EFFECT OF SACCAHAROMYCES CEREVICIAE (INACTIVATED YEAST) ON QUAIL FED RATION CONTAMINATED WITH FUMONISIN B1 MYCOTOXICOSIS.

M.R. KHALIL; H.A. SHALABY and Z.A.A. KHADER

Animal Health Research Institute, Mansoura Laboratory.

INTRODUCTION

Mycotoxins are secondary toxic metabolites of moulds and their negative effects on poultry production are difficult to overestimate. More than 300 mycotoxins have been shown to induce signs of toxicity in mammalian and avian species. It has been estimated that 25% of the world’s crop production is contaminated with mycotoxins (Fink-Gremmels, 1999) and this number is increasing. These contaminants have a broad range of toxic effects, including carcinogenicity, immunotoxicity and neurotoxicity in addition to reproductive and developmental toxicity (Diana et al., 2006; Elaroussi et al., 2006 and 2008). The most important mycotoxins are aflatoxins, ochratoxin A, fumonisins, trichothecenes and zearalenone. (O’Brien and Dietrich, 2005; Butkeraitis et al., 2004, 2006 and Meissonier et al., 2007).

Fumonisin B-1 (FB-1) is a mycotoxin produced by Fusarium verticillioides, (Loiseau et al., 2007), Fusarium moniliforme-contaminated culture material has been shown to induce toxic responses, including increased mortality, reduced size of the bursa of Fabricius, thymus, and spleen, decreased body weight gain, myocardial degeneration, myocardial hemorrhage, alterations in the hemostatic mechanism, and necrosis of hepatocytes, when fed to chickens, ducklings, and turkey poults (Jeschke et al., 1987; Ogido et al., 2004 and Deshmukh et al., 2007).

Recently, yeast has been successfully used as an effective agent for counteracting aflatoxin or reducing aflatoxicosis in the small intestine (Stanley et al., 1993) and reducing ochratoxin in male broiler chicks (El-Barkouky, 2008). As well, improve the immune response of poultry (Santin et al., 2003). Yildirm and Parlat (2003) showed that, hatchability, fertility and embryonic mortality improved due to adding yeast into laying quail diets.

Therefore the current study aimed to investigate the effectiveness of yeast in countring the harmful effects of fumincin-B1 which was enriched in Japanese quail diets for 9 week of age via performance and physiological parameters.  

MATERIALS and METHODS

A total number of 160 one day old, Japanese quail chicks, obtained from the farm of Biological Application Department, Nuclear Research Center at Inshas were used in the present study. The quail chicks were randomly assigned equally to four groups (40 birds per each). Each group contained 2 replicates of 20 birds. For 9 week experimental period, birds were kept in battery cages, water and feed was provided ad lib. A basal diet of 24% protein and 3000 kcal M.E./kg was formulated to cover all nutrients needed a according to N.R.C 1994, (Table 1). The experimental design was included four tested groups as follows:

1- Group(1): The basal diet and served as control

2- Group(2): The basal diet supplemented with 0.5 % yeast.

3- Group(3): The basal diet supplemented with 0.5 % yeast + 200mg fumincin-      

                      B1( dissolved in distilled water) /kg diet.

4- Group(4): The basal diet supplemented with 200mg fumincin B1/kg diet.

Fumonisin production and analysis.

Fusarium moniliforme culture material, containing FB1, was produced as previously described by Weibking et al. (1995). Culture were grown in sterilized 0.946 liter canning jars containing 100g of whole shelled corn and 100ml of distilled water. A 5ml vial of lyophilized F.moniliforme M-1325, provided by the Fusarium Research Center (Pennsylvania State University Park,Pa.) was added to 500 ml sterilized distilled water and 2ml of this suspension was added to the autoclaved jars. The jars were incubated in the dark in an open room for a total of 5 wk at 27⁰C.

Individual body weight, mortality rate and feed intake were recorded weekly. After sexual maturity at 6 week of old, 12 female and 6 male of each treatment were used. Egg production, fertility, hatchability, embryonic death and weight of hatching chicks were recorded up to 9 weeks of age.

At 6 week old, three birds of each replicate group (6 birds/ group) were randomly slaughtered. Carcass was eviscerated and some organs, including spleen, liver, heart, testes, oviducts and ovaries were removed and weighed individually then their weights were related to live body weight.

Two blood samples were collected during slaughtering the first one was collected with anticoagulant while the second sample was collected without anticoagulant to measure hematological parameters and some serum biochemical parameters. Hematological parameters: Red and White blood cells count (RBC and WBC) were determined according to Natt and Herrick (1952). Hemoglobin concentration (Hb) was determined according to Dacie, and Lewis, (1991). Serum total protein according to Biuret method (Henery, 1964), albumin according to Drupt, (1976). Serum globulin was calculated by subtracting albumins from total protein. Cholesterol was estimated by using the method of Watson, (1960). Liver function tests: alanine amino transaminase (ALT) and aspartate amino transaminase (AST) were determined according to Reitman and Frankel­ (1957). α-phetoprotein (AFP) was determined by radioimmunoassay (RIA) techniques, and used to estimate GPx activity (nmol/min/ml). Uric acid was determined according to James and White (1971) and Createnine was estimated by using the method of Husden and Ropaport (1968), calorimetrically by using commercial kits (bio Merieux - co., Marcyl Etoile – France).

Statistical analysis:

Data were analyzed statistically using the General Linear Models procedure of the program CoStat (1986). The ANOVA model was used according to Snedecor and Cochran (1982) to study the role of Sacchromyces Cervisiae in suppressing the effects of Fumincin mycotoxicosis in Japanese quail. The differences between averages were subjected to Duncan Multiple Range test for all variables (Duncan 1955).

Table 1: Composition and calculated analysis of the basal diets

*Each kilogram of diet contains, A, 12000 I.U; D3, 2000 I.U; E, 10mg; K, 2mg; B1, 1mg; B6,1.5mg;B12, 10µg; B2, 4mg; Pantothonic acid, 10mg;Niacin, 20mg; Folic acid, 1g; Biotin, 50µg; and Choline chloride, 500mg.Copper, 10mg; Iodine, 1mg; Iron, 30mg; Manganese, 55mg; Zing,  55mg and Selenium, 1mg.

** Values were calculated according to the nutrient composition to the NRC (1994).

RESULTS

Body weight, and Mortality rate:

Body weight of birds fed yeast supplemented diet (G2) was significantly higher than that of the control (Table 2) through the experimental periods. The group of birds received fumincin-diet (G4) recorded a significantly lower body weight than all treated groups. Mortality rate of fumincin-diet (G4) and fumincin-yeast group (G3) were higher than other treated groups (Fig 1).

Table 2: Effect of Sacchromyces Cervisiae (Yeast) and Fumonisin on Body weight (gm) in Japanese quail

     ^{a, b, c} Means within row with different superscripts are significantly different (p < 0.05)             

Hatchability, Fertility and Hatching weight:

It is clear that, addition of yeast into basal diet improved egg fertility and hatchability, in contrast enriched FB1 (G4) led to higher depression in both parameters, (Table 3).

Table 3: Effect of Yeast and Fumonisin on Hatchability%, Fertility %, Infertility % and Hatching weight (gm) in Japanese quail

       ^{a, b, c} Means within row with different superscripts are significantly different (p < 0.05)

Egg production percentage:

Egg production percentage of laying quails fed yeast (G2) was significantly higher than the control group (Figure2). Mean egg production % were lower in birds fed Fumonisin (FB1) group 4.

Carcass aspects and internal organs.

There are significant increases in carcass heart, spleen, testis and ovary in group 2 and significant decrease in group 4 (Table 4) compared to the control either in males or females. Relative weight of liver increases (p < 0.05) in the group G4 with fumincinB1 in comparison with the control.

Table 4: Effect of Sacchromyces Cervisiae (Yeast) and Fumonisin on Carcass weight and relative weights of some organs in Japanese quail

^{a, b, c} Means within row with different superscripts are significantly different (p < 0.05)

Some Blood parameters:-

Hematological and Proteins Parameters of the different groups are presented in Table (5). The Red blood cells count, white blood cells count (WBCs) and Hemoglobin value are increased significantly due to adding yeast into diets than control. Also mean values of hemoglobin, white blood cells count, and total erythrocyte count were slightly higher in birds fed FB1. At the same table, the results obtained for total proteins, albumin, globulins and A / G ratio (Table 5) indicated that adding yeast (G2) to the basal diet resulted in a significant increase in total proteins and globulins. G3 come next to G2 in increasing total proteins and globulins, while the control group (G1) showed the significant decrease slightly values. While G4 was increased slightly in quails fed FB 1 than control. Also tables (5) shows significant increase in ALT, AST, cholesterol, uric acid, createnine and alpha feto protein (AFP) and decrease in GPx activity in quails received fumonisin (G4) in comparison with the control group one (G1) and all treatments.

Table 5: Effect of Sacchromyces Cervisiae (Yeast) and Fumonisin B1 on Some Biochemical Parameters in Japanese quail

^{a, b, c} Means within row with different superscripts are significantly different (p < 0.05)

DISCUSSION

Body weight, and Mortality rate:

In the present study showed that body weight of birds fed yeast supplemented diet was significantly higher than that of the control through the experimental periods. The group of birds received fumincin-diet recorded a significantly lower body weight than all treated groups. Mortality rate of fumincin-diet and fumincin-yeast group were higher than other treated groups.

These results were expected due to role of mycotoxin FB1 (higher than 100mg/kg diet) in depressing body weight gain of chicks (Brown et al., 1992; and Henry et al., 2000). Furthermore, Asroni et al. (2006); Elbarkouky, (2008);Girish and Smith, (2008) and Sharmaet al. (2008) observed a reduction in body weight gain of Japanese quails fed fumincin-B1 diet.

Javed et al. (1993) observed an increase in mortality rate of broilers fed FB1-supplemented diets. The beneficial effect of yeast in countering the harmful action of fumincin-B1 has been reported by Abdel-Azeem et al. (2005). In addition Stanley et al. (1993) and Abu-Taleb et al. (2007a) related the beneficial role of yeast due to its structure of cell wall which adsorbed mycotoxin preventing its adverse effect.

Hatchability, Fertility and Hatching weight:

It is clear that, addition of yeast into basal diet improved egg fertility and hatchability, in contrast enriched FB1 led to higher depression in both parameters. The presence of yeast inherent Fumonisin enriched diet success to improve fertility, hatchability of quails than those fed FB1 alone. Yildirm and Parlat (2003)showed that, hatchability, fertility and embryonic mortality are improved due to adding yeast into Japanese quail diets.

Egg production percentage:

Egg production percentage of laying quails fed yeast was significantly higher than the control group. Mean egg production % were lower in birds fed Fumonisin (FB1) group. The improvement in laying performance may be due to the decreasing effect of yeast on pathogenic bacteria, Kim et al. (2002) and Park et al. (2002). Soliman (2003) stated that, including live yeast to laying diets improved egg production and laying performance.

Carcass aspects and internal organs.

The quail chicks fed diets with yeast showed significant increases in carcass and relative wights of heart, spleen, testis and ovary and significant decrease in fed diet contaminated with fumincin B1when compared with the control either in males or females. Relative weight of liver increases (p < 0.05) in the group G4 with fumincinB1 in comparison with the control. Fusarium moniliforme-contaminated culture material has been shown to induce toxic action, including increased mortality, reduced size of the bursa of Fabricius, thymus, and spleen, (Ogido et al., 2004 and Deshmukh et al., 2007). On the other hand, live yeast, Sacchromyces cerevisiae, was found to alleviate the adverse effects of mycotoxins in poultry (Abu Taleb et al., 2007b).

Some Blood parameters:-

The Red blood cells count, white blood cells count (WBCs) and Hemoglobin value are increased significantly due to adding yeast into diets than control. Similar results were recorded by Wakwak     et al. (2003) who observed an increase in WBCs of growing quails fed diets with live yeast. The increment in indicated that, adding yeast into laying quail diets may improve immunity system. Santin et al. (2003) reported that the immune response of birds improved by adding yeast into diets. G4 was increase slightly in quails fed FB 1 than control. Mean values of hemoglobin, white blood cells count, and total erythrocyte count were slightly higher in birds fed FB1. Total leukocyte counts were higher in FB1-fed group because of an increase in the number of heterophils and lymphocytes, Deshmukh et al. (2007) andSharma et al. (2008).

The above mentioned results in this study showed that a significant increase in total proteins and globulins in yeast treated group (G2). While G3 come next to G2 in increasing total proteins and globulins, but the control group (G1) showed the significant decrease slightly values. While G4 was increased slightly in quails fed FB 1 than control. Some authors reported that Japanese quail (Abdel-Azeem et al., 2005), and broiler chicks (Tollba et al., 2004), showed significant increases in the values of total proteins and globulins as the result of feeding Yeast. Sharma et al. (2008). Showed thattotal serum proteins, albumin andglobulins values were higher in all treatment groups of birds fed FB1compared with the control group.

The significant increases in ALT, AST, cholesterol, uric acid, createnine and alpha feto protein (AFP) and decrease in GPx activity in quails received fumonisin (G4) in comparison with the control group one (G1) and all treatments. Yeast observed partial improves the parameters in group three (G3). The previous results agree with those of Sharma,et.al.,(2008) they reported that birds fed FB₁ showed higher values ofcholesterol, aspartate transaminase, lactate dehydrogenase,and creatine kinase than that of the control group.

Glutathione is probably the most abundant natural low molecular weight thiol that is detected in virtually all living cells. It plays a central role in transporting amino acids across cell membranes, catalyzing disulfide exchange reactions, and serving as coenzymes for certain enzymes, detoxifying free radicals and acting as reactive intermediates (Sen and Packer, 1996 and Mohamed, 2000).The liver appears to be a vast reservoir for GSH, it releases from the hepatocytes into bile and blood where it turns over rapidly to be available to other organs (Kaplowitz      et al., 1994).

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