饲 粮 中 含 水 合 钠 钙 矽 酸 铝

对 抗 土 番 鸭 黄 麴 毒 素 症 之 影 响

郑永祥⁽¹⁾     林育安⁽¹⁾    李德南⁽¹⁾     吴辅佑⁽¹⁾

收件日期：83年10月7日；接受日期：83年11月24日

(1)国立宜兰农工专科学校畜产科。

摘   要：

        水合钠钙矽酸铝(HSCAS)为混合饲料使用之抗结块剂，添加於土番鸭饲粮中，以评估其对土番鸭黄麴毒素症之改善效果。试验使用400只土番鸭分成五处理组，黄麴毒素(AFB₁) 0 ppb＋0％ HSCAS (对照组)，AFB₁ 0 ppb＋2％ HSCAS，AFB₁ 400 ppb＋0％HSCAS，AFB₁ 400 ppb＋0.5％ HSCAS，AFB₁ 400 ppb＋2％ HSCAS，采高床栏饲，试验为期21天。试验饲粮粗蛋白质为18.8％，代谢能为2880 kcal／kg，试验期间饲料及饮水均自由采食。鸭只每周秤重一次，试验结束时采血并测定心、肝、脾、胰脏重量。

        AFB₁ 400ppb＋0% HSCAS组在只日采食量、饲料利用效率及平均每日增重显着较其他各处理组为差(P＜0.05，但含AFB₁ 0ppb＋2％ HSCAS或AFB₁ 400ppb＋2％ HSCAS与对照组比较时，无显着差异。血中GPT及GOT活性，以AFB₁ 400 ppb＋0％HSCAS显着高於AFB₁ 0 ppb＋ 2％ HSCAS及对照组。且AFB₁ 400ppb＋0.5％ HSCAS、AFB₁ 400ppb＋ 2％ HSCAS与对照组间亦无显着差异。相对脏器重量方面，AFB₁ 400ppb＋0％ HSCAS之心、肝、脾、胰脏均呈显着(P＜0.05)较AFB₁ 0ppb＋2％ HSCAS、对照组及AFB₁ 400ppb＋2％ HSCAS为重，但AFB₁ 400 ppb＋0.5％HSCAS於心、肝、脾脏重量仍较2％ HSCAS和对照组为重。黄麴毒素B₁组织残留方面，在胸肌中各处理组均量少无法检出，肝脏中仅AFB₁ 400ppb＋0％ HSCAS组及AFB₁ 400 ppb＋0.5％ HSCAS组可测得微量黄麴毒素B₁的残留。由结果发现饲料中0.5％ HSCAS可以降低400ppb AFB₁对土番鸭的毒害，而添加2％ HSCAS可以改善至与对照组无差异。HSCAS不仅可以作为饲料抗结块剂，亦可作为土番鸭黄麴毒素中毒预防管理之捷径(关键语：水合钠钙矽酸铝、黄麴毒素症、土番鸭)。

绪   言

        黄麴毒素(Aflatoxins)是一群极毒的化学物质，由黄麴菌属中Aspergillus flavus和Aspergillus parasiticus所产生的二次性代谢产物，其中以黄麴毒素B₁最为普遍且毒性最强。黄麴菌广泛存在於自然界中，易污染谷物导致动物慢性暴露於黄麴毒素中。黄麴毒素对家禽所造成之毒害，间接影响家禽生产之利润(Jones et al., 1982；Hamilton, 1984)。鸭只对黄麴毒素之感受性甚高，以雏鸭之感受性最强，故早期曾以雏鸭做为黄麴毒素之生物检定(bioassay)(Asplin and Carnaghan, 1961)。

        当饲料中污染黄麴毒素时，去毒的方法颇多；但至今仍无一可以达到大量处理且经济及实际之可行方法(Doyle et al., 1982；Norred, 1982)，近来使用一种非营养性吸附剂，水合钠钙矽酸铝(hydrated sodium calcium aluminosilicate, HSCAS)来控制或和黄麴毒素污染之饲料对家畜禽所造成的为害。此吸附剂应用在鸡(Scheideler, 1993)、猪(Harvey et al., 1989)、羊(Harvey et al., 1991)、火鸡(Kubena et al., 1991)等均可改善黄麴毒素之毒害，此种选择性之化学吸附剂，可在肠道内与黄麴毒素紧密结合形成稳定之HSCAS／Aflatoxin复合物，从而降低黄麴毒素之生物可利用性(Phillips et al., 1988)。Kubena et al. (1990a；1991)指出HSCAS可降低AFB₁对白肉鸡及火鸡雏之生长抑制作用与死亡率，据估计每毫克HSCAS可与483nm的AFB₁结合。土番鸭是本省最重要之肉用鸭种，迩来消费者之品质意识普遍抬头，对肉品之安全卫生相当敏感。故本试验即探讨土番鸭喂饲黄麴毒素时，饲粮中HSCAS可否提供诸如对其他家畜禽之保护效果及其对组织中黄麴毒素残留量的影响。

材料与方法

一、试验动物与及样品处理

        试验土番鸭雏为购自商业性孵化场之一日龄雏鸭，选择体重近且活力佳者，逢机分配至五处理组：黄麴毒素0 ppb＋0％ HSCAS (对照组)；黄麴毒素0 ppb＋2％ HSCAS ；黄麴毒素400 ppb＋0％ HSCAS；黄麴毒素400 ppb＋0.5％ HSCAS；黄麴毒素400 ppb＋2％ HSCAS，每组四重复，每重复20只，公母各半 。试验鸭只饲於网状高床，每栏放置一 250 W红外线灯泡予以保温。试验期0～3周龄，试验期间饲料和饮水均自由采食。试验饲粮含蛋白质18.8％；代谢能2880 kcal／kg (表1)。黄麴毒素与HSCAS均直接外加於试验饲粮。

Table  1.   Composition    of     basal    diet.

大  豆  粉 (44％)

鱼        粉 (60％)

麸        皮

磷酸氢钙

石灰石粉

大  豆  油

食　　盐

L-离胺酸

DL-甲硫胺酸

氯化胆硷，50％

维生素预混物^A

矿物质预混物^A

合        计

Soybean meal, 44%

Fish meal, 60%

Wheat branl

Dicalcium phosphate

Limestone, pulverized

Soybean oil

Salt

L-Lysine

DL-methionine

Choline chloride, 50%

Vitamin premix ^A

Mineral premix ^A

Total

24.50

3.00

5.00

1.30

0.75

0.60

0.30

0.10

0..07

0.08

0.30

0.20

100.00

粗蛋白质，％

代谢能，kcal／kg

含硫胺基酸，％

钙，％

有效磷，％

Crude protein, %

ME, kcal/kg

Sulfur amino acid, %

Calcium, %

Available phosphorus, %

18.8

2880

0.73

0.86

0.50

        试验期间每周秤重一次，试验结束时采血，经远心分离後血清供血液生化学分析，血清经血液生化分析仪(Hitachi, 705)分析血清中麸胺草醋酸转胺鶤(Glutamate oxaloacetate transaminase；GOT)、麸胺丙酮酸转胺鶤(Glutamate pyruvate transaminase；GPT)、硷性磷酸鶤(Alkaline phosphatase；ALP)之活性及总胆红质(Total bilirubin)浓度，鸭只经屠宰、剖腹，记录心脏、肝脏、脾脏、胰脏之重量。另取右叶肝脏一小块及右胸肌供组织残留量分析。

二、黄麴毒素产制及组织残留量分析

        黄麴毒素粗制品之产制系综合Shotwell et al. (1966)；Smith and Hamilton (1969)及West et al. (1973)之方法。粗制品经磨碎、混匀、采样後以AOAC (1984)方法萃取及清净化後，再以性能液相层析法(HPLC)定性定量之。粗制品中所含黄麴毒素B₁做为饲粮添加之依据。

        肝脏及胸肌中之黄麴毒素B₁残留量，采用Tarter et al. (1984)之检验法萃取，再以HPLC定量之。

三、统计分析

        所有试验资料使用一般线性模式(General linear model procedure)行变方分析後，再以邓肯氏新多项变域法(Duncan new multiple range test)比较各组间之差异显着性。

结果与讨论

        水合钠钙矽酸铝(HSCAS)为白陶土的钠钙化合物，在美国食品药物管理局登记为饲料抗结块剂(anticaking agent)，近年来使用普遍。SHCAS在肠道中选择性吸附黄麴毒素B₁ (AFB₁)，减少黄麴毒素为动物体所吸收。HSCAS之吸附作用仅对黄麴毒素呈特异性吸附，对T-2 toxin (Kubena et al., 1990a)，赭麴毒素(ochratoxin)(Huff et al., 1992)，Diacet oxyscirpenol (Kubena et al., 1993)等均无吸附效果。

        饲料中含HSCAS与黄麴毒素对0～3周龄土番鸭生长性能之影响列於表2。试验期间鸭只除AFB₁ 0 ppb＋2％HSCAS组及AFB₁ 400 ppb＋0.5％ HSCAS组因逃出冻死外，其馀均无死亡。在只日采食量及全期平均每日增重方面，AFB₁ 400 ppb＋0％ HSCAS组均呈显着(P＜0.05)较其他各处理组为差。AFB₁ 400 ppb＋2％ HSCAS组只日采食量、全期平均每日增重及饲料利用效率与对照组间无显着差异，AFB₁ 400ppb＋0.5％ HSCAS组除全期平均日增重较对照组为差(P＜0.05)之外，饲料利用效率与对照组间并无显着效果。平均日增重随着周龄增加，AFB₁ 400 ppb＋2％ HSCAS组有改善的效果。AFB₁ 0 ppb＋2％ HSCAS组生长性能影响与对照组间均无显着差异。据Harvey et al. (1989)之报告指出日粮中添加0.5％ HSCAS或2％ HSCAS均可改善饲料中AFB₁ (3ppm)所造成猪只的毒害，但以2％ HSCAS效果较佳。此与本试验之结果一致。Kubena et al. (1990a)以肉鸡(0～3)之实验发现0.5％ HSCAS可以改善7.5 ppm AFB₁所导致只日增重和饲料利用率变差的情形，但额外添加0.5％活性炭(charcoal)却无保护效果。HSCAS之保护机制可能为黄麴毒素B或G系列(AFB₁，B₂，B_2a，G₁，G₂，G_2a)含有β-内脂环(β-ketolactone)或α-双内酯环(α-bislactone)有较佳之吸附性，而且黄麴毒素之β-carbonyl系统与HSCAS边位(edge site)之金属离子形成稳定化合物(Sarr et al., 1990；1991)所致。

Table 2. Effect of HSCAS on the growth performance of mule ducklings fed diets containing aflatoxin from 0 to 3 weeks of age.

Items   HSCAS,  ％

SE

Daily feed intake (g／bird／day)

平均每日增重

(Avg. daily gain (g／bird／day)

第一周

0-1 wks

第二周

1-2 wks

第三周

2-3 wks

全    期

0-3 wks

饲料利用效率

Food conversion (Feed/ gain)

19.09^a

38.40^a

49.78^a

35.76^a

1.66^b

18.65^a

37.35^a

49.88^a

35.29^a

1.67^b

11.08^d

13.36^c

28.74^c

17.73^c

2.33^a

13.41^c

28.60^b

39.68^b

27.23^b

2.10^ab

16.43^b

37.67^a

49.12^a

34.40^a

1.70^b

1.84

4.32

9.86

3.44

0.12

Means in the same row with different superscripts differ significantly (P＜0.05).

Table 3.   Effect    of    HSCAS    on     the    relative    organ     weight    and    aflatoxin     tissue    residue    of     mule    ducklings    fed     diets    containing    aflatoxin     from    0    to    3     weeds     of     age.

Items HSCAS, %

SE×10^-4

Heart weight／Body weight

肝脏／体重(×10^-2)

Liver weight／Body weight

脾脏／体重(×10^-3)

Spleen weight／Body weight

胰脏／体重(×10^-3)

Pancreas weight／Body weight

2.89^b

1.05^c

5.41^b

2.95^b

1.21^c

5.54^b

4.27^a

3.74^a

7.23^a

4.10^a

2.34^b

6.00^b

3.58^ab

1.43^c

5.80^b

0.92

6.81

0.13

Aflatoxin residue in lever, ppb

胸肌之B₁残留量

Aflatoxin residue in breast muscle, ppb

0

0

0

0

0

－

a.b.c.：同行内数值标有不同字母者，表示差异显着(P＜0.05)。

Means in the same row with different superscripts differ significantly (P＜0.05).

        黄麴毒素作用之标的器官为肝脏。Bryden and Cumming (1980)试验指出鸡只黄麴毒素中毒时，肝脏有不同程度之脂肪浸润(fatty infiltration)及肝细胞退化。Merkley et al. (1987)与Huff et al. (1986)均指称初期黄麴毒素症因肝细胞退化使肝重降低，当持喂与黄麴毒素时肝脏明显肿大，肝重、体重比显着提高。

        在组织残留量方面，肝脏中AFB₁残留以AFB₁ 400ppb＋0％ HSCAS组最高(0.021ppb)；AFB₁ 400 ppb＋0.5％ HSCAS组亦有少量残留。而AFB₁ 400 ppb＋2％HSCAS组则无残留，显示2％ HSCAS可充分吸附AFB₁。由此一结果推测可能与HSCAS和AFB₁之间机会性接触有关。在胸肌残留量，各处理组均无AFB₁检出。以白肉鸡(沈等，1987)、土番鸭(沈等，1988)与有色肉鸡(吴等，1990)之试验发现胸肌中无残留，仅肝脏少量残留，此外；吴等也指出生长猪(吴等，1988)及肥育猪(吴等，1989)喂饲黄麴毒素仅肝脏中残留少量，肌肉亦未检出。以上报告与本试验之结果相吻合。

        饲粮中含HSCAS与黄麴毒素对0～3周龄土番鸭血液生化值之影响列於表4。在总胆红质及硷性磷酸鶤各处理组间无显着差异，此一结果与Kubena et al. (1990b)之结果一致。在麸胺草醋酸转胺鶤(GOT)及麸胺丙酮酸转胺鶤(GPT)活性方面，AFB₁ 400 ppb＋0％ HSCAS组之GOT活性均高於其他处理组，而0.5％或2％ HSCAS添加均可见改善之效果。在GPT活性方面，AFB₁ 400 ppb＋0％ HSCAS组之GPT活性均显着高於其他处理组(P＜0.05)，而0.5％或2％ HSCAS添加亦有改善之效果。Manning et al. (1990)指出黄麴毒素使得鸡只血中GPT及GOT活性升高。由於GPT为存於肝细胞中之特殊转胺鶤；肝细胞受损时，细胞膜通透性增加使血中GPT活性随之升高，而黄麴毒素对肝之损害具特异性，故以GPT较为敏感。Harvey et al. (1989)试验报告指出黄麴毒素3ppm喂与生长猪四周，GOT、GPT活性显着升高，而日粮中添加0.5％或2％ HSCAS时GPT活性下降。

表四   饲   粮    中   含   水    合   钠   钙   矽   酸    铝   与   黄   麴   毒    素   对   0   ～   3    周   龄   土   番   鸭    血   液   生   化   值    之   影   响

Table 4. Effect of HSCAS on the blood biochemical index of mule ducklings fed diets containing aflatoxin from 0 to 3 weeks of age.

Items   HSCAS,  ％

SE

Total bilirubin

麸胺草醋酸转胺鶤

GOT* (U／L)

麸胺丙酮酸转胺鶤

GPT* (U／L)

硷性磷酸

鶤ALP* (U／L)

46.10^b

39.10^b

435.20

47.52^b

40.47^b

412.32

97.44^a

63.17^a

461.67

62.72^ab

41.11^b

450.55

67.41^ab

42.94^b

439.70

9.95

6.61

6.67

^a.b.：同行内数值标有不同字母者，表示差异显着(P＜0.05)。

        Means in the same row with different superscripts differ significantly (P＜0.05).

        综合本试验之结果显示，饲粮中0.5％或2％ HSCAS可以降低黄麴毒素400 ppb 对土番鸭之危害，但以2％ HSCAS 添加量最佳，且此一添加量对土番鸭之生长性能并无不良之影响。Chung et al. (1990)试验指出HSCAS不影响锰、维生素A及B2之吸收，但会降低锌的利用率。Chung and Baker (1990)也指出0.5％或2％ HSCAS对植酸磷或无机磷之利用并不影响。是否对其他营养分之吸收有负面影响仍待探讨。

志   谢

        本试验承行政院国家科学委员会经费补助(HSC83-0117-C-197-007-B)，谨此致谢。

参考文献

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Efficacy   of    Hydrated   Sodium   Calcium   Aluminosilicate    Against   the   Aflatoxicosis   of    Mule   Ducklings

Yeong-Hsiang Cheng⁽¹⁾, Yu-An Lin⁽¹⁾, Der-Nan Lee⁽¹⁾ and Fu-Yu. Wu⁽¹⁾

Received Oct. 7. 1994; Accepted Nov. 24. 1994

ABSTRACT

        The objective of this study was to investigate the use of hydrated sodium calcium aluminosilicate (HSCAS) to minimize the detrimental effects of aflatoxin B1 (AFB₁) on mule duckling, based on their growth performance, relative visceral organ weight, blood biochemical index and aflatoxin residue in tissues.

     Four hundred duckling were randomly divided into five groups which were fed diets containing AFB₁ 0 ppb ＋0％ HSCAS (control), AFB₁ 0 ppb ＋2％ HSCAS, AFB₁ 400 ppb＋0％ HSCAS, AFB₁ 400ppb ＋0.5％ HSCAS, or AFB₁ 0 ppb ＋2％ HSCAS.

    The results showed that 400 ppb AFB₁ decreased growth performance, increased the activities of GOT and GPT in serum, and increased the relative visceral organ weight (P＜0.05). The addition of 2％ HSCAS could overcome these adverse effects completely, but 0.5％ HSCAS could not.

        Trace amounts of AFB₁ were detected in the liver of mule ducklings fed diets containing AFB₁ 400 ppb and AFB₁ 400 ppb ＋ 0.5％ HSCAS. No AFB₁ residues were detected in the breast muscle of all treatments. These findings suggested that 2% HSCAS could modulate the toxicity of aflatoxin in mule ducklings in animals. (Key Words: Hydrated sodium calcium aluminosilicate, Aflatoxicosis, Mule Ducklings)

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⁽¹⁾ Department of Animal Science, National I-Lan Institute of Agriculture and Technology.