Evaluation of the Interaction Among Microalgae Spirulina sp , Plastics Polyethylene Terephthalate and Polypropylene in Freshwater Environment

The annual plastic production in Indonesia has exceeded 4.6 million tons and accumulated in the aquatic system. Polyethylene terephthalate (PET) and Polypropylene (PP) are the most widely used plastics in manufacture of packaging, fibres, and drinking bottles, etc. The degradation of these plastics to micro sizes leads to environmental threats, especially when the micro plastics interact with fresh water microorganism such as microalgae. Therefore, the study on the interaction between micro plastics and microorganisms is really important. The aim of this study was to evaluate the impact of microplastics on the growth of microalgae Spirulina sp and also to evaluate the contribution of microalgae Spirulina sp to the plastic degradation. The interaction between microalgae and microplastics was evaluated in a 1 L glass bioreactor contained microalgae Spirulina sp and PP and PET microplastics with the size of 1 mm at various concentrations (150 mg/500 mL, 250/500 mL and 275 mg/500 mL) for 112 days. The results showed that the tensile strength of micro plastic PET decreased by 0.9939 MPa/day while PP decreased by 0.1977 MPa/day. The EDX analysis of microplastics showed that the decreasing carbon in PET (48.61%) was higher as compared to PP (36.7%). FTIR analysis of Spirulina sp cells showed that the CO2 evolution of cells imposed by PET microplastic was higher than imposed by PP. The growth rate of Spirulina sp applied by micro plastic was lower than the control and the increase of microplastic concentration significantly reduced the growth rate of algae by 75%. This research concluded that biodegradation has important role in the degradation process of plastic.


INTRODUCTION
Indonesia is facing plastic waste threats which currently reach 2.4 million tonnes and spread to aquatic environments.These plastics contaminate the natural environment and also endanger the ecosystem, especially when they degrade into small pieces (Moore, 2008).The plastic debris in the environment includes ropes, plastic bag or packaging, whichj are available in the form of macroplastics and microplastics.Large plastic items (macroplastics) have been indicated in the marine system for long time period (Derraik, 2002).Microplastics (< 5 mm) have recently attracted attention because of their ingestbility by organisms as well as transport for the pollutants into food chain (Teuten et al., 2009).Microplastics in the aquatic environment that can originate from various sources such as UV degradation and fragmentation of plastics or caused by damage during the transportation process, mechanical damage and also aquatic environment direct release (Andrady, 2003;Cole et al., 2011;Erikson et al., 2013;Rezania et al., 2018).
The most common and abundant polymers are high-density polyethylene (HDPE), lowdensity polyethylene (LDPE), polyvinyl chloride (PVC), polystyrene (PS), polypropylene (PP) and polyethylene terephthalate (PET), and among these plastic, PET and PP are the major plastic wastes which have different chemical-physical properties and functions (Andrady and Neal, 2009).PET and PP are derived from thermoplastic polymers which are mostly used for packaging materials.For example, single use plastic bottles for mineral water and soft drinks are made from polyethylene terephthalate (PET) which is highly recyclable with a density of 1.41 g / cm 3 (Mortula, 2013).Furthermore, PET contains two hydroxyl (OH) groups and dicarboxylic aromatic acid, which comprises a large, six-sided carbon (or aromatic) ring and two carboxyl (CO 2 H) groups.Under heat and catalysts, the hydroxyl and carboxyl groups will react to form ester (CO-O) (Venkatachalam et al., 2012;Farzi et al., 2019).The presence of a large aromatic ring in PET makes it stiff, strong, tough, hydrolytic, solvent resistant (Venkatachalam et al., 2012) and hygroscopic (Ma and Buhshan, 2003; CWC Best Practices in PET Recycling, 1997).In turn, polypropylene is polyolefin with linear hydrocarbon polymers (Arutchelvi, 2008) has the lowest density among commodity plastics that is about 0.94 g / cm 3  (Howe, 1999;Cole, 2002 andSchimanski, 2018).PP has three stereo configurations that can be distinguished, namely the isotactic, the syndiotactic and the atactic form.The isotactic PP (i-PP) contributes the most to the consumption of polypropylene because of its properties: ease to handle, stablility in aqueous solutions and organic solvents and also thermal-stability (Schimanski, 2018;Bertin et al., 2010).PP also has an excellent property regarding on chemical and temperature resistance which makes PP particularly suitable for application in many food packaging purposes especially that have to be sterilized frequently (Maddah, 2016).
The degradation process is the most important fate of plastics in the environment.Degradation of plastics may occur through some mechanisms that involve thermal, chemical, photo and biological degradation (Alshehrei, 2017;Gewert et al., 2015).Biodegradation of plastics occurs due to the action of extra cellular enzymes secreted by the microorganisms when the organisms attach to the surface of plastics leading to physical and chemical change of the latter (Lucas et al., 2008;Alshehrei, 2017;Arutchelvi et al., 2008).The growth of microorganisms utilising the plastics polymer as a carbon source and with the availability of oxygen, plastics will degrade completely by using CO 2 and the biomass as an ultimate product (Shah et al., 2008;Muthukumar and Veerappapillai, 2015;Arutchelvi et al., 2008;Arkatkar et al., 2009).
The hydrophilicity of plastic has important role in attaching microorganism cell to the plastic surface and therefore will affect the biodegradation process of plastic.PP which has a CH 2 group will be prone to attaching to hydrophobic polymeric surfaces (Arutchelvi et al., 2008).Due to the presence of ester and terephthalate group in PET, this plastic it polar molecule and therefore it is more hydrophilic (Venkatachalam et al., 2012).On other hand, Lucas et al. (2008) assumed that the esters group in PET makes it more resistant to biodegradation compared to other polymers.The extracellular polymeric substances produced by microorganisms can play a role as surfactants contain both hydrophobic and hydrophilic groups that support the exchanges between hydrophilic and hydrophobic phases (Lucas et al., 2008).
Microalgae are photosynthetic microorganisms which utilize CO 2 as carbon source to form biomass. Microalgae are mostly used as food sources in which they contain hydrocarbon, lipids and protein and other high added value compounds (Habib, 2008 andHariyati, 2008).Since they are used for food products, microalgae must be free from pollutants, including microplastics.Plastic can be used as carbon sources for microalgae and the release of plastic additives promote the growth of microorganisms by supplying the nutrient source (Rummel et al., 2017).Since micro-plastics have sizes of 1 to 5 mm (Lee et al. 2013), they are a suitable medium for bacteria and microalgae to form a bio-fouling.Bio-fouling that occurs on micro plastic surfaces causes shading effects which decrease light intensity, thus affecting the photosynthesis of microalgae (Sjollema et al., 2015;Yurtsever et al., 2017).Furthermore, the impact of micro plastics on microalgae growth is also affected by the availability of oxygen and CO 2 evolution as the consequence of plastic degradation (Shah et al., 2008;Arutchelvi et al., 2008).
The study on the biodegradation of PET and PP carried out simultaneously has not been found by the authors but some reports about PET and PP biodegradation in separate experiment have been observed.Farzi et al., (2019) studied the kinetic modelling in the process polyethylene terephthalate biodegradation waste using Streptomyces sp.The results showed that particle sizes and time of reaction were the most important parameters affecting biodegradation and the Michaelis-menten model can predict precisely the experimental results.Auta et al. (2018) evaluated the growth kinetics and biodeterioration of polyethylene microplastics by Bacillus sp and Rhodococcus sp and concluded that these microorganisms could modify and utilize PP microplastics as carbon source.
Lagarde et al. (2016) investigated the effect of polymer type during the interaction between micro plastic and freshwater microalgae.They reported that microalgae were over-expressed of sugar biosynthesis in HDPE rather than in PP.Furthermore, Yoshida et al., (2016) isolated a bacterium to breakdown the PET within 6 weeks.Long et al. (2015) evaluated the interaction between microplastics and phytoplankton aggregates.The result showed that marine aggregates can be an efficient sink for the microplastics.
Sharon and Sharon (2012) studied the plastic biodegradation of polyethylene terephthalate plastic in microbial culture and the degradation was slow and weak.It also demonstrated that microbes could act on the polyethylene terephthalate to form biofouling. Nowak et al. (2011) studied the biodegradation of modified PET by using polyester in Penicillium funiculosum culture.The result showed that modified PET was not significantly degraded in the presence of the culture.Since the interaction of micro plastic and microorganism is relatively strong, further research is required.The purposes of this research were to evaluate the contribution of Spirulina sp in the process of plastic degradation and to observe the impact of microplastics on the Spirulina sp growth.

Materials
Polyethylene Terephthalate (PET) used in this research was obtained from Danone Indonesia that already standardized by SNI 19-4370-2004 (Nasional Indonesian Standart for Plastic bottle Single-use).Polypropylene (PP) 15 μm thick was provided by PT.Indofood Sukses Makmur Tbk.

Spirulina sp cultivation
Spirulina sp used in this experiment was supplied by C-Biore Laboratory, Diponegoro University.The experiment was started by testing the physiological form of Spirulina sp which included maximum cell density by using a spectrophotometer, measurement of pH, temperature, oxygen content and CO 2 content.Furthermore, Spirulina sp is placed in 2 pieces of 35 cm x 25 cm glass ponds and 7 pieces of 500 mL Erlenmeyer, each equipped with an aerator as a source of oxygen and LED lights (3000 lux) to provide light intensity, then the temperature is maintained at 24-26 o C and pH between 7-8.Nutrition needed to maintain the Spirulina sp growth is given every 2 days with a mix of TSP and Urea 12.5 mg / 250 mL of Spirulina sp and 100 mg NaHCO 3 / 250 mL Spirulina sp (Hadiyanto and Azim, 2012).

Sample Preparation
PET and PP plastics were washed with etanol and dried at room temperature for 24 hours and then the plastic was cut to the size of 5 x 5 cm to be applied in a glass ponds containing Spirulina sp.Microplastics were obtained by cutting a PET and PP plastic in the same size between 1-2 μm.The microplastic was weighted carefully at 150 mg, 200 mg and 275 mg and mixed into 500 mL of Spirulina sp culture.Stirring was done with the aerators so that microplastics were distributed into the Spirulina sp culture properly.Nutrient, pH, temperature, light intensity and oxygen supply were maintained and the growth was measured by using a spectrophotometer (Spectroquant pharo 300 M and Spectrophotometer SP -300 Optima) at 560 nm wavelength and ultrapure water was used as a blank solution (Hadiyanto et al., 2012).

Tensile strength measurement
In SNI 7818: 2014 and SNI 7188.7.2011, it was stated that one of the degradability tests for plastic is by using tensile strength test and it was also supported by ISO 527-3 (Lucas et

Morphologies
The morphology of PET and PP was observed using scanning electron microscope (SEM) and combination with Energy Dispersive X-ray spectroscopy (EDX) to determine the inorganic elements contained in the material (Lucas et al., 2008).The analysis was conducted at room temperature and metalized using Au.A Jeol (model JSM-6510 LA ) at 3000x magnification.

FTIR analysis
FTIR is a common technique used for the study of macromolecules such as PET and PP polymer that was recommended for investigation of plastic degradation as mentioned in ISO 4582 and ISO 4892 for UV exposure, and for microorganism's surface colonization in ISO 846 and ISO 11266 (Lucas et al., 2008, Melissa et al., 2018, Schmitt et al., 1998).PET and PP plastics that were applied in Spirulina sp were taken every 7 days for about 112 days.Prior to the FTIR test, plastics were rinsed with aquadest and left to dry for 24 hours, then the plastic was cut at a size of 1.5 x 2 cm.A Perkin Elmer Type Frontier was used to collect spectra from 4000-200 cm -1 (SNI 19-4370-2004 method) and ASTM D6288-89.FTIR test was also conducted in Spirulina sp which had interacted with microplastic treatment for 7 days.Filter Spirulina sp containing micro plastic with diameter of 200 mm / 8 "stainless steel 40 mesh sifter sieve fine wire strainer to obtain Spirulina sp without microplastic.

Contribution of Spirulina sp in the plastic degradation processes
The standard test of the elasticity (elongation) properties for degraded plastics in Indonesia is regulated by SNI 7188.7:2011 (BSN, 2011) which requires tensile elongation, while on an international scale, it is regulated in ASTM D3826 concerning the procedures for determining the end point of degradation in Polyethylene and Polypropylene plastic using tensile test.The quantitative relationship between tensile strength and degradation is the first step in the process of investigating yjr plastic degradation (Guo et al., 2016).When plastic changes both due to biotic and abiotic factors, the strength of stress and the versatility of plastic will alter in line with changes in the molecular structure of the polymer.Therefore, the initial identification of the plastic degradation process is by measuring the tensile strength.There are several factors that influence the tensile strength of plastic, namely the molecular structure that also affects the density of the plastic, the temperature at which the plastic is applied and the chemical composition of the plastic itself.Measurements of tensile strength were carried out on two types of plastic and the measurement results can be shown in the curve of tensile strength degradation over time.

The change in mechanical polymer property by tensile strength measurement
Figure 1 depicts the tensile strength degradation over time of two plastics (PP and PET).The decrease in tensile strength in PET is far greater than the rate of decrease in tensile strength of PP.The density of PET is higher (1.37 kg / m 3 ) than PP density (0.94 kg / m 3 ), which leads to higher tensile strength value.
The tensile strength of both plastic during interaction with Spirulina sp for 112 days showed a greater rate of tensile strength decrease of PET compared to PP.The hydrophilic and hydrophobic nature of polymers plays a role in biofilm and hetero-aggregation formations in the biodegradation process of polymers in the aquatic system (Merina, 2014; Cerca et al., 2005;Lobelle et al., 2011).The formation of biofilms by microorganisms can improve the hydrophilic properties of the plastic surface (Lobelle et al., 2011).PET is more hydrophilic than PP, as it contains polar group (C = O bond) (Lai et al., 2006).Therefore, PET has greater potency to experience exopolysaccharide (EPS) heteroaggregation produced by Spirulina sp.Spirulina sp has both hydrophilic and hydrophobic properties because of its high protein contents (Bashir et al., 2015).The exopolysaccharide heteroaggregation that occurs in PET surfaces happened when microalgae reach the stationary growth phase (Long et al., 2017) where (EPS).Heteroaggregation is very important in determining the fate, transportation, transformation and toxicity of nanoparticles in aqua phase (Wang et al., 2015).Heteroaggregation in PET results in more brittle PET which causes the tensile strength of PET decrease faster than in PP.
The difference in the decrease rate of tensile strength in PET and PP can also be caused by chemical composition, especially the amount of additives added in the process of plastics production (Lagarde et al., 2016).

The change of organic groups investigation by FTIR Analysis
FTIR analysis is a suitable technique for investigating the degradation of plastics in the environment through the determination of changes in various functional groups that contribute to the plastic polymer compound (Loakeimidis et al., 2016).Some results of research on the FTIR's ability in analyzing organic functional groups in plastic polymers were reported in a review of Jung et al., (2018) who concluded that FTIR spectroscopy is able to provide a simple, effective and non-destructive method for identifying and distinguishing functional groups organics widely in most plastic polymers with high accuracy results.
Figure 2 shows the FTIR analysis of PET before and after interaction with Spirulina sp.The characteristic of PET were identified by its functional groups of for C = O stretch (ketone) at wavelength of 1718 cm -1 , C=C aromatics at 1505 cm -1 , 1523 cm -1 , 1578 cm -1 and 1613 cm -1 , (CO) aliphatic ether at 1125 cm -1 , aromatics (CH) at 874.5 cm -1 and aromatic bonds (CH) at 733 cm -1 .
The spectrum of PET after interaction with Spirulina sp is characterized by the decreasing the peak intensities of the band located at 1613-1505 cm -1 (aromatic C=C) (Fig. 2).It also shows there is no progressive reduction in the relative intensity of the peak carbonyl and the appearance of new absorption bands was observed.
Figure 3 shows the FTIR-ATR spectrogram of the PP surface layer before and after interaction with Spirulina sp.Before the interaction with Spirulina sp, the results of the PP spectrogram control gave an important peak in the wavelength region of 3100-3700 cm -1 (water OH stretch, 1456 cm -1 (CH 2 bend), 1377 cm -1 (CH 3 bend), 1166 cm -1 (CH bend, CH 3 rock, CC stretch), 997 cm -1 (CH 3 rock, CH 3 bend, CH bend), 840 cm -1 (CH 2 rock, C-CH 3 stretch), 809 cm -1 (CH 2 rock, CC stretch).After interaction with Spirulina sp, the appearance of absorption bands located at 1599 and 1534 cm -1 corresponds to Amide (C=O) and very strong peak at 1731, 48 cm -1 that corresponds to an ester and keton (C=O).Domagala (2012) found a new absorption band within the wave number range of (1730-1680)cm -1 that corresponds to a carbonyl group as a results of the nucleophilic substitution of PP.
Moreover, the absorption band of carbonyl group in the PP spectrum is broad which indicates the presence of carbonyl group in various products of oxidation such as aldehydes and ketons (Carlsson and Wiles 1969).Figure 3 also shows the new peak at 3343 cm -1 , which reveals the presence of Spirulina sp as it was also reported by Theivandran et al., (2015).The results of FTIR-ATR Spectrogram of PP after interaction with Spirulina indicate a particular activity of oxidative degradation process of PP in Spirulina sp medium.

Morphological evaluation of microplastics using SEM/EDX Analysis
The SEM analysis was conducted to investigate the changes in the surface morphology of the plastics.Nauendorf et al., (2016) proved in his study that biofilm formation in surface of plastic depends on several factors such as plastic surface roughness, plastic hydrophilic surface properties, the properties of substratum and also the bulk liquid properties as well as on cell surface properties.Figure 4 shows the SEM micrographs of the PET and PP surfaces before and after 112 days of interaction with Spirulina sp.Before interaction with Spirulina sp (Fig. 4a) the PET samples had a smooth area among the blisters and (Fig. 4c) the PP samples had a coarser and more textured surface with defects.However, after incubation with the Spirulina sp, surface erosion and the formation of pits and cavities on the surface of the samples were observed (Figs.4b and 4d).The  presence of pits and cavities may be because of the absence of biofilm that become the areas colonized by microorganism, also suggesting that the fungus penetrated into the PET and PP matrix and a bacterial biofilm formed on the surface of plastics.Nakkabi et al.(2015) found that Bacillus subtilis strain has an effect on the change of PET surface morphological heterogenecity and signs of erosion which show the ability to degrade the PET.This experiment revealed that PET and PP also contribute a carbon source for Spirulina sp to form carbon dioxide as one of the major metabolic end products under aerobic conditions (Shah et al., 2008).In ISO 14852, it was depicted that the identified ultimate aerobic biodegradability of plastic materials in an aqueous medium can be performed by analysing the evolved CO 2 and this can be used as a reference in considering the amount of carbon from EDX investigation on PET and PP before and after interacting with microalgae.
Table 1 shows a decrease in carbon concentration by 48.61% in PET while in PP there is a carbon decline of 36.7%.The plastics that contain chemical compounds in the manufacturing process also have the ability to release and distribute contaminants to the environment as well as contaminate the environment by harmful chemical pollutants, and are able to absorb contaminants from the environment (Teuten et al., 2009).Rummel et al., (2017) reported the occurrence of chemical pollutants transport through the biofilms formed on the surface of the plastic.This is evident from the results of EDX that after plastic treatment with Spirulina sp, several inorganic elements were newly identified in plastics.These inorganic elements can be derived from the nutrients added to Spirulina sp media and from the release of additive compounds from the plastic which are added during the process of making plastic itself.

Optical density measurement for Spirulina sp growth
In this part of the experiment, we characterized the impact of PET and PP microplastics to Spirulina sp growth by measuring the optical density of Spirulina sp in various concentrations of microplastics (Fig. 5).
The decreasing microalgae growth is statistically significant (Fig. 6  after 72 hours in the presence of 250 g/L of polystyrene.However, exposing different microplastics concentrations level gives different microalgae growth rates whereas the higher microplastics concentration in the microalgae led to lowering the growth rate of the microalgae.Microalgae with the addition of PET and PP (Figure 8), generally have lower growth rate constant as compared to the microalgae without microplastic addition (0.399 day -1 ).This is because of the presence of microplastics in Spirulina sp culture may cause shading effects and lead to the inhibition of light intensity which is important in the process of microalgae  photosynthesis (Hadiyanto et al., 2012).The microplastics dosage applied with a concentration of 150 mg/mL results in higher microalgae growth rates in PET (0.3584 day -1 ) compared to PP (0.2647 day -1 ) whereas at higher concentrations of 200 mg/mL and 275 mg/mL the opposite results were found in which the microalgae with the addition of PP microplastics (0.1229 day -1 and 0.907 day -1 ) have a higher growth rate than the microalgae with the addition of PET microplastics (0.1144 day -1 and 0.0832 day -1 ).

FTIR analysis
The FTIR analysis of Spirulina sp in fresh water without any treatment (Fig. 7 and Fig. 8), represent the following associated functional groups: at the wavelength of 3572 cm -1 representing the O-H stretching vibration and thus presence of alcohols and phenols.The peak at 3436 cm -1 represents the strong N-H (amine) and then at 1651 indicates -C=C-stretching vibration presence of alkenes, and peak of 1271 cm -1 presence of C-H wag (-CH 2 X) stretching of alkyl halides.Furthermore, the frequency ranges of 1200-1020 cm -1 is the presence of C-OH stretching and peak of 838 cm -1 present N-H symmetric stretching vibration primary of secondary amines.The last peak at 618 cm -1 representing C-Br stretching vibration indicates the presence of alkyl halides compounds.Some new peaks emerge and change of their intensity after additions of microplastics in Spirulina sp culture.The appearance of the absorption spectra in the region of 2300 -2400 cm -1 only occurs in PET under microplastics treatment.The highest concentration of PET was shown by the higher intensity of the spectra in that region (from 150 mg/500mL to 275 mg/500 mL), and this is slightly different to the PP application.The appearance of spectra in the 2300-2400 cm -1 region only occurs in Spirulina sp by applying 200 mg/500 mL and 275 mg/500 mL of microplastics.Gerakines et al. (1995) reported the appearance spectra at peak 2343 cm -1 , which was identified as the existence of CO 2 .The fact that higher CO 2 intensity in Spirulina sp added by PET and PP is related to the availability of carbon supplied by microplastic which lead high amount to be converted to CO 2 .This conversion process is part of the mineralization in the biodegradation process.Gupta et al. (2007) explained that the mineralization process in plastic biodegradation will occur in fragmented plastics where the microplastic residues produced are carbon as food sources and the converted energy produces CO 2 .Furthermore, Shah et al., (2008) reported that under aerobic conditions oxygen was used by microbes to oxidize carbon to produce CO 2 as a major metabolic end product and this is also sup- The decrease of intensity in some peaks and the emergence of new peaks after application of PET and PP in Spirulina sp is an indication of the interaction between microplastics and Spirulina sp.The appearance of FTIR absorption peak in the region of 3700-3800 cm -1 in Spirulina sp under addition of both microplastics is in line with the increasing microplastics dosage applied.The absorption is due to O−H stretching modes in the range of 3800-3000 cm -1 .The IR spectra can be represented as a sum of contributions from interfacial and bulk like water (Profio et al., 1998)

CONCLUSION
In this study, the measurements of tensile strength, analysis with FTIR-ATR and SEM-EDX were carried out on PET and PP after interacting with Spirulina sp for 112 days to understand the biodegradation process in PET and PP.From the measurement results of tensile strength, PET appears to provide a greater decrease in tensile strength compared to PP, but the opposite condition occurs in FTIR-ATR analysis, which shows a more significant change in functional groups in PP compared to PET.Furthermore, both PET and PP surface imaging with SEM after interaction with Spirulina sp in 112 days showed signs of surface alteration.Although the signs of biodegradation in PET and PP are indicated by the results of the analysis, but it still cannot be concluded that the process of biodegradation with microalgae provides the most effective results.Further research is required to obtain more infromation on the effectivity of microalgae Spirulina sp on its involved in biodegradation processes.Another study of PET plastic and PP impact to Spirulina sp growth shows a strong influence of PET and PP on the growth of Spirulina sp.The interaction between plastic and microalgae provides phenomena that need to be further studied to devise a solution in handling the abundance of plastic waste in the aquatic system and maintaining the survival of organisms in waters.

Fig. 1 .
Fig. 1.The changes of tensile strength of polyethylene terephthalate and polypropylene upon degradation time under Spirulina sp influences

Fig. 2 .
Fig. 2. The FTIR comparative spectra of the PET : (a) before treatment and (b) after treatment with Spirulina sp.

Fig. 3 .
Fig. 3. FTIR comparative spectra of the PP: (a) before treatment and (b) after treatment with Spirulina sp.
) among PET, PP and control during 7 days cultivation.Lagarde et al. (2016) found the decreasing microalgae growth after 78 hrs contact and Besseling et al., (2016) resulted in the decrease of microalgae growth

Fig. 4 .
Fig. 4. Morphological analysis using SEM in magnification x 1000 visualizations of the surface topography and roughness of the (a)PET without treatment, (b) PET after treatment with Spirulina sp, (c) PP without treatment, (d)PP after treatment with Spirulina sp for about 112 days.

Fig. 6 .Fig. 7 .
Fig. 6.Comparison of effect of microplastics in various concentrations to Spirulina sp growth rate (μ) ported by the research of Hoffmann et al. (1997) and Lucas et al. (2008).Arutchelvi et al. (2008) and Rummel et al. (2017) explained that biomass accumulation characterized by the growth of microorganisms was capable to utilize polymers as a carbon sources and this cause the main chain cleaves that leading the formation of low molecular weight compounds as impact of the extra cellular enzymes secreted by the microorganism, called as bio-fragmentation.This step is followed by diffusing oligomers into the microorganism to obtain assimilation.When the biodegradation is accomplished, it will produce CO 2 , H 2 O and biomass under aerobic conditions.

Fig. 8 .
Fig. 8. FTIR comparative of spectra of Spirulina sp under micro plastic Polypropylene influence in various concentrations.
al., 2008, Alvarez-zeferino et al., 2015, Guo Meng et al., 2016, Hoffmann et al., 1994, Hongliang et al., 2017, Strapasson et al., 2005).Tensile strength tests were conducted by tensile meter (Brookfield CT3 -4500) which were carried out on plastic before and after treatment by Spirulina sp.The plastics exposed by Spirulina sp treatment were measured every 7 days for 112 days to measure their tensile strength.

Table 1 .
The EDX result for PET and PP before and after treatment with Spirulina sp