Luận án tiến sĩ khả năng sản xuất và đa hình gen PRKAG3 của lợn lũng pù và lợn bản

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INTRODUCTION
1. THE RATIONALE OF THE STUDY
Lung Pu and Ban (Hoa Binh Ban) pigs are 2 native breeds which
have a great potential to be exploited/utilized and developed because
of their medium body size, a better productivity compared to that of
other native breeds, a relatively high number of population,
homogenous appearance and especially of their good adoption to
harsh living conditions in high rocky mountainous areas. There have
been several studies on Ban and Lung Pu pigs. However, these
studies were only ended up with an assessment of the origin,
characteristics of appearance and some production traits of these
animals. There has not been any study systematically evaluating
genetic potentials of production traits, especially relationships
between phenotypes and genotypes of meat quality traits of these
breeds.
Due to the above mentioned reality, it is essential to carry out a
research to systematically and adequately evaluate characteristics of
appearance and performance, associated with genetic polymorphisms
of PRKAG3 gene and examine ìnluence of this gene on the quality of
meat of the 2 native pig breeds. The results of this research will
contribute to effective orientations of selection, utilization of genetic
resources of the 2 breeds for enhancement of sustainability of pig
production systems. For all above reason, a research project on
"Performance and polymorphism of PRKAG3 gene of Lung Pu and
Ban pigs" was undertaken
2. OBJECTIVES OF THE STUDY
Evaluate appearance characteristics, perfomance, and influence
of genetic polymorphisms of PRKAG3 gene associated with meat
quality of Lung Pu and Ban pigs for breeding programs and
development orientations of these two pig breeds.

3. SCIENCTIFIC AND PRACTICAL SIGNIFICANCES OF
THE STUDY
3.1. Scientific significance
- The study is the first scientific work systematically dealing
with characterization of appearance, growth and reproduction as well
as meat yield and quality of Lung Pu and Ban pigs.


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- This is also the first scientific work dealing with genetic
polymorphisms of PRKAG3 gene and its influence on some meat
quality parametters (meat color) in Lung Pu and Ban pigs.
- Results of the study can be used as a good reference resource
in research and teaching on genetic resources of indigenous pigs.
3.2. Practical significance
- Results of the study could be a basis for an orientation of
selection to improve quality of Lung Pu and Ban pigs.
- Results of the study on performance could be a scientific basis
for improvement of rearing process of two pig breeds in household
conditions in high mountainous areas.
4. NEW FINDINGS OF THE STUDY
- Full and systematic assessment of characteristics of appearance
from hair coat and skin color, reproduction, growth, carcass yield and
meat quality of Lung Pu and Ban pigs.
- Identifying PRKAG3 genetic polymorphisms including 4
polymorphism : G52S/HphI, T30N/StyI, V199I/BsaHI, R200Q/BsrBI
in the 2 pig breeds: Lung Pu and Ban pig.
- Initial assessment of influences of PRKAG3 genetic
polymorphisms on meat quality of Ban pig. Polymorphisms of
T30N/StyI and G52S/HphI affected the brightness and

polymorphisms of T30N/StyI affected the yellowness of meat.
CHAPTER I
LITERATURE REVIEW
1.1. Appearance characteristic of native pig breeds
There is a variety of indigenous breeds of Vietnam, they are
distributed throughout all the regions of the country with unique
features. It was shown that Vietnam now has about 26 native pig
breeds (Ta Thi Bich Duyen et al., 2013; Nguyen Van Duc, 2012).
There have been several studies on Lung Pu pig (Nguyen Van Duc,
2005; Nguyen Van Duc et al., 2008; Nguyen Van Duc et al., 2012;
Trinh Quang Phong et al., 2009; Trinh Quang Phong et al., 2011) and


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research on Ban pig (Quach Van Thong, 2009; Vu Dinh Ton and
Phan Dang Thang, 2009).
1.2. Reproductive performance of native pig breeds
Most indigenous breeds have the first estrus at an early age, a
low reproductive performance (5-8 piglets/litter). The number of
litters/sow/year ranges from 1.1 to 1.5. However, some native pig
breeds have a higher reproductive performance , for example Mong
Cai pig (10 - 15 piglets/litter and 1.5-2 litters/year) (Phan Xuan Hao
and Thanh Ngoc Van, 2010; Tu Quang Hien et al., 2004; Nguyen
Manh Cuong et al., 2010; Ho Trung Thong et al., 2011), Lung Pu pig
(Vu Ngoc Son et al., 2009; Nguyen Van Duc et al., 2008; Trinh
Quang Phong et al., 2011; Trinh Quang Phong et al., 2012; Nguyen
Van Duc et al., 2010) and other pig breeds (Vu Dinh Ton and Phan
Dang Thang, 2009; Vu Dinh Ton et al., 2012; Quach Van Thong,
2009).

1.3. Growth, productivity and quality of the meat of
native pigs
In recent years, there have been many works dealing with
growth, meat production and quality of meat of native pig breeds
such as Nguyen Ngoc Phuc, et al., 2010; Phan Xuan Hao and Ngoc
Van Thanh, 2009; Tran Thanh Van and Dinh Thu Ha, 2005; Le Dinh
Cuong et al., 2004; Ho Trung Thong et al., 2011. These works have
been concluding that growth, meat production of native pig breeds
are not high.
There have also been some studies and publications on growth,
meat production of Lung Pu pigs (Vu Ngoc Son et al., 2009; Nguyen
Van Duc et al., 2008; Trinh Quang Phong, 2011; Nguyen Van Duc et
al., 2008) and of Ban pigs (Vu Dinh Ton and Phan Dang Thang,
2009; Quach Van Thong, 2009). However, these studies have not
been providing us with a sufficient information on growth and meat
production of these native pigs. Pig meat quality has nor been
studying in the above studies.
Therefore, in this study, in addition to assessing reproduction,
growth, meat production and meat quality of these pigs will be
systematically and fully evaluated in both sexes.


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1.4. Genes related to pig meat quality
In Vietnam, up to now there have been some studies to identify
molecular markers (DNA) in animals using molecular genetic
techniques to find out DNA markers associated with production traits
(Le Minh Sat, 1997, Dinh Van Chinh et al., 1999; Nguyen Ngoc
Tuan and Tran Thi Dan, 2005; Do Vo Anh Khoa et al., 2010).
A few studies on genes related to meat quality and productivity

of indigenous breeds and hybrid breeds (domestic x exotic breeds)
have been published in the country (Nguyen Van Hau et al., 2000; Le
Thi Thuy et al., 2004; Nguyen Van Cuong et al., 2003).
PRKAG3 gene has been considering to have possible effects on
meat quality of many exotic breeds (Ciobanu et al., 2001; Milan et
al., 2000; Anna Grandlund et al., 2011; Meadus et al., 2002 ; Skrlep
et al., 2009) and a number of Chinese indigenous pig breeds (LuSheng Huang et al., 2004). However, this gene has not been studying
in any pig breed in Vietnam. Therefore, polymorphism of PRKAG3
gene in two pig breeds of Lung Pu and Ban and influences of this
genetic polymorphism on meat quality are questions to be answered
in this study.
CHAPTER II
MATERIALS AND METHODS
2.1. Research subjects
- Evaluate the performance characteristics of 303 Lung Pu pigs,
277 Ban pigs in 2 generations: parents and offspring. Assessment of
productivity of 218 litters in Lung Pu pig, 213 litters in Ban pig.
- Assess the growth (37 Lung Pu pigs, 38 Ban pigs), the ability
for the meat producing and meat quality (30 Lung Pu pigs, 29 Ban
pigs).
- Identify genetic polymorphism of PRKAG3 in 150 individual
Lung Pu pigs and 150 individual Ban pigs.
- Lung Lung Pigs were raised in Lung Pu commune, Meo Vac
district, Ha Giang province. Ban pigs are raised in Phu Cuong
commune, Tan Lac district, Hoa Binh province.
- Study was carried out from 01/2012 to 06/2015.


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2.2. Research content

2.2.1. Identifying appearance characteristics and performance of
Lung Pu and Ban pig
- Characteristics of hair coat and skin color of Lung Pu and Ban
pig.
- Reproductive performnce of Lung Pu and Ban pig.
- Growth of Lung Pu and Ban pig.
- Carcass yield and meat quality of Lung Pu and Ban pigs.
2.2.2. Identifying genetic polymorphisms of PRKAG3 gene in Lung
Pu and Ban pigs and effects of this gene on meat quality of Ban pig.
- Identify PRKAG3 gene polymorphism in Lung Pu and Ban
pig.
- Determine possible effects of PRKAG3 gene polymorphisms
on meat quality of Ban pig.
2.3. Methods
2.3.1. Identifying appearance characteristics and performance of
Lung Pu and Ban pig
2.3.1.1. Appearance characteristic of Lung Pu and Ban pigs
Study on appearance characteristics mainly focused on evaluation of
hair coat and skin color characteristics of Lung Pu and Ban pig
according to criteria: Lung Pu pigs: (i) black body, (ii) black body
with 6 white-dotted points (from four elbows to the feet, forehead
and tip of the tail) and (iii) black body with white rippling abdomen;
Ban pigs: (i) black body, (ii) black body with 4 white pedicures and
(iii) body black with a white rippling abdominal.
2.3.1.2. Reproductive performance of Lung Pu and Ban pigs
- For each breed, 40 gilts and replaced boars were chosen.
Selected individual boar, gilt selected was not relative. Average
weight of selected pigs was 20 kg/head.
- Two experiments for two breeds were conducted. Each
experiment was arranged in 10 households (4 pigs/household) with

similar farming conditions. 40 sows were put in pair with 4 males for
rotational mating from first litter to the sixth. Vietnam standards ISO
1547-2007 for nutritive requirements was used for rearing animals.


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- Monitoring performance of experimental pigs at farms with
recording books.
2.3.1.3. Growth of Lung Pu and Ban pigs
- Before observation, experimental pigs were chosen and
identified by ear tags, weighed. The experiment was repeated 2
times, and ratio of castrated male to female was 1 to 1.
- Raising condition: Locally available feeds were used to
formulate rations according Vietnam standards ISO 1547-2007 for
nutritive requirements of native pigs.
- Monitoring growth of pigs from 2 months of age to 8 month
old by monthly and individually weighing and recording.
2.3.1.4. - Carcass yield and meat quality of Lung Pu and Ban pigs.
a) Yield of meat:
- Choosing pigs for slaughtering: Slaughtered pigs were selected
from pigs in growth experiments based on their growth rate. Ratio of
male to female was 1 to 1.
- The classical method was used for slaughtering tests, the left
half of carcarss was used to separate meat, fat, bone, skin and
weighed different parts according to Vietnam Standard (TCVN 38991984).
b) Quality of meat:
- Samples of fillet meat were taken immediately after
slaughtering in position of 13th rib to and 14th rib, preserved in ice
boxes and transported to the laboratory. Fillet meat was cut into 3
parts with a thickness of 3 cm, then stored at 4°C for analysis of meat

quality.
- pH45 and pH24 values, meat color, water loss rate of
preservation and processing were determined by a method of Warner
et al. (1997).Toughness (softness) of meat was determined by
method of Channon et al. (2003).
2.3.2. Identifying genetic polymorphisms of PRKAG3 gene in Lung
Pu and Ban pigs and effects of this gene on meat quality of Ban pig.
2.3.2.1. Identifying genetic polymorphisms and sequencing of
PRKAG3 gene


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- Extraction of DNA from tissue samples was done by using
DNA kit of Bioneer (South Korea).
- Identify PRKAG3 polymorphic gene segments was performed
using PCR-RFLP:
- Sequencing PRKAG3 gene was done by using Sanger method
on sequencer machine ABI3130.
- Analysis of PRKAG3 gene sequences was performed with a
profestional software BioEdit v7.2.5 at Key Laboratory Animal
Cells, NIAS.
2.3.2.2. Determine possible effects of PRKAG3 gene polymorphisms
on meat quality of Ban pig.
After identifying PRKAG3 genotype polymorphism, individual
pigs carrying the typical genotype were marked, continuously
monitored up to 8 months of age and slaughtered to evaluate the
relationship of each genotype with meat quality.
2.4. DATA PROCESSING
Data were statistically analyzed using SAS 9.0 software (2002),
to calculate the statistical parameters LSM, SE, LSMeans with pair

comparison pdiff Tukey corrected using Turkey method with
analytical model as follows:
Yijkl = μ + Bi + Lj + Nk + Ɛijkl
Where: Yijkl: Obsevered values of parameter
μ: Population average
Bi: Effect of boar i (Lung Pu = 1 to 4; iBAN = 1 to 4).
Lj: Effect of parity j (j = 1 from 1 to 6).
Nk: Effect of the year k (k = from 2012 to 2015).
Ɛijkl: random error.

CHAPTER III
RESULTS AND DISCUSSION
3.1. APPEARANCE CHARACTERISTICS AND PERFORMANCE
OF LUNG PU AND BAN PIGS

3.1.1. Appearance characteristic of Lung Pu and Ban pigs
Lung Pu pig had prominent features such as a black body with 6
white-dotted points. The percentage of pig having this character was


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relatively high (65.68%). This figure for parents' generation was
higher than that of the offspring generation (75.00 vs. 64.09%) and
the difference was not statistically significant (P <0.05).
Ban pig had typical features such as a fully black body with 4
white toe nails (59.21%), 65.91% of parents generation had this
character whereas the figure for offsping generations was 57.94%.
There was not any statistical difference between generations (p =
0.3239).
Table 1: Some appearance characteristics of Lung Pu and Ban pigs

Total

Features
n
Lung Pu pigs
Full body black
Full body black with 6
white points (4 elbows to
nails, forehead and tip of
the tail)
Full body black with a
white rippling abdominal
Total
Ban pigs
Full body black
Full body black with 4
white pedicures
Full body black with a
white rippling abdominal
Total

54

%

Parental
generation
n
%

17.82 5

Progeny
n

%

X2
(p
value)

11.36 49 18.92 0.2260

199 65.68 33 75.00 166 64.09 0.1589

50
303
62

16.50 6
100

44

22.38 8

13.64 44 16.99 0.5797
100

259 100

18.18 54 23.18 0.4660

164 59.21 29 65.91 135 57.94 0.3239
51
277

18.41 7
100

44

15.91 44 18.88 0.6405
100

233 100

3.1.2. Reproductive performance of Lung Pu and Ban sows
3.1.2.1. Factors affecting the reproductive performance of sows
Reproductive performance of Lung Pu was influenced by parity
(P <0.05 to P <0.001). A strongest factors were the average birth
weight/litter and average weaning weight/litter (P <0.001). The
average weaning weight/piglet was not affected by the parity. The
father (boar) and the year did not affect the reproductive performance
of Lung Pu pig.


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Table 2: Factors affecting the reproductive performance of Lung Pu sows
Parameters

Number born alive/litter
(piglets)
Number of pigs weaned
/litter (piglets)
Total birth weight/litter (kg)
Birth weight/piglet (kg)
Weaning weight
/litter (kg)
Weaning weight
/piglet (kg)
Number of parity/sow/year
(litter)

Boars
ns

Parity
**

Years
ns

R2
0.1886

ns

**

ns

0.2013

ns
ns
ns

***
*
***

ns
ns
ns

0.2563
0.1061
0.1909

ns

ns

ns

0.0909

ns

*

ns

0.093
5

Note: NS: P≥0,05; *: P<0,05; ** : P<0,01 ; ***: P<0,001

All parameters of reproductive performance of Ban pig were
affected by litter number from P < 0.05 to P < 0.001, except for
weaning weight/piglet. In which, the number of newborn piglet
alive/litter, number weaning piglet/litter, birth weight/litter and
weaning weight/litter were strongly influenced (P <0.001) and
weaning weight/piglet was not affected by parity. Number of
parity/sow/year was influenced by parity and year (P <0.05).
Table 3: Factors affecting the reproductive performance of Ban pig
Parameters
Number born alive/litter (piglets)
Number of pigs weaned
/litter (piglets)
Total birth weight/litter (kg)
Birth weight/piglet (kg)
Weaning weight
/litter (kg)
Weaning weight
/piglet (kg)
Number of parity/sow/year (litter)

Boars
ns

Parity
***

Years
ns

R2
0.2471

ns

***

ns

0.2690

ns
ns

***
*

ns
ns

ns

***

ns

ns

ns

ns

ns

*

*

0.2994
0.1778
0.239
3
0.042
4
0.0996

Note: NS: P≥0,05; *: P<0,05; ** : P<0,01 ; ***: P<0,001

3.1.2.2. Reproductive performance
a) The physiological and reproductive parameters


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Age at first estrus of Lung Pu pig was 197.18 days when their
body weight was about 28.44 kg. Figures for Ban pig were 201.78
days and 22.46 kg, respectively. Ban pigs went on the first estrus
later than that of Lung Pu and their body weight at first estrus was
lower than that of Lung Pu. Therefore, it seemed that that there was
a big difference in age at the first estrus and their body weight at the
first estrus between two indigenous breeds of pigs. Age at the first
mating of Lung Pu pig was 250.20 days when their body weight was
around 39.08 kg. However, the figures for Ban pig were 250.38 days
and 31.37 kg respectively.
Table 4: Some physiological and reproductive parameters of Lung Pu
and Ban pigs
Lung Pu pigs
Ban pigs
Paremeters
n
LSM
SE
n
LSM
SE
Age at the first
estrus
40
197.18 0.76
40
201.78
2.69
(days)
Age at the first

insemination
40
250.20 1.78
40
250.38
2.06
(days)
Weight of the first
40
28.44 0.15
40
22.46
0.42
estrus (kg)
Weight of the first
40
39.08 0.40
40
31.37
0.42
insemination (kg)

b) Phenotypic value of reproductive performance
Age at the first farrowing of Lung Pu and Ban pigs was 363.75
and 363.70 days, respectively. Vu Ngoc Son et al. (2009) reported
that this figure of Lung Pu pig was 385.30 days; According to Vu
Dinh Ton and Phan Dang Thang (2009), Quach Van Thong (2009),
age at the first farrowing of Ban pig was 388.96 days and 394.80
days respectively. It meant that results of our study were lower for
both breeds.

Number of newborn piglet/litter for Lung Pu pig was 7.59,
compared to 5.87 as reported by Vu Ngoc Son et al. (2012). For Ban
pig, the newborn piglet/litter was 7.31, in agrrement with 7.33 as
reported by Vu Dinh Ton, Phan Dang Thang,2009) .


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Number of newborns alvie/litter for Lung Pu and Ban pig was
7.42 and 7.10 respectively. These figures were in line with results
reported by Trinh Quang Phong and Dao Duc Tha (2012) who
studied on Lung Pu pig. However, the figures were higher than those
in Ban pig reported by Vu Dinh Ton and Phan Dang Thang (2009),
(2012).
Table 5: Reproductive performance of Lung Pu and Ban sows
Parameters
Age at the first
farrowing (day)
Total number of pigs born/litter
(piglets)
Number born alive/litter (piglet)
Birth weight/piglet (kg)
Total birth weight/litter (kg)
Number of pigs kept
/litter (piglets)
Time of weaning (days)
Number of weaned pigs/litter
(pigs)
Survival rate to weaning (%)
Weaning weight/pig (kg)
Total weaning weight/litter (kg)

Time insemination pregnancy
after weaning (day)
The gestation period (days)
The distance between the two
parities (days)
Number of parity/sow/year
(litter)

n

Lung Pu Pig
LSM
SE

n

Ban Pig
LSM SE

40

363.75 1.78

40

363.70 2.05

218

7.59

0.09 213

7.31

0.11

218
83
218

7.42
0.49
3.63

0.09 213
0.00 84
0.05 213

7.10
0.44
3.10

0.10
0.01
0.05

218

7.23

0.08 213

6.97

0.09

218

45.17

0.10 213

45.49

0.12

218

7.03

0.08 213

6.76

0.09

218
82
218

97.05
4.17
29.03

0.41 213
0.02 81
0.36 213

97.25
3.80
25.31

0.39
0.05
0.38

178

27.33

0.95 173

19.37

0.35

218

114.33 0.07 213 114.38 0.08

178

186.72 0.99 173 179.38 0.38

178

1.96

0.01 173

2.04

0.00

Newborn piglet weight/piglet of Lung Pu and Ban pig was 0.49
kg and 0.44 kg, respectively. Newborn piglet weight/litter of Lung Pu
(3.63 kg) was higher than that of Ban pig (3.10 kg).
Number weaned pig/litter of Lung Pu pig was 7.03 pigs, higher
than the result published (6.12 pigs) of Trinh Quang Phong and Dao
Duc Tha, 2012, but lower than that reported by Nguyen Van Duc


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(2013) at Meo Vac (7.12 pigs) and Vi Xuyen (7.16 pigs). Number
weaned pigs/litter of Ban pig was 6.76 pigs, higher than results in
Ban pigs reported by Quach Van Thong (2009), Vu Dinh Ton and
Phan Dang Thang (2009), Vu Dinh Ton et al. (2012) (5.95; 5.80 and
5.63 weaned pigs/litter).
At 45.17 days of weaned age, the weight of Lung Pu pig was

4.17 kg and weaning weight/litter was 29.03 kg. Ban pigs were
weaned at 45.49 days, weaned weight was 3.80 kg/pig and weaned
weight/litter was 25.31 kg.
Farrowing interval of Lung Pu and Ban pigs was 186.72 and
179.38 days, respectively. Number farrowing/sow/year of Lung Pu
and Ban pigs was rather high (1.96 and 2.04 farrowing/sow/year).
- Reproductive performance of Lung Pu sows by parity
Reproductive performance of Lung Pu (Figure 1) from parity 1
to parity 6 showed that the number newborn piglet/litter was lowest
at parity 1, increased rapidly at parity 2, reached the highest at parity
4 and began to decline at parity 5 and 6. Similarly, the number
newborn alive/litter was lowest at parity 1 (6.61 piglets/litter). There
was a difference in the number newborn alive/litter among parity 1,
2, 3 and 4 (P<0.05). The number newborn alive/litter was the highest
(7.96 piglets/litter) at parity 4.
Newborn weight/piglet at parity of 1, 2, 5 and 6 was statically
different from newborn weight/piglet at parity 3 and 4 (P<0.05).
Newborn weight/litter was also increased as the number newborn
alive/litter and there was a difference in Newborn weight/litter
among parity 1, 5 and 6 with parity 2, 3 and 4 (P<0.05).
The number weaned pig/litter was the lowest in parity 1 (6.48
pigs) and statistically differed (P<0.05) with the number weaned
pig/litter at parity of 2, 3, 4. However, no difference was found in the
number weaned pig/litter between parity 5 and 6. Weaned pig
weight/head was different between the parity 5 and 6, with parity 2, 3
and 4 (P<0.05) and weaned pig weight/litter peaked at parity 3, was
the lowest in the parity 6 and was a difference among parities of 1, 5,
6 from parities of 2, 3, 4.

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Figure 1: Number newborn piglet alive/litter and number weaned
pig/litter of Lung Pu pig by parity

- Reproductive performance of Ban sow by parity:
The number and weight of Ban pig (Figure 2) had a tendency:
increased from parity 1, peaked at parity 4 and decreased in parity 5
and 6. The number newborn piglet/litter (6.51 piglet) and the number
newborn alive/litter (6.19) in parity 1 was low and statistically
significant different (P<0.05) from parities 2 to 4, but not different
from parities 5 and 6. The newborn piglets/litter and number
newborn alive/litter were the highest in parity 4 (8.40 and 8.15).

Figure 2: The number newborn piglet/litter (6.51 piglet) and the
number newborn alive of Ban pig by parity.

Newborn piglet weight/litter at parity 1 was different from parity
3, 4, 5, 6. Newborn piglet weight/litter at parity 2 was significantly
different from parity 3, 4, 5 (P<0.05). Newborn piglet weight/litter


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was the lowest at parity 1 (2.41 kg) and significantly different from
parities 2, 3, 4, 5 (P<0.05).
Number weaned pigs/litter was the lowest in parity 1 (5.83
piglets/litter) and statistically significant different (P<0.05) with
parity from 2 to 4. The weaned pig weight /pig among parities were
not different (P>0.05). However, weaned pig weight/litter at parity 1
was different from parity from 2 to 4 (P<0.05).

Figure 1 and in Table 2 showed that reproductive performance
of Ban and Lung Pu such as the number of pig, body weight varied
and increased from parity 1 to parity 2, peaked at parity 3 and parity
4, and started to decrease at parity 5 and 6.
3.1.3. Growth, meat production and meat quality of Lung Pu and
Ban pigs
3.1.3.1. Body weight and body weight gain of Lung Pu and Ban pig
over months of age
a) Body weight of Lung Pu over months of age
Body weight of Lung Pu pig at 2 months of age was 6.99 kg,
differences in body weight of castrated boars and gilts were not
statistically significant (P>0.05) (7.13 vs. 6.87 kg)
Table 5: Body weight of Lung Pu pigs over the months of age

Months
of age
2
3
4
5
6
7
8

n
37
37
37
37
37

37
37

Average
LSM SE
6.99 0.08
11.52 0.10
17.18 0.16
23.89 0.17
31.07 0.22
38.17 0.25
44.80 0.27

Castrated boars
n LSM SE
18 7.13 0.12
18 11.75a 0.15
18 17.49a 0.24
18 24.35a 0.25
18 31.77a 0.33
18 39.04a 0.36
18 45.83a 0.39

n
19
19
19
19
19
19

19

Gilts
LSM
6.87
11.29b
16.87b
23.43b
30.36b
37.31b
43.77b

SE
0.11
0.13
0.22
0.23
0.30
0.34
0.36

Note: means in the same rows bearing different superscripts were statistically
different (P<0.05)

At 8 months of age, on average, body weight of Lung Pu pig
was 44.80 kg, weight of castrated boars (45.83 kg) was higher than
that of gilts (43.77 kg) and difference in weight of castrated boars
and gilts was statistically significant (P<0.05). Body weight of

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castrated boars at ages of 3 to 8 months was statistically significant
different (P<0.05) with body weight of gilts at similar ages.
Table 6: Body weight of Ban pigs over the months of age

Months
of age n
2
38
3
38
4
38
5
38
6
38
7
38
8
38

Average
LSM SE
5.49 0.11
8.15 0.09
11.53 0.12
15.78 0.13
20.73 0.17
26.26 0.23

31.85 0.24

Castrated boars
n LSM SE
18 5.63 0.17
18 8.35a 0.14
18 11.69 0.17
18 16.08a 0.19
18 20.98 0.25
18 26.72 0.34
18 32.54a 0.34

n
20
20
20
20
20
20
20

Gilts
LSM
5.36
7.94b
11.37
15.48b
20.47
25.81
31.16b

SE
0.16
0.13
0.16
0.18
0.23
0.32
0.32

Note: means in the same rows bearing different superscripts were statistically
different (P<0.05).

The body weight of Ban pig at 2 months age was 5.49 kg and
that at 8 months age was 31.85 kg. Average body weight of Ban pig
at 2, 3, 4, 5, 6 and 7 months age was 8.15; 11.53; 15.78; 20.73 and
26.26 kg, respectively. Results in table 7 also showed that the body
weight of between Ban castrated boars and gilts at the age of 3, 5 and
8 months was statistically significant different (P<0.05), but the
difference at 4, 6 and 7 months age was not statistically significant
(P>0.05).
b) Body weight gain of Lung Pu and Ban pig over months of age
Results in Table 8 showed that body weight gain of Lung Pu
pigs was 150.71; 188.50; 223.73; 239.23; 236.93; 220.80 and 209.98
g/day for periods of time from 2 to 8 months age, respectively.
Body weight gain of castrated boars was alway higher than that
of gilts (P<0.05).
Body weight gain of Lung Pu rapidly increased from 2 to 3
months and 6-7 months of age and began to decrease in from 6-7
months of age.

Results in Table 9 showed that body weight gain of Ban pigs
was 88.45; 112.82; 141.56; 165.01; 184.56 and 186.20g/day, for
periods of time from 2 to 8 months age, respectively.


16
Table 7: Body weight gain of Lung Pu pig over the months of age
Months
of age
2-3
3-4
4-5
5-6
6-7
7-8
2-8

n
37
37
37
37
37
37
37

Average
LSM
150.71
188.50

223.73
239.23
236.93
220.80
209.98

SE
3.60
2.85
4.27
4.37
4.02
5.53
1.62

Castrated boars
n
LSM
SE
18 153.92 5.30
18 191.18 4.19
18 228.63 6.28
18 247.45 6.42
18 242.35 5.92
18 226.27 8.14
18 214.97a 2.38

n
19
19

19
19
19
19
19

Gilts
LSM
147.50
185.83
218.83
231.00
231.50
215.33
205.00b

SE
4.88
3.87
5.79
5.92
5.45
7.50
2.20

Note: means LMS in the same rows bearing different superscripts were statistically different
(P<0.05).

Body weight gain of Ban pigs for the entire period (2-8 months)
was 146.44g/day. Body weight gain of castrated boars was higher

than that of gilts, but difference in body weight gain between
castrated boars and gilts, was not statistically significant (P>0.05).
(P<0.05). However, difference in body weight gain between castrated
boars and gilts for the whole period from 2 to 8 months of age was
significantly different (P<0.05).
Table 8: Body weight gain of Ban pig over the months of age

Months
of age
2-3
3-4
4-5
5-6
6-7
7-8
2-8

n
38
38
38
38
38
38
38

Average
LSM SE
88.45 2.18
112.82 1.73

141.56 3.80
165.01 3.52
184.56 5.19
186.20 5.65
146.44 1.34

Castrated boars
n LSM SE
18 90.74 3.16
18 111.48 2.51
18 146.11 5.51
18 163.52 5.11
18 191.30 7.53
18 194.07 8.20
18 149.54a 1.95

n
20
20
20
20
20
20
20

Gilts
LSM
86.17
114.17
137.00

166.50
177.83
178.33
143.33b

SE
3.00
2.38
5.22
4.85
7.14
7.78
1.85

Note: means LMS in the same rows bearing different superscripts were statistically
different (P<0.05)

3.1.3.2. Carcass yield of Lung Pu and Ban pig
Slaughtered results of Lung Pu carcase yield was presented in
Table 10. Slaughtered weight of Lung Pu at 8 months of age, hoof
percentge, dressing percentage and lean meat percentage were 44.71
kg, 74.28%, 62.16%; and 38.80%, respectively. Lean percentage of
castrated boars was not statistically significant different (P>0.05)
with that of gilts (P>0.05).


17
Table 9: Carcass yield of Lung Pu pig
Parameters
n

Carcass weight (kg)
30
Hoof percentage (%)
30
Dressing percentage (%) 30
Lean meat percentage (%) 30
Fat percentage (%)
30
Bone percentage (%)
30
Skin percentage (%)
30
Back fat thickness (mm) 30

Average
LSM SE
44.71 0.31
74.28 0.30
62.16 0.21
38.80 0.20
29.55 0.30
19.22 0.37
12.43 0.20
28.74 0.57

Castrated boars
n LSM SE
15 45.67a 0.44
15 74.88 0.42
15 62.97a 0.29

15 38.52 0.28
15 29.84 0.43
15 19.32 0.53
15 12.33 0.28
15 29.31 0.80

n
15
15
15
15
15
15
15
15

Gilts
LSM
43.75b
73.68
61.35b
39.08
29.26
19.13
12.54
28.17

SE
0.44
0.42

0.29
0.28
0.43
0.53
0.28
0.80

Note: means LMS in the same rows bearing different superscripts were statistically different
(P<0.05).

Slaughtered results of Ban carcase yield was presented in Table
11. Slaughtered weight of Ban at 8 months of age, hoof percentge,
dressing percentage and lean meat percentage were 32.14 kg,
71.04%, 62.94% and 40.31%; respectively. Lean percentage of Lung
Pu pigs was rather high. Lean percentage of castrated boars was
statistically significant different (P>0.05) with that of gilts (P>0.05).
Table 10: Carcass yield of Ban pig
Parameters
n
Carcass weight (kg)
29
Hoof percentage (%)
29
Dressing percentage (%) 29
Lean meat percentage (%) 29
Fat percentage (%)
29
Bone percentage (%)
29
Skin percentage (%)

29
Back fat thickness (mm) 29

Average
LSM SE
32.14 0.26
71.04 0.22
62.94 0.34
40.31 0.18
23.58 0.42
21.09 0.35
15.02 0.37
26.18 0.34

Castrated boars
n LSM SE
14 32.71a 0.37
14 71.50 0.32
14 63.09 0.49
14 39.74b 0.26
14 23.83 0.60
14 21.12 0.50
14 15.30 0.54
14 26.28 0.49

n
15
15
15
15

15
15
15
15

Gilts
LSM
31.57b
70.58
62.80
40.88a
23.33
21.05
14.74
26.09

SE
0.35
0.31
0.47
0.25
0.58
0.49
0.52
0.48

Note: means LMS in the same rows bearing different superscripts were statistically different
(P<0.05).

3.1.3.3. Meat quality of Lung Pu and Ban pig

Results in Table 12 showed that the pH45 of loin meat of Lung
Pu castrated boars (6.29) and gilts (6.15) was significantly. At 24
hours post-mortem, pH24 value measured in loin meat of Lung Pu
was reduced to 5.60. There was a significant difference in pH24h
measured in loin meat between castrated boars and gilts (P<0.05).


18
Table 12: Meat quality of Lung Pu pig
Parameters

n
30
30
30
30
30

Average
LSM SE
6.22 0.02
5.60 0.02
53.33 0.33
15.57 0.16
9.00 0.24

Castrated boars
n LSM SE
15 6.29a 0.02
15 5.55b 0.03

15 53.09 0.47
15 15.20b 0.22
15 8.88 0.33

n
15
15
15
15
15

Gilts
LSM
6.15b
5.66a
53.57
15.95a
9.11

SE
pH45
0.02
pH24h
0.03
L* (lightness)
0.47
a* (Red)
0.22
b* (Yellow)
0.33

Water loosing percentage
b
a
30 1.96 0.10 15 1.68 0.14 15 2.24 0.14
by preservation (%)
Water loosing percentage
30 30.40 0.54 15 30.95 0.76 15 29.86 0.76
by processing (%)
Toughness (N)
30 60.66 1.22 15 62.19 1.72 15 59.12 1.72
Note: means LMS in the same rows bearing different superscripts were statistically different
(P<0.05).

pH45 and pH24 values of Ban pigs were presented in Table 13,
pH45 value of castrated boars was 6.18, and gilts was 6.20 and no
significant differences was found (P>0.05). After 24 hours of
preservation, average pH24 values decreased to 5.69 and these values
were not different (P>0.05) between castrated boars and gilts.
Table 13: Meat quality of Ban pig
Parameters
pH45
pH24h
L* (lightness)
a* (Red)
b* (Yellow)
Water loss
percentage by
preservation (%)
Water loss
percentage by

processing (%)
Toughness (N)

Average
n LSM SE
29 6.19 0.03
29 5.69 0.02
29 55.36 0.53
29 14.96 0.38
29 8.42 0.18

Castrated boars
n LSM SE
15 6.18 0.04
15 5.67 0.03
15 55.61 0.73
15 14.90 0.53
15 8.55 0.25

n
14
14
14
14
14

29

15 2.03b

0.16

14 2.53a 0.16

29 28.94 0.39

15 28.06a 0.54

14 29.82b 0.56

29 61.48 3.70

15 65.04 5.15

14 57.91 5.33

2.28

0.11

Gilts
LSM
6.20
5.71
55.11
15.02
8.28

SE
0.04

0.04
0.76
0.55
0.25

Note: means LMS in the same rows bearing different superscripts were statistically different
(P<0.05).


19
Research results on Lung Pu (table 12) and Ban pig (table 13)
showed that the color of Lung Pu and Ban meat was within an
acceptable limits. These parameters in each breed were not
statistically significant different (P>0.05) between castrated boars
and gilts, except for a* of Lung Pu pig (P<0.05).
The water loss percentage by preservation of Lung Pu pig was
1.96% and this difference was statistically significant (P<0.05)
between castrated boars and gilts. The water loss percentage by by
processing was 30.40% and there were no differences between
castrated boars and gilts (P>0.05).
The water loss percentage by preservation of Ban pig was
2.28%, average water loss percentage by processing was 28.94%,
The water loss percentage by preservation and processing of gilts
were higher than that of castrated boars (P<0.05).
The toughness of Lung Pu pigs (60.66 N) was lower than Ban
(61.48 N). It meant that meat of Lung Pu pig was more tender than
that of Ban pigs
3.2. PRKAG3 gene polymorphism on Lung Pu, Ban pigs and Its
effects on meat quality of Ban pig
3.2.1. PRKAG3 Gene polymorphism

3.2.1.1. Specific amplification of exon 1 and exon 3 region on
PRKAG3 gene
Amplication of PRKAG3 gene fragment on exon 1 and exon 3
was successfully with sizes of 270 bp và 258 bp. It was fuond out that
only one light DNA band appeared. The PRKAG3-exon1 and exon 3
gene fragments were multiplied corresponding to gene coding
sequence
with
codes
PRKAG3
AF214521.1
on
www.ncbi.nlm.nih.gov gene banks.
3.2.1.2. Exon 1 and exon 3 regional polymorphic of PRKAG3 gene
a) The exon 1 polymorphism of PRKAG3 gene (G52S/HphI and
T30N/StyI)
The first polymorphisms at positions 154, which had changed
nucleotide between G and A (G154A) and led to change G and S
amino acid at position 52 in the polypeptide chain (G52S). The
second polymorphism at position 89, which had changed between


20
nucleotides C and A (C89A) and led to change amino acid at position
30 (T30N).
At position of G52S polymorphism, we discovered 2 types of S
and G alleles, corresponding to 3 SS, SG and GG genotypes. At
position of T30N, 2 types of allele T and N were detected
corresponding to 3 genotypes of NN, TN and TT and appeared in two
pig breeds studied.

Polymorphism at position G52S and T30N of PRKAG3 gene in
Lung Pu and Ban pigs was in agreement with the work done by
Sheng Huang Lu et al. (2004), who found 9/21 indigenous breeds in
China having polymorphism at position G52S and T30N of PRKAG3
gene .
b) PRKAG3 exon 3 gene Polymorphism (V199I/BsaHI and
R200Q/BsrBI)
Analyzing the V199I exon 3 gene PRKAG3 polymorphism in
Lung Pu and Ban pigs, we detected 2 alleles that were V and I
(Figure 15). However, only 2 genotypes of VV and IV were found.
The above results were corresponding to Lu-Sheng Huang et al.
(2004) who reported that I allele was not present in Chinese
indigenous breeds and occured with a very low frequencies in a
number of exotic breeds.
At R200Q position in Ban and Lung Pu pigs, we only
discovered R allele corresponding to RR genotype.
From analysis of results, it was shown that primers used to
specifically amplify and analyze the polymorphism in PRKAG3 3
gene exon 1 and exon 3 regions in Lung Pu and Ban pigs were
specific, accurate and consistent with the study published in the
world.

Figure 1: Image of PRKAG3 gene
sequencing exon 1 region

Figure 2: Image of PRKAG3 gene
sequencing exon 3 region


21

3.2.1.3. Sequencing
Results of PRKAG3 exon 1 gene sequence and analysis showed
that these gene regions were amplified specifically, polymorphic
points identified at A89C A154G positions were completely accurate
and corresponded to points cut by HphI and StyI enzymes (Figure 3).
The results of sequence and analyze PRKAG3 exon 3 gene
region also showed that this gene segment was multiplied specifically
and exactly. These polymorphisms determined were in accordance
with the cutting point of BsaHI and BsrBI enzyme as reported
(Figure 4).
3.2.2. Genotype and allele frequencies of PRKAG3 gene
polymorphisms
3.2.2.1. Lung Pu pigs
Results of allele and genotype frequency analysis in the exon 1
and PRKAG3 exon 3 polymorphisms of Lung Pu pigs were presented
in Table 14.
Table 11: Allele and genotype frequency in PRKAG3 gene
polymorphisms of Lung Pu pigs
Polymorphism
Genotype frequency
Allele frequency
G52S/HphI (n=150)
GG
SG
SS
G
S
1.00
0
0

1.00
0.00
T30N/StyI (n=150)
NN
TN
TT
T
N
0
0
1
1.00
0.00
V199I/BsaHI (n=150) VV
IV
II
V
I
1
0
0
1.00
0.00
R200Q/BsrBI (n=150) RR
QR
QQ
R
Q
1
0

0
1.00
0.00

G52S/HphI polymorphism only appeared G allele,
polymorphism T30N/StyI appeared T allele, V199I/BsaHI
polymorphisms appeared V allele and R200Q/BrsBI polymorphism
only appeared R allele. Therefore, at each polymorphism, both
genotypes and alleles frequency was 1.
Research results indicated that PRKAG3 gene polymorphisms
was not appeared at 4 polymorphism position in the Lung Pu pig
population.


22
3.2.2.2. Ban pig
a) G52S/HphI polymorphism
Evaluation of G52S polymorphism in Ban pig population
showed that the frequency of homozygous genotype GG (0.82) was
higher than the frequency of heterozygous genotype SG (0.11) and
SS (0.07). G allele frequency (0.88) was higher than that of S allele
frequency (0.12).
Table 12: Allele and genotype frequency in PRKAG3 gene
polymorphisms of Lung Pu pigs
Polymorphism
Genotype frequency
Allele frequency
G52S/HphI
GG SG
SS

G
S
(n=150)
0.82 0.11
0.07
0.88
0.12
NN TN
TT
T
N
T30N/StyI (n=150)
0.01 0.03
0.96
0.97
0.03
IV
II
V
I
V199I/BsaHI (n=150) VV
0.98 0.02
0.00
0.99
0.01
QQ
R
Q
R200Q/BsrBI (n=150) RR QR
1.00 0.00

0.00
1.00
0.00

Results of T30N/StyI polymorphism analysis in Ban pigs
showed that T allele frequency (0.97) was higher than that of N allele
(0.03). TT genotype frequency was the highest (0.96) while the 2
remaining genotypic frequencies was very low in the population (TN
= 0.03, NN = 0.01).
At V199I/BsaHI polymorphism position, the frequency of V
allele was high (0.99), while the frequency of I allele was very low in
the population (only 0.01). In this study, only two IV and VV
genotypes were found.
R200Q/BsrBI polymorphism showed frequencies of RR
homozygous genotype, R allele frequency were 1.00 and 1.00,
respectively.
Thus, corresponding genotypes existed in each polymorphism of
Ban pigs in the study, specifically G52S/HphI and T30N/StyI
polymorphism had 3 genotypes in the population; V199I/BsaHI
polymorphisms, VV genotype was more dominant in the population
than IV genotype, yet R200Q/BsrBI polymorphism. II genotype have
not been found and only RR genotype appeared.


23
3.2.3. Influence of PRKAG3 gen polymorphism on meat quality
of Ban pigs
3.2.3.1. G52S/HphI polymorphism
Water loss percentage by processing of SG genotype was higher
than that of GG genotype. Water loss percentage by preservation of

SG genotype was lower than that of GG genotype. However, no
difference in water loss percentage between two genotypes was
found (P>0, 05).
L* values of GG genotype were higher than that of SG
genotype, and differences were statistical significant (P<0.05).
a* value of GG genotype was lower than SG genotype, while b*
value of GG genotype was also lower than that of SG genotype.
However, the difference was not statistically significant between the
two genotypes (P > 0.05).
The toughness of Ban pig in animals with GG genotype (61.89
N) was lower than that in animals with SG genotype (66.10 N).
However, this difference was not statistically significant (P>0.05).
Table 13: Influence of G52S/HphI polymorphism on meat quality of Ban
pigs
Parameters
GG (n=23)
SG (n=6)
LSM
SE
LSM
SE
pH45
6.20
0.02
6.31
0.04
pH24h
5.71
0.02
5.65

0.04
L* (lightness)
55.37a
0.41
52.92b 0.81
a* (Red)
14.81
0.33
16.16
0.66
b* (Yellow)
8.23
0.16
8. 55
0.31
Water loss percentage by
preservation (%)
2.35
0.12
1.94
0.23
Water loss percentage by
processing (%)
29.42
0.45
30.17
0.88
Toughness (N)
61.89
3.36

66.10
6.58
Note: means LMS in the same rows bearing different bearing superscripts were
statistically different (P<0.05).

3.2.3.2. T30N/StyI polymorphism
Difference of pH45' in (NN = 6,32, TN = 6.13, TT = 6.24)
genotypes was not statistically significant (P>0.05). However, the
difference in pH24h was statistically significant from one genotype to
anothers (P = 0.074).


24
Water loosing percentage by preservation ranged from 2.02 to
2.77%, and water loss percentage by processing was ranged from
28.74 to 29.45 %. Differences in water loosing percentage by
preservation and water loosing percentage by processing among
three genotypes were not statistically significant (P>0.05).
Table 14: Influence of T30N/StyI polymorphism on meat quality of Ban
pigs
NN (n=3)
TN (n=4)
TT (n=22)
Parameters
LSM SE
LSM SE
LSM SE
pH45’
6.32 0.06
6.13 0.05

6.24 0.02
pH24h
5.92 0.06
5.77 0.05
5.68 0.02
L* (lightness)
58,58a 1.30
56.25a 1.13
54.10b 0.48
a* (Red)
14.5 6 1.00
15.12 0.86
15.16 0.37
b* (Yellow)
7.23a 0.35
7.68a 0.31
8.56b 0.13
Water loss percentage by
preservation (%)
2.02 0.28
2.77 0.24
2.18 0.10
Water loss percentage by
processing (%)
28.74 1.26
29.41 1.09
29.45 0.46
Toughness (N)
65.52 9.52
62.64 8.25

62.41 3.52
Note: means LMS in the same rows bearing different superscripts were statistically
different (P<0.05).

No difference in (a*) red of meat among three genotypes.
However, there was a significant difference in (L*) brightness and
(b*) yellowness of meat among TT, NN and TN genotype (<0.05).
Thus T30N/StyI polymorphism influenced the brightness and
yellowness of meat.
3.2.3.3. V199I/BsaHI polymorphism
After slaughter, pH value of meat in the genotypes was rapidely
decreased, pH45' values in IV, VV genotype was 6.30 and 6.21 and
were reduced to 5.68 and 5.70, respectively after 24 hours of
slaughter. However, no effect of genotype on pH value at pH 45' and
pH24h (P>0.05). Similarly, the difference of water loss percentage
among genotypes was not statistically significant (P>0.05).
Effects of genotype at V199I/BsaHI polymorphisms point on
color of Ban pig meat was not found, (P>0.05).
The difference in toughness of meat between IV and VV
genotype was also not statistically significant (P>0.05).


25
It was concluded that the influence of gene polymorphisms on
meat quality of Lung Pu pigs varied and was not the same in different
parameters.
Table 15: Influence of V199I/BsaHI polymorphism on meat quality of
Ban pigs
IV (n=3)
VV (n=26)

Parameters
LSM
SE
LSM
SE
pH45’
6.30
0.06
6.21
0.02
pH24h
5.68
0.07
5.70
0.02
L* (lightness)
54,64
1.30
54,89
0.44
a* (Red)
14.09
0.96
15.21
0.33
b* (Yellow)
8.51
0.45
8.28
0.15

Water loosing percentage by
2.44
0.34
2.25
0.12
preservation (%)
Water loosing percentage by
29.10
1.25
29.63
0.43
processing (%)
Toughness (N)
56.37
9.27
63.50
3.15

Chapter IV
CONCLUSIONS AND RECOMMENDATIONS
4.1. Conclusion
4.1.1. Appearance characteristic, performance and meat quality of
Lung Pu and Ban pigs
- Lung Pu and Ban pigs characterised by whole black body with
6 white spots and whole black body with 4 white threads,
respectively These pigs were of majority in the population.
- Reproductive performance of Lung Pu and Ban pigs was
affected by parity and not affected by boars and year. Number and
weight of piglets/litter over parity in both breeds tended to increase
gradually from parity 1, peaked at parity of 3-4 and decreased at

parity of 5-6.
- Reproductive performance Lung Pu and Ban pigs was similar
to that of many native breeds. Number of newborn alive/litter,
number weaned piglet/litter and weaned pig weight at 45 days of age
of Lung Pu pigs were 7.42 piglets, 7.03 and 29.03 kg/litter,