has reached the alert level  (~20, 000 cells per ml &/or 10 mg chlorophyll a per litre) we will (upon request) detect & quantify any H.A.B. toxins that may be present such as the Hepato- & Neuro-toxins which are toxic to liver and to nervous system respectively.

Assay Type                                                                            PRICE (Excl. V.A.T.)            PURPOSE

Biochemical Assays
● Protein Phosphatase Inhibition Assay                                                                             Screening, followed by HPLC
Detection of hepatotoxins. Method calibrated                                                                confirmation if (okadaic acid,
against Lab Reference Material of known                                                                         microcystins and nodularin) detected
hepatotoxicity & a Microcystin-LR calibration curve.                 £20

● Acetylcholinesterase Inhibition Assay                                                                            Screening, Detection of Organophosphate
Detection of Anatoxin-A(s)                                                                 £20                            neurotoxins, Anatoxin-A(s) &/or Man                                                                                                                                                               made Organophosphates. Screening,                                                                                                                                                               followed by HPLC confirmation

Invertebrate Toxicity-Based Bioassays
● Desert Locust (All Cyanotoxins elicit toxicity             Single determination            General toxicity & ecotoxicity
however esp STX LD50 is 8 ng g-1 but not as                   in triplicate £12                 assessment & Specific bioactivity
sensitive to MC-LR or ANTX-a.                                                   LD50 £120                       detection e.g. insecticidal
                                                                                                                                                         &/or anti-protozoal activity
● Brine shrimp                                                                                                                              to detection of unknowns.
Artemia salina (detects toxicity esp. MC & NOD)         Single determination
evaluated against hepatotoxic blooms e.g.                        in triplicate £10
correlated well with mouse i.p. & HPLC.                                 LD50 £105

● fruit fly                                                                                    Single determination
Drosophila melanogaster                                                           in triplicate £12
Insensitive to neurotoxic Aphanizomenon.                              LD50 £120

● Protozoal sp.                                                                        Single determination
                                                                                                          in triplicate £10  
                                                                                                             LD50 £100

ELISA                                                                                          £315 for 42 samples
                                                                                                              £7.50each

Instrumental Analyses
HPLC-DAD                                                                                              £20

HPLC can be linked to any of the other                                                                                Detection & quantification the major
assays described e.g. to screen unknown                                                                            known cyanobacterial toxin classes e.g.
‘peaks’ for inhibition of protein phosphatase                                                                      Microcystin(s), Anatoxin-A, STX class
enzymes that would be indicative of unknown                                                                  Cylindrospermopsin
hepatotoxin(s).
 
●          When the Cyanotoxins present reach the guideline value (e.g. WHO value for Microcystin-LR eq at 1 mg l-1) in e.g. a potable water source, (if requested) we can track the concentration of these problem compounds throughout ‘clean up’ from each stage from source to the finished product (tap or bottled water). See contract research regarding customer project design.

CUSTOMER SAMPLES
Water sample intracellular, extra cellular and total toxin content can be determined if requested and is treated as three separate determinations. These distinctions are unnecessary with filtered water where all present will be ‘free toxin’. No charge for the extra processing.

Upon request and discussion (overlapping with our contract research) we will examine other relevant materials such as e.g. fish (whole, specific tissues, liver, guts) Shellfish, stomach/Rumen contents or equivalent, plant material and other relevant material associated with H.A.Bs.

Depending upon the nature of the sample, there may be additional costs incurred due to processing prior to analysis.

WATERBODY SAMPLING GUIDELINES 

• Collect water sample on leeward shore (i.e. downwind) where algal scum has greatest concentration.
• For monitoring, a 10 ml sample sampled at or near surface is sufficient.
• If algal scum is below the waterline sample at or near surface (500 ml volume minimum recommended e.g. a clean plastic drinks bottle), if above scoop into a labelled container (100 ml volume minimum recommended) and obtain as much concentrated material as container volume allows.
• Labelling information should include: Waterbody name & code (large waterbodies only, obtainable at www.uklakes.net), customer sample identifier, national grid reference, date & time of sampling, contact details
• For large water bodies several selected sites to be sampled
• Record of routine sampling site(s) & extent of algal scum e.g. photograph of waterbody and scum from several recorded viewpoints (optional). Photographically record any suspicious animal deaths as the posture and appearance may give an indication of toxin class present.
• Keep sample(s) as cool & dark as possible. For samples to be analysed within 48 hours store in fridge as soon as possible, if anticipated to be longer prior to analysis then freeze.
  

Miscellaneous Material Sampling Guidelines
Much is dependent on nature of the sample however it is recommended:

• 500 ml volume minimum for a liquid recommended e.g. a clean plastic drinks bottle) and for the solid/semi-solid scoop into a labelled container (100 ml volume minimum recommended) and obtain as much concentrated material as container volume allows.
• Follow as many of the above guidelines as possible and where relevant.
  

Report Timescale
Results are reported back within 48 hours of receipt for water and algal material. Animal, plant & miscellaneous material may require additional processing and may influence report timescale.