Effects of polyvinylpolypyrrolidone, synthetic zeolite

British Poultry Science (1998) 39: 452–458

Effects of polyvinylpolypyrrolidone,synthetic zeolite and bentonite on serum biochemical and haematologicalcharacters of broiler chickens during a atoxicosis ÆUZ1, V. KURTOG ÆLU2 T. KEC ¸ ECIÇ , H. OG

AND

¨ . DEMET1 O

Departments of Physiology, 1Pharmacology and Toxicology, 2Animal Nutrition and Nutritional Diseases, Faculty of Veterinary Medicine, University of Selçuk, Konya, Turkey Abstract 1. Polyvinylpolypyrrolidone (PVPP), synthetic zeolite (SZ) and bentonite (BNT), when incor-

porated into the diets at 3, 5 and 5 g/kg respectively, were evaluated for their ability to reduce the deleterious effects of 2·5 mg a atoxin (AF; 83·06% AFB1, 12·98% AFB2, 2·84% AFG1, and 1·12% AFG2,)/kg diet on growing broiler chickens from 1 day to 3 weeks of age. A total of 210 chicks were divided into 10 treatment groups (control, AF, PVPP, AF plus PVPP, PVPP plus SZ, AF plus PVPP plus SZ, PVPP plus BNT, AF plus PVPP plus BNT, AF plus SZ and AF plus BNT) each consisting of 21 chicks. 2. When compared to control, the AF treatment signi cantly decreased serum total protein, albumin, inorganic phosphorus, uric acid and total cholesterol, and the values of haematocrit, haemoglobin, mean corpuscular haemoglobin, thrombocyte counts, percentage of lymphocyte and monocyte counts; increased values of white blood cell and heterophil counts. 3. Decreased serum albumin caused by AF was signi cantly ameliorated by PVPP plus BNT. A similar increase was obtained in serum total cholesterol by adding BNT to the AF-containing diet. 4. The serum uric acid, white blood cell counts and percentage of lymphocyte and monocyte counts were numerically intermediate between control and AF values in all of the adsorbent treatments containing AF. The change in thrombocyte counts was alleviated by all of the adsorbent treatments containing AF, except AF plus SZ. The addition of the adsorbents in the AF-free diets did not signi cantly alter the serum biochemical and haematological parameters compared to controls.

INTRODUCTION A atoxicosis is an important disease of livestock and poultry. A atoxins (AF), natural contaminants of foodstuffs, are toxic metabolites produced by Aspergillus avus and Aspergillus parasiticus (Giambrone et al., 1985; Balachandran and Ramakrishnan, 1987). Four types of AF are produced: AFB1, AFB2, AFG1, and AFG2, (Pier, 1992). Prevalence of toxic amounts of AF in foodstuffs and current poultry production methods of con nement rearing on high density diets increase the likelihood that growing chicks may be exposed to AF-contaminated diets (Harvey et al., 1993). Signs of a atoxicosis in poultry include anaemia (Campbell et al., 1983; Keçeci et al., 1995), inhibition of immune function (Thaxton et al., 1974; Campbell et al., 1983; C ¸ elik et al., 1996), hepatotoxicosis (Smith and Hamilton, 1970), mutagenesis, teratogenesis and carcinogenesis (Schull, 1985), anorexia, haemorrhage, poor food utilisation, decreased weight gain and susceptibility to environmental and microbial stresses (Edds and Bortell, 1983). For that reason, AF are a potential threat to poultry health and can cause severe

economic losses in the poultry industries (Hamilton, 1984). Concentrations of serum biochemical (Brugere-Picoux et al., 1987; Kaneko, 1989) and haematological (Huff et al., 1986b; Mani et al., 1993) variables are used to diagnose illness in domestic animals. The AF toxicity in poultry may be manifested by decreased serum concentrations of total protein, albumin, cholesterol, glucose (Harvey et al., 1993), uric acid (Huff et al., 1986b), inorganic phosphorus and calcium (Glahn et al., 1990). Decreased total protein and albumin are consistent indicators of the hepatotoxicity of AF in chickens and turkeys (Huff et al., 1986a; Kubena et al., 1990b, 1991). Furthermore, broiler chicks given 2·5 mg AF/kg diet have shown not only decreased haemoglobin, haematocrit values, thrombocyte counts, percentage of lymphocyte and basophil counts but also an increased percentage of heterophils (Keçeci et al., 1995). Therefore, determination of serum biochemical and haematological changes may help to explain the effects of a atoxicosis in broiler chicks. Removing preformed AF from contaminated foodstuffs remains a major problem. Large-scale,

Correspondence to: T. Keçeci, Department of Physiology, Faculty of Veterinary Medicine, University of Selçuk, 42031 Kampu ¨ s, Konya, Turkey. Accepted for Publication 23rd December 1997. 0007-1668/98/030452-07 $7.00

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1998, British Poultry Science Ltd

EFFECTS OF DIETARY ADSORBENTS ON AFLATOXICOSIS

practical and cost-effective methods for detoxifying AF-containing foodstuffs are not currently available; a variety of physical, chemical and biological methods for detoxifying AF have been employed with limited success (Pasteiner, 1994). One approach has been to use non-nutritive adsorbing materials in the diet in order to bind AF and reduce its absorption from the gastrointestinal tract (Kubena et al., 1990b; Winfree and Allred, 1992; Jindal et al., 1994). The dietary addition of polyvinylpolypyrrolidone (Glavits, 1992; C ¸ elik et al., 1995, 1996; Demet et al., 1996) zeolite (Harvey et al., 1993; Scheideler, 1993), bentonite (Araba and Wyatt, 1991; Winfree and Allred, 1992), hydrated sodium calcium aluminosilicate (Kubena et al., 1990a,b) or activated charcoal (Maryamma et al., 1991; Jindal et al., 1994) have been used to reduce AF toxicity in chickens. Polyvinylpolypyrrolidone (PVPP) added to AF-contaminated diets is reported to prevent effects on serum total cholesterol values, thrombocyte and basophil counts in broiler chicks (Keçeci et al., 1995). It has also been reported that PVPP (3 g/kg diet) addition prevented the broiler chicks from the depressive effects of AF (2·5 mg/kg diet) on peritoneal macrophage functions (C ¸ elik et al., 1996). Scheideler (1993) found that zeolite added to food (5 g/kg) ameliorated the deleterious effects of AF (2·5 mg/kg food) on serum calcium and inorganic phosphorus concentrations. Zeolitic compound (5 g mordenite/kg), when added to AFcontaminated (3·5 mg/kg food) diets, diminished the AF-related changes in serum uric acid and albumin in broiler chicks but changes in the serum inorganic phosphorus, calcium, total protein and cholesterol were unaffected by the presence of modenite (Harvey et al., 1993). Hydrated sodium calcium aluminosilicate (HSCAS) at a concentration of 5 g/kg diet signi cantly reduced many of the deleterious effects caused by AF in chickens (Kubena et al., 1990a,b, 1993a,b). Winfree and Allred (1992) reported that bentonite (BNT) reduced measurable AFB1 in sh food. In addition, Araba and Wyatt (1991) indicated that decreased weight gains and poor food utilisation of the growing broiler chicks given AF (5 mg/kg food) were signi cantly improved by the addition of 10 g sodium BNT/kg diet. The present study was undertaken to determine the effects of PVPP, synthetic zeolite (SZ) and BNT in reducing the AF toxicity in broiler chicks by observing their effects on various serum biochemical components and haematological features.

MATERIALS AND METHODS Two hundred and ten day-old unvaccinated broiler chicks Avian of both sexes were obtained from a commercial hatchery. Individually weighed chicks were divided at random into 10 groups, each of 21

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chicks. The chickens were housed in electrically heated batteries under continuous uorescent lighting where food (maize and soyabean meal diet, 220 g protein and 12·97 MJ ME/kg; without added antibiotics, coccidiostats, or growth promoters) and water were available ad libitum from 1 to 21 d. The food for all 10 groups was tested for possible residual AF before feeding (Howel and Taylor, 1981) and was found negative. The experiment consisted of 10 dietary treatments: Control, normal diet; 2·5 mg AF/kg food; 3 g/kg PVPP(p-6755, Sigma); 2·5 mg AF/kg food plus 3 g/kg PVPP; 3 g/kg PVPP plus 5 g/kg SZ; (Z-3125, Sigma); 2·5 mg AF/kg food plus 3 g/kg PVPP plus 5 g/kg SZ; 3 g/kg PVPP plus 5 g/kg BNT (B-3378, Sigma); 2·5 mg AF/kg food plus 3 g/kg PVPP plus 5 g/kg BNT; 2·5 mg AF/kg food plus 5 g/kg SZ; 2·5 mg AF/kg food plus 5 g/kg BNT. The broilers were maintained on these treatments to 3 weeks of age. The AF was produced by fermentation of rice by Aspergillus parasiticus NRRL 2999 from USDA, Peoria Illinois (Shotwell et al., 1966; Demet et al., 1995). The fermented rice was autoclaved, dried and ground to a powder. The AF content was analysed by the method of Shotwell et al., (1966) and measured using a uorescence spectrophotometer (Perkin Elmer MPF 43-A with detection limit: 0·1 m g AF/kg rice powder; recovery of the extraction method: 92%). The AF within the rice powder consisted of 83·06% AFB1, 12·98% AFB2, 2·84% AFG1 and 1·12% AFG2 based on total AF in the rice powder. The rice powder was incorporated into the basal diet to provide the desired amount (2·5 mg/kg) of AF in the kg food. The diet containing AF alone was analysed and the presence of parent AF was con rmed spectrophotometrically. When the chicks reached 3 weeks of age, the feeding trial was terminated and 10 broilers from each treatment were selected at random and bled by cardiac puncture. Serum concentrations of total protein, albumin, inorganic phosphorus, calcium, uric acid, total cholesterol and glucose were determined on a clinical chemistry analyzer (Gilford Impact 400E, Gilford Systems, Ohio). The red blood cell (RBC), white blood cell (WBC) and thrombocyte counts were determined by a haemocytometer method using Natt-Herrick solution; hematocrit values, haemoglobin (HBG) amounts and erythrocyte sedimentation rates were measured by microhaematocrit, cyanmethaemoglobin and Wastergreen methods respectively; the mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) were calculated; differential leukocyte counts were determined as described by Konuk (1975). Data for serum biochemical and haematological values were grouped and expressed as mean 6 pooled standard errors of the means.

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Then, the results obtained were statistically analysed using Duncan’s multiple range test (Inal, 1992). Statements of statistical signi cance are based on P , 0·05. RESULTS Data presented in Table 1 show the effects of dietary treatments on serum biochemical values of broilers at 3 weeks of age. Feeding AF alone caused signi cant decreases in the serum total protein, albumin, inorganic phosphorus, uric acid and total cholesterol. Adding PVPP plus BNT to the diet containing AF caused a signi cant improvement in serum albumin. Similar amelioration was found in serum total cholesterol by adding BNT alone to AF-containing diet. Serum uric acid was numerically intermediate between control and AF values in all of the adsorbent treatments containing AF. None of the treatments altered the serum concentrations of glucose and calcium. The effects of treatment on haematological values of broilers are given in Tables 2 and 3. The chicks given AF alone showed a signi cant decrease in haematocrit, HBG, MHC, thrombocyte counts, percentage of lymphocyte and monocyte counts. In addition, WBC and percentage of heterophil counts increased in the chicks given AF alone compared to the control chicks. There were no signi cant differences in RBC, MVC, MCHC, erythrocyte sedimentation rates, percentage of basophil and eosinophil counts with these treatments. A signi cant improvement was observed in the thrombocyte counts by all of the adsorbent treatments containing AF, except AF plus SZ. The AF-related changes in the other haematological parameters were statistically unaffected by adding PVPP, SZ, BNT and their combinations to the AF-containing diet. However, the WBC counts, and lymphocyte and monocyte fractions were slightly ameliorated in all of the adsorbent treatments containing AF. The addition of the adsorbent in the AF-free diets did not signi cantly alter the serum biochemical and haematological parameters compared to controls. DISCUSSION Mycotoxins, particularly AF, have been reported to produce severe economic losses and health problems in the poultry industry (Hamilton et al., 1977; Phillips et al., 1988; S¸anli, 1995). Because AF are produced by ubiquitous fungi, their presence in poultry diets is not unexpected. Diagnosis is rather dif cult and medical treatment may be almost impossible in a atoxicosis cases of poultry. Chronic a atoxicosis may be diagnosed by determining the serum biochemical and haematological alterations before clinical symptoms become apparent. Even small amounts of AF are dangerous for animal

health because of its detrimental effect on some biochemical (Schell et al., 1993; Fernandez et al., 1994) and haematological values (Campbell et al., 1983; Mohiuddin et al., 1986; Mani et al., 1993). In this study, experimental a atoxicosis was induced in day-old broilers by feeding 2·5 mg total AF/kg diet for 3 weeks (Campbell et al., 1983; Huff et al., 1986b, 1988; Kubena et al., 1993a). Agents which act as antidotes or antagonise the effects of AF have therapeutic and economic importance. PVPP, SZ and BNT were selected for use in the present study. We considered that toxin absorption from the gastrointestinal tract can be prevented by formation of a complex with AF (Theng, 1974; Interpremix GmbH., 1991; Harvey et al., 1993). When compared to controls, the addition of PVPP, SZ, BNT and their combinations in the diet without AF have not signi cantly altered the serum biochemical and haematological parameters of growing broilers. These data show that these adsorbents in this study were inert and non-toxic. In intoxicated broilers, decreases in serum total protein and albumin concentrations were found (P , 0·05), in agreement with ndings in approximately 21-d-old chicks (Campbell et al., 1983; Kubena et al., 1990b, 1993a). AF inhibits DNA-dependent RNA polymerase and causes impairment of nuclear DNA template function, resulting in general inhibition of protein synthesis (Gelbolin et al., 1966; Yu, 1977). As a result, hypoproteinaemia is a common effect of a atoxicosis (Huff et al., 1986a, b). Kubena et al. (1990a, b) reported that decreased serum total protein and albumin in broilers as the results of AF were not alleviated by HSCAS. In the present study, serum total protein was reduced by all of the treatments containing AF, but adding PVPP plus BNT to the diet containing AF caused a signi cant improvement in serum albumin (P , 0·05). AF may cause alteration of calcium and inorganic phosphorus metabolism. It may directly alter the renal, intestine and parathyroid regulation of calcium and inorganic phosphorus (Glahn et al., 1991). Decreases seen in serum inorganic phosphorus agree with the ndings of other studies (Huff et al., 1986b; Glahn et al., 1990; Kubena et al., 1990a). However, there was no signi cant difference in serum calcium with either AF or other additions and decreased inorganic phosphorus of chicks given AF was not alleviated by treatment with adsorbent (P . 0·05). A signi cant decrease in serum uric acid was seen in chicks given AF alone (P , 0·05) as reported by Harvey et al. (1993) and Huff et al. (1986b). In the present study, the uric acid in all the adsorbent treatments containing AF were close to those seen with the control diet. Our report agrees with others showing that the addition of 5 g/kg zeolite (Harvey et al., 1993) or 3 g/kg PVPP (Keçeci et al., 1995) caused uric acid to return to control values in growing broilers given

1

2

2

2

1

1

1

1

1

2

1

2

2

1

1

2

2

2

2

SZ

1

2

1

1

2

2

2

2

2

2

BNT

2·07 6 1·20 6 2·04 6 1·15 6 1·90 6 1·19 6 1·99 6 1·25 6 1·08 6 1·28 6

0·12a 0·12b 0·10a 0·10b 0·09a 0·07b 0·13a 0·10b 0·06b 0·13b

Total protein (g/dL) 1·24 6 0·90 6 1·26 6 1·01 6 1·20 6 0·95 6 1·21 6 1·10 6 0·96 6 1·06 6 0·05a 0·32d 0·05a 0·06cd 0·07ab 0·05cd 0·06ab 0·07abc 0·05cd 0·06bcd

Albumin (g/dL) 6·17 6 5·44 6 6·16 6 5·41 6 6·14 6 5·36 6 6·13 6 5·58 6 5·53 6 5·44 6 0·08a 0·12b 0·13a 0·23b 0·16a 0·12b 0·16a 0·14b 0·13b 0·13b

Inorganic phosphorus (mg/dL) 9·60 6 9·29 6 9·54 6 9·25 6 9·52 6 9·22 6 9·50 6 9·36 6 9·20 6 9·29 6 0·06a 0·09a 0·12a 0·10a 0·11a 0·10a 0·12a 0·11a 0·11a 0·10a

Calcium (mg/dL) 6·27 6 5·29 6 6·11 6 5·66 6 6·15 6 5·52 6 6·11 6 5·79 6 5·50 6 5·79 6 0·12a 0·36b 0·16a 0·28ab 0·17a 0·28ab 0·29a 0·27ab 0·25ab 0·36ab

Uric acid (mg/dL) 135·10 6 118·30 6 132·38 6 124·70 6 130·11 6 118·25 6 132·84 6 125·60 6 117·41 6 126·60 6 1·90a 3·59de 2·56abc 2·03cde 1·77abc 3·41de 2·25ab 3·20bcd 3·26e 1·73bc

Total cholesterol (mg/dL) 159·44 6 152·30 6 159·75 6 158·49 6 157·18 6 154·17 6 158·35 6 155·40 6 153·40 6 156·37 6 1·68a 3·77a 2·30a 2·85a 2·45a 2·08a 2·28a 2·97a 3·29a 2·77a

Glucose (mg/dL)

1

2

2

2

1

1

1

1

1

2

1

2

2

1

1

2

2

2

2

SZ

1

2

1

1

2

2

2

2

2

2

BNT

2·30 6 1·89 6 2·30 6 2·08 6 2·26 6 2·01 6 2·29 6 2·09 6 1·96 6 2·13 6

0·17a 0·12a 0·20a 0·18a 0·17a 0·19a 0·19a 0·16a 0·18a 0·17a

RBC4 (106 mm3 )

31·9 6 25·7 6 31·6 6 28·2 6 31·0 6 26·5 6 31·5 6 28·5 6 26·2 6 28·8 6

1·55a 1·30c 1·89a 1·66abc 1·67ab 0·32bc 1·70a 1·54abc 1·88bc 1·43abc

Haematocrit (%) 140·89 6 137·09 6 140·77 6 138·57 6 139·47 6 134·70 6 140·92 6 139·67 6 138·98 6 138·28 6 3·56a 4·53a 4·48a 4·78a 3·73a 5·08a 5·04a 8·26a 4·52a 4·22a

MCV5 (m m3) 7·79 6 6·16 6 7·69 6 6·77 6 7·44 6 6·60 6 7·52 6 6·80 6 6·28 6 6·91 6 0·46a 0·44c 0·56ab 0·52abc 0·49abc 0·51abc 0·53abc 0·45abc 0·50bc 0·48abc

Haemoglobin (g/100 ml) 34·16 6 32·63 6 34·10 6 32·79 6 33·92 6 33·56 6 34·07 6 32·71 6 32·30 6 32·65 6

0·44a 0·34bc 0·57ab 0·31abc 0·38ab 1·12abc 0·44ab 0·28abc 0·33c 0·28abc

MCH6 (pg)

26·72 6 26·00 6 26·17 6 25·85 6 25·87 6 27·05 6 25·71 6 25·92 6 26·74 6 25·85 6

0·64a 0·96a 0·39a 0·62a 0·40a 0·99a 0·53a 0·94a 2·12a 0·60a

MCHC7 (%)

28·9 6 33·5 6 30·5 6 32·5 6 31·9 6 32·9 6 31·2 6 32·2 6 33·1 6 32·0 6

1·48a 1·41a 1·69a 1·86a 1·68a 1·58a 1·70a 1·58a 1·73a 1·66a

1st hour

1

a-c

57·4 6 62·4 6 57·9 6 60·8 6 59·0 6 61·0 6 58·5 6 59·6 6 61·8 6 59·1 6

3·26a 3·01a 3·54a 3·96a 3·52a 3·24a 3·62a 3·28a 3·52a 3·44a

2nd hour

121·1 6 131·7 6 122·1 6 128·4 6 124·3 6 128·8 6 123·7 6 125·7 6 130·5 6 124·7 6

6·88a 6·40a 7·46a 8·35a 7·44a 6·83a 7·61a 6·91a 7·41a 7·23a

24th hour

Erythrocyte Sedimentation Rate (mm in fall; 45°)

Values within columns with no common superscripts are signi cantly different (P , 0·05), according Duncan’s multiple range tests. 3 g/kg diet; 2,3 5 g/kg diet; 4Red blood cells; 5Mean corpuscular volume; 6Mean corpuscular haemoglobin; 7Mean corpuscular haemoglobin concentration. *Values represent the x¯ 6 SEM of 10 groups of 10 broiler chicks each per treatment.

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PVPP

AF

Treatment

Table 2. Effects of polyvinylpolypyrrolidone (PVPP)1, synthetic zeolite (SZ)2 and bentonite (BNT)3 on some haematological parameters for broiler chicks fed on diets containing 2·5 mg total aatoxin (AF)/kg diet at 21 days of age*

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a–e

Values within columns with no common superscripts are signi cantly different (P , 0·05), according Duncan’s multiple range tests. 3 g/kg diet; 2,35 g/kg diet. *Values represent the x¯ 6 SEM of 10 groups of 10 broiler chicks each per treatment.

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1

PVPP

AF

Treatment

diet at 21 days of age*

Table 1. Effects of polyvinylpolypyrrolidone (PVPP)1, synthetic zeolite (SZ)2 and bentonite (BNT)3 on serum biochemical values for broiler chicks fed on diets containing 2·5 mg total aatoxin (AF)/kg


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1

2

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2

1

1

1

1

1

2

1

2

2

1

1

2

2

2

2

SZ

1

2

1

1

2

2

2

2

2

2

BNT

0·30 6 0·20 6 0·32 6 0·28 6 0·31 6 0·25 6 0·33 6 0·27 6 0·22 6 0·28 6 0·1abc 0·02f 0·02ab 0·01bcd 0·02abc 0·01de 0·02a 0·01cd 0·02ef 0·01bcd

Thrombocyte ( 3 105 /mm3 ) 35·74 6 41·37 6 34·54 6 38·63 6 34·30 6 39·21 6 35·28 6 38·26 6 39·43 6 37·25 6

(3 0·93bcd 1·51a 1·28cd 1·03abc 0·93d 2·00ab 1·90bcd 1·29abcd 1·09ab 1·76abcd

WBC 103 /mm3 ) 33·7 6 47·0 6 34·6 6 42·9 6 35·4 6 42·3 6 33·6 6 40·3 6 40·3 6 38·8 6 2·58c 2·91a 2·27bc 2·75ab 3·49bc 2·12abc 2·76c 2·08abc 4·25abc 4·35abc

Heterophil 58·9 6 47·8 6 58·1 6 51·0 6 57·4 6 51·7 6 59·1 6 53·3 6 53·7 6 54·5 6 2·53a 2·76b 2·40a 2·67ab 3·55a 1·89ab 2·89a 1·84ab 4·07ab 4·51ab

Lymphocyte 3·1 6 2·0 6 3·2 6 2·3 6 3·0 6 2·4 6 2·9 6 2·5 6 2·5 6 2·7 6 0·38a 0·30b 0·36a 0·42ab 0·26ab 0·34ab 0·31ab 0·27ab 0·34ab 0·37ab

Monocyte 2·7 6 2·1 6 2·6 6 2·4 6 2·7 6 2·3 6 2·8 6 2·5 6 2·3 6 2·5 6

1

a–f

0·26a 0·23a 0·31a 0·27a 0·30a 0·37a 0·36a 0·31a 0·30a 0·31a

Basophil

Differential leukocyte counts (%)

Values within columns with no common superscripts are signi cantly different (P , 0·05), according Duncan’s multiple range tests. 3 g/kg diet; 2,3 5 g/kg diet. *Values represent the x¯ 6 SEM of 10 groups of 10 broiler chicks each per treatment.

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1

1

2

1

2

1

2

2

2

1

PVPP

AF

Treatment

chicks fed on diets containing 2·5 mg total aatoxin (AF)/kg diet at 21 days of age*

1·6 6 1·1 6 1·5 6 1·4 6 1·5 6 1·3 6 1·6 6 1·4 6 1·2 6 1·5 6

0·16a 0·10a 0·31a 0·16a 0·17a 0·15a 0·31a 0·22a 0·13a 0·17a

Eosinophil

Table 3. Effects of polyvinylpolypyrrolidone (PVPP)1, synthetic zeolite (SZ)2 and bentonite (BNT)3 on thrombocyte, total leukocyte (WBC) and differential leukocyte counts for broiler

456 T. KEC ¸ ECIÇ ET AL.


AF. In this study, decreased serum cholesterol in chicks given AF (2·5 mg/kg diet) is consistent with the general reduction of lipogenesis (Donaldson et al., 1972) and impaired lipid transport (Tung et al., 1972) in chicks and speci c inhibition of hepatic cholesterol biosynthesis (Kato et al., 1969) in rats by AF. Serum pro les of chicks given AF plus SZ were similar to those obtained by Harvey et al. (1993) who reported that decreased serum inorganic phosphorus, total protein and cholesterol were not ameliorated by zeolitic compounds in broiler chicks. Whereas in the present study decreased serum total cholesterol in growing broilers were improved by adding 5 g/kg BNT alone with the AF. AF might have affected the tissues of haemopoietic (Campbell et al., 1983; Mohiuddin et al., 1986) and immune systems (Thaxton et al., 1974; Campbell et al., 1983; OgÆuz, 1997) and thereby the production of the cells might have been affected. Various studies have reported that the haematocrit, HBG (Campbell et al., 1983; Keçeci et al., 1995), MCV (Huff et al., 1986b), RBC counts (Huff et al., 1988) and thrombocyte counts were decreased by AF and a atoxicosis caused lymphocytopenia and heterophilia (Keçeci et al., 1995) in growing broilers. In this study, the decreases in the mean values of haematocrit, HBG, MCH, thrombocyte counts in AF-fed chicks indicate the depressing effect of AF on haemopoietic tissue. The decreases in haematological values in the chicks given AF may be related to the inhibition of protein synthesis by AF (Kubena et al., 1993b). It has been reported that AF caused depletion of lymphoid cells in the bursa of Fabricius, thymus and spleen in broiler chicks (Dafalla et al., 1987; Ram et al., 1988; OgÆuz, 1997; Kiran et al., 1998). These support our results showing that a atoxicosis caused lymphocytopenia and monocytopenia in chicks given AF alone. There was a signi cant increase in WBC counts mainly consisting of heterophils in chicks given AF (P , 0·05). These increases in WBC and percentage of heterophil counts suggest that the toxin is eliciting an in ammatory response in the chicks. These data agree with the report by Mohiuddin et al. (1986). Keçeci et al. (1995) reported that decreased thrombocyte and basophil counts from AF (2·5 mg/kg food) in 3-week old broilers were ameliorated by 3 g/kg PVPP. It has been reported that 3 g/kg PVPP addition protected the broiler chicks from the depressive effects caused by AF (2·5 mg/ kg diet) on peritoneal macrophage functions (C ¸ elik et al., 1996). In addition, BNT or BNT plus PVPP addition ameliorated pathological defects of the liver, bursa of Fabricius and thymus in growing broilers given 2·5 mg AF/kg food (OgÆuz, 1997). In the present study, a signi cant improvement was observed in the thrombocyte counts by all of the adsorbent treatments containing AF (P , 0·05), except SZ. There was an intermediate alleviation in the alterations of haematological parameters except the heterophil values by feeding 3 g/kg PVPP in

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combination with AF when compared to control values. Similar results were observed for the HBG, MCH, WBC counts, percentage of heterophil, lymphocyte and monocyte counts by PVPP plus SZ in broilers given AF. However, changes in haematocrit, haemoglobin, MHC and thrombocyte counts caused by AF were unaffected by the presence of SZ (P . 0·05). Winfree and Allred, (1992) reported that BNT reduced measurable AFB1 in sh diets. These data support the ndings that haematocrit, HBG, MCH, lymphocyte and monocyte were slightly increased and that the WBC count and percentage of heterophil count were slightly decreased by the addition of 5 g/kg BNT or 3 g/kg PVPP plus 5 g/kg BNT in the chicks given AF. This study has shown that some serum biochemical and haematological changes could be ameliorated by the administration of PVPP and BNT to the diet at doses of 3 and 5 g/kg respectively in broiler chicks given 2·5 mg AF/kg diet. PVPP and BNT were more effective than SZ in this study. Although the protective effect of the adsorbents used in this study against the toxic effects of AF was not as great as might have been predicted, these improvements should help solve the AF problem in chicks. REFERENCES ARABA, M. & WYATT, R.D. (1991) Effects of sodium bentonite, hydrated sodium aluminosilicate (NovaSilTM) and ethacal on a atoxicosis in broiler chickens. Poultry Science, 70: 6(Abstract). BALACHANDRAN , C. & RAMAKRISHNAN , R. (1987) An experimental study on the pathology of a atoxicosis in broiler chicken. Indian Veterinary Journal, 64: 911–916. BRUGERE-PICOUX, J., BRUGERE, H., BASSET, I., SAYED, N., VAAST, J. & MICHOUX, J.M. (1987) Biochimie clinique en pathologie aviaire. Interet et limites des dosages enzymatiques chez b poule. Recuel Medicine Vetereneri, 163: 1091–1099. CAMPBELL, M.L., MAY, J.D., HUFF, W.E. & DOERR, J.A. (1983) Evaluation of immunity of young broiler chickens during simultaneous a atoxicosis and ochratoxicosis. Poultry Science, 62: 2138–2144. ¨ ., DO¨NMEZ, H.H., OGÆUZ, H. & BOYDAK , M. Ç ELIK, I., DEMET, O (1996) Determination of phagocytic and candidacidal activities of peritoneal macrophages isolated from chickens fed with a atoxin and an a atoxin adsorbing agent, polyvinylpolypyrrolidone. Journal of Veterinary Science, 12: 145–151. DAFALLA , R., YAGI, A.I. & A DAM, S.E.I. (1987) Experimentally a atoxicosis in hybro type chicks: Sequential changes in growth and serum constituents and histopathological changes. Veterinary and Human Toxicology, 29: 222–226. ¨ ., OGÆUZ, H., Ç ELIK, I. & ADIGU¨ZEL, H. (1995) ProDEMET, O duction of a atoxin on wheat, corn, rice and peanut. Journal of Veterinary Science, 11: 135–140. ¨ ., OGÆUZ, H., INAL , F. & NIZAMLIOGÆLU , F. (1996) Single DEMET, O and combination effects of dietary a atoxin and adsorbent Myco x Ò Plus (based PVPP active matter) on some performance parameters of broiler chickens. Journal of Veterinary Science, 12: 53–58. DONALDSON, W.E., TUNG , H.T. & HAMILTON, P.B. (1972) Depression of fatty acid synthesis in chick (Gallus domesticus) liver by a atoxin. Comparative Biochemistry and Physiology, 41: 843– 847. EDDS, G.T. & BORTELL, R.A. (1983) Biological effects of a atoxins: Poultry a atoxin and Aspergillus avus in corn, in: DIENER, U.L., A SQUIT, R.L. & DICKENS, J.W. (Eds) Bulletin of The

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