Supporting Tables
Functional and safety evaluation of transgenic pork rich in omega-3 fatty acids
Maoxue Tang·Lili Qian·Shengwang Jiang·Jian Zhang·Pengkun Song·
Yaoxing Chen·Wentao Cui*·Kui Li
M. Tang·L. Qian·S. Jiang·W. Cui·K. Li
Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, 100193, P R China.
J．Zhang·P. Song
Institute for Nutrition and Food Safety, Chinese Center for Disease Control and Prevention, Beijing, 100050, P R China.
Y. Chen
College of Animal Medicine, China Agricultural University, Beijing, 100193, P R China.
*Corresponding author, E-mail:
Table S1 Absolute and relative organ weights for female mice fed with different diets
type pork diet / Low dose
sFat-1 transgenic
pork diet / High dose wild
type pork diet / High dose
sFat-1 transgenic
pork diet
Absolute weights
Body weight / 42.7±4.05a / 43.7±4.71 / 43.8±3.58 / 42.2±3.58 / 40.1±2.89
Adrenals / 0.013±0.0027 / 0.014±0.0050 / 0.014±0.0030 / 0.014±0.0023 / 0.012±0.0027
Brains / 0.34±0.051 / 0.33±0.038 / 0.34±0.075 / 0.35±0.052 / 0.33±0.037
Heart / 0.17±0.009 / 0.18±0.035 / 0.18±0.012 / 0.20±0.045 / 0.17±0.033
Liver / 1.34±0.214 / 1.40±0.107 / 1.38±0.188 / 1.36±0.154 / 1.30±0.380
Spleen / 0.14±0.022 / 0.12±0.038 / 0.13±0.027 / 0.12±0.020 / 0.13±0.041
Lung / 0.21±0.026 / 0.20±0.038 / 0.20±0.026 / 0.22±0.022 / 0.20±0.037
Kidneys / 0.38±0.043 / 0.35±0.047 / 0.36±0.025 / 0.36±0.027 / 0.34±0.022
Testes / 0.023±0.0061 / 0.023±0.0068 / 0.021±0.0057 / 0.025±0.0089 / 0.022±0.0056
Epididymis / 0.138±0.0396 / 0.126±0.0443 / 0.137±0.0636 / 0.122±0.0349 / 0.123±0.0720
pancreas / 0.127±0.0390 / 0.111±0.0224 / 0.120±0.0326 / 0.135±0.0193 / 0.122±0.0274
Thymus / 0.096±0.0183 / 0.093±0.0384 / 0.097±0.0209 / 0.088±0.0162 / 0.099±0.0175
Small intestine / 2.13±0.321 / 1.90±0.332 / 1.93±0.272 / 2.17±0.206 / 1.91±0.161
Length small int. / 57.0±5.45 / 55.8±3.20 / 55.2±3.16 / 55.7±5.84 / 55.1±4.09
Relative weights
Adrenals / 0.032±0.0076 / 0.033±0.0105 / 0.033±0.0069 / 0.034±0.0060 / 0.031±0.0080
Brains / 0.807±0.1875 / 0.765±0.1310 / 0.784±0.2095 / 0.836±0.2098 / 0.816±0.0977
Heart / 0.404±0.0496 / 0.408±0.1019 / 0.411±0.0537 / 0.482±0.1233 / 0.430±0.0948
Liver / 3.16±0.566 / 3.24±0.465 / 3.20±0.626 / 3.27±0.552 / 3.20±0.715
Spleen / 0.322±0.0540 / 0.288±0.0943 / 0.305±0.0632 / 0.279±0.0733 / 0.335±0.1131
Lung / 0.493±0.0839 / 0.463±0.0829 / 0.447±0.0492 / 0.532±0.0588 / 0.493±0.1087
Kidneys / 0.885±0.0998 / 0.807±0.1747 / 0.815±0.0306 / 0.875±0.1311 / 0.845±0.0544
Testes / 0.056±0.0162 / 0.053±0.0145 / 0.047±0.0127 / 0.060±0.0217 / 0.056±0.0134
Epididymis / 0.322±0.0825 / 0.296±0.1191 / 0.318±0.1606 / 0.288±0.0654 / 0.309±0.1867
pancreas / 0.301±0.1034 / 0.255±0.0456 / 0.278±0.0864 / 0.322±0.0585 / 0.302±0.0591
Thymus / 0.229±0.0624 / 0.217±0.0940 / 0.222±0.0453 / 0.213±0.0566 / 0.249±0.0487
Small intestine / 5.06±1.095 / 4.44±1.075 / 4.43±0.658 / 5.19±0.846 / 4.78±0.469
Length small int. / 1.35±0.170 / 1.30±0.197 / 1.27±0.144 / 1.33±0.177 / 1.38±0.123
Note: Relative organ weights expressed as g/100g body weight. Small intestinal length and relative length is expressed in cm and cm/g body weight. For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
aStatistically significantly different between the basic diet and the high dose sFat-1 transgenic pork diet
groups (p < 0.05).
Table S2a Blood biochemical findings in mice fed with different low dose diets
A / B / CFemale mice
GPT/ALT (U/L) / 24.3±4.76 / 23.8±11.36 / 22.8±6.50
GOT/AST (U/L) / 117.5±25.33 / 117.7±18.95 / 120.8±28.62
ALB (g/L) / 38.9±1.84 / 39.4±4.40 / 39.0±1.66
Cre (umol/L) / 64.8±14.95 / 68.0±13.82 / 79.4±33.38
BUN (mmol/L) / 6.2±1.04 / 6.0±0.91 / 6.2±0.87
TP (g/L) / 65.1±3.14 / 68.0±3.58 / 67.4±4.13
TC (mmol/L) / 2.5±0.58 / 2.8±0.27 / 2.6±0.49
TG (mmol/L) / 1.0±0.48 / 1.0±0.34 / 1.0±0.41
HDL_C (mmol/L) / 1.9±0.47 / 2.2±0.44 / 1.9±0.31
Male mice
GPT/ALT (U/L) / 26.5±16.96 / 23.5±11.64 / 23.5±5.79
GOT/AST (U/L) / 109.2±14.17 / 115.4±64.02 / 114.3±37.76
ALB (g/L) / 35.3±1.59 / 35.8±2.69 / 36.1±1.76
Cre (umol/L) / 50.2±5.61a / 49.9±10.94 / 56.4±5.91
BUN (mmol/L) / 7.7±1.50a / 8.2±1.78 / 9.0±1.51
TP (g/L) / 64.4±6.73 / 64.2±6.13 / 65.5±4.17
TC (mmol/L) / 3.7±1.08 / 3.9±0.66 / 4.0±0.70
TG (mmol/L) / 1.2±0.43 / 1.2±0.62 / 1.1±0.29
HDL_C (mmol/L) / 2.8±0.79 / 2.8±0.60 / 3.0±0.46
Note: A: Basic diet; B: Low dose wild type pork diet; C: Low dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
aStatistically significantly different between the basic diet and the sFat-1 transgenic pork diet groups (p < 0.05).
Table S2b Blood biochemical findings in mice fed with different high dose diets
Female mice
GPT/ALT (U/L) / 24.3±4.76 / 20.3±11.44 / 22.2±13.24
GOT/AST (U/L) / 117.5±25.33 / 104.9±20.90 / 99.8±25.40
ALB (g/L) / 38.9±1.84 / 39.4±2.78 / 38.4±2.99
Cre (umol/L) / 64.8±14.95 / 74.2±31.05 / 68.8±32.05
BUN (mmol/L) / 6.2±1.04aa / 8.5±2.47 / 8.2±0.90
TP (g/L) / 65.1±3.14 / 68.3±4.56 / 65.6±6.64
TC (mmol/L) / 2.5±0.58 / 2.9±0.58 / 2.8±0.80
TG (mmol/L) / 1.0±0.48 / 0.9±0.24 / 0.9±0.16
HDL_C (mmol/L) / 1.9±0.47 / 2.2±0.45 / 2.1±0.56
Male mice
GPT/ALT (U/L) / 26.5±16.96 / 24.6±12.38 / 25.3±18.18
GOT/AST (U/L) / 109.2±14.17 / 113.1±21.81 / 108.3±31.23
ALB (g/L) / 35.3±1.59 / 35.6±2.78 / 35.8±1.46
Cre (umol/L) / 50.2±5.61 / 53.1±12.81 / 50.4±7.99
BUN (mmol/L) / 7.7±1.50a / 9.3±1.82 / 9.5±1.92
TP (g/L) / 64.4±6.73 / 64.9±3.06 / 64.0±3.05
TC (mmol/L) / 3.7±1.08 / 4.3±0.76 / 3.9±0.45
TG (mmol/L) / 1.2±0.43 / 1.1±0.44 / 1.1±0.33
HDL_C (mmol/L) / 2.8±0.79 / 3.1±0.52 / 2.9±0.31
Note: A: Basic diet; B: High dose wild type pork diet; C: High dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
aStatistically significantly different between the basic diet and the sFat-1 transgenic pork diet groups (p < 0.05), aa p < 0.01.
Table S3a Haematological findings in female mice fed with different low dose diets
A / B / CWBC (109/L) / 3.6±1.03 / 3.8±1.46 / 3.9±0.89
RBC (1012/L) / 10.5±2.32 / 11.0±1.03 / 10.5±0.77
HB (g/L) / 145.3±42.10 / 151.9±19.67 / 152.0±10.45
MCV (fL) / 46.9±2.51 / 46.3±1.16 / 47.0±1.70
RDW (%) / 14.3±1.10 / 13.8±0.77 / 13.8±0.80
PLT (109/L) / 585.8±115.13 / 578.1±146.30 / 571.4±110.82
MPV (fL) / 8.0±0.93 / 7.4±0.78 / 7.3±0.89
PCT (%) / 0.4±0.09 / 0.5±0.05b / 0.4±0.06
PDW (%) / 0.5±0.83 / 0.7±0.67 / 0.9±0.98
GRAN (109/L) / 3.0±0.77 / 3.1±1.24 / 3.2±1.37
LYM (109/L) / 0.4±0.14 / 0.4±0.17 / 0.5±0.39
MON (109/L) / 0.5±0.08 / 0.4±0.15 / 0.5±0.21
GRA% (%) / 75.8±9.79 / 80.8±5.73 / 76.5±6.48
LYM% (%) / 10.2±3.21 / 10.9±3.27 / 11.8±3.95
MON% (%) / 14.0±6.96 / 9.1±2.46 / 11.7±4.87
Note: A: Basic diet; B: Low dose wild type pork diet; C: Low dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
bStatistically significantly different between the wild type pork diet and sFat-1 transgenic pork diet groups (p < 0.05).
Table S3b Haematological findings in female mice fed with different high dose diets
A / B / CWBC (109/L) / 3.6±1.03 / 3.2±1.80 / 3.4±1.43
RBC (1012/L) / 10.5±2.32 / 9.4±2.58b / 11.3±1.89
HB (g/L) / 145.3±42.10 / 158.4±45.54 / 158.5±34.83
MCV (fL) / 46.9±2.51 / 45.1±1.79 / 46.0±2.67
RDW (%) / 14.3±1.10 / 14.0±1.05 / 13.8±0.71
PLT (109/L) / 585.8±115.13 / 536.8±115.05 / 559.8±129.32
MPV (fL) / 8.0±0.93 / 7.3±1.24 / 7.6±0.95
PCT (%) / 0.4±0.09 / 0.4±0.08 / 0.4±0.13
PDW (%) / 0.5±0.83 / 0.7±0.81 / 0.4±0.37
GRAN (109/L) / 3.0±0.77 / 2.6±1.08 / 2.5±1.05
LYM (109/L) / 0.4±0.14 / 0.4±0.22 / 0.4±0.23
MON (109/L) / 0.5±0.08 / 0.4±0.19 / 0.4±0.17
GRA% (%) / 75.8±9.79 / 76.9±3.38bb / 81.0±4.08
LYM% (%) / 10.2±3.21 / 10.6±2.32b / 8.4±2.14
MON% (%) / 14.0±6.96 / 12.4±2.34b / 10.8±2.52
Note: A: Basic diet; B: High dose wild type pork diet; C: High dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
bStatistically significantly different between the wild type pork diet and the sFat-1 transgenic pork diet groups (p < 0.05), bb p < 0.01.
Table S3c Haematological findings in male mice fed with different low dose diets
A / B / CWBC (109/L) / 4.0±1.98 / 4.0±1.66 / 4.0±1.37
RBC (1012/L) / 10.4±3.59 / 10.9±1.08 / 10.2±0.88
HB (g/L) / 134.7±37.98 / 139.2±31.95 / 140.7±8.92
MCV (fL) / 45.1±1.60 / 44.3±1.25 / 45.3±1.57
RDW (%) / 14.0±0.51 / 14.2±0.75 / 14.1±0.71
PLT (109/L) / 671.5±204.16 / 663.2±174.38 / 661.8±144.38
MPV (fL) / 7.6±1.21 / 8.0±1.74 / 7.6±1.29
PCT (%) / 0.5±0.21 / 0.5±0.12 / 0.5±0.08
PDW (%) / 0.7±0.65 / 0.8±0.49 / 1.0±0.86
GRAN (109/L) / 2.4±0.92 / 2.8±1.28 / 2.4±1.05
LYM (109/L) / 0.8±0.75 / 0.7±0.17 / 0.9±0.44
MON (109/L) / 0.6±0.43 / 0.5±0.18 / 0.6±0.28
GRA% (%) / 65.0±11.46 / 70.4±8.98 / 66.0±9.12
LYM% (%) / 17.1±7.37 / 15.2±4.75 / 18.0±6.42
MON% (%) / 17.9±6.08 / 14.4±5.30 / 15.0±2.87
Note: A: Basic diet; B: Low dose wild type pork diet; C: Low dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
Table S3d Haematological findings in male mice fed with different high dose diets
A / B / CWBC (109/L) / 4.0±1.98 / 3.6±1.88 / 3.8±1.48
RBC (1012/L) / 10.4±3.59 / 8.9±3.65 / 9.0±2.20
HB (g/L) / 134.7±37.98 / 145.2±64.10 / 143.5±36.90
MCV (fL) / 45.1±1.60 / 44.4±2.84 / 44.6±1.51
RDW (%) / 14.0±0.51a / 14.3±1.10 / 14.8±0.89
PLT (109/L) / 671.5±204.16 / 531.3±146.86 / 632.4±185.89
MPV (fL) / 7.6±1.21 / 7.4±0.94 / 7.2±1.27
PCT (%) / 0.5±0.21 / 0.4±0.13 / 0.5±0.21
PDW (%) / 0.7±0.65 / 1.4±1.29 / 1.2±0.79
GRAN (109/L) / 2.4±0.92 / 2.1±1.25 / 2.7±1.57
LYM (109/L) / 0.8±0.75 / 0.7±0.50 / 0.6±0.31
MON (109/L) / 0.6±0.43 / 0.6±0.43 / 0.4±0.21
GRA% (%) / 65.0±11.46a / 64.4±10.15bb / 74.5±7.31
LYM% (%) / 17.1±7.37 / 17.8±5.23b / 13.0±5.37
MON% (%) / 17.9±6.08 / 17.7±5.88b / 14.8±3.55
Note: A: Basic diet; B: High dose wild type pork diet; C: High dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
aStatistically significantly different between the basic diet and the sFat-1 transgenic pork diet groups (p < 0.05).
bStatistically significantly different between the sFat-1 transgenic pork diet and the wild type pork groups (p < 0.05), bb p < 0.01.
Table S4 Serum immunoglobulin concentrations in mice fed with different low dose diets
Group / IgE (mg/L) / IgM (mg/L) / IgA (mg/L) / IgG (mg/L)Female mice
A / 1.4±0.17 / 11.5±3.86 / 1.3±0.86 / 6.4±4.16
B / 1.5±0.14 / 12.2±5.14 / 1.0±0.38 / 6.4±2.63
C / 1.4±0.20 / 10.7±4.12 / 1.2±0.59 / 6.0±1.75
Male mice
A / 1.4±0.20 / 10.2±2.36 / 1.4±0.77 / 5.9±2.97
B / 1.3±0.13 / 11.9±3.98 / 0.9±0.35 / 5.3±1.43
C / 1.3±0.18 / 11.0±4.02 / 1.2±0.88 / 5.1±1.15
Note: A: Basic diet; B: Low dose wild type pork diet; C: Low dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
Table S5 Distribution of T lymphocyte subsets in peripheral blood from mice fed with different low dose diets
Female mice
A / 43.6±11.42 / 22.8±4.92 / 2.0±0.85
B / 44.8±6.89 / 20.4±5.06 / 2.3±0.73
C / 49.2±8.44 / 19.4±4.49 / 2.7±0.92
Male mice
A / 39.3±6.67a / 16.0±4.21 / 2.6±0.68
B / 32.8±6.24bb / 15.0±2.10 / 2.2±0.59
C / 43.6±4.58 / 18.0±5.82 / 2.7±1.03
Note: A: Basic diet; B: Low dose wild type pork diet; C: Low dose sFat-1 transgenic pork diet.
For statistical analysis, sFat-1 transgenic pork diet was compared to wild type pork diet and basic diet, respectively. Data are presented as mean ± SD.
aStatistically significantly different between the basic diet and the sFat-1 transgenic pork diet groups (p < 0.05).
bbStatistically significantly different between the sFat-1 transgenic pork diet and the wild type pork groups (p < 0.01).
Table S6 Bacterial counts in day 15 and 30 faecal samples from female mice fed with different low dose diets
Group / Total aerobe / Total anaerobe / Lactobacilli / Bifidobacteria / Streptococcusthermophilus / Bacteroides / Coliforms / Enterococci / Clostridia / Staphylococcus
aureus
Bacterial counts-Day 15
A / 6.8±0.22 / 7.6±0.98 / 8.2±0.33 / 6.2±1.73 / 7.8±0.31 / 2.6±0.56 / 4.7±0.74 / 4.2±1.39 / <1 / 2.6±0.52
B / 6.8±0.56 / 8.4±0.61 / 8.2±0.43 / 7.3±1.54 / 7.6±0.47 / 3.0±0.37 / 4.5±0.94 / 4.2±0.76 / <1 / 2.6±0.58
C / 7.0±0.46 / 8.2±0.28 / 7.9±0.54 / 7.4±0.67 / 7.4±0.61 / 2.6±0.50 / 4.6±0.80 / 4.9±0.70 / <1 / 3.0±0.59
Bacterial counts-Day 30
A / 7.4±0.74 / 8.2±0.54 / 9.2±0.20 / 8.9±0.28 / 6.8±0.44 / 2.7±0.54 / 4.8±0.64 / 6.0±1.08 / <1 / 2.8±0.49
B / 7.8±0.53 / 7.7±0.95 / 8.4±0.32 / 8.1±0.55 / 6.8±0.69 / 2.8±0.56 / 5.3±1.55 / 5.9±1.17 / <1 / 2.8±0.50
C / 7.7±0.63 / 8.5±0.27 / 8.9±0.44 / 8.7±0.41 / 7.8±0.97 / 2.9±0.52 / 5.1±0.65 / 6.5±0.70 / <1 / 2.6±0.56
Note: A: Basic diet; B: Low dose wild type pork diet; C: Low dose sFat-1 transgenic pork diet.