Ameliorative Impact of Mannan Oligosaccharide and Sodium Butyrate on Carcass Traits and Organ Weights During Simultaneous Aflatoxicosis and Ochratoxicosis in Broiler Chickens

Abstract

To study the ameliorative impact of mannan oligosaccharide (MOS) and sodium butyrate (NaB) during simultaneous aflatoxicosis and ochratoxicosis on carcass traits and organ weights, day-old broiler chicks (n=320) were divided into 10 treatment groups (T0: control; T1: T0+300 ppb aflatoxin B1 (AFB1)+250 ppb ochratoxin A (OTA); T2: T1+0.15% MOS; T3: T1+0.3% MOS; T4: T1+500 mg/kg NaB; T5: T1+1000 mg/kg NaB; T6: T1+0.15% MOS+500 mg/kg NaB; T7: T1+0.15% MOS+1000 mg/kg NaB; T8: T1+0.3% MOS+500 mg/kg NaB; T9: T1+0.3% MOS+1000 mg/kg NaB). Each diet was fed to 4 replicated groups of 8 birds each for 42 days. The result showed that shrinkage loss and dressed yield in T1 was lower (P<0.05) than T0. Shrinkage loss and dressed yield in all other treatments did not differ from T1 and T0. The relative values of cut-up parts in T1 were lower (P<0.05) than T0. Inclusion of 0.3% MOS+500 mg/kg NaB to the 300 ppb AFB1+250 ppb OTA contaminated diet (T8) was the most effective combination in ameliorating mycotoxicosis on carcass traits. The relative weights of liver, gizzard, heart, kidney in T0 was lower (P<0.05) than T1. The relative weights of bursa and spleen in T0 was higher (P<0.05) than T1. Addition of 0.3% MOS+500 mg/kg NaB to the 300 ppb AFB1+250 ppb OTA contaminated diet (T8) was the most effective combination in ameliorating adverse effects of mycotoxicosis on organ weights. It was concluded that mycotoxicosis caused by 300 ppb AFB1+250 ppb OTA resulted in decreased shrinkage loss, dressed yield; cut-up parts (thigh, drumstick, breast, back, neck and wing) yield; increase in relative weights of liver, gizzard, heart, kidneys; and decrease in relative weights of bursa and spleen. However, the dietary inclusion of 0.3% MOS+500 mg/kg NaB ameliorated the adverse effects of combine mycotoxicosis as assessed through carcass traits and organ weights in broiler chickens.

Keywords: Aflatoxin B1, Ochratoxin A, Carcass traits, Organ weight, Mannan oligosaccharides, Sodium butyrate, Broiler chicken.

1. Introduction

Mycotoxins frequently contaminate food and animal feeds and thus, pose a potential health threat to various poultry species. It is estimated that around 25% of the world's crops are affected with mycotoxins each year. The toxic effects of mycotoxins i.e. mycoxicosis, is characterized by its nephrotoxic, immunosuppressive, hepatotoxic, carcinogenic, mutagenic and teratogenic effects in animals and poultry (Patil et al., 2005, 2006, 2014, 2017a, b; Katole et al., 2013; Patial et al., 2013; Patel et al., 2015; Patil and Degloorkar, 2016a, b, 2018; Singh, 2019a-f; Singh et al., 2019a, b). Aflatoxin B1 (AFB1) toxicity in poultry causes lowered performance in terms of reduced body weight gain, feed intake and feed efficiency (Silambarasan et al., 2013; Singh et al., 2015a; Singh et al., 2016c), reduced nutrient utilisation (Silambarasan et al., 2013), increased mortality (Khatke et al., 2012b; Sharma et al., 2014), anemia (Singh et al., 2015a; Singh et al., 2016c), hepatotoxicosis and haemorrhage (Churchil et al., 2014; Singh et al., 2015a; Singh et al., 2016c; Pathak et al., 2017), altered biochemistry (Singh and Mandal, 2013; Singh et al., 2013a). Use of mannan oligosaccharide (MOS) and Saccharomyces cerevisiae (SC) (@ 0.05%, 0.1%, 0.2%) alone or their combination moderately ameliorated the adverse effects of 300 ppb AFB1 (Khatke et al., 2012a, b, c). The 0.2% level of MOS and SC was more effective than 0.05% and 0.1% level in counteracting aflatoxin in the feed. MOS appeared to be more efficacious than SC in counteracting aflatoxicosis in broiler chickens. However, the combination of SC and MOS did not show any synergistic effect in counteracting aflatoxicosis (Khatke et al., 2012c). Ochratoxicosis causes a reduction in production performance viz. reduced growth rate, decreased feed consumption, poorer feed conversion (Singh et al., 2016b) and increased mortality (Singh et al., 2015b, 2016b). Ochratoxin contamination at 200 ppb level in broiler diet led to decreased protein, haemoglobin and creatinine, while increased uric acid, alkaline phosphatase, AST and ALT levels in blood. Moreover, ochratoxin impaired both cell mediated and humoral immunity (Singh et al., 2016a). Short chain fatty acids production in the poultry gut supports a healthy gut wall by promoting gut healing and among those short chain fatty acids butyric acid is most important one. MOS also eliminates the mycotoxin from the gut of livestock and poultry by physical adsorption (Zaghini et al., 2005). The objective of the present study was to evaluate the efficacy of MOS and sodium butyrate, and their combination in combating simultaneous toxicity of aflatoxin and ochratoxin in broiler chickens.

2. Materials and Methods

2.1 Production and Analysis of Mycotoxins

Aflatoxin was produced by growing Aspergillus flavus NRRL 6513 on maize. This fungus was obtained from US Department of Agriculture, Illinois, USA. Fermentations were carried out in batches as per the method described by Shotwell et al. (1966). The extraction and estimation of aflatoxin B1 was done as per the procedure of Pons et al. (1966). Aflatoxin contents were finally quantified using a UV spectrophotometer. Ochratoxin was produced as per the method described by Singh et al. (2013b) using Aspergillus westerdijkiae NRRL 3174. The extraction and estimation of ochratoxin A was done as per the procedure of AOAC (1995).

2.2 Experimental Design

Experimental design was completely randomized design. There were ten dietary treatments. Each dietary treatment had 4 replicates and each replicate had 8 chicks. The experiment was conducted in broiler chickens for 6 weeks of age. The various dietary treatments were prepared by mixing the required quantity of mycotoxins, MOS and NaB to get their desired concentration in basal diet (Table 1).

Table 1: Experimental groups and treatments

Ameliorative Impact of Mannan Oligosaccharide and Sodium Butyrate on Carcass Traits and Organ Weights During Simultaneous Aflatoxicosis and Ochratoxicosis in Broiler Chickens - Image 1

2.3 Biological Experiment and Analysis

Day-old broiler chicks were obtained from experimental hatchery, ICAR-Central Avian Research Institute, Izatnagar. The chicks were wing banded, weighed individually and distributed randomly into ten treatment groups on equal weight basis. All birds were reared under standard management conditions and fed with broiler starter ration for 1-21 days and broiler finisher ration from 22 to 42 days. The composition of broiler starter and finisher ration was as below: The starter diet with maize 53.93, soybean meal 42.70, limestone 0.9, dicalcium phosphate 1.70, common salt 1.30, DL-methionine 0.16, TM premix 0.10, vitamin premix 0.15, B complex 0.015, choline chloride 0.05; and finisher diet with maize 63.63, soybean meal 33.40, limestone 0.95, dicalcium phosphate 1.40, common salt 0.20, DL-methionine 0.11, TM premix 0.10, vitamin premix 0.15, B complex 0.015, choline chloride 0.05 were formulated. TM premix supplied Mg, 300; Mn, 55; I, 0.4; Fe, 56; Zn, 30; Cu, 4 mg/kg diet. Vitamin premix supplied Vit A, 8250 IU; Vit. D3, 1200 ICU; Vit. K, 1 mg; vitamin E 40 IU per kg diet. B complex supplied Vit. B1, 2 mg; Vit. B2, 4 mg; Vit. B12, 10 mcg; niacin, 60 mg; pantothenic acid, 10 mg; choline, 500 mg per kg diet. The starter diet contained 22.09% crude protein, 2,848 Kcal ME/kg, calcium 0.99%, available P 0.45%, lysine 1.27%, methionine 0.53% and threonine 1.01%. The corresponding values in finisher diet were 19.04%, 2,946 Kcal/kg, 0.90%, 0.38%, 1.05%, 0.44% and 0.87%. The protein as per AOAC (1995) and calcium contents as per Talapatra et al. (1940) were estimated, while the concentrations of lysine, methionine, threonine, available P and metabolizable energy values were calculated. After 42nd day of age six birds of equal sex from each dietarytreatment were selected as per body weight closer to mean for carcass studies. The birds were kept off feed for eight hours prior to slaughter but drinking water was offered to them. The carcass parameters viz. slaughter traits (shrinkage loss, dressing yield and eviscerated weight); cut-up part yields (breast, thigh, drumsticks, back, neck and wing) and yield of various organs (liver, heart, gizzard, bursa, spleen and kidneys) were recorded and expressed as % live weight.

2.4 Statistical Analysis

All data were statistically analyzed using SPSS software package version 20.0 following one way analysis. All the observations were recorded at 95% (P<0.05) level of significance.

3. Results and Discussion

3.1 Carcass Traits

3.1.1 Slaughter Traits

The data pertaining to shrinkage loss, dressed yield and eviscerated yield were statistically analyzed and presented in Table 2. The shrinkage loss for control group (T0) was 4.24%, which was significantly reduced to 3.04% in mycotoxins fed group (T1). Shrinkage loss in all other treatment groups did not differ significantly from that of T1 and T0. Similar trend of result was observed for dressing yield also, barring T2, which was statistically similar to that of T1. There was no significant difference among various dietary treatments with respect to eviscerated yield. The present study indicated that more shrinkage loss and less dressing yield were seen in mycotoxins alone fed group. However, Silambarsan (2011); Khatke (2012) reported no significant effect on slaughter traits due to aflatoxicosis caused by 300 ppb AFB1 in the diet of broiler chickens. Singh (2015); Singh et al. (2017) also reported no significant effect on shrinkage loss, dressed yield and eviscerated yield due to ochratoxicosis caused by 150 and 200 ppb, respectively of OTA. The present study further revealed that inclusion of MOS and NaB had partial ameliorative effect on these parameters. No significant (P<0.05) change was seen in eviscerated yield due to contamination of mycotoxins (300 ppb AFB1+250 ppb OTA) in the feed. Jahanian et al. (2016) found that the aflatoxin decreased carcass yield and dietary supplementation of MOS upto 2 g/kg resulted in an increase in carcass yield. In contrast to our results, Blair et al. (2004); Konca et al. (2009); Khatke (2012) found no significant effect on relative weights of carcass in MOS supplemented birds. Similarly, Singh (2015) also observed that supplementation of SC had no effect on the slaughter traits of broiler chickens. The information on effect o fNaB supplementation on slaughter traits during mycotoxicosis is lacking in literature.

3.1.2 Cut-Up Parts

The relative values of cut-up parts (thigh, drumstick, breast, back, neck and wing) expressed as percentage of pre-slaughter live weights taken at the end of 6th wk of the trail period in various dietary treatments are presented in Table 3. The relative values of all cut-up parts in control group (T0) was higher (P<0.05), than that of group T1. With regards to thigh, drumstick and back percentage, remaining treatment groups did not differ significantly (P<0.05) from that of T1 and control barring T6 for thigh %, which was significantly similar to that of control and T9 for back %, which was significantly similar to that of group T1. Breast percentage for T2, T6, T7 and T8; and T3, T4, T5 and T9 was statistically similar to that of control (T0); and mycotoxins (300 ppb AFB1+250 ppb OTA) alone fed group (T1), respectively. Percentage of neck parts for groups T2, T3 and T8 was statistically similar to that of control, however, in remaining groups neck % value did not differ significantly (P<0.05) from that of T0 and T1. The percent wing parts of group T8 did not differ significantly (P<0.05) from that of T0 and T1 and remaining groups had significantly similar values to that of mycotoxins fed group (T1).

In the present experiment relative values of cut-up parts (thigh, drumstick, breast, back, neck and wing) were showing lower relative weight in mycotoxins (300 ppb AFB1+250 ppb OTA) fed group (T1) compared to that of control (T0). However, Silambarsan (2011); Khatke (2012) reported no significant effect on cut-up parts due to aflatoxicosis caused by 300 ppb AFB1 in the diet of broiler chickens. Also, Singh (2015); Singh et al. (2017) reported no significant effect on cut-up parts of broilers due to ochratoxicosis caused by 150 and 200 ppb, respectively dietary ochratoxin. The present study revealed that dietary inclusion of 0.15% MOS+500 mg/kg NaB to the mycotoxin contaminated feed significantly improved the relative thigh percentage. MOS, NaB and their different combinations used in experiment partially ameliorated the relative percent values of drumstick, breast, back, neck and wing. In contrast to our study Singh (2015) found supplementation of SC at any level to the ochratoxin (200 ppb) contaminated feed did not produce any significant change in the cut-up parts of broiler chickens. Soltan (2008) found that feeding NaB increased the yield of breast muscle, which suggested that dietary NaB appears to promote anabolism in poultry. This might be the possible cause of NaB salt supplementation improved the cut-up parts values during mycotoxicosis. In the present study, inclusion of MOS and NaB and their different combinations to the 300 ppb AFB1+250 ppb OTA contaminated feed partially ameliorated the adverse effects of mycotoxicosis on relative percent values of drumstick, breast, back, neck and wing. Also, Dietary inclusion of 0.15% MOS+500 mg/kg NaB to the mycotoxin contaminated feed significantly improved the relative thigh percentage.

Table 2: Slaughter traits (% of live weight) as influenced by various dietary treatments

Table 3: Cut-up parts yield (% live weight) of broilers fed different dietary treatments

3.2 Relative Organ

Weight The data pertaining to relative organ weights (liver, gizzard, heart, spleen, bursa, kidney) expressed as percentage live weight were statistically analyzed and are presented in Table 4.

3.2.1 Liver

The relative weights of liver (percent live body weight) was 2.93 in control (T0), which significantly (P<0.05) increased to 3.88 in mycotoxins fed group (T1). The relative weight of liver in groups T2, T3, T4, T6, T7 and T8 was significantly (P<0.05) lower than that of T1 but significantly higher than that of T0, indicating that supplementation of different levels of MOS and NaB to the mycotoxins (300 ppb AFB1+250 ppb OTA) contaminated diet partially ameliorated the adverse effects of mycotoxins on relative liver weights. The relative weights of liver of groups T5 and T9 was statistically similar to that of group T1. In the present study, simultaneous contamination of mycotoxins (300 ppb AFB1+250 ppb OTA) resulted in significant increase in relative weights of liver. This result was in agreement with those of Silambarsan et al. (2015); Khatke (2012) who also reported significant increase in relative liver weights due to aflatoxicosis caused by 300 ppb AFB1 in the diet of broiler chickens. Also, Singh (2015); Singh et al. (2017) reported significant increase in relative liver weights due to ochratoxicosis caused by 150 and 200 ppb dietary ochratoxin, respectively in broiler chickens. Present study indicated that inclusion of different levels of MOS (0.15 and 0.3%) and NaB (500 mg/kg) to the mycotoxin (300 ppb AFB1+250 ppb OTA) contaminated diet partially ameliorated the adverse effects of mycotoxins on relative liver weights but 1000 mg/kg NaB in diet was unable to ameliorate the adverse effect of mycotoxins on relative liver weights in broiler chickens. The cause might be the reduced feed intake in 1000 mg/kg NaB supplementation in the diet of broiler chickens. This result was in agreement with Singh (2015) who reported that supplementation of SC at 0.05 and 0.1% levels ameliorated the adverse effects of OTA on relative weights of liver. Baptista et al. (2002) also found that dehydrated active yeast was able to reduce the hepatotoxicity caused by mycotoxins in hepatocytes. Several researchers confirmatively proved that modified glucomannan, a derivative of yeast cell wall, was able to bind several important mycotoxins (Mahesh and Devegowda, 1996; Volkl and Karlovsky, 1998). In contrast to our study, esterified glucomannan supplementation to the contaminated diet did not significantly diminish the effects of toxins on the relative weights of liver (Arvind et al., 2003). Similarly, Santin et al. (2003) also found that cell wall of SC could not alleviate the adverse effect of ochratoxin on relative weight of liver in broiler chickens. The information on effect of NaB supplementation on relative weights of liver during mycotoxicosis is lacking in literature.

Table 4: Relative weights (% live weight) of organs fed different dietary treatments

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3.2.2 Gizzard

The relative weights of gizzard in the control group (T0), was lower (P<0.05) than that of mycotoxins (300 ppb AFB1+250 ppb OTA) fed group (T1). The relative weights of gizzard of group T8 was statistically similar to that of control. In all other treatment groups the relative weights of gizzard did not differ significantly from that of T1 and T0. In the present study, there was an increase in the relative weights of gizzard due to mycotoxicosis. This result was in agreement with those of Khatke et al. (2012a); Bailey et al. (1998); Kubena et al. (1990); Kubena et al. (1993); Raju and Devegowda (2000) wherein aflatoxicosis caused by 0.3 to 5.0 ppm aflatoxin in feed resulted in increased in relative weights of gizzard. Also, Kubena et al. (1988, 1989); Huff et al. (1992) suggests that the broiler chicks supplemented with OTA at a level of 2 ppm in their diet for 3 weeks of age; showed significant increase in their relative gizzard weights in broiler chicks. Inclusion of 0.3% MOS+500 mg/kg NaB to the mycotoxin (300 ppb AFB1+250 ppb OTA) contaminated diet ameliorated the toxic effect on relative weights of gizzard. Similarly, Khatke et al. (2012a) reported that MOS and SC inclusion in the 300 ppb aflatoxin contaminated diet significantly alleviated the ill effects of aflatoxicosis on relative gizzard weight. However, Santin et al. (2003); Arvind et al. (2003) showed no change on relative weight of gizzard due to supplementation of MOS as cell wall of SC or esterified glucomannan on mycotoxin contaminated diet of broiler chickens.

3.2.3 Heart

The relative weights of heart in control group (T0) was lower (P<0.05) than that of mycotoxins fed group (T1). The relative weights of heart in other treatment groups were statistically similar to that of T1. The relative weights of heart in groups T8 and T3 was higher (P<0.05) than T1 but did not match with that of control group (T0). In present study increased relative heart weights were observed in broiler chickens received mycotoxins (300 ppb AFB1+250 ppb OTA) in feed. Increase in relative heart weight due to aflatoxicosis was earlier reported by several researchers (Bailey et al., 1998; Kubena et al., 1990; Kubena et al., 1993). However, Khatke (2012) reported no significant effect on relative heart weights due to aflatoxicosis caused by 300 ppb AFB1 in the diet of broiler chickens, which could be due to low level of aflatoxin in the diet. Addition of 0.3% MOS alone and along with 500 mg/kg NaB to the mycotoxin contaminated diet partially ameliorated the adverse effects of mycotoxins on relative weights of heart. Stanley et al. (1993); Jahanian et al. (2016) reported that dietary supplementation of SC relieved the deleterious effects of aflatoxin on heart of aflatoxin-challenged broiler chicks. Ghahri et al. (2010), however, showed that inclusion of MOS at the level of 1g/kg had no influence on relative weights of heart in aflatoxin subjected broiler chicks. Also, Khatke et al. (2012a) reported no significant effect on relative heart weights due to inclusion of MOS during aflatoxicosis in the diet of broiler chickens.

3.2.4 Bursa

The relative weights of bursa of Fabricius in control group (T0) was significantly (P<0.05) higher than that of mycotoxins (300 ppb AFB1+250 ppb OTA) fed group (T1). The relative weights of bursa of Fabricius in treatment groups T6, T7 and T8 were statistically similar to that of control (T0). The relative weights of bursa of other groups did not differ (P<0.05) from that of T0 and T1. Results of present study revealed that the chicks received mycotoxins (300 ppb AFB1+250 ppb OTA) alone in diet had lower relative weight of bursa of Fabricius as compared to control birds. This result was in agreement with that of Kubena et al. (1990) wherein decreased bursal weights due to aflatoxicosis caused by 3.5 ppm total aflatoxin were reported in broiler chickens. Silambarsan et al. (2015); Khatke et al. (2012a) also reported that inclusion of 300 ppb AFB1 in the diet of broiler chickens resulted in significant decrease in the relative weights of bursa of Fabricius. Also, Singh et al. (2017); Singh (2015) reported significant decrease in relative bursa weights due to ochratoxicosis caused by 150 and 200 ppb dietary ochratoxin, respectively in broiler chickens. The present study revealed that inclusion of different level combination of MOS (0.15 and 0.3%) and NaB (500 mg/kg) to the mycotoxin (300 ppb AFB1+250 ppb OTA) contaminated diet completely ameliorated the adverse effects of mycotoxins on relative weights of bursa. However, in remaining supplemented groups there was partial amelioration of the adverse effect of mycotoxins on relative weights of bursa in broiler chickens. In agreement with our findings Singh (2015); Singh et al. (2017) concluded that supplementation of 0.1% SC to the 150 and 200 ppb OTA contaminated diet resulted in significant improvement in the relative weights of bursa. Similarly, Bohm et al. (2000); Biagi (2009); El-Barkouky et al. (2010) also reported that addition of SC to the broiler diet provided protection against the adverse effects of OTA on relative weight of bursa.

3.2.5 Spleen

The relative weights of spleen varied between 0.13 (T1) to 0.30% (T0) among various dietary treatments. The relative weights of spleen in groups T8 and T3 was statistically similar to that of control (T0). The relative weights of spleen in other treatment groups was significantly (P<0.05) higher than toxins fed group (T1) but lower (P<0.05) than that of control (T0), barring T5 wherein the relative weights of spleen was statistically similar to that of T1. The present study showed that there was a decrease in the relative spleen weights of broiler chickens received mycotoxins (300 ppb AFB1+250 ppb OTA) in feed. However, increase in relative spleen weights due to aflatoxicosis was earlier reported by several researchers (Bailey et al., 1998; Kubena et al., 1990; Kubena et al., 1993). Silambarsan et al. (2015) reported that inclusion of 300 ppb AFB1 in the diet of broiler chickens resulted in no significant impact on the relative weights of spleen in broilers. Also, Huff and Doerr (1981); Huff et al. (1988); Gibson et al. (1989); Elissalde et al. (1994) indicated that the relative spleen weights increased significantly due to ochratoxicosis caused by 2 to 4 µg/g OTA level in feed for 21 days in broiler chickens.

3.2.6 Kidneys

The relative weights of kidney in control (T0) was lower (P<0.05) than that of mycotoxins (300 ppb AFB1+250 ppb OTA) fed group (T1). The relative weights of kidney in groups T2, T6, T7 and T8 was statistically similar to that of control. The relative weights of kidney in groups T3, T4, T5 and T9 did not differ significantly (P<0.05) from that of control and T1. The study revealed an increase in relative weights of kidney due to simultaneous feeding of aflatoxin and ochratoxin in broiler chickens. Significant increase in relative kidney weights due to aflatoxicosis caused by 0.3 to 5.0 ppm was earlier reported by several researchers (Bailey et al., 1998; Kubena et al., 1990; Kubena et al., 1993; Raju and Devegowda, 2000). Also, several researchers (Huff et al., 1988; Huff et al., 1992; Gibson, 1989; Elissalde et al., 1994) reported increased relative weight of kidney due to ochratoxicosis in broiler chickens. Present study indicated that supplementation of MOS at 0.15% level and its combination with NaB at 500 and 1000 mg/kg levels to the mycotoxin (300 ppb AFB1+250 ppb OTA) contaminated diet ameliorated the adverse effects of mycotoxins on relative weights of kidneys. In contrast, Santin et al. (2003); Arvind et al. (2003) showed no changes on increased relative weights of kidney due to supplementation of MOS as cell wall of Saccharomyces cerevisiae or esterified glucomannan on mycotoxin contaminated diet in broiler chickens.

4. Conclusion

It was concluded that simultaneous aflatoxicosis and ochratoxicosis caused by 300 ppb AFB1+250 ppb OTA resulted in decreased shrinkage loss, dressed yield; cut-up parts (thigh, drumstick, breast, back, neck and wing) yield; increase in relative weights of liver, gizzard, heart, kidneys; and decrease in relative weights of bursa and spleen. However, dietary inclusion of 0.3% MOS+500 mg/kg NaB ameliorated the adverse effects of combine mycotoxicosis as assessed through carcass traits and organ weights in broiler chickens.

This article was originally published in Livestock Research International, April-June, 2019. Volume 07, Issue 02, Pages 116-124.

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Ameliorative Impact of Mannan Oligosaccharide and Sodium Butyrate on Carcass Traits and Organ Weights During Simultaneous Aflatoxicosis and Ochratoxicosis in Broiler Chickens

References

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