A Training Manual for Assessing Pollution

(Supported by FAO-BoBLME Project)

A Training Manual for Assessing Pollution (Trace/Heavy metals) in Rivers, Estuaries and Coastal Waters-Using Innovative Artificial Mussel (AM) Technology - Bangladesh Model

Md Maruf Hossain Golam Kibria Dayanthi Nugegoda TC Lau Rudolf Wu

January 2015 0


A Training Manual for Assessing Pollution (trace/heavy metals) in Rivers, Estuaries and Coastal waters-Using Innovative “Artificial Mussel (AM) Technology” - Bangladesh Model.

By Professor Md M.Maruf Hossain Institute of Marine Sciences and Fisheries (IMSF), University of Chittagong, Bangladesh Dr Golam Kibria and Prof. Dayanthi Nugegoda School of Applied Sciences, RMIT University, Australia Chair Professor T.C. Lau Department of Biology and Chemistry, City University of Hong Kong, Hong Kong Chair Professor and Director Rudolf Wu School of Biological Sciences, The University of Hong Kong, Hong Kong

Citation Hossain, M. Maruf., Kibria, G, Nugegoda, D, Lau, TC and Wu, R. 2015. A Training Manual for Assessing Pollution (trace/heavy metals) in Rivers, Estuaries and Coastal waters-Using Innovative “Artificial Mussel (AM) Technology” Bangladesh Model. Research collaboration between scientists of the IMSF, University of Chittagong, Bangladesh, RMIT University, Australia, the City University of Hong Kong, and the University of Hong Kong. 24p. DOI: 10.13140/RG.2.1.4384.4644 https://www.researchgate.net/publication/273775181_A_Training_Manual_for_Assessing_Pollution_%28traceheavy_metals%29_in_Rivers_Estuaries_and_Coastal_ waters-Using_Innovative_Artificial_Mussel_%28AM%29_Technology_-_Bangladesh_Model

Correspondence about the technical report [email protected] [email protected]

Cover layout: Dr Golam Kibria General disclaimer The opinions, presentation of material and positions expressed in this publication are those of the authors and do not necessarily represent the official views of University of Chittagong or of the supporting agencies.

Financial support by FAO under the project on “Sustainable Management of the Bay of Bengal Large Marine Ecosystem (BoBLME) Project” (Supported by GEF) on Pollution Monitoring in Bangladesh (Artificial Mussel Technology) FAOBGDLOA2014-031.

Acknowledgements Special acknowledgment to Dr Chris O’Brien, Regional Coordinator, Dr Rudolf Hermes and Mr C. L. Andreasson of BoBLME for their support and valuable inputs throughout the program. We acknowledge Prof. S.H.Chowdhury (former Professor and Chairman, Department of Zoology, University of Chittagong) for reviewing the booklet. We sincerely acknowledge the personal and professional efforts of Prof TC Lau and Post-Doc Researcher Dr Ruwei Wang at City University of Hong Kong whose painstaking analysis of all the AM samples and some LM samples made it possible to produce this technical document. Debbrota Mallick (Deb), provided significant support in this collaborative international research project both as an M.Sc research student of the Institute of Marine Sciences and Fisheries (IMSF), Chittagong University, Bangladesh (where he was guided and supervised by Professor M Hossain and Dr Golam Kibria respectively for his M.Sc thesis) and later as a research officer of this project (phase-2). Deb provided valuable field support in deployment and retrieval of AM in Chittagong, Cox’s Bazar, Dhaka and Khulna, collected and collated field information as and when required and helped in producing some maps and diagrams. Other researchers in this research programme (phase-II) are Md. Didarul Alam and Hillol Barua, as part of their B.Sc (Hons.) research work (term paper) who contributed significantly during deployment and retrieval of AM in Chittagong coast, Dhaka and Khulna sites. Md. Tariqul Islam (former IMSF student & now Scientific Officer in NORI- National Oceanographic Research Institution, Cox’s Bazar) provided helpful support during uptake of metals test by AM and LM (Perna viridis) at Cox’s Bazar sites and Mosheskhali sites.

Feedback/comments

We would appreciate to receive any feedback or comments about this technical report. Please send your comments or feedback to the following scientists. Dr Golam: [email protected] Prof. Hossain: [email protected]

1


A Training Manual for Assessing Pollution (Trace/Heavy Metals) in Rivers, Estuaries and Coastal Waters Using Innovative Artificial Mussel (AM) Technology- Bangladesh Model

Md Maruf Hossain, Golam Kibria, Dayanthi Nugegoda, TC Lau and Rudolf Wu January 2015

2


.

2. Authors Profile

Professor Md .M. Maruf Hossain, University of Chittagong, Bangladesh Education: Ph.D (University Science Malaysia, Penang , Malaysia) Research interests: Marine & coastal pollution, ecotoxicology (persistent & lipophilic pollutants and it biomagnification in marine food chain), environmental impact assessment (EIA) in different development activities, benthic pollution, water quality assessments, marine biodiversity conservation, biosecurity in coastal aquaculture. Webinks: https://www.researchgate.net/profile/DrMdMMaruf_Hossain http://imsfcu.ac.bd/mmmh/ Email: [email protected]

Dr Golam Kibria, RMIT University, Australia Education: Ph.D (Victoria University of Technology, Melbourne, Australia) Research interests: Climate change, pollution, wetlands, environmental water, ecotoxicology, dams/reservoirs, water quality, ecological risks assessment, arsenic, pesticides, herbicides, trace metals, EDCs, dioxins, pharmaceuticals, microbial pathogens, biotoxins Webinks: https://www.researchgate.net/profile/Golam_Kibria7 http://www.sydneybashi-bangla.com/Articles/Golam_Kibria_main_page.htm Email: [email protected]

Professor Dayanthi Nugegoda, RMIT University, Australia Education: Ph.D (University of London, UK). Research interests: Ecotoxicology, endocrine disrupting chemicals, cyanobacterial toxins; enzymes and hormones, dryland salinity, metals & salinity, nutrient pollution, aquaculture, water quality Webinks: https://www.researchgate.net/profile/Dayanthi_Nugegoda http://www.rmit.edu.au/staff/dayanthi-nugegoda Email: [email protected]

Chair Professor TC Lau, City University of Hong Kong Education: Ph D (University of Hong Kong, Hong Kong) Research interests: Inorganic synthesis, kinetics & mechanisms of inorganic redox reactions, metal catalysed oxygenation & nitrogenation of hydrocarbons, synthesis & magnetic properties of coordination polymers. Weblinks: http://www6.cityu.edu.hk/bhdbapp/deptweb/profile/ptcl.html Email: [email protected]

Chair Professor & Director Rudolf Wu, The University of Hong Kong Education: Ph.D (University of British Columbia, Canada) Research interests: Marine environmental research, marine ecology, marine pollution, ecotoxicology, molecular, biochemical, physiological and ecological responses of marine animals to environmental stresses (hypoxia and xenobiotics), mariculture, hypoxia and xenobiotics Weblinks: https://www.researchgate.net/profile/Rudolf_Wu http://www.biosch.hku.hk/staff/rw/rw.html http://scholar.google.com.hk/citations?user=JnVm1AIAAAAJ&hl=zh-TW Email: [email protected]

3


2. Summary The Artificial Mussel (or AM) is an innovative passive sampling technology/device. The technology (AM) is used to monitor or detect or assess risk of micro-pollutants (such as trace/heavy metals including cadmium, chromium, cobalt, copper, iron, lead, nickel, manganese, mercury, uranium, zinc) in freshwater rivers, brackish estuaries and coastal and marine environments. The technology could also be useful in risk assessment for various water utilities including recycled water, treated wastewaters, groundwater bores, irrigation channels and highly polluted water bodies (anoxic or hypoxic). AM is a continuous monitoring tool providing a time integrated accumulation of metals and has proved to be a very reliable tool for monitoring spatial and temporal variation/distribution of pollutants and identifying pollution “hot spots”. As part of ‘Global AM Watch Programme’, the device has been applied in several countries across the globe including Australia, Bangladesh, Brazil, China, Iceland, Philippines, Portugal, Russia, Scotland, South Korea, South Africa, and the USA. This manual entitled ‘A Training Manual for Assessing Pollution (trace/heavy metals) in Rivers, Estuaries and Coastal waters-Using Innovative “Artificial Mussel (AM) Technology” - Bangladesh Model is the outcome of field and practical experiences gained by the authors while assessing risk posed by trace/heavy metals to water quality, biodiversity and human heath in Bangladesh. The manual has been prepared to assist scientists, environmental regulators, NGOs and general communities to handle, deploy and retrieve “artificial mussel” passive samplers in freshwater rivers, brackish water estuaries and coastal waters for monitoring or risk assessment. It provides simple but step-wise procedures of deployment, retrieval and handling of AM. Images/photos are included for easy use of the training manual (images included in the appropriate section). The manual also includes procedures for storage of AM, occupational, health and safety (OHS) requirements during deployment and retrieval of AM, shipment of AM, other water quality parameters and information requires to be collected during AM watch program.

4


Contents 1.Authors profile

4

2.Summary

5

3. Introduction

7

4. What is an Artificial Mussel or AM?

8

5. Merits of using AM passive sampling method for metals monitoring

9

6. Selection of monitoring sites for AM deployment

11

7. Duration of monitoring in AM watch programme

12

8. Storage of AM

12

9. Occupational health and safety requirements (OHS) in AM watch programme

13

10. Reconnaissance survey, safeguard and set up of AM monitoring/watch site

13

11. Deployment of AM

14

12. Retrieval of AM

16

13. Water quality monitoring and other information to be collected

17

14. Sampling frequency in AM watch programme

18

15. Use of native biota (LM) in AM watch programme

18

16. Significance of three ways of monitoring-AM, LM and SW

19

17. Shipment of AM/LM

19

18. Analysis

20

19. References

20

20. Glossary

22

Artificial mussel monitoring program - the way to go both Australia and Bangladesh

24

5


3. Introduction The Institute of Marine Sciences and Fisheries, University of Chittagong, Bangladesh has been using the “Artificial Mussels” (AM) technology since 2013 to assess the risk posed by trace metals/heavy metals on water quality, aquatic biodiversity, and human health in rivers, estuaries and coastal waters of Bangladesh. The AM technology has been used to monitor the spatial and temporal variation of metals in natural waterways and identifying metal pollution “hot spots” in Australia (Kibria et al. 2010; Kibria et al. 2012a), Hong Kong (Wu et al. 2007), Iceland (Leung et al. 2008), Portugal (Gonzalez et al. 2011), Scotland (Leung et al. 2008), South Africa (Degger et al. 2011) and currently being used in South Korea and Russia. As part of ‘Global AM Watch programme’ and international research agreement (research collaboration between the scientists of the University of Chittagong, Bangladesh (IMSF); RMIT University, Australia; the City University of Hong Kong and the University of Hong Kong), IMSF received AM technology and training and support (as part of the research agreement) and has developed a procedure for deployment and retrieval of AM in waterways of Bangladesh. This procedure includes:       

Handling and storage of AM Setup of monitoring sites for AM Preparation of bags for AM Deployment of bags Retrieval process of AM Shipment of AM Occupational, health and safety requirements (OHS) during field monitoring with AM

This manual highlights the operations IMSF, Chittagong University undertakes during deployment and retrieval of AM in rivers, estuaries and coastal waters of Bangladesh.

6


4. What is an Artificial Mussel or AM? The “Artificial Mussel (AM)” passive sampler is a device that collects or accumulates metals through a diffusion barrier onto a sorbent medium. The device (AM) consists of nonpermeable Perspex tubing (60 mm x 25 cm) in which 200 mg Chelex-100® resin (50-100 mesh from Bio-Rad) is suspended in 8 mL seawater/freshwater inside the tubing (see Figure 1). Both ends of the tubing are further capped by a layer of polyacrylamide gel (thickness: 1 cm), to protect the gel from possible mechanical damages (Figures 1 and 2). Water diffuses through the polyacrylamide gel into the chelax-100 (metal binding agent) from which the complexed metals can later be extracted (see Figure 1). After several weeks, the chelating agent is sampled to determine its metal content.

Figure 1: A schematic diagram showing the design of Artificial Mussel chemical structure of chelex-100 is shown in the inset (Wu et al. 2007).

Figure 2: ‘Artificial mussels‟ device used in metals monitoring, Bangladesh (Kibria et al. 2012a)

7


5. Merits of Using AM Technology Traditional monitoring of metals in the aquatic environment involves determining and comparing metals in water, sediment and biota, but each method has its own problems and limitations (see Table 1). For example, temporal variations in metal concentrations in water are typically large, which often require frequent sampling and analysis that are not cost effective. Bio-monitoring has been used extensively to monitor metals in the last two decades, the notable example of which is the ‘Global mussel watch program’ (Goldberg et.al. 1978; Hossain 1994; Tanabe et.al., 1997; Hossain et.al, 1999, 2000, 2002; Hossain 2004; Kimbrough et al. 2008). The AMs can be deployed in a range of situations (e.g. toxic, anoxic waters) where biomonitoring organisms are not available. Moreover, AM is a reliable tool for monitoring metals in waterways and could be useful in risk assessment for various water utilities including recycled water, treated wastewaters, groundwater bores, rivers, and irrigation channels (Kibria et al. 2010). Until recently there were no reliable time integrated techniques to assess metal concentrations in water which could be used to assess the risk concentrations with respect to water quality guidelines. However, recently Wu et al., (2007) developed an ”Artificial Mussel‟ (AM) technology which have been used to monitor different toxic metals (e.g. Cd, Cr, Cu, Hg, Pb, Zn) in different water body of Australia, Europe, South Africa and China (Leung et al. 2008; Degger et al. 2011; Gonzalez-Rey et al. 2011; Kibria et al. 2012a). This new device is a cost effective monitoring tool which provides a time-integrated concentration of metals in the aquatic environment during the deployment period. Both laboratory and field studies showed that this newly developed chemical device confers significant advantages compared with traditional monitoring techniques using mussels, water and sediment (Wu et al. 2007; Leung et al. 2008; Degger et al. 2011; Gonzalez-Rey et al. 2011; Kibria et al. 2012a).

8


Table 1: A comparison of sampling/monitoring of trace/heavy metals by different techniques (Kibria et al. 2012b). Methods of sampling/monitoring Spot water sampling/monitoring

Bio-monitoring (using live mussels) (e.g. Perna veridis, Mytillus edulis etc.)

Artificial mussels (AM) technology

Pros

Cons

- Quick - Analysis straight forward.

- Snap shot. - Requires frequent sampling/monitoring. - Time consuming. - Costly. - Routine monitoring typically determines total metals but not-bio-available or toxic fractions. -Do not provide spatial and temporal variation in pollutant concentration (e.g. metals)

- Uptake both bio-available and toxic fractions. - Indicate both spatial and temporal variations.

- Continuous monitoring. - No power or energy required. - Metal accumulation/uptake is not affected by biotic and abiotic factors. - Less Affected by salinity & temperature - A standard tool for all waters worldwide (fresh, sea, estuarine, recycled/waste water). - Indicate both spatial and temporal variations. - Can be deployed where bioindicator organisms are not available. - Simple to handle, deploy and retrieve - Simple to analyse - Uptake both bio-available and toxic fractions. - Comparisons valid between sites without calibration if water has similar properties. - identify micro-pollutants “hot spots”.

9

- Requires killing of animals. - Metals accumulation affected by abiotic and biotic factors. - No standard mussel’s species are available for worldwide use. - Some pollutants can be eliminated by the test animals. - Requires translocation of mussels from lab. to field or local mussel species. - Requires complex analysis of biological samples. - Extensive calibration studies are necessary to characterize the uptake of chemicals into a passive sample.


6. Selection of Monitoring Sites for AM Deployment Before deployment of AM in rivers, estuaries and coastal water, the following aspects should be considered as highlighted below: 6.1: The objectives of the investigation should be clearly identified and robust. 6.2: A reconnaissance survey should be conducted to identify the potential high, medium and low risk/impacted sites (e.g. most impacted sites could be sites close by industrial, domestic and agricultural wastes discharge points). 6.3: A survey should be conducted to identify reference/control/least impacted sites (e.g. least impacted or pristine sites could be (a) an upstream of a river without agriculture farming, or domestic or industrial waste discharge points, mining or human habitation; or (b) a natural forest area without human habitation, recreational and agricultural activities). 6.4: If the objectives are to identify “hot spots”, a mix of potential high, medium and low risk sites plus at least one reference/control/least impacted sites should be included in the AM watch programme. 6.5: If the objectives are to protect the ‘most ecologically sensitive sites’ of high conservation values, such as RAMSAR sites or Ecologically Critical Areas ( ECA’s) & Protected water bodies or Marine Protected Areas (MPAs) or ‘Sanctuaries/Reserves’ as declared by Govt. of Bangladesh such sites should be the prime sites for such monitoring. (http://www.mincos.gov.au/__data/assets/pdf_file/0019/316126/wqg-ch3.pdf) 6.6: If the objectives are to protect the aquatic biodiversity (e.g. native fish/biota) or the environment, slightly to moderately disturbed systems and highly disturbed systems should be included in AM watch program (see http://www.mincos.gov.au/__data/assets/pdf_file/0019/316126/wqg-ch3.pdf). Further information regarding aquatic ecosystems monitoring and assessment protocol can be found in ANZECC and ARMCANZ (2000), chapter 7; http://www.mincos.gov.au/__data/assets/pdf_file/0014/316130/wqg-ch7.pdf).

10


7. Duration of Monitoring in AM Watch Programme The duration of AM monitoring/watch programme can be for a short or long periods (pilot or extended monitoring): 7.1: Pilot Monitoring: If the objectives are to collect some preliminary or baseline data and information, a pilot study can be conducted for at least for 3 months by selecting few possible risk sites (possible most impacted, medium impacted, less impacted and least impacted). This pilot or preliminary study will help to trial the AM technology and provide information as to whether trace/heavy metals are a potential problem in an area and whether any further or long term investigations is required etc. 7.2: Extended Monitoring: If the objectives are to assess the effects of pollutants (metals) in an aquatic environment or ecologically sensitive sites (Ramsar sites in Bangladesh such as Tanguar haor, Sunamganj; or Sundarbans, Khulna) or St Martins Coral island, Bay of Bengal, Bangladesh or aquatic biodiversity (e.g. native fish, other native aquatic organisms), or human health - a long term AM watch programme should be run for at least for one to two consecutive years or more. Such study will also generate data on climate variability (dry vs. wet years /or between six seasons of Bangladesh such as summer (14 April-14 June), monsoon/rainy (15 June-17 August), Autumn (18 August-18 October), Late Autumn (19 October-16 December), Winter (17 Decemebr-13 February) and Spring (14 February-13 April) impacts on inputs, transport and bioavailability of trace (toxic metals) including spatial and temporal variations.

8. Storage of AM AMs should be stored in MilliQ (deionized) water or distilled water until deployment in the field in ambient temperature (see Figure 3).

Figure 3: AMs are stored in MilliQ (deionised) water before deployment in the field.

11


9. Occupational Health & Safety (OHS) Requirements It is essential to conduct a site specific risk assessment and take appropriate OHS measures during deployment and retrieval of AM. Accordingly, a site specific risk assessment be conducted and safe work instruction (risk control measures) is to be developed for all the AM watch/sampling sites. This may include working several people (4-5) with life jackets during deployment and retrieval of AM (as experienced during deployment and retrieval of AMs in estuarine and coastal monitoring sites of Bangladesh because of strong tidal action/current and rough sea). Furthermore mechanized good quality boat/ fibre glass boat, carrying a first aid kit, a snake bite kit, sunscreen, insect repellent and personal protective clothing as and when required (sun hat, high visibility vest, swimming life jacket, waders, PVC gloves, lab gloves, safety glass, gum boots, long sleeved shirt and trousers, sturdy footwear, mobile phone) would be essential. Prior weather forecast information/radio weather news (whether the sea is rough or clam), and tide information of the estuarine and coastal sites being monitored are very important.

10. Reconnaissance Survey, Safeguard and Setup of AM Monitoring/Watch Site 10.1: Safeguard of the Site: It is essential that a preliminary survey be conducted to select and identify the risk sites/monitoring sites (see section 6). Once we selected a site in Bangladesh, we had several meetings with the local communities and stakeholders (including govt. authority and local elected representative/s) to inform them about the objectives of monitoring/research, their cooperation in accessing the selected sites during sampling and to be vigilant of our monitoring sites. In addition, we have also hired (paid) local man to take care of AM containing deployed baskets (The duration of each deployment of AM was for 4 weeks). 10.2: Equipment Required: Boat (access to a boat), buoy (access to a buoy), bamboo poles (could be 12-16 feet or 366 cm to 488 cm; depending tidal heights and current etc.), cable ties, deionised water/distilled water, cotton wool/ sponge, metal wire, measuring tapes, name tags, nets, permanent markers, plastic bottles, plastic boxes/baskets with mesh, pen & pencil, pliers, red flags, resealable bags, rubber bands, small rope (cords-polypropylene braded), rocks, red flags, rope (thick), rope (thin), rope (cords-polypropylene braded), strong structures (such as culvert to attach AM baskets), and side cutter. 10.3: Procedure: Baskets containing AMs to be deployed should be attached/tied into a buoy or culvert or bamboo poles (digging strongly into the mud of water (in the bottom) of river, estuary or coastal water in such a distance from the land mark, not to be affected by tide and made sure that AM always remain at least 1 meter below the water surface (see section 11 for deployment of AM).

12


11. Deployment of AM Field deployment of AM procedures and protocols include placing of AM (at least three (03) replicate AM per site) in a plastic basket (17.8 x 16.8 x 15.6 cm) or cage or nylon mesh bag with mesh opening of 5-10 mm minimal for exchange of water (Figure 4a). A piece of rock should be attached under the basket to facilitate sinking of the basket containing AMs (Figures 4b). Strong nylon rope should be passed through the top corners of the AM containing box (Figures 4c and 4d). After that, AM box should be deployed under 1m depth of water (1m below the lowest low tide line level) by attaching the basket either onto a structure of a bridge or culvert or buoy or jetty pillars or bamboo poles (depending on the site locations and availability of the structure at the selected sites) (Figures 4e and 4f).

11.1: Deployment of AM (steps)

4a.Replicate AMs were placed in the basket tied with cable ties

4b. Weight/rock attached at the bottom of the basket to facilitae sinking at least 1 meter below the surface of the water column.

4c. Nylon rope passed through the top corners of the AM basket

4d. Nylon rope passed through the top corners of the AM basket

4e: Bridge structure can be used to attach AM containing baskets

4f: Bamboo pole can be used to attch AM containing baskets (see also 5a to 5c)

Figure 4: Steps in deployment of AM

13


11.2: Deployment of AM in Creek/Canal/River/Sea

Figure 5a: AM can be deployed in a shallow and narrow creek/canal/river/drain by fixing wooden/bamboo poles on both side of the creek/river/drain and passing a rope in between the poles and hanging down the basket at a desired point and operating from the shore.

Figure 5b: AM can be deployed in a tidal river/sea by fixing strong bamboo poles/wooden logs at a desired location or attaching to a behind net. Care should be taken that deployed AM basket is always 1m below the depth of low tide level.

Figure 5c: AM can be deployed by using floating buoys

14


12. Retrieval of AM AM retrieved from the field needs to be properly labelled for easy identification, storage and shipment, which requires the following materials such as cotton wool/sponge, resealable bags, name tags (showing the source, date, water type etc.) and rubber bands.

12: Retrieval Process Each batch of AM deployed in rivers, estuaries and coastal waters should be retrieved at the end of a four weeks (28 day) interval. It is important to retrieve, label and store AMs from each site separately (DO NOT retrieve in bulk and then attempt to accurately label each AM). Baskets containing AM can be covered with silt and algae that should be rinsed with the site water. After rinsing, each AM is wrapped within a wet sponge, with identification tags included inside each whirl pack bag before shipment to a laboratory. The following procedures should are followed: a. The surface of the AM should be rinsed with the site water to remove attached fouling organisms such as algae or other debris (if any) (Figures 6a and 6b). b. Occupational Health and Safety (OHS) equipment should be used when handling AM (globes, apron, masks) to reduce cross contamination of AM samples and for safety of researcher (Figure 6d) c. Sponge/cotton pad soaked with water from the site and wrapped separately around each individual AM, held in place using rubber bands (Figure 6e). d. Each AM is placed in an individual resalable bag (Figure 6f) e. Each bag should be double labelled (with pencilled card inside + water proof label on each resealable bag) (Note: Texta® or pen labels on plastics are easily rubber off and are not recommended). f. The name tag should include date, site name, AM replicate 1, 2, 3 and other information as appropriate etc.

12.1: Retrieval of AM (steps)

6a: Basket containing AM being retrieved

6b: Basket and AM can be covered with silt and aquatic plants (algae) which require rinsing with the site water to remove fouling organisms

6d: It is important to wear OHS equipment when handling AM (globes, apron, masks)to reduce cross contamination of AM samples and for safety of researcher (IMSF Lab., CU).

6c: AM taken out of the basket

6f: AM placed in a resealable bag for shipment to Hong Kong for analysis (City University of Hong Kong)

6e: AM were wrapped in wet cotton/sponges

Figure 6: Steps in retrival of AM

15


13. Water Quality & Other Information to be Collected During deployment and retrieval of AM, some basic water quality parameters of each site should be recorded including surface temperature, water temperature, salinity, dissolved oxygen, electrical conductivity (EC), hardness, pH, rainfall etc. (see Tables 2 and 3). In addition, if the objective is to assess the climate variability impacts on trace/heavy metals inputs, transport and bioavailability, rainfall data in the monitoring site would be required (such as upper catchment), such information can be obtained from local Bureau of Metrological Station. The condition of AM during retrieval (such as whether it is broken or lost) or condition of the basket (e.g. attachment of algae or other aquatic organisms) (e.g. score 1-3, with 1 for minimal algae and 3 to severe) and site information (e.g. access damaged during a recent rainfall or flood or other causes such as vandalism) should be recorded. Table 2: Some examples of site specific average water quality data of 21 sampling sites in Bangladesh. Such data should be collected during retrieval of AM. Sampling sites

Air temp. (ºC)

Water temp. (ºC)

Soil temp (ºC)

Soil pH

Water pH

Salinity (ppt)

Conductivity (ms)

TDS (g/L)

Hardness (mg/L)

Halda (Site 1) Near Madunaghat Bridge Kalurghat (Site 2) Karnafully River Chaktai (Site 3) Karnafully River Sadarghat (Site 4) Karnafully River 15no. Jetty (Site 5) Karnafully River Patenga (Site 6) Mouth of the Karnafully Riiver Khejurtolighat (Site 7) Uttarkattoli coastline Salimpur (Site 8) Fauzdarhat coastline

27.8

28.6

28.1

6.6

7.7

0.4

0.77

0.5

99.16

27.33

28.00

29.2

6.7

7.5

0.3

0.77

0.34

71.25

28.25

28.4

28.7

6.1

6.9

0.74

1.59

1.02

73

28.3

28.7

29.4

6.1

7.0

1.32

3.39

1.56

186.7

24.33

29.4

29.6

6.3

7.22

5.96

10.00

6.7

435

29.3

29.6

29.2

6.25

7.3

6.4

10.7

7.00

615.75

29.3

29.4

29.4

6.3

7.1

11.49

18.33

12.47

701.8

28

29.2

29.2

6.4

7.2

11.71

18.18

12.33

657.5

Bansbaria (Site 9) Sitakundu coast line

32.66

30.4

31.8

6.1

7.2

11.6

17.87

12.68

657.53

Uttar Patenga (Site 10) Patenga coastline Jahajghata (Site 11) Anwara coastline Moheshkhali Channel (Site 12), Cox’s Bazaar Near hatchery zone (Site 13), Sonapara, Cox’s Bazaar Rejukhal (Site 14) , Cox’s Bazaar

34

30.33

32

6.4

7.3

15.88

24.58

17.0

948.71

34.6

30.5

32.23

6.3

7.3

16.6

25.72

17.95

1053.2

29

28

30

6.4

7.4

27

43.38

29.27

1656.48

29.5

28

30

6.2

7.1

28

44.21

30.23

1765.43

28.5

27

29

6.3

7.2

23

35.23

24.82

1374.01

Bastuhara boropol (Site 15), Buriganga River Near Babubazarbridge (Site 16), Buriganga River

31.6

27.83

27.5

5.2

6.9

0.56

1.10

0.75

176

33.16

31.5

30

5.7

6.8

0.61

1.15

0.78

160.11

Kamringir Char (Site 17), Buriganga River Khulna (sites 18-21)

33.33

29.33

32

5.66

6.8

0.63

1.25

0.84

186.83

Mirerdanga Kheyaghat (Site 18), Bhairav River

35.7

31.25

33.5

6.22

7.5

6.75

9.76

8.78

459.89

Hardboard mill Kheyaghat (Site 19), Bhairav River Mathavanga (Site 20) Rupsa River Near Mongla port (Site 21), Passur River

36.5

31.3

34.37

6.27

7.43

7.27

11.68

8.29

514.63

34.7

33

37.67

6.27

7.5

8.32

13.42

9.43

581.44

37.25

33

37.67

6.4

7.2

10.22

16.8

11.36

685.55

Chittagong (sites 1-14)

Dhaka (sites 15-17)

16


Table 3: Average rainfall in Chittagong during 2013 (www.weather-and-climate.com) Average precipitation in Chittagong (mm)

Jan 6

Feb 28

Mar 63

Apr 151

May

Jun

265

533

Jul 598

Aug 519

Sep

Oct

Nov

180

55

321

Dec 16

14. Sampling Frequency in AM Watch Programme Three to six replicate AM should be placed at each site in a basket or cage or nylon mesh bag and be retrieved after four weeks interval.

15. Use of Native Biota in AM Watch Programme In addition to deployment of AM passive samplers at a site, a similar number of live native mussels (LM) can also be stocked (matching AM numbers; an example 3 to 6) in separate baskets or cages or nylon bags at the same site, where AM were deployed (see Figure 7). This is called bio-monitoring or active monitoring and such strategy may or may not be feasible or possible because of the non-availability of native mussels or poor survival of live mussels in baskets/cages. In addition, live mussels require depuration of toxic metals prior to deployment (at least 1-2 weeks) which can be done by keeping them in tanks for few days. LM should be selected based on biological characteristic such as abundance, hardy in nature (e.g. tolerance to low dissolved oxygen). Inclusion of biota will help to assess the bioaccumulation potential of metals to native biota, threats (such as metals) it poses to native biota and risks to human consumers (public health risk) via eating metals contaminated seafood. It will also allow comparing the time integrated accumulation of heavy metals in artificial mussel (passive samplers) with that in native mussels (biota) (i.e. accumulation of metals in biota, i.e. LM vs. AM device). It is essential that approval is received for translocations of live aquatic organisms before such live mussels stocking can be done (from appropriate authorities since there could be some restriction in translocation native species into new areas). It is further suggested that native mussels to be stocked should be originated from the same river or estuaries or coastal areas to avoid any genetic variability between populations and transfer of any diseases etc. into a new location.

7.1: Live mussels (LM) Perna viridis collected from Cox’s Bazar, Bangladesh for experiments/deployment with artificial mussels (AM).

7.2: LM collected from Coxes Bazar, Bangladesh for experiments/deployment with AM.

7.4: LM and AM deployed in Coastal areas of Bangladesh (Mosheskhali, Cox’s Bazar; October 2014)

7.3: LM being sorted out for experiments/deployment with AM

Figure 7: Locally available live mussels, Perna viridis (collected from Moheskhali, Cox's Bazar, Bangladesh) were deployed at selected coastal areas of Bangladesh with artificial mussels (AM).

17


16. Significance of Three Ways of Monitoring- AM, LM and SW (a) Use of novel AM technology: Will provide the most accurate spatial and temporal estimations of pollutants. AM will also provide information relating to bio-available and toxic fractions of pollutants. AM will enable identification of pollutants “hot spots” (Kibria et al. 2012b). (b) Use of live native (LM) mussel: Will provide both spatial and temporal variations of pollutants in aquatic biota but not as accurate as AM (since accumulation of pollutants in biota can be significantly affected by chemical and biological factors). However, bioaccumulation in LM can provide an indication of threats posed to local biota as well as to human consumers (from eating metal contaminated seafood). LM is also an additional check of results obtained with AM (Kibria et al. 2012b). (c) Spot water (SW) samples: It is a validation and quality control exercise. Data of spot water will be an additional check-up of data obtained with AM and LM of a site (i.e. whether it matches or differs). Spot water sampling data will also enable comparisons with the recommended aquatic ecosystems guideline values for metals. There will be a comparison amongst spot, LM and AM in determining risk. However, spot water sampling is a snap shot of pollutants, determines total metals but not-bio-available or toxic fractions and do not provide spatial and temporal variation in metals (Kibria et al. 2012b).

17. Shipment of AM After retrieval, each AM is wrapped with a wet sponge with identification tags and custom declaration form included inside each whirl pack bag before shipment to Hong Kong for analysis. AM shipped to Hong Kong via Fedex express courier (Figure 8).

Figure 8: AM finally packed in Cartoon and ready for shipment to Hong Kong for analysis via courier service (FedEx).

18


18. Analysis Analysis of environmental samples should be done in any ISO 17025 accredited laboratory for the analyte in question. The laboratory should be equipped with either flame atomic absorption spectrophotometry (FAAS) and/or inductively coupled plasma atomic emission spectrometry (ICP-AES).

18.1: Analysis of Artificial Mussels (AM) The contents of each individual AM (chelex resins) will be emptied into a sintered glass filter followed by eluting two times with 12.5 mL 6 M HNO3 (analytical grade). The elutriant will be made up to a known volume with deionized double distilled water and the concentrations of Cd, Cr, Cu, Pb and Zn will be determined by flame atomic absorption spectrophotometry (FAAS; Shimadzu 6501S) and inductively- coupled plasma atomic emission spectrometry (ICP-AES; Perkin- Elmer Plasma 1000). Concentrations of metals in AM will be expressed as μg/g of chelex (Wu et al. 2007; Degger et al. 2011; Kibria et al. 2012a).

18.2: Analysis of Live Mussels (LM) Live mussels will be dissected with a knife, and their byssal threads will be removed. The soft tissue will be rinsed with Milli-Q-water, weighed and subsequently dried in an oven at 60oC to a constant weight. The dry weight will then be determined before acid digestion in a block digester, using 30% hydrogen peroxide and 70% nitric acid (1:1 v/v). Metal concentrations in the soft tissue of the digested samples will be determined using FAAS or ICP-AES as appropriate. Concentrations of metals in mussel tissues will be expressed as μg/g dry tissue (Wu et al .2007; Degger et al. 2011).

19. References Degger, N., V. Wepener, B.J.Richardson and R.S. Wu (2011). Application of artificial mussels (AMs) under South African marine conditions: a validation study. Mar Pollut Bull. 63(5-12):108-118. Goldberg, E.D., V.T. Bowen, J.W. Farrington, G. Harvey, G.H. Martin, P.L. Parker and R.W. Riserbrough (1976). The ‘Mussel watch. Environ. Consrn. 5: 101-125. Gonzalez-Rey, M., T.C. Lau, T. Gomes, V.L. Maria, M.J. Bebianno and R. Wu (2011). Comparison of meta Accumulation between 'Artificial Mussel' and natural mussels (Mytilus galloprovincialis) in marine environments. Mar Pollut Bull. 63(5-12):149-53. Hossain, M.M (2004). An assessment of the ability of natural stocks of Green mussels (P.viridis), clam (Meretrix meretrix) & oyster (Crassostrea sp.) to meet international standards of food safety regarding unnatural contaminants. Presented in technical session (post-harvest technology sect.) of the Nat. Seminar on the ‘Fisheries Education & Research Fair’, 04, in BIAM Auditorium, Dhaka, Bangladesh, 19-20 July, 2004, organized by Bangladesh Fisheries Research Forum (BFRF) & supported by DFID (SUFER Project), UK & UGC, Bangladesh (Key findings of the SUFER Funded research project).

19


Hossain, M. M. Z.B. Din and S. Ibrahim (2002). Spatial distribution of organochlorine pesticides in three bivalve species collected at the west coast of Malaysia. Chittagong University Studies (Sci.). 27(2): 4146. Hossain, M. M, Z. B. Din and M. S. Ibrahim (2000). Lipid-magnification of persistent organochlorine pesticides in 3 bivalve species collected along the west coast of peninsular Malaysia. Chittagong University Studies (Sci.). 24 (2): 111-119. Hossain, M. M. Z. B. Din & M. S. Ibrahim (1999). Bio-concentration and Availability of Organochlorine Pesticides (OCP) in Bivalves and Marine Sediment from the West Coast of Peninsular Malaysia. Presented in ‘International Conference on the International Oceanographic Data & Information Exchange in the Western Pacific (IODE-WESTPAC)’, 1999, held in Langkawi, Malaysia from 1-4th Nov 1999. Published in Proc. of the Conf., Japan Oceanographic Data Center (JODC), Japan. p. 323331 Hossain, M. M (1994). Concentration of total Mercury in marine shrimps and fish species from the Bay of Bengal, Bangladesh. “Marine Research” J. of Marine Biology, Pakistan. Vol. 3 (2): 27-32. Kibria G., T.C. Lau and R. Wu R (2012a). Innovative ‘Artificial Mussels’ technology for assessing spatial and temporal distribution of metals in Goulburn-Murray catchments waterways, Victoria, Australia: Effects of climate variability (dry vs. wet years). Environment International journal.50: 38-46. Kibria, G., G. Rose, T. C. Lau and R. Wu, R 2012b. A Training Manual for Deployment, Retrieval and Handling of Innovative Artificial Mussel Passive Samplers for Trace Metals Monitoring in Rivers, Creeks and Channels- Australian Model. The Training Manual has been prepared under a research collaboration agreement between Goulburn Murray Rural Water Corporation, Tatura, Australia, the City University of Hong Kong, and the University of Hong Kong. G-MW docs #3316583; 3 tables, 13 figures, 38p. https://www.researchgate.net/publication/261178237_A_Training_Manual_for_Deployment_Retrieval_and_Handling_of_Innovative_Artificial_Mussel_Passive_Samplers_for_Trace_Me tals_Monitoring_in_Rivers_Creeks_and_Channels-_Australian_Model

Kibria, G., G. Allinson, V. Pettigrove, P. Slessar, T.C. Lau and R. Wu (2010). Monitoring trace metals in Nort and Central Victorian Waterways, Australia, using Artificial Mussel (AM) Technology (2009-2010). Report prepared under a research collaboration agreement between Goulburn Murray Rural Water Corporation, Tatura, Australia, the City University of Hong Kong, the University of Hong Kong, and the Department of Primary Industries, Werribee, Victoria, Australia. 35p. https://www.researchgate.net/publication/273260081_Monitoring_trace_metals_in_North_and_Central_Victorian_Waterways_Victoria_Australia_%28Research_collaborat ion_between_GM_Water_Australia_the_University_of_Hong_Kong_and_Department_of_Primary_Industries_Victoria_Australia%29

Kimbrough, K. L., W.E. Johnson, G.G. Lauenstein, J.D. Christensen and D.A. Apeti (2008). An assessment of Two decades of contaminant monitoring in the Nation's Coastal Zone. Silver Spring, MD. NOAA Technical Memorandum NOS NCCOS, 74. 105 pp. Leung, K. M., R.W. Furness, J. Svavarsson, T.C. Lau TC and R.S. Wu RS (2008). Field validation, in Scotland and Iceland of the artificial mussel for monitoring trace metals in temperate seas. Mar Poll. Bull. 57 (612):790-800. Phillips, D.J.H (1985). Organocholorine and trace metals in Green lipped mussels (P.viridis) from Hong Kong waters: test indicator ability. Mar. Ecol. Prog. Ser. 21: 251-258. Tanabe, S., S. Kan-atinelap, J. Sanguansin, S.M. Tubucanon, S.M. and Hungspreugs, M (1997).Contamination Of butylin compounds and organochlorine residues in Green Mussels (P. viridis) from Thailand coastal waters. Env. Poll. 97: 79-89. Wu, R.S. S., T.C. Lau, W.K. M. Fung, P.H. Koa and K.M.Y. Leung (2007). An artificial mussel for monitoring heavy metals in a marine environments. Environmental Pollution, 145, 104-110.

20


20. Glossary Artificial Mussel: or AM (see passive sampling). Bio-available: the degree and rate at which a substance (chemical, toxicant) is absorbed into a living organism or is made available at the site of physiological activity. Bio-monitors: a species that is sensitive to, and shows measurable responses to, changes in the environment, such as changes in pollution levels. Biotic: relating to living organisms. Calibration: to check, adjust, or determine by comparison with a standard. Chelex-100 resin®: is a chelating material which has the ability to bind transition metal ions. Chelating agent: certain organic compounds which are capable of forming coordinate bonds (with metals through two or more atoms of the organic compound); such organic compounds are called chelating agents. The compound formed by a chelating agent and a metal is called a chelate. Ethylenediaminetetraacetate, (-O2CH2)2NCH2CH2N(CH2CO2-)2, (EDTA) is a common chelating agent. CityU: City University of Hong Kong. Concentration: The strength of a solution; number of molecules or mass (e.g. in μg/l) of a substance in a given volume. Deployment: placing of AM. Electrical conductivity or EC: Electrical conductivity (EC) estimates the amount of total dissolved salts (TDS), or the total amount of dissolved ions in the water. Heavy metals : Are a group of metallic elements with an atomic number greater than 20 e.g. mercury, chromium, cadmium, and lead characterized by the ability to form co-ordination bonds with organic chelates and anions, and can damage living things at low concentrations and tend to accumulate in the food chain. HKU: the University of Hong Kong. Limit of detection: is defined as the concentration at which an analyte can be identified but not quantified. Limit of reporting (LOR): is defined as the concentration at which an analyte can be identified and quantified with 99% certainty. Milli-Q or Type 1 water: also called high purity water, refers to water that has been purified and deionized to a high degree by water purification systems. Micro-pollutants: are compounds which are detected in the concentration range of ng/L up to μg/L in the environment. Passive sampling: A device that collects or accumulates pollutants (e.g. Heavy metals) by diffusion onto a receiving phase usually separated from water by a membrane. Polyacrylamide: Polyacrylamide (poly(2-propenamide) is a polymer (-CH2CHCONH2-) formed from acrylamide subunits that can also be readily cross-linked. RAMSAR sites: Wetlands that is important for conserving biological diversity. The Convention on Wetlands of International Importance (the RAMSAR Convention) was signed in Ramsar, Iran on 2 February 1971. The

21


Ramsar Convention aims to halt the worldwide loss of wetlands and to conserve, through wise use and management, those that remain. Retrieval: removal of AM from the rivers Spatial: of or relating to space Temporal: of, relating to, or limited by time Toxic: of, pertaining to, affected with, or caused by a toxin or poison Trace metals: Metals with a concentration in water between μg/L to mg/L.; e.g. calcium and magnesium associated with essential elements Translocation: Translocation is the movement of living organisms from one area with free release in another. This includes intentional and unintentional

22


Artificial Mussel Monitoring Programme - the way to go both Australia and The Artificial Mussel program (AM) is an innovative passive sampling (i.e. continuous monitoring) technique for monitoring program of micropollutants (concentrations range from nanogram to microgram per litre) such as trace/heavy metals in waterways (e.g. rivers, creeks, estuaries, coastal waters, oceans). The initiative is part of a Global AM Watch program being run in eight countries, including Australia, Bangladesh. The technology can be used to assess and monitoring risk posed from trace/heavy metals including cadmium, chromium, cobalt, copper, iron, lead, nickel, manganese, mercury, uranium, zinc. Results from extensive studies carried out in Australia can be accessed via the following weblinks: https://www.researchgate.net/profile/Golam_Kibria7

Dr Golam Kibria, demonstrating AM technology to graduates of the Commonwealth Department of Environment, Water, Heritage and the Arts (DEWHA), Australia during their field visit to the Kangaroo Lakes, Victoria, Australia (25 August 2010).

The Australian study found elevated or higher levels of metals (copper, zinc) in waterways close to intensive orchards (pome and stone), vineyards and areas growing olives and tomatoes. The programme has received both national and international interest. GM Water, Australia provided AMs to Melbourne Water and North Central Catchment Management Authority (NCCMA), Victoria, Australia to monitor trace metals in their waterways as part of research collaboration with GM Water. The deployment and retrieval techniques for AM developed by Golam Kibria (to suit GM Water catchment) used in training of African delegates in Monaco in August 2010 to monitor water quality in African catchments (sponsored by the International Atomic Energy Agency). GM Water's use of the AM technology was recognised by Commonwealth Scientific, and Industrial Research Organisations (CSIRO), Australia and the Australian Society for Ecotoxicology (ASE) at a recent conference (13th ASE, Adelaide, 2009). Dr Golam Kibria received an award at an ASE conference related to a research work completed on AM monitoring in Australia. GM Water also demonstrated the benefits of AM technology to graduates of the Commonwealth Department of Environment, Water, Heritage and the Arts (DEWHA) during their field visit to the Kangaroo Lakes (25 August 2010; see the photos above). DEWHA graduates were impressed by the innovative technology used by GM Water to assess the risks of micropollutants in waterways. The Australian study found that AM is a reliable tool for monitoring metals in waterways and could be useful in risk assessment for various water utilities including recycled water, treated wastewaters, groundwater bores, rivers and irrigation channels. In Bangladesh, the Artificial Mussels Technology was used for the first time (2013) (Phase -1) and later in 2014 (phase-2) to assess the possible threats and risks posed from trace/heavy metals pollution to various sectors including water (water quality, drinking water), ecosystems, biodiversity, fisheries, agriculture, seafood export commodities, livelihoods and human health. As part of global AM Watch programme, a monitoring was conducted in the Halda River, Karnafuli River estuary and adjacent coastal waters of Chittagong, Bangladesh during June 2013-December 2013, and later in addition to coastal area of Chittagong, rivers and coastal areas of Khulna and Mongla (2nd port city), Cox’s Bazar, Mosheskhali and the river Buriganga, Capital city Dhaka, the most polluted river of Bangladesh (Phase-2,an extended program of phase-1, June- Dec.2014). The study generated some new knowledge and information for Bangladesh including the following: (a) successfully deployed and retrieved AM in waterways of Bangladesh; AM deployed in coastal waters in Bangladesh (2013-2014) (b) identified possible metals pollution sources in the studied areas; (c) being retrieved. detected eleven metals (Cd, Cr, Co, Cu, Fe, Hg, Ni, Mn, Pb, Zn, U), in the studied areas, some of which are known to be highly toxic to biota (Cd, Cu, Hg, Pb, Zn, U), are endocrine disruptors (Cd, Pb, Hg, Zn) and carcinogenic to humans (Cd, Cr, Ni, Pb, U); (d) documented the spatial and temporal variation in metal concentrations during the period of investigation (rainy season, autumn and late autumn) and (e) Identified seven metal pollution “hot spots “in the selected rivers, estuary and adjacent coastal waters of Bangladesh. (f) Comparison of uptake mechanisms of metals was also tested for LM (P.viridis) and AM during the investigation period. Further information can be obtained by emailing the following research scientists: Dr Golam Kibria (Leader Global AM watch Programme- Australia, Bangladesh, Russia and South Korea): [email protected] Professor Md Maruf Hossin (Leader Global AM Watch Programme, Bangladesh): [email protected]

23