Indian Journal of Animal Sciences 86 (11): 1318–1322, November 2016/Article

Interaction of oxidative feed sanitizer with dietary methionine on growth and

immune-responsiveness of broiler chickens
RAKESH THAKUR1, A B MANDAL2 and A S YADAV3

Central Avian Research Institute, Izatnagar, Uttar Pradesh 243 122 India

Received: 25 February 2016; Accepted: 27 July 2016

ABSTRACT
Acidified sodium chlorite (ASC), a well known sanitizer, reduces microbial load and improves energy
bioavailability from fiber/lignin rich feedstuffs. However, ASC also reacts with sulphur containing amino acid
methionine, the first limiting amino acid in maize-soy based diet. So the present experiment was conducted to
assess the interaction of ASC with dietary methionine in terms of growth performance, meat quality and immune-
competence of broiler chicks. A basal diet was formulated without methionine supplementation (otherwise adequate
in all nutrients) and then treated with 3 levels of ASC (0, 100 and 250 ppm). Each ASC treated basal diet was
supplemented without or with DL-methionine to meet 100 and 120% of NRC requirement of methionine in a 3 × 3
factorial design. Day-old broiler chicks (288) were divided into 36 groups of 8 chicks each and each dietary treatment
was offered to 4 replicated groups. At 6 weeks of age, methionine supplementation produced significant increase in
live body weight, weight gain and feed conversion ratio. Among meat quality parameters, thiobarbituric acid reactive
substance value increased significantly upon ASC treatment and reduced by methionine supplementation at 120%.
Among cut-up parts, neck yield increased significantly with increased level of methionine. Increase in methionine
level also produced improvement in humoral and cell mediated immunity. Thus, it may be concluded that ASC
treatment deteriorated oxidative stability of meat while methionine supplementation improved broiler performance,
meat quality and immunocompetence.

Key words: Acidified sodium chlorite, Broilers performance, Immune-competence, Meat quality, Methionine

Acidified sodium chlorite (ASC) produced by adding requirement was higher for maximum feed efficiency and
citric acid to an aqueous solution of sodium chlorite, was breast meat yield than for obtaining maximum weight gain.
approved by FDA as an antimicrobial intervention treatment Chlorine dioxide, which has oxidizing properties similar
for poultry carcasses, poultry carcass parts, red meat to ASC, also reported to react with amino acids, especially
carcasses, red meat parts and organs, seafood, and raw those containing sulphur in their structure (Tan et al. 1987).
agricultural commodities owing to its strong biocidal Thakur and Mandal (2015) recorded a small statistically
properties. ASC also reduces microbial load in various feed non-significant reduction in methionine digestibility from
ingredients and improves the energy bioavailability from feedstuffs like soybean meal, meat-cum-bone meal and
fiber/lignin rich feedstuff and maize-soy mixture (Thakur sunflower meal indicating ASC had a tendency to interact
et al. 2014) in chicken. But being oxidative in nature, ASC with methionine and in turn rendering it less bioavailable.
may interact with chemical groups in amino acids, fatty So the present experiment was conducted to assess the
acids, which are susceptible to oxidation thus, may interaction of acidified sodium chlorite with dietary
deteriorate quality of feed and ultimately may induce methionine for growth performance, meat quality and
oxidative stress to the consumers. Methionine is the first immune-competence of broiler chicks.
limiting amino acid in commercial maize-soy based mixed
MATERIALS AND METHODS
feeds, and need to be supplemented to improve dietary
amino acid balance and promotes greater protein build up Acidified sodium chlorite was prepared by adding citric
in poultry (Bunchasak 2009). Schutte and Pack (1995) acid in powder form, to an aqueous solution (625 g/l) of
reported that the total sulphur amino acid (TSAA) sodium chlorite (80%) to attain a pH of 3.5. This stock
solution had a concentration of 500 ppm/ml and
Present address: 1Subject Matter Specialist -Animal Science,
immediately before use, the stock solution of ASC was
([email protected]), Krishi Vigyan Kendra Mandi at
Sundernagar, Himachal Pradesh. 2Principal Scientist and Head diluted in water and thoroughly mixed with feed to achieve
([email protected]), Avian Nutrition and Feed Technology. different concentrations.
3 Principal Scientist ([email protected]), Post-Harvest To assess interaction of ASC with dietary methionine,
Technology. dietary treatments were formulated using a basal diet

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November 2016] ACIDIFIED SODIUM CHLORITE AND DIETARY METHIONINE IN BROILERS 1319

(otherwise adequate in all nutrients except methionine) requirement had no significant improvement on broiler
treated with 3 levels of ASC (0, 100 and 250 ppm). Each performance, but decreased abdominal fat significantly.
diet was supplemented without (methionine deficient diet) Hickling et al. (1990) fed broilers 2 methionine levels (100
or with DL-methionine to meet 100 and 120% of methionine or 116% of NRC, 1984) and found that increasing
requirement as suggested by NRC (1994) in a 3 × 3 factorial methionine significantly improved body weight at 6 week
design. Day-old broiler chicks (288) were divided into 36 of age but not at 3 weeks of age. This variation might be
groups of 8 chicks each. Each dietary treatment was offered due to difference in growth potential of the germplasm
to 4 replicated groups of chicks during their starting (0–3 (synthetic male line) used in this study.
week) and finishing (4–6 week of age) growth phases. Feed intake was not affected by either ASC treatment or
Body weight changes and feed intake were recorded methionine supplementation. However, in feed conversion
weekly, while mortality of birds as and when occurred. ratio (FCR) methionine supplementation resulted in
Humoral immunity and cell mediated immunity were significant improvement during 5–6 (P<0.001) and 0–6
determined as per Vander Zijp (1983) and Corrier and week’s (P<0.003) growth phases (Table 2). This
Deloach (1990), respectively. At the end of 42 days of age, improvement was due to better growth and weight gain, as
6 birds from each treatment were sacrificed. The bursa, the feed intake was almost similar amongst the treatment
spleen and upper 3 lobes of the thymus from the left side of groups. Hassan et al. (2003) showed that addition of
the neck were dissected out, weighed and expressed as methionine (0.15%) in broiler diets improved feed
relative (mg/100 g) to live weight. The following carcass conversion of the birds compared to the basal. Schutte and
parameters, viz. deblooded weight, defeathered weight and Pack (1995) after feeding a range of methionine and total
cut-up part yields (breast, thigh, drumsticks, back, neck and sulphur amino acid (TSAA) levels for 14–38 days, estimated
wing) were recorded and expressed as percent of live a TSAA requirement of 0.84% for weight gain and 0.88%
weight. The meat quality in terms of pH, water holding for FCR, this partially justifies the small improvement in
capacity (WHC), and oxidative status (in terms of FCR in diet with methionine 120% level over 100%. ASC
thiobarbituric acid reactive substance level, TBARS) was treatment did not affect FCR over any of the growth phase.
studied in breast muscles. TBARS, expressed as Carcass characters and meat quality; Carcass traits and
malonaldehyde is a good index reflecting the degree of cut-up parts yield expressed as percentage of dressed weight
oxidation (Lohakare et al. 2005). The responses of ASC
and methionine were analyzed statistically as per Snedecor Table 1. The ingredient (%) and nutrient composition of broiler
and Cochran (1989). basal starter and finisher ration

RESULTS AND DISCUSSION Ingredient Starter ration Finisher ration

(0-4 wk) (4-6 wk)
Growth performance and feed conversion ratio: The live
weight and weight gain at fourth week of age was Maize 56 57.7
statistically similar amongst all the treatments. However at Sunflower 6 6
sixth week of age, methionine levels 100 and 120% De-oiled rice bran 4.73 11
produced significant increase in live body weight (P<0.032) Soybean meal 30 22.4
and weight gain (P<0.036) compared to basal methionine Limestone 1.2 1.5
level (Table 2). Dicalcium phosphate 1.4 0.8
Methionine is the first limiting amino acid in maize-soy Trace minerals* 0.1 0.1
based poultry diets and methionine supplementation is Vitamin premix** 0.1 0.1
required to meet birds’ requirement for optimal growth and Salt 0.3 0.3
development. In this study, the methionine level in basal Lysine 0.01 0.014
diet was only 0.35 and 0.33 %, which was far below the Methionine - -
NRC (1994) recommended level for starter (0.5) and Total 100 100
finisher (0.4%) phases, respectively; indicating that diet Metabolisable energy (ME) 2802 2801
with basal level of methionine could not supply enough Crude protein (CP) 21.8 19.5
methionine and resulted in growth depression and ME:CP 128.5 143.9
supplementation of methionine, provided the limiting amino Lysine 1.19 1.02
acid required for better live weight. Hassan et al. (2003) Methionine .35 0.33
also showed that addition of methionine (0.15%) in broiler Total calcium 1.02 0.93
diets improved the live weight, weight gain of the birds Available phosphorus 0.46 0.36
compared to non-supplemented. Supplementation of
methionine 20% above NRC (1994) requirement has not *Trace mineral premix supplied Mg- 300, Mn- 55, I- 0.4,
Fe- 56, Zn-30 and Cu- 4 mg/kg diet. **The vitamin premix
produced any improvement beyond 100% level indicating
supplied vitamin A- 8,250 IU, vitamin D3- 1,200 ICU; vitamin
excess amino acid was not utilized by birds for protein K- 1 mg; vitamin E- 40 IU; vitamin B1- 2 mg, vitamin B2- 4 mg,
synthesis and muscle development. El-Wafa et al. (2003), vitamin B12-10 mcg; niacin- 60 mg; pantothenic acid-10 mg;
reported that increasing methionine level to 120% of the choline-500 mg/kg diet.

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Table 2. Effect of ASC treatment and methionine levels of diet on live weight (g), weight gain (g), feed intake (g) and feed
conversion ratio at different growth phases

Treatment Live wt Live wt Gain Gain Gain FI/b FI/b FI/b FCR FCR FCR
4th wk 6th wk 0-4 wk 4-6 wk 0-6 wk 0-4 wk 4-6 wk 0-6 wk 0-4 wk 4-6 wk 0-6 wk

Basal ASC0 651 1233 603 583 1186 839 1490 2330 2.02 2.78 2.37
ASC100 656 1221 610 564 1174 829 1502 2331 2.08 2.64 2.34
ASC250 653 1265 606 612 1218 827 1524 2351 1.99 2.51 2.23
100% ASC0 661 1273 614 612 126 833 1505 2338 2.09 2.34 2.21
ASC100 670 1317 621 647 1269 822 1499 2321 1.93 2.43 2.16
ASC250 670 1292 620 622 1244 820 1501 2321 2.02 2.35 2.18
120% ASC0 669 1299 620 630 1250 823 1518 2342 1.99 2.38 2.18
ASC100 666 1303 619 637 1256 823 1521 2344 2.01 2.35 2.17
ASC250 668 1295 621 627 1248 832 1517 2350 2.04 2.33 2.18
SEM 4.42 10.12 4.42 11.34 10.12 3.04 3.92 5.28 0.02 0.03 0.02
ASC
ASC0 659 1274 611 615 1226 832 1504 2336 2.03 2.50 2.25
ASC100 666 1275 618 610 1228 825 1507 2332 2.01 2.47 2.22
ASC250 664 1284 616 620 1237 827 1514 2341 2.02 2.40 2.20
Methionine
Basal 653 1240x 606 587 1193x 832 1505 2337 2.03 2.65x 2.31 x
100% 667 1294y 619 627 1246y 825 1501 2326 2.01 2.37y 2.18 y
120% 668 1299y 620 631 1251y 826 1519 2345 2.01 2.35 y 2.18 y
Probability
ASC NS NS NS NS NS NS NS NS NS NS NS
Met NS P<0.032 NS NS P<0.036 NS NS NS NS P<0.001 P<0.003
ASC × Met NS NS NS NS NS NS NS NS NS NS NS
xy (Met level), Values carrying different superscript in a column vary significantly; NS, nonsignificant.

Table 3. Effect of ASC treatment and methionine levels of diet on carcass weight, organ weight
(% live weight) and meat quality at 6th week of age

Treatment De- Dressed Breast Back Thigh Drumstick Wings Neck pH TBARS WHC
feathered

Basal ASC0 87.44 62.84 24.35 24.35 15.83 15.27 13.49 6.7 5.80 0.154 19.25
ASC100 87.45 62.82 25.02 24.14 15.55 15.05 13.25 6.98 5.82 0.189 18.75
ASC250 87.95 64.57 24.64 24.94 15.10 14.85 1.40 7.06 5.78 0.244 19.00
100% ASC0 88.02 62.40 25.16 24.86 14.91 14.43 13.43 7.42 5.80 0.146 19.00
ASC100 87.36 62.12 24.86 24.45 15.16 14.50 13.46 7.57 5.77 0.176 19.00
ASC250 86.39 62.91 24.72 24.27 15.70 14.15 13.29 7.87 5.80 0.254 19.25
120% ASC0 87.83 65.58 25.19 23.94 15.81 14.27 12.76 8.03 5.82 0.179 20.00
ASC100 86.13 63.43 23.66 24.27 15.16 14.69 14.17 8.05 5.78 0.273 19.75
ASC250 87.93 61.94 24.88 23.76 15.79 14.58 13.57 7.41 5.78 0.261 19.50
SEM 0.24 0.44 0.21 0.21 0.16 0.12 0.11 0.15 0.005 0.15 0.15
ASC
ASC0 87.76 63.60 24.90 24.38 15.52 14.66 13.22 7.39 5.81 0.159l 19.42
ASC100 86.98 62.79 24.51 24.28 15.29 14.75 13.63 7.54 5.79 0.213m 19.17
ASC250 87.42 63.14 24.75 24.32 15.53 14.53 13.42 7.45 5.79 0.253 n 19.25
Methionine
Basal 87.61 63.41 24.67 24.47 15.49 15.06 13.38 6.92x 5.80 0.196 x 19.00
100% 87.26 62.48 24.91 24.52 15.26 14.36 13.39 7.62xy 5.79 0.192 x 19.08
120% 87.29 63.65 24.57 23.99 15.59 14.52 13.50 7.83y 5.79 0.238y 19.75
Probability
ASC NS NS NS NS NS NS NS NS NS P<0.001 NS
Met NS NS NS NS NS NS NS P<0.036 NS P<0.036 NS
ASC × Met NS NS NS NS NS NS NS NS NS NS NS
lmn (ASClevel) xy(Met level), Values carrying different superscript in a column vary significantly; NS, nonsignificant (TBA, mg
malonaldehyde/kg meat sample).

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(Table 3) was not influenced either by ASC treatment or Immune organs and immune status: Statistically, no
methionine supplementation except in case of neck yield, significant variation was observed in either weight of
which increased significantly (P<0.036) and in dose immune organs or immune status (Table 4). But the increase
dependent manner with increased level of methionine. in methionine level produced some appreciable
Methionine is often reported to promote breast yield in improvement in both humoral (HA-SRBC) and cell
broiler chicken (Jankowski et al. 2014) but no such change mediated immunity determined in terms of PHAP response.
was evident in the present trial. Methionine is the most extensively studied amino acid and
Among the meat quality parameters, pH and WHC values it may be required for immunity functions that are distinct
were almost comparable among all the dietary treatments, from transmethylation and trans-sulphuration metabolism,
while TBARS value was influenced by both ASC and so its requirement for immunity appeared to be greater than
methionine level (Table 3). ASC treatment significantly the levels required for growth (Cook 1991). Similarly,
(P<0.001) increased TBARS value in a dose dependent Bunchasak (2009) also reported that the amount of
manner, while methionine level 120% produced a methionine required to support the immune system seems
significant (P<0.036) reduction in TBARS value. One of to be high because it has to be used not only for protein
the main factors determining the oxidative stability of meat synthesis but also to produce some antioxidants. Methionine
is the level of antioxidants deposited during birds’ lifetime supplementation also resulted in significant dose related
(Ahmad and Abbas 2011). As ASC is highly pro-oxidant, it increase in total antibody, immunoglobulin G level and
would have induced some degree of oxidative stress, which response to PHAP, suggesting that methionine was required
in turn consumed most of the antioxidants provided by feed, for some components of the antibody response and might
leaving little for deposition in meat, which can retard be required for thymus derived (T) helper cell function.
oxidative rancidity post-slaughter. Methionine on the other Similarly, Swain and Johri (2000) reported that methionine
hand have antioxidant sparing effect, thus would have supplementation improved leukocyte migration inhibition
resulted in larger deposits of antioxidants like vitamin E, value and enhanced antibody titer of new castle disease
which in turn slowed down the development of meat virus. Thakur and Mandal (2015) reported that when
rancidity. Lohakare et al. (2005) reported that providing feedstuffs like meat-cum-bone meal, sunflower meal and
methionine 20% above the NRC recommended level lower soybean meal were treated with 80 ppm of ASC a small
the TBARS value of breast meat. In addition, Rostango et statistically non-significant reduction in methionine
al. (1995) observed that supplementation of methionine in digestibility was observed for nearly all the feedstuffs tested,
poultry diet tend to reduce fat deposition, hence fewer indicating ASC had a tendency to interact with methionine
substrates were available for development of rancidity. and in turn rendering it less bioavailable. The reduction
might be induced by the action of chlorous acid, active
Table 4. Effect of ASC treatment and methionine levels of diet
on relative weight of immune organs (mg/100g live weight)
component of ASC, on the readily oxidisable functional
and immune status group in methionine rendering it unavailable for absorption.
In the present study, ASC was used at 100 and 250 ppm
Treatment Thymus Bursa Spleen HA PHAP level, which may have further reduced the methionine
bioavailability leading to only a numerical improvement in
Basal ASC0 168.8 107.6 228.2 9.00 28.67 immunity parameters observed.
ASC100 222.3 99.4 184.7 9.17 28.67
It is concluded that feeding ASC treated feed to broiler
ASC250 169.8 98.4 185.0 8.17 30.50
100% ASC0 167.9 100.3 160.5 8.83 33.83
chickens deteriorated the oxidative stability of meat.
ASC100 189.1 79.0 159.0 9.17 34.00 However, methionine supplementation improved broiler
ASC250 175.2 77.5 215.0 9.17 31.00 performance, meat quality and immune-competence and
120% ASC0 156.0 121.9 170.6 10.00 32.17 there is no apparent interaction between ASC and dietary
ASC100 191.3 97.0 221.2 9.83 33.50 methionine levels in broiler feed.
ASC250 194.7 95.7 195.7 9.33 33.67
SEM 0.006 0.004 0.007 0.18 0.97 REFERENCES
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