LWT - Food Science and Technology 87 (2018) 234e240

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LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

Effect of ultra-high pressure treatment on shucking and meat

properties of red swamp crayfish (Procambarus clarkia)
Ying Shao a, c, Guangquan Xiong a, Jiangang Ling b, Yaqin Hu c, Liu Shi a, Yu Qiao a,
Jingfen Yu b, Yan Cui b, Li Liao a, Wenjin Wu a, Xin Li a, Anzi Ding a, Lan Wang a, *
a
Institute of Agricultural Products Processing and Nuclear- Agricultural Technology, Hubei Academy of Agricultural Sciences, Farm Products Processing
Research Sub-center of Hubei Innovation Center of Agriculture Science and Technology, Wuhan 430064, China
b
Institute of Agricultural Products Processing, Ningbo Academy of Agricultural Sciences, Wuhan 430064, China
c
National Engineering Laboratory of Intelligent Food Technology and Equipment, Key Laboratory for Agro-Products Postharvest Handling of Ministry of
Agriculture, Key Laboratory for Agro-Products Nutritional Evaluation of Ministry of Agriculture, Zhejiang Key Laboratory for Agro-Food Processing, Fuli
Institute of Food Science, College of Biosystems Engineering and Food Science Zhejiang University, Hangzhou 310058, China

a r t i c l e i n f o a b s t r a c t

Article history: The objective of this study was to investigate the effect of ultra-high pressure treatment (0, 100, 200, 300,
Received 27 February 2017 400, and 500 MPa, respectively) on shucking (crayfish meat yield) and meat properties (moisture con-
Received in revised form tent, thermal characteristics, texture properties, protein pattern and microstructure) of red swamp
28 July 2017
crayfish (Procambarus clarkia). Comparing to the control, the crayfish meat yield increased and the
Accepted 29 July 2017
Available online 30 August 2017
moisture content decreased for the samples with the high pressure treatments, however no significant
difference was observed when the pressure above 200 MPa. The denaturation enthalpy (△H) of three
endothermic peaks normally found at 50
C, 65
C, and 80
C were all significantly shifted to lower value
Keywords:
HPP
(p < 0.05) after pressure treatment. With the increase of treated pressure, the hardness, springiness and
Crayfish chewiness values were increased sharply at first and then decreased slightly but all higher than those of
Meat yield the control. Microstructure changed obviously with the increase of the ultra-high pressure. Overally,
Texture 200 MPa was considered as the optimum treatment condition for crayfish shucking without damage the
Histology food quality. The research would provide a reliable reference for ultra-high pressure application in
shucking of red swamp crayfish.
© 2017 Elsevier Ltd. All rights reserved.

1. Introduction In order to meet demands in the seafood industry, red swamp

Red swamp crayfish (Procambarus clarkia) is one of the most
popular and important commercial freshwater species in China,
especially in the southeast regions. Introduced from America (Li,
Deng, Yang, & Wang, 2012), red swamp crayfish has been widely
artificially farmed in inland China, and the total production reached
659,611 tons in 2014 (Lan et al., 2016). In general conditions, most
adults are approximately 5.6 cme11.9 cm in length and resemble
small lobsters (Ding et al., 2012). Red swamp crayfish are highly
appreciated by consumers on account of their high nutritive value
and great flavor when well cooked, and a significant mark has
developed due to convenient breeding, excellent yield, and high
growth rate.
* Corresponding author. Institute for Farm Products Processing and Nuclear-
Agricultural Technology, Hubei Academy of Agricultural Science. China.
E-mail address: lilywang_2016@163.com (L. Wang).
crayfishes are usually shucked so that they can be sold as ready raw
meats with higher consumer acceptance. Manual shucking is a
widely used method, however, it probably impairs red swamp
crayfish meat even when carried out by high-skilled persons,
leading to quality deterioration (Yi et al., 2013). Meanwhile, the
crayfish meat yield is substantially reduced. In addition, the low
efficiency of manual shucking can not be neglected. Nowadays, a
large number of other shucking methods for bivalve shellfish have
been developed to assist manual shucking, such as freezing process
and thermal methods for the raw materials. Martin, Supan, Theriot,
and Hall (2007) have suggested that yielded meat detachments of
Crassostrea virginica with a combined shucking treatment of low-
pressure steam and cryogenic N2/CO2 or ice water could exceed
85%. Nevertheless, automated shucking machines are unlikely to
sever the adductor muscle precisely so that their utilization is
heavily limited. Furthermore, the shucking with heat treatment
will result in the coarse meat quality and the unfavorable taste.

http://dx.doi.org/10.1016/j.lwt.2017.07.062
0023-6438/© 2017 Elsevier Ltd. All rights reserved.
 Y. Shao et al. / LWT - Food Science and Technology 87 (2018) 234e240
Therefore these processes cannot meet consumer demands which
requires seafood to be raw or minimally-cooked (Yi et al., 2013).
Nowadays, ultra-high pressure has been widely employed in
efficient food processing, including meat products, shellfish
shucking, and fruit and vegetable juices processing (Cao et al.,
2012). Ultra-high pressure could disrupt protein conformation
and inter-/intra-molecular interactions, including hydrogen bonds,
electrostatic bonds, hydrophobic interactions as well as disulphide
bonds, resulting in the structural change and the functional
modification, such as protein solubilization, aggregation, denatur-
ation, gelation, or depolymerisation (Aertsen, Meersman,
Hendrickx, Vogel, & Michiels, 2009). Studies have reported that
major changes in the quaternary structure, the tertiary structure,
and secondary structure generally take place at mild pressure
(150 MPa - 200 MPa), moderate pressure (above 200 MPa), and
relatively high pressure (300 MPa - 700 MPa), respectively (Aertsen
et al., 2009). Proposed as a promising alternative for oyster
shucking, ultra-high pressure treatment has several advantages,
including higher muscle integrity, increased meat yield, and
enhanced safety due to the inactivation of enzymes and microor-
ganisms with the minimum loss of color, taste, juicy, and nutri-
tional content (Cruz-Romero, Kelly, & Kerry, 2007). Thus the
physiochemical and functional properties of seafood products are
improved to a certain extent with ultra-high pressure treatment.
Cruz-Romero et al. (2007) have compared three different treat-
ments (ultra-high pressure, cool pasteurization and traditional
pasteurization) to the manual shucking, it turned out that the
treatments had higher oyster shucking yield and cost less labor
than pure manual shucking, and ultra-high pressure treatment was
better than other treatments (the shucking yield of ultra-high
pressure, cool pasteurization and traditional pasteurization treat-
ment was 15.5, 12.5 and 2.6 g/100 g, respectively).
In current, studies on the shucking and meat properties of the
red swamp crayfish by ultra-high pressure have not been done.
Therefore, the objective of this study was to evaluate the effect of
ultra-high pressure treatment on shucking and meat properties of
red swamp crayfish, including moisture content, crayfish meat yield,
thermal characteristics, textural profile analysis (TPA), protein
pattern (SDS-PAGE), and microstructure (histology). The research
would provide a reliable theoretical basis and practical guidance for
ultra-high pressure application in shucking of red swamp crayfish.
2. Materials and methods
2.1. Materials
Fresh red swamp crayfishes (Procambarus clarkii) (approxi-
mately 10 cme11 cm in length, 40 g - 45 g in weight) were pur-
chased from a local fresh supermarket (Wuhan, China). They were
kept in crushed ice and transported to the lab within 30 min. Then
they were cleaned with pre-cooling water prior to further
treatment.
HPP.L2-600/2 ultra-high pressure processor, DSC 200F3 differ-
ential scanning calorimeter, TA-XT2 Texture Analyzer, Gel Doc XR
gel imaging, and Eclipse Ci microscope were provided by Tianjin
Huatai-Senmiao Bioengineering and Technology Co., Ltd (Tianjin,
China), NETZSCH Scientific Instrument Co., Ltd (Selb, Germany),
Stable Micro Systems Ltd (Goldaming, UK), Bio-Rad Laboratories Inc
(Hercules, America), and Nikon Instrument Inc (Tokyo, Japan),
respectively.
2.2. Preparation of samples
2.2.1. Ultra-high pressure treatment
Each fresh red swamp crayfish was packaged into a vacuum bag
235
and then processed in sets of ten per batch. They were vacuum
sealed and placed in an ultra-high pressure processor (HPP.L2-600/
2, Tianjin, China). The pressure inside the processor was increased
at a speed of 5 MPa/s to reach the aim pressure (high hydrostatic
pressure, 100, 200, 300, 400, and 500 MPa, respectively), and held
for 5 min, and finally released within 5 s. The temperature inside
the processor during the processing was slightly raised from 25
C
to 28
C, monitored by a thermometer. Samples without ultra-high
pressure treatment were set as the control. After the ultra-high
pressure treatment, the red swamp crayfish was shucked manually.
2.2.2. Extraction of myofibrillar protein (MP)
Fresh red swamp crayfish meat was minced and homogenized
and then myofibrillar protein (MP) was extracted according to the
method described by Xiong, Cheng, Ye, and Cai (2009) using
20 mmol/L Tris-maleate buffer (pH 7.0) respectively containing
50 mmol/L potassium chloride (KCl) and 0.6 mol/L KCl. The mixture
was centrifuged at 9000 g for 10 min and 30 min, respectively. The
final supernatant was the MP solution.
2.3. Moisture content
Moisture content of red swamp crayfish meat was determined
by the oven drying at constant temperature of 105
C (±5
C) for 2 h
(Juang, Chang, Sung, & Su, 1984). The measurements were taken 3
times.
2.4. Crayfish meat yield
Crayfish meat yield of red swamp crayfish treated under ultra-
high pressure were performed according to the method of Kim,
Meyers, and Godber (1996) with some modifications. Each group
treated with different level of ultra-high pressure contained ten red
swamp crayfishes. The measurements were taken10 times. Crayfish
meat yield was calculated from the following equation:
hX i.
Crayfish meat yield ð%Þ ¼ mi =Mi N  100ði
¼ 1; 2; 3::::::N; N ¼ 10Þ
where Mi was the weight of red swamp crayfish before shucking
while mi was the weight of the shucked crayfish meat.
2.5. Differential scanning calorimetry (DSC)
Thermal properties of the meat of red swamp crayfish treated
under ultra-high pressure were performed by DSC according to the
method of Meng and Ma (2001) with some modifications. Red
swamp crayfish samples of 19 mge25 mg were accurately weighed
into standard aluminum pans and then hermetically sealed. An
empty pan was set as the reference. Samples were heated from
20
C to 100
C at a rising rate of 5
C/min in a differential scanning
calorimeter (TA Instruments, NETZSCH, DSC 200F3, Germany). The
cooling mode was set as mechanical refrigeration. The flow rate of
sweeping gas and protecting gas, both N2, were 20 mL/min and
60 mL/min, respectively. The measurements were repeated 5 times.
Statistical analysis was performed by Proteus Thermal Analysis
(Version 6.1.0, NETZSCH, Germany).
2.6. Textural profile analysis (TPA)
Texture measurements of the second abdominal segment of
crayfish meat were performed using a TA-XT2 Texture Analyzer
(Stable Micro Systems Ltd., Surry, UK) according to the method of
Truong, Buckow, Nguyen, and Stathopoulos (2016). The compression
236 Y. Shao et al. / LWT - Food Science and Technology 87 (2018) 234e240

force axially at 50% strain in a two-cycle compression test was ob-
tained with a P/36R probe. The samples were measured with a 5 g
trigger force at a pretest speed of 0.5 mm/s and a test speed of
0.5 mm/s. TPA parameters, including hardness (N), springiness (mm),
and cohesiveness, were acquired by the Texture Expert software and
recorded. The TPA measurement was conducted in 8 times.
2.7. SDS-polyacrylamide gel electrophoresis (SDS-PAGE)
SDS-PAGE was performed following the procedure described by
Laemmli (1970) using a separating gel of 10% acrylamide and a
stacking gel of 4% acrylamide. Samples (1 mg/mL, 10 mL) were
loaded per lane and the final gels were photographed by a gel
imaging (Gel Doc XR, Bio-Rad, America). The measurements were
taken 3 times.
2.8. Histology
Histology was performed following the procedure reported by
Bahuaud et al. (2008). After the dehydration, paraffin embedding,
sectioning, and dyeing treatment, the microstructure of samples
was observed by the optical microscope (Eclipse Ci, Nikon, Tokyo,
Japan) and collected for the further analysis. The magnification of
scanned images was adjusted to 1500 times. The measurements
were taken 3 times.
2.9. Statistical analysis
The measurements of all the parameters were repeated at least
3 times. The experimental data were calculated by Microsoft Office
Excel 2013 (Microsoft Inc., USA) and presented as Mean ± SD.
Statistical analysis was performed by ORIGIN (Version 8.0; Micro-
soft Software Inc., Northampton, MA) and compared by a two-way
analysis of variance (ANOVA) followed by Duncan's multiple-range
test from the SAS software (Version 8.0; SAS Institute Inc., Cary, NC,
USA). The statistical significance was identified at the 95% confi-
dence level (p < 0.05).
3. Results and discussion
3.1. Moisture content and crayfish meat yield
As shown in Table 1, the moisture contents of red swamp
crayfish treated with ultra-high pressure were significantly
decreased (p < 0.05) comparing with the control (without pressure
treatment) and the crayfish meat yield of red swamp crayfish
treated with ultra-high pressure was significantly higher compared
to control (p < 0.05). The highest moisture content of crayfish was
achieved at 100 MPa (78.78 g/kg) in ultra-high pressure treated
samples. There showed no difference (p > 0.05) among higher
pressure treatments at 200, 300, 400 and 500 MPa. The highest
meat yield was achieved at 200 MPa (22.40 g/100 g), but no sig-
nificant difference was observed among samples treated with 200,
300, 400, and 500 MPa. Presumably pressure treatment lead to the
decline of water holding capacity of muscle protein, contributing to
the penetration of the water embedded in muscle into the extra-
myofibrillar space (Yi et al., 2013). In addition, water loss may be
caused by myfibrillar denaturation (Cruz-Romero, Kerry, & Kelly,
2008). However, no further decrease of moisture content was
observed for samples treated with higher pressure (200 MPa -
500 MPa). Moisture loss might have reached its maximum limit,
and the ultra-high pressure treatment may also prompt water ab-
sorption by strengthening hydration of muscle proteins (Briones-
Labarca, Perez-Won, Zamarca, Aguilera-Radic, & Tabilo-Munizaga,
2012), thus the whole complex system was finally in the status of
equilibrium. Basically consistent with the result in Table 1, Murchie
et al. (2005) have reported that yield of the oyster increases by 25
or 50% with high pressure treatment meanwhile its meat will
recover great appearance, shape, and juiciness.

Table 1
3.2. Differential scanning calorimetry (DSC)
Effect of different ultra-high pressures on the moisture content and meat yield of red
swamp crayfish.
Thermal characteristics of red swamp crayfish meat for both
Ultra-high pressure/MPa Moisture content/g/100 g Meat yield/g/100 g
untreated and treated with different ultra-high pressure (100, 200,
0 81.21 ± 0.93a 12.30 ± 0.83c 300, 400, and 500 MPa, respectively) around 50
C, 65
C, and 80
C
100 78.78 ± 0.13b 17.55 ± 0.41b
are depicted in Table 2. Three endothermic peaks observed at peak
200 75.84 ± 0.42c 22.40 ± 0.11a
300 75.79 ± 0.06c 21.99 ± 0.04a temperatures of 55
C ± 5
C, 65
C ± 5
C, and 75
C ± 5
C
400 75.96 ± 0.38c 21.17 ± 0.13a represent myosin, a combination of sarcoplasmic and connective
500 75.17 ± 0.12c 22.00 ± 0.02a tissue proteins, and actin, respectively (Khan, Ali, Abid, Ahmad,
Note: Different lowercases letters in the same column mean significant difference et al., 2014), which is consistent with the result in Fig. 1. The
(p < 0.05) (n (moisture content) ¼ 3; n (crayfish meat yield) ¼ 10). denaturation temperature and the denaturation enthalpy of

Table 2
Effects of different ultra-high pressures on the thermal characteristics of red swamp crayfish around 50
C, 65
C and 80
C.

Ultra-high pressure/MPa 0 100 200 300 400 500

50
C To/
C 44.2 ± 0.3a 44.6 ± 0.2a 38.3 ± 0.5bc 38.8 ± 0.4b 37.9 ± 0.5c 37.8 ± 0.4c
Tp/
C 47.5 ± 0.3ab 48.0 ± 0.4a 42.0 ± 1.7c 44.9 ± 3.3b 39.4 ± 1.2c 39.7 ± 0.8c
Tc/
C 50.0 ± 0.4a 50.7 ± 0.6a 46.2 ± 2.0b 48.7 ± 1.9a 42.6 ± 1.9c 42.4 ± 0.6c
△H/(J/g) 0.94 ± 0.03a 0.91 ± 0.03a 0.29 ± 0.03b 0.28 ± 0.10b 0.26 ± 0.04b 0.21 ± 0.00b
65
C To/
C 63.4 ± 0.9ab 61.9 ± 0.6b 61.8 ± 1.2b 64.9 ± 2.8a 61.5 ± 0.3b 63.4 ± 1.0ab
Tp/
C 66.5 ± 0.2a 66.1 ± 0.4ab 64.1 ± 3.1c 66.4 ± 1.1a 65.2 ± 0.7abc 64.3 ± 0.6bc
Tc/
C 70.6 ± 0.3a 70.6 ± 0.7a 66.8 ± 3.0b 68.0 ± 2.6ab 66.6 ± 1.9b 66.5 ± 2.0b
△H/(J/g) 0.46 ± 0.04a 0.44 ± 0.06a 0.12 ± 0.03b 0.11 ± 0.03b 0.09 ± 0.03b 0.08 ± 0.01b
80
C To/
C 77.8 ± 4.2ab 77.2 ± 0.8b 80.0 ± 0.8ab 81.8 ± 3.2a 80.1 ± 0.6ab 78.0 ± 0.9ab
Tp/
C 81.9 ± 0.2bc 81.1 ± 0.4c 82.7 ± 0.1ab 83.6 ± 0.9a 83.2 ± 0.8a 81.3 ± 0.8c
Tc/
C 85.5 ± 0.5ab 83.0 ± 1.9b 84.2 ± 1.5ab 86.1 ± 0.7a 85.3 ± 1.4ab 84.2 ± 1.2ab
△H/(J/g) 0.29 ± 0.05a 0.26 ± 0.05a 0.14 ± 0.03b 0.13 ± 0.02b 0.12 ± 0.04b 0.13 ± 0.00b

Note: To, Tp, Tc, and △H indicate the original temperature, peak temperature, conclusion temperature, and denaturation enthalpy of red swamp crayfish, respectively.
Different lowercases letters in the same row mean significant difference (p < 0.05) (n ¼ 5).
 Y. Shao et al. / LWT - Food Science and Technology 87 (2018) 234e240 237

Fig. 1. Differential Scanning Calorimetry (DSC) curve of red swamp crayfish meat. a:
The endothermic peak around 50
C; b: The endothermic peak around 65
C; c: The
endothermic peak around 80
C.

endothermic peaks in the DSC curve are positively correlated to the

protein stability, especially the MP (Zhou, Benjakul, Pan, Gong, &
Liu, 2006). No significant difference (p > 0.05) is observed be-
tween the control and the sample treated with 100 MPa (Table 2),
which meant that ultra-high pressure at low level (100 MPa) had no
obvious effect on thermal characteristics of red swamp crayfish.
However, thermal parameters differed significantly (p < 0.05) when
the ultra-high pressure reached 200 MPa, especially the peak
temperature (Tp) and the denaturation enthalpy (△H). The dena-
turation temperatures of peaks around 50
C and 65
C both
showed a downtrend while that around 80
C showed an uptrend
on the whole as the ultra-high pressure increased. The denatur-
ation enthalpy of peaks around 50
C, 65
C, and 80
C all decreased
significantly (p < 0.05) as the ultra-high pressure increased from
100 MPa to 200 MPa and then showed no significant change
(p > 0.05) as ultra-high pressure further increased. The reduction in
the volume of muscle proteins induced by the ultra-high pressure
has a negative effect on ionic and hydrophobic interactions in the
protein network (Khan, Ali, Abid, Cao, et al., 2014). Protein struc-
Fig. 2. Effect of ultra-high pressure on texture of red swamp crayfish meat. (a):
tures will change heavily due to the interdependent antagonistic-
hardness; (b): springiness; (c):cohesiveness. Note: Different lowercases letters mean
like effect caused by ultra-high pressure on the protein matrix, significant difference (p < 0.05) (n ¼ 8).
which finally lead to the protein denaturation (Ferna
ndez-Martín,
Otero, Solas, & Sanz, 2000). Cao, Xia, Zhou, and Xu (2012) have
reported ultra-high pressure could contribute to the complete
denaturation of myosin, which is consistent with the result in
Table 2. The less stable sarcoplasmic and connective tissue proteins
under the condition of ultra-high pressure treatment are due to
weakened hydrogen bonds within their triple a-helix structure
(Ferna
ndez-Martín et al., 2000). However, the relatively slighter
effect of the ultra-high pressure on actin is observed for its
comparatively lower proportion in muscle protein.

3.3. Textural profile analysis (TPA)

TPA parameters of red swamp crayfish untreated and treated

with different ultra-high pressure (100, 200, 300, 400, and
500 MPa, respectively) were shown in Fig. 2. Hardness, springiness,
and cohesiveness of samples all increased compared to the un-
treated control. Specifically, Cohesiveness showed a slight decrease
from 0 to 200 MPa (p > 0.05), a dramatically sharp increased from
200 MPa to 400 MPa (p < 0.05) and finally a weak decreased from
400 MPa to 500 MPa (p > 0.05).
Textural characteristics greatly affect consumer acceptability thus
playing an important role in the manufacture of refined processed
meat-based products (Chan, Omana, & Betti, 2011). High pressure Fig. 3. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) profile of MP extracted
from red swamp crayfish untreated and treated with different ultra-high pressures
strengthens interactions between matrix components and alters the (100, 200, 300, 400, and 500 MPa, respectively). A: Untreated (control); B, C, D, E and F:
water distribution so textural properties are improved to some Treated with 100, 200, 300, 400, and 500 MPa, respectively. MHC: Myosin heavy
extent (Cruz et al., 2010; Picouet et al., 2012). It has been suggested chains; MLC: Myosin light chains.
238 Y. Shao et al. / LWT - Food Science and Technology 87 (2018) 234e240

that texture is correlated with the binding strength (Herrero et al.,
2007). At certain pressure levels, the formation of hydrogen bonds
and non-disulphide covalent bonds prompted denser protein gel
networks causing protein molecules to be arranged more closely
(Chanarat & Benjakul, 2013; Xia et al., 2017), contributing to the
increased textural characteristics. Crehan, Troy, and Buckley (2000)
report that binding strength and texture parameters are improved
at lower pressure levels (<150 MPa), which was similar to the result
depicted in Fig. 2. However, hardness and springiness both increased
significantly from 0 to 200 MPa (p < 0.05) and decreased slightly
from 200 MPa to 500 MPa (p > 0.05), which was in accordance with
the report of Truong et al. (2016). The progressively increasing ultra-
high pressure might result in the denaturation, aggregation or
gelation of muscle proteins due to disintegrated muscle fibers, dis-
rupted protein conformation, the ruptured protein networks
(Truong et al., 2016). Lachowicz, Sobczak, Gajowiecki, and Zych _

Fig. 4. Microstructure of red swamp crayfish untreated and treated with different ultra-high pressures (100, 200, 300, 400, and 500 MPa, respectively). (a): Cross sectional
microstructure; (b): Vertical sectional microstructure. A: Untreated (control); B, C, D, E and F: Treated with 100, 200, 300, 400, and 500 MPa, respectively.
 Y. Shao et al. / LWT - Food Science and Technology 87 (2018) 234e240
(2003) have reported the disintegration of muscle fibers could be
reflected in the reduction of their cross-sectional area as well as the
peri- and endomysium thickness. In addition, ultra-high pressure
causes the loss of protein functionality, particularly myosin heavy
chains, leading to final textural changes (Briones-Labarca et al., 2012;
Cheftel & Culioli, 1997), which will heavily affect the processing of
red swamp crayfish.
3.4. SDS-PAGE
The protein pattern of MP extracted from red swamp crayfish
untreated and treated with different ultra-high pressure (100, 200,
300, 400, and 500 MPa, respectively) was shown in Fig. 3. The in-
dividual soluble protein bands of MP were characterized by the
molecular weight (Mw) distribution of the standard protein marker
in SDS-PAGE pattern under non-reducing conditions (Grossi et al.,
2016). Typical profile for MP is presented including two major
protein bands myosin heavy chain (MHC, 220 kDa) and actin
(44.3 kDa) and several low intensity protein bands consisting of
paramyosin (100 kDa), tropomyosin (36 kDa), troponin (33 kDa),
and myosin light chains (MLC, 15, 17, and 24 kDa, respectively). The
intensity of almost all bands of MP showed an obvious increase
when the red swamp crayfish was treated under 100 MPa
compared to the untreated control. It is speculated that ultra-high
pressure at low level (100 MPa) causes the swelling of MP, which
is then disrupted and dispersed into short filaments (Sikes, Tobin, &
Tume, 2009). The increase of solvent accessibility to the smaller
modified structures results in increased solubilization of myosin,
actin, and some other proteins (Sikes et al., 2009), thus contributing
to the higher intensity protein bands. It was in agreement with the
finding of Grossi et al. (2016) that the ultra-high pressure leads to
the initial denaturation and dissociation of MHC as well as MLC,
followed by the immediate either aggregation or degradation of
MHC. However, the main bands of MP gradually decreased and
disappeared under higher ultra-high pressure levels (200, 300, 400,
and 500 MPa) and were obviously less dense than the control. The
excessive ultra-high pressure treatment may lead to protein
denaturation (Ferna
ndez-Martín et al., 2000) or the formation of
larger aggregates with other proteins into a denser protein network
at higher pressures (Tintchev et al., 2013), inducing the insolubili-
zation of proteins (Chan et al., 2011; Grossi et al., 2016).
3.5. Histology
The cross (a) and vertical (b) sectional microstructures of red
swamp crayfish untreated and treated with different ultra-high
pressure were depicted in Fig. 4. Gels with irregular filaments in
both Fig. 4 (a) and (b) were exhibited in untreated samples. A
denser microstructure containing a small amount of thin strands
was observed as the ultra-high pressure increases to 100 MPa. This
may be due to the increased intra-/inter-actions between muscle
proteins (Cao, Xia, Zhou, & Xu, 2012). Ashie, Simpson, and
Ramaswamy (1997) have also suggested that there would show
the compacting of blue fish muscle fibres with the ultra-high
pressure at 100 MPa. When the ultra-high pressure further
increased to 200 MPa, a reticular structure with a large amount of
thick strands appeared. As has been reported by Gudbjornsdottir,
Jonsson, Hafsteinsson, and Heinz (2010), the space between the
myofibrils (cells) increased with the increase of pressure. However,
the muscle remains compacted (Hurtado, Montero, Borderías, &
Solas, 2001). When the ultra-high pressure reached 300 MPa or
even higher, large fractures were formed. It is postulated that
fractures in muscle fibres are usually attributed to the reversible or
irreversible protein gelation caused by partial protein unfolding
(Ahmed, Ramaswamy, Alli, & Ngadi, 2003; Briones-Labarca et al.,
239
2012). In addition, the formation of a globular aggregation induced
by the cross-links of myosin heavy chains results in the strength-
ened intermolecular bond formation (Cao et al., 2012). Meanwhile,
more thin filaments are observed for the protein denaturation. On
the one hand, higher ultra-high pressure could lead to the partial
unfolding of muscle proteins so that the covalent and non-covalent
interactions are then promoted (Angsupanich, Edde, & Ledward,
1999). On the other hand, breakage and disintegration of the fi-
bres will occur at higher pressures (more than 300 MPa) (Ashie
et al., 1997).
4. Conclusions
The research on red swamp crayfish shucking would be bene-
ficial for developing precooked seafood product and efficient pro-
cessing industry. The ultra pressure treatment resulted in partial
protein denaturation, tightened muscle tissue, increased hardness
and reduced springiness. Treatment at 200 MPa for 5 min is the
optimum for crayfish shucking and maintaining the meat qualities.
This study provides a reliable theoretical basis and a practical
reference for potential application of ultra pressure treatment on
shucking of red swamp crayfishes.
Acknowledge
The authors would like to thank the financial support from Ningbo
Public Welfare Major Program [grant numbers 2015C110002].
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