BI MONTHLY TOXICITY ANALYSIS OF EWER EMISSIONS AT THE ... · Toxicity testing was requested by for the final effluent sample at the Henkel facility as part of their IPC licensing

This report shall not be reproduced except in full, without the approval of BNM Environmental. All queries concerning the report or its contents

should be forwarded to the Monitoring Team Leader. ______________________________________________________________________________________________________

Bord na Móna Environmental Ltd

Registered Office: Main Street, Newbridge, Co Kildare, Ireland. Registered No: 303312, VAT No: IE6323313B

For the Attention of: Mr. Thomas Lavin

Henkel (Ireland) Ltd.

Tallaght Business Park

Whitestown, Tallaght

Dublin 24

Field work completed by: Mr. Stephen Stapleton

Prepared by: Mr. Stephen Stapleton

Environmental Scientist

Reviewed by: Mr. Peter Coogan

Environmental Team Leader

BNM File Ref: ECS5071-BF-Tox

Monitoring Date: 25th August- Forth Bi-Monthly Event 2015

Monitoring Period: Toxicity 4 of 6 2015

Reporting Date: 9st November 2015

BI-MONTHLY TOXICITY ANALYSIS OF SEWER

EMISSIONS AT THE HENKEL (IRELAND) LTD.

FACILITY IN KYLEMORE PARK NORTH,

BALLYFERMOT, DUBLIN 10 IN COMPLIANCE

WITH THE REQUIREMENTS OF INTEGRATED

POLLUTION PREVENTION AND CONTROL

LICENCE REGISTER: NO. P0078-01.

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Report No. ECS5071-BF-Tox

Henkel (Ireland) Ltd., Ballyfermot Page 2 P0078-01

Executive Summary

Henkel (Ireland) Ltd. is required to assess the toxicity of emissions to water (wastewater discharges)

from the Ballyfermot facility on a bi-monthly basis, to comply with the requirements of the Integrated

Pollution Prevention and Control Licence, Register No. P0078-01. BNM Environmental was contracted

to perform the sampling. A BNM Environmental Scientist visited the Ballyfermot site on the 25th of

August 2015 to carry out the forth bi-monthly monitoring event of 2015.

A sample was obtained from emission reference point BFW-1A for monitoring of the bi-monthly

parameters. The wastewater sample was analysed by Aquatic Services Unit for 48 hour LC50 (Tisbe

battagliai - Marine copepod) and 30 min EC50 (Vibrio fischeri - Microtox).

The 48h LC50 to Tisbe battagliai test shows a toxicity value of 1.2 Toxic Units. The 30min EC50 to Vibrio

fischeri test shows a toxicity value of 5.9 Toxic Units.

This would indicate that the BFW-1A wastewater emission point did register a minor acutely toxic effect

on the test species.

This report is certified as accurate and representative of the sampling and associated analysis carried out.

Respectively Submitted,

__________________________

Mr. Stephen Stapleton Mr. Peter Coogan

Environmental Scientist Environmental Team Leader

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1.0 METHODOLOGIES

1.1 Sampling

Representative grab samples of wastewater were extracted from Hach Lange autosampler in

accordance with the following standards;

TABLE 1.1: STANDARDS

ISO Standard

Description

ISO 5667-1-2006

Guidance on the design of sampling programmes and sampling

techniques

ISO 5667-3-2012

Guidance on sample preservation and handling

ISO 5667-14-1998

Guidance on quality assurance of environmental sampling & handling

ISO 5667-10-1992

Guidance on sampling wastewaters

Sampling was conducted in strict accordance with BNM Environmental recognised Standard

Operations Procedures (SOP) TS-W-02. All samples were returned to the laboratory, and stored

between 1-8°C. The sample was subsequently transported to the Aquatic services unit

Laboratory in Cork, Co. Cork.

1.2 Analysis

Analysis was conducted by Aquatic Services Unit. The wastewater sample was analysed for 48

hour LC50 (Tisbe battagliai - Marine copepod) and 30 min EC50 (Vibrio fischeri - Microtox). Table

1.2 presents the methodology used for the toxicity analysis of the wastewater sample.

TABLE 1.2: TOXICITY TESTING OF WASTEWATER SAMPLE

Test Species Method

Tisbe battagliai The direct toxicity assessment of aqueous environmental samples using

the marine copepod Tisbe battagliai lethality test based on the Environmental Agency 2007 test method

Vibrio fischeri Method 6.2 “Determination of the inhibitory effect of water samples on the

light emission of Vibrio fischeri (Luminescent bacteria test) – Part 3: Method using freeze-dried bacteria” based on ISO 11348-3: 2007

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2.0 RESULTS

The results of the toxicity analysis carried out by Aquatic Services Unit are presented in Table

2.1 as follows:

TABLE 2.1: TOXICITY RESULTS FOR EMISSIONS TO SEWER (BFW-1A)

Test Species Test Parameter Test ResultNote1 No. of Toxic Units

Vibrio fischeri 30 min. EC50 16.9% 5.9

Tisbe battagliai 48 hr. LC50 81% 1.2

Note 1: Effluent Concentration

3.0 COMMENT

The results of the bi-monthly toxicity testing of the wastewater sample extracted from the

emission reference point BFW-1A at the Henkel (Ireland) Ltd. Ballyfermot facility on the 25th of

August 2015 are presented in Table 2.1 above.

The toxicological effect of BFW-1A on the test species has increased when compared with

previous results obtained in July (ECS5071).The 30 min EC50 to Vibrio fischeri has decreased

since the previous monitoring event with a toxicity value 10.7.→5.9 Toxic Units at a concentration

of 16.9%vol/vol.

The 48h LC50 to Tisbe battagliai test has decreased since the previous monitoring event with a

toxicity value of 16.5→1.2 Toxic Units at a concentration of 85%vol/vol.

Based on the results from testing carried out on BFW-1A it would indicated that the samples is

minor acutely toxic.

Please see Appendix 1 for map of monitoring locations

Please see Appendix 2 for Aquatic services unit toxicity report

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Appendix 1: Site Location Map

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Appendix 2: Aquatic services unit toxicity report

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Report on the IPPC Toxicity Testing of the

Henkel BFW-1A final effluent

on behalf of

Anua Environmental Technical Services

September 2015

Report prepared by

Aquatic Services Unit,

Environmental Research Institute. Lee Road.

Cork

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2

Introduction

Toxicity testing was requested by for the final effluent sample at the Henkel facility as part of their IPC licensing requirements. The sample code for the Henkel effluent is BFW-1A and the EPA IPPC License number for the effluent is P0078-1. Two aquatic species from different trophic levels were selected for the testing namely the marine copepod (Tisbe battagliai) and the bacteria (Vibrio fischeri). Testing commenced on August 26th, 2015 and finished on August 28th, 2015.

Methodology

The effluent sample was collected on August 25th, 2015 by Anua Environmental personnel and then transported by courier service to the Aquatic Services Unit (ASU) laboratory. The effluent sample arrived to the ASU laboratory on July 22nd, 2015 and had temperatures of 13.3ºC on arrival; the sample was then immediately placed in a refrigerator at 4 ºC until required for testing. Tisbe battagliai Bioassay

The Tisbe bioassay was carried out following standard methods as described in Environment Agency’s Methods for the Examination of Waters and Associated Materials (Environment Agency, 2007). The effluent was tested for toxicity at the following concentrations 4.5, 9, 18, 36, and 72% effluent. The salinity of the effluent was adjusted to 30 parts per thousand (ppt) using hypersaline brine to make the sample suitable for testing. Twenty Tisbe isolates (animals 6 days old) were tested for each concentration. These animals were added into 4 replicates of five animals per test chamber for each concentration. Testing was carried out in a constant temperature room at a temperature of 20 ºC ± 2 throughout the test. A light regime of 16 hours light/8 hours dark was used throughout the testing period. The test duration was 48 hours. Filtered seawater adjusted to 30 ppt by the addition of distilled water was used as control and dilution water. An additional brine control comprising of distilled water and hypersaline brine was also tested. A concurrent reference toxicant using zinc sulphate was conducted to assess the sensitivity of the test organisms. The T. battagliai were obtained from in house cultures at the ASU.

Vibrio fischeri bacteria bioassay using Microtox system.

The luminescent bacteria Vibrio fischeri are used exclusively in the Microtox system. Testing was carried out following the ISO 11348-3 guidelines (ISO, 2007). Two replicates were used for each concentration tested. The effluent sample was diluted by a factor of 2 using distilled water prior to testing based on the previous evidence of toxicity associated with the effluent. The concentrations of effluent tested ranged from 3.1 % to 40 %. A concurrent reference toxicant bioassay was also carried out to determine the health and suitability of the bacteria.

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Statistical Analyses

Statistical analyses to generate LC50 (Lethal Concentration to cause 50 percent mortality) or EC50 (Effective Concentration to cause 50 percent effect) data were performed using the ToxCalc v5.0.32 Environmental Toxicity Data Analysis System, (Tidepool Scientific, 2007). Microtox statistical analyses were carried out using the manufacturers software programme MicrotoxOmni (Azur Environmental Ltd., 1995).

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Results and Discussion

Tisbe battagliai Bioassays

All of the tests were deemed to be valid given that there was greater than 90% survival in the controls indicating that the animals and testing conditions were satisfactory (Environment Agency, 2007). Survival across the concentration series of effluent ranged from 100% at lowest effluent concentration to 60% at highest concentration tested. Significant toxicity was observed between the controls and the 72% effluent concentration. This bioassay predicted an LC50 value of 81 % effluent. This corresponds to a Toxicity Unit (TU) value of 1.2 for the effluent. Results of this bioassay are displayed in Figure 1.

Figure 1 Average survival of Tisbe battagliai after 48 hours in a concentration series of the Loctite BFW-1A effluent.

A concurrent reference toxicant bioassay using zinc sulphate was carried out alongside the effluent bioassays. This bioassay produced an EC50 of 0.423 mg/L of Zinc sulphate, this EC50 value is in agreement with previously published data for Tisbe

sp. (US EPA, 2007) and with ongoing toxicity testing at the ASU. This indicates that the animals were of suitable sensitivity to be used for toxicity testing. Water quality measurements in the test chambers remained within normal limits for the duration of the bioassay. These data are presented in Appendix 1.

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Microtox results.

The Microtox bioassays carried out on August 27th, 2015 were deemed to be valid. A reference toxicant test was carried out before actual testing of samples took place to ensure that the bacteria and reagents were suitable for testing. The reference toxicant test using zinc sulphate determined an EC50 of 8.5 mg/L after 15 minutes exposure. This is within the guideline range of 3-10 mg/L as specified by the Microtox manufacturer (Azur Environmental, 1995). These data are presented in Appendix 2.

A dose dependent increase in toxicity was observed between the controls and throughout the effluent concentration series. Significant toxicity was observed for effluent concentrations greater than 6.25% effluent. A GL value of greater 16 was generated for this test. The GL value is the dilution level at which an effluent causes less than 20% inhibition to the species. Effluent samples with GL values ranging between 1-10 are considered to have low toxicity, GL values of 10-100 are considered to be moderate toxicity and GL values greater than 100 are considered to have high toxicity. This GL ranking system is based on research carried out on selected effluents. (Wang et al., 2002). This bioassay produced an EC50 of 16.9%. This would equate to a TU value of 5.9 for the BFW-1A effluent. The data for this test are presented in Figure 2.

Figure 2 Plot of the Microtox results for the Henkel BFW-1A effluent sample. Values represent light production relative to the controls. The highest concentration of effluent tested was 40%.

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Summary and Conclusions

All of the bioassays performed in the current round of testing were deemed to be valid as they meet all of the criteria as specified in the guidelines.

Exposure of the marine copepod, Tisbe battagliai, to the Loctite BFW-1A effluent did produce significant toxicity only at the highest concentration tested. The Tisbe bioassay determined a TU value of 1.2.

In the Microtox bioassay, exposure to the BFW-1A effluent did have a statistically significant inhibitory effect on light production in the Vibrio fischeri bacteria. A dose dependent decrease in light production was observed across the concentration series. The Microtox bioassay was more sensitive than the Tisbe bioassay to the effluent and a TU value of 5.9 was determined during this round of testing.

The toxicity of effluents has been classified based on their TU values using the following scale TU<1 (relatively not acutely toxic), TU between 1 and 10 (minor acutely toxic), TU between 10 and 100 (moderately acutely toxic) and TU>100 (very acutely toxic) (Tonkes et al., 1999). Based on this scale, the results from the bioassays performed on the BFW-1A final effluent sampled on August 25th, 2015 indicate that the sample is minor acutely toxic.

References Azur Environmental, 1995. Microtox Acute Basic Test Procedures. Azur Environmental ,. Carlsbad, California, USA. Environment Agency, 2007. The direct toxicity assessment of aqueous environmental samples using the marine copepod Tisbe battagliai lethality test. Methods for the Examination of Waters and Associated Materials. http://www.environment-agency.gov.uk/nls ISO 11348-3:2007. Water quality-Determination of the inhibitory effect of water samples on the light emission of Vibrio fischeri (Luminescent bacteria test)—Part 3: method using freeze dried bacteria. International Standards Organisation. Tonkes, M. Graaf, P.J.F., and Jannes, G.,1999. Assessment of Complex Industrial Effluents in the Netherlands using a Whole Effluent Toxicity (or WET) Approach. Water Science and Technology, Vol. 39, No. 10-11, pp 55-61. U.S EPA, 2007. ECOTOX User Guide: ECOTOXicology Database System. Version 4. http:/www.epa.gov/ecotox. Wang, C., Yediler, A., Lienert, D., Wng, Z., and Kettrup, A., 2002. Toxicity evaluation of reactive dyestuffs , auxiliaries and selected effluents in the textile finishing industry to luminescent bacteria Vibrio fischeri. Chemosphere, 46, pp 339- 344 ..

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http://www.environment-agency.gov.uk/nls

http://www.environment-agency.gov.uk/nls

7

Appendix 1- Water Quality Measurements for Tisbe battagliai Bioassay

48 Hour Tisbe Bioassay

Anua BFW-1 Effluent 26/08/15

Concentration Survival D.O (mg/L) pH Salinity (ppt) Temp (ºC)

(%) 0hrs 24hrs 48hrs 0hrs 24hrs 48hrs 0hrs 24hrs 48hrs 0hrs 24hrs 48hrs 0hrs 24hrs 48hrs

Control A 5 5 5 6.8 6.1 8.00 7.89 30 30 19.1 21.3 21.7Control B 5 5 5Control C 5 5 5Control D 5 5 5Control 2A 5 5 5 6.8 6.1 8.00 7.89 30 30 19.1 21.3 21.7Control 2B 5 5 5Control 2C 5 5 5Control 2D 5 5 5

Brine Control A 5 5 5 6.5 6.5 8.06 7.65 30 31 19.3 21.3 21.7Brine Control B 5 5 5Brine Control C 5 5 5Brine Control D 5 5 5Brine Control 2A 5 5 5 6.6 6.5 8.06 7.65 30 31 19.3 21.3 21.7Brine Control 2B 5 5 5Brine Control 2C 5 5 5Brine Control 2D 5 5 5

4.5 A 5 5 5 6.6 5.8 8.01 7.85 29 30 19.2 21.3 21.74.5 B 5 5 54.5 C 5 5 54.5 D 5 5 5

9A 5 5 5 6.5 6.0 7.96 7.86 29 30 19.2 21.3 21.79 B 5 5 59 C 5 5 59 D 5 5 518 A 5 5 5 6.1 5.8 7.82 7.82 29 29 19.3 21.3 21.718 B 5 5 518 C 5 5 518 D 5 5 536 A 5 5 3 5.9 5.1 7.99 7.66 29 29 19.2 21.3 21.736 B 5 3 336 C 5 4 536 D 5 5 572 A 5 5 3 5.3 1.2 8.12 7.26 29 30 19.0 21.3 21.772 B 5 4 472 C 5 4 172 D 5 4 4

Notes Adjusted pH of 36 and 72% from 7.47 to 7.99 and 6.87 to 8.12 respectively. Aerated 72% from 3.2 mg/l to 5.3mg/l prior to initiation.

Day 0 Initiated at 18.00 , Animals 6 days old at initiation

24hrs 17.00

48hrs Terminated at 16.35

Testing performed by Aquatic Services Unit, ERI Building, Lee Rd., Cork

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Appendix 2- Microtox Reference Toxicant Bioassay

Plot of light inhibition relative to the controls versus zinc sulphate concentration

after 5 and 15 minutes exposure.

-20

0

20

40

60

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100

0 20 40 60 80 100

Lig

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Inh

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(%

)

Concentration (mg/L)

Microtox Zinc Sulphate Bioassay

5 Min Data

15 Min Data

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The toxicity testing carried out for this report complies with internationally accepted guidelines. The results of these toxicity tests contained in this report are deemed valid under these guidelines. Signed:

David Gillespie MSc (Toxicology)

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Documents

BI MONTHLY TOXICITY ANALYSIS OF EWER EMISSIONS AT THE ... · Toxicity testing was requested by for the final effluent sample at the Henkel facility as part of their IPC licensing