2018水質(zhì)總大腸菌群、糞大腸菌群和大腸埃希氏菌的測定酶底物法

目 次前 言 ii1 適用范圍 12 規(guī)范性引用文件 13 術(shù)語和定義 14 方法原理 25 干擾和消除 26 試劑和材料 27 儀器和設(shè)備 38 樣品 39 分析步驟 410 結(jié)果計算與表示 6精密度和準(zhǔn)確度 612 質(zhì)量保證和質(zhì)量控制 813 廢棄物處理 8附錄A（資料性附錄）結(jié)果判定參考圖片 9附錄B（規(guī)范性附錄）97孔定量盤法MPN表 12附錄C（資料性附錄）97孔定量盤法MPN表對應(yīng)95%置信區(qū)間 20iPAGEPAGE1PAGEPAGE2水質(zhì) 總大腸菌群、糞大腸菌群和大腸埃希氏菌的測定 酶底物法1 適用范圍本標(biāo)準(zhǔn)規(guī)定了測定水中總大腸菌群、糞大腸菌群和大腸埃希氏菌的酶底物法。埃希氏菌的測定。本方法的檢出限為10MPN/L。2 規(guī)范性引用文件標(biāo)準(zhǔn)。GB/T6682 分析實驗用水規(guī)格和實驗方法14581 湖泊和水庫采樣技術(shù)指導(dǎo)HJ494水質(zhì)采樣技術(shù)指導(dǎo)HJ/T91地表水和污水監(jiān)測技術(shù)規(guī)范3 術(shù)語和定義下列術(shù)語和定義適用于本標(biāo)準(zhǔn)。3.1總大腸菌群 totalcoliforms37℃培養(yǎng)24h，能產(chǎn)生β-半乳糖苷酶（β-D-galactosidase），分解選擇性培養(yǎng)基中的鄰硝基苯-β-D-吡喃半乳糖苷（ONPG）生成黃色的鄰硝基苯酚的腸桿菌科細(xì)菌。3.2糞大腸菌群 fecalcoliforms又稱耐熱大腸菌群（thermotolerant24h，能產(chǎn)生β-半乳糖苷酶色的鄰硝基苯酚的腸桿菌科細(xì)菌。3.3大腸埃希氏菌 Escherichiacoli俗稱大腸桿菌。37℃培養(yǎng)24培養(yǎng)基中的鄰硝基苯-β-D-吡喃半乳糖苷（ONPG）生成黃色的鄰硝基苯酚，同時產(chǎn)生β-葡萄糖醛酸酶（β-Glucuronidase），分解選擇性培養(yǎng)基中的4-甲基傘形酮-β-D-葡萄糖醛酸苷（MUG）釋放出熒光物質(zhì)（4-甲基傘形酮）的腸桿菌科細(xì)菌。3.4最大可能數(shù) mostprobablenumber（MPN）4 方法原理鄰硝基苯酚（ONP）；大腸埃希氏菌同時又能產(chǎn)生β-葡萄糖醛酸酶，將選擇性培養(yǎng)基中的4-甲基傘形酮-β-D-葡萄糖醛酸苷（MUG）分解為4-甲基傘形酮，在紫外燈照射下產(chǎn)生熒光。統(tǒng)計陽性反應(yīng)出現(xiàn)數(shù)量，查MPN表，分別計算樣品中總大腸菌群、糞大腸菌群、大腸埃希氏菌的濃度值。5 干擾和消除5.1 時加入硫代硫酸鈉溶液（6.4）消除干擾。5.2 重金屬離子具有細(xì)胞毒性，能破壞微生物細(xì)胞內(nèi)的酶活性，導(dǎo)致細(xì)胞死亡，可在樣品采集（8.1）時加入乙二胺四乙酸二鈉溶液（6.5）消除干擾。6 試劑和材料除非另有說明，分析時均使用符合國家標(biāo)準(zhǔn)的分析純試劑，實驗用水應(yīng)滿足GB/T6682中三級水的要求。6.1 培養(yǎng)基。本標(biāo)準(zhǔn)采用MinimalMediumONPG-MUG培養(yǎng)基。每100ml樣品需使用培養(yǎng)基粉末2.7g±0.5g，所含基本成分如下：硫酸銨[(NH4)2SO4] 0.5g硫酸錳（MnSO4） 0.05硫酸鋅（ZnSO4） 0.05硫酸鎂（MgSO4） 10mg氯化鈉（NaCl） 1g氯化鈣（CaCl2） 5mg亞硫酸鈉（Na2SO3） 4mg兩性霉素B（AmphotericinB） 0.1mg鄰硝基苯-β-D-吡喃半乳糖苷（ONPG） 50mg4-甲基傘形酮-β-D-葡萄糖醛酸苷（MUG） 7.5mg茄屬植物萃取物（Solanium萃取物） 50mgN-2-羥乙基哌嗪-N-2-乙磺酸鈉鹽（HEPES鈉鹽） 0.53gN-2-羥乙基哌嗪-N-2-乙磺酸（HEPES） 0.69g也可采用市售商品化培養(yǎng)基制品。6.2 6.3 6.4 硫代硫酸鈉溶液：ρ（Na2S2O3）=0.10g/ml。稱取15.7g硫代硫酸鈉（6.2），溶于適量水中，定容至100ml，臨用現(xiàn)配。6.5 乙二胺四乙酸二鈉溶液：ρ（C10H14N2O8Na2·2H2O）=0.15g/ml。稱取15g乙二胺四乙酸二鈉（6.3），溶于適量水中，定容至100ml，此溶液保質(zhì)期為30d。6.6 97孔定量盤：含49個大孔，48個小孔。其中，每個小孔可容納0.186ml樣品，大孔中48個大孔每個可容納1.86ml樣品，一個頂部大孔可容納11ml樣品。也可采用經(jīng)環(huán)氧乙烷滅菌的市售商品化成品。6.7 標(biāo)準(zhǔn)陽性比色盤。6.8 無菌水：取適量實驗用水，經(jīng)121℃高壓蒸汽滅菌20min，備用。7 儀器和設(shè)備7.1 采樣瓶：具螺旋帽或磨口塞的100ml、250ml、500ml廣口玻璃瓶。注：在采集不存在或不考慮余氧、金屬離子干擾的樣品時，可采用市售無菌采樣瓶或無菌采樣袋。7.2 高壓蒸汽滅菌器：121℃可調(diào)。7.3 恒溫培養(yǎng)箱：允許溫度偏差37℃±1℃、44.5℃±0.5℃。7.4 程控定量封口機：用于97孔定量盤（6.6）的封口。7.5 紫外燈：365～366nm。7.6 移液管：1ml±0.01ml、10ml±0.1ml。也可采用計量合格的可調(diào)式移液器。7.7 三角瓶：100ml。7.8 量筒：100ml±1ml。7.9 一般實驗室常用儀器和設(shè)備。注：三角瓶、移液管、采樣瓶等玻璃器皿及采樣器具要在試驗前按無菌操作要求包扎，121℃高壓蒸汽滅菌20min，烘干，備用。8 樣品8.1 樣品采集點位布設(shè)及采樣頻次按照GB/T14581、HJ/T494和HJ/T91的相關(guān)規(guī)定執(zhí)行。采集微生物樣品時，采樣瓶（7.1）不得用樣品洗滌，采集樣品于滅菌的采樣瓶中。面10～15cm處，瓶口朝水流方向，拔瓶塞，使樣品灌入瓶內(nèi)然后蓋上瓶塞，將采樣瓶從水品采集完畢后，迅速扎上無菌包裝紙。3～5min，然后將龍頭關(guān)閉，用火焰灼燒約3min滅菌或用70%～75%的酒精對龍頭進(jìn)行消毒，開足龍頭，再放水1min，以充分除去水管中的滯留雜質(zhì)。采樣時控制水流速度，小心接入瓶內(nèi)。采集地表水、廢水樣品及一定深度的樣品時，也可使用滅菌過的專用采樣裝置采樣。在同一采樣點進(jìn)行分層采樣時，應(yīng)自上而下進(jìn)行，以免不同層次的攪擾。去活性氯對細(xì)菌的抑制作用（每125ml容積加入0.1ml干擾（每125ml容積加入0.3ml注：15.7mg硫代硫酸鈉（6.2）可去除樣品中1.5mg活性氯，硫代硫酸鈉用量可根據(jù)樣品實際活性氯量調(diào)整。8.2樣品保存采樣后應(yīng)在2h內(nèi)檢測，否則，應(yīng)10℃以下冷藏但不得超過6h。實驗室接樣后，不能立即開展檢測的，將樣品于4℃以下冷藏并在2h內(nèi)檢測。9分析步驟9.1樣品稀釋根據(jù)樣品污染程度確定接種量（見表1），避免接種樣品培養(yǎng)后97孔定量盤（6.6）出現(xiàn)全部陽性或全部陰性。接種量小于100ml時，應(yīng)稀釋樣品后接種，接種量為10ml時，取10ml樣品加入到盛有90ml1:10的稀釋樣品，其他接種量的稀釋樣品依次類推。對于未知樣品，可選用多個接種量進(jìn)行檢測。表1 樣品接種量參考表樣品類型接種量（ml）1021010.110-2地表水水源水▲湖泊（水庫）▲河流▲▲▲廢水生活污水▲▲▲▲工業(yè)廢水處理前▲▲▲▲處理后▲地下水▲9.2接種量取100ml樣品或稀釋樣品于滅菌后的三角瓶，加入2.7g±0.5g培養(yǎng)基（6.1）粉末，充分混勻，完全溶解后，全部倒入97孔定量盤（6.6）內(nèi)，以手撫平97孔定量盤背面，趕除孔內(nèi)氣泡，然后用程控定量封口機（7.4）封口。觀察97孔定量盤顏色，若出現(xiàn)類似或深于標(biāo)準(zhǔn)陽性比色盤（6.7）的顏色，則需排查樣品、培養(yǎng)基、無菌水等一系列因素后，終止試驗或重新操作。注：9.1、9.2步驟在野外操作時應(yīng)避開明顯局部污染源，建議使用一次性手套、口罩、酒精燈等。9.3培養(yǎng)測定總大腸菌群和大腸埃希氏菌時，將封口后的97孔定量盤放入恒溫培養(yǎng)箱（7.3）中37℃±1℃下培養(yǎng)24h。97孔定量盤放入的恒溫培養(yǎng)箱中44.5℃±0.5℃下培養(yǎng)24h。9.4對照試驗9.4.1空白對照每次試驗都要用無菌水（6.8）按照步驟9.1～9.3進(jìn)行實驗室空白測定。培養(yǎng)后的97孔定量盤不得有任何顏色反應(yīng)，否則，該次樣品測定結(jié)果無效，應(yīng)查明原因后重新測定。9.4.2陰性和陽性對照總大腸菌群、大腸埃希氏菌、糞大腸菌群的陰性、陽性菌株參考表2。表2 陰性、陽性菌株參考表檢測指標(biāo)陽性菌種陰性菌種總大腸菌群大腸埃希氏菌、產(chǎn)氣腸桿菌（Enterobacteraerogenes）假單胞菌屬（Pseudomonassp.）大腸埃希氏菌大腸埃希氏菌金黃色葡萄球菌、假單胞菌屬糞大腸菌群產(chǎn)氣腸桿菌、糞鏈球菌（Streptococcus假單胞菌屬將標(biāo)準(zhǔn)菌株制成300～3000個/ml操作，陽性菌株應(yīng)呈現(xiàn)陽性反應(yīng)；陰性菌株呈現(xiàn)陰性反應(yīng)，否則，該次樣品測定結(jié)果無效，應(yīng)重新測定。況用無菌水（6.8）稀釋至300～3000個/ml。9.4.3 結(jié)果判讀與計數(shù)將培養(yǎng)24h后的97孔定量盤進(jìn)行結(jié)果判讀，樣品變黃色判斷為總大腸菌群或糞大腸菌群陽性；樣品變黃色且在紫外燈（7.5）照射下有藍(lán)色熒光，判斷為大腸埃希氏菌陽性。如果結(jié)果可疑，可延長培養(yǎng)至28h進(jìn)行結(jié)果判讀，超過28h后出現(xiàn)的顏色反應(yīng)不作為陽性結(jié)果?？墒褂帽Ｙ|(zhì)期內(nèi)的標(biāo)準(zhǔn)陽性比色盤以輔助判讀，參見附錄A。分別記錄97孔定量盤中大孔和小孔的陽性孔數(shù)量。10 結(jié)果計算與表示10.1 結(jié)果計算從97孔定量盤法MPN表（見附錄B）中查得每100ml樣品中總大腸菌群、糞大腸菌群數(shù)或大腸埃希氏菌的MPN值（置信區(qū)間參見附錄C）后，再根據(jù)樣品不同的稀釋度，按照公式（1）換算樣品中總大腸菌群、糞大腸菌群數(shù)或大腸埃希氏菌濃度（MPN/L）：CMPN值×1000f

（1）式中：C——樣品中總大腸菌群、糞大腸菌群數(shù)或大腸埃希氏菌濃度，MPN/L；MPN值——每100ml樣品中總大腸菌群、糞大腸菌群數(shù)或大腸埃希氏菌濃度，MPN/100ml；1000——將C單位由MPN/ml轉(zhuǎn)換為MPN/L；f——最大接種量，ml。10.2 結(jié)果表示測定結(jié)果保留兩位有效數(shù)字，當(dāng)測定結(jié)果≥100MPN/L時，以科學(xué)計數(shù)法表示；若97孔均為陰性，可報告為總大腸菌群、糞大腸菌群數(shù)或大腸埃希氏菌未檢出或＜10MPN/L。11 精密度和準(zhǔn)確度11.1 精密度6個實驗室分別對低濃度（地下水，濃度均值為6.0×102MPN/L）、中濃度（地表水，濃度均值為4.0×104MPN/L）和高濃度（生活污水，濃度均值為9.0×107MPN/L）三個不同濃度細(xì)菌總數(shù)的樣品及有證標(biāo)準(zhǔn)樣品（濃度為2000MPN/L）進(jìn)行了6次重復(fù)測定：實驗室內(nèi)相對標(biāo)準(zhǔn)偏差范圍分別為0.26%～1.3%、0.65%～2.0%、1.3%～3.7%和0.68%～2.8%；實驗室間相對標(biāo)準(zhǔn)偏差分別為和0.18；再現(xiàn)性限為1.8、0.23、0.38和0.20；實驗室間置信區(qū)間見表3。表3 總大腸菌群實驗室間95%置信區(qū)間表低濃度（MPN/L）中濃度（MPN/L）高濃度（MPN/L）均值95%置信區(qū)間均值95%置信區(qū)間均值95%置信區(qū)間均值95%置信區(qū)間6.4×1025.5×102～7.4×1024.0×1043.5×104～4.6×1044.1×1063.7×106～4.6×1061.4×1031.8×103～2.3×1036個實驗室分別對低濃度（地下水，濃度均值為70MPN/L）、中濃度（地表水，濃度均值為9.0×103MPN/L）和高濃度（生活污水，濃度均值為2.0×106MPN/L）三個不同濃度細(xì)菌總數(shù)的樣品及有證標(biāo)準(zhǔn)樣品（濃度為2000MPN/L）進(jìn)行了6次重復(fù)測定：實驗室內(nèi)相對標(biāo)準(zhǔn)偏差范圍分別為0.69%～1.9%、0.93%～2.9%、5.4%～21%和1.8%～3.0%；實驗室間相對標(biāo)準(zhǔn)偏差分別為9.6%、1.6%、15%和3.7%；重復(fù)性限為0.23、0.21、0.58和0.19；再現(xiàn)性限為1.7、0.26、0.90和0.35；實驗室間置信區(qū)間見表4。表4 大腸埃希氏菌實驗室間置信區(qū)間表低濃度（MPN/L）中濃度（MPN/L）高濃度（MPN/L）有證標(biāo)準(zhǔn)樣品（MPN/L）均值95%置信區(qū)間均值95%置信區(qū)間均值95%置信區(qū)間均值95%置信區(qū)間5939～908.5×1037.3×103～9.9×1031.5×1061.3×106～1.7×1067.7×1021.7×103～2.3×1036個實驗室分別對低濃度（地下水，濃度均值為1.0×102MPN/L）、中濃度（地表水，濃度均值為1.3×104MPN/L）和高濃度（生活污水，濃度均值為3.0×106MPN/L）三個不同濃度細(xì)菌總數(shù)的樣品及有證標(biāo)準(zhǔn)樣品（濃度為3700MPN/L）進(jìn)行了6次重復(fù)測定：實驗室內(nèi)相對標(biāo)準(zhǔn)偏差范圍分別為0.40%～2.4%、1.3%～2.9%、3.4%～17%和2.4%～9.2%；實驗室間相對標(biāo)準(zhǔn)偏差分別為8.9%、6.4%、6.6%、6.4%；重復(fù)性限為0.22、0.26、0.58和0.55；再現(xiàn)性限為1.6、0.76、0.64和0.75；實驗室間置信區(qū)間見表5。表5 糞大腸菌群實驗室間置信區(qū)間表低濃度（MPN/L）中濃度（MPN/L）高濃度（MPN/L）有證標(biāo)準(zhǔn)樣品（MPN/L）均值95%置信區(qū)間均值95%置信區(qū)間均值95%置信區(qū)間均值95%置信區(qū)間9262～1.3×1021.1×1049.5×103～1.4×1041.8×1061.6×106～2.1×1061.4×1032.5×103～5.4×10311.2 準(zhǔn)確度6家實驗室對總大腸菌群有證標(biāo)準(zhǔn)樣品（20006的相對誤差范圍為-6.4%～-3.5%；相對誤差的最終值為-4.7%±1.2%。6家實驗室對大腸埃希氏菌群標(biāo)準(zhǔn)樣品（20006的相對誤差范圍為-15%～-6.3%；相對誤差的最終值為-13%±3.3%。6家實驗室對糞大腸菌群的有證標(biāo)準(zhǔn)樣品（37006內(nèi)的相對誤差范圍為-17%～-0.81%；相對誤差的最終值為-13%±6.1%。注：微生物檢測數(shù)據(jù)為偏態(tài)分布，其測定結(jié)果全部經(jīng)以10為底對數(shù)轉(zhuǎn)換后進(jìn)行計算。12 質(zhì)量保證和質(zhì)量控制12.1 每批樣品按對照試驗（9.4）進(jìn)行空白對照測定，定期使用有證標(biāo)準(zhǔn)菌株進(jìn)行陽性和陰性對照試驗。12.2 每20個樣品或每批次樣品（≤20個/批）測定一個平行雙樣。12.3 對每批次培養(yǎng)基須使用有證標(biāo)準(zhǔn)菌株進(jìn)行培養(yǎng)基質(zhì)量檢驗。12.4 定期使用有證標(biāo)準(zhǔn)菌株/標(biāo)準(zhǔn)樣品進(jìn)行質(zhì)量控制。13 廢物處理實驗中產(chǎn)生的廢物經(jīng)121℃高壓蒸汽滅菌20min后，作為一般廢物處理。附錄A（資料性附錄）結(jié)果判定參考圖片12圖A.1 總大腸菌群、糞大腸菌群酶底物法陰性、陽性結(jié)果參考圖PAGEPAGE13PAGEPAGE1012圖A.2 大腸埃希氏菌酶底物法陰性、陽性結(jié)果參考圖圖A.3 標(biāo)準(zhǔn)陽性比色盤附錄B（規(guī)范性附錄）97孔定量盤法MPN表表B.1 97孔定量盤法MPN表（MPN/100ml）大孔陽性格數(shù)小孔陽性格數(shù)0123456789100<1.01.02.03.04.05.06.07.08.09.010.011.011.02.03.04.05.06.07.18.19.110.111.112.122.03.04.15.16.17.18.19.210.211.212.213.333.14.15.16.17.28.29.210.311.312.413.414.544.15.26.27.28.39.310.411.412.513.514.615.655.26.37.38.49.410.511.512.613.714.715.816.966.37.48.49.510.611.612.713.814.916.017.018.177.58.59.610.711.812.813.915.016.117.218.319.488.69.710.811.913.014.115.216.317.418.519.620.799.810.912.013.114.215.316.417.618.719.820.922.01011.012.113.214.415.516.617.718.920.021.122.323.412.213.414.515.616.817.919.120.221.422.523.724.81213.514.615.816.918.119.320.421.622.823.925.126.31314.816.017.118.319.520.621.823.024.225.426.627.81416.117.318.519.720.922.123.324.525.726.928.129.31517.518.719.921.122.323.524.725.927.228.429.630.91618.920.121.322.623.825.026.227.528.730.031.232.51720.321.622.824.125.326.627.829.130.331.632.934.11821.823.124.325.626.928.129.430.732.033.334.635.91923.324.625.927.228.529.831.132.433.735.036.337.62024.926.227.528.830.131.532.834.135.436.838.139.52126.527.929.230.531.833.234.535.937.338.640.041.42228.229.530.932.333.635.036.437.739.140.541.943.3續(xù)表續(xù)表續(xù)表續(xù)表大孔陽性格數(shù)小孔陽性格數(shù)0123456789102329.931.332.734.135.536.838.339.741.142.543.945.42431.733.134.535.937.338.840.241.743.144.646.047.52533.635.036.437.939.340.842.243.745.246.748.249.72635.536.938.439.941.442.844.345.947.448.950.452.02737.438.940.442.043.545.046.548.149.651.252.854.42839.541.042.644.145.747.348.850.452.053.655.256.92941.743.244.846.448.049.651.252.854.556.157.859.53043.945.547.148.750.452.053.755.457.158.860.562.23146.247.949.551.252.954.656.358.159.861.663.365.13248.750.452.153.855.657.359.160.962.764.566.368.23351.253.054.856.558.360.262.063.865.767.669.571.43453.955.757.659.461.363.165.067.068.970.872.874.83556.858.660.562.464.466.368.370.372.374.376.378.43659.861.763.765.767.769.771.773.875.978.080.182.33762.965.067.069.171.273.375.477.679.882.084.286.53866.368.470.672.774.977.179.481.683.986.288.691.03970.072.274.476.778.981.383.686.088.490.993.495.94073.876.278.580.983.385.788.290.893.395.998.5101.24178.080.583.085.588.090.693.395.998.7101.4104.3107.14282.685.287.890.593.296.098.8101.7104.6107.6110.6113.74387.690.493.296.099.0101.9105.0108.1114.5117.8121.14493.196.199.1102.2105.4108.6115.3118.7122.3125.9129.64599.3102.5105.8109.2112.6116.2119.8123.6127.4131.4135.4139.646106.3109.8113.4117.2121.0125.0129.1133.3137.6142.1146.7151.547114.3118.3122.4126.6130.9135.4140.1145.0150.0155.3160.7166.448123.9128.4133.1137.9143.0148.3153.9159.7165.8172.2178.9186.049135.5140.8146.4152.3158.5165.0172.0179.3187.2195.6204.6214.3大孔陽性格數(shù)小孔陽性格數(shù)121314151617181920212223012.013.014.115.116.117.118.119.120.221.222.223.3113.214.215.216.217.318.319.320.421.422.423.524.5214.315.416.417.418.519.520.621.622.723.724.825.8315.516.517.618.619.720.821.822.923.925.026.127.1416.717.818.819.921.022.023.124.225.326.327.428.5517.919.020.121.222.223.324.425.526.627.728.829.9619.220.321.422.523.624.725.826.928.029.130.231.3720.521.622.723.824.926.027.128.329.430.531.632.8821.822.924.125.226.327.428.629.730.832.033.134.3923.224.325.426.627.728.930.031.232.333.534.635.81024.625.726.928.029.230.331.532.733.835.036.237.426.027.228.329.530.731.933.034.235.436.637.839.01227.528.629.831.032.233.434.635.837.038.239.540.71329.030.231.432.633.835.036.237.538.739.941.242.41430.531.733.034.235.436.737.939.140.441.642.944.21532.133.334.635.837.138.439.640.942.243.444.746.01633.735.036.337.538.840.141.442.744.045.346.647.91735.436.738.039.340.641.943.244.545.947.248.549.81837.238.539.841.142.443.845.146.547.849.250.551.91939.040.341.643.044.345.747.148.449.851.252.654.02040.842.243.644.946.347.749.150.551.953.354.756.12142.844.145.546.948.449.851.252.654.155.556.958.42244.846.247.649.050.551.953.454.856.357.859.360.82346.848.349.751.252.754.255.657.158.660.261.763.22449.050.552.053.555.056.558.059.561.162.664.265.8大孔陽性格數(shù)小孔陽性格數(shù)1213141516171819202122232551.252.754.355.857.358.960.562.063.665.266.868.42653.555.156.758.259.861.463.064.766.367.969.671.22756.057.659.260.862.464.165.767.469.170.872.574.22858.560.261.863.565.266.968.670.372.073.775.577.32961.262.964.666.368.069.871.573.375.176.978.780.53064.065.767.569.371.072.974.776.578.380.282.184.03166.968.770.572.474.276.178.079.981.883.785.787.63270.071.973.875.777.679.581.583.585.487.589.591.53373.375.277.279.281.283.285.287.389.391.493.695.73476.878.880.882.985.087.189.291.493.595.797.9100.23580.582.684.786.989.191.393.595.798.0100.3102.6105.03684.586.788.991.293.595.898.1100.5102.9105.3107.7110.23788.891.193.495.898.2100.6103.1105.6108.1110.7113.3115.93893.495.898.3100.8103.4105.9108.6113.9116.6119.4122.23998.4101.0103.6106.3109.0114.6117.4120.3123.2126.1129.240103.9106.7109.5112.4115.3118.2121.2124.3127.4130.5133.7137.041110.0113.0116.0119.1122.2125.4128.7132.0135.4138.8142.3145.942116.9120.1123.4126.7130.1133.6137.2140.8144.5148.3152.2156.143124.6128.1131.7135.4139.1143.0147.0151.0155.2159.4163.8168.244133.4137.4141.4145.5149.7154.1158.5163.1167.9172.7177.7182.945143.9148.3152.9157.6162.4167.4172.6178.0183.5189.2195.1201.246156.5161.6167.0172.5178.2184.2190.4196.8203.5210.5217.8225.447172.3178.5185.0191.8198.9206.4214.2222.4231.0240.0249.5259.548193.5201.4209.8218.7228.2238.2248.9260.3272.3285.1298.7313.049224.7235.9248.1261.3275.5290.9307.6325.5344.8365.4387.3410.6大孔陽性格數(shù)小孔陽性格數(shù)242526272829303132333435024.325.326.427.428.429.530.531.532.633.634.735.7125.626.627.728.729.830.831.932.934.035.036.137.2226.927.929.030.031.132.233.234.335.436.537.538.6328.229.330.431.432.533.634.735.836.837.939.040.1429.630.731.832.833.935.036.137.238.339.440.541.6531.032.133.234.335.436.537.638.739.941.042.143.2632.433.534.735.836.938.039.240.341.442.643.744.8733.935.036.237.338.439.640.741.943.044.245.346.5835.436.637.738.940.041.242.343.544.745.947.048.2937.038.139.340.541.642.844.045.246.447.648.850.0.1038.639.740.942.143.344.545.746.948.149.350.651.840.241.442.643.845.046.347.548.749.951.252.453.71241.943.144.345.646.848.149.350.651.853.154.355.61343.644.946.147.448.649.951.252.553.755.056.357.61445.446.748.049.350.551.853.154.455.757.058.359.61547.348.649.951.252.553.855.156.457.859.160.461.81649.250.551.853.254.555.857.258.559.961.262.664.01751.252.553.955.256.658.059.360.762.163.564.966.31853.254.656.057.458.860.261.663.064.465.867.268.61955.456.858.259.661.062.463.965.366.868.269.771.12057.659.060.461.963.364.866.367.769.270.772.273.72159.961.362.864.365.867.368.870.371.873.374.976.42262.363.865.366.868.369.871.472.974.576.177.679.22364.766.367.869.471.072.574.175.777.378.980.582.22467.368.970.572.173.775.377.078.680.381.983.685.2大孔陽性格數(shù)小孔陽性格數(shù)2425262728293031323334352570.071.773.375.076.678.380.081.783.385.186.888.52672.974.676.378.079.781.483.184.886.688.490.191.92775.977.679.481.182.984.686.488.290.091.993.795.52879.080.882.684.486.388.189.991.893.795.697.599.42982.484.286.187.989.891.793.795.697.599.5101.5103.53085.987.889.791.793.695.697.699.6101.6103.7105.7107.83189.691.693.695.697.799.7101.8103.9106.0108.2110.3112.53293.695.797.899.9102.0104.2106.3108.5110.7113.0115.2117.53397.8100.0102.2104.4106.6108.9113.5115.8118.2120.5122.934102.4104.7107.0109.3114.0116.4118.9121.3123.8126.3128.835107.3109.7112.2114.6117.1119.6122.2124.7127.3129.9132.6135.336112.7115.2117.8120.4123.0125.7128.4131.1133.9136.7139.5142.437118.6121.3124.0126.8129.6132.4135.3138.2141.2144.2147.3150.338125.0127.9130.8133.8136.8139.9143.0146.2149.4152.6155.9159.239132.2135.3138.5141.7145.0148.3151.7155.1158.6162.1165.7169.440140.3143.7147.1150.6154.2157.8161.5165.3169.1173.0177.0181.141149.5153.2157.0160.9164.8168.9173.0177.2181.5185.8190.3194.842160.2164.3168.6172.9177.3181.9186.5191.3196.1201.1206.2211.443172.8177.5182.3187.3192.4197.6202.9208.4214.0219.8225.8231.844188.2193.6199.3205.1211.0217.2223.5230.0236.7243.6250.8258.145207.5214.1220.9227.9235.2242.7250.4258.4266.7275.3284.1293.346233.3241.5250.0258.9268.2277.8287.8298.1308.8319.9331.4343.347270.0280.9292.4304.4316.9330.0343.6357.8372.5387.7403.4419.848328.2344.1360.9378.4396.8416.0436.0456.9478.6501.2524.7549.349435.2461.1488.4517.2547.5579.4613.1648.8686.7727.0770.1816.4大孔陽性格數(shù)小孔陽性格數(shù)36373839404142434445464748036.837.838.940.041.042.143.144.245.346.347.448.549.5138.239.340.441.442.543.644.745.746.847.949.050.151.2239.740.841.943.044.045.146.247.348.449.550.651.752.8341.242.343.444.545.646.747.848.950.051.252.353.454.5442.843.945.046.147.248.349.550.651.752.954.055.156.3544.445.546.647.748.950.051.252.353.554.655.856.958.1646.047.148.349.450.651.752.954.155.256.457.658.759.9747.748.850.051.252.353.554.755.957.158.359.460.661.8849.450.651.853.054.155.356.557.759.060.261.462.663.8951.252.453.654.856.057.258.459.760.962.163.464.665.81053.054.255.556.757.959.260.461.762.964.265.466.767.954.956.157.458.659.961.262.463.765.066.367.568.870.11256.858.159.460.762.063.264.565.867.168.469.771.072.41358.960.261.562.864.165.466.768.069.370.772.073.374.71460.962.363.664.966.367.668.970.371.673.074.475.777.11563.164.565.867.268.569.971.372.674.075.476.878.279.61665.366.768.169.570.972.373.775.176.577.979.380.882.21767.769.170.571.973.374.876.277.679.180.582.083.584.91870.171.573.074.475.977.378.880.381.883.384.886.387.81972.674.175.577.078.580.081.583.184.686.187.689.290.72075.276.778.279.881.382.884.485.987.589.190.792.293.82177.979.581.182.684.285.887.489.090.692.293.895.497.12280.882.484.085.687.288.990.592.193.895.597.198.8100.52383.885.487.188.790.492.193.895.597.298.9100.6102.4104.12486.988.690.392.093.895.597.299.0100.7102.5104.3106.1107.9大孔陽性格數(shù)小孔陽性格數(shù)363738394041424344454647482590.292.093.795.597.399.1100.9102.7104.5106.3108.2110.02693.795.597.399.2101.0102.9104.7106.6108.5110.4112.3114.2116.22797.499.3101.2103.1105.0106.9108.8110.8112.7114.7116.7118.7120.728101.3103.3105.2107.2109.2113.2115.2117.3119.3121.4123.5125.629105.5107.5109.5113.7115.7117.8120.0122.1124.2126.4128.6130.830109.9112.0114.2116.3118.5120.6122.8125.1127.3129.5131.8134.1136.431114.7116.9119.1121.4123.6125.9128.2130.5132.9135.3137.7140.1142.532119.8122.1124.5126.8129.2131.6134.0136.5139.0141.5144.0146.6149.133125.4127.8130.3132.8135.3137.8140.4143.0145.6148.3150.9153.7156.434131.4134.0136.6139.2141.9144.6147.4150.1152.9155.7158.6161.5164.435138.0140.8143.6146.4149.2152.1155.0158.0161.0164.0167.1170.2173.336145.3148.3151.3154.3157.3160.5163.6166.8170.0173.3176.6179.9183.337153.5156.7159.9163.1166.5169.8173.2176.7180.2183.7187.3191.0194.738162.6166.1169.6173.2176.8180.4184.2188.0191.8195.7199.7203.7207.739173.1176.9180.7184.7188.7192.7196.8201.0205.3209.6214.0218.5223.040185.2189.4193.7198.1202.5207.1211.7216.4221.1226.0231.0236.0241.141199.5204.2209.1214.0219.1224.2229.4234.8240.2245.8251.5257.2263.142216.7222.2227.7233.4239.2245.2251.3257.5263.8270.3276.9283.6290.543238.1244.5251.0257.7264.6271.7278.9286.3293.8301.5309.4317.4325.744265.6273.3281.2289.4297.8306.3315.1324.1333.3342.8352.4362.3372.445302.6312.3322.3332.5343.0353.8364.9376.2387.9399.8412.0424.5437.446355.5368.1381.1394.5408.3422.5437.1452.0467.4483.3499.6516.3533.547436.6454.1472.1490.7509.9529.8550.4571.7593.8616.7640.5665.3691.048574.8601.5629.4658.6689.3721.5755.6791.5829.7870.4913.9960.61011.249866.4920.8980.41046.21203.31299.71413.61553.11732.91986.32419.6＞2419.6PAGEPAGE21PAGEPAGE20附錄C（資料性附錄）97孔定量盤法MPN表對應(yīng)95%置信區(qū)間表C.1 97孔定量盤法MPN表對應(yīng)95%置信區(qū)間（MPN/100ml）大孔陽性格數(shù)小孔陽性格數(shù)012345600.0～3.70.0～3.70.3～5.60.6～7.31.1～8.91.7～10.52.3～12.110.1～5.50.3～5.90.6～7.31.1～8.91.7～10.52.3～12.13.0～13.720.3～7.10.7～7.41.2～9.01.6～10.62.3～12.13.0～13.73.7～15.330.7～8.91.2～9.11.7～10.62.3～12.13.0～13.73.6～15.34.4～16.941.7～9.51.8～10.82.4～12.22.9～13.73.6～15.34.5～16.95.2～18.552.3～11.92.5～12.72.9～13.93.7～15.34.3～16.95.2～18.56.0～20.162.9～13.73.2～14.43.7～15.34.4～16.95.3～18.56.0～20.16.8～21.773.6～14.93.9～15.64.4～16.95.1～18.56.1～20.16.9～21.77.8～22.984.5～16.94.5～17.25.2～18.65.9～20.26.7～21.87.8～23.48.7～24.694.7～18.45.6～19.56.0～20.36.8～21.87.6～23.48.5～25.19.5～26.4105.7～20.16.5～21.17.1～22.07.7～23.68.6～25.19.6～26.810.6～27.56.8～21.47.4～22.37.8～23.68.7～25.39.7～26.810.7～28.211.4～29.3127.8～23.48.2～24.68.8～25.79.4～27.010.4～28.611.5～30.012.6～31.2138.5～25.19.2～26.49.9～27.410.5～28.811.6～30.412.7～31.813.4～33.1149.3～26.810.3～28.211.0～29.211.7～30.712.4～32.313.6～33.714.8～35.01510.1～28.610.8～30.011.9～30.912.6～32.613.7～34.114.9～35.215.7～36.51611.3～30.412.4～31.812.7～32.613.9～34.514.6～35.815.9～37.117.2～38.51712.1～32.212.9～33.714.0～35.014.8～36.516.1～37.716.9～39.218.2～40.61813.4～33.914.2～35.715.4～37.115.7～38.417.1～39.818.4～41.319.2～42.71914.4～36.115.1～37.616.4～39.117.3～40.218.1～41.819.5～43.420.9～44.92015.8～38.116.6～39.717.5～41.218.3～42.719.7～44.220.6～45.722.1～47.22116.8～40.118.2～41.318.5～43.119.4～44.820.8～46.421.7～47.923.3～49.52217.9～42.318.8～44.019.6～45.521.2～47.222.0～48.823.6～50.324.5～51.3續(xù)表續(xù)表續(xù)表續(xù)表大孔陽性格數(shù)小孔陽性格數(shù)01234562319.0～44.420.5～46.021.4～47.722.3～49.223.9～51.024.8～52.225.8～53.82420.7～46.621.7～48.123.3～50.124.2～51.925.2～53.326.1～54.727.1～56.42522.0～48.922.9～51.223.8～52.625.5～54.026.5～55.928.3～57.329.3～59.02623.2～52.024.9～53.725.9～55.426.9～56.527.9～58.629.7～60.130.7～61.82724.5～54.526.2～55.927.3～57.428.3～59.729.3～60.831.2～63.032.3～64.72825.9～57.027.7～58.828.7～60.730.6～62.530.8～63.832.8～65.933.9～67.52928.1～59.829.1～61.430.2～63.431.3～65.433.3～67.234.4～68.535.5～70.73029.6～62.630.7～64.531.8～66.633.8～68.034.9～70.136.1～72.238.3～74.03131.1～65.532.3～67.534.4～69.335.5～71.736.7～73.737.9～75.639.1～77.63232.8～69.033.9～70.936.1～73.037.3～75.038.5～77.239.7～79.142.1～81.23334.5～72.336.7～74.338.0～76.439.2～78.640.5～80.642.9～83.044.2～85.13437.4～75.838.6～77.939.9～80.041.2～82.243.7～84.645.0～86.946.4～89.23539.4～79.540.7～81.542.0～83.743.3～86.244.6～88.647.3～90.448.7～93.03641.4～83.442.8～85.444.1～87.845.5～90.346.9～92.749.7～95.351.1～97.53743.6～87.445.0～89.646.5～92.047.9～95.650.7～98.352.2～100.153.8～102.63846.0～91.747.5～96.248.9～98.551.9～101.053.4～102.955.0～105.756.6～107.73948.5～98.550.0～101.153.0～103.654.6～106.256.3～108.857.9～111.459.6～113.84051.2～103.652.8～106.256.0～109.057.7～111.759.4～114.661.1～117.262.9～120.24154.1～109.055.8～111.857.5～114.960.9～118.062.8～121.064.6～124.166.5～126.34257.3～115.459.1～118.360.9～121.962.8～125.066.5～128.068.4～131.770.4～135.34360.7～123.162.7～126.064.6～129.168.5～132.170.5～136.772.7～140.474.8～143.94464.6～130.666.6～135.368.7～138.770.9～142.275.1～145.477.4～150.079.8～154.04568.8～140.471.1～144.273.4～148.975.7～152.678.1～156.680.6～160.783.1～165.24671.6～151.776.1～155.978.7～159.981.3～163.983.9～168.386.7～176.889.5～181.94779.3～163.782.0～168.084.8～177.085.3～181.988.2～187.291.3～192.797.2～198.84883.5～181.986.5～187.289.7～192.992.9～199.099.2～206.3102.8～212.4106.7～222.44988.7～203.492.1～209.398.7～219.3102.6～228.4106.8～235.5111.2～243.4115.9～260.5大孔陽性格數(shù)小孔陽性格數(shù)78910121302.9～13.73.7～15.34.5～15.85.2～16.95.9～18.56.9～20.17.8～21.213.7～15.34.3～16.25.2～17.26.0～18.56.8～20.17.6～21.78.7～22.224.4～16.95.3～17.86.0～18.66.8～20.17.7～21.78.5～22.99.4～24.035.1～18.36.1～18.86.9～20.27.7～21.88.6～23.69.5～24.110.2～25.345.9～19.56.9～20.57.8～21.88.7～23.49.6～24.610.3～25.710.9～26.956.8～21.27.6～21.98.5～23.49.4～25.110.4～26.411.0～27.011.7～28.667.7～22.08.6～23.69.5～25.110.5～26.811.1～27.511.8～28.812.5～30.478.7～24.09.6～25.310.6～26.911.6～28.212.3～29.312.6～30.513.7～31.989.7～25.7104～26.911.4～28.612.4～30.013.1～30.713.9～32.314.6～33.8910.5～27.011.5～28.612.6～30.413.3～31.214.0～32.614.7～34.215.9～35.21011.6～28.812.7～30.413.4～31.814.1～33.114.9～34.516.1～35.716.8～37.112.4～30.713.6～32.314.3～33.815.5～35.015.8～36.117.0～37.718.3～39.11213.7～32.614.9～34.215.7～35.215.9～36.517.2～38.118.0～39.219.3～40.61314.6～34.515.8～35.716.6～37.117.4～38.518.2～39.919.5～41.320.3～42.71416.0～36.116.8～37.717.6～39.118.4～40.619.7～41.820.6～43.422.0～44.41517.0～38.117.8～39.718.6～41.219.4～42.320.8～44.021.6～44.923.1～46.51618.0～40.218.8～41.619.6～42.721.0～44.421.9～45.723.4～47.224.3～48.81719.0～42.020.4～43.421.3～44.922.2～46.527.3～48.024.6～49.525.5～51.01820.7～44.221.6～45.722.4～47.223.3～48.824.9～50.325.8～51.226.7～52.81921.8～46.522.7～48.023.6～49.525.2～51.026.1～52.027.0～53.727.9～55.42023.0～48.823.9～50.325.5～51.226.4～52.827.4～54.528.3～56.230.1～57.62124.2～51.025.8～52.026.8～53.727.7～55.428.7～57.130.5～58.631.5～60.12226.2～52.927.1～54.528.1～56.229.1～58.030.9～59.731.9～60.832.9～62.52326.7～55.428.5～57.129.5～58.930.5～60.232.3～61.833.4～63.634.4～65.42428.9～58.029.9～59.830.9～61.332.8～63.033.9～64.734.9～66.136.0～67.8大孔陽性格數(shù)小孔陽性格數(shù)7891012132530.3～60.731.3～62.533.3～64.234.4～66.035.4～67.336.5～69.037.6～70.72631.8～63.532.8～65.434.9～66.636.0～68.537.1～70.438.2～72.240.4～73.72733.3～66.135.4～67.836.5～69.737.6～71.738.8～73.441.0～75.042.2～77.22835.0～69.137.1～71.038.2～73.039.4～74.441.7～76.441.7～77.944.1～80.02936.6～72.338.8～74.140.0～75.941.2～77.842.4～79.544.8～81.446.0～83.33039.5～75.840.7～77.641.9～79.543.1～81.445.6～83.346.8～85.348.1～87.33141.4～79.542.6～81.443.9～83.346.4～85.347.7～87.349.0～89.550.3～91.63243.4～83.344.7～85.346.0～87.348.6～89.549.9～91.651.3～93.952.6～94.63345.5～87.246.8～89.249.5～91.650.9～93.952.3～94.653.7～96.255.1～98.53447.7～91.550.5～93.850.5～94.651.9～96.254.8～98.556.2～101.057.7～103.63550.1～94.651.5～96.453.0～98.855.9～101.257.4～103.659.0～106.260.5～109.03652.6～99.554.1～101.455.6～103.858.7～106.460.3～109.161.9～111.863.5～114.73755.3～104.556.9～107.060.0～109.461.7～112.263.3～114.965.0～117.566.7～120.23858.2～110.359.8～112.863.2～115.464.9～118.366.6～121.068.4～123.170.2～126.03961.3～115.563.0～119.266.6～121.668.4～124.270.2～127.672.1～130.674.0～133.74066.5～123.166.5～125.470.2～128.972.2～132.174.1～135.076.1～137.078.2～140.44168.4～130.572.3～133.774.3～136.176.4～139.378.5～142.780.6～146.282.8～149.14272.5～138.274.6～142.178.8～145.481.0～148.883.3～151.785.6～155.987.9～159.84377.0～147.279.3～151.781.6～155.586.3～158.288.7～163.391.2～167.893.8～172.14482.2～158.184.7～162.787.2～167.292.2～172.095.0～174.495.1～177.997.9～182.94585.7～170.190.8～176.493.6～180.496.5～184.099.5～190.0102.6～195.0105.7～199.94692.4～186.998.1～192.3101.3～196.8104.6～201.1108.0～207.8111.6～215.4115.2～220.647100.5～204.8104.0～211.8107.7～218.7111.4～226.4115.4～234.0119.5～242.2123.8～250.548110.7～229.7114.9～238.0119.4～245.0124.0～257.8125.3～268.8130.4～279.5135.7～284.049120.8～270.3126.1～281.0128.0～292.9137.9～306.9140.2～320.9147.0～343.5159.0～360.0大孔陽性格數(shù)小孔陽性格數(shù)1415161718192008.6～21.99.0～23.49.6～24.910.5～25.711.5～26.912.5～28.613.2～29.319.4～23.69.7～25.110.6～26.411.6～27.012.6～28.613.3～30.014.0～30.729.8～25.110.4～26.811.4～27.512.4～28.813.5～30.414.1～31.214.8～32.6310.5～26.911.5～28.212.5～29.213.6～30.414.3～31.815.0～32.715.7～34.2411.6～28.612.6～29.313.7～30.714.8～32.215.1～33.115.8～34.517.0～35.7512.4～30.013.4～31.214.6～32.315.7～33.816.0～35.017.2～36.117.9～37.5613.6～31.814.7～32.615.9～34.216.6～35.217.4～36.518.1～37.819.4～39.1714.9～33.115.6～34.516.8～35.717.5～37.118.3～38.119.6～39.720.4～40.6815.7～35.017.0～36.117.7～37.718.5～38.519.2～40.220.6～41.321.4～42.7917.1～36.517.9～38.118.7～39.219.5～40.620.8～41.821.6～43.222.4～44.41018.1～38.518.9～39.719.7～41.221.0～42.321.8～43.423.3～44.923.5～46.519.1～40.219.9～41.620.7～42.722.1～44.223.6～45.723.7～46.825.2～48.01220.1～42.021.5～43.422.3～44.923.2～46.224.7～47.224.8～48.826.4～50.31321.8～44.222.6～45.623.4～46.625.0～48.025.8～49.526.7～51.027.6～52.01422.9～45.823.7～47.224.6～48.826.1～50.327.0～51.227.9～52.828.8～54.51524.0～48.024.9～49.526.4～51.027.3～52.028.2～53.729.1～55.230.9～56.21625.2～50.326.8～51.227.7～52.828.6～54.529.5～55.930.4～57.132.2～58.61726.4～52.028.0～53.728.9～55.429.9～56.430.8～58.032.6～59.733.6～60.81827.6～54.529.3～56.130.3～57.431.2～58.933.0～60.234.0～61.835.0～63.41929.7～57.030.7～58.031.6～59.832.6～61.334.5～62.635.5～64.236.5～66.02031.1～59.032.0～60.733.0～62.534.9～63.636.0～65.437.0～66.638.0～68.52132.5～61.633.5～63.035.4～64.736.5～66.137.5～67.838.5～69.239.6～70.92233.9～64.235.0～66.037.0～67.338.0～69.039.1～70.440.2～72.241.2～73.72335.5～66.637.5～68.538.6～70.339.7～71.740.8～73.441.9～75.043.0～76.42438.1～69.739.2～71.040.3～73.041.4～74.442.5～75.944.8～77.845.9～79.5大孔陽性格數(shù)小孔陽性格數(shù)141516171819202539.7～72.440.9～74.042.0～75.943.1～77.645.5～79.546.7～81.247.8～83.02641.5～75.742.7～77.643.8～79.245.0～80.846.2～82.748.6～84.249.8～85.42743.3～78.644.5～80.645.7～82.346.9～83.749.4～85.350.7～87.351.9～89.52845.3～81.546.5～83.447.7～85.350.3～87.351.6～89.552.8～91.654.1～93.92947.3～85.348.6～87.351.2～89.551.1～91.653.8～93.955.1～94.656.5～96.23049.4～89.550.7～91.652.0～93.954.8～94.656.1～96.257.5～98.658.9～101.03151.7～93.953.0～94.655.8～96.257.2～98.558.6～101.060.1～103.661.5～105.73254.0～96.256.9～98.558.4～101.059.8～103.661.3～106.262.8～108.064.3～109.43358.1～101.059.5～103.661.0～106.262.6～108.364.1～110.365.6～112.267.2～114.93460.8～106.262.3～108.863.9～111.165.5～112.867.1～115.168.7～118.070.3～120.43562.1～111.465.3～113.867.0～115.468.6～118.370.3～121.272.0～123.173.7～126.03666.9～116.468.6～119.270.3～121.872.0～124.273.8～127.175.5～129.177.3～132.13770.3～121.972.0～125.073.8～127.675.7～130.677.5～133.779.4～135.383.4～138.73873.9～128.975.8～132.177.7～135.079.7～137.081.6～140.483.6～143.085.6～146.23975.9～135.377.9～138.782.0～142.284.0～145.486.1～148.288.3～151.792.8～154.74080.2～144.084.5～147.986.7～150.088.9～154.091.1～156.695.9～159.995.8～163.74187.2～152.689.6～156.191.9～159.894.3～163.794.2～168.099.2～172.4101.8～174.44292.8～163.795.3～168.095.3～172.497.8～174.4103.1～177.0105.9～182.0108.7～187.24396.5～17