200423950 玖、發明說明： 【發明所屬之技術領域】 本發明係關於包含鳥類所產蛋或其處理物與活菌劑之抗 腹瀉症組合物、包含該組合物之抗腹瀉症醫藥組合物、食 品及飼料，以及藉由於家畜投藥鳥類所產蛋或其處理物與 活菌劑而預防及治療腹瀉症之方法。 【先前技術】 腹离症為人類及家畜常見疾病之一，但於幼小體弱個 體、特別於哺乳期中引發脫水症或其他合併症、導致嚴重 狀態之情形多有發生。腹瀉症之原因可分為病毒或細菌引 起之感染性及環境、壓力、食物等引起之非感染性兩類。 特別地感染性腹瀉症向其他個體傳播，常常引起群體中大 多數個體受害。 作為感染性腹瀉症之主要病原體，可例舉病毒與細菌。 該等病原體隨食物或飲料經口侵入腸道、附著於腸黏膜繁 殖，引起發病。又，2種以上病原體復合感染導致症狀更為 嚴重化之情形並非罕見。 作為病毒性腹瀉症之病原體，已知輪狀病毒、小病毒、 '片、病毋"右沃烏病骨、杯狀病毒、腺病毒、星狀病毒等。 作為細菌性腹瀉症之病原體，已知痢疾志贺桿菌、沙氏桿 原性A腸桿菌、毒素原性大腸桿菌'霍亂孤菌、: 炎弧菌、耶氏桿菌、葡萄球菌、金黃色葡萄球菌、綠膿桿 菌’及屬於彎曲g屬、螺旋桿菌屬及芽胞梭菌屬之細菌等。 由上述病原體引發之各種腹瀉症為引起各種動物劇烈唱 90052.doc ^00423950 吐及腹瀉之疾病。特 打別疋％炎型，確認為初期嘔吐、 之内轉為腹;寫、繼而時當领 吋㊆便血，伴隨食欲不振、無精打姿 急速脫水、體溫上升等_ 本、 寺f月形。針對该等腹瀉症中之一邱八 現在疫苗正在普及。 ϋ刀’ …、而與以別相比，該腹寫症疫苗之防治效果存在問題 對於防治腸道感染症重要 吨。 压垔要的疋如何產生IgA抗體。現在正 吏用之m疫田知用肌肉注射使igG抗體能夠產生， 於腸道内IgA抗體之產生量少，得不到理想效果。金：相 應：活菌疫苗經Π投藥，IgA抗體之產生量多，防治效果得 到可&現在主要針對病毒性腹蕩症使用活菌疫苗，但 是其效果亦不能說十分理想。另，料、細菌，於病原性復 原問題方面被認可之活芦钴彡|、 J又居&田很少。另一方面，抗生素亦被 作為腹m治療藥使用。然而對於細菌性腹瀉症，存在 著由於使用抗生素而產生雜g之問題。再者，為促進發 育及預防腹萬’特別於家畜飼料中添加抗生素之做法，於 環境污染方面也存在著問題。 、 另一方面，已知雞蛋抗體與疫苗不同，為適於經口投藥 之抗體。作為使用雞蛋抗體之醫藥組合物，已公開藉由以 引發豬大腸桿菌症之腸道毒素原性大腸桿菌之987P、K88 及Κ99抗原、及引發牛大腸桿菌症之腸道毒素原个生大腸桿菌 之Κ99抗原中任一種以上之抗原對雞進行免疫、從該免疫雞 所產蛋之至少含有蛋黃之部分中回收抗體而得到之對上述 抗原具有特效、包含多株抗體之大腸桿菌症之經口預防劑 及治療劑（特許第2034005號）。又，亦公開包含預先接種食 90052.doc 200423950 、蛋黃或含有抗體 之抗體之抑制食物 物中毒菌之雞獲得免疫後所產蛋之全蛋 部分、其中含有對該食物中毒菌有特效 中毒菌材料等（特許第2615673號）。 又，已公開包含從狗小病毒免疫過之雞之蛋黃所得抗體 之抗小病毒感染症組合物作為治療狗小病毒感染症之：藥 組合物⑽開平8_259462)。又，已公開包含從豬流行性腹= 病毒免疫過之牛採集之乳或其乳成分、及從該病毒免疫過 之雞採集之雞蛋蛋黃或雞蛋抗體之醫藥組合物，作為治療 豬机行性腹瀉病毒感染症之醫藥組合物（特開平 10-265393)。又，已公開丹寧類與對感染性微生物或該等微 生物產生之毒素有特效之抗體併用，作為對家畜、家禽及 寵物之非感染性腹瀉症及感染性腹瀉症有效之飼料組合物 (特開平 7-067544)。 另一方面，有报多文獻有關活菌劑之功效，亦有報導活 菌劑對治療動物單純性腹瀉有效。又，已公開包含以腸導 出血性大腸桿菌之菌體或類毒素化佛羅毒素（verotoxin)作 為抗原對雞免疫、從該雞所產蛋所得之抗體與1種以上活菌 劑併用，作為有效成分之腸導出血性大腸桿菌感染症之預 防劑及治療劑（特開平1〇_298105)。 但是，尚未公開於細菌及病毒引發之各種腹瀉症發揮優 /、預防效果之同日守，可減輕腹〉寫症特有之腹填、喂吐及體 重減輕等症狀、具有優異治療效果之組合物或方法。 【發明内容】 本發明之課題為提供一種於預防腹瀉症發病及改善腹瀉 90052.doc 200423950 症特有之腹瀉、嘔吐及體重減輕之症狀同時，無耐性菌發 生之問題，有效預防及治療腹瀉症之組合物。 本發明者們為解決上述課題銳意研究發現’藉由於動物 投藥包含以腹瀉症病原體為抗原對鳥類進行免疫所得蛋戋 其處理物與活菌劑之組合物，可解決上述課題，從而完: 本發明。通常認為以腹瀉症病原體為抗原對鳥類進行免疫 所得蛋或其處理物中所含之抗體，藉由附著於病原體、： 病原體中和而使其病原性消《；或者藉由附著於病原體細 胞黏著素而阻止病原體附著於腸導細胞而使其病原性消 失。再者’藉由與活菌劑合併投藥，腸内菌叢得到改善同 時，上述抗體更加有效作用’發揮優異之預防及治療腹瀉 錢果。X，藉由活菌劑與本發明之抗腹瀉症抗體混合投 樂，投藥少量抗體也可發揮優異之乘數效應效果。 >本毛明係關於包含腹瀉症病原體或來自腹瀉症病原體之 抗原免疫過之鳥類所產蛋或其處理物與活菌劑之抗腹濡症 組合物。 原蟲所組成之群中所選之如上所述之組合物„ 又，本發明係關於其中活菌劑為乳酸菌之如上任一項 述之組合物。 又，本發明係關於包含 3如上任一項所述之組合物之抗‘ 渴症醫療組合物。 又，本發明係關於包 S如上任一項所述之組合物之飼料 又，本發明係關於包 ★上任一項所述之組合物之食品 90052.doc 200423950 又，本發明係關於藉由於動物投藥包含腹瀉症病原體或 來自腹瀉症病原體之抗原免疫過之鳥類所產蛋或其處理物 與活菌劑而預防及治療腹瀉症之方法。 又’本發明係關於其中動物為人以外之動物之如上所述 之治療方法。 本發明中，作為以腹瀉症病原體或來自腹瀉症病原體之 抗原進行免疫之鳥類，沒有特別地限定，例如雞、鶉等。 從抗體之產量方面考慮，優選利用雞、特別為產蛋雞。 本發明中術語腹瀉症具有本技術領域中常用之意義，即 指異常地反復從腸導排泄液狀或半固體糞便之症狀。本發 明中’腹瀉症病原體係成為引發動物腹瀉症原因之細菌、 真菌、原蟲及病毒等微生物。腹瀉症病原體，並不限於該 等舉例，包含輪狀病毒、裏奥病毒、小病毒、冠狀病毒、 腸病毒、諾沃克病毒、杯狀病毒、腺病毒、星狀病毒、黑 頭病毒及流行性感冒病毒等病毒；屬於志贺桿菌屬（病疾志 贺桿菌等）、沙氏桿菌屬（Salmonella dublin、Salmonella typhimurium等）、大腸桿菌屬（病原性大腸桿菌、病毒原性 大腸桿菌等）、弧菌屬（霍亂菌、腸炎弧菌等）、耶氏桿菌屬 (Yersinia enterocolitica等）、葡萄球菌屬（金黃色葡萄球菌 等）、假單胞菌屬（綠膿桿菌等）、彎曲菌屬（Campylobacter fetus等）、螺旋桿菌屬、芽胞梭菌屬（Clostridium perfringens 等）、及螺旋體屬（Serpulina hyodysenteriae等）、產氣單胞菌 屬（Aeromonas hydrophila 等）、鄰單胞菌屬（Plesiomonas shigelloides等）及芽孢桿菌屬（Bacillus cereus等）之細菌；屬 90052.doc -10· 200423950 於梨形鞭毛蟲(Giardia)屬、艾美球蟲屬（Eimeriascabra等）、隱胞子 蟲（Cryptosporidium)屬、弓蟲（Toxoplasma)屬、滴蟲（Trichomonas) 屬、五鞭毛蟲(Pentatrichomonas)屬、聯同形球蟲(Isospora)屬、内 阿米巴（Entamoeba)屬、及纖毛蟲屬（Balantidium)、球蟲（coccidium) 類等之原蟲等。 本發明中來自腹瀉症病原體之抗原係由上述腹瀉症病原 體之部分構成要素組成之肽，包含例如腹瀉症病原體之表 面抗原、外膜蛋白質、黏著素、侵入素、毒素、代謝產物 及病原因子（etiologic agent)，也包含由該等中之一部分所 組成之物。本發明中，優選使用該等來自腹瀉症病原體之 抗原。腹濕症病原體及由其而來之抗原包含於細胞溶解物 中，其精製物及未精製物均可使用，優選使用精製物。抗 原之精製方法，可單獨利用硫銨分化法、柱式層析法、電 泳法等通常方法，亦可單獨或2種以上組合使用。又，柱式 層析法中可使用1種層析柱，亦可2種以上層析柱組合使 用。通常，柱式層析法中可利用離子交換層析法、膠濾層 析法、逆相層析法、親和層析法等。 更為具體而言，利用本發明之組合物可預防及治療之腹 瀉症及成為引發該腹瀉症之原因之病原體，並不限於如下 例舉之實例。對於牛，有病毒性腹瀉黏膜病毒（BVDV)引發 之牛病毒性腹瀉黏膜病、牛冠狀病毒引發之牛冠狀病毒 病、牛輪狀病毒引發之牛輪狀病毒病、牛小病毒引發之牛 小病毒病、牛腸病毒引發之牛腸病毒病、Clostridium perfringens引發之牛腸導血毒症、Campylobacter fetus引發 90052.doc -11 - 200423950 之牛彎曲菌病、Salmonella dublin、S. typhimurium、S. enteritidis等引發之牛沙氏桿菌症、毒素原性大腸桿菌引發 之子牛大腸桿菌性腹瀉、Cryptosporidium parvum引發之牛 隱胞子蟲症等。對於綿羊，有Clostridium perfringens引發 之綿羊赤痢及綿羊芽胞梭菌症等。對於馬，有馬輪狀病毒 引發之馬輪狀病毒病等。對於豬，有傳染性胃腸炎病毒引 發之傳染性胃腸炎、豬流行性腹瀉病毒引發之豬流行性腹 瀉、裏奥病毒引發之裏奥病毒病、大腸桿菌引發之豬大腸 桿菌症、Salmonella choleraesuis、S. typhimurium、S· typhisuis、 S. derby等引發之沙氏桿菌症、Serpulina hyodysenteriae 引 發之豬赤痛j、Clostridium perfringens引發之豬腸導血毒 症、Yersinia enterocolitica、 Y. pseudotuberculosis引發之 豬耶氏桿菌症、Campylobacter mucosalis、C. hyointestinalis 引發之腸腺腫症侯群、Eimeria scabra、Ε· debliecki引發之 豬球蟲病等。對於狗及貓，有狗小病毒引發之狗小病毒病、 狗艰狀病毒引發之狗趙狀病毒病、f苗泛白血球減少症病毒 (小病毒）引發之I苗泛白血球減少症、I苗白血病病毒引發之猶 白金病、Campylobacter jejuni、C· coli引發之彎曲菌症、 Isospora canis、I. ohioensis、I. felis、I. rivolta、Sarcocystis、 Eimeria、Toxoplasma 引發之狗描球蟲病、Toxoplasma gondii 引發之狗描弓蟲病、Giardia canis、G. cati引發之狗f苗梨形 鞭毛蟲症、Pentatrichomonas hominis引發之狗猶滴蟲病、 Cryptosporidium parvum引發之狗 f苗隱胞子蟲症、Entamoeba histolytica引發之狗貌阿米巴症、Balantidium coli引發之狗 90052.doc -12- 200423950 纖毛蟲症等。對於鳥類，有Duck virus enteritis virus引發之 家鴨病毒性腸炎、火雞冠狀病毒引發之火雞冠狀病毒腸 炎、輪狀病毒引發之鳥輪狀病毒病、出血性腸炎病毒引發200423950 发明 Description of the invention: [Technical field to which the invention belongs] The present invention relates to an anti-diarrheal composition containing an egg produced by a bird or a processed product thereof and a live bacterial agent, a pharmaceutical composition against diarrhea containing the composition, and food And feed, and methods for preventing and treating diarrhea by administering eggs produced by birds or their treatments and live bacteria to livestock. [Prior art] Abdominal diarrhea is one of the common diseases of humans and livestock, but it often causes dehydration or other comorbidities in young and fragile individuals, especially during breastfeeding. The causes of diarrhea can be divided into two types: infectivity caused by viruses or bacteria, and non-infectious causes caused by the environment, stress, and food. In particular, infectious diarrhea is transmitted to other individuals and often causes the majority of individuals in the group to suffer. The main pathogens of infectious diarrhea include viruses and bacteria. These pathogens invade the intestinal tract with food or drink, attach to the intestinal mucosa, and cause disease. In addition, it is not uncommon for symptoms to worsen with a combination of two or more pathogens. As pathogens of viral diarrhea, rotaviruses, parvoviruses, "tablets, disease-free" bones, caliciviruses, adenoviruses, astroviruses, and the like are known. As the causative agent of bacterial diarrhea, Shigella dysenteriae, Enterobacter aeruginosa serovar, toxinogenic E.coli's cholera, Vibrio spp, Yersinia, Staphylococcus aureus , Pseudomonas aeruginosa 'and bacteria belonging to Genus Campylobacter, Helicobacter and Clostridium. The various diarrhea caused by the above pathogens are diseases that cause various animals to sing violently. 90052.doc ^ 00423950 Vomiting and diarrhea. Specially do not 疋% inflammation type, confirmed as initial vomiting, turned into the abdomen; write, and then when the collar ㊆ ㊆ blood in the stool, accompanied by loss of appetite, impotence, rapid dehydration, temperature rise, etc. Qiu Ba, one of these diarrhea, is now spreading vaccines. ϋ 刀 '... Compared with others, there is a problem with the efficacy of the abdominal write-off vaccine for the prevention and treatment of intestinal infections. It is important to know how to produce IgA antibodies. Nowadays, it is known that igG antibody can be produced by intramuscular injection, and the amount of IgA antibody produced in the intestine is small, and the ideal effect is not obtained. Gold: Correspondence: The live bacteria vaccine is administered via Π, and the production of IgA antibodies is large, and the prevention and treatment effect is acceptable. Currently, the live bacteria vaccine is mainly used for viral diarrhea, but its effect cannot be said to be very satisfactory. In addition, there are few live reeds and bacteria that are recognized for pathogenic restoration problems, bacteria, and bacteria. On the other hand, antibiotics are also used as a therapeutic agent for abdominal m. However, in the case of bacterial diarrhea, there is a problem that g is caused by the use of antibiotics. Furthermore, in order to promote the development and prevention of abdominal diseases, especially the practice of adding antibiotics to livestock feed, there are also problems with environmental pollution. On the other hand, it is known that egg antibodies are different from vaccines and are suitable for oral administration. As a pharmaceutical composition using egg antibodies, 987P, K88, and K99 antigens of enterotoxin-producing Escherichia coli causing porcine colisis, and enterotoxin-producing Escherichia coli causing bovine colisis have been disclosed. Oral administration of any one or more of the K99 antigens to immunize chickens, recover antibodies from at least the yolk-containing portion of eggs produced by the immunized chickens, and obtain oral coliforms that have specific effects on the above antigens and contain multiple antibodies Prophylactic and therapeutic agents (Patent No. 2034005). In addition, it also discloses a whole egg portion containing eggs inoculated with chickens that pre-vaccinated 90052.doc 200423950, egg yolk, or antibody-containing antibodies to suppress toxic bacteria in food, and contains materials that are particularly effective against toxic bacteria in food Etc. (Charter No. 2615673). Also, an anti-parvovirus infection composition containing an antibody obtained from the egg yolk of a chicken immunized with canine parvovirus has been disclosed as a medicinal composition for treating canine parvovirus infection: kaikaihei 8_259462). In addition, a pharmaceutical composition containing milk collected from porcine epidemic = virus-immunized cows or milk components thereof, and egg yolk or egg antibodies collected from chickens immunized with the virus has been disclosed as a method for treating porcine autonomy Pharmaceutical composition for diarrhea virus infection (Japanese Patent Application Laid-Open No. 10-265393). In addition, it has been disclosed that tannins are used in combination with antibodies having specific effects on infectious microorganisms or toxins produced by these microorganisms as feed compositions effective for non-infectious diarrhea and infectious diarrhea in livestock, poultry, and pets (specifically, Kaiping 7-067544). On the other hand, there have been many reports on the efficacy of live bactericides, and there have been reports of effective bacterial agents for treating simple diarrhea in animals. In addition, it has been disclosed that the use of intestinal blood-derived Escherichia coli cells or toxin-like verotoxin as an antigen to immunize chickens, and the use of antibodies obtained from eggs produced by the chickens in combination with one or more live bacteria agents is effective. The intestine of the component leads to a preventive agent and a therapeutic agent for bloody E. coli infection (Japanese Patent Laid-Open No. 10-298105). However, it has not yet been disclosed that diarrhea caused by bacteria and viruses exhibits excellent / preventive effects on the same day, and can reduce symptoms such as abdominal filling, feeding and vomiting that are unique to dysentery, and a composition having excellent therapeutic effect or method. [Summary of the Invention] The object of the present invention is to provide a method for effectively preventing and treating diarrhea while preventing the onset of diarrhea and improving the symptoms of diarrhea 90052.doc 200423950 which are characteristic of diarrhea, vomiting, and weight loss. combination. In order to solve the above-mentioned problems, the inventors made an intensive study and found that 'the composition of a processed product and a viable agent of egg crust obtained by immunizing birds with diarrhea pathogens as antigens due to animal administration can solve the above problems, thereby completing: invention. It is generally believed that the antibodies contained in eggs or their processed products obtained by immunizing birds with diarrhea pathogens as antigens can be eliminated by attaching to the pathogens: neutralizing the pathogens; or by attaching cells to the pathogens It can prevent pathogens from attaching to intestinal cells and make them pathogenic. Furthermore, 'by injecting the drug in combination with a live bacteria agent, the intestinal flora is improved, and the above-mentioned antibodies are more effective.' It exerts excellent prevention and treatment of diarrhea. X. By mixing the live bacteria agent with the anti-diarrhea antibody of the present invention, even a small amount of the antibody can also exert an excellent multiplier effect. > The present mollium is an anti-diarrheal composition comprising an egg produced by a bird immunized with an diarrhea pathogen or an antigen derived from the diarrhea pathogen, or a processed product thereof, and a live bacteria agent. The composition as described above selected from the group consisting of the protozoa. Furthermore, the present invention relates to the composition according to any one of the above, wherein the living bacteria agent is a lactic acid bacterium. Furthermore, the present invention relates to the composition comprising any one of the above 3 The anti-thirsty medical composition of the composition according to item 1. In addition, the present invention relates to a feed containing the composition according to any one of the preceding items, and the present invention relates to the composition according to any one of the above-mentioned items. Food 90052.doc 200423950 In addition, the present invention relates to a method for preventing and treating diarrhea by administering an egg produced by a bird immunized with a diarrhea pathogen or an antigen derived from a diarrhea pathogen, or a processed product thereof, and a viable bacterial agent. Also, the present invention relates to a treatment method as described above in which the animal is an animal other than human. In the present invention, the bird immunized with the diarrhea pathogen or an antigen derived from the diarrhea pathogen is not particularly limited, such as chicken , Quail, etc. In terms of the production of antibodies, chickens, especially laying hens are preferably used. The term diarrhea in the present invention has a meaning commonly used in the technical field, that is, Refers to the symptoms of abnormally repeated drainage of liquid or semi-solid feces from the intestine. In the present invention, the 'diarrheal pathogenic system becomes microorganisms such as bacteria, fungi, protozoa, and viruses that cause diarrhea in animals. The pathogen of diarrheal disease is not limited to Examples include viruses such as rotavirus, rio virus, parvovirus, coronavirus, enterovirus, Norwalk virus, calicivirus, adenovirus, astrovirus, blackhead virus, and influenza virus; belonging to Shiga Bacillus (Shigella spp., Etc.), Salmonella (Salmonella dublin, Salmonella typhimurium, etc.), Escherichia coli (pathogenic E. coli, virogenic E. coli, etc.), Vibrio (Cholera, Enterococcus Bacteria, etc.), Yersinia enterocolitica, etc., Staphylococcus (staphylococcus aureus, etc.), Pseudomonas (pseudomonas aeruginosa, etc.), Campylobacter (Campylobacter fetus, etc.), Helicobacter, Clostridium perfringens, etc., and Serpulina hyodysenteriae, etc., Aeromonas hydrophila, etc.), bacteria of the genus Plesiomonas shigelloides, etc. and Bacillus cereus, etc .; genus 90052.doc -10 · 200423950 in the genus Giardia, Eimeria ( Eimeriascabra, etc.), Cryptosporidium, Toxoplasma, Trichomonas, Pentarichomonas, Isospora, Entamoeba Protozoa of the genus, and the genus Balantidium, coccidium, etc. The antigen derived from the diarrhea pathogen in the present invention is a peptide composed of the above-mentioned constituent elements of the diarrhea pathogen, including, for example, a surface antigen of the diarrhea pathogen, an outer membrane protein, an adhesin, an invasin, a toxin, a metabolite, and a pathogen ( etiologic agent), also includes things that consist of one of these. In the present invention, such antigens derived from diarrhea pathogens are preferably used. The gastroenteritis pathogen and the antigen derived therefrom are contained in the cell lysate, and both the purified product and the unrefined product can be used, and the purified product is preferably used. The purification method of the antigen may be a general method such as an ammonium sulfate differentiation method, a column chromatography method, or an electrophoresis method, or it may be used alone or in combination of two or more. In the column chromatography, one type of column may be used, or two or more types of columns may be used in combination. Generally, column chromatography can use ion exchange chromatography, gel filtration chromatography, reverse phase chromatography, affinity chromatography, and the like. More specifically, the diarrhea that can be prevented and treated by the composition of the present invention and the pathogen that causes the diarrhea are not limited to the following examples. For cattle, there are bovine viral diarrheal mucosal diseases caused by viral diarrhea mucosal virus (BVDV), bovine coronavirus disease caused by bovine coronavirus, bovine rotavirus disease caused by bovine rotavirus, and bovine parvovirus caused by bovine parvovirus. Virus disease, bovine enterovirus disease caused by bovine enterovirus, bovine intestinal hemorrhagic disease caused by Clostridium perfringens, Campylobacter fetus causing 90052.doc -11-200423950 bovine Campylobacter disease, Salmonella dublin, S. typhimurium, S. enteritidis, etc Bovine salmonellosis, Bovine Escherichia diarrhea caused by toxinogenic E.coli, Cryptosporidium parvum, etc. For sheep, Clostridium perfringens causes Sheep smut and Clostridium sheep disease. For horses, there is rotavirus disease caused by rotavirus. For pigs, there are infectious gastroenteritis caused by infectious gastroenteritis virus, swine epidemic diarrhea caused by porcine epidemic diarrhea virus, Rio virus disease caused by Rio virus, swine coli disease caused by E. coli, Salmonella choleraesuis, S. typhimurium, S. typhisuis, S. derby, etc., S. typhiurium, S. typhisuis, S. derby etc. Bacillus, Campylobacter mucosalis, C. hyointestinalis-induced intestinal adenoma syndrome, Eimeria scabra, E. debliecki-induced swine coccidiosis, etc. For dogs and cats, there are canine parvovirus disease caused by canine parvovirus, canine poliovirus disease caused by dog hard virus, and pandemic leukopenia caused by pandemic leukopenia virus (parvovirus). Leukemia virus-induced Jewish platinum disease, Campylobacter jejuni, Campylobacter caused by C. coli, Isospora canis, I. ohioensis, I. felis, I. rivolta, Sarcocystis, Eimeria, Toxoplasma-induced dog schistosomiasis, Toxoplasma gondii Canine toxoplasmosis caused by dogs, Giardia canis, Canine f. Piriformis caused by G. cati, Canine trichomoniasis caused by Pentatrichomonas hominis, C. canis caused by Cryptosporidium parvum, Entamoeba histolytica Dog appearance Amoebiasis, Dog caused by Balantidium coli 90052.doc -12- 200423950 Ciliariasis and so on. For birds, there are domestic duck viral enteritis caused by Duck virus enteritis virus, turkey coronavirus enteritis caused by turkey coronavirus, avian rotavirus disease caused by rotavirus, and hemorrhagic enteritis virus.
之火雞出血性腸炎、雞小病毒引發之雞小病毒病、火雞星 狀病毒引發之火雞星狀病毒病、Yersinia pseudotuberculosis引 發之鳥類假性結核、Salmonella pollorum引發之雛白痛（、 Salmonella gallinarum 引發之雞斑療傷寒、Salmonella typhimurium、S. enteritidis、S· sofia、S. thompson、S. blockley、 S· heidelberg、S. infantis等引發之雞副傷寒及雞沙氏桿菌症、 Salmonella arizonae引發之火雞亞利桑那菌症、Campylobacter jejuni引發之鳥彎曲菌症、Clostridium colinum、C· perfringens、 C. septicum、C. botulinum引發之家禽芽胞梭菌症、Histomonas meleagridis引發之黑頭病、Cryptosporidium meleagridis、C· baileyi 引發之雞隱胞子蟲症等。於人類則有輪狀病毒所引起之輪狀病毒 症、Salmonella typhimurium、S. enteritidis、S. typhi、S. paratyphi 等引發之沙氏桿菌症、Shigella sonnei引發之赤痢、Vibrio parahaemolyticus、V· mimicus、V· fluvialis、V· hollisae等引發之 弧菌症、Vibrio cholerae引發之霍亂、Aeromonas hydrophila引發 之產氣單胞菌症、Plesiomonas shigelloides 引發之 Plesiomonas 症、Clostridium perfringens引發之威爾斯症、Bacillus cereus 引發之仙人掌桿菌症、金黃色葡萄球菌引發之金黃色葡萄 球菌症、輪狀病毒引發之輪狀病毒症、腸病毒引發之腸病 毒症、腺病毒引發之腺病毒症、流行性感冒病毒引發之流 行性感冒、諾沃克病毒引發之小型球形病毒症、 90052.doc -13- 200423950Turkey hemorrhagic enteritis, chicken parvovirus caused by chicken parvovirus, turkey astrovirus disease caused by turkey astrovirus, bird pseudotuberculosis caused by Yersinia pseudotuberculosis, white pain caused by Salmonella pollorum (, Salmonella gallinarum-induced typhoid fever, Salmonella typhimurium, S. enteritidis, S. sofia, S. thompson, S. blockley, S. heidelberg, S. infantis, etc. caused by chicken paratyphoid and salmonella, Salmonella arizonae Turkey Arizona mycosis, Campylobacter jejuni caused by Campylobacter bird, Clostridium colinum, C. perfringens, C. septicum, C. botulinum caused by clostridium poultry, blackhead caused by Histomonas meleagridis, Cryptosporidium meleagridis, C · Cryptosporidiosis caused by baileyi, etc. In humans, rotavirus caused by rotavirus, Salmonella typhimurium, S. enteritidis, S. typhi, S. paratyphi, etc. are caused by Sarcoidosis, Shigella sonnei, etc. Akabane, Vibrio parahaemolyticus, V. mimicus, V. fluvialis, V. holl Vibrio caused by isae, cholera caused by Vibrio cholerae, aeromonas caused by Aeromonas hydrophila, Plesiomonas caused by Plesiomonas shigelloides, Wels disease caused by Clostridium perfringens, cactus caused by Bacillus cereus, gold Staphylococcus aureus caused by Staphylococcus aureus, rotavirus caused by rotavirus, enterovirus caused by enterovirus, adenovirus caused by adenovirus, influenza caused by influenza virus, Norwalk virus Caused by small spherical virus disease, 90052.doc -13- 200423950
Campylobacter jejuni等引發之彎曲菌症、大腸桿菌引發之大 腸桿菌症、Yersinia enterocolitica、Y· pseudotuberculosis引發之 耶氏桿菌症、阿米巴引發之阿米巴赤痢等。 以上述腹瀉症病原體對鳥類進行免疫時，根據實際情況 可使用傅氏完全佐劑（FCA)、傅氏不完全佐劑（FIA)等佐 剤。免疫主要藉由靜脈内、皮下、肌肉、腹腔内注射方式 貫施’也可藉由經口投藥、點鼻、點眼等方式實施。又， 免疫間隔沒有特別限定，間隔數日至數週，實施丨〜丨〇次免 疫。通常，初次免疫起數週内，可從蛋、特別蛋黃中獲得 對所投藥抗原具有特效之抗體。接種抗原之使用適宜量以 蛋白質質量計算為0.01〜10 mg。 蛋之k體價可利用酵素免疫測定法（ELIS A)、放射性免疫 測疋法、减集抗體法或中和反應等測試，藉由間隔2週左右 測疋免疫後之抗體價，可跟蹤抗體價之變化情況。通常， y ^ 4個月獲得高抗體價。其間，若發現免疫後抗體價減 守可藉由間隔適當時間適當追加免疫而維持較高抗體 價。 本發明使用上述免疫之雞蛋或其處理物製造抗腹瀉症組 口物W樂組合物、家畜飼料及食品等。本發明中，蛋之 處理物右包含以鳥類免疫所使用之腹濕症病原體為抗原所 產f之抗體即可，沒有特別限定。例如，免疫蛋之全蛋、 蛋’、及蛋白，該等蛋液及溶液；或者蛋液以丙醇或氯仿萃 =得之萃取物。從所含有抗體之量考慮，優選包含蛋黃 也包3利用噴霧乾燥法或冷凍乾燥法所得之粉末。 90052.doc -14- 200423950 ^包含利用㈣基甲基纖維素鄰苯二甲酸醋、聚乙二醇、 葡萄聚糖硫酸鹽、丙醇、乙醇及己炫等有機溶劑除去蛋普 中蛋黃脂質成分後粉末化所得物。也包含該粉末之糊狀= 2狀物。再者’本發明中蛋之處理物也可指利用硫酸銨鹽 ^鹽析、低溫乙醇沈㈣、離子交換層析法、膠 7、親和層析法等f知方法從蛋中精製所得之抗體本身: 物：：：上所述製得之抗體稱為雞蛋抗體。為提高處理 :〖’優選減菌後之全蛋蛋液或蛋黃蛋液經噴霧乾 、木或冷凍乾燥所得之粉末。 本發明發現’於已患腹㈣之動物投藥上述所得 處理物之同時蔣鱸 w ^ — 了“活囷劑，可明顯減輕腹瀉症狀及其伴隨 品吐及體重減少等症狀。 衍^明中活菌劑係活的細菌、酵母及真菌，可利用本技 细d:方法製造。本發明中適於作為活菌劑使用之 屬細菌ί之=限定，包含例如屬於乳酸桿菌屬、鏈球菌 菌等之有4二:屬於芽胞梭菌屬、雙叉乳酸桿菌屬細 孢浐…一 s及屬於％球囷屬、乳酸球菌屬、芽 孢#囷屬細菌等。作為菌 才 畢赤酵母菌屬之酵母」: 包含屬於念珠菌屬、 包人…厘 作為活囷劑之真菌，沒有特別限定， 匕a例如屬於麴菌屬、 優選使用屬m 囷。本發明中，特別 ” 馱桿菌屬、鏈球菌屬、乳酸球菌屬、雔叉 乳酸桿菌屬、芽胞梭菌屬 欠囷4雙又 屬於人秦# 干囷屬、腸球菌屬之細菌； 屬於心珠囷屬之酵母；屬囷 等活菌劑可單獨使用，亦可幾種混合=困屬之真菌。該 90052.doc 200423950 作為本發明中優選使用之活菌劑，更為具體地可例舉酪 蛋白乳酸桿菌、嗜酸乳桿菌、涎鏈球菌、加氏乳酸桿菌 (Lactobacillus gasseri)、乳酸桿菌屬酵母菌（Lactobacillus Fermentum)、瑞 士乳酸桿菌、Lactobacillus yogurty、嗜熱 性鏈球菌、乳鏈球菌、雙岐桿菌、Bifidobacterium breve、 嬰兒雙岐桿菌、雙叉乳酸桿菌、Bifidobacterium pseudolongum、嗜熱雙岐桿菌（Bifidobacterium thermophilum)、 丁酸梭菌（Clostridium butyricum)、仙人掌桿菌、乳酸球菌、 腸球菌、白色念珠菌、黑麴菌、麴菌、酵母菌等。優選例 如嗜酸乳桿菌與雙岐桿菌與乳酸球菌；涎鏈球菌與嗜熱雙 岐桿菌；嗜熱性鏈球菌與Bifidobacterium pseudolongum組 合使用。又，對於病毒，特別輪狀病毒及小病毒為病原體 之腹瀉症，適於使用有益腸内細菌、特別雙叉乳酸桿菌屬 細菌作為活菌劑；對於細菌為病原體之腹瀉症，適於使用 乳酸桿菌屬細菌作為活菌劑。特別地，對於大腸桿菌屬細 菌為病原體之腹瀉症，適於使用芽孢桿菌屬細菌作為活菌 劑；對於沙氏桿菌屬細菌為病原體之腹瀉症，適宜使用有 益腸内細菌、特別是雙叉乳酸桿菌屬細菌作為活菌劑；對 於芽胞梭菌屬細菌為病原體之腹瀉症，適宜使用乳酸菌、 特別是乳酸桿菌屬細菌作為活菌劑。再者，對於原蟲、特 別隱胞子蟲屬之原蟲為病原體之腹瀉症，適宜使用乳酸 菌、特別是乳酸桿菌屬細菌作為活菌劑。 本發明之抗腹瀉症組合物中，對腹瀉症病原體免疫之鳥 類所產蛋或其處理物與活菌劑之混合比例，沒有特別限 90052.doc -16- 200423950 定，通常為對應於含有！ §抗體之蛋、活菌劑為1〇〜1〇11個之 比例，優選使用對應於！ g抗體、活菌劑為1〇4〜1〇9個之比例。 如上所得抗腹瀉症組合物通常包含〇 〇〇1〜1〇〇重量％，優 選0.01〜10重量％比例之上述蛋或其處理物與活菌劑之混合 物，可製成溶液、粉末、顆粒、片劑或糊狀等形式。另外， 由於該抗腹瀉症組合物對動物之腹瀉症具有預防及治療效 果’可作為抗腹瀉症醫藥組合物使用。又，藉由於家畜詞 料及食品中添加抗腹瀉症組合物，對未患腹瀉症之動物亦 有預防效果。於醫藥組合物、家畜飼料及食品，抗腹瀉症 組合物之添加比例通常為〇·〇〇!〜！⑽重量％，優選〇〇1〜1〇重 量 〇/〇。 片劑、顆粒劑、散劑、膠囊劑、液劑等口服用製劑。根據 情況需要使用例如賦形劑、黏結劑、崩潰劑、潤滑光澤劑、 抗氧化劑、著色劑、矯味劑等添加劑。為防止於胃部消化 製造本發明之包含抗腹瀉症組合物之醫藥組合物時，可 直接或與慣用之添加劑混合，利用通常之製劑化方法製得 分解、長時間作用於小腸部位，可實施緩釋化，亦可包覆 習知緩釋劑。作為賦形劑，可使用例如羧甲基纖維素鈉鹽復 魚膠、輕無水矽酸、明膠、結晶纖維素、山梨聚糖醇f滑 石、糊精、澱粉、乳糖、白糖、葡萄糖、甘露醇、鋁矽酸 鎂、瑞酸氫約等。作為黏結劑’可例舉阿拉伯膠、褐藻酸 鈉、乙醇、乙基纖維素、酪蛋白鈉、羧甲基纖維素鈉 魚膠、精製水、明膠、澱粉、黃蓍膠、乳糖、羥基纖維素、 羥曱基纖維素、羥曱基丙基纖維素、聚乙烯吡咯啶酉同等。 90052.doc -17- 200423950 作為崩潰劑，可例舉羧甲基纖維素、羧甲基纖維素鈉鹽、 叛甲基纖維素妈鹽、結晶纖維素、殿粉、經丙澱粉等。作 為潤滑光澤劑，可例舉硬脂酸、硬脂酸鈣、硬脂酸鎂、滑 石、硬化油、蔗糖脂肪酸酯、蠟類等。作為抗氧化劑，可 例舉生育酚、沒子酸酯、二丁羥基甲苯（bht)、丁基羥基甲 氧苯（ΒΗΑ)、抗壞血酸等。再者，根据情況也可添加其他添 加劑或藥劑，例如抗酸劑（碳酸氫鈉、碳酸鎂、沈降碳酸鈣、 水滑石等）、胃黏膜保護劑（合成矽酸鋁、硫糖鋁、葉綠素酮 鈉等）。 作為本發明之抗腹瀉症組合物或者包含該組合物之醫藥 組合物、食品及飼料之給予對象之動物無特別限定，為患 有上述腹瀉症病原體引發之腹瀉症之動物即可，例如哺乳 動物、鳥類等。本發明之抗腹瀉症組合物適合於人、牛、 豬、馬、綿羊、山羊等哺乳動物，或雞、鶉等鳥類。 本發明說明書包含作為本申請優先權基礎之特願 2002-359632號說明書中所記載之内容。 【實施方式】 實施發明之最佳狀態 以下’列舉實施例更為詳細說明本發明，然而本發明並 非限定於該等實施例。 [實施例1] 作為活g劑之酸乳桿g謂·丨132株於刪培養基培 養、集菌後’經冷;東乾燥製成lxlG、/g之粉末。作為 劑之嗜熱雙岐桿g;CM_12G7株於MRS培養基培養、 90052.doc -18- 200423950 後，經冷凍乾燥作成lxl01G個/g之粉末。作為活菌劑之 Bifidobacterium pseudolongum JCM-1205 株於 MRS 培養基 培養、集菌後，經冷凍乾燥作成lx 101G個/g之粉末。作為活 菌劑之凝結芽孢桿菌（Bacillus coagulans) JCM-2257株於普 通魚膠培養基培養、集菌後，經冷凍乾燥作成lxl01G個/g 之粉末。 [實施例2] 牛輪狀病毒島根株以赤毛猴腎細胞（MA-104)培養、超離 心採集病毒粒子。含有該病毒109TCID5G之溶液與傅氏佐劑 乳化後，藉由於12週齡之母雞之左右胸肌各注射1 ml，實 施第一次免疫。同樣地，6週後實施第二次免疫。自第二次 免疫起兩週後，利用中和反應測定該雞血液中抗輪狀病毒 抗體之抗體價（Arch. Virol·，69:49-60, 1981)為 6400倍。該雞 所產蛋之抗體價為6400倍。該抗體價持續4個月。採集該 蛋，經喷霧乾燥製成全蛋粉末。該粉末之抗體價為6400倍。 使用混合有該抗牛輪狀病毒抗體與活菌劑之飼料，檢查 其對牛輪狀病毒試驗感染（Archives Virology 138:143-148，1994) 之效果。受試動物為1日齡之新生牛，10日内分別或混合投 藥含有3200倍抗體價抗體之牛奶與含有作為活菌劑之嗜熱 雙岐桿菌JCM-1207株lx 106個之牛奶。另外，相同方法投藥 活菌劑IxlO6個與上述1/10量或1/100量之抗體之混合物。設 定為牛輪狀病毒島根株lxl01GTCID5〇感染，每群4頭共6群。 即，為抗體單獨（3200倍）投藥群、活菌劑單獨（106個/g)投藥 群、活菌劑106個/g分別與抗體3200倍、320倍、32倍混合投 90052.doc -19- 200423950 藥群。另外還有無任何投藥之對照群。又，感染後觀察期 為10天。認為抗體可保護腸導黏膜，阻止輪狀病毒附著於 黏膜。認為活菌劑可調整腸内菌叢、改善腹瀉症狀。檢查 内容為每日臨床觀察、病毒排泄期及體重增加測定，結果 示於表1。結果表明抗體與活菌劑混合投藥群於減輕腹瀉及 嘔吐、減輕病毒排泄及增加體重方面，表現明顯改善之效 果。又，活菌劑與1/100量之抗體之混合物投藥群也可得到 與抗體單獨投藥群同等以上之效果。 表1抗牛輪狀病毒抗體與活菌劑 試驗群 對照群 抗體 3200倍 活菌劑 106個 抗體+活菌劑 3200倍+106 個 抗體+活菌劑 320倍+106 個 抗體+活菌劑 32倍+106個 死亡率（％) 0 0 0 0 0 0 腹瀉發病率（％) 100 100 100 0 25 50 臨床得分合計 13 12 12 01 31 12 感染10天中之增 加體重率（％) -8 -1 1 -21 42 22 01 病毒排泄期（天） 8 51 7 22 31 41 90052.doc -20- 1 : P<0.05、2 : P<0.01 [實施例3] 2 狗小病毒Cp83016株以貓腎細胞（CRFK)培養、超離心採 集病毒粒子。含有該病毒l〇9TCID5G之溶液與傅氏佐劑乳化 後，藉由於12週齡之母雞之左右胸肌各注射1 ml，實施第 一次免疫。同樣地，6週後實施第二次免疫。自第二次免疫 起兩週後，利用中和反應測定該雞血液中抗小病毒抗體之 抗體價（J· Vet. Med· Sci·，60:973-974, 1998)為 50,000倍。該 雞所產蛋之抗體價為50,000倍。該抗體價持續4個月。採集 該蛋，經喷霧乾燥製成全蛋粉末。該粉末之抗體價為51，200 200423950 倍。 使用此合有該抗狗小病毒抗體與活菌劑之飼料，檢查其 對狗小病毒試驗感染（J· Virology，:4506_45 i 3，1994)之效 果。受試動物為3週齡之幼犬，14日内分別或混合投藥含有 25,600倍抗體價抗體之牛奶與含有作為活菌劑之Campylobacter jejuni, etc. caused Campylobacter disease, coliform caused by E. coli, Yersinia enterocolitica, Y. pseudotuberculosis caused by Yersinia disease, amoebic caused by amoeba, etc. When immunizing birds with the above-mentioned diarrhea pathogens, adjuvants such as Fuller's complete adjuvant (FCA) and Fourier's incomplete adjuvant (FIA) can be used according to the actual situation. Immunization is mainly performed by intravenous, subcutaneous, intramuscular, or intraperitoneal injection. It can also be administered by oral administration, nose, eye, etc. In addition, the immunization interval is not particularly limited, and the immunization is performed 丨 to 丨 0 times at intervals of several days to several weeks. Usually, within a few weeks from the initial immunization, antibodies specific for the administered antigen can be obtained from eggs, especially egg yolks. The appropriate amount of inoculated antigen is 0.01 to 10 mg based on the protein mass. The k value of eggs can be tested by enzyme immunoassay (ELIS A), radioimmunoassay, subtractive antibody method or neutralization reaction. By measuring the antibody value after immunization about 2 weeks apart, the antibody can be tracked Price changes. Usually, y ^ 4 months get high antibody prices. In the meantime, if it is found that the antibody price decreases after immunization, it is possible to maintain a higher antibody price by appropriate additional immunization at appropriate intervals. In the present invention, the above-mentioned immunized eggs or processed products thereof are used to produce an anti-diarrheal group mouthpiece composition, a livestock feed, food, and the like. In the present invention, the processed product of the egg may include an antibody produced by using an ascites pathogen that is used for bird immunization as an antigen, and is not particularly limited. For example, whole eggs, eggs, and proteins of immune eggs, such egg liquids and solutions; or egg liquids are extracted with propanol or chloroform. From the viewpoint of the amount of the antibody contained, it is preferable to include a powder obtained by spray drying or freeze drying including egg yolk. 90052.doc -14- 200423950 ^ Contains the removal of egg yolk lipids from egg pudding using organic solvents such as methyl methyl cellulose phthalate, polyethylene glycol, glucosan sulfate, propanol, ethanol, and hexane. After powdering the resultant. Contains paste of this powder = 2 substances. Furthermore, the processed product of eggs in the present invention may also refer to antibodies purified from eggs using ammonium sulfate, salting out, low-temperature ethanol precipitation, ion exchange chromatography, gel 7, affinity chromatography and other known methods. In itself: The antibody :: The antibody prepared above is called egg antibody. In order to improve the treatment: ['Preferred powder obtained by sterilizing whole egg liquid or egg yolk liquid by spray-drying, wood or freeze-drying. According to the present invention, it is found that while the above-obtained treatment is administered to an animal that has suffered from diarrhea, the “perch” can be used to significantly reduce the symptoms of diarrhea and its accompanying vomiting and weight loss. Bacterial agents are live bacteria, yeasts, and fungi, which can be produced by using the d: method of this technology. In the present invention, bacteria belonging to the genus suitable for use as a viable agent are limited, and include, for example, those belonging to the genus Lactobacillus, Streptococcus, etc. Among them are 42: belonging to Clostridium spp., Lactobacillus sp. Spores ... 1 s and bacteria belonging to the genus Streptococcus, Lactobacillus, Spore # 囷, etc. As yeasts of the genus Pichia ”: It includes fungi belonging to the genus Candida, including humans, etc., which are used as live tinctures, and are not particularly limited. For example, the genus a belongs to the genus Pleurotus, and the genus m 囷 is preferably used. In the present invention, especially "Pseudomonas, Streptococcus, Lactobacillus, Lactobacillus, Clostridium sp Saccharomyces spp .; Saccharomyces spp. Can be used alone, or several kinds of mixed = fungal spp. The 90052.doc 200423950 is preferably used as a saccharomyces spp. In the present invention, and more specifically can be exemplified Lactobacillus protein, Lactobacillus acidophilus, Streptococcus salivarius, Lactobacillus gasseri, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus yogurty, Streptococcus thermophilus, Streptococcus lactis, Bifidobacterium Bacillus, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium lactis, Bifidobacterium pseudolongum, Bifidobacterium thermophilum, Clostridium butyricum, Cactus, Lactobacillus, Enterococcus, Candida albicans, Black fungus, Pseudomonas, yeast, etc. Preferred are, for example, Lactobacillus acidophilus and Bifidobacterium and lactococcus; Streptococcus salivarius and Thermophilic Bifidobacterium; Thermophilic Streptococcus combined with Bifidobacterium pseudolongum. Also, for viruses, particularly rotaviruses and parvoviruses as pathogens of diarrhea, it is suitable to use beneficial intestinal bacteria, especially Bifidobacterium bacteria Live bacterial agent; for diarrhea with bacteria as pathogen, it is suitable to use lactobacillus bacteria as live bacteria. In particular, for diarrhea with E. coli bacteria as pathogen, it is suitable to use bacillus bacteria as live bacteria; For diarrhea of Salmonella bacteria as pathogens, it is suitable to use beneficial intestinal bacteria, especially Bifidobacterium bacteria as living bacteria; for diarrhea of Clostridium bacteria as pathogens, lactic acid bacteria, especially Lactobacillus bacteria are used as viable agents. Furthermore, for protozoa, especially those of the genus Cryptosporidium, which are pathogens, lactic acid bacteria, especially bacteria of the genus Lactobacillus, are suitable as viable agents. Anti-diarrhea of the present invention Eggs, or their preparations and live bacteria, from birds immune to diarrheal pathogens The mixing ratio is not particularly limited to 90052.doc -16- 200423950, but it usually corresponds to the ratio of 10 to 1011 eggs containing eggs and live bacteria, and it is preferable to use the antibodies corresponding to! G antibodies and live bacteria. The agent is in a ratio of 104 to 109. The anti-diarrheal composition obtained as above usually contains 0.001 to 100% by weight, preferably 0.01 to 10% by weight of the above-mentioned egg or its processed material and live bacteria. Mixtures of agents can be made into solutions, powders, granules, tablets or pastes. In addition, since the anti-diarrheal composition has a preventive and therapeutic effect on diarrhea in animals, it can be used as an anti-diarrheal pharmaceutical composition. In addition, by adding anti-diarrheal composition to livestock words and food, it has a preventive effect on animals without diarrhea. The addition ratio of the anti-diarrheal composition to the pharmaceutical composition, livestock feed and food is usually 〇〇〇〇! ~! ⑽% by weight, preferably 0.001 to 10% by weight 〇 / 〇. Oral preparations such as tablets, granules, powders, capsules, and liquids. Additives such as excipients, binders, disintegrating agents, lubricating gloss agents, antioxidants, colorants, flavoring agents, etc. are used as required. In order to prevent digestion in the stomach, the pharmaceutical composition containing the anti-diarrheal composition of the present invention can be directly or directly mixed with conventional additives, and can be decomposed by a common preparation method, and can be applied to the small intestine for a long time. Slow release, can also be coated with conventional slow release agents. As the excipient, for example, carboxymethyl cellulose sodium salt fish gelatin, light anhydrous silicic acid, gelatin, crystalline cellulose, sorbitan f-talc, dextrin, starch, lactose, white sugar, glucose, mannitol can be used. , Magnesium aluminosilicate, hydrogen arsenate and so on. Examples of the binder include gum arabic, sodium alginate, ethanol, ethyl cellulose, sodium casein, sodium carboxymethyl cellulose, fish gelatin, purified water, gelatin, starch, tragacanth, lactose, and hydroxycellulose. , Hydroxymethyl cellulose, hydroxymethyl propyl cellulose, polyvinylpyrrolidine, the same. 90052.doc -17- 200423950 As the disintegrating agent, carboxymethyl cellulose, carboxymethyl cellulose sodium salt, methylcellulose mother salt, crystalline cellulose, sodium powder, trimethyl starch, etc. may be mentioned. As the lubricating gloss agent, stearic acid, calcium stearate, magnesium stearate, talc, hardened oil, sucrose fatty acid esters, waxes, and the like can be exemplified. Examples of the antioxidant include tocopherol, gallate, dibutylhydroxytoluene (bht), butylhydroxymethoxybenzene (ΒΗΑ), and ascorbic acid. In addition, other additives or agents can be added according to the situation, such as antacids (sodium bicarbonate, magnesium carbonate, precipitated calcium carbonate, hydrotalcite, etc.), gastric mucosal protective agents (synthetic aluminum silicate, sucralfate, chlorophyllone) Sodium, etc.). The animal to be administered as the anti-diarrheal composition or the pharmaceutical composition, food and feed containing the composition of the present invention is not particularly limited, and it can be an animal suffering from diarrhea caused by the above-mentioned pathogens of diarrhea, such as mammals, Birds, etc. The anti-diarrhea composition of the present invention is suitable for mammals such as humans, cattle, pigs, horses, sheep, goats, or birds such as chickens and quails. The present specification includes the contents described in Japanese Patent Application No. 2002-359632, which is the basis of the priority of the present application. [Embodiment] Best Mode for Carrying Out the Invention The following examples are used to illustrate the present invention in more detail, but the present invention is not limited to these embodiments. [Example 1] 132 g of yoghurt as a live g agent was cultured and cultured in a delete medium, and the strain was' cold; dried to make a powder of lxlG, / g. As an agent, thermophilic bifidus g; CM_12G7 strain was cultured in MRS medium, 90052.doc -18- 200423950, and then freeze-dried to obtain lxl01G / g powder. Bifidobacterium pseudolongum JCM-1205 strain as a viable bacterial agent was cultured in MRS medium, collected bacteria, and freeze-dried to make a powder of lx 101G / g. Bacillus coagulans JCM-2257 strain, which is used as a viable bacterial agent, was cultured in ordinary fish gelatin medium, collected, and freeze-dried to make a powder of lxl01G / g. [Example 2] Bovine rotavirus island root strains were cultured with red wool monkey kidney cells (MA-104), and virions were collected by ultra-centrifugation. After emulsification of the solution containing the virus 109TCID5G and Fourier's adjuvant, the first immunization was performed by injecting 1 ml of left and right breast muscles of a 12-week-old hen. Similarly, a second immunization was performed after 6 weeks. Two weeks after the second immunization, the antibody value of the anti-rotavirus antibody in the blood of the chicken was determined by a neutralization reaction (Arch. Virol., 69: 49-60, 1981) to be 6400 times. The egg produced by this chicken is 6400 times more expensive. The antibody value lasted for 4 months. The eggs were collected and spray-dried to obtain whole egg powder. The powder has an antibody value of 6400 times. A feed mixed with the anti-bovine rotavirus antibody and a live bacterial agent was used to examine its effect on the experimental infection of bovine rotavirus (Archives Virology 138: 143-148, 1994). The test animals were newborn cows at 1 day of age. Milk containing 3200 times the antibody value of antibodies and milk containing 106 cells of Bifidobacterium thermophilus JCM-1207 strain as live bacteria were administered separately or mixed within 10 days. In addition, the same method was used to administer a mixture of IxlO6 viable bacterial agent and the above-mentioned antibody in an amount of 1/10 or 1/100. It was assumed that the infection was caused by bovine rotavirus island root strain lxl01GTCID50, with 4 heads in each group and a total of 6 groups. That is, for the antibody alone (3200 times) administration group, the live bacteria agent alone (106 pcs / g) administration group, and the live bacteria agent 106 pcs / g, respectively, and the antibody were administered at 3200 times, 320 times, and 32 times, respectively. -200423950 Medicine group. There was also a control group without any administration. The observation period after infection was 10 days. Antibodies are thought to protect the intestinal mucosa and prevent rotavirus from attaching to the mucosa. It is believed that live bacteria can adjust the intestinal flora and improve the symptoms of diarrhea. The examination contents are daily clinical observation, viral excretion period and weight gain measurement. The results are shown in Table 1. The results showed that the mixed administration of the antibody and the live bacterial agent showed a significant improvement in reducing diarrhea and vomiting, reducing viral excretion, and increasing weight. In addition, the administration of a mixture of a viable bacterial agent and an antibody in an amount of 1/100 can achieve the same effect or more as that of the administration of the antibody alone. Table 1 Antibovine rotavirus antibodies and live bacteria test group control group antibodies 3200 times live bacteria agent 106 antibodies + live bacteria agent 3200 times + 106 antibodies + live bacteria agent 320 times + 106 antibodies + live bacteria agent 32 +106 deaths (%) 0 0 0 0 0 0 Incidence of diarrhea (%) 100 100 100 0 25 50 Total clinical score 13 12 12 01 31 12 Weight gain rate (%) within 10 days of infection -8- 1 1 -21 42 22 01 Virus excretion period (days) 8 51 7 22 31 41 90052.doc -20- 1: P < 0.05, 2: P < 0.01 [Example 3] 2 Canine parvovirus Cp83016 strain is cat kidney Cells (CRFK) were cultured and virions were collected by ultracentrifugation. After emulsifying the solution containing the virus 109TCID5G and Fourier's adjuvant, the first immunization was performed by injecting 1 ml of left and right breast muscles of a 12-week-old hen. Similarly, a second immunization was performed after 6 weeks. Two weeks after the second immunization, the antibody value (J. Vet. Med. Sci., 60: 973-974, 1998) of the antibody against parvovirus in the blood of the chicken was measured by a neutralization reaction to be 50,000 times. The antibody price of the eggs laid by this chicken is 50,000 times. The antibody value lasted for 4 months. The eggs were collected and spray-dried to obtain whole egg powder. The antibody value of this powder is 51,200 200423950 times. The feed containing the anti-canine parvovirus antibody and a live bacterial agent was used to check its effect on dog parvo test infection (J. Virology, 4506_45 i 3, 1994). The test animals were puppies of 3 weeks of age. Within 14 days, milk containing 25,600 times the antibody value of antibodies and food containing live bacteria were administered separately or in combination.
Bifidobacterium pseudolongum JCM-1205 株 lxl〇6 個之牛 奶。另外’相同方法投藥活菌劑1 χ丨〇6個與上述丨/丨〇量或 1/10〇ϊ之抗體之混合物。設定為狗小病毒Cp83〇16株6χ1〇8 TCIDsg感染’每群3頭共6群。即，試驗群為抗體單獨（25,600 倍）投藥群、活菌劑單獨（1〇6個/g)投藥群、活菌劑1〇6個仏分 別與抗體25,600倍、2560倍、256倍混合投藥群。另外還有 無任何投藥之對照群。又，感染後觀察期為14天。認為抗 體可保護腸導黏膜、阻止小病毒附著於黏膜。認為活菌劑 可凋正腸内菌叢、改善腹瀉症狀。檢查内容為每日臨床觀 察、病毒排泄期及體重增加測定，結果示於表2。結果表明 抗體與活菌劑混合投藥群於減輕腹瀉及嘔吐、減輕病毒排 泄及增加體重方面，表現明顯改善之效果。又，活菌劑與 1/100里之抗體之混合物投藥群也可得到與單獨投藥抗體 群同等以上之效果。 表2抗狗小病毒抗體與活菌劑 ---—---^ 試驗群 腹瀉發^~ (%) 對照群 100 ~Μ~ 丨25600倍 67 活菌劑 1〇6個 100~~ --——-~~ 抗體+活菌劑 _^60〇^1〇β個— 0 抗體+活菌劑 2560倍+106 個 0 抗體+活菌劑 256 倍+106 個 33 股^舄病期 (天） ---—.___ 3 2 Ί 2 --—--- 〇** —~------ 1 90052.doc 21 200423950 S*床得分合計 7 3 5 2* 感染14天 增加體重(kg) -0.1 0.2** 0.1 0.3** 0.2** 〇.2** 病毒排泄期 (天） 12 7** 10 6** 6** 7** * : Ρ<0·05、** : Ρ<〇·〇1 [實施例4] 來自豬大腸桿菌8199株（具有F18附著纖毛）於MINCA培 養基培養、超離心採集菌體，藉由加熱至6〇〇C提取F18附著 纖毛（Vet· Microbiol·，45:281-295, 1995)。含有該F18附著纖 毛〇·5 mg/ml之溶液與傅氏佐劑乳化後，藉由於12週齡之母 雞之左右胸肌各注射1 ml，實施第一次免疫。同樣地，6週 後實施第二次免疫。自第二次免疫起兩週後，利用凝集抗 體反應測定該雞血液中F18纖毛抗體之抗體價為256〇倍。該 矣隹所產蛋之抗體價為2560倍。該抗體價持續4個月。採集該 蛋，經噴霧乾燥製成全蛋粉末。該粉末之抗體價為256〇倍。 使用混合有該抗F18纖毛抗體與活菌劑之飼料，檢查其對 豬大腸桿菌實驗感染（j. Vet Med Sci.，59:917_921，1997) 之效果。受試動物為4週齡之斷奶豬，10日内投藥混合有1〇 倍抗體價抗體與作為活菌劑之凝結芽孢桿菌（Badiius coagulans) jCM_1132株1χ1〇4個/g中一種或二者皆有之飼 料。另外，相同方法投藥活菌劑1><1〇4個々與上述ι/ι〇量或 1/100量之抗體之混合物。設定為來自緒大腸桿菌株 1x10個感* ’母群10頭共6群。即，試驗群為抗體單獨（⑺ 倍）投藥群、活菌劑單獨投藥⑽4個/g)群、活菌劑1〇4個^分 別與抗體1G倍、！倍、G1倍混合投藥群。另外還有無任何 90052.doc -22- 200423950 投藥之對照群。又，感染後觀察期為10天。認為抗體可保 護腸導黏膜、阻止具有F18纖毛之大腸桿菌附著於黏膜。認 為活菌劑可調整腸内菌叢、改善腹瀉症狀。檢查内容為每 曰臨床觀察、攻擊大腸桿菌分離率之減輕及體重增加測 定，結果示於表3。結果表明抗體與活菌劑混合投藥群於減 輕腹瀉及嘔吐、減輕攻擊大腸桿菌分離率及增加體重方 面，表現明顯改善之效果。又，活菌劑與1 /10 0量之抗體之 混合物投藥群也可得到與單獨投藥抗體群同等以上之效 果0 表3抗來自豬大腸桿菌抗體與活菌劑 試驗群 對照群 抗體 10 活菌劑 104個 抗體+活菌劑 10倍+104個 抗體+活菌劑 1倍+104個 抗體+活菌劑 0.1 倍+104 個 死亡率（％) 0 0 0 0 0 0 臨床得分合計 13 12 12 01 4氺 6 感染第3天之臨 床得分(％) 40 20 30 101 101 101 感染第3天攻擊 菌分離率（％) 90 70 80 302 401 50 感染10天後體重 增加率（％) 52 52 52 802 752 60 90052.doc -23- 1 : Ρ<0·05、2 : Ρ<0·01 [實施例5]Bifidobacterium pseudolongum JCM-1205 strain lx106 milk. In addition, the same method was used to administer a mixture of 1 x 6 of live bacteria and the above-mentioned antibody in an amount of 1/100 or 1/10. Canine parvovirus Cp83〇16 strain 6χ108 TCIDsg infection 'was assumed to be 6 groups of 3 heads per group. That is, the test group was the antibody (25,600 times) administration group alone, the live bacteria agent alone (106 units / g) administration group, and the live bacteria agent 106 groups, and the antibodies were mixedly administered with the antibodies 25,600 times, 2560 times, and 256 times, respectively. group. There was also a control group without any administration. The observation period after infection was 14 days. Antibodies are thought to protect the intestinal mucosa and prevent small viruses from attaching to the mucosa. It is believed that live bacteria can wither the intestinal flora and improve the symptoms of diarrhea. The examination contents were daily clinical observation, viral excretion period and weight gain measurement. The results are shown in Table 2. The results showed that the mixed administration of the antibody and the live bacterial agent showed a significant improvement in reducing diarrhea and vomiting, reducing viral excretion, and increasing weight. In addition, the administration of a mixture of a live bacterial agent and an antibody in a ratio of 1/100 can achieve the same effect or more as that of the antibody group administered alone. Table 2 Anti-canine parvovirus antibodies and live bacteria agents -------- ^ Test group diarrhea ^ ~ (%) Control group 100 ~ M ~ 丨 25600 times 67 Live bacteria agent 100 100 ~~- ——- ~~ Antibody + Live Bacteria_ ^ 60〇 ^ 1〇β A — 0 Antibody + Live Bacteria 2560 times + 106 0 0 Antibody + Live Bacteria 256 times + 106 33 shares ) -----.___ 3 2 Ί 2 ------ 〇 **-~ ------ 1 90052.doc 21 200423950 S * bed score total 7 3 5 2 * 14 days of weight gain after infection ( kg) -0.1 0.2 ** 0.1 0.3 ** 0.2 ** 〇.2 ** Virus excretion period (days) 12 7 ** 10 6 ** 6 ** 7 ** *: Ρ < 0 · 05, **: P < 〇 · 〇1 [Example 4] From Swine Escherichia coli 8199 strain (with F18 attached cilia) was cultured in MINCA medium, the cells were collected by ultracentrifugation, and F18 attached cilia (Vet · Microbiol were extracted by heating to 600 ° C). ·, 45: 281-295, 1995). After the solution containing the F18 attached cilia 0.5 mg / ml was emulsified with Fourier's adjuvant, the first immunization was performed by injecting 1 ml of left and right breast muscles of a 12-week-old hen. Similarly, a second immunization was performed after 6 weeks. Two weeks after the second immunization, the antibody value of the F18 ciliary antibody in the blood of the chicken was measured by agglutination antibody response to be 256-fold. The antibody value of eggs produced by this tadpole is 2560 times. The antibody value lasted for 4 months. The eggs were collected and spray-dried to obtain whole egg powder. The powder has an antibody value of 2560 times. A feed mixed with the anti-F18 ciliary antibody and a live bacterial agent was used to check its effect on experimental infection of swine E. coli (j. Vet Med Sci., 59: 917_921, 1997). The test animal was a 4-week-old weaned pig. Within 10 days of administration, 10 times the antibody valence antibody was mixed with Badiius coagulans jCM_1132 strain 1 × 104 / g or both. Of feed. In addition, the same method was used to administer a mixture of the live bacterial agent 1 > < 104 gadolinium and the antibody in the amount of 1/100 or 1/100. It was set as 6 groups of 10 heads from the E. coli strain * 'mother group with 10 heads. In other words, the test group is the antibody alone (倍 times) administration group, the live bacteria agent alone (⑽4 / g) group, and the live bacteria agent 104, respectively, 1G times the antibody, respectively! Times, G1 times mixed administration group. There is also a control group without any 90052.doc -22- 200423950 administration. The observation period after infection was 10 days. Antibodies are thought to protect the intestinal mucosa and prevent E. coli with F18 cilia from attaching to the mucosa. It is believed that live bacteria can adjust the intestinal flora and improve the symptoms of diarrhea. The examination contents are the clinical observation, the reduction of the isolation rate of challenge E. coli, and the measurement of weight gain. The results are shown in Table 3. The results showed that the mixed administration of the antibody and the live bacterial agent showed a significant improvement in reducing diarrhea and vomiting, reducing the isolation rate of attacking E. coli, and increasing body weight. In addition, the administration of a mixture of a live bacterial agent and an antibody in an amount of 1/10 can also obtain the same effect as that of the antibody group administered alone. Agent 104 antibodies + live bacteria agent 10 times + 104 antibodies + live bacteria agent 1 times + 104 antibodies + live bacteria agent 0.1 times + 104 mortality (%) 0 0 0 0 0 0 Total clinical score 13 12 12 01 4 氺 6 Clinical score on day 3 of infection (%) 40 20 30 101 101 101 Isolation rate of challenge bacteria on day 3 of infection (%) 90 70 80 302 401 50 Weight gain rate (%) after 10 days of infection 52 52 52 802 752 60 90052.doc -23- 1: P < 0 · 05, 2: P < 0 · 01 [Example 5]
Salmonella dublin 256 株於 Luria Boras 培養基中培養、超 2 離心採集菌體。含有該菌體101()個之溶液與傅氏佐劑乳化 後、藉由於12週齡之母雞之左右胸肌各注射1 ml，實施第 一次免疫。同樣地，6週後實施第二次免疫。自第二次免疫 起兩週後，利用凝集抗體法測定該雞血液中抗沙氏桿菌抗 體之抗體價為2560倍。該雞所產蛋之抗體價為2560倍。該 200423950 抗體價持續4個月。採集該蛋，經喷霧乾燥製成全蛋於 該粉末之抗體價為2560倍。 a ° 使用混合有該抗沙氏桿菌抗體與活菌劑之 > 』竹’檢查其Salmonella dublin 256 strains were cultured in Luria Boras medium and harvested by ultracentrifugation. After emulsifying the solution containing 101 () cells with Fourier's adjuvant, 12 ml of the left and right breast muscles of a 12-week-old hen were injected with 1 ml each to perform the first immunization. Similarly, a second immunization was performed after 6 weeks. Two weeks after the second immunization, the agglutination antibody method was used to determine the antibody value against the Salmonella antibody in the blood of the chicken was 2560 times. The price of the antibody produced by the chicken was 2560 times. The 200423950 antibody price lasted 4 months. The egg was collected and spray-dried to obtain whole eggs with a powder price of 2560 times. a ° Use the > "Bamboo" mixed with the anti-S. aureus antibody and live bacteria
對牛沙氏桿 實驗感染（Am. J. Vet. Res.，59:81_85 199Q 效果。受試動物為1日齡之新生牛，1〇日内分別或混人浐荠 含有2560倍抗體價之抗體之牛奶與含有作為活菌劑之嗜熱 雙岐桿菌JCM-1207株lx 106個之牛奶。另外，相同方法γ藥 活菌劑lxlO6個與上述1/10量或1/100量之抗體之混合物:設 定為來自牛Salmonella dublin 256株lxlO11個感染，每群5 頭共6群。即，試驗群為抗體單獨（256〇倍）投藥群、活菌劑 單獨（1〇6個/g)投藥群、活菌劑1〇6個/g分別與抗體256〇倍、 256倍、26倍混合投藥群。另外還有無任何投藥之對照群。 又，感染後觀察期為10天。認為抗體可保護腸導黏膜、阻 止沙氏桿菌附著於黏膜。認為活菌劑可調整腸内菌叢、改 善腹瀉症狀。檢查内容為每日臨床觀察，結果示於表4。結 果表明抗體與活菌劑混合投藥群於減輕腹瀉及嘔吐方面， 表現明顯改善之效果。又，活菌劑與1 /1 〇〇量之抗體之混合 物投藥群也可得到與單獨投藥抗體群同等以上之效果。 表4抗沙氏桿菌抗體與活菌劑 試驗群 對照群 抗體 2560倍 活菌劑 106個 抗體+活菌劑 2560倍+106 個 抗體+活菌劑 256倍+106 個 抗體+活菌劑 26倍+106個 死亡率（％) 100 40 60 20* 感染第3天之 臨床得分（％) 100 60 60 20* 30* 50 * : P<0.05、** : Ρ<〇.〇1 90052.doc -24- 200423950 [實施例6]Effect on experimental infection of bovine S. sclerotiorum (Am. J. Vet. Res., 59: 81_85 199Q. The test animal is a one-day-old newborn bovine, which contains 2560 times the antibody value of the antibody, respectively or mixed within 10 days. Milk and milk containing 106 pieces of Bifidobacterium thermophilus JCM-1207 strain as living bacteria. In addition, the same method is used as a mixture of 6 medicine-living bacterial agents 1 × 10 and the above-mentioned 1/10 or 1/100 antibody. : Set to 256 strains of bovine Salmonella dublin lxlO11 infection, 5 heads in each group, 6 groups in total. That is, the test group is the antibody alone (256 times) administration group, and the live bacterial agent alone (106 / g) administration group. 10, live bacteria agents / g were mixed with the antibodies in groups of 256 times, 256 times, and 26 times respectively. In addition, there was a control group without any administration. In addition, the observation period after infection was 10 days. It was thought that the antibodies could protect the intestines. Guide mucosa and prevent Salmonella from adhering to the mucosa. It is believed that live bacteria can adjust the intestinal flora and improve the symptoms of diarrhea. The examination content is daily clinical observations, and the results are shown in Table 4. The results show that the antibody and the live bacteria are mixed in the drug group In reducing diarrhea and vomiting, it shows a marked improvement effect. The same effect can be obtained in the administration group of the mixture of the agent and the antibody in an amount of 1/1, 000. Table 4 Anti-S. Aureus antibody and the control group of the viable bacterial test group have 106 viable bacterial agents of 2,560 times. Antibody + live bacteria agent 2560 times + 106 antibodies + live bacteria agent 256 times + 106 antibodies + live bacteria agent 26 times + 106 mortality (%) 100 40 60 20 * Clinical score on day 3 of infection (%) 100 60 60 20 * 30 * 50 *: P < 0.05, **: P < 〇.〇1 90052.doc -24- 200423950 [Example 6]
Clostridium perfringens PB6株於brucella Boras培養基中 培養、超離心採集菌體。含有該菌體lx 101G個之溶液與傅氏 佐劑乳化後，藉由於12週齡之母雞之左右胸肌各注射1 ml，實 施第一次免疫。同樣地，6週後實施第二次免疫。自第二次 免疫起兩週後，利用凝集抗體法測定該雞血液中抗體之抗 體價為640倍。該雞所產蛋之抗體價為640倍。該抗體價持 續4個月。採集該蛋，經喷霧乾燥製成全蛋粉末。該粉末之 抗體價為640倍。 使用該抗芽孢梭菌抗體與活菌劑，檢查其對鼠芽孢梭菌 貝氣感^之效果。受试動物為4週齡之b ALB/c鼠，3日内分 別或混合投藥32倍抗體價之抗體與作為活菌劑之嗜酸乳桿 菌JCM-1132株lxl〇5個。另外，相同方法投藥活菌劑1χ1〇5 個與上述1/10量或1/100量之抗體之混合物。設定為來自牛 Clostridium perfringens ΡΒ6 株 lxl〇7 個感染，每群 1〇 頭共 6 群。即，试驗群為抗體單獨（32倍）投藥群、活菌劑單獨（i 〇5 個/g)投藥群、活菌劑105個/g分別與抗體32倍、3倍、〇.3倍 此合投藥群。另，還有無任何投藥之對照群。又，感染後 之觀察期間為14天。認為抗體可保護腸導黏膜、阻止芽抱 梭菌附著於黏膜。認為活菌劑可調整腸内菌叢、改善腹篇 症狀。檢查内容為每日臨床觀察，結果示於表5。結果表明 抗體與活菌劑混合投藥料減少死亡率方面，表現明顯改 善之效果。又，活菌劑與1/1〇〇量之抗體之混合物投荜群也 可得到與單獨投藥抗體群同等以上之效果。 90052.doc -25 · 200423950 表5抗芽孢梭菌抗體與活菌劑 試驗群 對照群 抗體 32倍 活菌劑 105個 抗體+活菌劑 32倍+105個 抗體+活菌劑 3倍+105個 抗體+活菌劑 0.3 倍+105 個 死亡率（％) 80 40 60 20* 30* * : Ρ<0·05、** : Ρ<0·01 [實施例7] 令 SCID 鼠經 口感染 Cryptosporidium parvum Mito株之卵 囊、收集其糞便及腸導内容物，精製得到裂體性芽孢。含 有該裂體性芽孢0.5 mg/ml之溶液與傅氏佐劑乳化後、藉由 於12週齡之母雞之左右胸肌各注射1 ml，實施第一次免 疫。同樣地，6週後實施第二次免疫。自第二次免疫起兩週 後，利用螢光抗體法測定該雞血液中抗體之抗體價為6400 倍。該雞所產蛋之抗體價為6400倍。該抗體價持續4個月。 採集該蛋，經喷霧乾燥製成全蛋粉末。該粉末之抗體價為 6400 倍。 使用該隱孢子蟲抗體與活菌劑混合之溶液，檢查其對鼠 隱孢子蟲實驗感染之效果。受試動物為4週齡之SCID鼠，27 曰内分別或混合投藥20倍抗體價之抗體與作為活菌劑之嗜 酸乳桿菌JCM-1132株lx 105個。另外，相同方法投藥活菌劑 lxlO5個/g與上述1/10量或1/100量之抗體之混合物。 設定為 Cryptosporidium parvum Mito株 lx 107個感染，每 群6頭共6群。即，試驗群為抗體單獨（20倍）投藥群、活菌劑 單獨（105個/g)投藥群、活菌劑105個/g分別與抗體20倍、2 倍、0.2倍混合投藥群。另外還有無任何投藥之對照群。又， 感染後觀察期為27天。認為抗體可保護腸導黏膜、阻止隱 90052.doc -26- 200423950 孢子蟲附著於黏膜。認為活菌劑可調整腸内菌叢、改善腹 瀉症狀。每日臨床觀察及攻擊菌排泄量之測定作為檢查内 容，結果示於表6。結果表明抗體與活菌劑混合投藥群於減 少攻擊菌之排泄量方面，有明顯改善之效果。又，活菌劑 與1/100量之抗體之混合物投藥群中也可得到與單獨投藥 抗體群同等以上之效果。 表6抗隱孢子蟲抗體與活菌劑 由糞便卵囊之排泄量（個） 第0天 第12天 第17天 第22天 第27天 對照 0 50,000 100,000 500,000 1,000,000 抗體20倍 0 1,000* 10,000* 50,000* 500,000 活菌劑1〇5個 0 10,000 50,000 100,000 500,000 抗體+活菌劑 20倍+105個 0 100** 500** 1,000** 10,000** 抗體+活菌劑 2倍+105個 0 500** 1,000** 5,000** 50,000** 抗體+活菌劑 0.2 倍+105 個 0 1,000* 5,000* 10,000* 100,000* * : P<0.05、** : PO.01 於飼料、食品或醫藥品中混合包含本發明之抗體與活菌 劑之抗腹瀉組合物給與腹瀉症病原體之試驗感染動物時， 與抗體單獨投藥動物群相比，與活菌劑混合投藥動物群表 現出有意義之發病減輕及增加體重效果。又，於活菌劑中 即使抗體少量投藥之情形，由於乘數效應也觀測到發病減 輕及增重效果。 本說明書中所引用之全部期刊、專利及專利申請為直接 參考、引用於本說明書中。 產業應用可能性 90052.doc -27- 200423950 由於本發明之包冬p將、合—t ^ 入^11/4症抗體與$ ®劑之抗腹瀉症組 、^ ㈣體有特效，於動物之腹填症發揮比疫苗 或抗生物貝更優異之預防效果同時’可減輕腹寫症特有之 腹瀉、嘔吐及體重減輕等症狀，故可作為特效之腹瀉症預 防藥及治療藥使用。又，無諸如抗生物質之耐性菌發生之 問題。 90052.doc -28-Clostridium perfringens PB6 strain was cultured in brucella Boras medium, and the cells were collected by ultracentrifugation. After emulsifying the solution containing 1x 101G cells and Fourier's adjuvant, the first immunization was performed by injecting 1 ml of left and right breast muscles of a 12-week-old hen. Similarly, a second immunization was performed after 6 weeks. Two weeks after the second immunization, the antibody value of the antibodies in the blood of the chicken was determined to be 640 times by the agglutination antibody method. The egg price of this chicken is 640 times. The antibody price lasted for 4 months. The eggs were collected and spray-dried to obtain whole egg powder. The powder has an antibody value of 640 times. Using the anti-C. Spores antibody and live bacteria agent, the effect on the gas sensation of Clostridium spp. Was examined. The test animals were 4-week-old b ALB / c mice, and within 3 days, antibodies of 32 times the antibody value and L. acidophilus JCM-1132 strain, which is a viable bacterial agent, were administered lx105. In addition, the same method was used to administer a mixture of 1x105 live bacteria and the above-mentioned 1/10 or 1/100 antibody. It was set as 1 × 107 infections from bovine Clostridium perfringens PB6 strain, with 10 heads in each group and a total of 6 groups. That is, the test group was the antibody (32 times) administration group alone, the live bacteria alone (105 cells / g) administration group, and 105 live bacteria agents / g respectively 32, 3, and 0.3 times the antibody. This joint medicine group. There was also a control group without any administration. The observation period after infection was 14 days. Antibodies are thought to protect the intestinal mucosa and prevent Clostridium budis from attaching to the mucosa. It is believed that live bacteria can adjust the intestinal flora and improve abdominal symptoms. The examination content is a daily clinical observation, and the results are shown in Table 5. The results show that the mixed administration of antibodies and live bacteria has a significant improvement in reducing mortality. In addition, the same effect can be obtained by administering a mixture of a viable bacterial agent and an antibody in an amount of 1/100 to the antibody group alone. 90052.doc -25 · 200423950 Table 5 Anti- Clostridium antibody and live agent test group control group antibody 32 times live bacteria agent 105 antibodies + live bacteria agent 32 times + 105 antibodies + live bacteria agent 3 times + 105 Antibody + live bacteria agent 0.3 times + 105 death rate (%) 80 40 60 20 * 30 * *: P < 0 · 05, **: P < 0 · 01 [Example 7] SCID mice were orally infected with Cryptosporidium Parvum Mito's oocysts, feces and intestinal contents were collected and refined to obtain spores of spores. After the emulsification of the solution containing 0.5 mg / ml of the fission spores and Fourier's adjuvant, 1 ml of the left and right breast muscles of 12-week-old hens were injected to implement the first immunization. Similarly, a second immunization was performed after 6 weeks. Two weeks after the second immunization, the antibody value of the antibody in the blood of the chicken was determined to be 6,400 times by the fluorescent antibody method. The antibody price of the eggs laid by this chicken is 6400 times. The antibody value lasted for 4 months. The eggs were collected and spray-dried to obtain whole egg powder. The powder has an antibody value of 6400 times. Use a solution of the Cryptosporidium antibody mixed with a live bacterial agent to check its effect on the experimental infection of Cryptosporidium rat. The tested animals were SCID mice at 4 weeks of age, lx 105 of Lactobacillus acidophilus JCM-1132 strain, which was administered as a viable bacterial agent, or a 20-fold antibody, respectively, within 27 days. In addition, the same method was used to administer a mixture of lxlO5 / g and 1/1 or 1/100 antibody. Cryptosporidium parvum Mito strain was set to 107 infections with 6 heads in each group and 6 groups in total. That is, the test group was a single (20-fold) antibody-administered group, a live-bacterial agent alone (105 cells / g) -administered group, and a live-bacterial agent 105 cells / g-mixed administration group with antibodies at 20-fold, 2-fold, and 0.2-fold, respectively. There was also a control group without any administration. The observation period after infection was 27 days. Antibodies are thought to protect the intestinal mucosa and prevent cryptococcal 90052.doc -26- 200423950 spores from attaching to the mucosa. It is believed that live bacteria can adjust the intestinal flora and improve the symptoms of diarrhea. The daily clinical observation and the measurement of the amount of challenge bacteria excretion were taken as the examination contents. The results are shown in Table 6. The results show that the mixed administration of antibodies and live bacteria has a significant improvement in reducing the excretion of attacking bacteria. In addition, the same effect as that of the antibody group administered alone can be obtained in the administration group of the mixture of the live bacteria agent and the antibody in an amount of 1/100. Table 6 Excretion of anti-Cryptosporidium antibodies and live bacterial agents from fecal oocysts (a) Day 0 12 Day 17 Day 22 Day 27 Control 0 50,000 100,000 500,000 1,000,000 Antibody 20 times 0 1,000 * 10,000 * 50,000 * 500,000 live bacteria 1 105 0 10,000 50,000 100,000 500,000 antibody + live bacteria 20 times + 105 0 100 ** 500 ** 1,000 ** 10,000 ** antibody + live bacteria 2 times + 105 0 500 ** 1,000 ** 5,000 ** 50,000 ** 0.2 times antibody + live bacterial agent + 105 0 1,000 * 5,000 * 10,000 * 100,000 * *: P < 0.05, **: PO.01 in feed, food or pharmaceutical products When the anti-diarrheal composition containing the antibody of the present invention and the live bacterial agent is mixed and administered to a test-infected animal with a pathogen of diarrhea, compared with the animal group administered with the antibody alone, the animal group mixed with the live bacterium agent shows a significant reduction in disease incidence and Weight gain effect. In addition, even in the case of a small amount of the antibody administered to the live bactericide, a reduction in the incidence and weight gain were observed due to the multiplier effect. All journals, patents, and patent applications cited in this specification are directly referenced and incorporated in this specification. Industrial Application Possibility 90052.doc -27- 200423950 Due to the package of the present invention, the combination of t-^^ into the anti-diarrhea group with ^ 11/4 disease antibody and $ ® agent has special effects on the carcass Abdominal filling disease has more excellent preventive effect than vaccine or anti-bioshellfish. At the same time, it can reduce the symptoms of diarrhea, vomiting, and weight loss that are unique to abdominal dysplasia, so it can be used as a specific preventive and therapeutic agent for diarrhea. In addition, there are no problems such as the occurrence of resistant bacteria. 90052.doc -28-