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Food Sci Anim Resour. 2020 Apr; 40(3): 311–322.
Published online 2020 Apr 30. doi:10.5851/kosfa.2019.e79
PMCID: PMC7207083
PMID: 32426712
Zubair Hussain,1 Xin Li,1 Muawuz Ijaz,1 Xiong Xiao,1 Chengli Hou,1 Xiaochun Zheng,1 Chi Ren,1 and Dequan Zhang1,*
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Abstract
This study was undertaken to evaluate the impact of Chinese cinnamon powder(w/w), at the levels of 0.5%, 1.5%, and 2.5% and control(without additive) on ground lamb meat quality. The samples were stored at4°C and examined for pH, color, lipid oxidation (thiobarbituric acidreactive substances) and total viable counts (TVC). The results demonstratedthat pH values were declined with the increase of Chinese cinnamon levelscompared to control group. The L* values throughout the storage weresignificantly higher (p<0.05) in the control group than in othertreatment groups, while a* values were decreased with the increase of Chinesecinnamon levels. The addition of Chinese cinnamon powder strongly inhibited(p<0.05) thiobarbituric acid reactive substances (TBARS) and TVC in alltreated samples. It can be concluded that Chinese cinnamon powder in lowerconcentration 0.5% has the ability to maintain the quality of ground lambin comparison with other treated samples.
Keywords: lamb, Chinese cinnamon, meat quality, storage
Introduction
Seasoning of meat is a possible solution to enhance the color stability, minimizelipid oxidation and improve microbial safety of meat products. The seasoned meatproducts are more susceptible to spoilage of microorganism (Rysman et al., 2016). These products are usually marketed atrefrigerated temperature (2°C−5°C) in order to increase shelflife (Radha Krishnan et al., 2015). Thespoilage due to the lipid oxidation has deleterious consequences on the quality offresh meat and other meat products resulting in massive economic losses (Shahidi and Zhong, 2010). While microbialactivities in the food products may also destroy the quality of meat by thedevelopment of undesirable responses involving the worsening of odour, color andtextural properties of food products (Lucera et al.,2012).
The spoilage factors in meat products sometimes produce toxic materials which arehazardous for human health (Jiang and Xiong,2016). Therefore a vast amount of antioxidants, which have beenchemically synthesized, are added to the meat products, such as butylatedhydroxyanisole (BHA) and butylated hydroxytoluene (BHT). However, use of potentiallycarcinogenic substances is not permitted in some countries (Falowo et al., 2014). Therefore, meat industry is largelyapplying antioxidants from plant sources as safer alternatives and preservatives(Devatkal and Naveena, 2010).
A number of plant materials have been used directly or indirectly, as seasoningmaterials for the antimicrobial purpose, to improve the meat quality (Appendini and Hotchkiss, 2002). The naturalpreservatives used for the meat producers include cinnamon, clove, rosemary, basil,thyme, oregano, lemon leaf, ginger, basilica, balm, coriander and many of them aregenerally recognized as safe (GRAS) in food industry (Alfonzo et al., 2017; Jiang and Xiong,2016; Khaleque et al., 2016).Chinese cinnamon (Cinnamomum cassia) is usually add to the deserts,drinks or bakery products, majorly in South Asian and Central Asian regions, andconsidered as a vital spice in many meat products. Cinnamon is commonly used in meatand fast food products for culinary purpose, and has been usually described as apowerful antioxidant and antibacterial agent in the seasoned meat and fish products(Al Sahlany, 2017; Ozogul et al., 2017). Previous study has been confirmed thatcinnamon has effective free radical scavenging activity due to the presence ofbioactive compounds (Radha Krishnan at al.,2014). Cinnamaldehyde is the major constituent responsible for it highantioxidant activity (Dudonne et al., 2009)and the ability of cinnamon to preserve meat quality has been tested in beef burger,grass crabs and inactivate Listeria monocytogens in ground beef andchicken meatballs (Ghabraie et al., 2016;Huang et al., 2017; Khaleque et al., 2016).
The effects of adding Chinese cinnamon powder directly into the ground lamb meat tomaintain meat quality has not been tested previously. Therefore the aim of thisstudy was to examine the effectiveness of different concentrations of Chinesecinnamon powder on the ground lamb meat quality. For this purpose, concentrations of0.5%, 1.5%, and 2.5% Chinese cinnamon powder were added to theground lamb meat and total viable counts (TVC), pH, lipid oxidation and colorattributes were evaluated. Furthermore, the antioxidant activity of Chinese cinnamonaqueous extract was also measured. This study may provide potential implications insubstituting synthetic antioxidants with natural ones.
Materials and Methods
Materials and preparation of extracts
Dried bark of Chinese cinnamon was obtained from a local supermarket. Chinesecinnamon bark was divided into small pieces and grounded to powder by a highperformance kitchen grinder (High Speed Universal Grinder, Tianjin, China). Thepowder was sieved with the help of sieve (1.651 mm, ASTM No. 10), and packedinto 100 g packs and then stored in high-density polyethylene bags at25°C until its further usage. Exactly 50 g powder was refluxed by using450 mL distilled water for 5 h in enclosed flasks with constant shaking at180×g by following the method from Sivarajan et al. (2017) to obtain a 10% w/v water extract.Then the extract was cooled and filtered twice using Whatman filter paper andused for performing the DPPH antioxidant assay.
Antioxidant activity
The DPPH-radical-scavenging activity of aqueous extracts was assessed byfollowing the protocol of Elahi and Mu(2017). Chinese cinnamon and BHT at concentrations of (20, 40, 60mg/mL) were prepared by addition of 2 mL of freshly prepared DPPH solution (0.1mM in 95% methanol). The solution was vortexes using a mixer andincubated in dark at 27°C for 40 min. After that, each sample absorbancewas checked at 516 nm using a UV spectrophotometer (Shimadzu UV-1800Spectrophotometer, Kyoto, Japan) at room temperature. The radical-scavengingactivity of samples were calculated in to percentage by following equation.
where “AE” showed the absorbance of solution at 516 nm after mixingthe 1 mL of all samples with 2 mL of 0.1 mM DPPH solutions and 30 min incubationat room temperature, while “AD” showed the absorbance of 2 mL 0.1mM DPPH solutions mixed with 1 mL Milli-Q water.
Sample preparation, packaging and storage
Fresh lamb meat (Oyster muscles) was purchased from a localmarket at 24 h post-mortem and was placed in insulated polystyrene ice boxes andtransferred to the laboratory within 1 h. The muscles were trimmed to removeconnective tissues under hygienic conditions and minced using a mincer machine(8 mm plates). The minced lamb meat samples were assigned to the following fourtreatments: control (without any additive), 0.5%, 1.5%, and2.5% of Chinese cinnamon powder applied on the 100 g ground lamb meat.Immediately after adding the Chinese cinnamon powder, samples were thoroughlyhand-mixed using a bowl mixer and 100 g ground lamb meat (round shape and 1.5 cmthick) was prepared for each treatment with three replicates. The lamb meatsamples were vacuum packaged (VP), properly labelled and stored at 4°Cfor 16 days. The vacuum packaged samples were sealed in polyethylene bags (20/70mm) (Vacioplast, Salamanca, Spain) with an oxygen permeability lower than 40cm3/(m2 day atm). After that, the samples werecollected at 0 (approximately 24 h post mortem), 4, 8, 12 and 16 days ofstorage. At the time of sampling, 10 g sample was collected immediately underaseptic conditions for microbiological analysis. While the meat color wasdetermined after blooming for 30 min at 4°C and then the remainingsamples were frozen at −80°C priors to thiobarbituric acidreactive substances (TBARS) analysis. The above experiment was carried out intriplicates.
Analysis of meat samples
pH
The pH value of ground lamb was determined by pH meter (Testo 205 pH meter,Lenzkirch, Germany). Before measurement, pH meter was calibrated by buffersof different pH concentrations i.e. 4.00 and 7.00 at 25°C. The glassrod of pH meter was inserted directly into the ground meat sample. Eachtime, four readings were recorded from different locations and averaged.
Color
The meat surface color was recorded using Minolta spectrophotometer (CM-600d,Konica Minolta Sensing Inc., Osaka, Japan) with 8 mm diameter measuringaperture size Illuminant D65, 10° standard observer and CIE L*, a*,b* color score. Four measurements were recorded throughout the surface ofselected samples (Li et al., 2017).The color of meat samples at 0 days was taken after collecting the fresh-cutsurface from the local market. At each time point the vacuumed packedsamples were opened and the surface color of ground lamb meat was measuredat 0, 4, 8, 12, and 16 days after 30 min of blooming at 4°C.
Lipid oxidation
Lipid oxidation analysis was conducted with minor modification by followingthe method described by Belles et al.(2017). Briefly, 20 mL of 10% tricloracetic acid (VWR) wasmixed with 10 g of meat and hom*ogenized at 724×g by using ultraturraxfor 90 s (T-10 basic, IKA-Werke, Staufen, Germany). Then the samples werecentrifuged for 30 min at 10°C at 2,897×g (High-SpeedRefrigerated Centrifuge, CR 21N, Hitachi, Tokyo, Japan). The supernatant wasfiltered by using filter paper and 2 mL of the filtrate was added with theequal quantity of TBA 20 mM (Sigma-Aldrich, St. Louis, MO, USA). After that,the mixture was vortexed and incubated in a water-bath at 97°C for 20min. At the end, samples were cooled under tap water at ambient temperatureof 15°C and absorbance was measured at 532 nm by usingspectrophotometer (Shimadzu UV-1800 Spectrophotometer, Kyoto, Japan). TheTBARS, mainly malondialdehyde (MDA), was calculated from a standard curve of1,1,3,3-tetraethoxypropane (TEP; Sigma-Aldrich), the lipid oxidation wasexpressed as the average of three replicates per sample in mgmalondialdehyde/kg meat.
Microbiological analysis
TVC were inspected based on standard plate count method by the method ofZhang et al. (2016). In brief, 10g sample was taken aseptically from vacuumed bags and hom*ogenized in 90 mLof sterile physiological saline in stomacher bags for 1 minute. Formicrobial enumeration, suitable serial dilutions were prepared using thesame diluent, by following the protocol of International Organization forStandardization’s (ISO, 2003).Then the deriving suspension was serially diluted (1:10) in sterilephysiological saline water and 1 mL samples of appropriate dilutions werepoured into the petridishes, containing 15–20 mL of plates count agar(PCA). The number of bacterial colonies on the plates were enumerated afterincubation for 48 h at 37°C and expressed as Log CFU (colony formingunits)/g meat.
Statistical analysis
The experiment was designed with Chinese cinnamon treatments and storage times asfixed factors and replicates as random factor. General linear model (GLM) wasused to express the significance of differences (p<0.05) between means.Statistical analysis of data was performed using the IBM statistical package forsocial sciences (SPSS) Statistics 22 software (SPSS Inc., Chicago, IL, USA).Duncan multiple range test was applied to determine the significant difference(p<0.05). The data were expressed as the mean±SD. Experiments werereplicated three times and all parametric measurements were carried out induplicate.
Results and Discussion
Antioxidant activity
The DPPH radical scavenging activity of Chinese cinnamon aqueous extract at theconcentrations of 20, 40, and 60 mg/mL were considerably lower than the pureantioxidant BHT as shown in Fig. 1(p<0.05). The Chinese cinnamon aqueous extract showed strong antioxidantactivity in all concentrations but the highest antioxidant activity was seen in60 mg/mL indicating the highest radical scavenging activity (p<0.05).
Fig. 1.
The DPPH radical scavenging activity (%) of aqueous extract ofChinese cinnamon powder and BHT.
A,B Different letters indicated a significant difference(p<0.05) between the same concentrations of Chinese cinnamonaqueous extract and BHT. BHT, butylated hydroxytoluene.
Previously reported studies have confirmed that cinnamon is distinguished by itseffective radical scavenging activity due to bioactive substances (Radha Krishnan et al., 2014). The higherantioxidant activity in cinnamon might be due to the presence of significantamount of phenolic antioxidants and flavonoids compound (Jayaprakasha et al., 2007). Kuspradini et al. (2016) reported that cinnamon aqueous extract aswell as cinnamon oil, have shown considerable antioxidant activity. Radha Krishnan et al. (2015) claimed thatcinnamaldehyde was the responsible compound in the cinnamon for its highantioxidant activity.
pH
The results of adding Chinese cinnamon powder on the pH of ground lamb meatstored at 4°C were presented in Table1. The pH of control had a significant increased from day 12 to 16while that of all other treatments samples remained the same during storage. ThepH for all samples was same at day 0, while at day 4 the pH value of sampleswith 1.5% Chinese cinnamon powder was significantly lower (p<0.05)that of the control group and the other treatment groups. The pH values ofsamples from the control group and the 0.5% treated group had nosignificant difference with each other on day 8, whereas the stored samples withconcentrations of 1.5% and 2.5% Chinese cinnamon powder showed asignificant difference (p<0.05) compared with control group. With theincrease of storage time, the samples with 2.5% concentration showedlower pH values at day 12 and 16 than control and other treated samples(p<0.05). Overall, the control group had a higher pH (p<0.05) atday 12 and 16 than that of other treatments during the storage.
Table 1.
Effect of different concentrations of Chinese cinnamon powder on thepH of vacuumed ground lamb meat at 4°C
| Treatments (%) | Storage time (days) | ||||
|---|---|---|---|---|---|
| 0 | 4 | 8 | 12 | 16 | |
| Control | 5.62±0.03Ab | 5.57±0.01ABbc | 5.46±0.01Ac | 5.67±0.03Ab | 5.80±0.03Aa |
| 0.5 | 5.64±0.02Aa | 5.54±0.02Bb | 5.40±0.03ABd | 5.45±0.04Bc | 5.48±0.03Bc |
| 1.5 | 5.62±0.03Aa | 5.49±0.03Cb | 5.42±0.04Bb | 5.42±0.03Bb | 5.48±0.03Bb |
| 2.5 | 5.60±0.04Aa | 5.60±0.04Aa | 5.36±0.03Bb | 5.38±0.01Cb | 5.35±0.04Cb |
Open in a separate window
Results are presented as means±SD (n=3).
A–DMean values in different capital letters within the same columndiffer significantly (p<0.05).
a–dMean values in different small letters within the same row differsignificantly (p<0.05).
The increment in the pH of sample stored under control group during the storagemight be due to the generation of some basic compounds and ammonia caused byproteolysis resulting from the growth of microorganism (Chaijan et al., 2005; Masniyom et al., 2002). RadhaKrishnan et al. (2014) and Brillianaet al. (2017) reported that during refrigerated storage, lower pH wasnoted in the raw beef meat samples treated with cinnamon oil compared tonon-additive control. In the present study, the lower pH in the treatmentsamples might be due to the strong activity of bioactive compounds from theChinese cinnamon powder.
Color
The L*, a*, and b* of Chinese cinnamon powder treated and untreated ground lambmeat samples stored at 4°C were presented in Table 2. L* (lightness) value of samples from control groupwas significantly (p<0.05) higher throughout the whole storage timecompared with other treatments. Among all Chinese cinnamon treated samples, thelower L* values were noted in samples with 2.5% concentration treatmentin all storage days except for day 8 (p<0.05). The addition of Chinesecinnamon powder directly into the ground lamb might decrease the lightness oftreated samples. The a* (redness) value of ground lamb was also affected by theaddition of Chinese cinnamon powder during storage as shown in Table 2. The a* value of the control groupshowed a decreasing trend throughout the storage as compared to all theexperiment groups. The initial a* values for all stored samples at 4°Cwere significantly different from each other (p<0.05). The a* values ofcontrol group and 0.5% treatment group were significantly higher at day 4than those of samples with 1.5% and 2.5% treatments. At day 8 thea* value of control samples was considerably higher (p<0.05) than treatedsamples with 0.5% and 2.5% of concentration, while no significantdifference was noted in samples from 1.5% treatment. As the daysproceeded, at day 12 there were no significant differences between control groupand all other Chinese cinnamon treated samples. The a* value for samples storedunder control was lower than that of 1.5% treated samples at day 16,while no significant difference was found between samples with 0.5% and2.5% of Chinese cinnamon powder (p<0.05).
Table 2.
Effect of different concentrations of Chinese cinnamon powder oninstrumental color parameters (CIE L*, a*, b*) of vacuumed ground lambmeat at 4°C
| Parameters | Treatments(%) | Storage time (days) | ||||
|---|---|---|---|---|---|---|
| 0 | 4 | 8 | 12 | 16 | ||
| L* (Lightness) | Control | 53.06±0.79Aa | 52.76±0.73Aab | 51.80±0.87Aab | 51.24±0.70Aab | 50.98±1.88Ab |
| 0.5 | 47.02±0.40Bb | 48.37±1.67Bab | 49.25±1.61Ba | 49.25±0.62Ba | 48.20±0.56Bab | |
| 1.5 | 45.77±0.72Cb | 45.11±0.94Cb | 44.44±0.77Cb | 47.39±0.99Ca | 46.20±1.80Bab | |
| 2.5 | 41.67±0.49Db | 42.03±0.79Dab | 43.55±0.88Ca | 42.31±0.90Dab | 41.76±1.68Cb | |
| a* (Redness) | Control | 16.87±0.35Aa | 15.42±0.73Aab | 14.06±1.05Ab | 13.86±1.12ABb | 12.20±0.66Bc |
| 0.5 | 15.63±0.48Ba | 14.81±0.47Aab | 13.42±0.81BCb | 13.08±0.75Bb | 13.21±0.70ABb | |
| 1.5 | 12.14±0.52Db | 12.21±0.89Bb | 13.37±0.38ABab | 14.16±0.57Aa | 13.64±1.09Aa | |
| 2.5 | 13.26±0.18Cab | 12.55±0.16Bab | 11.91±0.28Cb | 13.45±1.10ABa | 12.46±1.18ABab | |
| b* (Yellowness) | Control | 19.31±1.20Aa | 15.34±1.34Ac | 17.33±1.48Ab | 16.61±1.07Ab | 16.11±0.62Ab |
| 0.5 | 17.87±0.54ABa | 15.11±1.15Ab | 17.27±0.67Aa | 16.27±0.67Aa | 16.50±0.95Aab | |
| 1.5 | 17.31±1.02Ba | 14.66±0.73Ab | 16.29±0.59ABa | 16.36±0.78Aa | 16.50±0.31Aa | |
| 2.5 | 15.31±0.78Cb | 13.81±0.45Bc | 15.48±0.68Bb | 16.07±0.61Aa | 16.20±0.76Aab | |
Open in a separate window
Results are presented as means±SD (n=3).
A–DMean values in different letters within the same column differsignificantly (p<0.05).
a–dMean values in different letters within the same row are differsignificantly (p<0.05).
The b* (yellowness) value for control group was higher at the initial day thanthat of the 1.5% and 2.5% treated groups (p<0.05), while nosignificant difference was noted in stored samples for 1.5% treatment.Whereas, on day 4 and 8 the only significant difference was noticed betweencontrol and 2.5% treatment groups than the samples with 0.5% and1.5% treatment. The b* values for all stored samples included control andother Chinese cinnamon treated samples had no significant difference(p<0.05) after day 12 to 16 during storage.
It was proved that the meat discoloration was directly related with the storagelength, and a* had a negative correlation with storage time (Terns et al., 2011), which might bepossibly associated with an increase in TBARS (Grimsrud et al., 2008). Previous results confirmed that the additionof natural antioxidants might slow down the formation of metmyoglobin,ultimately delaying the deterioration of red color (Belles et al., 2017; Xia etal., 2009). Keokamnerd et al.(2008) observed that a* value was reduced in the minced chicken after12 days of storage. The lessening in the intensity of redness values duringstorage was probably due to the relationship between lipid oxidation and coloroxidation in the meat (Lynch and Faustman,2000). In the current study, the fluctuations in the a* values wereobserved in all samples during the storage. The results were in accordance withthe previous findings (Ozunlu et al.,2018; Zhang et al., 2016). Thevariation in the a* values might be due the MetMb% formation which canlead to the discoloration of the fresh meat (Krala, 2001).
Lipid oxidation
The oxidative stability of control group and experiment groups was evaluatedthroughout the storage by determining the TBARS as shown in Fig. 2. The TBARS value was continuously increasing in thecontrol group during the storage duration (p<0.05). Whereas, the TBARSvalues of Chinese cinnamon treated samples retarded during the storageintervals. Compared with experiment groups, the TBARS of control group wasnotably (p<0.05) higher during day 4 to 16. However, among alltreatments, the samples with 2.5% treatment exhibited the lower TBARSvalues after day 8 to 16 (p<0.05). The TBARS value for the control wassignificantly (p<0.05) higher at day 16 than that of Chinese cinnamontreated samples. The results suggested that Chinese cinnamon powder waseffective against the TBARS formation in the ground lamb during storage at4°C.
Fig. 2.
Effect of different concentrations of Chinese cinnamon powder on theTBARS values of vacuumed ground lamb meat stored at 4°C.
Different markers shows mean values while the bars indicated standarddeviations at each sampling point (n=3). A–DMean with different letters indicated changes between treatments differsignificantly (p<0.05). a–d Mean with differentletters showed changes during storage differ significantly(p<0.05). TBARS, thiobarbituric acid reactive substances.
The cinnamon contained active compounds, which can lead to antioxidant andantibacterial actions in meat (Madsen andBertelsen, 1995). Previous studies reported that the rapid increasein oxidation of control samples in the rainbow trout was due to non-availabilityof anti-oxidants (Shadman et al., 2017).The findings of present study agreed with the results of Shaltout et al. (2017), who also observed that the additionof cinnamon oil in beef delayed lipid oxidation during storage. Therefore, thereduction of TBARS in Chinese cinnamon treated samples may be caused by thepresence of antioxidant compounds like cinnamaldehyde, eugenol and cinnamic acid(Dudonne et al., 2009). Theseantioxidants compounds may be useful against free radical damage (Dragland et al., 2003).
Microbiological analysis
The effect of Chinese cinnamon powder on the TVC of ground lamb meat stored at4°C for 16 days was presented in Fig.3. The TVC of the control group increased immediately and rapidlythan other samples treated by Chinese cinnamon powder. At initial day, there wasno significant difference between the samples of the control and other treatedsamples. The TVC in control significantly increased (p<0.05) with theincrement of time points at day 4 and crossed the limit of 7 log CFU/g at 16days, while the treated samples retarded the growth of TVC. During the wholeperiod of storage, the samples with 1.5% treatment showed significantly(p<0.05) lower TVC values at days 8 and 16, and the lowest enumerationwas counted at day 8. From the obtained results, it was proved that TVC in theground lamb meat may be inhibited by the addition of Chinese cinnamon powder(p<0.05).
Fig. 3.
Effect of different concentrations of Chinese cinnamon powder on theTVC values of vacuumed ground lamb meat stored at 4°C.
Different markers shows mean values while the bars indicated standarddeviations at each sampling point (n=3). A–DMean with different letters indicated changes between treatments differsignificantly (p<0.05). a–d Mean with differentletters showed changes during storage differ significantly(p<0.05). TVC, total viable counts.
It has been reported that cinnamon inhibited microbes by several ways, such asrupturing of cell wall by the action antioxidant compounds, disordering thecytoplasmic membrane, cellular components disturbance by leakage, changed fattyacid and phospholipid constituents, affecting the DNA and RNA formation anddestroying protein translocation (Bajpai et al.,2013). Comparable results were obtained by Gutierrez et al. (2008), where it was reported that theaddition of cinnamon oil was more effective in decreasing the microbial countsin the food ingredients. In another study, cinnamon bark has been proved as apotential source against all pathogenic and spoilage bacteria (Ghabraie et al., 2016). Shaltout et al. (2017) observed that theincorporation of cinnamon oil was more efficient in maintaining meat quality.The reduction in the TVC during storage might be due to the presence ofbioactive compounds present in the Chinese cinnamon powder.
Conclusions
In conclusion, Chinese cinnamon powder could maintain the quality of ground lamb meatby reducing the TVC and TBARS. The L* and a* values in the ground lamb meat can beaffected during the storage by adding the Chinese cinnamon powder with aconcentration higher than 0.5%. The results of the present study suggestedthat Chinese cinnamon powder at a level of 0.5% had the potential to maintainthe ground lamb meat quality during storage. So it could be proposed as a naturalalternative of synthetic additives to maintain the meat quality.
Acknowledgments
The authors thank the financial support from the China Agriculture Research System(CARS-38), National Agricultural Science and Technology Innovation Program, ModernAgricultural Talent Support Program-Outstanding Talents and Innovative Team ofa*gricultural Scientific Research (2016-2020) and National High-level Personnel ofSpecial Support Program in China.
Conflicts of Interest
The authors declare no potential conflict of interest.
Author Contributions
Contributed by
Conceptualization: Zhang D. Methodology: Li X. Investigation: Hussain Z. Datacuration: Hussain Z, Li X, Ijaz M. Software: Ijaz M, Xiao X. Validation: Hou C,Zheng X, Ren C. Writing - original draft: Hussain Z. Writing-review &editing: Hussain Z, Li X, Ijaz M, Xiao X, Hou C, Zheng X, Ren C, Zhang D.
Ethics Approval
This article does not require IRB/IACUC approval because there are no human andanimal participants.
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