Effect of Synertox^® on broiler health and performance during aflatoxicosis

A.M. Shareef and E.K.S. Omar

Department of Veterinary Public Health, College of Veterinary Medicine, University of Mosul, Mosul, Iraq

Abstract

Eighty, one-day-old commercial male broiler chicks (Ross 308), were distributed to four dietary treatments with two replicates of 10 chicks each, to study the effect of Synertox^® on broiler performance during aflatoxicosis. Birds were reared for 42 days. All birds were fed on diet with or without aflatoxin (AF) in feed, and with or without Synertox (ST) (enzymes, organic acids and yeast extract) in water. The treatments were as follow: T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST). Body weights and feed intake were recorded weekly. At 42 of bird’s age, five birds were randomly selected for estimation of antibody titres against Newcastle disease (ND; for calculation of the relative weight of bursa of Fabricius, thymus and spleen as well as for scoring liver and kidney lesions. The results show that the total body weight gain through 42 days of rearing period that the gain was highest in T3. In the second order was the weight of the control group (T1), while in the third order was T4. The worst body weight gain was reported in T2, in which chicks fed AF. The same picture in body weight gain was obtained with feed consumption and feed conversion ratio. AF had a significant negative effect on the liver parenchyma of broiler chicks in treatment 2, by changing liver colour from mahogany to enlarged muddy or even to yellowish discolouration, with friable consistency with sub capsular haemorrhages. The addition of Synertox® was effective in restoring the normal red brown liver colour. Kidney were also affected by feeding AF (T2). They were enlarged, swollen and pale in colour. All lymphoid organs tested, thymus, bursa of Fabricius and spleen were negatively affected by feeding AF in broilers at 42 days of age. Synertox® was effective in counteracting the negative effect of AF on relative bursal weight, thymus and spleen relative weights. Aflatoxin had detrimental negative effect on the ND ELISA mean antibody titre. A significant restoring of ND antibody titre to those of control group (T1) was recorded by addition of Synertox® to the drinking water of broilers in (T4) compared with the AF fed T2 group. From the studied parameters it could be concluded that Synertox® could be used in counteracting the negative effects of AF on health and performance of broiler chicks.   

Keywords:Aflatoxin; Synertox; Newcastle disease; Broilers.

Available online at http://www.vetmedmosul.org/ijvs

تأثیر سینرتوکس فی صحة وانتاجیة فروج اللحم المستهلک لسموم الافلا

عقیل محمد شریف و إنعام قاسم سیتو عمر

فرع الصحة العامة البیطریة، کلیة الطب البیطری، جامعة الموصل، الموصل، العراق

 الخلاصة

تمت تربیة 80 فرخا من ذکور أفراخ فروج اللحم نوع (Ross 308) لفترة 42 یوما حیث وزعت الى 4 معاملات وبمکررین بواقع 10 أفراخ لکل مکرر أعطیت الأفراخ علائق حاویة أو خالیة من سموم الافلا فی العلف وحاویة او خالیة من سینرتوکس)المعزز الحیوی الذی یحوی الخمائر والاحماض العضویة ومستخلص الخمیرة) فی الماء وعلى النحو التالی: المعاملة الأولى (صفر سموم الافلا وصفر سینرتوکس)، المعاملة الثانیة (2,5 جزء بالملیون سموم الافلا و صفر سینرتوکس)، المعاملة الثالثة (0,5 مل سینرتوکس لکل لتر ماء وصفر سموم الافلا)، المعاملة الرابعة (2,5 جزء بالملیون سموم الافلا و 0,5 مل سینرتوکس لکل لتر ماء) قدم العلف والماء بصورة حرة طیلة فترة التجربة. تم تسجیل وزن الجسم واستهلاک العلیقة ومعامل التحویل الغذائی أسبوعیا، عند انتهاء التجربة عند 42 یوما جمع الدم لقیاس اضداد مرض نیوکاسل فی مصل الافراخ باستخدام طریقة الادمصاص المناعی المرتبط بالأنزیم (الایلایزا) کما واخذ کل من الطحال وجراب غدة فابریشیا وغدة التوثة لحساب وزانهم النسبیة بالنسبة لوزن جسم الطائر الحی وکذلک تم حساب تدرج الآفة لکل من الکبد والکلیة. اوضحت النتائج ان اعلى زیادة وزنیة خلال فترة التجربة (42) یوما کانت قد سجلت فی المجموعة الثالثة التی دعمت علیقتها بالسینرتوکس فقط، وجاءت مجموعة السیطرة بالمرتبة الثانیة ثم تلتها مجموعة الافراخ التی دعمت علیقتها الملوثة بسموم الافلا بالسینرتوکس. وکانت اقل زیادة وزنیة تعود لمجموعة الافراخ المستهلکة لسموم الافلا فقط وکانت النتائج فی الزیادة الوزنیة مشابهة لنتائج استهلاک العلیقة ومعامل التحویل الغذائی، اثرت سموم الافلا بشکل ملحوض على الکبد وذلک بزیادة حجمه ووزنه و تغیر لونه الى اللون الطینی او الاصفر مع وجود نزف تحت المحفظة وجعله اکثر هشاشه عند مقارنته مع افراخ مجموعة السیطرة، واعاد السینرتوکس حجم الکبد الى الحجم المسجل فی مجموعة السیطرة وکذلک لونه الى الاحمر القهوائی کما وتاثرت الکلیة بسموم الافلا وذلک بشحوبها وتضخمها مقارنة مع مجموعة السیطرة واستطاع السینرتوکس من اعادة لون ووزن الکلیة الى ماهو علیه فی مجموعة السیطرة سجل ضمور فی الغدد اللمفیة التی تمت دراستها وهی غدة فابریشیا والتوثة والطحال جراء استهلاک سموم الافلا عند وزنها بعمر 42 یوم وتمکن السینرتوکس من اعادة الوزن النسبی لهذه الاعضاء الى اوزان مقاربة لمجموعة السیطرة، کان لاستهلاک سموم الافلا تاثیر سلبی ملحوظ على مستویات اضداد مرض نیوکاسل فی افراخ المجموعة الثانیة واستطاع السینرتوکس من تعدیل مستوى الاضداد فی المجموعة الرابعة عن تلک فی المجموعة المستهلکة لسموم الافلا فقط. من مجمل المعاییر المدروسة یمکن القول ان للسینرتوکس فعالیة جیدة فی معادلة التاثیر السلبی لسموم الافلا فی صحة وانتاجیة افراخ فروج اللحم.

Introduction

Aflatoxins (AF), the toxic secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus, are a major concern in the poultry production. AF metabolites are stable and fairly resistant compounds to degradation. These metabolites are usually produced during the growth of the Aspergillus flavus, Aspergillus parasitcus and Aspergillus nominus on certain foods and feedstuffs under favourable conditions of moisture, temperature and aeration (1). Their toxicity depends on several factors including its concentration, the duration of exposure, the species, sex, age, and health status of animals. Contamination of AF in feed causes aflatoxicosis in poultry that is characterised by reduced feed intake, decreased weight gain, poor feed utilization, increased susceptibility to environmental and microbial stresses, and increased mortality (2). AF can also cause productive deterioration which is associated with changes in and impaired immunity, which is able to enhance susceptibility to some environmental and infectious agents (3). Severe economic losses have been reported in the poultry industry due to aflatoxicosis (4). Ultimately, the transmission of AF and its metabolites from feed to animal edible tissues and products, such as liver and eggs, becomes a potential hazard for human health. With increasing knowledge and awareness of AF as a potent source of health hazards to both man and farm animals, producers, researchers and government organizations are making great effort to develop effective preventive management and decontamination technologies to minimise the toxic effects of AF content in foods and feedstuffs.

Appropriate pre and post-harvest contamination can be reduced by using appropriate agricultural practices. However, the contamination is often unavoidable and still remains a serious problem associated with many important agricultural commodities, which emphasizes the need for a suitable process to inactivate the toxin. Besides the preventive management, several approaches have been employed including physical, chemical biological treatments and solvent extraction to detoxify AF in contaminated feeds and feedstuffs (5). The increasing number of reports on detoxification of AF in poultry feed using different techniques has given rise to a demand for practical and economical detoxification procedure. Since the beginning of 1990s, the adsorbent-based studies have been reported to be effective in removing AF from contaminated feed and minimise the toxicity of AF in poultry. Among several adsorbents commercially available in the market, Zeolites (6), bentonites (7) and clinoptilolite (CLI), (8), were preferred because of their high binding capacities for AF and their reducing effect on AF-absorption from the gastrointestinal tract.

Although several mycotoxin detoxitoxifying or adsorbing techniques have been used by the addition of different adsorbents in feed, however, limited information is available on an alternative strategy of aflatoxin detoxification, like dietary acids (9), live microfloral additives (10) and mannanoligosaccharides (11) in diets of chickens may help digestion by inhibiting bacteria growth and regulate pH value in intestines when incorporated into diet formulations. There are many commercial products are used for detoxification including mycotoxin-binding agents holds promise for using contaminated feeds (12). Esterified- glucomannan, a cell wall derivative of Saccharomyces cerevisiae, was protective against aflatoxin B1 and ochratoxin. Lactobaccillus cultures prevented absorption of aflatoxin from intestine (13). This study was carried out as a field trial to evaluate preventive value of detoxifying commercial product mixture including soluble enzymes, yeast extract and organic acids (Synertox) in Ross broilers where health and performance parameters were determined during a period of 6 weeks to evaluate the ameliorating effect of Synertox in broilers during Aflatoxicosis., i.e., by adding Synertox® to water (14).

Synertox® contains an excellent cocktail content of organic acids and their salts, essential micronutrients and the extract of L- form microorganisms, which plays an important role as a chelating agent in the sequestration of the mycotoxin through the gastrointestinal tract, preventing them from absorption, and so help in getting rid of these toxins outside the body.

For understanding the effect of Synertox on aflatoxin detoxification, a study was conducted to investigate its effect on broiler health and performance during aflatoxicosis.

Materials and methods

Birds and diet

Eighty, one-day-old commercial male broiler chicks (ROSS 308), were randomly distributed to four dietary treatments with two replicates of 10 chicks each. During the 42 days, all birds were fed on diet with or without aflatoxin (AF) and Synertox (ST). The ingredient composition of the basal diet is based on NRC (1994) (15), presented in Table 1. Feed and water was available on ad libitum basis. All the birds were vaccinated against Newcastle Disease (N.D.), in drinking water with B1 at 1-day; with oil based vaccine (intra muscular) at day 8 and by Lasota strain at 21 days of age.

Experimental design

The experimental design consists of four dietary treatments; T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST).

Aflatoxin production and analysis

Aflatoxins were produced by the inoculation of fungus on rice as described by (16).

Aflatoxin containing rice was quantitatively evaluated using thin layer chromatography technique according to (17), using the formula:

                       DM X 10⁶

AF mg/kg = ---------------------------

                  E X 200 X 10 X L

The AF containing rice was mixed in feed according to the calculation to get the desirable level of aflatoxin, 2.5 ppm) in the feed. The prepared experimental diets were analysed again using TLC technique to confirm the AF levels.

Detoxification of AF

A commercially available Synertox® plus (NAFA Agriculture), was added in to the water at the recommended dose rate of 0.5 ml/l of drinking water (Treatment 2 and 4).

Table 1. Ingredient and nutrient composition of basal starter and grower diet of broilers.

Sampling

Body weights and feed intake per pen was recorded weekly. At 42 of bird’s age, five birds per replicate were randomly selected for estimation of antibody titres against Newcastle disease (ND). The blood samples (3 ml) were collected from wing vein. The blood serum was separated and analysed by Enzyme linked immune-sorbent assay (ELISA) described by (18). After blood collection, birds were humanely killed and bursa of Fabricius, thymus and spleen were removed and weighed. Score gross lesion of liver and kidney were recorded. For liver, score lesion (0-3): 0: no lesion; 1: congestion; 2: fatty change in liver with muddy or yellowish discolouration; 3: liver yellowish discolouration with sub capsular haemorrhage.. For kidney, score lesion (0-3): 0: no lesion; 1: congestion; 2: paleness discolouration; 3: Paleness with sub capsular haemorrhage.

Statistical analysis

The results (pen means) were subjected to analysis of variance (ANOVA). Means were subjected to Duncan’s test and statistical significance was accepted at P<0.05(19).

Results

Growth performance

The body weight of broiler chicks through 5 weeks of experimental period show significant differences between experimental treatments, Figure 1). There was a significant reduction in body weight in T2 compared with T1. The reduction was noticed from the first week and persist through the following five weeks of rearing broilers. Addition of Synertox ® in T4, was effective in restoring the body weight to that of control one, since there was a significant difference between T4 and T2.

The total body weight gain through 42 days of rearing period are representing in Table (1). From table it is evident that the gain was highest in T3. In the second order was the weight of the control group (T1), while in the third order was T4. The last two treatments (T1 and T4) were not differ significantly in their body weight gain. The worst body weight gain was reported in T2.

The same picture in body weight gain was obtained with feed consumption. A reflection of feed consumption and body weight gain in the four treatment groups was noticed in the results of feed conversion ratio. The lowest feed consumption and feed conversion ratio were reported in T2. Amending Aflatoxin contaminated feed with Synertox® in drinking water was effective in counteracting the negative effect of AF on growth performance parameters of broilers. Addition of Synertox® alone to the drinking water, T3) had not negative effect on the studied growth performance parameters.

Liver and kidney score lesions

The effect of AF and Synertox® on liver and kidney lesion scores in broilers at 42 days of age are presented in table (2). It is evident that AF had a significant negative effect on the liver parenchyma of broiler chicks in treatment 2, when compared with that of control group (T1), by changing liver colour from mahogany (Figure 2), to that which characterised by enlarged muddy or even to yellowish discolouration, with friable consistency and sub capsular haemorrhages, Figure 3). The addition of Synertox® to the drinking water of broilers in treatment 4, was effective in restoring the normal red brown liver colour to that of chicks in treatment 1 (Figure 4). Kidney was also affected by feeding AF (T2), in a significant manner when compared with all other treatment groups (T1, T3 and T4). They were enlarged, swollen and pale in colour.

Relative lymphoid organs weight

All lymphoid organs, thymus, bursa of Fabricius and spleen were negatively affected by feeding AF in broilers at 42 days of age (Table 3). Thymus bursal relative weights were significantly reduced in (T2), compared with control treatment group (T1). Amending drinking water with the detoxifying Synertox ®in treatment 4 was effectively ameliorate the negative AF effect on these organs, compared with T2. Opposite picture was noticed with spleen, since AF had a significant increase in the relative weight of the spleen in T2, compared with (T1, T3 and T4). Detoxifying AF by addition of Synertox to T4 broilers was effective in returning spleen weight to that of control one.

Antibody titre

The means of antibody titre (HA) against Newcastle disease (ND) showed significant (P>0.05) difference between treatments when analysed at 42 day of the trial (Figures 5 and 6). Aflatoxin had detrimental negative effect on the significant (P<0.05) reduction of ND ELISA mean antibody titre of 2441.2 with % CV of 74.92, compared with the control treatment (T1), which had ND titre of 5508.60 with % CV of 36.96. A significant restoring of ND antibody titre to those of control group (T1) was recorded by addition of Synertox® to the drinking water of broilers in (T4) to be 5345 with % CV of 34.42, compared with the AF fed T2 group.

Figure 1: Effect of Aflatoxin and Synertox® on Weekly body weight of broiler chicks*. *Mean body weight for 10 birds. **T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST).

Figure 2: Liver of broiler in treatment 1, fed no Aflatoxin or Synertox®. Normal, red or mahogany liver of broilers at 42 days of age.

Figure 3: Liver of broiler in treatment 2, fed Aflatoxin. Extreme pale, yellowish greasy liver (fatty change), large and fragile of broilers at 42 days of age.

Figure 4: Liver of broiler in treatment 4, fed Aflatoxin and Synertox®. Normal, red brown liver of broilers at 42 days of age.

Figure 5: Newcastle antibody titre of broiler chicks fed no Aflatoxin and Synertox ® at 42 days of age*.

*Mean ELISA titre for 5 birds, **T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST), ***Means with same superscripts do not differ from each other (P>0.05).

Figure 6: Newcastle CV% of antibody titre in broiler chicks fed no Aflatoxin and Synertox ® at 42 days of age*.

*Mean ELISA titre for 5 birds, **T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST)

.

Table 1: Effect of Aflatoxin and Synertox ® on growth performance of broiler chicks at 42 days of age*.

*Mean ±SE for 10 birds, **T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST), ***Means with same superscripts within a column do not differ from each other (P>0.05).

Table 2: Effect of Synertox® ml/L Synertox® (0.5 ml/L of water) on gross score lesions for broiler chicks fed on a diet containing 2.5 ppm aflatoxin (AF) between1 and 42 days of age.

*Mean 10 birds, **T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST), *** The values represent the number of chicks showing changes/number of chicks examined in each treatment group. Severity of the lesions: (-)= no lesions; (±)= mild lesion; (+)=moderate lesion; (++)= sever lesion.

Table 3: Effect of Aflatoxin and Synertox® on relative weight of lymphoid organs of broiler chicks at 42 days of age*.

*Mean ±SE for 5 birds, **T1 (0 ppm AF and no ST); T2 (2.5 ppm AF and no ST); T3 (0 ppm AF and 0.5 ml/l ST); T4 (2.5 ppm AF and 0.5 ml/l ST), ***Means with same superscripts within a column do not differ from each other (P>0.05).