{"paper_id":"4de829a8-2d8a-47e1-8ddf-d091fe26ffa0","body_text":"Shelf life extension and meat quality enhancement of Labeo rohita during storage at refrigerated conditions using thyme oil and chitosan as natural preservatives | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Shelf life extension and meat quality enhancement of Labeo rohita during storage at refrigerated conditions using thyme oil and chitosan as natural preservatives Asma Nosheen, Abdul Majid Khan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-9124794/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 4 You are reading this latest preprint version Abstract The role of fishery in economy of a country with agricultural potential like Pakistan is very important. Fish meat has large market share as an important food source with high quality proteins, vitamins and minerals. Rohu ( Labeo rohita ) demand has increased due to consumer preference, market value and high yield in local environment. The huge economic loss due to spoilage at local and commercial level can be minimized by improving meat storage. In recent years the importance of natural organic preservatives has increased due to hazardous effects of synthetic preservatives. In this context essential oils in combination with chitosan have been proved a good edible meat preservative candidate. Keeping in view the importance of natural preservatives, the study was carried out to evaluate the preservation efficacy of thyme oil and chitosan separately and in combination on Rohu ( Labeo rohita ) meat preservation in refrigerated conditions (4℃). Following two-factor factorial completely randomized experimental design (CRD) was used. Thyme oil and chitosan as 0% & 1% in 2×2 interaction was applied on refrigerated fillets of Rohu meat. The treatment lasted 21 days and sampling was done on day 0, 7, 14 and 21 for analyses. Subsequently, results demonstrated that application of thyme oil (1%) in combination with chitosan (1%) extended shelf life of Rohu meat up to two weeks with maintained sensory acceptability. Hence, our study suggested the thyme oil and chitosan as promising edible organic preservative candidates for preservation of Rohu fillets in refrigerated conditions. Thyme oil and chitosan Natural preservatives Labeo rohita Shelf-life extension Preservation techniques Figures Figure 1 Figure 2 INTRODUCTION Fishery plays important role in the economy of a country as it brings home foreign exchange by export, provides employment and a source of livelihood for rural sector of society [1]. Both India and Bangladesh being major producer of the carp are increasing culturing of Rohu due to consumer demand, market value and high yield [2]. But despite the potential of industry, incompetence of the sector and lack of utilization of modern practices regarding post-harvest processing and preservation the producers in Pakistan fail to cover even the growing local supermarket demands, due to which the import of fish fillets increased from 5.4–17.7 US dollar [3]. Although the fishery industry in Pakistan could contribute 0.4% to the GDP in the year 2019-20 by earning 317.307 US dollars which is 2.7% (in quantity) higher as opposed to previous year [4], yet it is a drop in the ocean compared to the potential of the industry. Hindrance in this area is due to the perishable nature of the fish and poor processing and preservation techniques. Therefore, supply and storage of fish is challenging. This is an active research area with objectives to achieve preservation methods and techniques [5, 6]. An estimated amount of 11 billion US dollars on food losses was recorded in developing countries as a result of poor meat preservation [7]. Chemical preservatives include nitrite, nitrates, sulfates, sorbates benzoates formaldehyde, propionates etc. [8]. These preservatives though control the rancidity and extend shelf life effectively, but hazardous effects associated with these chemical preservatives range from minor skin rashes and allergies to the serious risk of carcinogenicity [9]. In the recent years, consumer awareness and safety concerns have extensively established market trends to search for safer natural preservatives as an alternate option [10]. Different natural substances having antimicrobial antioxidant properties are now being used as preservatives [11]. Thyme a member of family Lamiaceae is a well-known component of ethnomedicine [12]. Use of thyme as a spice and natural therapeutic agent has been practiced for generations in different parts of the world [13]. Along with many other natural spices oils and extracts of herbs, thyme oil has been proved to show promising results in preservation of food [14]. Owing to the active compounds most importantly phenolic compounds such as thymol, carvacrol, cymol and terpenine, thyme essential oil presents its strong antimicrobial, antioxidant and anti-inflammatory properties [15–17]. The antioxidant potential of essential oils predominantly relies on their chemical constituents and intricately linked to the presence of phenolic compounds which exhibit significant redox activity and are pivotal to neutralizing free radicals and decomposing peroxide which are reactive oxygen species that can cause cellular damage through oxidative stress [18]. The phenolic compounds facilitate effective neutralization of these reactive species through hydrogen atom transfer. This mechanism bears significant importance in the meat industry offering strategy for preservation and prevention from oxidative spoilage [19]. Extension in shelf life of meat of different origin including fish can not only maximize but also facilitate their marketing to distant areas and add to the revenue. Food contamination is regarded as a significant threat to consumer health and various bacterial strains can lead to the onset of food-borne illnesses [20, 21]. Antimicrobial property of essential oil arising from multiple mechanisms operating concurrently depending upon the chemical constituents they contain can keep the microbial growth at bay and hence prolong the shelf life with safety [22]. The present study was conducted to evaluate thyme essential oil and chitosan as alternative of chemical preservatives to extend shelf life of rohu fish meat. The results demonstrated that thyme oil in combination with chitosan coating may be used as natural preservative to extend shelf life up to two weeks in refrigerated conditions (4°C). MATERIALS AND METHODS Fish samples preparation and experimental design The Rohu fish (1 ± 0.5 kg average weight) was purchased alive from Head Qadirabad commercial fish market (Latitude, 32° 17' 52.51\"N, Longitude 73° 30' 0.71\"E) (Fig. 1 ) and then was beheaded, gutted and cleaned. The analysis was carried out immediately after harvesting. Four treatment lots (100 g) were assigned and two-factor factorial CRD experimental design was followed [23]. The meat samples were treated with chitosan and thyme oil having 0% & 1% concentration in interaction. The above mentioned concentration (s) was selected on the basis of literature mentioning sensory acceptability and consumer preference [24]. After treatment samples were packed in zip-locked polyethylene bags applying vacuum, labeled and stored at 4°C. Afterward, analyses were done with 7 days interval at day 0, 7, 14 and 21 of storage. Proximate composition analysis The proximate composition was evaluated by the determination of crude fat, moisture and protein by following standard methods of AOAC [25]. Briefly, muscle samples were homogenized well with mortar and pestle. Crude fat and protein were determined by using Soxhlet and Kjeldahl apparatus [26], respectively. The moisture contents were determined by oven drying the samples for 12 hrs at 105°C. Water-holding capacity (WHC) Muscles of Labeo rohita (5 g), minced in mortar and pestle and salt-induced method was used for WHC determination [27]. Each sample was centrifuged at 10,000 ×g for 15 min at room temperature and liquid was drained by inverting tubes for 30 min. The percentage weight gain of pellet was expressed as WHC. Protein extractability After homogenization of minced muscle (2.5 g) with 25 ml of Tris–HCl (0.03 M) the homogenate was centrifuged at 7,000 ×g for 15 min at 4°C. After separation of supernatant, remaining residues was again processed as mentioned above. The water extractable proteins (WEP) were determined in total 15 ml supernatant recovered. The remaining residual muscle material was used for salt extractable proteins (SEP) by homogenizing with 25 ml of Tris–HCl (0.03 M) and KCl (0.6 M) at pH 7 [23] with above mentioned method and protein concentration was determined by using Biuret method [28]. Total volatile basic nitrogen (TVBN) To determine TVBN, micro diffusion method was used by homogenizing fish sample (2 g) with Magnesium oxide (MgO) followed by distillation [29]. After collecting distillate in a flask containing mixed indicator (methylene blue + methyl red) and 3% of boric acid (H 3 BO 3 ), was incubated for color change (purple to green). Collection was continued for 2 times after colour change. Afterward distillate titration against 0.05 M of sulfuric acid was carried out and the consumed volume of sulfuric acid showed TVB-N content (mg/100g). The required chemicals/reagents and their compositions are as follow; Thiobarbituric acid reactive substances (TBARS) The measurement of TBARS was done by homogenization of sample in 3 ml Tris-maleate (80 mM, pH 7.4) and KCl (0.15 M) [30]. Briefly, lipid peroxidation was done by adding ascorbic acid to the sample and incubated at 37°C for 30 min. HCl (0.7 M) and Thiobarbituric acid (TBA) measuring 5 ml each were added followed by mixing with sample and boiled for 25 min. After cooling 5 ml of trichloroacetate (TCA) was further added and tubes were centrifuged at 495 xg for 5 minutes. Absorbance was measured at 530 nm using spectrophotometer and TBARS contents were estimated in terms of µg malondialdehyde (MDA) equivalents/mg tissue using standard solution of MDA. The chemicals/reagents and their compositions are as under; Total viable count (TVC) The TVC was carried out through spread plate method. Serial sample (0.1ml) dilution was spread on agar plate and subsequently colony forming units (CFU) were counted after overnight aerobic incubation at 37°C. The counted CFU/g was converted to Log 10 CFU/g [31]. Sensory assessment Highly qualified seven-member panel was requested to evaluate blind coded fish samples by using Quality Index Method (QIM) scheme through 5-point Hedonic scale. The assessment was based on considering color (1 extreme discoloration; 5 no discoloration), texture (1 very tender; 5 firm), odor (1 off odor/undesirable, 5 highly desirable), and general acceptability (1 unacceptable; 5 highly acceptable) [32]. The sensory score below three was considered as rejection of the sample in terms of shelf life criteria. Statistical analysis The CoStat computer package (Version 6.303, PMB320 Monterey, CA, 93940 USA) was used to analyze the observed data statistically. Two-way analysis of variance (ANOVA) was applied to determine the impact of treatments and shelf life assessment for meat samples. The p < 0.01 value was considered as significant for all analyses. RESULTS Analysis of the proximate composition The proximate composition analysis revealed significant (P < 0.01) linear decline in crude protein contents over the time of storage. Moreover, significant effect of treatment and time x treatment (P < 0.01) was observed. At the end of storage, the thyme oil and chitosan combination (18.09%) exhibited approximately 0.77% increased crude proteins contents compared to control (17.95%) (Table 1 ). Similarly, crude lipid contents decreased significantly in all samples during storage, with significant effect of treatment (P < 0.01) and time but not in case of time × treatment interaction (P = 0.07). Among the treated groups, the thyme oil × chitosan combination exhibited the highest crude lipid retention (4.42%) and, on day 21, contained approximately 1.8% more crude lipid content compared to the control (4.34%) (Table 1 ). Significant decrease in moisture content revealed in all samples during storage with significant effect of treatment and time × treatment interaction (P < 0.01). The treated groups exhibited statistically similar moisture content up to day 14; however, thereafter, the thyme oil x chitosan treatment showed superior retention of moisture content (74.93%), representing approximately 0.16% higher moisture retention compared to the control (74.81) (Table 1 ). Table 1 Effects of thyme oil and chitosan on proximate composition during of refrigerated (4°C) Rohu meat preservation Parameters Treatments Days 0 7 14 21 Crude Protein (%) Control 18.1 Aa 18.08 Bc 18.04 Bb 17.95 Cc Thyme Oil 18.1 Aa 18.10 Bbc 18.08 Ba 18.04 Cb Chitosan 18.15 Aa 18.12 Bab 18.10 Ba 18.07 Cab Thyme Oil x Chitosan 18.15 Aa 18.13 ABa 18.11 BCa 18.09 Ca SE 0.01 0.006 0.006 0.008 Crude Lipids (%) Control 4.54 Aa 4.47 Ba 4.41 Cb 4.34 Dc Thyme Oil 4.55 Aa 4.49 Ba 4.44 Ca 4.38 Db Chitosan 4.55 Aa 4.5 Ba 4.46 Ca 4.41 Dab Thyme Oil x Chitosan 4.55 Aa 4.51 Ba 4.47 Ca 4.42 Da SE 0.01 0.006 0.006 0.008 Moisture (%) Control 75.13 Aa 75.0 Bb 74.93 Cc 74.81 Dd Thyme Oil 75.13 Aa 75.1 Ba 74.99 Cb 74.84 Dc Chitosan 75.14 Aa 75.1 Ba 75.04 Ca 74.88 Db Thyme Oil x Chitosan 75.14 Aa 75.1 Aa 75.06 Ba 74.93 Ca SE 0.01 0.006 0.006 0.008 ANOVA (p-values) Treatments Time Time x Treatments Crude Protein (%) < 0.01 < 0.01 < 0.01 Crude Lipids (%) < 0.01 < 0.01 0.07 Moisture (%) < 0.01 < 0.01 < 0.01 Proteins extractability As a general trend, significant (P < 0.01) increase in WEP contents was observed over time in all samples with significant (P < 0.01) effect of treatments. However, the increase in WEP content was slower in treatment groups as compared to control (5.865 g/100 g wet weight). Interestingly, all treated groups showed same effect on WEP values during the course of preservation (5.78 g/100 g wet weight) with approximately 1.5% reduced protein solubility compared to control on day 21 of the storage (Table 4.16). On the other hand, significant (P < 0.01) decline in the extraction of salt soluble protein (SEP) in control and the treatment groups was observed over time from day zero to day 21. Contrastingly, decrease over time in SEP was observed in the treatment groups in following order; thyme oil > chitosan > combination of thyme oil and chitosan. Hence thyme oil and chitosan combination significantly restricted the change in SEP contents consistently over the time of storage (Table 2 ). On day 21 in thyme oil × chitosan- treated samples SEP contents (13.48 g/100 g wet weight) was approximately 1.9% higher than control (13.235 g/100 g wet weight). Table 2 Effects of thyme oil and chitosan on protein extractability of refrigerated (4°C) Rohu meat preservation Parameters Treatments Days 0 7 14 21 Salt Extractable Proteins (g/100 g wet weight) Control 13.685 Aa 13.54 Bb 13.365 Cc 13.235 Dc Thyme Oil 13.675 Aa 13.6 Ba 13.47 Cb 13.33 Db Chitosan 13.68 Aa 13.625 Ba 13.515 Ca 13.35 Db Thyme Oil x Chitosan 13.675 Aa 13.635 Aa 13.55 Ba 13.48 Ca SE 0.01 0.006 0.006 0.008 Water Extractable Proteins (g/100 g wet weight) Control 5.68 Da 5.73 Ca 5.785 Ba 5.865 Aa Thyme Oil 5.68 Ca 5.71 Cab 5.755 Bab 5.8 Ab Chitosan 5.675 Ca 5.705 Cab 5.75 Bb 5.81 Ab Thyme Oil x Chitosan 5.675 Ca 5.695 Cb 5.73 Bb 5.78 Ab SE 0.01 0.006 0.006 0.008 ANOVA (p-values) Treatments Time Time x Treatments Salt Extractable Proteins (g/100 g wet weight) < 0.01 < 0.01 < 0.01 Water Extractable Proteins (g/100 g wet weight) < 0.01 < 0.01 0.01 Water holding capacity The water holding capacity (WHC) of all treatments was significantly affected by time, treatment and their interaction (P < 0.01) during the storage period (Table 3 ). A greater decline in WHC was observed in the control samples () compared to the treated groups throughout storage. Among all treated groups, the thyme oil × chitosan combination (6.68%) showed the best performance, retaining approximately 2.7% higher water contents than the control (6.5%) on day 21 (Table 3 ). Table 3 Effects of thyme oil and chitosan on water holding capacity of refrigerated (4°C) Rohu meat preservation Parameters Treatments Days 0 7 14 21 Water Holding Capacity (%) Control 6.86 Aa 6.73 Bc 6.64 Cc 6.5 Dc Thyme Oil 6.86 Aa 6.77 Bb 6.71 Cb 6.65 Dab Chitosan 6.86 Aa 6.75 Bbc 6.72 Cb 6.63 Db Thyme Oil x Chitosan 6.86 Aa 6.80 Ba 6.76 Ca 6.68 Da SE 0.01 0.006 0.006 0.008 ANOVA (p-values) Treatments Time Time x Treatments < 0.01 < 0.01 < 0.01 Thiobarbituric acid reactive substances (TBARS) analysis The TABARS analysis showed a significant effect of time, treatment and their interaction on TBARS values during the period of storage (P < 0.01) (Table 4 ). The control samples exhibited high TBARS values during the course of storage till day 21 (2.56 mg MDA/kg) than treated groups. Like other parameters, in case of TBARS analysis thyme oil in combination with chitosan showed the best performance decreasing approximately 19.5% TBARS value (2.06 mg MDA/kg) than control on day 21 (Table 4 ). Table 4 Effects of thyme oil and chitosan on TBARS values of refrigerated (4°C) Rohu meat preservation Parameters Treatments Days 0 7 14 21 TBARS (mg melondialdehyde/kg meat) Control 1.24 Da 1.57 Ca 1.85 Ba 2.56 Aa Thyme Oil 1.23 Da 1.48 Cb 1.71 Bb 2.24 Ab Chitosan 1.23 Da 1.42 Cc 1.64 Bc 2.14 Ac Thyme Oil x Chitosan 1.25 Da 1.36 Cd 1.57 Bd 2.06 Ad SE 0.01 0.006 0.006 0.008 ANOVA (p-values ) Treatments Time Time x Treatments < 0.01 < 0.01 < 0.01 Peroxide value (POV) The POV of the samples was significantly affected by time, treatment and their interaction during the storage period from day zero to day 21 (P < 0.01) (Table 5 ). The control group exhibited higher POV values than all treated groups throughout the storage period. While, among all treated groups, thyme oil × chitosan combination showed the least POV value (16.12 mmol of O2/kg fats) on day 21 and decreased POV approximately 9.8% compared to control (17.87 mmol of O2/kg fats) (Table 5 ). Table 5 Effects of thyme oil and chitosan on POV of refrigerated (4°C) Rohu meat preservation Parameters Treatments Days 0 7 14 21 Peroxide Value (mmol of O2/kg fats) Control 11.170 Da 13.460 Ca 15.575 Ba 17.870 Aa Thyme Oil 11.155 Da 12.830 Cb 14.270 Bb 16.595 Ab Chitosan 11.160 Da 12.665 Cc 14.175 Bc 16.370 Ac Thyme Oil x Chitosan 11.145 Da 12.245 Cd 14.055 Bd 16.12 Ad SE 0.01 0.006 0.006 0.008 ANOVA (p-values) Treatments Time Time x Treatments < 0.01 < 0.01 < 0.01 Sensory attributes analysis All the sensory attributes evaluated in this study were significantly affect by time, treatment and their combination throughout the storage period (P < 0.01) (Table 6 ). Sensory score progressively declined in all samples, however, treated groups consistently exhibited higher values than the control from day zero to day 21. While among treated groups, the combination of thyme oil and chitosan showed best performance throughout the storage period, maintaining significantly firmness (33.6%), texture (14.5%), odor (9%) and colour (9.5%) values compared to control on day 21 (Table 6 ). Table 6 Effects of thyme oil and chitosan on sensory attributes of refrigerated (4°C) Rohu meat preservation Parameters Treatments Days 0 7 14 21 Firmness Control 9.80 Aa 8.66 Bc 6.77 Cd 5.65 Dd Thyme Oil 9.79 Aa 8.75 Bc 7.14 Cc 6.37 Dc Chitosan 9.90 Aa 8.84 Bb 7.25 Cb 6.63 Db Thyme Oil x Chitosan 9.80 Aa 8.94 Ba 7.57 Ca 6.84 Da SE 0.01 0.006 0.006 0.008 Texture Control 9.80 Aa 8.17 Bd 7.87 Cd 6.74 Dd Thyme Oil 9.80 Aa 8.54 Bc 8.26 Cc 7.29 Dc Chitosan 9.84 Aa 8.68 Bb 8.44 Cb 7.45 Db Thyme Oil x Chitosan 9.83 Aa 8.84 Ba 8.65 Ca 7.72 Da SE 0.01 0.006 0.006 0.008 Odor Control 9.93 Aa 8.60 Bd 7.36 Cd 6.65 Dd Thyme Oil 9.92 Aa 8.84 Bc 7.40 Cc 6.94 Dc Chitosan 9.92 Aa 8.92 Bb 7.76 Cb 7.15 Db Thyme Oil x Chitosan 9.93 Aa 9.12 Ba 7.92 Ca 7.27 Da SE 0.01 0.006 0.006 0.008 Colour Control 9.92 Aa 8.57 Bd 7.46 Cc 6.47 Dd Thyme Oil 9.92 Aa 8.70 Bc 7.85 Cb 6.76 Dc Chitosan 9.94 Aa 8.84 Bb 7.92 Cb 6.83 Db Thyme Oil x Chitosan 9.90 Aa 9.21 Ba 8.24 Ca 7.09 Da SE 0.01 0.006 0.006 0.008 ANOVA (p-values) Treatments Time Time x Treatments Firmness < 0.01 < 0.01 < 0.01 Texture < 0.01 < 0.01 < 0.01 Odor < 0.01 < 0.01 < 0.01 Colour < 0.01 < 0.01 < 0.01 Microbiological analysis The increase in number of colony forming units per gram (CFU/g) was observed during the preservation from day 0 to day 21, in all treated groups, but this increase was comparatively low in treatment groups as compared to control group (2–8.43 log CFU/g). The range of TVC values for thyme oil and chitosan treatment groups, was observed 2 to 4.2 log CFU/g from day 0 to 14th, while thyme oil and chitosan combination showed 2 to 3.9 log CFU/g range of TVC values. Further, it increased up to 6.2 log CFU/g at day 21st in independent treatment of thyme oil and chitosan but the combination of thyme oil and chitosan showed 5.9 log CFU/g TVC value. Hence, least TVC profile was found in thyme oil and chitosan combination which suggested it effectiveness against microbial activities in refrigerated Rohu meat (Fig. 2 ). DISCUSSION The present study revealed a linear decline in crude protein content in control group from day zero to day 21 (Table 1 ). The treatment groups on the other hand showed less decrease over time compared to control which clearly displays that chitosan and thyme essential oil both have a potential role in preservation of crude protein over time in refrigerated storage. This outcome is consistent with previous studies suggesting that natural preservatives including chitosan and essential oils minimize protein degradation over time by inhibiting oxidative stress and microbial damage [33, 34]. Chitosan is known for its film forming abilities to reduce the breakdown of protein on one hand and its barrier effect to seal oxygen provides an unfavorable environment for the microbes to flourish on the other hand [35]. Therefore, it may be predicted that these properties of chitosan might have better preserved the protein contents. Furthermore, thyme essential oil known for its antioxidant and antimicrobial properties also played its part to combat oxidative stress and microbial damage by acting against proteolytic enzymes that become active during microbial growth [36]. Among the treatment groups, chitosan and thyme oil alone in treatment performed same. However, the combination group outperformed in the retention of crude protein. This result can be demonstrated in terms of synergistic effect of both the treatment components because the semi permeable barrier provided by chitosan might have helped thyme essential oil to stay longer on the fillets and perform antioxidant and antimicrobial activities [37, 34]. On the contrary, thyme essential oil would have shown the reduced effectiveness alone due to volatile nature of its components which on other hand stayed longer in the presence of coating effect of chitosan. Shahbazi, [34] and Ojagh et al. [33] also reported the outperformance of natural antioxidants coated with biopolymers to avoid the spoilage of deteriorating and unstable foods during storage. Crude lipids percentage analysis revealed that lipid oxidation increased as the storage period increased (Table 1 ). This finding is in line with observation of [38] who suggested that oxidative stress over lipids increases with oxygen interaction which could define why control group showed a rapid and steady decline in lipid percentage over time as compared to treatment groups. Thyme essential oil due to its high phenolic compounds content leads to the formation of phenoxyl radical instead of peroxyl radical and hence terminate the chain reaction of lipid oxidation by breaking O-O bond [39, 38]. Present study aligns with these findings as thyme oil persistently depicted significantly better results as compared to control up until day 14 of experiment, later on chitosan and combination group took the lead. This difference could be explained on the basis of volatile phenolic compounds which could have stayed longer if coated with some barrier as in combination group where chitosan film appears to enhance the effectiveness of thyme oil longer than thyme oil alone. Findings are consistent with the research reported by Ojagh et al. [33], Shahbazi [34], and Muñoz-Tebar et al. [40]. With increasing storage time, the moisture contents gradually decreased in all the treatment groups as well as control (Table 1 ). But as the results indicate that treatment groups retained moisture level significantly better than control. These outcomes suggest that chitosan and thyme essential oil are effective in retaining moisture and hence contribute to the freshness over a longer period of time. Under storage conditions the protective effect of thyme oil and chitosan against moisture levels was also observed by Ojagh et al. [33] and Kanatt et al. [37]. Thyme oil performed equally well early on but later as the storage progressed the combination of both remarkably showed superior results. It is highly conceivable from the above results that the synergistic working of chitosan coating affect to overcome water loss and thyme oil’s antimicrobial effect worked hand in hand to stabilize fish fillets for longer storage by controlling microbial growth. Otherwise, proteolytic and lipolytic enzymes would cause denaturation of proteins and fats and increase moisture loss [41]. In addition, microbial by product of metabolism further augment moisture loss by destruction of cell membrane [42]. An increase in water extractable proteins over time was observed in all the treatment groups as well as control (Table 2 ). However, the treatment groups showed less increase consistent with the findings of Nawaz et al. [23]. This increase in water extractable protein overtime can be attributed to protein damage and formation of smaller peptides which increase solubility in water. This degradation of protein might be the result of enzymatic proteolysis of muscle proteins into smaller and more soluble peptides and amino acids, hence increasing water extractable proteins during preservation [43]. Furthermore, oxidation of proteins also expedites denaturation by unfolding the three dimensional structure and increasing the amount of water extractable proteins [33, 44, 45]. Antioxidant properties of thyme oil and film barrier provided by chitosan did alleviate denaturation to some degrees which was more pronounced in the combination group due to the dual effect working together. During storage protein denaturation due to oxidative stress results in breakdown of muscle protein and hence changes the three dimensional folding which leads to the changes in protein solubility [46]. Additionally, protein unfolding causes cross linking of some amino acids forming insoluble complexes further reducing the extractability of salt soluble proteins [47, 48]. The overall decline in the extraction of salt soluble proteins in control and that of treatment groups overtime can be attributed to the above mentioned phenomenon going on under storage condition (Table 2 ). Contrastingly, lesser decrease overtime in salt extractable proteins was observed in treatment group thyme oil to chitosan and combination groups consistently maintain protein integrity which result in exhibiting higher proportion of salt extractable proteins. Tironi et al. [49] also reported higher proportion of salt extractable proteins from rosemary essential oil in sea Salmon. The antioxidant potential of thyme essential oil and antimicrobial activity of chitosan seems to operate here to alleviate the cross linking effect of proteins by maintaining their structure integrity [46]. This protection was even enhanced in the combination and coating allowed the antioxidant properties of thyme oil worked longer. One of the freshness indicator of fish under storage conditions is its water holding capacity. In our study, control group showed a significant decline in water holding capacity over time as compared to treatment groups which can be attributed to the oxidative and microbial damage occurring during storage (Table 3 ). However, treatment groups retained moisture significantly better than control. Similar results were reported by Nawaz et al. [23], where chitosan coated samples showed better results than control to maintain water holding capacity. Thyme oil and chitosan alone in treatment performed in similar fashion in maintaining water holding capacity. But the combination took the lead which emphasizes the importance of antioxidant and antimicrobial properties of thyme oil and chitosan [40, 50]. The oxidation of lipids and proteins would otherwise effect negatively to retain moisture. Similarly, microbial load also affects the structural integrity of protein, further contributing to loss of water retained in muscle fibers. Therefore, it is conceivable that a combination of both compounds would have been preventing water loss by reducing structural denaturation and combating oxidative stress by combined efforts working together of chitosan coating and thyme essential oil [41]. The effectiveness of thyme oil further enhanced by chitosan coating can also be inferred from the work of Yu et al. [44] who reported the microencapsulation enhanced thyme oils performance to maintain the quality of meat under storage. Among commonly used methods to estimate lipid oxidation is TABARS value especially in fish meat [51]. MDA is taken as a marker of oxidative damage. It is a product of advanced stage deterioration which is measured by TBARS essay [12, 52]. An increasing trend in TBARS values with advancing storage days was observed in control, whereas the treatment groups showed significantly lower values compared to control throughout the storage period (Table 4 ). However, among the treatment groups the combination group revealed least TBARS values in terms of MDA/ kg of fish at all the sampling steps during experiment. In agreement with our results Yu et al. [44] also reported lower TABARS values of meat in thyme oil treatment. Another study by Ozogul et al. [53] also presented the powerful role of thyme essential oil in combating lipid oxidation compared to untreated group. Similarly, chitosan coated trout’s fillets with lemon verbena essential oil also exhibited lower TBARS values [54]. The study of Nawaz et al. [23] was also in line with our finding where combination group consisting of chitosan and rosemary essential oil demonstrated least values. The lower TBARS values advocate the strong antioxidant properties of thyme oil and coating nature of chitosan which also indirectly protects oxidative damage by minimizing oxygen exposure [55]. In combination group chitosan besides being excellent bioactive coating agent also exhibit antibacterial and antioxidant properties through which it attacks free radicals and terminate oxidative chain reaction. This antioxidant activity presents chitosan as a radical scavenger, scavenging aldehyde radicals e.g. MDA produced during lipid oxidation. This mechanism reportedly involves residual amino groups present in chitosan that form complex with MDA [37]. The concomitant action of the afore mentioned properties of chitosan and strong antioxidant power of thyme essential oil owe to phenolic compounds like carvacrol, thymol, and p-cymene seems to keep oxidative stress at bay, hence showing least TABARS values in case of combination group. Reports indicate that phenolic compounds present in thyme essential oil not only restrain the free radicals initiation by metal ion chelation [56] but also act as hydrogen atom donor from their hydroxyl group to produce phenoxyl radicals which in turn reacts with other peroxyl radicals and hence terminate the oxidative chain reaction [39, 57]. These findings are strongly suggestive of the use of natural essential oils like thyme oil with bioactive coating materials like chitosan etc. for foods rich in unsaturated fats like fish which otherwise make them prone to lipid oxidation causing rancidity, off flavours, hence limiting shelf life. Where TBARS gives secondary oxidation product value, POV gives primary lipids oxidation values for food over storage period [58]. Present study indicated an increasing trend in peroxide value with the passage of time in all the treated groups and control as well. However, all the treatment groups showed significantly better performance than control group (Table 5 ). But when thyme oil and chitosan coexisted in combination they performed best among all treated groups. This trend of increase in POV over time, may be attributed to the presence of several oxidation promoters in meat for example pro-oxidants like pigments containing hem or non-hem iron such as hemoglobin and myoglobin [59]. Similarly, percentage of lipid and protein also contribute towards oxidation promotion [60]. Comparable results were observed by Nawaz et al. [23] when rosemary essential oil was combined with chitosan to prevent oxidation of lipids. Chitosan formulated lemon extract and essential oil on rainbow trout also resulted in lower POV of samples treated compared to control [54]. In accordance with our work, a low POV was observed in salmon fillets coated in bioactive coating and cinnamon essential oil group demonstrating a clear control on oxidative stress by the phenolic compounds of essential oil and barrier provided by bioactive coating [61]. Multiple mechanisms are engaged by the phenolic compounds present in essential oils to alleviate oxidative stress. These mechanisms include controlling the first phase of oxidation to propagative phase by radical scavenging chain terminating steps through metal chelation and ROS prohibition [62, 18, 50]. Consumer acceptance and preference can be judged effectively on the basis of sensory analysis of stored and preserved food especially perishable ones like fish. Texture is the key indicator of quality under storage conditions [63]. Assessment of moisture and tenderness has been widely used to predict the texture of fish shelf life extension. In this regard panel of experts revealed that texture decline occurred in control as well as all the treatment groups over the period of storage. But that textural decline in all treatment groups was significantly lower compare to control (Table 7). In terms of juiciness and tenderness, the highest score for texture was observed in combination of thyme essential oil and chitosan group, which may be attributed to hygroscopic and film-forming properties of chitosan and antioxidant properties of thyme oil. Similar findings were reported by Yu et al. [44] for textual stability of mutton patties using thyme essential oil and microencapsulation. Odor assessment by the panel of experts revealed the same pattern as that observed in case of texture. Control group got least score at the end of the storage period that is 21st day whereas thyme oil and chitosan combination group got the highest acceptability score (Table 7). In consistence with our results Nawaz et al. [23] reported least acceptability of control and better score for rosemary essential oil and chitosan coated mori samples. Similarly, Li et al. [64] and Yu et al. [44] also reported similar patterns of results using microencapsulated clove and thyme essential oil, respectively. Microbial control by thyme oils is well established, antimicrobial nature and additive control in a synergistic manner by chitosan was effectively performed which resulted in odor inhibition and increased acceptability of samples. Furthermore, oxidative damage which also contributes to off flavours as well as off odor, was also reduced by the combination group as depicted by lower values of TABARS and TVBN [40, 44]. Colour changes occurred in all samples including control and treatment groups with significantly better colour preservation in treatment groups compare to control and highest score being obtained by combination group (Table 7). Similar results were obtained by Yu et al. [44] where higher colour preservation score was observed in microencapsulated thyme oil treated mutton parties compared to other group and control. Gum Arabic in Yu et al. [44] case seems to work similar as that of edible coating to minimize the volatility of phenolic compounds of thyme oil so that it may work synergistically against the deteriorative factors which cause colour change for example oxidative stress. Least score for firmness in case of control and highest score in case of combination group at the end of the storage period reveals that factors like prevention of autolysis, muscle protein degradation and lipid oxidation might have retained firmness level by the synergistic action of both the components. Bacteria are very important microbial agents involved in microbial degradation of meat and posing foodborne diseases based public health threat. The utilization of thyme oil as natural preservative to restrict microbial activities is well reported [65]. In our study, an increase in CFU/g was observed during the preservation from day 0 to day 21st in all treated groups. In case of control group it was high (2–8.43 log CFU/g) but the treated groups showed comparatively less increase and the range of TVC values for thyme oil, chitosan and combination of thyme oil and chitosan treatment groups, was observed 2 to 6.2 log CFU/g with least values (5.9 log CFU/g) for the combination of thyme oil and chitosan (Fig. 2 ). Marino et al. [66] demonstrated antimicrobial activity of thyme oil against nine Gram-negative and six Gram-positive bacterial strains among which Escherichia coli O157:H7 was found most sensitive strain. According to ICMSF, 1986 the upper acceptable limit of TVC is 6–7 log CFU/g. The current study showed that TVC values in all treated groups were in acceptable range (2–6.2 log CFU/g) from day 0 to day 21. Recommendations In this study best performance was showed by thyme oil in combination with chitosan. The estimated cost of treatment in combination was Rs. 128/Kg fish meat. This minimal cost addition would not only extend the shelf life of fish meat up to two weeks but also ensure food safety during storage and minimize loss due to spoilage. Declarations Funding declaration This study was not funded Ethics statement The fresh specimens of Labeo rohita were obtained from a licensed commercial fish market, and no live animals were subjected to experimental procedures. Author Contribution A.S. conducted all experimental work, data compilation and analysis, wrote the manuscript. A.M.K. designed the research work, supervised all research work and data analysis, edited and reviewed the manuscript Acknowledgement We acknowledge Dr. Syed Zakir Hussain Shah, Department of Zoology, University of Gujrat Pakistan, for his kind consideration of lab access during this research work. References WHO. The state of food security and nutrition in the world 2018. 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Cite Share Download PDF Status: Under Review Version 1 posted Reviewers invited by journal 04 May, 2026 Editor assigned by journal 17 Mar, 2026 Submission checks completed at journal 17 Mar, 2026 First submitted to journal 14 Mar, 2026 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-9124794\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":633832471,\"identity\":\"62fe8b62-eba4-4638-a9ac-4a4768e7e67a\",\"order_by\":0,\"name\":\"Asma Nosheen\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"University of the Punjab\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Asma\",\"middleName\":\"\",\"lastName\":\"Nosheen\",\"suffix\":\"\"},{\"id\":633832472,\"identity\":\"67dfd1c5-a80f-4a76-8edb-1c01916adf1a\",\"order_by\":1,\"name\":\"Abdul Majid Khan\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA5UlEQVRIie3SsQrCMBAG4JNAXK66KqJvIEQCuojPklJw8gGcRHBw0l3wITrpepLBRXB1rAidOji56tXiGjsK5l9yy5fcJQHw8fnNGMJ8qQrKl8ocQJQkQprSBKAgqMqR7pKulMGsUxf4uCEM2zEFVrlI/2TMYQtSNxfBXiOMdUy1yDgJGWMRMIxtsGsh2DAm1OQk5yQnDSaYMnmWIJf3KSonkgm9ibuxS8KzKMOzyEFvqyK9sbXIPf55Et2zKd9YdZEm2XTUXh9XtuEi/CjcRbGrLAqBbsAfhT6V/Gzyjfj4+Pj8WV6VFUehvHDuBwAAAABJRU5ErkJggg==\",\"orcid\":\"\",\"institution\":\"University of the Punjab\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Abdul\",\"middleName\":\"Majid\",\"lastName\":\"Khan\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2026-03-14 20:08:11\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-9124794/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-9124794/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":108986794,\"identity\":\"6e63cea1-f28a-4f85-9842-b5be5f88e574\",\"added_by\":\"auto\",\"created_at\":\"2026-05-11 12:56:05\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":182110,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eSite map of Head Qadir Abad fish market\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-9124794/v1/a48da0c809d74c25bd55a092.png\"},{\"id\":108986786,\"identity\":\"89e763c1-8ee4-423a-9afe-636b36e0457a\",\"added_by\":\"auto\",\"created_at\":\"2026-05-11 12:56:02\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":41187,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eEffect of thyme oil and chitosan on total viable count (TVC) of preserved Rohu meat. TO = thyme oil, Ch = chitosan\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-9124794/v1/b24b0a62860a9f4012da0f0e.png\"},{\"id\":108988024,\"identity\":\"fdba53d9-cf9e-4437-bb3d-cc6be226719a\",\"added_by\":\"auto\",\"created_at\":\"2026-05-11 13:03:08\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":768804,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-9124794/v1/909faae8-f52c-490f-b344-aaa5f36b0ee5.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Shelf life extension and meat quality enhancement of Labeo rohita during storage at refrigerated conditions using thyme oil and chitosan as natural preservatives\",\"fulltext\":[{\"header\":\"INTRODUCTION\",\"content\":\"\\u003cp\\u003eFishery plays important role in the economy of a country as it brings home foreign exchange by export, provides employment and a source of livelihood for rural sector of society [1]. Both India and Bangladesh being major producer of the carp are increasing culturing of Rohu due to consumer demand, market value and high yield [2]. But despite the potential of industry, incompetence of the sector and lack of utilization of modern practices regarding post-harvest processing and preservation the producers in Pakistan fail to cover even the growing local supermarket demands, due to which the import of fish fillets increased from 5.4\\u0026ndash;17.7 US dollar [3]. Although the fishery industry in Pakistan could contribute 0.4% to the GDP in the year 2019-20 by earning 317.307 US dollars which is 2.7% (in quantity) higher as opposed to previous year [4], yet it is a drop in the ocean compared to the potential of the industry. Hindrance in this area is due to the perishable nature of the fish and poor processing and preservation techniques. Therefore, supply and storage of fish is challenging. This is an active research area with objectives to achieve preservation methods and techniques [5, 6]. An estimated amount of 11\\u0026nbsp;billion US dollars on food losses was recorded in developing countries as a result of poor meat preservation [7].\\u003c/p\\u003e \\u003cp\\u003eChemical preservatives include nitrite, nitrates, sulfates, sorbates benzoates formaldehyde, propionates etc. [8]. These preservatives though control the rancidity and extend shelf life effectively, but hazardous effects associated with these chemical preservatives range from minor skin rashes and allergies to the serious risk of carcinogenicity [9]. In the recent years, consumer awareness and safety concerns have extensively established market trends to search for safer natural preservatives as an alternate option [10]. Different natural substances having antimicrobial antioxidant properties are now being used as preservatives [11].\\u003c/p\\u003e \\u003cp\\u003eThyme a member of family \\u003cem\\u003eLamiaceae\\u003c/em\\u003e is a well-known component of ethnomedicine [12]. Use of thyme as a spice and natural therapeutic agent has been practiced for generations in different parts of the world [13]. Along with many other natural spices oils and extracts of herbs, thyme oil has been proved to show promising results in preservation of food [14]. Owing to the active compounds most importantly phenolic compounds such as thymol, carvacrol, cymol and terpenine, thyme essential oil presents its strong antimicrobial, antioxidant and anti-inflammatory properties [15\\u0026ndash;17]. The antioxidant potential of essential oils predominantly relies on their chemical constituents and intricately linked to the presence of phenolic compounds which exhibit significant redox activity and are pivotal to neutralizing free radicals and decomposing peroxide which are reactive oxygen species that can cause cellular damage through oxidative stress [18]. The phenolic compounds facilitate effective neutralization of these reactive species through hydrogen atom transfer. This mechanism bears significant importance in the meat industry offering strategy for preservation and prevention from oxidative spoilage [19]. Extension in shelf life of meat of different origin including fish can not only maximize but also facilitate their marketing to distant areas and add to the revenue. Food contamination is regarded as a significant threat to consumer health and various bacterial strains can lead to the onset of food-borne illnesses [20, 21]. Antimicrobial property of essential oil arising from multiple mechanisms operating concurrently depending upon the chemical constituents they contain can keep the microbial growth at bay and hence prolong the shelf life with safety [22].\\u003c/p\\u003e \\u003cp\\u003eThe present study was conducted to evaluate thyme essential oil and chitosan as alternative of chemical preservatives to extend shelf life of rohu fish meat. The results demonstrated that thyme oil in combination with chitosan coating may be used as natural preservative to extend shelf life up to two weeks in refrigerated conditions (4\\u0026deg;C).\\u003c/p\\u003e\"},{\"header\":\"MATERIALS AND METHODS\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eFish samples preparation and experimental design\\u003c/h2\\u003e \\u003cp\\u003eThe Rohu fish (1\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.5 kg average weight) was purchased alive from Head Qadirabad commercial fish market (Latitude, 32\\u0026deg; 17' 52.51\\\"N, Longitude 73\\u0026deg; 30' 0.71\\\"E) (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e) and then was beheaded, gutted and cleaned. The analysis was carried out immediately after harvesting. Four treatment lots (100 g) were assigned and two-factor factorial CRD experimental design was followed [23]. The meat samples were treated with chitosan and thyme oil having 0% \\u0026amp; 1% concentration in interaction. The above mentioned concentration (s) was selected on the basis of literature mentioning sensory acceptability and consumer preference [24]. After treatment samples were packed in zip-locked polyethylene bags applying vacuum, labeled and stored at 4\\u0026deg;C. Afterward, analyses were done with 7 days interval at day 0, 7, 14 and 21 of storage.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e\\n\\u003ch3\\u003eProximate composition analysis\\u003c/h3\\u003e\\n\\u003cp\\u003eThe proximate composition was evaluated by the determination of crude fat, moisture and protein by following standard methods of AOAC [25]. Briefly, muscle samples were homogenized well with mortar and pestle. Crude fat and protein were determined by using Soxhlet and Kjeldahl apparatus [26], respectively. The moisture contents were determined by oven drying the samples for 12 hrs at 105\\u0026deg;C.\\u003c/p\\u003e\\n\\u003ch3\\u003eWater-holding capacity (WHC)\\u003c/h3\\u003e\\n\\u003cp\\u003eMuscles of \\u003cem\\u003eLabeo rohita\\u003c/em\\u003e (5 g), minced in mortar and pestle and salt-induced method was used for WHC determination [27]. Each sample was centrifuged at 10,000 \\u0026times;g for 15 min at room temperature and liquid was drained by inverting tubes for 30 min. The percentage weight gain of pellet was expressed as WHC.\\u003c/p\\u003e\\n\\u003ch3\\u003eProtein extractability\\u003c/h3\\u003e\\n\\u003cp\\u003eAfter homogenization of minced muscle (2.5 g) with 25 ml of Tris\\u0026ndash;HCl (0.03 M) the homogenate was centrifuged at 7,000 \\u0026times;g for 15 min at 4\\u0026deg;C. After separation of supernatant, remaining residues was again processed as mentioned above. The water extractable proteins (WEP) were determined in total 15 ml supernatant recovered. The remaining residual muscle material was used for salt extractable proteins (SEP) by homogenizing with 25 ml of Tris\\u0026ndash;HCl (0.03 M) and KCl (0.6 M) at pH 7 [23] with above mentioned method and protein concentration was determined by using Biuret method [28].\\u003c/p\\u003e\\n\\u003ch3\\u003eTotal volatile basic nitrogen (TVBN)\\u003c/h3\\u003e\\n\\u003cp\\u003eTo determine TVBN, micro diffusion method was used by homogenizing fish sample (2 g) with Magnesium oxide (MgO) followed by distillation [29]. After collecting distillate in a flask containing mixed indicator (methylene blue\\u0026thinsp;+\\u0026thinsp;methyl red) and 3% of boric acid (H\\u003csub\\u003e3\\u003c/sub\\u003eBO\\u003csub\\u003e3\\u003c/sub\\u003e), was incubated for color change (purple to green). Collection was continued for 2 times after colour change. Afterward distillate titration against 0.05 M of sulfuric acid was carried out and the consumed volume of sulfuric acid showed TVB-N content (mg/100g). The required chemicals/reagents and their compositions are as follow;\\u003c/p\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eThiobarbituric acid reactive substances (TBARS)\\u003c/h2\\u003e \\u003cp\\u003eThe measurement of TBARS was done by homogenization of sample in 3 ml Tris-maleate (80 mM, pH 7.4) and KCl (0.15 M) [30]. Briefly, lipid peroxidation was done by adding ascorbic acid to the sample and incubated at 37\\u0026deg;C for 30 min. HCl (0.7 M) and Thiobarbituric acid (TBA) measuring 5 ml each were added followed by mixing with sample and boiled for 25 min. After cooling 5 ml of trichloroacetate (TCA) was further added and tubes were centrifuged at 495 xg for 5 minutes. Absorbance was measured at 530 nm using spectrophotometer and TBARS contents were estimated in terms of \\u0026micro;g malondialdehyde (MDA) equivalents/mg tissue using standard solution of MDA. The chemicals/reagents and their compositions are as under;\\u003c/p\\u003e \\u003c/div\\u003e\\n\\u003ch3\\u003eTotal viable count (TVC)\\u003c/h3\\u003e\\n\\u003cp\\u003eThe TVC was carried out through spread plate method. Serial sample (0.1ml) dilution was spread on agar plate and subsequently colony forming units (CFU) were counted after overnight aerobic incubation at 37\\u0026deg;C. The counted CFU/g was converted to Log\\u003csub\\u003e10\\u003c/sub\\u003e CFU/g [31].\\u003c/p\\u003e\\n\\u003ch3\\u003eSensory assessment\\u003c/h3\\u003e\\n\\u003cp\\u003eHighly qualified seven-member panel was requested to evaluate blind coded fish samples by using Quality Index Method (QIM) scheme through 5-point Hedonic scale. The assessment was based on considering color (1 extreme discoloration; 5 no discoloration), texture (1 very tender; 5 firm), odor (1 off odor/undesirable, 5 highly desirable), and general acceptability (1 unacceptable; 5 highly acceptable) [32]. The sensory score below three was considered as rejection of the sample in terms of shelf life criteria.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eStatistical analysis\\u003c/h2\\u003e \\u003cp\\u003eThe CoStat computer package (Version 6.303, PMB320 Monterey, CA, 93940 USA) was used to analyze the observed data statistically. Two-way analysis of variance (ANOVA) was applied to determine the impact of treatments and shelf life assessment for meat samples. The p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01 value was considered as significant for all analyses.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"RESULTS\",\"content\":\"\\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eAnalysis of the proximate composition\\u003c/h2\\u003e \\u003cp\\u003eThe proximate composition analysis revealed significant (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) linear decline in crude protein contents over the time of storage. Moreover, significant effect of treatment and time x treatment (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) was observed. At the end of storage, the thyme oil and chitosan combination (18.09%) exhibited approximately 0.77% increased crude proteins contents compared to control (17.95%) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). Similarly, crude lipid contents decreased significantly in all samples during storage, with significant effect of treatment (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) and time but not in case of time \\u0026times; treatment interaction (P\\u0026thinsp;=\\u0026thinsp;0.07). Among the treated groups, the thyme oil \\u0026times; chitosan combination exhibited the highest crude lipid retention (4.42%) and, on day 21, contained approximately 1.8% more crude lipid content compared to the control (4.34%) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). Significant decrease in moisture content revealed in all samples during storage with significant effect of treatment and time \\u0026times; treatment interaction (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01). The treated groups exhibited statistically similar moisture content up to day 14; however, thereafter, the thyme oil x chitosan treatment showed superior retention of moisture content (74.93%), representing approximately 0.16% higher moisture retention compared to the control (74.81) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eEffects of thyme oil and chitosan on proximate composition during of refrigerated (4\\u0026deg;C) Rohu meat preservation\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"6\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eParameters\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"4\\\" nameend=\\\"c6\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eCrude Protein (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18.1 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e18.08 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e18.04 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e17.95 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18.1 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e18.10 Bbc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e18.08 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e18.04 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18.15 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e18.12 Bab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e18.10 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e18.07 Cab\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e18.15 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e18.13 ABa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e18.11 BCa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e18.09 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eCrude Lipids (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.54 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e4.47 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e4.41 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e4.34 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.55 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e4.49 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e4.44 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e4.38 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.55 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e4.5 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e4.46 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e4.41 Dab\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e4.55 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e4.51 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e4.47 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e4.42 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eMoisture (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75.13 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e75.0 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e74.93 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e74.81 Dd\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75.13 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e75.1 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e74.99 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e74.84 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75.14 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e75.1 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e75.04 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e74.88 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e75.14 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e75.1 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e75.06 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e74.93 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"6\\\" nameend=\\\"c6\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eANOVA (p-values)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eTime\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003eTime x Treatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eCrude Protein (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eCrude Lipids (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e0.07\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eMoisture (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eProteins extractability\\u003c/h2\\u003e \\u003cp\\u003eAs a general trend, significant (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) increase in WEP contents was observed over time in all samples with significant (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) effect of treatments. However, the increase in WEP content was slower in treatment groups as compared to control (5.865 g/100 g wet weight). Interestingly, all treated groups showed same effect on WEP values during the course of preservation (5.78 g/100 g wet weight) with approximately 1.5% reduced protein solubility compared to control on day 21 of the storage (Table\\u0026nbsp;4.16). On the other hand, significant (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) decline in the extraction of salt soluble protein (SEP) in control and the treatment groups was observed over time from day zero to day 21. Contrastingly, decrease over time in SEP was observed in the treatment groups in following order; thyme oil\\u0026thinsp;\\u0026gt;\\u0026thinsp;chitosan\\u0026thinsp;\\u0026gt;\\u0026thinsp;combination of thyme oil and chitosan. Hence thyme oil and chitosan combination significantly restricted the change in SEP contents consistently over the time of storage (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). On day 21 in thyme oil \\u0026times; chitosan- treated samples SEP contents (13.48 g/100 g wet weight) was approximately 1.9% higher than control (13.235 g/100 g wet weight).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eEffects of thyme oil and chitosan on protein extractability of refrigerated (4\\u0026deg;C) Rohu meat preservation\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"7\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eParameters\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c7\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eSalt Extractable Proteins (g/100 g wet weight)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13.685 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13.54 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e13.365 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e13.235 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13.675 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13.6 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e13.47 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e13.33 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13.68 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13.625 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e13.515 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e13.35 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e13.675 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13.635 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e13.55 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e13.48 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eWater Extractable Proteins (g/100 g wet weight)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.68 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5.73 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e5.785 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e5.865 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"1\\\" nameend=\\\"c7\\\" namest=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.68 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5.71 Cab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e5.755 Bab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e5.8 Ab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"1\\\" nameend=\\\"c7\\\" namest=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.675 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5.705 Cab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e5.75 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e5.81 Ab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"1\\\" nameend=\\\"c7\\\" namest=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e5.675 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e5.695 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e5.73 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e5.78 Ab\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"1\\\" nameend=\\\"c7\\\" namest=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"1\\\" nameend=\\\"c7\\\" namest=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"7\\\" nameend=\\\"c7\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eANOVA (p-values)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eTime\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003eTime x Treatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eSalt Extractable Proteins (g/100 g wet weight)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eWater Extractable Proteins (g/100 g wet weight)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eWater holding capacity\\u003c/h2\\u003e \\u003cp\\u003eThe water holding capacity (WHC) of all treatments was significantly affected by time, treatment and their interaction (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) during the storage period (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e). A greater decline in WHC was observed in the control samples () compared to the treated groups throughout storage. Among all treated groups, the thyme oil \\u0026times; chitosan combination (6.68%) showed the best performance, retaining approximately 2.7% higher water contents than the control (6.5%) on day 21 (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eEffects of thyme oil and chitosan on water holding capacity of refrigerated (4\\u0026deg;C) Rohu meat preservation\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"6\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eParameters\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"4\\\" nameend=\\\"c6\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eWater Holding Capacity (%)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.86 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.73 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e6.64 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.5 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.86 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.77 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e6.71 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.65 Dab\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.86 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.75 Bbc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e6.72 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.63 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e6.86 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e6.80 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e6.76 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e6.68 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"6\\\" nameend=\\\"c6\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eANOVA (p-values)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eTime\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003eTime x Treatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec16\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eThiobarbituric acid reactive substances (TBARS) analysis\\u003c/h2\\u003e \\u003cp\\u003eThe TABARS analysis showed a significant effect of time, treatment and their interaction on TBARS values during the period of storage (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e). The control samples exhibited high TBARS values during the course of storage till day 21 (2.56 mg MDA/kg) than treated groups. Like other parameters, in case of TBARS analysis thyme oil in combination with chitosan showed the best performance decreasing approximately 19.5% TBARS value (2.06 mg MDA/kg) than control on day 21 (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab4\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 4\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eEffects of thyme oil and chitosan on TBARS values of refrigerated (4\\u0026deg;C) Rohu meat preservation\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"6\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eParameters\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"4\\\" nameend=\\\"c6\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eTBARS (mg melondialdehyde/kg meat)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.24 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.57 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1.85 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2.56 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.23 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.48 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1.71 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2.24 Ab\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.23 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.42 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1.64 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2.14 Ac\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e1.25 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e1.36 Cd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e1.57 Bd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e2.06 Ad\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"6\\\" nameend=\\\"c6\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eANOVA (p-values\\u003c/b\\u003e)\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eTime\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003eTime x Treatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec17\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003ePeroxide value (POV)\\u003c/h2\\u003e \\u003cp\\u003eThe POV of the samples was significantly affected by time, treatment and their interaction during the storage period from day zero to day 21 (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e). The control group exhibited higher POV values than all treated groups throughout the storage period. While, among all treated groups, thyme oil \\u0026times; chitosan combination showed the least POV value (16.12 mmol of O2/kg fats) on day 21 and decreased POV approximately 9.8% compared to control (17.87 mmol of O2/kg fats) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab5\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 5\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eEffects of thyme oil and chitosan on POV of refrigerated (4\\u0026deg;C) Rohu meat preservation\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"6\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eParameters\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"4\\\" nameend=\\\"c6\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003ePeroxide Value (mmol of O2/kg fats)\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e11.170 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e13.460 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e15.575 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e17.870 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e11.155 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12.830 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14.270 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e16.595 Ab\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e11.160 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12.665 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14.175 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e16.370 Ac\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e11.145 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e12.245 Cd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e14.055 Bd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e16.12 Ad\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"6\\\" nameend=\\\"c6\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eANOVA (p-values)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eTime\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003eTime x Treatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec18\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eSensory attributes analysis\\u003c/h2\\u003e \\u003cp\\u003eAll the sensory attributes evaluated in this study were significantly affect by time, treatment and their combination throughout the storage period (P\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.01) (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e). Sensory score progressively declined in all samples, however, treated groups consistently exhibited higher values than the control from day zero to day 21. While among treated groups, the combination of thyme oil and chitosan showed best performance throughout the storage period, maintaining significantly firmness (33.6%), texture (14.5%), odor (9%) and colour (9.5%) values compared to control on day 21 (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab6\\\" class=\\\"InternalRef\\\"\\u003e6\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab6\\\" border=\\\"1\\\"\\u003e \\u003ccaption language=\\\"En\\\"\\u003e \\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 6\\u003c/div\\u003e \\u003cdiv class=\\\"CaptionContent\\\"\\u003e \\u003cp\\u003eEffects of thyme oil and chitosan on sensory attributes of refrigerated (4\\u0026deg;C) Rohu meat preservation\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/caption\\u003e \\u003ccolgroup cols=\\\"7\\\"\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e \\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e \\u003cthead\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eParameters\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c2\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"5\\\" nameend=\\\"c7\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eDays\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e7\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e14\\u003c/p\\u003e \\u003c/th\\u003e \\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e21\\u003c/p\\u003e \\u003c/th\\u003e \\u003c/tr\\u003e \\u003c/thead\\u003e \\u003ctbody\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eFirmness\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.80 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.66 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e6.77 Cd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e5.65 Dd\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.79 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.75 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e7.14 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e6.37 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.90 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.84 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e7.25 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e6.63 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.80 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.94 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e7.57 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e6.84 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c6\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eTexture\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.80 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.17 Bd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.87 Cd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e6.74 Dd\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.80 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.54 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8.26 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e7.29 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.84 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.68 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8.44 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e7.45 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.83 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.84 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8.65 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e7.72 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eOdor\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.93 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.60 Bd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.36 Cd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e6.65 Dd\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.92 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.84 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.40 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e6.94 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.92 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.92 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.76 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e7.15 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.93 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e9.12 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.92 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e7.27 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\" morerows=\\\"3\\\" rowspan=\\\"4\\\"\\u003e \\u003cp\\u003eColour\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eControl\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.92 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.57 Bd\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.46 Cc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e6.47 Dd\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.92 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.70 Bc\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.85 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e6.76 Dc\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eChitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.94 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e8.84 Bb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e7.92 Cb\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e6.83 Db\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eThyme Oil x Chitosan\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e9.90 Aa\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e9.21 Ba\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e8.24 Ca\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e7.09 Da\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eSE\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e \\u003cp\\u003e0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e \\u003cp\\u003e0.006\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c7\\\" namest=\\\"c6\\\"\\u003e \\u003cp\\u003e0.008\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"7\\\" nameend=\\\"c7\\\" namest=\\\"c1\\\"\\u003e \\u003cp\\u003e\\u003cb\\u003eANOVA (p-values)\\u003c/b\\u003e\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003eTreatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003eTime\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003eTime x Treatments\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eFirmness\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eTexture\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eOdor\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003ctr\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e \\u003cp\\u003eColour\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"2\\\" nameend=\\\"c4\\\" namest=\\\"c3\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003ctd align=\\\"left\\\" colspan=\\\"3\\\" nameend=\\\"c7\\\" namest=\\\"c5\\\"\\u003e \\u003cp\\u003e\\u0026lt;\\u0026thinsp;0.01\\u003c/p\\u003e \\u003c/td\\u003e \\u003c/tr\\u003e \\u003c/tbody\\u003e \\u003c/colgroup\\u003e \\u003c/table\\u003e\\u003c/div\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec19\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eMicrobiological analysis\\u003c/h2\\u003e \\u003cp\\u003eThe increase in number of colony forming units per gram (CFU/g) was observed during the preservation from day 0 to day 21, in all treated groups, but this increase was comparatively low in treatment groups as compared to control group (2\\u0026ndash;8.43 log CFU/g). The range of TVC values for thyme oil and chitosan treatment groups, was observed 2 to 4.2 log CFU/g from day 0 to 14th, while thyme oil and chitosan combination showed 2 to 3.9 log CFU/g range of TVC values. Further, it increased up to 6.2 log CFU/g at day 21st in independent treatment of thyme oil and chitosan but the combination of thyme oil and chitosan showed 5.9 log CFU/g TVC value. Hence, least TVC profile was found in thyme oil and chitosan combination which suggested it effectiveness against microbial activities in refrigerated Rohu meat (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"DISCUSSION\",\"content\":\"\\u003cp\\u003eThe present study revealed a linear decline in crude protein content in control group from day zero to day 21 (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). The treatment groups on the other hand showed less decrease over time compared to control which clearly displays that chitosan and thyme essential oil both have a potential role in preservation of crude protein over time in refrigerated storage. This outcome is consistent with previous studies suggesting that natural preservatives including chitosan and essential oils minimize protein degradation over time by inhibiting oxidative stress and microbial damage [33, 34]. Chitosan is known for its film forming abilities to reduce the breakdown of protein on one hand and its barrier effect to seal oxygen provides an unfavorable environment for the microbes to flourish on the other hand [35]. Therefore, it may be predicted that these properties of chitosan might have better preserved the protein contents. Furthermore, thyme essential oil known for its antioxidant and antimicrobial properties also played its part to combat oxidative stress and microbial damage by acting against proteolytic enzymes that become active during microbial growth [36]. Among the treatment groups, chitosan and thyme oil alone in treatment performed same. However, the combination group outperformed in the retention of crude protein. This result can be demonstrated in terms of synergistic effect of both the treatment components because the semi permeable barrier provided by chitosan might have helped thyme essential oil to stay longer on the fillets and perform antioxidant and antimicrobial activities [37, 34]. On the contrary, thyme essential oil would have shown the reduced effectiveness alone due to volatile nature of its components which on other hand stayed longer in the presence of coating effect of chitosan. Shahbazi, [34] and Ojagh et al. [33] also reported the outperformance of natural antioxidants coated with biopolymers to avoid the spoilage of deteriorating and unstable foods during storage.\\u003c/p\\u003e \\u003cp\\u003eCrude lipids percentage analysis revealed that lipid oxidation increased as the storage period increased (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). This finding is in line with observation of [38] who suggested that oxidative stress over lipids increases with oxygen interaction which could define why control group showed a rapid and steady decline in lipid percentage over time as compared to treatment groups. Thyme essential oil due to its high phenolic compounds content leads to the formation of phenoxyl radical instead of peroxyl radical and hence terminate the chain reaction of lipid oxidation by breaking O-O bond [39, 38]. Present study aligns with these findings as thyme oil persistently depicted significantly better results as compared to control up until day 14 of experiment, later on chitosan and combination group took the lead. This difference could be explained on the basis of volatile phenolic compounds which could have stayed longer if coated with some barrier as in combination group where chitosan film appears to enhance the effectiveness of thyme oil longer than thyme oil alone. Findings are consistent with the research reported by Ojagh et al. [33], Shahbazi [34], and Mu\\u0026ntilde;oz-Tebar et al. [40].\\u003c/p\\u003e \\u003cp\\u003eWith increasing storage time, the moisture contents gradually decreased in all the treatment groups as well as control (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e). But as the results indicate that treatment groups retained moisture level significantly better than control. These outcomes suggest that chitosan and thyme essential oil are effective in retaining moisture and hence contribute to the freshness over a longer period of time. Under storage conditions the protective effect of thyme oil and chitosan against moisture levels was also observed by Ojagh et al. [33] and Kanatt et al. [37]. Thyme oil performed equally well early on but later as the storage progressed the combination of both remarkably showed superior results. It is highly conceivable from the above results that the synergistic working of chitosan coating affect to overcome water loss and thyme oil\\u0026rsquo;s antimicrobial effect worked hand in hand to stabilize fish fillets for longer storage by controlling microbial growth. Otherwise, proteolytic and lipolytic enzymes would cause denaturation of proteins and fats and increase moisture loss [41]. In addition, microbial by product of metabolism further augment moisture loss by destruction of cell membrane [42].\\u003c/p\\u003e \\u003cp\\u003eAn increase in water extractable proteins over time was observed in all the treatment groups as well as control (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). However, the treatment groups showed less increase consistent with the findings of Nawaz et al. [23]. This increase in water extractable protein overtime can be attributed to protein damage and formation of smaller peptides which increase solubility in water. This degradation of protein might be the result of enzymatic proteolysis of muscle proteins into smaller and more soluble peptides and amino acids, hence increasing water extractable proteins during preservation [43]. Furthermore, oxidation of proteins also expedites denaturation by unfolding the three dimensional structure and increasing the amount of water extractable proteins [33, 44, 45]. Antioxidant properties of thyme oil and film barrier provided by chitosan did alleviate denaturation to some degrees which was more pronounced in the combination group due to the dual effect working together. During storage protein denaturation due to oxidative stress results in breakdown of muscle protein and hence changes the three dimensional folding which leads to the changes in protein solubility [46]. Additionally, protein unfolding causes cross linking of some amino acids forming insoluble complexes further reducing the extractability of salt soluble proteins [47, 48]. The overall decline in the extraction of salt soluble proteins in control and that of treatment groups overtime can be attributed to the above mentioned phenomenon going on under storage condition (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). Contrastingly, lesser decrease overtime in salt extractable proteins was observed in treatment group thyme oil to chitosan and combination groups consistently maintain protein integrity which result in exhibiting higher proportion of salt extractable proteins. Tironi et al. [49] also reported higher proportion of salt extractable proteins from rosemary essential oil in sea Salmon. The antioxidant potential of thyme essential oil and antimicrobial activity of chitosan seems to operate here to alleviate the cross linking effect of proteins by maintaining their structure integrity [46]. This protection was even enhanced in the combination and coating allowed the antioxidant properties of thyme oil worked longer.\\u003c/p\\u003e \\u003cp\\u003eOne of the freshness indicator of fish under storage conditions is its water holding capacity. In our study, control group showed a significant decline in water holding capacity over time as compared to treatment groups which can be attributed to the oxidative and microbial damage occurring during storage (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e). However, treatment groups retained moisture significantly better than control. Similar results were reported by Nawaz et al. [23], where chitosan coated samples showed better results than control to maintain water holding capacity. Thyme oil and chitosan alone in treatment performed in similar fashion in maintaining water holding capacity. But the combination took the lead which emphasizes the importance of antioxidant and antimicrobial properties of thyme oil and chitosan [40, 50]. The oxidation of lipids and proteins would otherwise effect negatively to retain moisture. Similarly, microbial load also affects the structural integrity of protein, further contributing to loss of water retained in muscle fibers. Therefore, it is conceivable that a combination of both compounds would have been preventing water loss by reducing structural denaturation and combating oxidative stress by combined efforts working together of chitosan coating and thyme essential oil [41]. The effectiveness of thyme oil further enhanced by chitosan coating can also be inferred from the work of Yu et al. [44] who reported the microencapsulation enhanced thyme oils performance to maintain the quality of meat under storage.\\u003c/p\\u003e \\u003cp\\u003eAmong commonly used methods to estimate lipid oxidation is TABARS value especially in fish meat [51]. MDA is taken as a marker of oxidative damage. It is a product of advanced stage deterioration which is measured by TBARS essay [12, 52]. An increasing trend in TBARS values with advancing storage days was observed in control, whereas the treatment groups showed significantly lower values compared to control throughout the storage period (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab4\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003e). However, among the treatment groups the combination group revealed least TBARS values in terms of MDA/ kg of fish at all the sampling steps during experiment. In agreement with our results Yu et al. [44] also reported lower TABARS values of meat in thyme oil treatment. Another study by Ozogul et al. [53] also presented the powerful role of thyme essential oil in combating lipid oxidation compared to untreated group. Similarly, chitosan coated trout\\u0026rsquo;s fillets with lemon verbena essential oil also exhibited lower TBARS values [54]. The study of Nawaz et al. [23] was also in line with our finding where combination group consisting of chitosan and rosemary essential oil demonstrated least values. The lower TBARS values advocate the strong antioxidant properties of thyme oil and coating nature of chitosan which also indirectly protects oxidative damage by minimizing oxygen exposure [55]. In combination group chitosan besides being excellent bioactive coating agent also exhibit antibacterial and antioxidant properties through which it attacks free radicals and terminate oxidative chain reaction. This antioxidant activity presents chitosan as a radical scavenger, scavenging aldehyde radicals e.g. MDA produced during lipid oxidation. This mechanism reportedly involves residual amino groups present in chitosan that form complex with MDA [37]. The concomitant action of the afore mentioned properties of chitosan and strong antioxidant power of thyme essential oil owe to phenolic compounds like carvacrol, thymol, and p-cymene seems to keep oxidative stress at bay, hence showing least TABARS values in case of combination group. Reports indicate that phenolic compounds present in thyme essential oil not only restrain the free radicals initiation by metal ion chelation [56] but also act as hydrogen atom donor from their hydroxyl group to produce phenoxyl radicals which in turn reacts with other peroxyl radicals and hence terminate the oxidative chain reaction [39, 57]. These findings are strongly suggestive of the use of natural essential oils like thyme oil with bioactive coating materials like chitosan etc. for foods rich in unsaturated fats like fish which otherwise make them prone to lipid oxidation causing rancidity, off flavours, hence limiting shelf life. Where TBARS gives secondary oxidation product value, POV gives primary lipids oxidation values for food over storage period [58]. Present study indicated an increasing trend in peroxide value with the passage of time in all the treated groups and control as well. However, all the treatment groups showed significantly better performance than control group (Table\\u0026nbsp;\\u003cspan refid=\\\"Tab5\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003e). But when thyme oil and chitosan coexisted in combination they performed best among all treated groups. This trend of increase in POV over time, may be attributed to the presence of several oxidation promoters in meat for example pro-oxidants like pigments containing hem or non-hem iron such as hemoglobin and myoglobin [59]. Similarly, percentage of lipid and protein also contribute towards oxidation promotion [60]. Comparable results were observed by Nawaz et al. [23] when rosemary essential oil was combined with chitosan to prevent oxidation of lipids. Chitosan formulated lemon extract and essential oil on rainbow trout also resulted in lower POV of samples treated compared to control [54]. In accordance with our work, a low POV was observed in salmon fillets coated in bioactive coating and cinnamon essential oil group demonstrating a clear control on oxidative stress by the phenolic compounds of essential oil and barrier provided by bioactive coating [61]. Multiple mechanisms are engaged by the phenolic compounds present in essential oils to alleviate oxidative stress. These mechanisms include controlling the first phase of oxidation to propagative phase by radical scavenging chain terminating steps through metal chelation and ROS prohibition [62, 18, 50].\\u003c/p\\u003e \\u003cp\\u003eConsumer acceptance and preference can be judged effectively on the basis of sensory analysis of stored and preserved food especially perishable ones like fish. Texture is the key indicator of quality under storage conditions [63]. Assessment of moisture and tenderness has been widely used to predict the texture of fish shelf life extension. In this regard panel of experts revealed that texture decline occurred in control as well as all the treatment groups over the period of storage. But that textural decline in all treatment groups was significantly lower compare to control (Table\\u0026nbsp;7). In terms of juiciness and tenderness, the highest score for texture was observed in combination of thyme essential oil and chitosan group, which may be attributed to hygroscopic and film-forming properties of chitosan and antioxidant properties of thyme oil. Similar findings were reported by Yu et al. [44] for textual stability of mutton patties using thyme essential oil and microencapsulation. Odor assessment by the panel of experts revealed the same pattern as that observed in case of texture. Control group got least score at the end of the storage period that is 21st day whereas thyme oil and chitosan combination group got the highest acceptability score (Table\\u0026nbsp;7). In consistence with our results Nawaz et al. [23] reported least acceptability of control and better score for rosemary essential oil and chitosan coated mori samples. Similarly, Li et al. [64] and Yu et al. [44] also reported similar patterns of results using microencapsulated clove and thyme essential oil, respectively. Microbial control by thyme oils is well established, antimicrobial nature and additive control in a synergistic manner by chitosan was effectively performed which resulted in odor inhibition and increased acceptability of samples. Furthermore, oxidative damage which also contributes to off flavours as well as off odor, was also reduced by the combination group as depicted by lower values of TABARS and TVBN [40, 44]. Colour changes occurred in all samples including control and treatment groups with significantly better colour preservation in treatment groups compare to control and highest score being obtained by combination group (Table\\u0026nbsp;7). Similar results were obtained by Yu et al. [44] where higher colour preservation score was observed in microencapsulated thyme oil treated mutton parties compared to other group and control. Gum Arabic in Yu et al. [44] case seems to work similar as that of edible coating to minimize the volatility of phenolic compounds of thyme oil so that it may work synergistically against the deteriorative factors which cause colour change for example oxidative stress. Least score for firmness in case of control and highest score in case of combination group at the end of the storage period reveals that factors like prevention of autolysis, muscle protein degradation and lipid oxidation might have retained firmness level by the synergistic action of both the components.\\u003c/p\\u003e \\u003cp\\u003eBacteria are very important microbial agents involved in microbial degradation of meat and posing foodborne diseases based public health threat. The utilization of thyme oil as natural preservative to restrict microbial activities is well reported [65]. In our study, an increase in CFU/g was observed during the preservation from day 0 to day 21st in all treated groups. In case of control group it was high (2\\u0026ndash;8.43 log CFU/g) but the treated groups showed comparatively less increase and the range of TVC values for thyme oil, chitosan and combination of thyme oil and chitosan treatment groups, was observed 2 to 6.2 log CFU/g with least values (5.9 log CFU/g) for the combination of thyme oil and chitosan (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e). Marino et al. [66] demonstrated antimicrobial activity of thyme oil against nine Gram-negative and six Gram-positive bacterial strains among which \\u003cem\\u003eEscherichia coli\\u003c/em\\u003e O157:H7 was found most sensitive strain. According to ICMSF, 1986 the upper acceptable limit of TVC is 6\\u0026ndash;7 log CFU/g. The current study showed that TVC values in all treated groups were in acceptable range (2\\u0026ndash;6.2 log CFU/g) from day 0 to day 21.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec21\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eRecommendations\\u003c/h2\\u003e \\u003cp\\u003eIn this study best performance was showed by thyme oil in combination with chitosan. The estimated cost of treatment in combination was Rs. 128/Kg fish meat. This minimal cost addition would not only extend the shelf life of fish meat up to two weeks but also ensure food safety during storage and minimize loss due to spoilage.\\u003c/p\\u003e\\u003c/div\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eFunding declaration\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis study was not funded\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthics statement\\u0026nbsp;\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe fresh specimens of \\u003cem\\u003eLabeo rohita\\u003c/em\\u003e were obtained from a licensed commercial fish market, and no live animals were subjected to experimental procedures.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003ch2\\u003eAuthor Contribution\\u003c/h2\\u003e\\u003cp\\u003eA.S. conducted all experimental work, data compilation and analysis, wrote the manuscript. A.M.K. designed the research work, supervised all research work and data analysis, edited and reviewed the manuscript\\u003c/p\\u003e\\u003ch2\\u003eAcknowledgement\\u003c/h2\\u003e\\u003cp\\u003eWe acknowledge Dr. Syed Zakir Hussain Shah, Department of Zoology, University of Gujrat Pakistan, for his kind consideration of lab access during this research work.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eWHO. The state of food security and nutrition in the world 2018. Building climate resilience for food security and nutrition. Food and Agriculture\\u003cem\\u003e \\u003c/em\\u003eOrganization of the United States of America. 2018.\\u003c/li\\u003e\\n\\u003cli\\u003eRumpa R, Haque M, Alam M, Rahamatullah S.. Growth and production performance of carps in shaded pond in Barisal, Bangladesh. 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Essential Oil and Thymol on the Microbiological Properties of Meat and Meat Products: A Review, \\u003cem\\u003eHeliyon\\u003c/em\\u003e, 8, e10812 https://doi.org/10.1016/j.heliyon.2022.e10812\\u003c/li\\u003e\\n\\u003cli\\u003eMarino, M., Bersani, C., \\u0026amp; Comi, D.G. (1999). Antimicrobial Activity of the Essential Oils of \\u003cem\\u003eThymus vulgaris L\\u003c/em\\u003e. Measured Using a Bioimpedometric Method. \\u003cem\\u003eJournal of Food Protection, \\u003c/em\\u003e62(9), 1017\\u0026ndash;1023.\\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":false,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"food-science-of-animal-resources\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"\",\"sideBox\":\"Learn more about [Food Science of Animal Resources](https://link.springer.com/journal/44463)\",\"snPcode\":\"44463\",\"submissionUrl\":\"https://submission.springernature.com/new-submission/44463/3?\",\"title\":\"Food Science of Animal Resources\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Open\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Thyme oil and chitosan, Natural preservatives, Labeo rohita, Shelf-life extension, Preservation techniques\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-9124794/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-9124794/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eThe role of fishery in economy of a country with agricultural potential like Pakistan is very important. Fish meat has large market share as an important food source with high quality proteins, vitamins and minerals. Rohu (\\u003cem\\u003eLabeo rohita\\u003c/em\\u003e) demand has increased due to consumer preference, market value and high yield in local environment. The huge economic loss due to spoilage at local and commercial level can be minimized by improving meat storage. In recent years the importance of natural organic preservatives has increased due to hazardous effects of synthetic preservatives. In this context essential oils in combination with chitosan have been proved a good edible meat preservative candidate. Keeping in view the importance of natural preservatives, the study was carried out to evaluate the preservation efficacy of thyme oil and chitosan separately and in combination on Rohu (\\u003cem\\u003eLabeo rohita\\u003c/em\\u003e) meat preservation in refrigerated conditions (4℃). Following two-factor factorial completely randomized experimental design (CRD) was used. Thyme oil and chitosan as 0% \\u0026amp; 1% in 2\\u0026times;2 interaction was applied on refrigerated fillets of Rohu meat. The treatment lasted 21 days and sampling was done on day 0, 7, 14 and 21 for analyses. Subsequently, results demonstrated that application of thyme oil (1%) in combination with chitosan (1%) extended shelf life of Rohu meat up to two weeks with maintained sensory acceptability. Hence, our study suggested the thyme oil and chitosan as promising edible organic preservative candidates for preservation of Rohu fillets in refrigerated conditions.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Shelf life extension and meat quality enhancement of Labeo rohita during storage at refrigerated conditions using thyme oil and chitosan as natural preservatives\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2026-05-11 12:40:48\",\"doi\":\"10.21203/rs.3.rs-9124794/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2026-05-04T04:37:18+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2026-03-17T16:30:05+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2026-03-17T09:41:10+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Food Science of Animal Resources\",\"date\":\"2026-03-14T19:54:45+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"food-science-of-animal-resources\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"\",\"sideBox\":\"Learn more about [Food Science of Animal Resources](https://link.springer.com/journal/44463)\",\"snPcode\":\"44463\",\"submissionUrl\":\"https://submission.springernature.com/new-submission/44463/3?\",\"title\":\"Food Science of Animal Resources\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"stoa\",\"reportingPortfolio\":\"Springer Open\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"f78c6314-d9cc-4777-b29d-b5d4dc746289\",\"owner\":[],\"postedDate\":\"May 11th, 2026\",\"published\":true,\"recentEditorialEvents\":[{\"type\":\"reviewersInvited\",\"content\":\"7\",\"date\":\"2026-05-04T04:37:18+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"under-review\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2026-05-11T12:40:49+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2026-05-11 12:40:48\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-9124794\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-9124794\",\"identity\":\"rs-9124794\",\"version\":[\"v1\"]},\"buildId\":\"XKTyCvWXoU3ODBz1xrDgd\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}