International Journal of Applied Research and Technology 29

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International Journal of Applied Research and Technology

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Residual Antinutrients in Differently Processed Jatropha Curcas Kernel Meals: Effect on Blood Parameters and Gut

Microbes of Broiler Chicks

Ojediran, T. K., Olayeni, T. B., Shittu, M. D., Ogunwemimo, O. T. and Emiola, I. A.

Ladoke Akintola University of Technology, Ogbomoso, Nigeria.

Available online: January 30, 2015

To cite this article:

Ojediran, T. K., Olayeni, T. B., Shittu, M. D., Ogunwemimo, O. T. and Emiola, I. A. (2015). Residual Antinutrients in

Differently Processed Jatropha Curcas Kernel Meals: Effect on Blood Parameters and Gut Microbes of Broiler Chicks.

International Journal of Applied Research and Technology. 4(1): 29 – 38.

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International Journal of Applied Research and Technology 30

International Journal of Applied Research and Technology Esxon Publishers

Vol. 4, No. 1, January 2015. 29 – 38.

Residual Antinutrients in Differently Processed Jatropha Curcas Kernel

Meals: Effect on Blood Parameters and Gut Microbes of Broiler Chicks

Ojediran, T. K., Olayeni, T. B., Shittu, M. D., Ogunwemimo, O. T. and Emiola, I. A.

Ladoke Akintola University of Technology, Ogbomoso, Nigeria.

(Received: 18 December 2014 | Accepted: 30 December 2014 | Published: 30 January 2015)

Abstract

This study was conducted to determine the effects of differently processed Jatropha curcas kernel meal diets on the blood

profile and gut microbes of broiler chicks. Raw Defatted Meal (RDM), Toasted Defatted Meal (TDM), Cooked Defatted

Meal (CDM), Lye Defatted Meal (LDM) and Sand Roasted Defatted Meal (ZRDM) Jatropha curcas kernel meals were each

included at one-third replacement for soya bean meal in a feeding trial that lasted for 21 days. 180 day-old Marshal strain

unsexed broiler chicks were used for this experiment. There were 6 dietary treatments (control inclusive) of 30 birds per

treatment and 3 replicate of 10 birds each in a complete randomized design. The result suggests that heat treatments reduced

the antinutrients with minimal effect on the saponin and phorbol esters present in the Jatropha curcas kernel meals. All

chicks on the dietary treatments had most of the serum parameters affected: elevated globulin levels signifies liver and kidney

damage which can be attributed to the residual antinutrients in the meals, most especially phorbol esters. The dietary

treatment confers some antimicrobial properties, especially against E. coli which is indictive of the residual antinutrients..

However, further experiments should be carried out to examine the performance of broiler chicks on bio-treated samples of

the processed Jatropha curcas kernel meals.

Keywords: Antinutrients, Blood, Broilers, Gut Microbes, Jatropha curcas Kernel, Processing.

For corresponding author:

E-mail: info@esxpublishers.com

Subject: 0214-0211

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International Journal of Applied Research and Technology 31

Introduction

Research into alternatives and cheaper sources of feedstuffs or ingredients to the orthodox feedstuffs in order to widen

sources of raw materials for poultry feeds (Annongu et al., 2010) has been exploited in recent years, but, most of these

alternatives come with the challenges of antinutrients (Emiola et al., 2003, 2004, 2007; Belewu et al., 2010; Akande et al.,

2010, 2014). The use of Jatropha curcas meal in animal nutrition is however faced with several problems of anti-nutritional

factors such as lectin, saponin, tannin, phytate, trypsin inhibitors and phorbol esters (Makkar and Becker, 1999). Due to these

phytotoxins, the seeds or cakes or its oil cannot be used for human or animal consumption but, the seed has about 35-50%

crude protein (Aslani et al. 2007), 60% oil and rich in essential amino acid and minerals (Makkar et al. 2008) is attractive to

feed researchers. Jatropha curcas also has been reported to possess antimicrobial properties (Oyi et al., 2007). The decreases

in the levels of anti-nutritional factors to safe limits may be caused by thermal degradation, soaking in distilled water,

germination, and extraction of methanol (Yasmin et al. 2008; Magdi, 2007; Aderibigbe et al. 1997), hot water treatment, lye

treatment and fermentation (Akande et al. 2012). Nevertheless, blood parameters are a good diagnostic tool to examine the

influence of feedstuffs or antinutrients on the physiological well-being of livestocks.

This study therefore attempts to investigate the effect of residual antinutrients in differently processed Jatropha

curcas kernel meals on the blood parameters and gut microbes of broiler chicks.

Materials and Methods

The research was conducted at the poultry unit of the Teaching and Research Farm, Ladoke Akintola University of

Technology, Ogbomoso. Dry seeds of J. curcas were purchased locally. The seeds were dehulled manually to separate the

kernel from the shell. The extraction of oil follows similar manner to that of other oilseeds such as cashew nut meal, castor

seed cake (Odunsi et al. 2002; Akande et al. 2012). The kernels were divided into portions for ease of processing. Five

different processing methods these include; A portion of the kernel was milled and subjected to oil extraction using hydraulic

press and was referred to as Raw Defatted Meal (RDM) (Akande et al., 2012). A potion of the milled kernel from was

roasted until it becomes crispy to touch and turn brown in a pan. It was stirred from time to time to maintain uniform heating

while it lasted for 30 minutes, as this was referred to as Toasted Defatted Meal (TDM). A portion of the raw kernel was

cooked at 1200C ±50C for 30 minutes (similar to the procedure of Martinez-Herrera et al., (2006)) in a cooking pot, sun

dried for 24 hours, after which they were oven dried at 850C for an hour before being milled and then defatted using the

hydraulic press as this was referred to as Cooked Defatted Meal (CDM). The lye was prepared by putting wood ash in a

muslin cloth and hot water (1000C ± 50C) was poured on the ash and the filtrate (pH 9.5) was used to cook the kernel at

1200C ± 50C and held for 30 minutes. This is a variant of the procedure of Akande, (2010). The treated kernel was dried,

milled and defatted and referred to as Lye Defatted Meal (LDM). Raw whole seeds were roasted in sand (particle size of ¼–

½ mm ) at 115oC ±50Cand held at this temperature for 30 minutes. The roasted seed was cooled, dehulled and kernels were

milled then defatted to produce Sand Roasted-Defatted kernel Meal (ZRDM). All meals were at between 0.5-1.0mm mesh

size.

Six (6) experimental diets were formulated: Diets 1 contained 0% JKM and served as the control diet, while diets

2, 3, 4, 5 and 6 contained 10.33% (one-third replacement of soybean meal) inclusion level of RDM, TDM, CDM, LDM and

ZRDM respectively as shown in table 1. All diets were iso-nitrogeneous and iso-caloric. One hundred and eighty (180)

Marshal Strain Broiler Chicks were used for this study. All the birds were initially fed on commercial broiler starter mash

for the first week to stabilize the chicks after which they were randomly distributed without sexing into six dietary groups of

thirty (30) birds each. Each treatment group was further sub-divided into three replicates of ten (10) birds each. The birds

were fed with their respective treatment diet and water was served ad-libitum. The experimental chicks were raised under

intensive care management in a deep litter system. Occasional management practices such as vaccination, medication, weekly

weighing of birds and feed intake, changing of litters and proper record keeping were taken. The study lasted for 21 days for

the introduction of dietary treatments.

Trypsin inhibitor was determined using the method of Kakade et al. (1969); Lectin content was determined by

hemaglutination assay as described by Makkar et al. (1997); Tannins was determined using the method of Swain (1979);

The determinations of total saponins were applied using a spectrophotometric method described by Hiai et al. (1989); Phytate

was determined using the method of Maga, (1983) while Phorbol esters was determined after the procedure of Haas and

Mittelbach, (2000). Three birds per treatment were randomly selected and slaughtered by cutting the jugular vein. About 5ml

of blood was collected into two sets of three sterilized glass bottles/tubes. For haematology, blood samples were collected

into two sets of three sterilized bottles containing Ethylene Diamine Tetra-acetic Acid (EDTA). Blood samples for serum

biochemical studies were collected in plain bottles (i.e. without anticoagulant) for serum separation. Serum was obtained by

centrifugation and serum samples were stored in a deep freezer (at minus 100C) until required for analysis.

Blood parameters such as packed cell volume (PCV) and haemoglobin (Hb) were determined using the micro

haematocrit method and cyanmethaemoglobin methods respectively as described by Mitruka and Rawnsley (1977).

Erythrocyte count (RBC) and Leukocyte count (WBC) were determined using the improved Neubauer haemocytometer after

the appropriate dilution (Schalm et al., 1975). Differential leukocyte counts were determined by scanning Giemsa’s stained

slides in the classic manner (Schalm et al., 1975).

Mean corpuscular haemoglobin (MCH), Mean corpuscular haemoglobin concentration (MCHC) and Mean

corpuscular volume (MCV) were calculated using the following formula:

MCH = Haemoglobin × 10

RBC

International Journal of Applied Research and Technology 32

MCHC= Haemoglobin x 100

Hematocrit

MCV = Hematocrit x 10

RBC

Glucose and cholesterol were determined by spectrophotometric methods. Alanine Transaminase (ALT), Aspartate

Transaminase (AST) and Alanine phosphatase (ALP) was determined manually by spectrophotometric method respectively

as described by Schmidt (1963). Urea was determined by urease method and creatinine by Folin-Wu filtrate methods. Total

serum protein was determined using the biuret method as described by Reinold (1953) while albumin was determined using

the BCG (Bromocresol green) method as described by Peters et al. (1982).Mann Rogosa Sharpe (MRS) agar was used for

colony count of Lactobacillus growth. These were prepared according to the methods recommended by Harrigan, (1998).

The pH of media was adjusted by using 0.1N NaOH and 0.1N HCl. The composition of various media is given in Table 1.

This was the selective medium used for the isolation and enumeration of Lactobacillus spp. The ingredients were

dissolved in1000mi distilled water and the pH was adjusted at 6.2 to 6.6 and then the medium was sterilized at 1210C for

15minutes under 15lb pressure. MRS agar was prepared by adding 1.5% agar in MRS broth (prepared above) and was then

autoclaved. A suspension of 100ml was made from 1g of the sample mixed with 99ml of the given sample which after

shaking at 250 rpm for 20 minutes. Serial dilution (10-2 to 10-5) were made in 0.9% NaCl solution and then spread in plates

in duplicate. After incubation at 320C for 24hr, ten colonies were randomly picked from the MRS agar plates. 51.5g of

powder was weighed and 1litre of deionised water was added, allowed to soak for 10minutes, swirl to mix, then sterilized

for 15 minutes at 1210C then cooled to 470C and pour into petri dishes (Table 2). Glass wares as Conical flasks, measuring

cylinder were washed throughly with detergent, rinse with water and sterilized in the oven at 1400C for 180 minutes. They

were then wrapped properly with aluminium foil before sterilization (Fawole and Oso, 2001).

All data generated and estimated (for blood parameters, others were a means of triplicates) were subjected to

Analysis of Variance in a Complete Randomized Design of SAS (2000) software package. Significant means were separated

using a Duncan multiple range test of the same package.

Results and Discussion

The results of the effects of different processing methods on anti-nutritional components of Jatropha curcas kernel meals

are presented in Table 4. The Raw kernel (RWK) had the highest content of antinutrients followed by the Raw defatted meal

(RDM). Heat treatments reduced the contents of trypsin inhibitor in Toasted defatted meal (TDM), Cooked defatted meal

(CDM), Lye defatted meal (LDM) and Sand-roasted defatted meal (ZRDM). Lectin, Tannin, Saponin, Phytate and

phorbolesters content of RWK was higher than the defatted meals while TDM had none for lectin and least for tannin,

saponin, phytate and phorbolesters. As shown in Table 5. Pack cell volume (PCV), Haemoglobin (Hb), White blood cell

(WBC), Red blood cell (RBC), Platelet, Mean corpuscular volume (MCV), Mean corpuscular haemoglobin (MCH) and

Mean corpuscular haemoglobin concentration (MCHC) were affected (p<0.05) by the dietary treatments. Values obtained

for PCV, Hb, WBC and RBC among birds fed processed Jatropha curcas kernel meals (JCKM) were higher (p<0.05) than

those fed the control with the exception of those fed diet 5. Meanwhile, WBC, MCH and MCHC followed a similar trend

(p<0.05).

Table 6. showed the serum parameters of broiler chicks fed differently processed Jatropha curcas kernel meals.

Total Protein (TP), Globulin (GLOB), Albumin (ALB), Aspartate Transaminase (AST), Cholesterol (CHOL), Triglyceride

(TG) and Creatinine were significantly affected (p<0.05) and an inconsistent pattern was observed. Effect of treated Jatropha

kernel meals on microbial population in the GIT of broiler chicks is shown in table 7. It was evident that the different

processed JKC contain antimicrobial properties that reacted against both gram positive and negative stain bacteria.It was

observed that Diets 2, 3, 4, 5 and 6 contained antimicrobial properties that reacted against the population of E-coli compared

to Lactobacillus. Diet 1 (control) had Lactobacillus and E. coli populations unlike birds fed Diets 2, 3, 4, 5 and 6. The

reduction in TIA confirms the fact that trypsin leached out with processing and are heat labile (Siddhuraju et al., 1996),

though, some may be resistant at this temperature as reported by Magdi (2007). Also, no amount of TIA was observed in the

TDM and could be due to the temperature during toasting. Meanwhile, Jyothi and Sumathi (1995) reported that the extraction

at both low and high temperatures with sodium bicarbonate was most effective in the case of trypsin inhibitors of common

bean seeds. Heat labile nature of lectin is contrary to an earlier study by Aderibigbe et al. (1997) who found an increase in

lectin activity following heat treatment which was attributed to some artifacts.

Reddy and Pierson (1994) reported that saponins are not destroyed by cooking. Also, Abou Arab and Abu-Salem

(2010) observed no significant difference in saponin contents of defatted whole seed and kernel after roasting. The phytate

content of Jatropha meals was much higher than that of peanut presscake (1.36%) (Fardiaz and Markakis, 1981). These

values suggest presence of high levels of phytate in Jatropha curcas samples. These high levels of phytate might decrease

bioavailability of minerals. Phytates have also been implicated in decreasing protein digestibility by forming complexes and

also by interacting with enzymes such as trypsin and pepsin (Reddy and Pierson, 1994). Aderibigbe et al (1997) observed

that none of the heat treatments studied decreased phytate level. Belewu and Sam (2010) observed 0.013 phorbol ester for

RDM, meanwhile, it is noteworthy that, the content of phorbolester in most of the samples was still high. Phorbolester content

International Journal of Applied Research and Technology 33

of 0.09 mg/g in seed meal is considered safe for livestock because the content is lower than that found in non-toxic jatropha

cultivar (0.11 mg/g) (Makkar et al. 1998). These findings are in conformity with the work of Belewu (2008) who reported

the death of albino rats fed fungus treated Jatropha seed meal.

Heamatological examination is an indirect assessment of the clinical and nutritional health status of animals. The

commonly examined heamatological parameters in nutritional studies include PCV, RBC, Hb, MCHC, MCV and clotting

time (Aletor and Egberongbe, 1992; Olorode and Longe, 2000; Adeyemi et al., 2000). The PCV values obtained in this study

were within the range reported by Tambuwal et al. (2002). This could be due to Compensatory Accelerated Production

(CAP) of packed cell volume which returns the PCV levels to normal level (Ganong, 2001; Tambuwal et al., 2002). However,

the value obtained for PCV and RBC were similar to those reported for chicks fed winged bean and full fat jatropha seeds

(Igene, 1990; Adeyemi et al., 2000). The PCV, Hb, WBC, RBC values obtained in this study were within the normal range

described by Mitruka and Rawnsley (1977) while Platelet, MCV, MCH and MCHC were not. Birds fed the control diet,

RDM, LDM and ZRDM were within the range while the TDM and CDM were outside the range. The significantly higher

values of white blood cell recorded in diets containing RDM, TDM, and CDM could be as a result of the birds possessing a

protective system suggestive of a well adapted immune system. Similarly, a deficiency in any type of normal white blood

cell may result in an increased susceptibility to infections. The values of RBC on all the diets are comparable to those reported

by Mmereole, (2008).

It has been reported that serum biochemical constituents positively correlate with the quality of the diet (Adeyemi

et al., 2000). ALT, ALP and ACP were not affected (p>0.05). Total serum protein is influenced by breed, age, physiological

state, environmental and antigen exposure and levels can be extremely variable (Bell, 1971). Globulin levels may be elevated

in chronic infections, liver damage, leukemia and kidney dysfunction and decreased in nephrosis (a condition in which the

kidney does not filter the protein from the blood and it leaks into the urine), acute hemolytic anaemia and liver dysfunction

(Sanchez-Monge et al., 2004). Serum albumin will increase when protein intake exceeds the amount required for growth and

maintenance. The values obtained for AST were not within the normal range (88.0-208U/L) as described by Mitruka and

Rawnsley, (1977), while globulin, ALT, creatinine were within that range. The main cause of serum ALP reduction is a

damage of the intestine (Rivets et al., 1975). Appearance of abnormal amounts of certain enzymes of intercellular origin in

the blood reflects damage to an organ or tissue (Wilson, 2008). The liver is rich in some enzymes as ALT, AST and its

damage often results in releasing these enzymes in the blood. Akande et al. (2014) noted that the creatinine may reduce since

toxins gets to the liver before the kidney.

The anti-microbial properties in the latex of Jatropha curcas are against both gram positive (lactobacillus) and gram

negative (E. coli) bacteria is supported by Oyi et al. (2007). Antibiotic activity of Jatropha has been observed against

organisms including Staphylococcus aureus and Escherichia coli (Thomas et al., 2008). The antimicrobial activities of the

latex could be due to the presence of secondary metabolites such as tannins, flavonoids and saponins which have been

confirmed to be present. The kernel meal methanol extract showed good antibacterial activity, although it contained low

amounts of terpenes, flavonoids and phenolics. Its activity may have come from other antimicrobial agents such as saponins

or phorbol esters that were observed to be present in the kernel meal (Namuli et al., 2011).

Conclusion and Recommendations

Bird fed diets containing LDM were anaemic while all chicks on the dietary treatments had most of the serum parameters

affected: elevated globulin levels signifies liver and kidney damage and these can be attributed to the residual antinutrients

in the meals, most especially phorbol esters. The dietary treatment confers some antimicrobial properties especially against

E. coli which is indictive of the residual antinutrients.

Acknowledgement

We would like to appreciate the Department of Animal Nutrition and Biotechnology, Ladoke Akintola University of

Technology, Ogbomoso, Nigeria, under the headship of Prof Wale Emiola for making the facilities used during the course

of these experiment available.

International Journal of Applied Research and Technology 34

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International Journal of Applied Research and Technology 36

Tables

Table 1: Composition of MRS Agar

Ingredients Quantity (g)

Peptone 10.00

Meat extract 10.00

Yeast extract 5.00

D-glucose 20.00

Tween 80 1.00

K2HPO4 2.00

Sodium acetate 5.00

Tri-ammonium citrate 2.00

MgSO4.7H20 0.20

MnSO4.4H20 0.05

Table 2: Composition of MacConkey Agar no. 3

Formula g/litre

Peptone 20.00

Lactose 10.00

Bile Salt no. 3 1.50

Sodium chloride 5.00

Neutral red 0.03

Crystal violet 0.001

Agar no. 2 15.00

Table 3: Gross Composition of Experimental Diets for the Broiler Starters

Ingredients % Diet 1

(CNRL)

Diet 2

(RDM)

Diet 3

(TDM)

Diet 4

(CDM)

Diet 5

(LDM)

Diet 6

(ZRDM)

Maize 53.00 53.00 52.00 53.00 53.00 52.00

Wheat offal 6.00 6.00 7.00 6.00 6.00 7.00

Soybean meal 31.00 20.67 20.67 20.67 20.67 20.67

JKM 0.00 10.33 10.33 10.33 10.33 10.33

Fish meal 6.00 6.00 6.00 6.00 6.00 6.00

Limestone 1.35 1.35 1.35 1.35 1.35 1.35

DCP 2.00 2.00 2.00 2.00 2.00 2.00

Salt 0.20 0.20 0.20 0.20 0.20 0.20

Vitamin Premix 0.25 0.25 0.25 0.25 0.25 0.25

Methionine

Lysine

0.15

0.05

0.15

0.05

0.15

0.05

0.15

0.05

0.15

0.05

0.15

0.05

Total (%) 100.00 100.00 100.00 100.00 100.00 100.00

Calculated Analysis

Crude protein (%) 23.66 23.31 23.43 23.17 23.91 23.61

M.E. kcal/kg 2940.82 3106.31 3100.57 3106.31 3106.31 3100.57

JKM= Jatropha curcas kernel cake meal, DCP= dicalcium phosphate

*Vitamin premix contained the following vitamins and minerals in 1kg of broiler diet: 12500 IU Vit. A; 2500 IU Vit. D3; 40mg Vit.E;

2mg Vit.K3; 30mg Vit B1; 55mg Vit.B2;550mg Niacin; 115mg Calcium Pantothenate; 50mg Vit B6 ; 0.25mg Vit B12 ; 500mg Choline

chloride; 10mg Folic acid; 0.08mg Biotin; 120mg Manganese; 1000mg Fe; 80mg Zn; 8.5mg Cu; 1.5mg I; 0.3mg Co; 0.12mg Se and

120mg Antioxidant.

International Journal of Applied Research and Technology 37

Table 4: Effect of various processing methods on anti-nutritional composition of Jatropha curcas kernel and meals

Anti-nutrients RWK RDM TDM CDM LDM ZRDM

Trypsin inhibitor (TIU/mg) 22.69 13.52 0.00 0.65 0.47 0.53

Lectin (HU/mg) 48.29 31.31 0.00 4.23 2.55 3.18

Tannin(%) 0.070 0.053 0.007 0.019 0.014 0.016

Saponin (%) 2.18 2.09 1.24 1.74 1.53 1.61

Phytate (%) 8.63 8.26 1.84 2.46 2.05 2.39

Phorbolester(mg/100g) 2.700 2.490 1.040 1.287 1.140 1.207

%- percentage, RWK-Raw Whole kernel, RDM-Raw Defatted Meal, TDM-Toasted Defatted Meal, CDM-Cooked Defatted Meal, LDM-

Lye solution treated Defatted Meal, ZRDM-Sand Roasted Defatted Meal

Table 5: Heamatological parameters of broiler chicks fed differently treated jatropha curcas kernel meals

Parameters Diet 1 Diet 2 Diet 3 Diet 4 Diet 5 Diet 6 SEM

PCV(%) 31.95ab 39.00ab 41.80a 42.15a 28.95b 34.10ab ±1.63

Hb(g/dL) 9.25ab 11.8a 12.75a 13.05a 6.00b 10.45ab ±0.79

WBC(×103/µL) 225.65a 255.65a 263.80a 263.35a 128.45b 243.35a ±15.68

RBC(×106/µL) 2.14bc 2.81ab 2.99a 2.87ab 1.94c 2.42abc ±0.12

PLATELET(×103/µL) 14.00a 6.50b 10.00ab 6.50b 3.50b 5.50b ±1.08

MCV(FL) 150.00a 139.10b 140.15b 147.10a 149.80a 141.10b ±1.22

MCH(Pg) 43.20a 42.05a 42.70a 45.50a 22.30b 43.10a ±2.63

MCHC(g/dL) 28.85a 30.25a 30.50a 30.95a 15.05b 30.55a ±1.85 a, b, c, d = means within the same row bearing different superscripts differ significantly. PCV: Packed Cell Volume. Hb: Heamoglobin.

WBC: White Blood Cell. RBC: Red Blood Cell. MCV- Mean corpuscular volume. MCH- Mean corpuscular haemoglobin. MCHC- Mean

corpuscular haemoglobin concentration

Table 6. Serum parameters of broiler chicks fed differently processed jatropha curcas kernel meals

PARAMETERS Diet 1 Diet 2 Diet 3 Diet 4 Diet 5 Diet 6 SEM

TP (g/L) 24.20ab 26.40ab 21.50b 25.35ab 27.25a 25.35ab ±0.73

GLOB (g/L) 10.80b 13.95a 12.70ab 13.55ab 14.30a 15.20a ±0.45

ALB 13.40a 12.45ab 8.80b 11.80ab 12.95a 10.15ab ±0.64

AST (U/L) 45.00ab 40.00ab 50.50a 46.50ab 46.00ab 28.50b ±2.56

ALT (U/L) 28.00 8.00 16.50 26.00 22.00 12.00 ±2.76

ALP (U/L) 394.00 360.00 376.00 376.50 381.50 330.00 ±15.16

CHOLES(mmol/L) 3.05b 4.05a 3.80a 4.40a 4.00a 4.20a ±0.13

TG (Mmol/L) 0.30b 0.62ab 0.83a 0.85a 0.58ab 0.84a ±0.07

CREAT (µmol/L) 29.20a 20.10c 25.10b 15.10d 21.10c 29.05a ±1.28

ACP (U/L) 4.85 4.20 5.80 2.95 4.75 4.35 ±0.45 a, b, c, d = means within the same row bearing different superscripts differ significantly. TP: Total Protein. GLOB: Globulin. ALB:

Albumin. AST: Aspartate Transaminase. ALT: Alanine Transaminase. CHOLES: Cholesterol. TG: Triglyceride. REAT: Creatinine. ACP:

Acid Phosphatase

International Journal of Applied Research and Technology 38

Table 7: Effect of treated jkm diet on lactobacillus and escherichia coli population in the git of broiler chicks.

Treatment Lactobacillus (103) E.coli (103)

D1 30.00 1.6

D2 3.0 ND

D3 1.5 ND

D4 0.55 ND

D5 0.35 ND

D6 5.0 ND

ND: not detected