Journal of Applied Phycology

https://doi.org/10.1007/s10811-018-1498-y

Production performance, immune response and carcass traits of broiler

chickens fed diet incorporated with Kappaphycus alvarezii
S. S. N. Qadri 1 & A. Biswas 1 & A. B. Mandal 1 & M. Kumawat 1 & R. Saxena 1 & A. M. Nasir 1

Received: 2 February 2018 / Revised and accepted: 23 April 2018

# Springer Science+Business Media B.V., part of Springer Nature 2018

Abstract
This study evaluates the production performance, immune responses and carcass traits in broiler chickens (0 to 42 days) fed diet
with incorporated Kappaphycus alvarezii (AF-KWP). Day-old chicks (280) were randomly distributed into 35 groups with 8
chicks in each group (7 dietary treatments × 5 replicates). Seven experimental diets were formulated by adding 0 (control—T1),
0.25 AF-KWP (T2), 0.50 AF-KWP (T3), 0.75 AF-KWP (T4), 1.00 AF-KWP (T5), 1.25 AF-KWP (T6) and 1.50 AF-KWP (T7)
respectively. Body weight gain (g) significantly (P < 0.05) improved in the growing phase (21–42 days) and overall value (0–
42 days); however, feed intake (g) differed significantly (P < 0.05) in T7 and T6 groups only in the growing (21–42 days) phase,
compared to control and other treated groups. Feed conversion ratio (FCR) did not differ significantly (P > 0.05) by the feeding of
diets containing different levels of AF-KWP. Haemagglutination (HA) titre and cell-mediated immunity (CMI) were significantly
increased (P < 0.05) in AF-KWP supplemented groups as compared to the control group. No significant (P > 0.05) difference
was recorded in the percentage of carcass traits and organ weights but significant improvement (P < 0.05) was recorded in cut up
parts (percentage of live wt.) between the control and some AF-KWP-supplemented groups. From the results, it may be
concluded that K. alvarezii (AF-KWP) can be incorporated at 1.25% level in broiler diets for improved performance,
immuno-responsiveness and breast yield in broiler chickens.

Keywords Kappaphycus . Rhodophyta . Production performance . Immunity . Carcass trait . Chicken

Introduction algae or weeds which contribute to inhibition or suppression

of free radical generation (Athukorala et al. 2006; Wells et al.
Algae are a large and diverse group of plant-like organisms 2017). Kappaphycus alvarezii is rich in enzymes, nutrients,
ranging from unicellular to multicellular forms. Kappaphycus minerals, calcium, iron, multitude fibres and jelly-forming
alvarezii (AF-KWP) is a red alga which is consumed by coast- polysaccharides. Kappaphycus alvarezii contains 64.2% car-
al people in the Tamil Nadu, India, and elsewhere (Mantri bohydrates, 4.5% protein, 0.9% fat, 1.07% calcium,
et al. 2017; Peryasami and Subba Rao 2017). Fresh and dried 9.3 mg kg−1 iron, 1520 mg kg−1 magnesium and 22 mg kg−1
seaweeds are utilised as human food. Soluble polysaccharides niacin as well as 0.180 mg g−1 fresh weight chlorophyll
have hyper-cholesterolaemic and hypoglycaemic effects, (Abirami and Kowsalya 2011). Carotenoids in seaweeds
whereas water insoluble polysaccharides decrease digestive have beneficial effects by acting either as an antioxidant
tract transit time (Santoso et al. 2004) and thus retention time or pro-oxidant depending on the environment and
is increased, which may be beneficial for improving digest- K. alvarezii has been shown to have a total carotenoid
ibility of nutrients. Seaweeds have recently received signifi- content of 0.634 mg g −1 fresh weight (Abirami and
cant attention for their potential as natural antioxidants attrib- Kowsalya 2011). Kappaphycus alvarezii has been intro-
uted to carotenoids, tocopherols and polyphenols present in duced throughout the warm tropics for commercial culti-
vation (Hurtado et al. 2014). It is a major producer of
kappa-carrageenan, which is used for medicinal purposes
* A. Biswas
and as a homogeniser in milk products, toothpaste and
[email protected] jelly. Thus, the selected seaweed biomass could well be
included in practical poultry feed to combat deficiency
1 micronutrients, as an antioxidant and to produce designer
Avian Nutrition and Feed Technology Division, ICAR-Central Avian
Research Institute, Izatnagar, U.P. 243122, India poultry products with functional attributes.
 J Appl Phycol

The present research work studied the growth performance, was kept at 30 °C for the first week and then decreased 2 °C
immune response and meat qualities as influenced by per week.
Kappaphycus alvarezii in the diet of broiler chickens.

Determined the nutrient content

Material and methods The nutrient contents of AF-KWP are as follows: moisture
05.18%; crude protein 10.26%; fat 04.68%; crude fibre
Experimental birds 02.91; total ash 60.24%. Moisture content of the sample was
estimated by heating it in an oven to a constant weight at 100–
Two hundred and eighty (280) straight-run day-old broiler 105 °C under atmospheric pressure. Ash is the inorganic or
chicks were housed and randomly distributed into 35 groups, mineral component of the sample left after complete ignition
each of 8 chicks (7 treatments × 5 replicates). The experiment of the sample at 600 °C in a muffle furnace. The crude protein
had a complete randomised design (Snedecor and Cochran (%) in AF-KWP was determined by a modified macro-
1994). The experiment was conducted strictly in accordance Kjeldahl method (Jackson 1973), while phosphorus content
with the guidelines of the Institutional Animal Ethics was determined by a vanado-molybdophosphoric acid yellow
Committee (IAEC). The temperature of the experimental shed colour method in a HNO 3 system with a colourimeter

Table 1 Ingredient and nutrient

composition of pre-starter diet (0– Ingredients (kg/100 kg)−1 Pre-starter diet (0–14 days)
14 days of age)
T1 T2 T3 T4 T5 T6 T7

Maize, yellow 48.915 48.665 48.415 48.165 47.915 47.615 47.365

AF-KWP 0 0.25 0.5 0.75 1 1.25 1.5
Soya bean meal (S. ext.) 43.1 43.1 43.1 43.1 43.1 43.1 43.1
Rapeseed meal (S. ext.) 3 3 3 3 3 3 3
Oil 1.6 1.6 1.6 1.6 1.6 1.65 1.65
Limestone powder 0.9 0.9 0.9 0.9 0.9 0.9 0.9
Di-calcium phosphate 1.7 1.7 1.7 1.7 1.7 1.7 1.7
Salt 0.3 0.3 0.3 0.3 0.3 0.3 0.3
DL-meth 0.17 0.17 0.17 0.17 0.17 0.17 0.17
L-lysine HCl 0.01 0 0 0 0 0 0
TM. premix1* 0.1 0.1 0.1 0.1 0.1 0.1 0.1
Vit. premix2** 0.15 0.15 0.15 0.15 0.15 0.15 0.15
B complex premix 0.015 0.015 0.015 0.015 0.015 0.015 0.015
Ch. chloride (60%) 0.05 0.05 0.05 0.05 0.05 0.05 0.05
Calculated value
Crude protein (%)*** 22.50 22.51 22.51 22.51 22.51 22.51 22.52
M energy (kcal/kg−1)## 2899.89 2899.52 2899.14 2898.77 2898.39 2900.64 2900.27
Calcium (%)# 1.02 1.02 1.02 1.02 1.02 1.01 1.01
Available P (%)# 0.45 0.45 0.45 0.45 0.45 0.45 0.45
Lysine (%)## 1.28 1.28 1.28 1.28 1.28 1.28 1.28
Methionine (%)## 0.56 0.56 0.56 0.56 0.56 0.56 0.56
Threonine (%)## 1.00 1.00 1.00 1.00 1.00 1.00 0.99

AF-KWP Kappaphycus alvarezii and red sea weed based formulation

*Trace mineral premix supplied Mg 300, Mn 55, I 0.4, Fe 56, Zn 30 and Cu 4 mg kg−1 diet
**Vitamin premix supplied vitamin A, 8250 IU; vitamin D3, 1200 ICU; vitamin K, 1 mg; vitamin E, 40 IU;
vitamin B1, 2 mg; vitamin B2, 4 mg; vitamin B12, 10 mcg; niacin, 60 mg; pantothenic acid, 10 mg; choline,
500 mg kg−1 diet
***Analysed values
#
Calculated based on analysed ingredient values
##
Calculated tabulated values
J Appl Phycol

(Jackson 1973). Fat content was evaluated by a wet digestion ad lib feeding, but with care to avoid spoilage and wastage of
method with Soxtec apparatus (Jackson 1973). feed. Clean and fresh water was always made available in
suitable troughs to all the birds during the study period.
Formulation of experimental diets

Seven experimental diets were formulated by adding 0 (con- Production performance

trol—T1), 0.25 AF-KWP (T2), 0.50 AF-KWP (T3), 0.75 AF-
KWP (T4), 1.00 AF-KWP (T5), 1.25 AF-KWP (T6) and 1.50 Body weight changes were recorded periodically (0–3 and 4–
AF-KWP (T7) and all the diets were isocaloric and iso-nitrog- 6 weeks) during the experimental period to ascertain the
enous. The ingredient and chemical compositions of the diets weekly and overall body weight gain. A weighed quantity of
used in this study are presented in Tables 1, 2 and 3. the respective diet was offered ad lib daily to quadruplicate
groups of each dietary regimen in the morning and the residue
Feeding and watering was weighed next day on a daily basis in order to arrive at
overall feed intake. Based on the data pertaining to the feed
Weighed amounts of each test diet used during the starting intake (FI) and body weight gain (BWG), the weekly and
period (0–3 weeks) and finishing period (4–6 weeks) were period-wise feed conversion ratio (FCR) of birds was
offered daily in five replicates of eight chicks each to ensure determined.

Table 2 Ingredient and nutrient

composition of starter diet (14– Ingredients (kg (100 kg)−1) Starter diet (14–21 days)
21 days of age)
T1 T2 T3 T4 T5 T6 T7

Maize, yellow 51.485 51.235 50.985 50.735 50.485 50.235 49.985

AF-KWP$ 0 0.25 0.5 0.75 1 1.25 1.5
Soya bean meal (S. ext.) 40.3 40.3 40.3 40.3 40.3 40.3 40.3
Rapeseed meal (S. ext.) 3 3 3 3 3 3 3
Oil 2 2 2 2 2 2 2
Limestone powder 0.9 0.9 0.9 0.9 0.9 0.9 0.9
Di-calcium phosphate 1.6 1.6 1.6 1.6 1.6 1.6 1.6
Salt 0.3 0.3 0.3 0.3 0.3 0.3 0.3
DL-meth 0.1 0.1 0.1 0.1 0.1 0.1 0.1
L-lysine HCl 0.1 0.1 0.1 0.1 0.1 0.1 0.1
TM. premix1* 0.15 0.15 0.15 0.15 0.15 0.15 0.15
Vit. premix2** 0.015 0.015 0.015 0.015 0.015 0.015 0.015
B complex premix 0.05 0.05 0.05 0.05 0.05 0.05 0.05
Ch. chloride (60%) 51.485 51.235 50.985 50.735 50.485 50.235 49.985
Calculated value
Crude protein (%)*** 21.51 21.52 21.52 21.52 21.52 21.53 21.53
M energy (kcal kg−1)## 2952.07 2951.69 2951.32 2950.94 2950.57 2950.19 2949.82
Calcium (%)# 0.98 0.98 0.98 0.98 0.98 0.98 0.98
Available P (%)# 0.43 0.43 0.43 0.43 0.43 0.43 0.43
Lysine (%)## 1.21 1.21 1.21 1.21 1.21 1.21 1.21
Methionine (%)## 0.48 0.48 0.48 0.48 0.48 0.48 0.48
Threonine (%)## 0.96 0.96 0.95 0.95 0.95 0.95 0.95

AF-KWP Kappaphycus alvarezii and red sea weed based formulation

*Trace mineral premix supplied Mg 300, Mn 55, I 0.4, Fe 56, Zn 30 and Cu 4 mg kg−1 diet
**Vitamin premix supplied vitamin A, 8250 IU; vitamin D3, 1200 ICU; vitamin K, 1 mg; vitamin E, 40 IU;
vitamin B1, 2 mg; vitamin B2, 4 mg; vitamin B12, 10 mcg; niacin, 60 mg; pantothenic acid, 10 mg; choline,
500 mg kg−1 diet
***Analysed values
#
Calculated based on analysed ingredient values
##
Calculated tabulated values
 J Appl Phycol

Table 3 Ingredient and nutrient

composition of finisher diet (21– Ingredients (kg (100 kg)−1) Finisher diet (21–42 days)
42 days of age)
T1 T2 T3 T4 T5 T6 T7

Maize, yellow 57.085 56.835 56.585 56.335 56.085 55.835 55.585

AF-KWP 0 0.25 0.5 0.75 1 1.25 1.5
Soya bean meal (S. ext.) 34.2 34.2 34.2 34.2 34.2 34.2 34.2
Rapeseed meal (S. ext.) 3 3 3 3 3 3 3
Oil 2.7 2.7 2.7 2.7 2.7 2.7 2.7
Limestone powder 0.8 0.8 0.8 0.8 0.8 0.8 0.8
Di-calcium phosphate 1.5 1.5 1.5 1.5 1.5 1.5 1.5
Salt 0.3 0.3 0.3 0.3 0.3 0.3 0.3
DL-meth 0.1 0.1 0.1 0.1 0.1 0.1 0.1
L-lysine HCl 0.1 0.1 0.1 0.1 0.1 0.1 0.1
TM. premix1* 0.15 0.15 0.15 0.15 0.15 0.15 0.15
Vit. premix2** 0.015 0.015 0.015 0.015 0.015 0.015 0.015
B complex premix 0.05 0.05 0.05 0.05 0.05 0.05 0.05
Ch. chloride, 60% 57.085 56.835 56.585 56.335 56.085 55.835 55.585
Calculated value
Crude protein (%)*** 19.50 19.50 19.50 19.51 19.51 19.51 19.52
M energy (kcal kg−1)## 3051.03 3050.65 3050.28 3049.90 3049.53 3049.15 3048.78
Calcium (%)# 0.90 0.90 0.90 0.90 0.90 0.90 0.90
Available P (%)# 0.40 0.40 0.40 0.40 0.40 0.40 0.40
Lysine (%)## 1.06 1.06 1.06 1.06 1.06 1.06 1.06
Methionine (%)## 0.45 0.45 0.45 0.45 0.45 0.45 0.45
Threonine (%)## 0.86 0.86 0.86 0.86 0.86 0.86 0.86

AF-KWP Kappaphycus alvarezii and red sea weed based formulation

*Trace mineral premix supplied Mg 300, Mn 55, I 0.4, Fe 56, Zn 30 and Cu 4 mg kg−1 diet
**Vitamin premix supplied vitamin A, 8250 IU; vitamin D3, 1200 ICU; vitamin K, 1 mg; vitamin E, 40 IU;
vitamin B1, 2 mg; vitamin B2, 4 mg; vitamin B12, 10 mcg; niacin, 60 mg; pantothenic acid, 10 mg; choline,
500 mg kg−1 diet
***Analysed values
#
Calculated based on analysed ingredient values
##
Calculated tabulated values

Immunological studies The foot web index (FWI) was used as an index of the cell-
mediated immune response. During the fifth week (35 days),
To investigate the effect on the humoral immune response, three birds from each replicate of the treatment groups were
three birds were selected from each of the replicated selected and 0.2 mL phyto-haemagglutinin (PHAP) mitogen
groups (15 birds per dietary treatment providing 105 birds (1 mg mL−1 PBS) was injected intra-dermally into the left foot
in all) at 28 days of age and were inoculated intravenously web. Sterile PBS (0.2 mL) was injected into the right foot web
with 1.0 mL of 1% suspension of sheep red blood cell to serve as a control. A micrometer was used to measure
(SRBC). Blood samples were obtained from the jugular changes in the thickness of both foot webs. Measurements
vein from all SRBC injected birds at 0 and 6 days post- were made at 0 and 24 h after the injection, as described by
inoculation. All samples were incubated at 37 °C for 1 h to Cheng and Lamont (1988). Foot web swelling was calculated
aid clotting and retraction, then centrifuged at 15000×g for by subtracting skin thickness at 24 h post-injection from that at
5 min for collection of sera. All the microtitre plates (U- 0 h pre-injection.
bottomed) were rinsed with phosphate-buffered saline
(PBS; pH 7.6) then dried before the haemagglutination Carcass traits and cut up part weight
antibody (HA) titre was estimated by a micro-
haemagglutination method (Siegel and Gross 1980) using At the end of the experiment, three birds from each replicate of
twofold serial dilutions of sera. the treatment (15 birds per dietary treatment, n = 105) were
J Appl Phycol

Table 4 Effects of dietary inclusion of Kappaphycus alvarezii and red sea weed based formulations on production performance of broiler chickens
(N = 15)

Group Dietary treatments Body weight gain (g) Feed intake (g) Feed conversion ratio

0–21 days 21–42 days 0–42 days 0–21 days 21–42 days 0–42 days 0–21 days 21–42 days 0–42 days

T1 Control diet 492.7 915.6a 1408.3a 845.6 1884.1a 2575.8 1.71 2.05 1.83
a a
T2 T1 + 0.25% AF-KWP 495.8 937.8 1433.6 862.8 1896.1a 2609.1 1.74 2.02 1.82
T3 T1 + 0.50% AF-KWP 505.5 936.2a 1442.2a 879.6 1900.4a 2509.5 1.74 2.03 1.74
T4 T1 + 0.75% AF-KWP 490.9 960.1ab 1450.9a 829.6 1955.4ab 2539.0 1.69 2.04 1.75
T5 T1 + 1.00% AF-KWP 484.9 964.7ab 1449.6a 806.4 1978.6ab 2568.9 1.66 2.05 1.77
T6 T1 + 1.25% AF-KWP 533.9 1026.2b 1560.1b 865.0 2124.5b 2699.0 1.62 2.07 1.73
T7 T1 + 1.50% AF-KWP 513.9 1021.1b 1535.1b 827.5 2052.5b 2640.4 1.61 2.01 1.72
SEM – 6.20 11.5 19.56 5.9 23.12 28.45 0.001 0.002 0.001
P value – 0.319 0.038 0.027 0.057 0.033 0.240 0.052 0.062 0.051

Mean values bearing the same superscript in a column did not differ significantly (P < 0.05)

selected randomly and killed to determine the carcass traits, Immune response
cut up parts and relative weight of the lymphoid organs (bursa
of Fabricius and thymus) and the liver. The thymus tissue was The results of immune response (cell mediated and humoral)
carefully dissected from each side of the neck to ensure com- assessed through PHAP and HA methods are presented in
plete removal. Organ relative weights were measured to the Table 5. Cell-mediated immune response (CMI) measured
nearest 0.01 g. by foot web index thickness differed significantly (P < 0.05)
among control and dietary treatment groups. The T2, T3 and
Statistical analysis T4 treatment groups showed intermediate values and were
statistically similar to the control. Humoral immune response
Data were analysed using statistical software SPSS-20 version expressed as HA titre (log2) also differed significantly
and followed standard (Snedecor and Cochran 1994) proce- (P < 0.05) between control and other dietary treatment groups.
dures for ANOVA. Duncan’s (Duncan 1955) multiple range The highest value was found in T7 (1.50% AF-KWP) follow-
test by comparing means for significant (P < 0.05) differences ed by statically similar T6 (1.25% AF-KWP) and lowest value
was used. in T1 (control) followed by T2, whereas T3, T4 and T5 showed
intermediate values and did not differ significantly (P > 0.05)
from each other. Both cell-mediated and humoral immune
responses showed an increasing trend from T1 to T7 group.
Results
Table 5 Effects of dietary inclusion of Kappaphycus alvarezii and red
Growth performance sea weed based formulations on immune response of broiler chickens
(N = 15)

The results pertaining to body weight gain, feed intake and Group Dietary treatments Immune response
feed conversion ratio (FCR) are presented in Table 4. Body
weight gain did not differ significantly (P > 0.05) due to die- CMI (mm) HA titre (log2)
tary treatment groups during 0–21 days of age. However, the
T1 Control diet 0.50a 1.92a
body weight gain differed significantly (P < 0.05) during the
T2 T1 + 0.25% AF-KWP 0.51a 1.94a
finishing phase (21–42 days) and overall (0–42 days) with
T3 T1 + 0.50% AF-KWP 0.52ab 1.98ab
highest body weight gain in T6 (1.25% AF-KWP) followed
T4 T1 + 0.75% AF-KWP 0.52ab 2.02ab
by T7 (1.50% AF-KWP) and lowest body weight gain in T1
T5 T1 + 1.00% AF-KWP 0.53b 2.03ab
(control). The feed intake among the treatments did not differ
T6 T1 + 1.25% AF-KWP 0.53b 2.05b
significantly (P > 0.05) in the starting growth phase; however,
T7 T1 + 1.50% AF-KWP 0.54b 2.06b
during the finishing period (21–42 days), significant differ-
SEM – 0.05 0.07
ences were seen among the treatment groups. The feed con-
P value 0.013 0.038
version ratio (FCR) did not differ significantly (P > 0.05);
however, the FCR improved linearly with the increased level Mean values bearing the same superscript in a column did not differ
of AF-KWP with the highest value in T1 and lowest in T7. significantly (P < 0.05)
 J Appl Phycol

Table 6 Effects of dietary inclusion of Kappaphycus alvarezii and red sea weed based formulations on carcass traits and organ weight of broiler
chickens (N = 15)

Group Dietary treatments Live weight (g) Dressing yield (%) Eviscerated yield (%) Heart (%) Liver (%) Gizzard (%)

T1 Control diet 1589.17 68.76 63.76 0.61 2.26 2.13

T2 T1 + 0.25% AF-KWP 1633.33 69.54 64.44 0.65 2.28 2.17
T3 T1 + 0.50% AF-KWP 1664.17 69.99 64.74 0.66 2.39 2.10
T4 T1 + 0.75% AF-KWP 1674.00 70.17 64.88 0.66 2.40 2.20
T5 T1 + 1.00% AF-KWP 1676.92 70.25 64.81 0.67 2.44 2.22
T6 T1 + 1.25% AF-KWP 1686.33 70.62 65.06 0.67 2.51 2.33
T7 T1 + 1.50% AF-KWP 1713.33 72.68 67.38 0.69 2.54 2.36
SEM – 16.86 0.34 0.41 0.02 0.04 0.05
P value – 0.525 0.079 0.124 0.055 0.065 0.072

Mean values bearing the same superscript in a column did not differ significantly (P < 0.05)

Carcass traits and cut up parts microalgae has a positive effect on body weight gain in broiler
chickens. However, the results of the present study were not in
The results pertaining to the effects of feeding of test diets agreement with findings of Evans et al. (2015) who observed
(AF-KWP) on carcass traits (%), organ weight (%) and cut no significant differences among the dietary microalgae-
up parts (% of live weight) are shown in the Tables 6 and 7. No treated groups. There are very few reports available in the
significant difference (P > 0.05) was observed in carcass char- literature regarding the effect of AF-KWP in poultry diets.
acteristics in terms of dressing and eviscerated percentage and In the present study, AF-KWP had a positive effect on growth
different organ weights among the control and dietary treat- performance, which was associated with the manipulation of
ment groups. However, higher dressing and eviscerated per- improved immunity. Data for feed intake and feed conversion
centage were recorded in T7 group flowed by T6 and T5 group. ratio (FCR) in the present study differ from the findings of
On the other hand, cut up parts (percentage of live wt.) in Ross and Dominy (1990) who found no significant difference
terms of thigh, breast, back and wings differed significantly in feed intake and FCR of broilers. However, Mariey et al.
(P < 0.05). Breast yield was higher at high levels of AF-KWP. (2014) and Evans et al. (2015) reported that microalgae had a
positive effect on feed intake and FCR which corroborated the
present findings. The specific reports regarding feed intake
and FCR were not available concerning supplementation of
Discussion K. alvarezzi to a basal diet.

Growth performance
Immune response
The results of the present study are in agreement with the
findings of Mariey et al. (2014), Shanmugapriya et al. In the present study, humoral and cell-mediated response
(2015) and Byoung et al. (2016), who reported that 1% were significantly improved in T7 (1.5%) followed by T6

Table 7 Effects of dietary

inclusion of Kappaphycus Group Dietary treatments Thigh Breast Back Wings Neck Drumstick
alvarezii and red sea weed based
formulations on cut up part yield T1 Control diet 9.57a 16.04a 19.86b 8.44a 4.46 10.33
(% of live weight) of broiler T2 T1 + 0.25% AF-KWP 9.67a 16.25a 19.20ab 8.79ab 4.20 10.45
chickens (N = 15) T3 T1 + 0.50% AF-KWP 9.79a 16.55a 18.37ab 8.92ab 4.48 10.34
T4 T1 + 0.75% AF-KWP 9.83a 16.80ab 19.86b 9.05ab 4.20 10.10
T5 T1 + 1.00% AF-KWP 10.12ab 16.92ab 19.20ab 9.07ab 4.32 10.21
T6 T1 + 1.25% AF-KWP 10.25b 17.25b 19.20ab 9.87b 4.48 10.60
T7 T1 + 1.50% AF-KWP 10.30b 18.35b 18.70ab 9.84b 4.67 10.84
SEM – 0.37 0.56 0.42 0.22 0.12 0.67
P value – 0.042 0.049 0.024 0.020 0.089 0.120

Mean values bearing the same superscript in a column did not differ significantly (P < 0.05)
J Appl Phycol

(1.25%). Kang et al. (2013) also reported that the Compliance with ethical standards
microalga Chlorella has some beneficial effect on immune
characteristics, i.e. the number of WBCs, lymphocytes and All institutional and national guidelines for the care and use of laboratory
animals were followed.
IgA, IgM and IgG concentrations. Likewise, Kotrbacek
et al. (1994) found that broilers fed a diet of 0.5%
Conflict of interest The authors declare that they have no conflict of
microalgae significantly (P < 0.05) increased the phagocyt- interest.
ic activity of leucocytes and lymphatic tissue development.
However, the present results were not in accordance with
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