TW201417846A - Method of using peracetic acid for inhibiting pathogenic microorganisms on animal farms - Google Patents
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本發明有關以過醋酸在動物養殖場中抑制病原性微生物的方法。 The present invention relates to a method of inhibiting pathogenic microorganisms in an animal farm using peracetic acid.
傳統上動物養殖業在進行水處理之應用中,已在很早即開始使用抗菌劑類藥物以進行對於養殖池水、動物養殖、水產養殖等治療動物的疾病及環境之殺菌,這些抗菌劑包括β-內醯胺類抗生素(Beta-lactam)、胺基醣苷類抗生素(Aminoglycoside)、大環內酯類抗生素(Macrolide)、四環素類抗生素(Tetracycline)、氯黴素(Chloramphenicol)、奎諾酮(Quinolone)、磺胺類(Sulfonamides)等藥物;在這些類別藥物中,台灣現今核准使用於水產養殖之動物藥物計有安默西林(Amoxicillin)、安比西林(Ampicillin)、脫氧羥四環黴素(Doxycycline)、紅黴素(Erythromycin)、氟甲磺綠黴素(Florfenicol)、北里黴素(Kitasamycin)、林可黴素(Lincomycin)、歐索林酸(Oxolinic acid)、為歐索林酸衍生物之氟滅菌(Flumequine)、羥四環黴素(Oxytetracycline)、史黴素(Spiramycin)、屬磺胺劑類之磺胺一甲氧嘧啶(Sulfamonomethoxine)、磺胺二甲氧嘧啶(Sulfadimethoxine)、和甲磺綠黴素(Thiamphenicol)等;但,以動物養殖為例,在動物養殖業者用藥知識缺乏下,尤其是未經檢驗確定感染源下,任意連續施用相同類別之藥物造成交叉抗藥性問題,及長期使用下造成病原菌普遍對多數藥物產生抗藥性,致使藥物使用劑量不得不不斷地提高,到最後產生使用藥物無效的結果; 進一步,未遵守停藥期規定而造成藥物殘留的問題,更造成人們食用養殖動物後對健康的威脅。另外,養殖用水任意向外排放,造成包括沿海水域及內陸區域的嚴重污染。 Traditionally, in the application of water treatment in animal husbandry, antibacterial agents have been used very early to carry out sterilization of diseases and environments for therapeutic animals such as pond water, animal breeding, aquaculture, etc. These antibacterial agents include β. - Beta-lactam, Aminoglycoside, Macrolide, Tetracycline, Chloramphenicol, Quinolone ), sulfonamides (Sulfonamides) and other drugs; among these categories of drugs, Taiwan's current approval for aquaculture animal drugs are Amoxicillin, Ampicillin, Doxycycline , Erythromycin, Florfenicol, Kitasamycin, Lincomycin, Oxolinic acid, Oxolinic acid derivatives Flumequine, Oxytetracycline, Spiramycin, Sulfamonomethoxine, Sulfadimeth, Sulfonamide Oxine), and sulfomycin (Thiamphenicol), etc.; however, in the case of animal farming, in the absence of knowledge of animal husbandry medication, especially the unidentified source of infection, any continuous application of the same class of drugs caused crossover The problem of drug resistance, and long-term use, causing pathogens to become generally resistant to most drugs, resulting in a continuous increase in the dose of the drug, and ultimately the result of invalid use of the drug; Further, the problem of drug residues caused by failure to comply with the withdrawal period regulations has caused people to threaten their health after eating animals. In addition, aquaculture water is discharged arbitrarily, causing serious pollution, including coastal waters and inland areas.
以動物養殖為例,採用非上述藥物的化學殺菌劑是另一種選擇,但常用的四級胺對革蘭氏陰性菌殺菌力較差,且對池水中提供溶氧重要來源的藻類具有強烈毒性;醛類諸如甲醛和戊二醛則是致癌物,禁止使用於動物養殖場;氯化合物(諸如次氯酸)的使用會產生三氯甲烷致癌物;碘化合物諸如優碘對魚蝦具刺激性且對部份革蘭氏陰性菌殺菌力不佳。動物養殖場確實需要更安全且更具廣效效能的殺菌劑。 Taking animal breeding as an example, the use of chemical bactericides other than the above drugs is another option, but the commonly used quaternary amines have poor bactericidal power against Gram-negative bacteria and are highly toxic to algae which provide an important source of dissolved oxygen in the pool water; Aldehydes such as formaldehyde and glutaraldehyde are carcinogens and are prohibited for use in animal farms; the use of chlorine compounds such as hypochlorous acid produces chloroform carcinogens; iodine compounds such as iodine are irritating to fish and shrimp. Poor bactericidal power against some Gram-negative bacteria. Animal farms do require a safer and more versatile fungicide.
過醋酸具有的強烈殺菌力、廣效性及生物可分解的環保親和性,早時即使用在醫療器械(其包括塑膠製品)的高層次消毒,也有使用在食品加工廠做為在固體表面、容器和輸送管線的消毒清潔處理。 Peracetic acid has strong bactericidal power, broad-spectrum and biodegradable environmental affinity. It has been used in high-level disinfection of medical devices (including plastic products), and also used in food processing plants as solid surfaces. Disinfection and cleaning of containers and transfer lines.
過醋酸的製備可以醋酸、過氧化氫和水之混合溶液,或以固體諸如四乙醯乙二胺(tetraacetylethylenediamine,TAED)之類醯胺化合物與無機過氧化物在水中反應產生得到。過醋酸的水相/油相參與比例為98:2,為親水性良好的物質,但對疏水性物質滲透力較差,且過高的劑量會對水產養殖動物產生致命毒性,無法以高劑量以克服對油相的滲透(例如使用比利時的Solvay Interox S.A.公司生產之15%過醋酸溶液對屬於海水魚的細鱗鰈浸泡24小時的半數致死濃度LC50為98.7 ppm(過醋酸濃度為14.8 ppm));此外,池水中包括油脂的疏水性物質亦會影響過醋酸的殺菌效能。到目前為止,幾乎沒有相關之研究報告和文獻 及相關商品報導過醋酸在處理水產養殖池池水的殺菌用途。同樣的,亦無相關報導過醋酸在養殖池水、動物養殖等之殺菌用途。 The preparation of peracetic acid can be carried out by reacting a solution of acetic acid, hydrogen peroxide and water, or by reacting a guanamine compound such as tetraacetylethylenediamine (TAED) with an inorganic peroxide in water. The aqueous phase/oil phase of peracetic acid is 98:2, which is a hydrophilic material, but it has poor penetrability to hydrophobic substances, and too high a dose will cause fatal toxicity to aquaculture animals, and cannot be used at high doses. Overcome the penetration of the oil phase (for example, using a 15% peracetic acid solution produced by Solvay Interox SA of Belgium), the LC50 of the semi-lethal concentration of the seawater fish immersed for 24 hours is 98.7 ppm (peracetic acid concentration of 14.8 ppm) In addition, the hydrophobic substances in the pool water, including grease, also affect the bactericidal efficacy of peracetic acid. So far, there are few related research reports and literatures. And related commodities reported the use of acetic acid in the treatment of aquaculture pond water. Similarly, there is no report on the sterilization of acetic acid in culture ponds, animal breeding, etc.
針對此技藝之需要,本發明揭露一種以過醋酸進行抑制動物養殖場之水內及/或環境固體表面上的病原性微生物的方法,該方法包含對養殖場池水及/或環境固體表面投予:(1)經稀釋之過醋酸,使養殖場池水內及/或環境固體表面上之過醋酸濃度為0.0005至4.9 ppm;和(2)濃度為經稀釋之過醋酸的濃度之0.001至60倍之包含至少9個碳原子的碳鏈之界面活性劑;其中經投予後之經稀釋之過醋酸與經稀釋之界面活性劑對養殖場池水及/或環境固體表面的接觸時間至少1分鐘。該包含至少9個碳原子的碳鏈之界面活性劑能增進過醋酸對動物養殖場中疏水性物質之滲透並進而增強過醋酸之抑制病原性微生物的能力。 In response to the needs of the art, the present invention discloses a method for inhibiting pathogenic microorganisms on the surface of water and/or environmental solids of an animal farm using peracetic acid, the method comprising administering to the pond water and/or environmental solid surface of the farm (1) The diluted peracetic acid is such that the concentration of peracetic acid in the pool water and/or on the surface of the environmental solid is 0.0005 to 4.9 ppm; and (2) the concentration is 0.001 to 60 times the concentration of the diluted peracetic acid. A surfactant comprising a carbon chain of at least 9 carbon atoms; wherein the contact between the diluted peracetic acid and the diluted surfactant to the farm pool water and/or the ambient solid surface is at least 1 minute. The surfactant comprising a carbon chain of at least 9 carbon atoms enhances the penetration of peracetic acid into hydrophobic materials in animal farms and thereby enhances the ability of peracetic acid to inhibit pathogenic microorganisms.
本發明發現雖然親水性良好的過醋酸對病原菌有良好抑制能力,但對疏水性物質之滲透性較差,但以經稀釋之過醋酸的濃度之0.001至60倍之包含至少9個碳原子的碳鏈之界面活性劑,可經由提昇過醋酸對疏水物質的滲透力而增強過醋酸的抑菌力。其中過醋酸的來源可由熟知的有機化學反應得到,但不限定為由過氧化氫、醋酸和水的溶液,也可以在酸的催化下以乙醛、乙醯氯、或乙酸酐經氧化作用產生的溶液,也可以為由N-醯胺化合物、過氧化物鹽及水所產生的水溶液得到。 The present inventors have found that although peracetic acid having good hydrophilicity has good inhibitory ability against pathogenic bacteria, it has poor permeability to hydrophobic substances, but contains carbon of at least 9 carbon atoms at a concentration of 0.001 to 60 times the concentration of diluted peracetic acid. The surfactant of the chain enhances the bacteriostasis of peracetic acid by increasing the penetration of peracetic acid into the hydrophobic material. The source of peracetic acid can be obtained by a well-known organic chemical reaction, but is not limited to a solution of hydrogen peroxide, acetic acid and water, or can be produced by oxidation of acetaldehyde, ethyl chloroform or acetic anhydride under acid catalysis. The solution may also be obtained from an aqueous solution produced from an N-guanamine compound, a peroxide salt, and water.
疏水物質包括含有或殘留有飼料油脂的養殖池水、飼料攪拌槽、飼料輸送管、與飼料供料槽、和欄舍 的固體表面等、細胞壁所含具有疏水性胜肽之聚醣的革蘭氏陽性菌和革蘭氏陰性菌、細胞壁含有含氮多醣的幾丁質的真菌、禽畜動物和魚類的皮膚、含有幾丁質的蝦類類外殼、及魚體皮膚潰瘍所產生疏水性物質等,但不限於前所述及的物質和器材設備。 Hydrophobic substances include culture pond water containing or containing feed fats and oils, feed agitation tanks, feed conveying pipes, feed tanks, and barns a solid surface, etc., a Gram-positive and Gram-negative bacteria containing a glycan of a peptide, a cell wall containing a chitin containing a nitrogen-containing polysaccharide, a skin of a poultry animal, and a fish, containing The chitin-like shell of the shrimp and the hydrophobic substance produced by the skin ulcer of the fish, but not limited to the materials and equipment mentioned above.
習知的方法以增加過醋酸濃度是可以提昇過醋酸本身的抑菌力並加速其抑菌速率,但由於過醋酸的毒性會引起禽畜動物呼吸道刺激和皮膚過敏,尤其是極低濃度下的過醋酸也可能會對水產養殖動物產生致命的毒性,且過低濃度的過醋酸往往對某些微生物無法發生有效的抑制作用。本發明發現使用濃度為經稀釋之過醋酸的濃度之0.001至60倍之包含至少9個碳原子的碳鏈之界面活性劑能促進經稀釋之過醋酸之抑菌力。 The conventional method to increase the concentration of peracetic acid can increase the antibacterial activity of peracetic acid itself and accelerate its antibacterial rate, but the toxicity of peracetic acid can cause respiratory irritation and skin irritation in livestock animals, especially at very low concentrations. Peracetic acid may also cause lethal toxicity to aquaculture animals, and too low concentrations of peracetic acid often do not effectively inhibit certain microorganisms. The present inventors have found that the use of a surfactant having a carbon chain of at least 9 carbon atoms at a concentration of from 0.001 to 60 times the concentration of the diluted peracetic acid promotes the bacteriostatic action of the diluted peracetic acid.
以下為以水產養殖場之應用為實施例,但本發明並不限於該實施例,而可應用於養殖池水、動物養殖,其相關實施例之環境以及其應用數據不再贅述。 The following is an example of the application of the aquaculture farm, but the present invention is not limited to the embodiment, but can be applied to culture pond water, animal breeding, the environment of the related embodiments and the application data thereof are not described again.
在本發明之一實施態樣中,使用在禽畜養殖場環境及器具經稀釋之過醋酸濃度為0.0005至18%。 In one embodiment of the invention, the concentration of peracetic acid used in the environment and equipment of the livestock farm is between 0.0005 and 18%.
在本發明之一實施態樣中,過醋酸可以過氧化氫、醋酸、水、在酸的催化下製備的溶液。 In one embodiment of the invention, the peracetic acid can be a solution prepared by the reaction of hydrogen peroxide, acetic acid, water, or acid.
在本發明之一實施態樣中,過醋酸可以過氧化物鹽或過氧化氫在水中與N-醯胺化合物反應製備的溶液。 In one embodiment of the invention, peracetic acid can be a solution prepared by reacting a peroxide salt or hydrogen peroxide with an N-guanamine compound in water.
在本發明之一實施態樣中,過醋酸可以為乙醛、乙醯氯、或乙酸酐經氧化作用產生的溶液。 In one embodiment of the invention, the peracetic acid may be a solution of acetaldehyde, ethyl hydrazine, or acetic anhydride produced by oxidation.
本發明中所使用的濃度計算單位ppm為百萬分之一(part per million)。 The concentration used in the present invention is calculated in ppm per part per million.
在本發明之一實施態樣中,界面活性劑為包含至少9個碳原子的碳鏈之陰離子界面活性劑、包含至少9個 碳原子的碳鏈之非離子界面活性劑、包含至少9個碳原子的碳鏈之兩性離子界面活性劑、或彼等之混合。 In one embodiment of the invention, the surfactant is an anionic surfactant comprising a carbon chain of at least 9 carbon atoms, comprising at least 9 A nonionic surfactant of a carbon chain of a carbon atom, a zwitterionic surfactant comprising a carbon chain of at least 9 carbon atoms, or a mixture thereof.
在本發明之一實施態樣中,該包含至少9個碳原子的碳鏈之陰離子界面活性劑包括:包含至少9個碳原子的碳鏈之烷基硫酸鹽、包含至少9個碳原子的碳鏈之聚氧乙烯基醚硫酸鹽、包含至少9個碳原子的碳鏈之聚氧乙烯烷基苯基醚硫酸鹽、包含至少9個碳原子的碳鏈之烷基磷酸鹽、包含至少9個碳原子的碳鏈之烷基磷酸酯鹽、包含至少9個碳原子的碳鏈之聚氧乙烯烷基苯基醚磷酸鹽、包含至少9個碳原子的碳鏈之脂肪酸鹽、包含至少9個碳原子的碳鏈之烷基磺酸鹽、包含至少9個碳原子的碳鏈之烷基苯磺酸鹽、包含至少9個碳原子的碳鏈之α-烯烴磺酸鹽、包含至少9個碳原子的碳鏈之α-磺化脂肪酸鹽、包含至少9個碳原子的碳鏈之過氧羧酸、包含至少9個碳原子的碳鏈之過氧羧酸之鹼金族鹽、包含至少9個碳原子的碳鏈之羧酸、包含至少9個碳原子的碳鏈之羧酸鹼金族鹽、及彼等之混合。 In one embodiment of the invention, the anionic surfactant comprising a carbon chain of at least 9 carbon atoms comprises: an alkyl sulfate comprising a carbon chain of at least 9 carbon atoms, a carbon comprising at least 9 carbon atoms Chain polyoxyethylene ether sulfate, polyoxyethylene alkylphenyl ether sulfate containing carbon chain of at least 9 carbon atoms, alkyl phosphate containing at least 9 carbon atoms, containing at least 9 An alkyl phosphate salt of a carbon chain of a carbon atom, a polyoxyethylene alkylphenyl ether phosphate of a carbon chain containing at least 9 carbon atoms, a fatty acid salt of a carbon chain containing at least 9 carbon atoms, and at least 9 An alkyl sulfonate of a carbon chain of a carbon atom, an alkylbenzene sulfonate comprising a carbon chain of at least 9 carbon atoms, an α-olefin sulfonate comprising a carbon chain of at least 9 carbon atoms, comprising at least 9 An α-sulfonated fatty acid salt of a carbon chain of a carbon atom, a peroxycarboxylic acid comprising a carbon chain of at least 9 carbon atoms, an alkali metal salt of a peroxycarboxylic acid comprising a carbon chain of at least 9 carbon atoms, comprising at least a carboxylic acid of a carbon chain of 9 carbon atoms, a carboxylic acid alkali gold salt of a carbon chain containing at least 9 carbon atoms And their mixture.
在本發明之一實施態樣中,該包含至少9個碳原子的碳鏈之非離子界面活性劑包括:包含至少9個碳原子的碳鏈之聚氧乙烯烷基醚、包含至少9個碳原子的碳鏈之聚氧乙烯烷基苯基醚、包含至少9個碳原子的碳鏈之聚氧乙烯-聚氧丙烯醇、及彼等之混合。 In one embodiment of the invention, the non-ionic surfactant comprising a carbon chain of at least 9 carbon atoms comprises: a polyoxyethylene alkyl ether comprising a carbon chain of at least 9 carbon atoms, comprising at least 9 carbons A polyoxyethylene alkylphenyl ether of a carbon chain of an atom, a polyoxyethylene-polyoxypropenol containing a carbon chain of at least 9 carbon atoms, and a mixture thereof.
在本發明之一實施態樣中,該包含至少9個碳原子的碳鏈之過氧羧酸包括過氧壬酸(peroxynonanoic acid)、過氧癸酸(peroxydecanoic acid)、過氧十一烷酸(peroxyundecanoic acid)、過氧十二烷酸(peroxydodecanoic acid)或彼等之混合。 In one embodiment of the invention, the peroxycarboxylic acid comprising a carbon chain of at least 9 carbon atoms comprises peroxynonanoic acid, peroxydecanoic acid, peroxyundecanoic acid. (peroxyundecanoic acid), peroxydodecanoic acid or a mixture of them.
在本發明之一實施態樣中,該包含至少9個碳原子 的碳鏈之羧酸鹽包括壬酸(nonanoic acid)、癸酸(decanoic acid)、十一烷酸(undecanoic acid)、月桂酸(dodecanoic acid)、十三烷酸(tridecanoic acid)、肉豆蔻酸(tetradecanoic acid)、前述物質之鹼金族鹽或彼等之混合。 In one embodiment of the invention, the at least 9 carbon atoms are included The carboxylate of the carbon chain includes nonanoic acid, decanoic acid, undecanoic acid, dodecanoic acid, tridecanoic acid, myristic acid. (tetradecanoic acid), an alkali metal salt of the foregoing, or a mixture thereof.
在本發明之一實施態樣中,可使用金屬離子螯合劑以避免水中金屬離子對過醋酸之催化分解,其中該金屬離子螯合劑包括乙烯二胺四乙酸(Ethylenediaminetetraacetic acid,EDTA)及其鈉鹽、二乙基三胺五乙酸(Diethyltriaminepentaacetic acid,DTPA)及其鈉鹽、聚磷酸鹽、有機磷酸鹽、磷酸酯、聚丙烯酸酯、有機磷酸鹽、葡萄酸鈉、及彼等之混合。 In one embodiment of the present invention, a metal ion chelating agent may be used to avoid catalytic decomposition of peracetic acid by metal ions in the water, wherein the metal ion chelating agent comprises Ethylenediaminetetraacetic acid (EDTA) and its sodium salt. Diethyltriaminepentaacetic acid (DTPA) and its sodium salt, polyphosphate, organophosphate, phosphate, polyacrylate, organophosphate, sodium gluconate, and mixtures thereof.
在本發明之一實施態樣中,界面活性劑可包含pH緩衝劑係使經稀釋之過醋酸與經稀釋之界面活性劑在養殖場池水及/或環境固體表面之pH值介於6-11。 In one embodiment of the present invention, the surfactant may comprise a pH buffering agent such that the pH of the diluted peracetic acid and the diluted surfactant on the farm water and/or the environmental solid surface is between 6 and 11 .
在本發明之一實施態樣中,pH緩衝劑包含碳原子數少於9的羧酸,將經稀釋之過醋酸與經稀釋之界面活性劑在養殖場池水及/或環境固體表面之pH值介於6-11。 In one embodiment of the invention, the pH buffer comprises a carboxylic acid having less than 9 carbon atoms and the pH of the diluted peracetic acid and the diluted surfactant on the surface of the farm water and/or environmental solids. Between 6-11.
在本發明之一實施態樣中,疏水性物質包括含有或殘留有飼料油脂的養殖池水、飼料攪拌槽、飼料輸送管、與飼料供料槽、和養殖場欄舍的固體表面等、禽畜動物和魚類的皮膚所含之油脂、蝦類類外殼所含的幾丁質、及魚體皮膚潰瘍所產生疏水性物質等、革蘭氏陽性菌和革蘭氏陰性菌之細胞壁含胜肽聚醣、及真菌微生物之細胞壁的含氮多醣之幾丁質,但不限於前所述及的物質和受污染的器材設備。 In an embodiment of the present invention, the hydrophobic substance includes a culture pond water containing or containing residual feed oil, a feed agitation tank, a feed conveying pipe, a feed supply tank, and a solid surface of a farm fence, and the like. The oil contained in the skin of animals and fish, the chitin contained in the shell of shrimps, and the hydrophobic substances produced by skin ulcers of fish, etc., the cell wall of Gram-positive bacteria and Gram-negative bacteria The chitin of the nitrogen-containing polysaccharide of the cell wall of sugar and fungal microorganisms, but is not limited to the materials mentioned above and the contaminated equipment.
使用過醋酸抑制池水中病原性微生物是一種很好 的方法,過醋酸的水相/油相參與比例為98:2,為親水性良好的物質,但對疏水性物質之滲透性較差。在水產養殖池水中有許多疏水性物質會影響過醋酸之殺菌力,諸如飼料油脂,與治療魚蝦疾病時所使用包覆脂溶性藥物的魚油等,彼等是池水中主要常出現的疏水性物質來源;禽畜養殖場中疏水性物質來自飼料油脂,此會阻礙親水性的過醋酸與池水中微生物接觸,進而減低殺菌效果;當細菌性皮膚疾病發生時,例如親水性產氣單胞菌(Aeromonas hydrophila)、弧菌(Vibrio spp.)、滑走菌(Flexibacter & Cytophaga)常引發石斑魚的細菌性皮膚膿瘍,皮膚與肌肉中的脂肪成份和增生的黏液是過醋酸和魚體接觸的障礙;另外,蝦蟹出現體表外殼症狀時,外殼所含疏水性幾丁質的長鏈多醣也是過醋酸與蝦蟹接觸的障礙;再者,過醋酸對水生養殖動物的毒性特性是提高使用劑量上的限制。 It is a good idea to use peracetic acid to inhibit pathogenic microorganisms in pool water. In the method, the aqueous phase/oil phase of peracetic acid has a ratio of 98:2, which is a hydrophilic substance, but has poor permeability to hydrophobic substances. There are many hydrophobic substances in the aquaculture pond water that affect the bactericidal power of peracetic acid, such as feed fats and oils, and fish oil coated with fat-soluble drugs used in the treatment of fish and shrimp diseases. They are the most common hydrophobicity in pool water. Source of matter; the hydrophobic material in poultry farms comes from feed oils, which prevents hydrophilic peracetic acid from coming into contact with microorganisms in the pool water, thereby reducing the bactericidal effect; when bacterial skin diseases occur, such as hydrophilic Aeromonas (Aeromonas hydrophila), Vibrio spp., and Flexibacter & Cytophaga often cause bacterial skin abscess in grouper, and fat and hyperplastic mucus in skin and muscle are obstacles to contact between acetic acid and fish. In addition, when the shrimp and crab have symptoms of surface shell, the long-chain polysaccharide containing hydrophobic chitin in the outer shell is also a barrier to the contact between peracetic acid and shrimp and crab; in addition, the toxicity characteristic of peracetic acid to aquatic animals is to increase the dosage. limits.
實際上,以海水中數量為2 x 105的溶藻弧菌測試過醋酸的殺菌力時,發現過醋酸濃度從0.015 ppm提高10倍至0.15 ppm都無法在2小時內殺死溶藻弧菌,但過醋酸濃度在0.125 ppm就會造成石斑魚之2吋魚苗出現沉入池底無法上浮現象,經病理解剖可發現鰓部和肝臟嚴重受損,進而迅速死亡。雖然長碳鏈的界面活性劑對魚蝦有強刺激性,但本發明發現使用包含至少9個碳原子的碳鏈之界面活性劑與過醋酸,在適合的浸泡時間下,可安全有效地去除池水中病原性微生物;例如,在2吋半魚苗的海水池中,對稀釋為0.05 ppm過醋酸溶液加入稀釋為0.6 ppm之12倍過醋酸濃度的月桂基硫酸鈉後,可在6小時內將溶藻弧菌全數殺滅,池水在6小時後進行換水,魚體未出現損傷。 In fact, when the bactericidal power of acetic acid was tested in 2 x 10 5 of Vibrio alginolyticus in seawater, it was found that the peracetic acid concentration increased by 10 times from 0.115 ppm to 0.15 ppm, and Vibrio alginolyticus could not be killed within 2 hours. However, if the concentration of peracetic acid is 0.125 ppm, it will cause the 2 carp fry of the grouper to sink into the bottom of the pool and cannot rise. The pathological anatomy can be found that the ankle and liver are severely damaged, and then die rapidly. Although the long carbon chain surfactant is highly irritating to fish and shrimp, the present inventors have found that the use of a surfactant comprising a carbon chain of at least 9 carbon atoms and peracetic acid can be safely and effectively removed at a suitable soaking time. Pathogenic microorganisms in pool water; for example, in a seawater pool of 2,5 and a half fry, after adding a dilution of 0.05 ppm peracetic acid solution to a sodium dilauryl sulfate diluted to a concentration of 0.6 ppm 12 times peracetic acid, it can be used within 6 hours. The Vibrio alginolyticus was completely killed, and the pool water was changed after 6 hours, and the fish body was not damaged.
另外,以細菌結構的微觀角度,水產養殖動物常發生的細菌疾病,來自革蘭氏陽性菌者計有諸如:Clostridium botulinm(肉毒桿菌症)、Renibacterium salmoninarum(細菌性腎臟病;Bacterial Kidney Disease/BKD)、Staphylococcus sp.(葡萄球菌症;眼病)、Streptococcus sp.(鏈球菌症)、Enterococcus/Lactococcus sp.(鏈球菌症)、Lactobacillus sp.(假腎病)、Mycobacterium marinum/Mycobacterium sp.(結核分枝桿菌症)、Nocardia asteroids/Nocardia kampachi(努卡氏菌症)、Staphylococcus epidermidis(表皮葡萄球菌)等;來自革蘭氏陰性菌計有諸如:Flexibacter/Cytophaga/Flavobacterium(鰓病;柱狀病;冷水病)、Edwardsiella ictaluri(河鯰腸道敗血症)、E.tarda(鰻肝腎病),Yersinia ruckeri(紅嘴病)、Citrobacter spp.、Pasteurella piscicida(Photobacterium damsela subsp.piscicida,巴氏桿菌症)、Serratia sp.(敗血症)、Pseudomonas fluorescens(鯉全身性敗血症)、Ps.Anguilliseptica(鰻赤點病)、Vibrio sp.(弧菌症)、Aeromonas salmonicida(癤瘡病)、Aer.hydrophila(鰻赤鰭病;出血性敗血症)、Aer.Sobria/Chlamydiaceae/Rickettsiales(絕對體內寄生;敗血症)等。這些革蘭氏陽性菌和革蘭氏陰性菌之細胞壁中都含疏水性胜肽聚醣成份,其中革蘭氏陽性菌具較厚之細胞壁且含高含量之胜肽聚醣;真菌細胞壁則含有含氮多醣的幾丁質;分枝桿菌和努卡氏菌之細菌壁則含有高達20至30個碳之長鏈具蠟質特性的分枝酸,使其相對於親水性的過醋酸猶如一層疏水性屏障,其中分枝桿菌已在鱉養殖場中出現嚴重感染現象;常見之魚皮膚症 狀係諸如:(1)淡水及淡鹹水魚在12℃至18℃水溫常出現真菌類的水黴菌症Oomycetosis(Achlya and Saprolegnia),會附著在體表出現黏黏的棉絮狀物;(2)屬革蘭氏陰性菌的弧菌造成石斑魚及包括金目鱸、七星鱸和加州鱸等的皮膚白點症狀的菌斑,包含病原菌和黏液等物質,其中石斑魚2吋魚養殖場多採極高密度養殖,並以每2小時排流水比進行換水方式,每一次換水就是一次新的感染的開始,讓弧菌皮膚病變得更棘手;(3)屬革蘭氏陰性菌的非典型產氣單胞菌感染症,出現症狀為皮膚潰瘍症狀。以上的皮膚症狀都有黏質棉絮狀物(水黴菌症)、細菌釋放出蛋白質溶解脢分解肌肉蛋白質所產生的胜肽物質,伴隨油脂與體表增生的黏液等共同皮膚潰瘍特徵,這是屬於水中微生物致使魚隻體表的感染,而魚苗階段餵食赤尾青和豐年蝦幼蟲等活體餌料,甚至是餵食肉食性養殖魚類的下雜魚,都可能帶有病原菌,會造成魚隻自體內向體表的感染症狀。傳統上餵食抗生素和使用抗生素進行藥浴是常採取的方法,但因為長期使用造成多數病菌產生嚴重的抗藥性,即使提高用量也無法殺死病菌,且尤其是浸泡過抗生素的魚苗和蝦苗體質較不良,當放養至室外魚池容易造成適應不良而死亡。在解決這些問題上,在池水中使用益生菌,以優勢數量的有益菌分解有機廢棄物和抑制病原菌的生長是很好的方法,但大量的益生菌反而會成為病毒在水中生存的載體,進而助長病毒感染的機會,使得益生菌的使用成為有益和有害之衝突性做法。過醋酸對於水中養殖魚蝦及藻類的毒性相對其他殺菌劑低得很多,降解後產生無害的醋酸、水和氧氣,適合使用在養殖池的殺菌淨化用途;但池水包括油脂、細胞壁所含具有疏水 性胜肽之聚醣的革蘭氏陽性菌和革蘭氏陰性菌、細胞壁含有含氮多醣的幾丁質的真菌、含有幾丁質的蝦類類外殼、及魚體皮膚潰瘍所產生疏水性物質,都是造成工作濃度最多只有幾個ppm的過醋酸(濃度過高對水中動物會產生毒性反應)無法發揮效能的主要原因。 In addition, bacterial diseases often occur in aquaculture animals from the microscopic point of bacterial structure, such as: Clostridium botulinm (botulinum bacillus), Renibacterium salmoninarum (bacterial kidney disease; Bacterial Kidney Disease/) BKD), Staphylococcus sp. (staphylococcus; eye disease), Streptococcus sp. (streptococcus), Enterococcus/Lactococcus sp. (streptomycin), Lactobacillus sp. (pseudylosis), Mycobacterium marinum/Mycobacterium sp. Mycobacteria), Nocardia asteroids/Nocardia kampachi (Norcaosis), Staphylococcus epidermidis (S. epidermidis), etc.; from Gram-negative bacteria such as: Flexibacter/Cytophaga/Flavobacterium (ricky; columnar disease) Cold water disease), Edwardsiella ictaluri, E. tarda, Yersinia ruckeri, Citrobacter spp., Pasteurella piscicida (Photobacterium damsela subsp. piscicida, Pasteurella) , Serratia sp., sepulmonas fluorescens, Ps. Anguilliseptica . Vibrio sp (Vibrio Disease), Aeromonas salmonicida (furunculosis), Aer.hydrophila (eel red fin disease; hemorrhagic septicemia), Aer.Sobria / Chlamydiaceae / Rickettsiales (absolute endoparasitic; sepsis) and the like. The cell walls of these Gram-positive and Gram-negative bacteria contain hydrophobic peptide glycan glycosides, of which Gram-positive bacteria have thicker cell walls and contain high levels of peptide peptidoglycan; fungal cell walls contain The chitin of the nitrogen-containing polysaccharide; the bacterial wall of the mycobacteria and the nucleus bacteria contains a long chain of waxy properties of up to 20 to 30 carbons, making it a layer relative to the hydrophilic peracetic acid. Hydrophobic barrier, in which mycobacteria have been found to cause serious infections in alfalfa farms; common fish skin disease Such as: (1) freshwater and brackish water fish often appear fungus-like watery mildew Oomycetosis (Achlya and Saprolegnia) at 12 ° C to 18 ° C water temperature, will adhere to the surface of the sticky sticky cotton; (2 ) Vibrio, which is a Gram-negative bacterium, causes groupers and plaques including white-spotted symptoms such as gold mites, seven-star mites and California cockroaches, including pathogens and mucus, among which grouper 2 carp farms are highly dense. Breeding, and changing the water ratio every 2 hours, each time changing water is the beginning of a new infection, making Vibrio skin disease more difficult; (3) Atypical gas producing unit of Gram-negative bacteria Infected with bacteria, the symptoms are symptoms of skin ulcers. The above skin symptoms are characterized by sticky cotton batt (water mold), bacteria releasing protein peptides, protein peptides produced by the decomposition of muscle protein, and common skin ulcer characteristics such as mucus of oil and body surface hyperplasia. Microorganisms in the water cause infection on the surface of the fish, while live feeds such as red-tailed green and brine shrimp larvae are fed at the fry stage, and even the next fish that feed on carnivorous farmed fish may carry pathogenic bacteria, which may cause the fish to be in vivo from the body. Symptoms of infection in the table. Traditionally, feeding antibiotics and using antibiotics for medicated baths is a common method. However, most of the bacteria cause serious drug resistance due to long-term use. Even if the dosage is increased, the bacteria cannot be killed, and especially the fry and shrimps that have been soaked with antibiotics. Poor, when stocked to an outdoor fish pond, it is easy to cause maladaptation and death. In solving these problems, it is a good method to use probiotics in pool water to decompose organic waste with a beneficial amount of beneficial bacteria and inhibit the growth of pathogenic bacteria, but a large number of probiotics will become carriers of virus survival in water, and then The opportunity to promote viral infection makes the use of probiotics a useful and harmful conflicting practice. Peracetic acid is much less toxic to fish, shrimp and algae in water than other fungicides. It produces harmless acetic acid, water and oxygen after degradation. It is suitable for sterilization and purification in culture ponds. However, pool water includes oil and water. Gram-positive and Gram-negative bacteria of the glycopeptide of the peptide, the chitin fungus containing the nitrogen-containing polysaccharide on the cell wall, the shell of the shrimp containing the chitin, and the hydrophobicity of the skin ulcer of the fish Substances are the main cause of the inability to perform peracetic acid (the concentration of which is too toxic to animals in water) at a working concentration of at most a few ppm.
而禽畜養殖動物常發生的細菌疾病,來自革蘭氏陽性菌者計有諸如:Staphylococcus aureus(金黃色葡萄球菌)、Streptococcus faecalis(糞腸球菌)等來自革蘭氏陰性菌計有諸如:Pssuedomonas aeruginosa(綠膿桿菌)、Salmonella typhimurium(鼠傷寒桿菌)、Salmonella choleraesuis(豬霍亂桿菌)、Bacillus cereus(臘桿菌)、E.coli(大腸桿菌)等;來自真菌計有諸如:Candida albicans(白色念珠球菌)、Trichophylton mentagrophytes(鬚瘡小芽胞癬菌)、Aspergillus fumigatus(薰烟色麴菌)、A.flavus(黃麴菌)、Penicillium sp.(盤尼西林菌屬)、Scopulariopsis sp.(小帚樣黴菌屬)等,這些細菌的細胞壁中同樣都含有胜肽聚醣成份,再加上環境表面上的油脂髒污等,都是能影響過醋酸殺菌力的疏水性物質。 Bacterial diseases often occur in poultry-bred animals, such as: Staphylococcus aureus (Staphylococcus aureus), Streptococcus faecalis (E. faecalis), etc. from Gram-negative bacteria such as: Pssuedomonas Aeruginosa, Pseudoella typhimurium, Salmonella choleraesuis, Bacillus cereus, E. coli, etc.; Cocci, Trichophylton mentagrophytes, Aspergillus fumigatus, A. flavus, Penicillium sp., Scopulariopsis sp. Dependent), etc., these bacteria also contain the peptide glycopeptide in the cell wall, and the grease on the surface of the environment is a hydrophobic substance that can affect the bactericidal power of acetic acid.
本發明使用包含至少9個碳原子的碳鏈之界面活性劑以增進對動物養殖場中上述疏水性物質的滲透,進而增強過醋酸的殺菌力以抑制動物養殖場中病原性微生物。 The present invention uses a carbon chain intercalating agent comprising at least 9 carbon atoms to enhance penetration of the above hydrophobic materials in an animal farm, thereby enhancing the bactericidal power of peracetic acid to inhibit pathogenic microorganisms in animal farms.
為更進一步闡述本發明為達成預定發明目的所採取的技術手段及功效,對依據本發明提出的以過醋酸在動物養殖場中抑制病原性微生物的方法及其應用其具體實施方式、特徵及其功效,詳細說明如後。 To further explain the technical means and efficacy of the present invention for achieving the intended purpose of the invention, the method and the application thereof for inhibiting pathogenic microorganisms in animal farms by using peracetic acid according to the present invention, and the specific embodiments and characteristics thereof Efficacy, detailed description as follows.
以下為以水產養殖場之應用為實施例,但本發明並不限於該實施例,而可應用於養殖池水、動物養殖、禽畜養殖,其相關實施例之環境以及其應用數據不再贅述。 The following is an application of the aquaculture farm as an example, but the present invention is not limited to the embodiment, but can be applied to culture pond water, animal breeding, poultry breeding, and the environment of the related embodiments and application data thereof will not be described again.
下述之實施方式僅為進一步說明本發明且非為限定本發明之範圍。在本發明之精神下,任何對於該實施方式的修改仍應係屬本發明之範圍。 The following examples are merely illustrative of the invention and are not intended to limit the scope of the invention. Any modifications to this embodiment are still within the scope of the invention in the spirit of the invention.
以經稀釋在海水中濃度為0.045 ppm之過醋酸做為對照組,另一實驗組為使用同樣經稀釋在海水中濃度為0.045 ppm之過醋酸再加上經稀釋在海水中濃度是過醋酸40倍為1.8 ppm的一種界面活性劑-月桂基硫酸鈉,用以處理長度約8吋半的點帶石斑魚(青斑)皮膚上因弧菌感染所產生的白色菌斑。方法為將病魚分別浸泡在二組海水溶液中,浸泡時間為12小時,同樣方式連續浸泡3次後觀察魚隻皮膚上白色菌斑的變化。結果發現,對照組在三次浸泡過的病魚皮膚上白色菌斑沒有明顯變化;而以另一有添加月桂基硫酸鈉之實驗組浸泡過的病魚皮膚上白色菌斑,於第三次浸泡後白色菌斑消失。 Peracetic acid diluted to a concentration of 0.045 ppm in seawater was used as a control group, and the other experimental group used peracetic acid which was also diluted in seawater at a concentration of 0.045 ppm and diluted in seawater at a concentration of peracetic acid 40. A 1.8 ppm surfactant, sodium lauryl sulfate, is used to treat white plaques from Vibrio infections on the skin of grouper (bleu) with a length of about 8 and a half. The method was to soak the diseased fish in two groups of sea water solution, the soaking time was 12 hours, and the white plaque on the skin of the fish was observed after soaking for 3 times in the same manner. The results showed that the white plaque on the skin of the diseased fish that had been soaked three times did not change significantly in the control group; the white plaque on the skin of the diseased fish that had been soaked in another experimental group with sodium lauryl sulfate was soaked for the third time. After the white plaque disappeared.
在養殖2吋半魚苗的海水池中,海水水溫22.5℃,pH值為8.3,鹽份為35‰,對稀釋為0.05 ppm過醋酸溶液加入稀釋為0.6 ppm之12倍過醋酸濃度的一種界面活性劑-月桂基硫酸鈉後,可在6小時內將溶藻弧菌全數殺滅,池水在6小時後進行換水,魚體未出現損傷。 In the seawater tank where 2 吋 half of the fry are cultured, the sea water temperature is 22.5 ° C, the pH is 8.3, the salt is 35 ‰, and an interface diluted to a dilution of 0.6 ppm of 12 times peracetic acid is added to the 0.05 ppm peracetic acid solution. After the active agent-sodium lauryl sulfate, the Vibrio alginolyticus can be completely killed within 6 hours, and the pool water was changed after 6 hours, and the fish body was not damaged.
製備A、B和C三組包含不同成分之界面活性劑的過醋酸殺菌水溶液,A組含37.7 mg/L的過醋酸、B組 含37.7 mg/L的過醋酸、88 mg/L的十二烷基苯磺酸鈉、及47 mg/L的聚氧乙烯十二烷基醚。C組含18.8 mg/L的過醋酸、16.1 mg/L的過氧癸酸、88 mg/L的十二烷基苯磺酸鈉、及47 mg/L的聚氧乙烯十二烷基醚。在20℃下測試三組殺菌液對數量5x106cfu/ml之綠膿桿菌、金黃色葡萄球菌、及白色念球菌的殺滅率。 Preparation of A, B and C three groups of surfactants containing different components of the peracetic acid sterilizing aqueous solution, group A containing 37.7 mg / L of peracetic acid, group B containing 37.7 mg / L of peracetic acid, 88 mg / L of twelve Sodium alkylbenzene sulfonate and 47 mg/L polyoxyethylene lauryl ether. Group C contained 18.8 mg/L peracetic acid, 16.1 mg/L peroxydecanoic acid, 88 mg/L sodium dodecylbenzenesulfonate, and 47 mg/L polyoxyethylene lauryl ether. The killing rate of three groups of sterilizing liquid against Pseudomonas aeruginosa, Staphylococcus aureus, and Candida albicans in the amount of 5x10 6 cfu/ml was tested at 20 °C.
測試方法:綠膿桿菌和金黃色葡萄球菌的測試依照EN 1276,白色念球菌依照EN 1650的測試。 Test methods: Pseudomonas aeruginosa and Staphylococcus aureus were tested according to EN 1276, and Candida albicans was tested according to EN 1650.
結果如下列的(一)、(二)、(三)表: The results are as follows (1), (2), (3):
由上表(一)、(二)及(三)可發現,含有界面活性劑的 殺菌液,尤其對於屬革蘭氏陰性菌的綠膿桿菌、屬革蘭氏陽性菌的金黃色葡萄球菌和屬真菌的白色念球菌明顯有更強的殺菌力。 It can be found from the above table (1), (2) and (3) that it contains a surfactant. The bactericidal liquid, especially for the gram-negative bacteria Pseudomonas aeruginosa, the gram-positive bacteria Staphylococcus aureus and the fungus-like Candida albicans, have a stronger bactericidal power.
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