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Efficacy of hydrated sodium calcium aluminosilicate in ameliorating ochratoxicosis in broiler chickens

Published: July 26, 2019
By: Ram Singh 1 and A.B. Mandal. / Mycotoxin Laboratory, Division of Avian Nutrition and Feed Technology ICAR-Central Avian Research Institute, Izatnagar - 243 122, Uttar Pradesh, India. 1 Principal Scientist, CIRB, Hisar, India.
Summary

The efficacy of hydrated sodium calcium aluminosilicate (HSCAS) in ochratoxin A (OTA) contaminated diet of broiler chickens was investigated. Day-old chicks (n=240) were divided into six treatment groups (T1 - control; T2 - T1 + 150 ppb OTA; T3 - T1 + 0.25% HSCAS; T4 - T1 + 0.50% HSCAS; T5 - T2 + 0.25% HSCAS; T6 - T2 + 0.50% HSCAS). Each diet was fed to 5 replicated groups of 8 birds each from 0 to 42 days of age. During overall growth period the body weight gain (BWG), feed intake of birds in control group (T1 ) was higher (P<0.05) than those fed ochratoxin (T2 ) or T5 and T6 . The FCR in T1 group was better (P<0.05) than that of T2 , T5 and T6 group. The lower liver and kidney weight, but higher bursa of Fabricius weight was recorded in control group. Higher total serum protein and cholesterol content but lower (P<0.05) uric acid and alkaline phosphatase value was observed in control group (T1) was than that of T2 . Addition of HSCAS at 0.50% level to the ochratoxin contaminated diet partially ameliorated the adverse effects of ochratoxicosis. It was concluded that ochratoxin contamination of feed at the rate of 150 ppb impaired the production performance and altered relative organ weights and blood biochemistry. Incorporation of HSCAS to the ochratoxin contaminated feed could not ameliorate the adverse effects of ochratoxicosis, however, 0.50 percent level of HSCAS partially alleviated the ill effects of ochratoxicosis in broiler chickens.

Key words: Ochratoxin, broiler chicken, performance, HSCAS

INTRODUCTION
The worldwide occurrence of mycotoxins producing fungi and their ability to grow on the variety of economical important cereal crops have rendered them as unavoidable contaminants of human and animal food and feeds. The family of ochratoxins consists of three members known as ochratoxin A, ochratoxin B and ochratoxin C among these ochratoxin A (OTA) is the most toxic one. They are the second major group of mycotoxins characterized after the discovery of aflatoxins. The presence of ochratoxin A, a mycotoxin well known for its nephrotoxic activities, has been reported in finished poultry feed and its ingredients throughout the world (Saleem et al., 2009; Hanif et al., 2006; Zahoor-ul-Hassan et al., 2012). OTA causes significant losses to the poultry industry due to its effects on performance and health. It causes a reduction in growth rate, feed consumption, poor feed conversion ratio and higher mortality (Singh et al., 2015; Singh et al., 2016a; Singh et al., 2016b), suppression of immune function (Verma et al., 2004; Singh, 2015; Singh, 2016b) and impairment of blood coagulation (Raju and Devegowda, 2000). Also, OTA induces degenerative changes and an increase in the weight of the kidney and liver, as well as a decrease in the weights of the lymphoid organs (Elaroussi et al., 2008; Singh et al., 2016a). The addition of naturally occurring inert adsorbents to mycotoxin-contaminated feed has been a popular approach to decrease the toxicity as well as the carryover of mycotoxins from contaminated feed to animal by-products. These adsorbents act by decreasing bioavailability (by adsorption through animal’s gastrointestinal tract) and distribution through the target organs. The present study was aimed to determine whether the addition hydrated sodium calcium aluminosilicate would ameliorate the deleterious effects of ochratoxin A in broiler chicks.
MATERIALS AND METHODS
Ochratoxin production
The lyophilised preparation of Aspergillus westerdijkiae NRRL 3147 was revived on potato dextrose agar medium and used for experimentation. Ochratoxin was produced as per the method described by Singh et al. (2013). Cracked maize (50 g) was taken in 250 ml conical flasks. The moisture content of substrate was adjusted to a level of 35%. Thus flasks were plugged with non-absorbent cotton and sealed with aluminium foil. The flasks were autoclaved for 20 min at 121°C and inoculated with one-week old mycelium of Aspergillus westerdijkiae NRRL 3174. The inoculated flasks were incubated in a BOD incubator for 14 days. After removal from the incubator, the flasks were dried at 70°C and the ochratoxin assays were performed as per AOAC (1995).
Experimental design
Experimental design was completely randomized design (CRD). There were six dietary treatments. Each dietary treatment had 5 replicates and each replicate had 8 chicks. The experiment was conducted in broiler chickens from day-old to 6 weeks of age. The various dietary treatments were prepared by mixing the required quantity of mouldy maize to get the desired concentration of 150 ppb OTA (Table 1). The final concentration of OTA in diet was estimated as per AOAC (1995).
Efficacy of hydrated sodium calcium aluminosilicate in ameliorating ochratoxicosis in broiler chickens - Image 1
Biological experiment and analysis
Day-old broiler chicks (240) were obtained from experimental hatchery. The chicks were wing banded, weighed individually and distributed into 6 groups on equal weight basis. All the birds were reared under standard management conditions from 0–42 days and were fed with broiler starter ration from 1–21 days and broiler finisher ration from 22 to 42 days of age. Weekly individual body weight and feed consumption of each group were recorded. The ingredient and chemical composition of broiler starter and finisher rations were as below (Table 2):
The crude protein as per AOAC (1995) and calcium contents as per Talapatra et al. (1940) were estimated, while the concentrations of lysine, methionine, available P and metabolizable energy values were calculated. Weekly individual body weight and feed consumption of each group were recorded and the FCR was calculated. Mortality was recorded as and when occurred. At the end of sixth week of experimental trial, eight birds per dietary treatment were sacrificed randomly in order to record relative (% of body weight) weights of liver, kidney, spleen and bursa of Fabricius. The blood samples from each treatment group were collected. The serum was separated and stored at -20° C and analyzed for various biochemical parameters using commercial kit manufactured by Span Diagnostics Ltd, SACHIN, Surat.
Statistical analysis
The collected data was subjected to statistical analysis using Software Package for Social Sciences (SPSS Version 16.0). The recorded data were subjected to one-way analysis of variance with comparison among means was made by Duncan‘s multiple range test with significance level of P < 0.05.
Efficacy of hydrated sodium calcium aluminosilicate in ameliorating ochratoxicosis in broiler chickens - Image 2
RESULTS AND DISCUSSION
Body weight gain (BWG)
During starter phase (0-21d), finisher phase (21- 42d), and overall growth phase (0-42d), the BWG of birds in control group (T1 ) was higher (P<0.05) than that of ochratoxin fed group (T2 ) (Table 3). The BWG in T3 and T4 was statistically similar to that of control (T1 ). The BWG in T5 and T6 was statistically similar to that of toxin fed group (T2 ) and lower (P<0.05) than that of control, indicating that addition of HSCAS at any level (0.25 or 0.50%) to the ochratoxin contaminated diet did not ameliorate the adverse effects of ochratoxicosis on body weight gain. The present investigation indicated that addition of 150 ppb ochratoxin to the basal diet of broiler chickens resulted in significant decrease in body weight gain of broilers. Significant reduction in BWG of birds was in agreement with previous investigation with dietary ochratoxin level of 50-100 ppb (Stove et al., 2004; ElBarkouky, 2008; El-Barkouky and Abu- Taleb, 2008), 200 ppb (Sakhare et al., 2007; El-Barkouky et al., 2010; Singh et al., 2015; Singh et al., 2016a ) and 567 ppb (Garcia et al., 2003). Decreased body weight gain might be attributed to the decrease in protein absorption and/ or utilization or to the inhibition of protein synthesis caused by ochratoxin (Kubena et al., 1983, 1988 and 1989). In the present study, addition of HSCAS to the ochratoxin contaminated feed could not ameliorate the adverse effects of ochratoxin on weight gain of broiler chickens. Santin et al. (2002) reported that addition of HSCAS to ochratoxin contaminated feed did not ameliorate the adverse effects of ochratoxins on weight gain in broilers.
Efficacy of hydrated sodium calcium aluminosilicate in ameliorating ochratoxicosis in broiler chickens - Image 3
Feed intake (FI)
During starter phase (0-21d), the feed intake did not differ significantly (P<0.05) among various treatment groups (Table 3). During finisher phase (21-42d) and overall growth period (0-42d), the FI in control group (T1 ) was higher (P<0.05) than that of toxin fed group (T2 ). The FI in group T3 and T4 was statistically similar to that control. The FI in T5 and T6 was statistically similar to that of toxin fed group (T2 ) and lower (P<0.05) than that of control, indicating that addition of HSCAS at any level to the ochratoxin contaminated diet did not ameliorate the adverse effects of ochratoxicosis on feed consumption. In the present study, dietary contamination of ochratoxin resulted in reduced feed consumption in broilers. Similar observations of reduced feed consumption were also reported by earlier workers (Verma et al., 2004; Denli et al., 2008 and Sawale et al., 2009). Other workers (El-Barkouky, 2008; El-Barkouky and Abu-Taleb, 2008; El-Barkouky et al., 2010; Singh et al., 2015; Singh et al., 2016a ), also reported significantly reduced feed intake in broilers fed ochratoxin contaminated feed at a concentration ranging from 50- 200 ppb. In the present study, addition of HSCAS to the 150 ppb ochratoxin contaminated feed did not improve the feed consumption caused by ochratoxin contamination in feed. This result was in agreement with Santin et al. (2002) who reported no effect of HSCAS addition during ochratoxicosis on feed consumption in broilers.
Efficacy of hydrated sodium calcium aluminosilicate in ameliorating ochratoxicosis in broiler chickens - Image 4
Feed conversion ratio (FCR)
With regard to FCR in various growth phases, the FCR during starter phase (0-21d) did not differ significantly among various treatment groups. During finisher phase (21-42d) and overall growth period (0- 42d), the FCR in control group (T1 ) was lower (P<0.05) than that of toxin fed group (T2 ). The FCR in T5 and T6 was statistically similar to that of toxin fed group (T2 ) and higher (P<0.05) than that of control. In the present study, ochratoxin contamination in feed significantly (P<0.05) increased the FCR, thus resulted in poor feed efficiency in broiler chickens. Poor feed efficiency due to ochratoxin contamination in feed was earlier reported by Santin et al. (2006); Hanif et al. (2008); Denli et al. (2008) and Sawale et al. (2009). Singh et al. (2016a ); Sakhare et al. (2007); El-Barkouky (2008); El-Barkouky and Abu-Taleb (2008); El-Barkouky et al. (2010) and Singh et al. (2015) also reported poor feed conversion in broiler chickens fed ochratoxin (50-200 ppb) contaminated feed. The present study revealed that addition of HSCAS to the 150 ppb ochratoxin contaminated feed did not ameliorate the ill effects caused by ochratoxin contamination on feed efficiency. This result was in agreement with Santin et al. (2002).
Organ weight
The average value of relative organ weights (liver, kidney, spleen and bursa of Fabricius) expressed as percentage of live weight were statistically analyzed and presented in Table 4.
Liver: The average value of relative weight of liver (percent of live body weight) in control group (T1 ) was 2.35 which significantly (P<0.05) increased to 3.49 in ochratoxin fed group (T2 ). The relative weight of liver in groups T3 and T4 was statistically similar to that of control. The relative weight of liver in group T5 was statistically similar to that of T2 but higher (P<0.05) than that control. The relative weight of liver in group T6 was lower (P<0.05) than T2 but higher (P<0.05) than that of control. The present study revealed that ochratoxin contamination (150 ppb) in the diet of caused significant (P<0.05) increase in the relative weight of liver. Several researchers have reported a significant increase in the relative weight of liver due to feeding of ochratoxin in broiler chickens (Denli et al., 2008; Hatab, 2003; Elaroussi et al., 2008; Sakhare et al., 2007; El-Barkouky et al., 2010; Hanif et al., 2008; Singh et al., 2015; Singh et al., 2016a). In the present study, addition of HSCAS at 0.5% level to the 150 ppb ochratoxin contaminated feed partially ameliorated the adverse effect of ochratoxicosis on relative weight of liver. Santin et al. (2002) also reported no effect of addition of HSCAS during ochratoxicosis on relative weight of liver in broilers.
Kidney: The average value of relative weight of kidney in control group (T1 ) was lower (P<0.05) than that of ochratoxin fed group (T2 ). The relative weight of kidney in groups T3 and T4 was statistically similar to that of control. The relative weight of kidney in group T5 was statistically similar to that of T2 but higher (P<0.05) than that control. The relative weight of liver in group T6 was lower (P<0.05) than T2 but higher (P<0.05) than that of control. The present study indicated that ochratoxin contamination in the diet caused significant (P<0.05) increase in the relative weight of kidney. Verma et al. (2004) and Hanif et al. (2008) also reported a significant increase in the relative weight of kidney when broilers were fed with OTA at a dietary levels of 0.5, 1.2 and 4.0 ppm over 42 days period. Singh et al. (2015) and ElBarkouky et al. (2010) also reported significant increase in relative weight of kidney at OTA level of 200 µg/kg feed in broiler chickens. In the present study, addition of HSCAS (0.5%) to the ochratoxin contaminated feed partially ameliorated the adverse effect of ochratoxicosis on relative weight of kidney. This result was in agreement with Santin et al. (2002) who reported no effect of addition of HSCAS during ochratoxicosis on feed consumption in broilers.
Spleen: The relative weight of spleen varied between 0.29 to 0.33%. The relative weight of spleen did not vary significantly amongst various dietary treatments. Sreemannarayana et al. (1989), Hashad (1991) and Hanif et al. (2008) also reported that OTA had no effect on the relative spleen weight of broilers fed a diet contaminated with gradual concentration of OTA from 1 to 10 ppm over a four-week period.
Bursa of Fabricius: The relative weight of bursa of Fabricius in control group (T1 ) was 0.28% which reduced (P<0.05) to 0.15% in the ochratoxin fed group (T2). The relative weight of bursa of Fabricius in groups T3 and T4 was statistically similar to that of control. The relative weight of bursa in group T5 was lower (P<0.05) than T1 and statistically similar to that of T2. The relative weight of bursa in group T6 was lower (P<0.05) than T1 but higher (P<0.05) than that of toxin fed group, indicating that addition of HSCAS at 0.5% level to the ochratoxin contaminated feed partially improved the relative weight of bursa. In the present study, 150 ppb ochratoxin contamination in feed caused significant (P<0.05) reduction in the relative weight of bursa. Significant reduction in relative weight of bursa was earlier reported by Stoev et al. (2002); Hanif et al. (2008); Sakhare et al. (2007); Hatab (2003) and Elaroussi et al. (2008). Singh et al. (2016b) and Singh et al. (2015) also reported a significant decrease in the relative weight of bursa due to 200 ppb ochratoxin contamination in feed. The present study revealed that inclusion of HSCAS (0.5%) to the 150 ppb ochratoxin contaminated feed partially alleviated the adverse effects of ochratoxicosis on relative weight of bursa of birds. However, Santin et al. (2002) reported that addition of HSCAS to ochratoxin contaminated feed did not ameliorate the adverse effects of ochratoxins on relative weight of bursa in broilers.
Efficacy of hydrated sodium calcium aluminosilicate in ameliorating ochratoxicosis in broiler chickens - Image 5
Effect on biochemical parameters
The data of various biochemical parameters (total serum protein, cholesterol, uric acid and alkaline phosphatase) was statistically analyzed and the mean values are presented in Table 5.
Total serum protein: The total serum protein content of control group (T1) was higher (P<0.05) than that of ochratoxin fed group (T2 ). The serum protein content in groups T3 and T4 was statistically similar to that of control. The protein content in group T5 was lower (P<0.05) than T1 and statistically similar to that of T2. The protein content in group T6 was lower (P<0.05) than T1 but higher (P<0.05) than that of toxin fed group, indicating that addition of HSCAS at 0.5% level to the ochratoxin contaminated feed partially improved the serum protein content. Ochratoxin (150 ppb) contamination in feed caused significant (P<0.05) reduction in serum protein content. The negative effects of ochratoxin on serum protein in the present study were in agreement with the earlier investigations (Singh et al., 2016b; Stoev et al., 2000; Santin et al., 2002; Hatab, 2003; Elaroussi et al., 2008; El-Barkouky, 2008; ElBarkouky and Abu-Taleb, 2008; Singh et al., 2015). Reduction in serum protein is attributed to the decrease in protein absorption and/or utilisation or to the inhibition of protein synthesis by ochratoxin (Kubena et al., 1983; Kubena et al., 1988; Kubena et al., 1989). Similar decrease in serum protein was also reported when ochratoxin was administered to broiler chickens at 130– 790 ìg/kg (Stoev et al., 2000) and 567 ìg/kg (Garcia et al., 2003). They suggested that low serum protein concentration might have been due to the decrease in albumin and globulin levels or to the degeneration of endoplasmic reticulum that led to pathological changes in the liver that in turn caused a reduction in hepatic protein synthesis, as ochratoxin is known to inhibit hepatic protein synthesis. The present study revealed that inclusion of HSCAS (0.5%) to the 150 ppb ochratoxin contaminated feed partially alleviated the adverse effects of ochratoxicosis on total serum protein content of birds. However, Santin et al. (2002) reported that addition of HSCAS to ochratoxin contaminated feed did not ameliorate the adverse effects of ochratoxins on serum protein in broilers.
Cholesterol: The cholesterol content of control group (T2) was lower (P<0.05) than that of control (T1). The cholesterol content in groups T3 and T4 was statistically similar to that of control. The cholesterol content of group T5 and T6 was statistically similar to that of T2 and lower (P<0.05) than that of T1. The present study revealed significant reduction in cholesterol due to ochratoxin contamination of feed. Schaeffer et al. (1987) and Sreemannarayana et al. (1989) also observed reduction in cholesterol due to ochratoxin contamination in feed of broiler chickens. Inclusion of HSCAS to the 150 ppb ochratoxin contaminated feed did not ameliorate the ill effects of ochratoxicosis on cholesterol content. The effect of addition of HSCAS on cholesterol during ochratoxicosis is lacking in literature.
Serum uric acid: The serum uric acid content of control group (T1 ) was lower (P<0.05) than that of ochratoxin fed group (T2 ). The uric acid content in groups T3 and T4 was statistically similar to that of control. The uric acid value of group T5 and T6 was statistically similar to that of T2 and higher (P<0.05) than that of T1. Ochratoxin contamination resulted in significant (P<0.05) increase in serum uric acid concentration. The significantly increased levels of uric acid in ochratoxin treated birds were in agreement with other reports (Stoev et al., 2000; Ayed et al., 1991; Garcia et al., 2003; Mohiuddin et al., 1993; Patil et al., 2005; Singh et al., 2015; Singh et al., 2016b ). Kubena et al. (1989) indicated that uric acid is the primary product of nitrogen catabolism in chickens and is excreted by the kidney. The elevation in serum uric acid level was accompanied by the increase in kidney weight in the ochratoxin fed birds, indicating impaired renal excretory functions. The present study revealed that inclusion of HSCAS to the ochratoxin contaminated could not alleviate the adverse effects of ochratoxicosis on serum uric acid content of birds. The effect of addition of dietary HSCAS on uric acid during ochratoxicosis is lacking in literature.
Alkaline phosphatase (ALP): The ALP activities in ochratoxin fed group (T2) were higher (P<0.05) than that of control (T1). The ALP value in groups T3 and T4 was statistically similar to that of control. The ALP value in groups T5 and T6 was higher (P<0.05) than T1 and statistically similar to that of T2. In the present study, 150 ppb ochratoxin resulted in increased activities of ALP. Khan et al. (2014) also reported increased activities of ALP due to ochratoxicosis in broiler chickens. Addition of HSCAS at any level to ochratoxin contaminated diet could not ameliorate the adverse effects of ochratoxin on ALP activities. The effect of addition of dietary HSCAS on alkaline phosphatase during ochratoxicosis is lacking in literature.
It was concluded that ochratoxin contamination of feed at the rate of 150 ppb impaired the production performance and altered relative organ weights and blood biochemistry. Incorporation of HSCAS to the ochratoxin (150 ppb) contaminated feed could not ameliorate the adverse effects of ochratoxicosis, however, 0.50 percent level of HSCAS partially alleviated the ill effects of ochratoxicosis on relative organ weights and total serum protein in broiler chickens.
This article was originally published in Indian Journal of Poultry Science (2018) 53(2): 181-187. DOI: 10.5958/0974-8180.2018.00035.1.

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#Minerals in poultry nutrition
#Mycotoxins
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Ram Singh
Ram Singh
5 de septiembre de 2019
Aflatoxins were also estimated, the feed was free from aflatoxins. The objective of the investigation was to induce ochratoxicosis. Dr. Ram Singh Bibyan
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Olga Averkieva
Olga Averkieva
Adisseo
5 de septiembre de 2019
Dear authors, I wonder if you also measured other mycotoxins, especially aflatoxins. The reaction of birds, in my opinion, was more related to symptoms of aflatoxicosis rather than ochratoxicosis. Also the level of ochratoxin A looks not really high to cause significant impact on the performance as well as on some organs.
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