Influence of different levels of humic acid and esterified glucomannan on growth performance and intestinal morphology of broiler chickens

doi:10.4236/as.2012.35080

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Vol.3, No.5, 663-668 (2012) Agricultural Sciences

http://dx.doi.org/10.4236/as.2012.35080

Influence of different levels of humic acid and

esterified glucomannan on growth performance

and intestinal morphology of broiler chickens

Seyed Mozafar S. M. Taklimi1, Hassan Ghahri2, Mohammad Asadi Isakan1

1Department of Animal Physiology and Nutrition, Animal Science Research Institute, Karaj, Iran;

*Corresponding Author: [email protected], [email protected]

2Urmia Branch Islamik Azad University, Urmia, Iran

Received 25 January 2012; revised 7 March 2012; accepted 2 May 2012

ABSTRACT

A s tudy was under taken to investigate the effe ct

of dietary supplementations of humic acid and

esterified glucomannan on broiler performance

and the morphological measurements of small

intestine. Two hundred and ten-day-old broiler

chicks were weighed and randomly assigned to

seven treatment groups in triplicate. The dietary

treatments included (T1) control (basal diet

without any additives), (T2) basal diets supple-

mented with 0.1% humic acid, (T3) basal diets

supplemented with 0.2% humic acid, (T4) basal

diets supplemented with 0.3% humic acid, (T5)

basal diets supplemented with 0.1% esterified

glucomannan, (T6) basal diets supplemented

with 0.2% esterified glucomannan and (T7) basal

diets supplemented with 0.3% esterified gluco-

mannan. Different parameters including body

weight gain (BWG), feed intake (FI), feed con-

version ratio (FCR) and morphology of small in-

testine were evaluated during six weeks of ex-

perimental period. According to the results, at

the end of experimental period, the highest av-

erage BWG were recorded in T4 group (fed with

0.3% humic acid), compared to control (T1) and

other experimental groups (p < 0.05). Whereas,

the highest body weight gain were observed

during starter period in treatment 7 (fed with

0.3% esterified glucomannan) as compare to

other experiment al groups (p < 0.05). S t atistically,

feed intake remained unaffected during the ex-

perimental period in all the treatment groups.

However, the lowest and the highest feed intake

were observed numerically within treatments T4

and T1 with 4229.70 g and 4362.30 g, respec-

tively. Addition of dietary supplements used in

the study appeared to have significant effect on

the morphology of the small intestine (jejunum

mucosal development) of the broilers in differ-

ent tre a t m e n t g r o u ps. C om par ed t o c o n tr o l group,

the inclusion of either humic acid or esterified

glucomannon decreased (p < 0.01) the crypt

depth and increased villus height respectively

(p < 0.05). Moreover, the diet supplements with

humic acid 0.3% (T4) decreased crypt depth

compared to esterified glucomannan and con-

trol. The dietary supplementations resulted in an

increase in the villus height of intestinal mucosa

of broilers. The increase in the villus height was

associated with improvement of growth perfor-

mance for both humic acid and esterified glu-

comannan. Based on our result s it appeared that

humic acid and esterified glucomannan can be

used as a growth promoter in broiler diets and

they can improve the gut health too.

Keywords: Broiler Chicks; Humic Acid; Esterified

Glucomannan; Small Intestine

1. INTRODUCTION

Any attribute which can improve gastrointestinal tract

health and immunity is extremely valuable in broiler

diets. In last decades antibiotics had been used widely

world-wide in poultry industry in order to prevent poul-

try pathogens and disease and to improve meat and egg

production. However, with the unavoidable spread of

bacterial resistance and cross resistance the use of antibi-

otics has been considered hazardous [1-3]. As conse-

quences, many alternatives have been discussed includ-

ing probiotics, prebiotics, herbal compounds and organic

acids. Most of these substances exert their effects by in-

fluencing gastrointestinal flora and digestion processes.

Among these alternatives the influence of humic acid has

been studied extensively, and has been reported to have

S. M. S. M. Taklimi et al. / Agricultural Sciences 3 (2012) 663-668

664

positive impact on the growth performance of birds.

Humic acid defined as a class of compounds resulting

from decomposition of organic matter, particularly plants

are natural constituents of drinking water, soil and lignite.

Humic acid inhibit bacterial and fungal growth, thus de-

crease levels of mycotoxins in feed [4,5]. Beneficial ef-

fects of humic acid are described concerning stress ma-

nagement [6], immune system [7,8], anti-inflammatory

activity [9], antiviral properties [10], as well as preven-

tion of intestinal diseases, mainly diarrhea in humans and

animals. There are also reports that routine use of humic

acid in the feed has a positive influence on growth of

broilers [11,12]. The beneficial effect of humic acid on

growth in different species of animals has been consid-

ered its capacity of changing gut physiology and inter-

ference in immunity [13]. Beside humic acid, yeast and

yeast cultures have a long standing tradition of use in

animal feeds. Yeast culture has been shown to affect in-

testinal mucosa development in poultry. In addition to

growth performance, there are many trials showing that

enrichment of diets with yeast could favorably improve

the quality of edible meat from broilers. Yeast is also an

excellent source of selenium and chromium; two trace

minerals which may have positive effects of broiler

health [14]. Since, 80’s decades Oligosaccharide from

outer cell wall component of live yeast (Saccharomyces

cerevisiae), were introduced as a feed additive commer-

cially in poultry industry. Since then, many studies have

been conducted to investigate the effects of esterified

glucomannan or it’s derivatives on various parameters

such as body weight, feed conversion ratio and feed in-

take of birds [15-17]. The objective of this study was to

determine if experimental supplementation of different

level of humic acid and esterified glucomannan in the

young broiler chicken diet may improve performance

and small intestine mucosal development of broiler

chicks.

2. MATERIALS AND METHOD

2.1. Birds and Experimental Design

Two hundred and ten-day old (mixed sex) commercial

Ross broiler chicks were randomly subjected to seven

different treatment groups (three replicates of 10 chicks

per dietary treatment). All birds were weighed (initial

weight) individually after their arrival from the hatchery

to the experimental farm, and on completion of the ex-

periment day 42. The experiment was designed com-

pletely randomized to evaluate the effect of two different

naturally available feed additives humic acid (FH =

Farmagu lator D RYTM humate Yayalar Mah. Sanayi Cad.

No. 29, Dolayoba, Pendik, Istanbul, Tu rkey ), and esteri-

fied glucomannan, (Bio-Mos, USA) on small intestine

mucosal development of digestive tract of birds. The

birds were fed one of seven experimental treatments for

42 days. The experimental diets were as follows: T1 =

Control (basal diet + no feed additives). T2 = T1 + 0.1%,

T3 = T1+ 0.2% and T4 = T1+ 0.3% humic acid and T5 =

T1 + 0.1%, T6 = T1 + 0.2% and T7 = T1 + 0.3% esteri-

fied glucomannan, respectively.

2.2. Performance

All chicks were reared under uniform management

conditions with feed and water available ad libitum. In-

dividual body weight gain (BWG), feed intake (FI), feed

conversion ratio (FCR) of the birds were recorded at the

onset of experiments and measured daily thereafter for

each group.

2.3. Small Intestine Mucosal Development

Studies

To study the mucosal development of small intestine

of the chicks at the end of experimental period, 3 birds

were selected randomly from each replicate and sacri-

ficed and their histomorphology studied.

Gut morphology (jejunum) examinations were carried

out according to the method which was described [18].

Jejunum samples were immersed in 4% formalin for 48

hrs. The processing consisted of serial dehydration, clear-

ing, and impregnation with wax. Tissue sample was sec-

tioned at a thickness of 7 - 8 μm, were cut by a micro-

tome and were fixed on slides. A routine staining proce-

dure was carried out using hematoxylin and eosin. The

slides were viewed on a microscope. Visual observations

of villus height, crypt depth were made at 10.25× magni-

fication under an IMI microscope.

2.4. Statistics

All data were statistically analyzed by ANOVA using

the SAS system [19]. Significant differences between the

treatment means were compared by using Duncan Multi-

ple Range test [20].

The statistical model used was

Yij = µ + Ti + eij

where,

Yij = observation in block i and treatment j, µ = Over-

all sample mean, Ti = Effect of treatment i, eij = Error.

3. RESULTS

The effect of different levels of humic acid and esteri-

fied glucomannan on body weight gain of broilers during

the experimental period is shown in Table 1. A sig-

nificant increase in BWG was recorded in birds which

was fed with either humic acid or esterified glucomannan

(p < 0.05). According to the results, birds fed with 0.3%

S. M. S. M. Taklimi et al. / Agricultural Sciences 3 (2012) 663-668 665

Table 1. Effects of humic acid and esterified glucomannan on

mean BWG (g) of Broiler.

0 - 42 days21 - 42 days 0 - 21 days Treatments

2147.00b 1459.67b 687.33c T1 (control)

2267.33ab

1554.67bc 712.67bc T2 (0.1% Humic acid)

2337.67a 1611.33a 726.33abc T3 (0.2% Humic acid)

2373.67a 1650.33a 723.33abc T4 (0.3% Humic acid)

2277.67ab

1551.00ab 726.67abc T5 (0.1% Live yeast)

2363.33a 1622.67a 740.67ab T6 (0.2% Live yeast)

2331.67a 1602.00a 763.00a T7 (0.3% Live yeast)

* * *

*Mean bears not common superscript are different significantly (p < 0.05).

a,b,cmeans: Values in the table shows with different superscripts are signifi-

cantly different (p < 0.05).

estrified glucomannan performed well at the early age,

while with advancing age, humic acid had an increasing

effect on overall performance of the birds. The highest

and the lowest BWG was observed in T7 and T1 (763.00

vs 687.33 g) during their initial growing phase (0 - 21

days), respectively, while, chicks in T4 (0.3% humic acid)

gained more body weight during the 2nd phase of growth

(21 - 42 days) as well during the whole experimental

period (p < 0.05).

As Table 2 indicates, feed intakes were not affected

significantly by inclusions of different levels of either

humic acid or esterified glucomannan in diets, during the

experimental period. However, feed conversion ratio of

the chicks in different treatment groups were signifi-

cantly different with the presence of either humic acid or

esterified glucomannan. According to the results pre-

sented in Ta b l e 3, the highest feed conversion ratio was

observed in T4 (1.781), as compared to other experimen-

tal groups (p < 0.01). Overall results indicated that, birds

fed diets with humic acid (0.3%) had higher mean live

weight gain, low feed intake and better feed conversion

ratio.

Table 4 indicated that, feed additives used in this

study had significantly effect on the villus height (p <

0.05) of broiler chicks (Figure 1). All levels of humic

acid and esterified glucomannan used had the ability to

increase crypt depth (p < 0.01). Where as, highest crypt

depth were observed in T7, compared to other treatments.

As evidence shows that, effects of humic acid and esteri-

fied glucomannan, is proposed to be due to their contents

of anti bacterial agents and enhancement of multiplica-

tions of beneficial macroscopic bacteria and elimination

of harmful bacteria in bird's digestive tract.

4. DISCUSSION

According to the results of this experiment, humic acid

Table 2. Effects of humic acid and esterified glucomannan on

FI (g) of Broiler.

0 - 42 days21 - 42 days 0 - 21 days Treatments

4362.30 3274.30 1088.00 T1 (control)

4295.00 3217.30 1077.67 T2 (0.1% Humic acid)

4285.70 3253.00 1032.67 T3 (0.2% Humic acid)

4229.70 3172.00 1057.67 T4 (0.3% Humic acid)

4274.30 3217.30 1042.00 T5 (0.1% Live yeast)

4317.00 3257.00 1060.00 T6 (0.2% Live yeast)

4327.70 3265.70 1062.00 T7 (0.3% Live yeast)

ns ns ns

ns = Not significantly different (p < 0.05).

Table 3. Effects of humic acid and esterified glucomannan on

mean FCR.

0 - 42 days21 - 42 days 0 - 21 days Treatments

2.031a 2.243a 1.583a T1 (control)

1.894b 2.069b 1.512ab T2 (0.1% Humic acid)

1.833c 2.018bc 1.421b T3 (0.2% Humic acid)

1.781d 1.922c 1.462ab T4 (0.3% Humic acid)

1.876c 2.074b 1.433b T5 (0.1% Live yeast)

1.826c 2.00bc 1.431b T6 (0.2% Live yeast)

1.856c 2.038b 1.392ab T7 (0.3% Live yeast)

** *

*Mean bears not common superscript are different significantly (p < 0.05);

**Mean bears not common superscript are different significantly (p < 0.01).

a,b,cmeans: Values in the table shows with different superscripts are signifi-

cantly different (p < 0.05).

Table 4. Effects of humic acid and esterified glucomannan on

Jejunum mucosal development of broiler.

Crypt depth (mm)Length of villiae (mm) Treatments

2.243a 1.026b T1 (control)

2.069b 1.290ab T2 (0.1% Humic acid)

2.018bc 1.383a T3 (0.2% Humic acid)

1.922c 1.340a T4 (0.3% Humic acid)

2.074b 1.260ab T5 (0.1% Live yeast)

2.00bc 1.293ab T6 (0.2% Live yeast)

2.038b 1.246b T7 (0.3% Live yeast)

** *

*Mean bears not common superscript are different significantly (p < 0.05);

**Mean bears not common superscript are different significantly (p < 0.01).

a,b,cmeans: Values in the table shows with different superscripts are signifi-

cantly different (p < 0.05).

S. M. S. M. Taklimi et al. / Agricultural Sciences 3 (2012) 663-668

666

Figure 1. Vertical view of small intestine (jejunum, depth of crypt and length of villae, 100× magnifications).

and esterified glucomannan added as supplement in the

feed had significantly effect on the performance of birds.

This is in general agreement with a number of recent

reports [14,12,21,22]. Nevertheless, Humic acid had

higher effects on birds as compare to esterified gluco-

mannan by means of 1) Ability to create protective layers

over the epithelial mucosal membrane of digestive tract

against the penetrations of toxic and other bacterial con-

taminated substances; 2) Will have ability to increase

metabolism of protein and microbial carbohydrates based

on catalyzing agents by bringing down the pH of small

intestine; 3) It acts as detoxification agents in intestine

(due to reducing power in absorption of nitrates, fluorites

and heavy metals); 4) To increase immune receptors in

digestive tract over preserving the beneficial pathogens

[23]. There is some evidence that shows, by inclusions of

esterified glucomannan in feed will increase bird’s

growth through reducing the harmful gut micro phlora by

de-colonizing salmonella like bacteria, will increase

bird’s body weight, immune system, nutrient digestibility

and gut health due to its effect on substituting harmful

with beneficial gut micro phlora [17]. Esterified gluco-

mannan will also shows its beneficial effect on birds

through challenge with some un-expected growth inhibi-

tors like aflatoxin, coccodiosis, heat stress and poor feed

quality [17]. Feed intake was also affected numerically

by inclusions of humic acid or esterified glucomannan at

different levels throughout the experimental periods.

Birds had offered diets containing 0.3% humic acid had

lower levels of feed intake as compare to other experi-

mental groups. This finding was in accordance with the

reports of [21,22]. Feed conversion ratio was affected by

inclusions of humic acid and esterified glucomannan

throughout the experimental period. Best feed conversion

ratio was observed in 0.3% humic acid supplemented

groups, and it was in accordance with the reports of

[12,14]. It is assumed that, due to the chemical composi-

tions like, proteins, water soluble vitamins, digestive

enzyme and many other immune stimulating agent and

antibacterial substances in humic acid and in esterified

glucomannan, they will have significant role in the of

health and productivity of birds. This could be mainly

attributed to their ability to change the gut micro flora

(by increasing the concentrations of beneficial bacteria)

in the intestine [24-26]. In the present study, inclusion of

0.3% humic acid in broiler diets had significantly better

effects on bird performance. As reported earlier by

[27,28], humic acid and esterified glucomannan had sig-

nificant effect on crypt depth of villiae in Jejunum of

treated birds compared to non supplemented groups. It is

obviously known that, growth of villiae will be depend-

ing on pH, micro flora and toxic substances in the intes-

OPEN ACCESS

S. M. S. M. Taklimi et al. / Agricultural Sciences 3 (2012) 663-668 667

tine. Though, humic acid will have the ability in reducing

pH and concentration of harmful bacteria in intestine.

There is evidence that humic acid could have positive

effect on animal performances via digestive tract ecosys-

tems [29]. Esterified glucomannan at higher dosage will

increase height of villiae and crypt depth and inhalations

of damaged villiae [30]. It also will secrete more en-

zymes in taller (projected) villiae, which finally will en-

hance the absorption and digestion of food particles.

Moreover, esterified glucomannan will have effect to

create protective layers on intestinal mucus, to inhibit

penetrations of toxic substances and harmful bacteria in

the gut.

To conclude, humic acid or esterified glucomannan

displayed a greater efficacy as growth promoters for broil-

ers and could be a suitable substitution as an alternative

for antibiotics.

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