Children with protein-energy malnutrition (PEM) need a food product that can be easily absorbed by the body because they have impaired nutrient utilization and lipid profile disorder. The provision of fermented products such as tempeh is an alternative approach. Seeds of tree bean (
Protein-energy malnutrition (PEM) is defined as malnutrition in which energy and protein intake are under the nutritional adequacy rate (
The highest prevalence of PEM in children under 5 years of age (
Undernutrition and the presence of infectious diseases can aggravate the diseases and worsen nutritional status. Poor nutrition in children leads to slow growth and affects cognitive abilities (
Children with PEM need food products that are easily absorbed by the body because of impaired utilization of nutrients. Fermented foods contain nutrients that are easily absorbed by the intestines; one of the fermented foods is tempeh (
Tree bean (
Tempeh flour from tree bean (
This study is included in the research project Study of the Administration of Tempeh Flour from Tree Bean (
This study used tempeh flour from tree bean seeds as samples. Tree bean seeds (Figure
Tree bean (
Male Wistar rats (8 – 12 weeks old, 150 – 200 g, n = 24), were acquired from the Centre for Food and Nutrition of Universitas Gadjah Mada, Yogyakarta, Indonesia.
Albumin and hemoglobin level measurements as criteria of PEM were performed after 14 days of zero-protein diet administration. Measurements of cholesterol, triglyceride, LDL, and HDL levels were performed before the intervention and at the end of the intervention. Blood sampling was done to analyze albumin, hemoglobin, cholesterol, triglyceride, LDL, and HDL through the plexus retroorbital. Examination of triglyceride used Glycerol 3 Phospate Oxidase-Peroxidase Amino Phenazone (GPOAP) method, meanwhile cholesterol, LDL and HDL used Cholesterol Oxidase-Peroxidase Aminoantipyrine Phenol (CHOD-PAP) with enzymatic spectrophotometric basis. The atherogenic index of plasma (AIP) was calculated from the log (triglyceride/HDL). Blood serum was analyzed in the Centre for Food and Nutrition of Universitas Gadjah Mada, Yogyakarta, Indonesia.
The seeds were washed and soaked in water for 2 h. Then, the soaking water was removed, and the seeds were washed thoroughly. Tree bean seeds were boiled in water for 2 h. Then, the water was removed and tree bean seeds were drained. Tree bean seeds were peeled (the black part) as seen in Figure
Peeling the black part of tree bean seeds.
Inoculation.
Tempeh from tree bean seeds after fermentation 48 h.
Tempeh from tree bean seeds was sliced using a slicer and then blanched with hot steam for 2 min. Next, tempeh was dried by oven at 60 °C for 6 h (Figure
Drying tempeh.
Tempeh flour from tree bean seeds.
The animals were individually housed and provided a standard feed of Comfeed II at 15 g per rat per day and water ad libitum. Rats were randomly divided into four groups (n = 6/group): the normal control group (K+), the PEM group without treatment (K−), the group fed tempeh flour from tree bean (
Statistical analyses were performed using the IBM SPSS Statistic 22 software. Data are presented as mean ±SD or median. A paired
The average body weight of the animals significantly increased in all the groups (
The average body weight before and after treatments.
Groups | Bodyweight (g) | |||
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Pre | Post | Δ |
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K+ | 124(120-128) | 156(154-161) | 33(31-35) | 0.026* |
K- | 99(97-104) | 109(107-113) | 10(7-12) | 0.027* |
P1 | 100(99-103) | 130(128-132) | 29(29-31) | 0.020* |
P2 | 102.5(98-105) | 129(126-132) | 27.5(25-28) | 0.026* |
|
0.002* | 0.000* | 0.000* |
Note: normal control group (K+), PEM group without treatment (K-), tempeh flour from seeds of
The results of collaboration research showed that the rats acquired PEM after the administration of the zero-protein diet for 14 days. Rats achieved PEM with albumin (1.21 – 1.33 g.dL-1) and hemoglobin (11.86 – 12.13 g.dL-1) (
A significant difference in triglyceride was found after the administration of tempeh flour from tree bean seeds in the K+ (
Triglycerides level before and after the administration of tempeh flour from tree bean seeds.
Groups | Triglycerides (mg.dL-1) | |||
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Pre | Post | Δ |
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67.8 ±2.089b,c,d | 69.29 ±2.58b,c,d | 1.44 ±0.517c | 0.001* | |
105 ±3,7276a | 109 ±3.88a,c,d | 3.87 ±3.32c | 0.036* | |
105 ±2,888a | 80.82 ±2.46a,b,d | -24.45 ±3.68a,b,d | 0.001* | |
103 ±5,83a | 95.61 ±3.059a,b,c | -7.94 ±784c | 0.056* | |
0.001* | 0.001* | 0.001* |
Note: normal control group (K+), PEM group without treatment (K-), tempeh flour from seeds of
Total cholesterol levels before and after the administration of tempeh flour from tree bean seeds.
Groups | Total cholesterol levels (mg.dL-1) | |||
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Pre | Post | Δ |
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74.82(71.53-76.64) | 76.57(72.8-78.66)b,c,d | 1.75(0.97-2.54)c,d | 0.028* | |
1.56(151.82-167.88) | 1.59(155.65-168.2)a,c,d | 2.2(0.32-8.01)c,d | 0.028* | |
1.50(146.72-155.47) | 93.31(91.21-97.91)a,b,d | -55.44(-64.26-(-51.73))a,b | 0.028* | |
1.54(146.28-157.66) | 1.03(99.58-107.11)a,b,c | -49.04(-57.24-(-42.53))a,b | 0.028* | |
0.001* | 0.001* | 0.001* |
Note: normal control group (K+), PEM group without treatment (K-), tempeh flour from seeds of
The highest decrease in total cholesterol occurred in the P1 group, followed by that in the P2 group. LDL levels in the K+ (
LDL levels before and after treatments tempeh flour from tree bean seeds.
Groups | LDL levels (mg.dL-1) | |||
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Pre | Post | Δ | P | |
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21.11(18.69-23.53) | 22.38(20.98-25.17)b,c,d | 0.94(0.22-3.69)c,d | 0.028* | |
56.05(54.67-58.13) | 57.34(53.85-59.44)a,c,d | 0.59(-0.82 -1.99)c,d | 0.172 | |
56.06(53.29-57.44) | 27.62(23.78-30.77)a,b,d | -27.74(-33.66-(-25.98))a,b,d | 0.028* | |
55.02(53.98-60.9) | 36.01(33.57-37.76)a,b,c | -20.05(-25.24-(-16.22))a,b,c | 0.028* | |
0.004* | 0.001* | 0.001* |
Note: normal control group (K+), PEM group without treatment (K-), tempeh flour from seeds of
HDL levels before and after treatments tempeh flour from tree bean seeds.
Groups | HDL levels (mg.dL-1) | |||
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Pre | Post | Δ |
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79.25(76.87-81.63) | 77.775(72.03-80.46)b.c.d | -1.34(-6.2-(-0.24))c.d | 0.028* | |
46.26(44.22-51.02) | 46.36(43.68-48.28)a.c.d | -0.5(-2.74-1.16)c.d | 0.249 | |
45.24(42.86-47.62) | 66.285(64.37-70.5)a.b.d | 21.55(20.32-22.88)a.b.c | 0.028* | |
44.9(42.86-47.62) | 58.62(55.17-61.3)a.b.c | 12.57(9.85-18.44)a.b.d | 0.028* | |
0.003* | 0.001* | 0.001* |
Note: normal control group (K+), PEM group without treatment (K-), tempeh flour from seeds of
Atherogenic index of plasma (AIP) in K+ (
AIP before and after treatments tempeh flour from tree bean seeds.
Groups | AIP (mg.dL-1) | |||
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Pre | Post | Δ |
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-0.07±0.01 | -0.048±0.03 | 0.02(0.01-0.05) | 0.048* | |
0.35±0.01 | 0.38±0.01 | 0.03±0.01 | 0.001* | |
0.36(0.35-0.41) | 0.09±0.02 | -0.28±0.01 | 0.027* | |
0.36(0.34-0.42) | 0.21±0.03 | -0.13(-0.25-(-0.11)) | 0.027* | |
0.004* | 0.000* | 0.000* |
Note: normal control group (K+), PEM group without treatment (K-), tempeh flour from seeds of
Total cholesterol, triglyceride, and LDL levels increased in rats with PEM; meanwhile, HDL levels decreased compared to the normal control group. Chronic malnutrition causes endocrine changes leading to metabolic profile disorders. Previous studies demonstrated that 98.9% of children have dyslipidemia with low HDL and hypertriglyceridemia (
Tree bean seeds in this study were processed into fermented food as tempeh (
In this study, the administration of tempeh flour from tree bean seeds reduced triglycerides, cholesterol, and LDL levels and increased HDL levels in PEM rats. These results are consistent with previous studies, in which, the administration of tempeh flour decreased total cholesterol in high-saturated fat-diet induced rats (
The content of essential amino acids found in tree bean seeds also plays a role in reducing triglyceride, cholesterol, and LDL levels in PEM rats. A previous study explained that the provision of essential amino acid supplements and phytosterols are taken 3 times a day for 4 weeks can reduce triglyceride, cholesterol, and LDL levels in the subject (
Tree bean seeds contain a thioproline compound that produces a distinctive odor in tree bean seeds. Thioproline contains amino acids and is a natural metabolite that acts as an intracellular sulfhydryl antioxidant and a free radical scavenger and protects cell membranes from damage caused by oxygen derivatives (
The administration of tempeh flour from tree bean (
We would like to thank the Faculty of Medicine, Universitas Diponegoro, for funding this research.
This study was supported by funding received from the Faculty of Medicine, Universitas Diponegoro 2020.
The authors declare no conflict of interest.
The experiments were approved by The Ethical Committee of Medical Research of the Faculty of Medicine, Universitas Diponegoro (no. 22/EC/H/FKUNDIP/IV/2020), Indonesia.