EnvironmentAsia The international journal published by the Thai Society of Higher Education Institutes on Environment Available online at www.tshe.org/ea/index.html EnvironmentAsia 9(2) (2016) 48-54 DOI Qualitative Assessment and Management of Microplastics in Asian Green Mussels (Perna viridis) Cultured in Bacoor Bay, Cavite, Phillipines Cristian Ryan Argamino and Jose Isagani B. Janairo Biology Department, De La Salle University, 2401 Taft Avenue, Manila 0922 Philippines Abstract Microplastics (> 5 mm) have gained popularity in research and the public eye in recent years. This is due to the fact that they contain persistent organic pollutants (POPs) which pose potential risks to the environment and human health. Bivalves, which are filter feeders, are considered to be good indicators of marine pollution. In this preliminary study, Asian green mussel (Perna viridis), an example of edible bivalve, cultured in Bacoor Bay, Cavite, Philippines was subjected to qualitative analysis to determine the presence of microplastics. Through microscopic analysis, microplastics were found present in the acid-digested mussel soft tissue. A management program is suggested for policy makers and stakeholders to reduce the negative impact of microplastic pollution to both humans and the marine environment. Keywords: microplastics; qualitative assessment; Perna viridis; marine pollution; environmental management 1. Introduction Plastics are popular due to several properties such as weight, strength and cost. The rise of consumption over the past few decades has been a huge challenge in terms of controlling environmental pollution. The increase in the number of plastics amassed in the environment is mainly caused by their inert property (slow degradation rate) and usage (improper disposal of plastic waste) (Nor and Obbard, 2014). Despite their benefits, the contribution of plastic to environmental degradation has been significant in causing damage to marine organisms. Previous studies have shown that plastics (i.e. microplastics) are ingested at different trophic levels which can potentially cause bioaccumulation. This phenomenon can affect biological processes of marine organisms. In addition, food safety and human health are also at risk, if these affected organisms are consumed (Vandermeersch et al., 2015). It has been estimated that up to about 12.7 million tons of plastic still ended up in the ocean in 2010 (Jambeck et al., 2015; Andrady and Neal, 2009; Van Cauwenberghe et al., 2015). The Philippines was ranked No. 3 by mass of mismanaged plastic waste following China and Indonesia. In 2010, plastic marine debris in the Philippines ranged from 0.28-0.75 MMT (Million Metric Tons) per year (Jambeck et al., 2015). Marine plastic debris primarily come from land-based sources (80%) through leakage while only marine aquaculture and fisheries only contribute 20% of these pollutants (Ocean Conservancy, 2015). In recent years, the emergence of a form plastic has been studied for its role in pollution - microplastics. Although there has been no universally accepted definition at the moment (Van Cauwenberghe et al., 2015), individual and group researches have contributed their own description of microplastics. In 2004, microplastic size was defined at around 20 microns (Thompson et al., 2004). In 2009, the size adapted was <5 mm. This characterization of microplastic size was also accepted by the Joint Group of Experts on the Scientific Aspects of Marine Environmental Protection (GESAMP) (Nor and Obbard, 2014). Other researchers have presented microplastics as particles with <1 mm size which is argued to be more instinctive since ‘micro’ refers to the micrometer range (Van Cauwenberghe et al., 2015). As a marine pollutant, microplastics exist in two forms - primary and secondary. Primary microplastics, directly move into water bodies through ground runoff. These are mainly composed of virgin plastic pellets, scrubbers, and microbeads. These plastic forms are abundant in cosmetic products like exfoliants as well as industrial abrasives (Andrady, 2011). On the C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54 other hand, secondary microplastics are degradation products of larger plastics (mesoplastic/macroplastic) which are already in the ocean or seashore. Microplastics in this form are produced via several pathways such as mechanical, photo (oxidative) and/ or biological degradation (Masura et al., 2015). Majority of microplastics in the marine environment are secondary microplastics formed through the weathering of plastic in the seashore. Plastics on beaches have a faster degradation rate due to the higher temperature (of sand) as compared to both plastics deep in the ocean or floating on the water surface (Andrady, 2011). Mussels, locally known as tahong, are generally defined as mollusk bivalves which grow in the wild or through aquaculture. These suspension feeders are considered to be one of the best biological indicators of marine pollution because of various characteristics including their geographical distribution. Their tissues accumulate pollutants due to their feeding mechanism and their tendency to stay attached to surfaces which make them relatively inactive in terms of mobility thus rendering them as good pollution gauges. They can also be easily sampled and thus, allow for frequent experiments and monitoring (Vasanthi et al., 2012; Figueiras et al., 2002; Chase et al., 2001). The species of mussel used in this experiment is Perna viridis (P.vidiridis). P. viridis is a large species of mussel ranging from 8-16 cm (Rajagopal et al., 2006). Their habitat includes hard surfaces (rocks, pilings and floating docks, as well as sandy or muddy bottoms about a foot below the low tide mark) (McGuire and Stevely, 2009). Their diet includes microscopic phytoplankton, zooplankton and suspended organic detritus in the water (Rajagopal et al., 2006). In the Philippines, P. viridis or tahong is considered as one of the seven major aquaculture species (Food and Agricultural Organization, 2006). Aquaculture of tahong can be traced back to 1955 which was initiated by the Philippine Bureau of Fisheries and Aquatic Resources in Binakayan, Cavite (Yap, 1999). Although it is a source of food locally, discretion is advised for consumption due to their tendency for bioaccumulation of toxins substances detrimental to human health (McGuire and Stevely, 2009). Thus, determining the presence of microplastics in these marine species will not only be instrumental to profiling marine pollution but will also serve as a tool in improving guidelines for food safety. In this study, we present our findings regarding the presence of microplastics in cultured bivalves available for human consumption. These particles were positively identified through microscopic analysis and other qualitative criteria. In addition, management procedures according to best practices were also discussed which will aid policy makers in reducing the risk posed by these pollutants to the environment and human health. Figure 1. Map of Bacoor Bay (Google Maps) 49 C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54 Figure 2. Semi-transparent microplastics 2. Materials and Methods 3. Results and Discussion Bacoor Bay-cultured samples (P.viridis) were obtained from Sineguelasan Seafood Terminal in Bacoor, Cavite, Philippines. The mussel samples were harvested on January 2016. Fig. 1 shows a map of Metro Manila and its adjacent provinces. The drop pin marks the location of Bacoor Bay where the samples were cultured and harvested. All glassware were thoroughly cleaned and rinsed with filtered deionized water (Elga PURELAB Flex) to avoid contamination dilution (Van Cauwenberghe and Janssen, 2014). Wet digestion using acid was used to extract the microplastics for the samples (Vandermeersch et al., 2015). Twenty milliliters (20 mL) of 70% HNO3 (UNIVAR) were added to three mussels in an Erlenmeyer flask. Five replicates were prepared. Mussel tissues in acid were left in in the hood for 40 hours to achieve optimum digestion. The samples were then heated until boiling for 15-20 minutes using a hot plate to evaporate the acid and dry the sample. Twenty milliliters of warm deionized water (~80 degrees Celsius) were added to each vessel for dilution (Van Cauwenberghe and Janssen, 2014). Digested samples were then subjected to vacuum filtration using a Buchner funnel and Whatman Filter Paper Grade 1 (11-micron pore size). Filters were dried for 2 hours in an oven at 40 degrees Celsius. Dried filters were analyzed for the presence of microplastics using a Nikon SMZ 745T Stereomicroscope (Van Cauwenberghe and Janssen, 2014; Song et al., 2015). Previous studies confirmed the presence of microplastics from marine samples using different methods. Raman Spectroscopy (Van Cauwenberghe and Janssen, 2014), Fourier Transform Infrared Spectroscopy coupled with either a Microscope or an Attenuated Total Reflectance attachment (Nor and Obbard, 2014), as well as the Fluorescence Microscope (Noren, 2007) were utilized by various studies. The absence of these instruments should not be a hindrance in the analysis of this important class of pollutants. Thus, the following criteria were used in determining the presence of microplastics (Noren, 2007): a) No cellular or organic structures are visible in the plastic particle/fibre. b) Clear and homogeneously colored particles (blue, red, black and yellow) c) If the particle transparent or whitish, it shall be examined with extra care in a microscope under high magnification. Figs. 2 and 3 show examples of microplastics observed in each of the five samples. Microplastics observed in the samples were all found to be < 1mm. Transparent whitish and reddish particles measuring at around 10 to 30 microns (0.1 to 0.3 mm) were observed in Samples 1, 2, and 5. On the other hand, blue fibers (~0.5 mm in length) were observed in Sample 3 and 4. These observations qualify in the abovementioned criteria (Noren, 2007) p c e t y Figure 3. Fiber micropplastics i r e v l 5 t 50 fc (e g ry i mr be hv al b a5 n h a C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54 t Figure 4. Microplastic Managementt i presencef of microplastics m therefore, confirming the M MWith the confirmation w h the presence of in P. viridis. of b 5 microplastics in this preliminary study, ufurther u research on microplastics can now be focused yon)their G s analyticalotools quantitationyand characterization using i n i i such as the Fourier Transform Infrared Spectroscopy u M u n c Microscope and Raman Spectroscopy. Microplastic o m u presence needs to be addressed by policy makers a and a d stakeholders. management program g t is g A suggested a r summarizedain Fig. 4 (GESAMP, 2015). aT r aal The first dimension of the program d e is a d v m Comprehensive Waste Generation Management o o (CWGM) (Fig. 5) which can haveo a big impact h in i decreasing the amount of plastic in oceans. According i n d o to f Jambeck et al. (2015), a 50% decrease in mismanaged n f c Mu waste fromMthe Top 20 ncontributorsp (which includes u the Philippines) will result of u r to 41% fmass decrease e h mismanagede plastic waste by 2025.1CWGM involves l of hotspots a or potential n sourcesl of s the identification pollutant. In this case, microplastics ingested by bivalves can be attributed to sources such as urban inputs, fisheries, and aquaculture. Another aspect of CWGM is life cycle assessment which includes value-chain models and the popular 3R method (reduce, reuse, recycle). Life cycle assessment of plastics particularly the product disposal chain is an important task of management. This assessment will lead to a targeted approach which are prerequisites of an efficient and cost-effective method of reducing microplastic impact (Vegter et al., 2014). The 3Rs in im f m mn particular should be targeted toward reducing marine wd hh dn h o debris by giving economic incentives to stakeholders. 5 s a b s t The last aspecta of this program is the reductions of ) f s eThis can be microplastic input (GESAMP, 2015). G s a of a n implemented through policies i e n r whichteban or regulate primary microplastics. The ny cm M United States y l of America m recently enacted a law “prohibiting the r f manufacture o r m u s and or delivery for introduction d a introduction a ginto g interstate commerce containingt a f r t e off cosmetics f t of rinse intentionally-added plastic microbeads” (Microbeadr a T p o al p o e Free 2015). Microbeads in cosmetic v o WatersmAct, e o l l a products are usually made t upr of polyethylene. Policies o he A of must also drive the reduction and Sprevention S A e n y du o(formed y u secondary microplastics from degradation) n m n i d inc entering the marine environment (Dippo, 2012). d i e a n p u e Research can also e e be driven towards thea reductione of r f rare microplastic input. c r Plastics in a marine cenvironment e hu e c a usually in the1 form of packaging materials. Further e a a n s c c e e either by studies on alternative packaging materials significantly reducing the amount plastic used or redesigning and formulating alternative materials which can be easily and safely degraded to non-threating products (for both human and marine life) will decrease marine litter output. Physical screening and waste treatment technologies are also mitigation options to prevent microplastics from entering not only the marine environment but also groundwater which can pose risks to terrestrial animals and humans (Vegter et al., 2014). Fig. 5. shows Comprehensive Waste Management Program for Microplastics. o t f f r s e e t r e n e a u e Figure 5. Comprehen nsive waste generation g maanagement plan p n l n m i F g l n n t a m i F v s n k 51 t o i p a s k o n m c h n a d o m c h k a e l n o n a e l e C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54 t Figure 6. Risk Percepttion and Manaagement of Miicroplastics a i e Another dimension of the proposed management e a m a m scheme is obtaining feedback from stakeholders on a m m n by microplastic h u their perception of the risks posed n h aand Management) mo contamination a (Risk Perception m h awareness eone the (Fig. 6). It hstarts with promoting e m e impacts of these pollutants to humans and uthe p ostakeholders w and environment o alike. Educating w i t fa t making themi understandfathe hazardsa of microplastic e e contamination tool efor management e is an effective e a e (Dippo, 2012). e Another key aspect ofo communicating l the risks involved is to convert complex terms to a language that can be fully understood by the stakeholders. Developing a management plan for this social, behavioral, and psychological aspect of microplastic management involves surveys, psychological studies, and acknowledging demographic differences. These include regional, cultural, economic, educational, and other variances in behaviour and perceptions (GESAMP, 2015). A comprehensive management framework is shown in Fig. 7. To mitigate the effects of microplastic contamination to marine and human life, policies should be put into place to strengthen the foundation of the measures to be employed. The illustration also shows the pillars of mitigation action and management which include research, implementation, and regulation. Research entities from the government (Department of Science and Technology, Academic Institutions, National Fisheries Research and Development Institute,) are tasked to develop infrastructure, methods, t a n a e m h m a technologies m s prevent microplastic i and that will s i e m u o l entry to the marine environment. Implementation o l u lap of Local tGovernment responsibilityowill fall on the u e Provincial to the Municipal am o Units from the Level a o h m e I headed by the Environmental and Natural Resources p u that policies s I are being e implemented u Officer to ensure s e u h t t m using the developed technologies. Regulatory bodies a tm m m c sucht as the Environmental Management Bureau and m c M e a r h Bureau of Fisheries and Aquatic Resources will act a o r l uh s as anu auditing entity to ensure that effluents and s o marine bodies are checked in terms of microplastic levels. e t m h h m M a o 4. Conclusions Microplastics were found present in Bacoor bay-cultured tahong (P. viridis). This preliminary study confirms a microplastic pollution problem in the Bacoor Bay and Manila Bay area and thus, the need for further research on quantitating and characterizing these pollutants is of utmost importance. Management and mitigation procedures to prevent the entry of microplastics (both primary and secondary) require policy makers, stakeholders, government agencies, non-government organizations, and academic institutions to work in sync to put up a strong and effective management framework with research, implementation, and regulation as its pillars and policy as its foundation. Figure 6. Risk Percepttion and Manaagement of Miicroplastics a i a a m n a h e m m u h o m e e a s m o 52 u m m u l o e a e s i o u o e l t m a h i o I e t m h C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54 t 7. 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