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铜离子加银离子与游离氯协同 灭活水中病毒效果的试验观察
http://www.100md.com 《中国消毒学杂志》 1998年第1期
     作者:骆文静 张 进 胡艳冰 张文清 侯 悦

    单位:第四军医大学军队卫生教研室, 西安 710033

    关键词:协同灭活作用;游离氯;铜离子;银离子;大肠杆菌噬菌体f;2;脊髓灰质炎病毒I型

    中国消毒学杂志980105 提要 铜离子或银离子对水中大肠杆菌噬菌体f 2 与脊髓灰质炎病毒I型灭活效果极低。但以400 μg/L铜离子加40 μg/L银离子与0.3 mg/L游离氯协同作用30 min,则可将两者存活率对数分别减少至4.806与2.674,远较单独使用游离氯效果为优。

    游离氯灭活水中病毒所需浓度较高,有较浓氯味。为减少游离氯用量,对铜、银离子与其协同灭活水中病毒的效果进行了试验观察。现将结果报道如下。

, http://www.100md.com     1 方法

    1.1 病毒悬液的制备

    以大肠杆菌(285)为宿主菌,制备大肠杆菌噬菌体f 2 原液。经40 000 r/min离心4 h,取沉淀,用灭菌的双蒸去离子水配成噬菌体f 2 悬液(2.0×10 5 ~2.5×10 5 pfu/ml。

    将脊髓灰质炎病毒Ⅰ型(PV-Ⅰ)接种于人表皮样癌细胞(He P-2细胞)。培养5~7 d,收获病毒,用聚乙二醇-6000沉淀并结合差速离心,浓缩纯化。然后,加入灭菌的双蒸去离子水,制成含量为3.0×10 3 ~5.5×10 4 pfu/ml的病毒悬液。

    1.2 病毒灭活试验

    向含噬菌体f 2 或PV-Ⅰ的磷酸盐缓冲液(pH 7.2~7.4,温度为20~25℃)中加入预定浓度的消毒剂,混匀。作用30 min,取10 ml混合液,加入含0.5 ml中和剂(14.6%硫代硫酸钠、10.0%硫代乙醇酸钠溶液)的试管中。混匀并中和后,接种于宿主菌或细胞,培养计数。按下式计算存活率对数:
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    K = -lg (N t /N 0 )

    〔 N t 为消毒作用后存活病毒数;N 0 为消毒前活病毒数。〕

    2 结果

    2.1 游离氯对病毒的灭活作用

    试验表明,在水中的游离氯对噬菌体f 2 和PV-Ⅰ的灭活作用,随其浓度升高而增强。以0.3 mg/L游离氯作用30 min,使噬菌体f 2 和PV-Ⅰ存活率对数分别减少2.839与1.125;以1.0 mg/L游离氯作用30 min,噬菌体f 2 存活率对数可减少4.500,PV-Ⅰ者为2.502(表1)。

    2.2 铜与银离子对病毒的灭活作用

    以500 μg/L铜作用30 min,仅使噬菌体f 2 与PV-Ⅰ存活率对数分别减少0.110与0.059;以50 μg/L银作用者亦仅分别为0.192与0.086。500 μg/L铜与50 μg/L银同时作用30 min,可使噬菌体f 2 与PV-Ⅰ存活率对数分别减少0.243与0.155(表2)。
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    2.3 铜加银离子与游离氯协同对病毒的灭活作用

    以铜加银离子与游离氯协同,对噬菌体f 2 与PV-Ⅰ的灭活作用强于以铜加银离子或单独游离氯作用者。以400 μg/L铜、40 μg/L银与0.3 mg/L游离氯协同作用30 min,可将噬菌体f 2 与PV-Ⅰ存活率对数分别减少4.806与2.674(表3)。

    表 1 不同浓度游离氯对水中病毒的灭活效果

    Table 1 Efficacy of free chlorine at different con- centrations in inactivating viruses in water 游离氯浓度

    Concentration of Longarithm of survival rate free chlorine(mg/L)
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    存活率对数(K)

    Longarithm of survival rate

    嗜菌体f 2

    Bacteriophage f 2

    PV-Ⅰ

    0.0

    -0.782

    -0.192

    0.2

    -2.109

    -0.362

    0.3
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    -2.839

    -1.125

    0.4

    -3.257

    -1.340

    1.0

    -4.500

    -2.502

    注: 阳性对照组嗜菌体f 2 含量为250 000 pfu/ml,PV-Ⅰ 含量为4 000 pfu/ml。 试验温度为20~25℃。作用 时间为30 min。

    Note: Bacteriophage f 2 content was 250 000 pfu/ml and PV-Ⅰ content was 4 000 pfu/ml in positive control group. The temperature was 20~25℃. Contact time was 30 min.
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    3 结语

    结果表明,用50 μg/L银离子对水中大肠杆菌噬菌体f 2 与PV-Ⅰ作用30 min,仅使其存活率对数分别减少0.192与0.086;以500 μg/L铜离子作用30 min,亦仅使其存活率对数分别减少0.110与0.059;经0.3 mg/L游离氯作用30 min,可使其存活率对数分别减少2.839与1.125。然而,以400 μg/L铜离子加40 μg/L银离子与0.3 mg/L游离氯协同作用30 min,则可将水中噬菌体f 2 与PV-Ⅰ存活率对数分别减少4.806与2.674。关于铜与银离子协同提高游离氯灭活病毒的作用机理,尚待进一步研究。

    表 2 铜与银离子对水中病毒的灭活效果

    Table 2 Efficacy of copper ion and silver ion in inactivating viruses in water 金属离子浓度
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    存活率对数(K)

    Longariophagef2 survival rate

    Concentration of Longarithm of survival rate metal ion(μg/L)

    嗜菌体f 2

    Bacteriophage f 2 铜 Copper:

    PV-Ⅰ

    铜Copper:

    100

    -0.017
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    -0.002

    200

    -0.040

    -0.011

    300

    -0.061

    -0.019

    400

    -0.083

    -0.042

    500

    -0.110

    -0.059
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    银Silver:

    10

    -0.044

    -0.004

    20

    -0.070

    -0.019

    30

    -0.102

    -0.054

    40

    -0.114

    -0.066
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    50

    -0.192

    -0.086

    铜/银Copper/Silver:

    200/20

    -0.040...

    300/30

    -0.111

    -0.030

    400/40

    -0.222

    -0.103
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    500/50

    -0.243

    -0.155

    注: 阳性对照组嗜菌体f 2 含量为200 000 pfu/ml,PV-I 含量为3 000 pfu/ml。试验温度为20~25℃。作用 时间为30 min。

    Note: Bacteriophage f 2 content was 200 000 pfu/ml and PV-Ⅰ content was 3 000 pfu/ml in positive control group. The temperature was 20~25℃. Contact time was 30 min.

    表 3 铜及银离子与游离氯协同对水中病毒的灭活效果
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    Table 3 Synergetic efficacy of copper ion, silver ion and free chlorine in inactivating viruses in water

    消毒剂浓度 Disinfectant and concentration

    存活率对数 Longarithm of survival rate (K)

    嗜菌体f 2

    Bacteriophage f 2

    PV-Ⅰ

    游离氯 (0.3 mg/L)

    Free chlorine铜 (400 μg/L) + 银 (40 μg/L)
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    -2.839

    -1.125

    Copper ion (400 μg/L) + silver ion (40 μg/L)

    -0.222

    -0.103

    铜 (400 μg/L) + 银 (40 μg/L) + 游离氯 (0.3 mg/L)

    Copper ion (400 μg/L)+silver ion (40 μg/L)+free chlorine (0.3 mg/L)

    -4.806

    -2.674
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    注: 阳性对照组嗜菌体f 2 含量为250 000 pfu/ml,PV-Ⅰ 含量为55 000 pfu/ml。试验温度为20~25℃。作用时间为30 min。

    Note: Bacteriophage f 2 content was 250 000 pfu/ml and PV-Ⅰ content was 55 000 pfu/ml in positive control group. The temperature was 20~25℃. Contact time was 30 min. EXPERIMENTAL OBSERVATION ON SYNERGETIC EFFICACY OF COPPER ION, SILVER ION AND FREE CHLORINE IN INACTIVATING VIRUSES IN WATER

    Luo Wenjing Zhang Jin Hu Yanbing Zhang Wenqing Hou Yue ( Department of Military Hygiene, Fourth Military Medical University, Xi'an 710033 )
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    Abstract

    In order to reduce free chlorine concentration required for inactivating viruses in water and thus to reduce chlorine odor, the synergetic efficacy of copper ion, silver ion and free chlorine in inactivating viruses in water was observed experimentally.

    The results indicated that 50 μg/L silver ion with a 30 min contact time could reduce the logarithms of survival rates of bacteriophage f 2 of Escherichia coli and poliomyelitis virus type Ⅰ (PV-Ⅰ) in water only by 0.192 and 0.086 respectively; 500 μg/L copper ion with a 30 min contact time also could only reduce them by 0.110 and 0.059 respectively; 0.3 mg/L free chlorine with a 30 min contact time could reduce them by 2.839 and 1.125 respectively. However, 400 μg/L copper ion plus 40 μg/L silver ion and 0.3 mg/L free chlorine with a 30 min contact time could reduce the logarithms of survival rates of bacteriophage f 2 and PV-Ⅰ in water by 4.806 and 2.674 respectively.

    Key words synergetic inactivating action free chlorine copper ion silver ion bacteriophage f 2 of Escherichia coli poliomyelitis virus type Ⅰ

    (本工作承高双斌高级实验师相助,特此致谢。)

    (1996-10-15收稿 1997-09-22修回), 百拇医药