Evaluation of sausage prepared from spent hen meat with different replacement Levels of spent hen surimi Abstract Breast and leg spent hen surimi were employed in the preparation of sausage

Evaluation of sausage prepared from spent hen meat with different replacement Levels of spent hen surimi

Abstract
Breast and leg spent hen surimi were employed in the preparation of sausage. The sausage was formulated with 10, 20 and 30 % of breast type surimi and leg type surimi and stored at -18 ?C for 5 months. The proximate composition, physicochemical properties, cooking characteristics, microbiological analysis and sensory properties of sausage were evaluated. Sausages formulated with breast type surimi had higher crude protein (15.68%), total volatile basic nitrogen (TVBN) (12.33 mg N/100g), water holding capacity (WHC) (3.96 cm2/0.3g), cooking loss (25.48%) and total yeast and mold counts than sausages formulated with leg type surimi. However, sausages formulated with breast type surimi had lower moisture content(61.88%), thiobarbituric acid (TBA) (0.64mg malonaldehyde/kg), plasticity (2.27 cm2/0.3g), pH (6.24), fat retention (46.26%), water retention (67.52%), total bacterial, proteolytic bacteria, lipolytic bacteria and psychrophilic bacteria counts than sausages formulated with leg type surimi. Sausages formulated with different replacement levels of surimi had higher moisture, carbohydrate, fat retention, water retention plasticity, pH values and sensory properties and lower crude protein, crude fat, total ash, TVBN, TBA, cooking loss, WHC values, total bacterial, proteolytic bacteria, lipolytic bacteria and psychrophilic bacteria and total yeast and mold counts than control sausage. Cooking loss, TVBN, and TBA were decreased as storage time increased, while plasticity and sensory properties had an opposite trend.
Keywords
Surimi, Sausage, Spent hen, Frozen storage, Microbiological quality
1. Introduction
Spent laying hen meat had hard muscle; thus, these meats would like adequate method to enhance their acceptableness (Nowsad et al. 2000 and Ilayabharathi et al. 2012). The emulsion-based product is one such technology which could convert meat of low organoleptic worth into an extremely acceptable product (Ilayabharathi et al. 2012). Sausage could be a product wide consumed worldwide and contains essentially meat and fat (solid phase) spread into ice/water (liquid phase) forming a stable mass that mixed with additives, stuffed into appropriate casings, and heat treatment (Mercadante et al. 2010). Specifically, chicken emulsion sausage could be a known chicken meat-based item. The production and utilization of chicken sausage are increasing at the world from day to day. These sausages are become known because of their sensory attributes and simplicity of preparation (Piotrowicz and Mellado, 2015). Surimi is light in color, has a good odor, low in fat and high in myofibrillar protein and has great useful properties, like gelling, binding, and emulsifying properties (Zhou et al., 2006 and Kim and Park, 2007). Therefore surimi can be added to sausage formula to improve their acceptability. Little knowledge is accessible on the preparation of sausages from spent laying hen meat (Arabi, 2014, Souza et al. 2011 and Singh, 2001) and no reports accessible on the preparation of spent laying hen meat sausages with surimi from spent hen meat. Consequently the target of this work was to prepare sausage from spent laying hen meat with 10, 20 and 30% replacement levels of breast and leg spent hen meat surimi. The proximate composition, physicochemical properties and microbiological analysis of sausage were evaluated throughout frozen storage at -18oC for 5 months.

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2. Materials and methods
2.1. Materials
Two years old chicken females (spent laying hens) weighing 5-6 kg were bought from the native market in Cairo, Egypt. Those chicken might have been slaughtered, permitted to drain to 5 minutes, scalded to 2 min toward 60ºC, plucked by hand, eviscerated, rinsed with tap water, skinned, deboned and at long last trimmed from claiming fat, as needed for the transforming.
Texturized soy was obtained from Food Technology Research Institute, Agriculture Research Center, Giza, Egypt. It was rehydrated with water at a ratio of 1:2 (w/v) and minced twice through 3mm plate. Skimmed milk, spice mixtures, starch, sodium chloride and fresh garlic were bought from the native market in Cairo, Egypt. Sodium tripolyphosphate was obtained from El-Gomhoria Company in Cairo, Egypt.

2.1.1. Preparation of sausage
The sausage was prepared according to the method described by Jin et al. 8. The control sausage was formulated to contain 60% lean chicken meat, 15% beef fat, 10% ice flakes, 6.5% texturized soy, 3% skimmed milk, 1.8 spices mixture, 1.6% starch, 1.5% sodium chloride, 0.5% fresh garlic and 0.1% sodium tripolyphosphate. Sausages containing surimi were prepared by replacing different levels of lean chicken meat (10, 20 and 30%) by equal amounts of the breast type surimi and leg type surimi. Appropriate amounts of each formulation were mixed by hand and subjected to final grinding at 0.4 cm plate. Each formulation was stuffed in the natural mutton casing. Sausages were placed on the foam plate, wrapped with polyethylene film and frozen stored at -18?C for five months. Sausage samples were taken every month for analysis.
2.2. Methods
2.2.1. Proximate composition
Moisture, crude fat, crude protein and total ash contents were determined as described by AOAC (2000). Total carbohydrate content was calculated by difference.

2.2.2. Total volatile basic nitrogen, thiobarbituric acid, and pH values:
Total volatile basic nitrogen (TVBN) and thiobarbituric acid value (TBA) were evaluated as described by Winton and Winton (1958) and Kirk and Sawyer (1991), respectively. The pH value was measured by adding ten percent of chicken sausage sample in distilled water using a Jenway Digital pH meter (Model 3510).

2.2.3. Cooking characteristics:
Cooking loss, fat retention, and moisture retention were determined as described by Bertram et al. (2003) and Alesson-carbonell et al. (2005).
Treatments were weighted before and after cooking. Cooking loss, fat retention, and moisture retention were calculated as follows:

Uncooked sample weight – Cooked sample weight
%cooking loss = ——————————————————————×100
Uncooked sample weight

Cooked sample weight×%fat in the cooked sample
%fat retention= —————————————————————————×100
Uncooked sample weight×%fat in the uncooked sample

Cooked sample weight×%moisture in the cooked sample
%moisture retention= ——————————————————————————-×100
Uncooked sample weight×%moisture in the uncooked sample

2.2.4. Water holding capacity and plasticity:
The procedure of Wierbicki and Deatherage (1958) was used to measure water holding capacity and plasticity.

2.2.5. Bacteriological methods:
2.2.5.1. Total bacteria, psychrophilic bacteria, coliform bacteria and Staphylococcus aureus counts:
The procedures of APHA (1976) were followed to count total bacteria, psychrophilic bacteria, coliform bacteria and Staphylococcus aureus.
2.2.5.2. Lipolytic bacteria and yeast and mold counts:
The procedures as the mansion in Difco manual (1984) were used to count lipolytic bacteria and yeast and mold counts.
2.2.5.3. Salmonella spp:
The method described by FAO (1979) was followed to count Salmonella spp.

2.2.6. Sensory properties:
Sausages were cooked in boiling water at 100oC for 10 min before introduced to panelists. The procedure of Mansour and Khalil (1999) was carried out to evaluate the sensory properties of sausages. The taste, odor, color, texture and overall acceptability were evaluated using a hedonic scale of nine-points for each trait.

2.2.7. Statistical analysis:
Data were analyzed using a Statistical Analysis System (SAS Institute, Inc., Cary, NC, 2008). Data were analyzed by three ways analysis. The acceptable level of probability was 5% for comparison.

3. Results and discussions
3.1. Proximate composition
Proximate composition of sausage was affected (p ? 0.05) by surimi type, surimi level and storage time (Table 1). Sausage formulated with breast type surimi had higher (p ? 0.05) crude protein and lower (p ? 0.05) moisture and crude fat contents than sausage formulated with leg type surimi. These might be due to higher protein content and lower moisture and crude fat contents of breast surimi than leg surimi (Mohammed et al., 2017a). There were no differences (p > 0.05) in total ash and carbohydrate contents between sausage formulated with breast and leg type surimi.
There were significant (p ? 0.05) differences in proximate composition between control sausage and sausages formulated with different replacement levels of surimi. Sausages formulated with surimi had higher (p ? 0.05) moisture and carbohydrate contents and lower (p ? 0.05) crude protein, crude fat and total ash than control sausage. The moisture contents of sausages formulated with surimi were not (p ? 0.05) affected by the levels of surimi. Crude protein, crude fat and total ash contents were significantly (p ? 0.05) decreased by increasing the levels of surimi. However, carbohydrates content were significantly (p ? 0.05) increased by increasing the replacement levels of surimi. Ismail et al. (2014) found that moisture, crude protein and crude fat of sausages containing duck surimi were 58.52, 14.17 and 15.77 %, respectively.
Proximate composition of all sausages was not (p > 0.05) affected by frozen storage up to the second month. From the third month of storage, moisture content was decreased (p ? 0.05), while crude protein, crude fat, total ash, and carbohydrates were increased. The reduction in moisture might be due to the drip loss and evaporation of moisture during storage period (Osheba, 2003 and El-Kordy, 2006). However, the increment in crude protein, crude fat, total ash, and carbohydrates was attributed to the reduction in moisture content during frozen storage. Mohammed et al., (2017a) reported that crude fat, total ash and carbohydrates contents of spent hen type surimi did not significantly (p > 0.05) change until two months of storage followed by an increase up to the end of storage time.

3.2. Total volatile basic nitrogen and thiobarbituric acid
Total volatile basic nitrogen and TBA of sausages were affected (p ? 0.05) by surimi type, surimi level and storage period (Table 2). Sausage formulated with breast type surimi had higher (p ? 0.05) TVBN and lower TBA values than sausage formulated with leg type surimi. Similar observation was reported by Mohammed et al. (2017a) for breast type surimi and leg type surimi. Proximate composition of breast and leg type surimi reported in Table (1) supported these results. However, Ismail et al. (2014) reported that the fat content and TBA values for all sausage samples containing duck surimi-like material substitution had the positive weak correlation (r = 0.144).
Sausage formulated with different replacement levels of surimi had lower (p ? 0.05) TVBN and TBA values than control sausage. Jin et al. (2007) reported that the TBA value was significantly lower in sausage containing spent laying hen surimi sausages than the control sausage. Total volatile nitrogen and TBA values were decreased by increasing the replacement levels of surimi. These could be attributed to washing steps during surimi preparation (Mohammed et al., 2017a). Ismail et al. (2014) found that the use of surimi-like material substitution in sausage was effective in retardation of the lipid oxidation in duck sausages.
Total volatile basic nitrogen and TBA values of sausages were gradually (p ? 0.05) increased by increasing the storage period. Although TVBN and TBA values of sausages were increased with the storage period, their values (7.79- 17.96 mg N/100g and 0.47 – 0.89 mg malonaldehyde/kg) were below the maximum permissible limit (20 mg N/100g and 0.9 mg malonaldehyde/kg) reported by the Egyptian Standard Specifications (2009) for poultry meat products. These results are in agreement with those obtained by Mohammed et al. (2017a) for breast and leg type surimi and Ismail et al. (2014) for sausages containing duck surimi.

3.3. Water holding capacity, plasticity, and pH value
Water holding capacity, plasticity and pH value of sausages were affected (p ? 0.05) by surimi type, surimi level and storage period (Table 3). Sausages formulated with breast type surimi had higher (p ? 0.05) WHC value and lower plasticity and pH values than sausages formulated with leg type surimi. Mohammed et al. (2017a) reported that breast type surimi had a lower pH value than leg type surimi. Sausage formulated with different replacement levels of surimi had higher (p ? 0.05) plasticity and pH values and lower (p ? 0.05) WHC values than control sausage. These could be attributed to the addition of cryoprotectants and washing process during surimi preparation. Water holding capacity values of sausages were (p ? 0.05) decreased and plasticity and pH values increased by increasing the replacement levels of surimi. Agustini et al. (2008) found that the addition of cryoprotectants can improve quality and water holding capacity of surimi.
Plasticity values of sausages were significantly (p ? 0.05) decreased by increasing the storage period. However, the gradual (p ? 0.05) increase in water holding capacity values and pH values of sausages were observed from the second month of storage up to the end of storage period. Ohkuma et al. (2008) reported that the water holding capacity of myofibrillar proteins in surimi are lost because the proteins undergo unfolding and aggregation during processing and storage. Nopianti et al. (2012) found that the WHC of bream surimi treatments decreased as the frozen storage time increased. The increment of pH values might be attributed to the production of volatile basic components such as ammonia and total volatile nitrogen by meat spoilage bacteria (Osheba, 2013).

3.4. Cooking Characteristics
Cooking loss, fat retention and water retention of sausages were affected (p ? 0.05) by surimi type, surimi level and storage period (Table 4). Sausages formulated with breast type surimi had a higher (p ? 0.05) cooking loss and lower fat retention and water retention than sausages formulated with leg type surimi. Sausages formulated with different replacement levels of surimi had lower (p ? 0.05) cooking loss and higher (p ? 0.05) fat retention and water retention than control sausage. These could be attributed to the addition of cryoprotectants during surimi processing. Cooking loss of sausages were gradually (p ? 0.05) decreased and fat retention and water retention gradually (p ? 0.05) increased by increasing the replacement levels of surimi. The water holding capacity of sausages formulated with different levels of breast and leg type surimi reported in Table (3) supported these results. These results are in agreement with Marianski and Marianski (2009) who suggested that the addition of phosphate prevents water loss during cooking.
Cooking loss values of sausages were gradually (p ? 0.05) increased with the storage period. The increment of cooking loss might be attributed to protein denaturation and the loss of protein solubility. However, fat retention and water retention of sausages were not (p > 0.05) affected up to the third month of storage. From the fourth month of storage, fat retention and water retention of sausages were decreased (p ? 0.05).Mohamed (2011) found that the frying loss of chicken burgers was increased by increasing frozen storage period.

3.5. Microbiological analysis
Microbiological analysis of sausage as affected by surimi type, surimi level and storage at -18°C for 5 months are presented in Tables (5). Sausages formulated with surimi had lower total bacterial, proteolytic bacteria, lipolytic bacteria, psychrophilic bacteria and yeast and mold counts than control sausage. The reduction of microorganisms in sausages formulated surimi may be attributed to the washing process during surimi preparation. A similar observation was reported by Mello et al. (2012). Sausages formulated with breast type surimi had lower total bacteria, proteolytic bacteria, lipolytic bacteria and psychrophilic bacteria and higher total yeast and mold counts than leg type surimi. Coliform bacteria, S. aureus, and Salmonella spp were not detected in all formulated sausages. Similar results were reported by Mohammed et al. (2017a and b) who found that coliform bacteria, Staphylococcus aureus and Salmonella spp were not detected in surimi prepared from breast and leg of spent hen meat and kobeba prepared from spent hen meat. The total bacterial and yeast and mold counts of control sausage in the present study ranged from 9.96×104 to 5.62×105and from 4.0×10 to 1.42 ×102, respectively. However, total bacterial and yeast and mold counts of sausages formulated with different replacement levels of surimi ranged from 4.67×104 to 3.71×105 and from not detected to 1.36 ×102, respectively. Gheisari (2011) reported that the maximum permissible counts for total bacteria and yeast and mold for sausage products were 1×105 and 1×102CFU/g, respectively.
Total bacteria, proteolytic bacteria, lipolytic bacteria, psychrophilic bacteria and total yeast and mold counts of sausages were decreased by increasing the levels of breast and leg type surimi. This decrease might be due to the addition of sodium tripolyphosphate during the preparation of surimi, which has antimicrobial activity (Mohammed et al, 2017a). Total bacteria, proteolytic bacteria and lipolytic bacteria counts of sausages were decreased up to the second month of storage followed by gradual increased up to the fifth month of storage. Psychrophilic bacteria counts of sausages showed gradual decreases up to the third month of storage followed by gradual increases up to the fifth month of storage. However, total yeast and mold counts of sausages were gradually decreased as the storage period progressed. The reduction in total microorganism counts during frozen storage might be due to the breakdown of microorganism cell wall by ice-crystals formed during the frozen process (Mohammed et al, 2017a and b). On the other hand, the increase in microorganism counts after reduction period may be due to the increasing simple nitrogen compounds such as amino acids and fatty acids produced by hydrolysis of protein and fat during storage consequently lead to suitable conditions for microorganism growth (Rasha, 2011).

3.6. Sensory properties
Sensory properties of sausages were affected (p ? 0.05) by surimi type except for odor and overall acceptability, surimi level and storage period (Table 6). Sausages formulated with breast type surimi had a higher (p ? 0.05) color and lower taste and texture than sausages formulated with leg type surimi. These could be attributed to the high-fat content in leg type surimi than breast type surimi (Mohammed et al, 2017a). The crude fat contents of sausages formulated with breast and leg type surimi reported in Table (1) supported these results. Sausages formulated with different replacement levels of surimi had higher (p ? 0.05) sensory properties than control sausage. Sausages formulated with 20% replacement levels of surimi had higher (p ? 0.05) rating scores ranged between 7.53 (like moderately) and 8.22 (like very much) for all sensory properties than 10 and 30% replacement levels which ranged between 6.50 (like slightly) and 7.78 (like moderately). Trindade et al. (2005) found that the overall product quality decreased with the increase of spent hen meat percentage in the formulation of sausages; however, the values for this attribute remained higher than the acceptance level five. Jin et al. (2007) indicated that the taste of sausage containing spent laying hen surimi by panels was not much different compared with the original pork sausage.
All sensory properties of sausages were gradually decreased as the storage period increased. During storage, the quality attributes of sausage deteriorate due to lipid oxidation and microbial growth. Kaba (2006) reported that there was a very significant negative relationship between the sensory properties of frozen fish surimi and storage period.

4. Conclusion
From the above results, it could be concluded that spent hen surimi could be incorporated with spent hen meat to produce highly acceptable sausage products. Addition of 20% replacement level of surimi improved cooking and sensory properties of sausage.

Conflict of interests: Authors declare that there’s no conflict of interests relating to the publication of this paper.

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