To determine whether dried fermented ginger (DFG), fermented with Japanese mugwort silage juice, could be replaced by fermented corncob powder (FCP) as a of feed ingredient source without significant body weight decrease or damage to visceral organs (using gross anatomical observation), to intestinal villi (using light microscopy), or to the epithelial cells on the villus apical surface (using scanning electron microscopy) the following investigation was performed. Sixty-four male broilers were allotted to 4 groups: a basal diet group (control group), and basal diet groups with DFG at a level of 50 ppm; with DFG at 50 ppm and FCP at 250 ppm (50 ppm DFG + 250 ppm FCP group); and with FCP at a level of 500 ppm (500 ppm FCP group). Feed intake, body weight gain, feed efficiency, carcass quality, small intestinal length and weight, and visceral organ weight were not different among groups. Furthermore, regarding intestinal villus height, villus area and crypt depth, a significant difference was not found among the groups. When these values of the control were expressed as an index of 100, the duodenal villus height of the 50 ppm DFG + 250 ppm FCP group and the 500 ppm FCP group were 114 and 119, respectively. The duodenal villus area of the 50 ppm DFG + 250 ppm FCP group and the 500 ppm FCP group were 125 and 158, respectively. These villus heights and areas are thought to be activated. On the epithelial cells on the villus apical surface in the duodenum and jejunum, the 50 ppm DFG + 250 ppm FCP group had protuberated cells into the intestinal lumen and deeper cells at the sites of recently exfoliated cells, suggesting that these cells are activated. The present results indicate that small amounts of fermented corncob powder can be used as a feed supplement when mixed with fermented ginger powder, due to the synergy between the two ingredients, resulting in a 6% increase in body weight gain。
Feed represents the largest cost associated with poultry production, and the major source of feed ingredients is corn. However, corn ethanol production has grown rapidly in the last decade, subsequent to corn’s first being employed in biogas production [
Japanese mugwort silage juice (JMS) liquid includes natural microorganisms, such as lactic acid bacteria, yeast fungus, photosynthetic bacteria, ray-fungus, hyperthermal bacteria, and Aspergillus and Bacillus subtilis [
The aim of the present study was to determine whether ginger could be replaced by corncob as a source of feed ingredient to economize feed costs without a significant decrease in body weight. The following observations were performed: the presence or absence of damage on visceral organs using gross anatomical observation; on intestinal villi using light microscopy; and on the epithelial cells on the villus apical surface using scanning electron microscopy.
Japanese mugwort plants were harvested and ensiled at room temperature to get JMS. Grounded ginger by-product was added into the JMS solution, kept under anaerobic conditions at room temperature for 4 - 5 d, dried in a hot air oven at 50˚C for 1 - 2 d and again ground for finally making DFG [
Newly hatched male broilers (Marshall Chunky) were obtained from a commercial hatchery, and conventional starter (1 - 21 d) and finisher (22 - 49 d) diets were used as basal diets (
At 49 d of age, 4 birds per group from each replicate were dissipated. The entire small intestine was then removed and processed in a mixture of 3% glutaraldehyde and 4% paraformaldehyde fixative solution in 0.1 M cacodylate buffer (pH 7.4). The same fixative solution was additionally infused into the intestinal lumen at low pressure. The duodenum extended from the gizzard to the pancreatic and bile ducts; the jejunum extended from the bile ducts to Meckel’s diverticulum, and the ileum from the diverticulum to the ileocecal junction. A 2-cm length of the middle part of each intestinal segment was excised and prepared for light and scanning electron microscopy (SM). Each segment was gently flushed with 0.1 M phosphate-buffered saline (pH 7.4) to remove the intestinal contents.
Samples for SM were cut lengthwise along the line of the mesentery, opened, and flushed with 0.1 M phosphate buffered saline (pH 7.4). These samples were
Item | Starter 1 to 21 d | Finisher 22 to 49 d |
---|---|---|
Ingredient | ||
Corn | 590 | 420 |
Milo | 20 | 220.0 |
Corn gluten meal | 290 | - |
Soybean meal (45%CP) | - | 270.0 |
Rice bran | 23.0 | 52.0 |
Fish meal (57%CP) | 70.0 | 30.0 |
Tallow | 5.0 | 6.0 |
Vitamin/mineral premix1 | 2.0 | 2.0 |
Total (g) | 1000.00 | 1000.00 |
Calculated composition | ||
Crude protein (%) | 22.00 | 18.00 |
ME (kcal/kg) | 3,050 | 3,250 |
Crude fat (%) | 4 | 6 |
Crude fiber (%) | 4 | 4 |
Crude ash (%) | 7 | 7 |
Calcium (%) | 0.8 | 0.7 |
Available Phosphorus (%) | 0.5 | 0.45 |
1Vitamin and mineral premix include per kg of diet: retinyl acetate, 2880 µg; cholecalciferol, 48 µg; DL-α- tocopherol acetate, 35 mg; menadione, 2.6 mg; thiamine, 5.8 mg; riboflavin, 7.3 mg; pyridoxine, 10.4 mg; cobalamine, 12.6 µg; biotin, 0.2 mg; folic acid, 1.0 mg; pantothenic acid, 16.1 mg; niacin, 69.1 mg; choline, 1400 mg; manganese, 92.4 mg; zinc, 79.9 mg; copper, 12.8 mg.
Items | DFG | FCP |
---|---|---|
Dry matter | 87.44 | |
Crude protein | 7.29 | |
Crude fat | 6.51 | |
Crude fiber | 22.10 | |
Crude ash | 13.40 | |
Calcium | 0.46 | |
Phosphorus | 0.84 | |
Gross energy (kcal/kg) | 3908 |
All parameters of DFG samples were determined in triplicate (Incharoen and Yamauchi, 2009; Incharoenet et al. 2010).
pinned flat to prevent curling and fix them vertically with the mucosal face downward in a mixture of 3% glutaraldehyde and 4% paraformaldehyde in 0.1 M cacodylate buffer (pH 7.4) at room temperature for 1 h. The tissue block was cut into 5 × 6 mm square and fixed for an additional 1 h. The pieces were rinsed with 0.1 M sodium cacodylate buffer (pH 7.4) and post-fixed with 1% osmiumtetroxide in the ice-cold buffer for 2 h. Then, these pieces were washed in deionizing distilled water, stored in 45% ethanol for 24 h, and kept in 70% ethanol. Just before drying, specimens were transferred to 80%, 90% and 100% ethanol solution (120, 60 and 15 min, respectively) and submerged in t-butyl alcohol (15 min; 3 times). Afterwards, these specimens were freeze-dried (Hitachi ES-2030 freeze dryer, Hitachi Ltd., Tokyo, Japan). The freeze-dried specimens were mounted on aluminum stubs with electrically conducting carbon paste, coated (E-1030 ion sputters, Hitachi Ltd., Tokyo, Japan), and viewed under a scanning electron microscope (Hitachi S-4300SE/N, Hitachi Ltd., Tokyo, Japan) at eight kV. Morphological alterations of the epithelial cells on the villus apical surface were compared.
After keep in Bouin’s solution, the intestinal segments were embedded in paraplast wax, cut into 4-µm transverse sections, and stained with hematoxylin-eosin. Eight sections per intestinal segment per bird were used to measure villus height, villus area and crypt depth by an image analyzer (Nis-Element D, Nikon Co., Tokyo, Japan). The 48 villi per bird were randomly chosen to measure villus height, from the villus tip to the base. And the 4 mean villus heights from 4 birds were expressed as a mean villus height for 1 group. The villus area was calculated from the villus height, basal width, and apical width [
At 50 d of age, 8 birds in each group were used to measure carcass quality, small intestinal length and weight, and visceral organ weight. After decapitation, the entirety of the visceral organs was removed. The weights of the visceral organs, proventriculus, gizzard, and liver were measured. The whole small intestine, from the gizzard to the large intestine, was removed. The weights and lengths of duodenum, jejunum, and ileum were recorded.
Growth performance such as body weight and feed intake were measured weekly to calculate body weight gain, and feed efficiency was calculated from body weight gain per feed intake. Growth performance, carcass percentage, and light microscopic parameters were analyzed by a one-way analysis of variance (ANOVA). Significance was determined at p < 0.05, using Duncan’s multiple range tests to compare differences among groups.
The experiment was performed in accordance with the guidelines and rules of care and use of laboratory animal experimentation established by Kagawa University in Japan. The experimental procedure was approved by the Animal Research Committee of the Kagawa University, Japan.
Feed intake, body weight gain, and feed efficiency did not differ among the groups (
Compared with the control, the percentage weights of carcass, thigh (drumsticks and thighs), breast, wing and abdominal fat did not differ among the experimental groups (
Compared with the control, the intestinal villus height, villus area, and crypt depth in each intestinal segment of the experimental groups did not differ (
Although the majority of epithelial cells on the duodenal villus apical surface of the control group were flat (small arrows in
Items | Control | 50ppm DFG | 50 ppm DFG + 250 ppm FCP | 500ppm FCP | p-value |
---|---|---|---|---|---|
Body weight gain (g) Ratio for 100 index of control | 3102.39 ± 466.41 100 | 3217.13 ± 156.68 104 | 3301.94 ± 209.45 106 | 3115.37 ± 172. 100 | 0.725 100 |
Feed intake (g) Ratio for 100 index of control Feed efficiency Ratio for 100 index of control | 5952.08 ± 465.31 100 0.52 ± 0.05 100 | 5964.82 ± 401.23 100 0.54 ± 0.01 104 | 6090.71 ± 133.38 102 0.54 ± 0.04 104 | 5822.68 ± 621.86 98 0.54 ± 0.04 104 | 0.863 0.851 |
DFG; dried fermented ginger, FCP; fermented corncob powder.
Items | Control | 50 ppm DFG | 50 ppm DFG + 250 ppm FCP | 500 ppm FCP | p-value |
---|---|---|---|---|---|
Carcass (%) | 81.653 ± 1.530 100 | 81.023 ± 1.551 9999 | 82.676 ± 1.955 101 | 82.657 ± 0.984 101 | 0.111 |
Thigh (drumsticks and thighs) (%) | 7.802 ± 0.386 100 | 7.549 ± 0.472 97 | 7.951 ± 0.650 102 | 7.502 ± 0.700 96 | 0.355 |
Breast (%) | 19.590 ± 1.121 100 | 20.995 ± 1.702 107 | 20.407 ± 1.113 104 | 21.124 ± 1.557 108 | 0.135 |
Wing (%) | 21.775 ± 1.426 100 | 21.493 ± 1.878 99 | 22.427 ± 1.166 103 | 22.482 ± 2.115 103 | 0.578 |
Abdominal fat (%) | 1.657 ± 0.459 100 | 1.445 ± 0.396 87 | 1.481 ± 0.305 89 | 1.452 ± 0.527 88 | 0.730 |
Length, cm/100 g BW | |||||
Duodenum | 0.973 ± 0.224 100 | 1.019 ± 0.171 105 | 0.974 ± 0.768 100 | 1.024 ± 0.200 105 | 0.897 |
Jejunum | 2.599 ± 0.516 100 | 2.584 ± 0.504 99 | 2.393 ± 0.188 92 | 2.598 ± 0.375 100 | 0.710 |
Ileum | 2.571 ± 0.541 100 | 2.665 ± 0.468 104 | 2.550 ± 0.313 99 | 2.623 ± 0.383 102 | 0.951 |
Total small intestinal length | 6.143 ± 1.234 100 | 6.268 ± 1.095 102 | 5.917 ± 0.494 96 | 6.245 ± 0.792 102 | 0.876 |
Weight (g/100 g BW) | |||||
Total visceral organ | 12.944 ± 1.362 100 | 13.033 ± 1.280 101 | 11.606 ± 1.747 90 | 11.973 ± 0.618 92 | 0.097 |
Gizzard | 1.239 ± 0.110 100 | 1.295 ± 0.245 105 | 1.209 ± 0.124 98 | 1.222 ± 1.822 99 | 0.767 |
Proventriculus | 0.307 ± 0.049 100 | 0.303 ± 0.053 99 | 0.304 ± 0.037 99 | 0.245 ± 0.110 80 | 0.220 |
Liver | 2.065 ± 0.332 100 | 1.999 ± 0.294 97 | 1.927 ± 0.253 93 | 1.901 ± 0.273 92 | 0.671 |
Duodenum | 0.566 ± 0.092 100 | 0.551 ± 0.082 97 | 0.599 ± 0.103 106 | 0.581 ± 0.126 103 | 0.807 |
Jejunum | 1.291 ± 0.197 100 | 1.228 ± 0.231 95 | 1.234 ± 0.125 96 | 1.295 ± 0.194 100 | 0.836 |
Ileum | 0.945 ± 0.167 100 | 1.019 ± 0.105 108 | 0.971 ± 0.109 103 | 0.946 ± 0.137 100 | 0.645 |
DFG; dried fermented ginger, FCP; fermented corncob powder.
flat cells (small arrow) and deeper cells at the sites of recently exfoliated cells (arrows with D). The 50 ppm DFG + 250 ppm FCP (
Item | Control | 50 ppm DFG | 50 ppm DFG + 250 ppm FCP | 500 ppm FCP | P-value |
---|---|---|---|---|---|
Duodenum | |||||
Villus height | 1.32 ± 0.16 100 | 1.40 ± 0.11 106 | 1.51 ± 0.12 114 | 1.57 ± 0.10 119 | 0.534 |
Villus area | 0.12 ± 0.02 100 | 0.18 ± 0.01 150 | 0.15 ± 0.01 125 | 0.19 ± 0.03 158 | 0.227 |
Crypt depth | 385.83 ± 4.16 100 | 330.08 ± 60.48 86 | 224.76 ± 21.62 58 | 270.60 ± 24.37 70 | 0.115 |
Jejunum | |||||
Villus height | 1.21 ± 0.15 100 | 1.15 ± 0.12 95 | 1.15 ± 0.09 95 | 1.40 ± 0.03 116 | 0.226 |
Villus area | 0.14 ± 0.04 100 | 0.15 ± 0.03 107 | 0.14 ± 0.01 100 | 0.15 ± 0.03 107 | 0.987 |
Crypt depth | 216.63 ± 29.93 100 | 214.49 ± 45.51 99 | 186.76 ± 14.97 86 | 219.86 ± 14.06 101 | 0.406 |
Ileum | |||||
Villus height | 0.67 ± 0.08 100 | 0.61 ± 0.10 91 | 0.63 ± 0.10 94 | 0.67 ± 0.13 100 | 0.757 |
Villus area | 0.06 ± 0.01 100 | 0.05 ± 0.01 83 | 0.05 ± 0.01 83 | 0.07 ± 0.01 117 | 0.103 |
Crypt depth | 150.38 ± 16.49 100 | 135.33 ± 14.87 90 | 128.44 ± 13.38 85 | 136.23 ± 20.56 91 | 0.339 |
DFG; dried fermented ginger, FCP; fermented corncob powder.
As epithelial cells originate in the crypts by mitosis and migrate steadily toward the villus tips where cells extrude into the lumen [
From a scientific standpoint using a statistical treatment, the growth perfor- mance and morphometric parameters of intestinal villi have no alterations. When these values are expressed as an index of 100, the experimental groups showed a value of 104, particularly in the body weight gain of the 50 ppm DFG + 250 ppm FCP was 106. Within poultry production, a supplement resulting in a body weight gain of more than 2% than the control is considered effective. Regarding growth performance, although most of the percentages of the carcass, thigh, breast and wing in the experimental groups were higher than the control, the abdominal fat decreased, suggesting that the DFG and FCP affect the growth performance but suppresses fat deposition. Regarding the gross anatomical observations of the intestine and visceral organs, we could not find a difference among the groups. However, the villus height and area showed higher values in the transition from the ileum to the duodenum. This is in harmony with reports that the duodenum and initial jejunal segment absorbed lipids, while the ileum did not play a significant role in digestion or the absorption of triglycerides [
In conclusion, a dietary mixture of a small amount of DFG and a small amount of FCP induced protuberated epithelial cells and deeper cells at the sites of recently exfoliated cells on the intestinal villus apical surface in the duodenum and jejunum, suggesting an activated function of the cells. This cell activation induced a 14% increase in duodenal villus height and a 24% increase in jejunal villus area, resulting in a 6% increase in body weight gain. The present results suggest that a small amount of fermented corncob powder can be used as a feed supplement when mixed with fermented ginger powder, due to their synergistic interaction.
Khonyoung, D., Sittiya, J. and Yamauchi, K. (2017) Growth Performance, Carcass Quality, Visceral Or- gans and Intestinal Histology in Broilers Fed Dietary Dried Fermented Ginger and/ or Fermented Corncob Powder. Food and Nutrition Sciences, 8, 565-577. https://doi.org/10.4236/fns.2017.85039