present study, vitamin C is known to be able. Vitamin C is able to donate electrons and hydrogen to the free radicals;
consequently free radicals become relatively stable. Criteria for blood parameter in present study included ratio of
heterophile-lymphocytes (H/L) and blood plasma protein. H/L ratio is the main stress indicator in poultry, because the
higher the ratio number, the severe stress level is experienced, and vice versa (Kusnadi, 2009). Plasma protein is a
secondary indicator of blood parameter besides the H/L ratio, because it is an indication of an adequacy supply of amino
acids for antibody formation. The general phenomenon is that the higher plasma protein, the better blood parameter of
chickens that support productivity.
MATERIALS AND METHODS
Experimental animal and diet
The experiment used 12-week old of 64 male birds obtained from in situ maintenance (native habitat) in Cianjur,
West Java, divided into 4 groups based on body weight (group 1: 740–910 g, group 2: 910–1,080 g, group 3: 1,080–
1,250 g, and group 4: 1,250–1,420 g) and reared for 6 weeks. Basal diet composed of rice bran, yellow corn, white bran,
soybean meal, meat bone meal, and mix mineral which are prepared with a crude protein content of 14.57% and
metabolizable energy (ME) of 2,892.03 kcal/kg (Table 1). Lime juice according to treatment level at 1, 2 and 3% were
mixed with approximately 20 g of diet and given in the morning, to ensure all lime juice can be complete consumed, and
followed by diet without lime juice to fulfill daily requirement. Experimental diets were provided from 13 week to 18 week
old after condition period for one week (12-13 week).
Table 1 - Compositions and nutrient content for formulated diet.
Ingredients
Rice bran
Yellow corn
Wheat bran
Soybean meal
Meat bone meal
Mineral and vitamin mix3
Total
Metabolizable energy (kcal/kg)2
Crude Protein1
Crude Fiber1
Compositions (%)
20.00
55.00
10.00
6.00
4.00
5.00
100.00
2,892.03
14.57
7.37
Ether extract1
Calcium1
6.57
0.81
Total phosphor
0.55
1
2
Sources: Based on chemical analysis at the Laboratory of Feed and Nutrition Science, Diponegoro University; Based on Bolton (1967) as
follow: 40.81 {0.87 [crude protein + 2.25 crude fat + nitrogen free extract] + 2.5}; Provided minerals per kg of the feed: calcium 32.5%,
3
phosphor 1%, iron 6 g, mangan 4 g, iodine 0,075 g, copper 0,3, zinc 3,75 g, vitamin B12 0,5 mg, vitamin D3 50,000 IU.
Experimental Parameters
Observed parameters were heterophile, lymphocytes, heterophile-lymphocytes ratio, plasma protein and lymphoid
organs (spleen and bursa of fabricius) measured at fifth the last week of experimental rearing period. Blood samples were
taken through the brachial vein with 3 mL syringe. Blood was inserted into tube containing anticoagulant as was used
sample for heterophile-lymphocyte test and total plasma protein based on Enzyme-Linked Immunosorbent Assay (ELISA)
method. Chickens were further decapitated and dissected to obtain lymphoid organs (spleen and bursa of fabricius) to
measure relative weight by dividing with final body weight and multiplied by 100%.
Experimental design and statistical analysis
The study was arranged in a randomized block design (RBD) based on body weight and as follows:
K1 = body weight range 740 – 910 g with average 835.51 ± 49.48 g
K2 = body weight range > 910 – 1,080 g with average 985.69 ± 42.04 g
K3 = body weight range >1,080 – 1,250 g with average 1,160.63 ± 55.69 g
K4 = body weight range >1,250 – 1,420 g with average 1,369.06 ± 65.97 g
The study was designed in 4 treatments and 4 groups as replications (4 birds each). The treatment applied:
T0 = formulated diet
T1 = formulated diet + lime juice 1%
T2 = formulated diet + lime juice 2%
T3 = formulated diet + lime juice 3%
Data subjected to analysis of variance to determine the effect of treatment. If the treatment indicated significant
104
Citation: Krismiyanto L, Wahyuni HI and Suthama dN (2021). Effect of lime (Citrus aurantifolia) juice as source of vitamin c on performance and some blood parameters