4种常用消毒剂对H7N9亚型流感病毒灭活效果研究

为了比较常用消毒剂对H7N9亚型流感病毒杀灭效果,依照2002年版《消毒技术规范》,评估过硫酸氢钾复合物粉、聚维酮碘溶液、戊二醛癸甲溴铵溶液、稀戊二醛溶液4种消毒剂在不同浓度下,与H7N9亚型流感病毒作用10 min、30 min和60 min的灭活效果。结果显示,0.2%(w/v)过硫酸氢钾复合物粉溶液作用10 min,0.5%(w/v)聚维酮碘溶液作用30 min,0.1%(v/v)戊二醛癸甲溴铵溶液作用30 min,或0.3%(v/v)稀戊二醛溶液作用30 min,均可将H7N9亚型流感病毒完全灭活。与推荐使用浓度相比,戊二醛癸甲溴铵溶液、稀戊二醛溶液、聚维酮碘可以有效灭活H7N9亚型流感病毒,而过硫酸氢钾复合物粉不能有效灭活H7N9亚型流感病毒。本研究对H7N9亚型流感病毒消毒剂的选用具有指导意义。     

3种常用消毒剂对基因Ⅶ型新城疫病毒的灭活效果

为评价种禽场常用消毒剂对基因Ⅶ型新城疫病毒(NDV)的灭活效果,采用悬液定量法和鸡胚接种试验对癸甲溴铵溶液、聚维酮碘溶液和戊二醛苯扎溴铵溶液3种消毒剂的最低杀毒浓度及其工作液的稳定性进行了研究。在试验条件下,3种消毒剂对基因Ⅶ型NDV均有良好的杀灭效果。癸甲溴铵、聚维酮碘和戊二醛苯扎溴铵溶液杀灭病毒的最低有效浓度分别为25、44.44 mg/L和0.27 mL/L(含戊二醛和苯扎溴铵各约26.67 mg/L)。配制的5 g/L聚维酮碘溶液和0.1 g/L癸甲溴铵溶液消毒工作液完全杀灭病毒的时间分别可保持10 d和135 d,而10 mL/L戊二醛苯扎溴铵溶液则至少可保持180 d。     

几种消毒剂消毒效果观察

本文针对鸡场常见的4种主要致病茵（大肠杆菌、沙门氏菌、巴氏杆菌、葡萄球菌）和1种病毒（NDV病毒）,选取13种消毒剂,进行细菌和病毒杀灭比较实验。结果表明：三氯异氰尿酸粉,碘＋磷酸溶液（2）,癸甲溴铵溶液（2）,过氧化氢＋过氧乙酸在低浓度下作用10min即可杀死新城疫病毒和4种主要致病细菌,为综合效果最好的消毒药;癸甲溴铵溶液（1）,碘＋磷酸溶液（1）,戊二醛甲溴铵溶液．复合酚消毒剂（1）,复合酚消毒剂（2）,复合酚消毒剂（3）应适当提高浓度或延长时间才能有效消毒;月苄三甲氯铵溶液,癸甲溴铵溶液（3）,癸甲溴铵＋碘溶液在任一推荐浓度下30min内均存在杀细菌或杀病毒无效的情况。     

养殖场消毒池常用消毒剂杀菌效能对比

通过模拟消毒池状况,对当前消毒池四种常用消毒剂(烧碱、二氯异氰脲酸钠粉、戊二醛癸甲溴铵溶液、聚维酮碘溶液)进行杀菌对比试验。文章通过试验对比和分析,推荐在养殖场消毒池使用30%消毒威(通用名:二氯异氰脲酸钠粉)1:1200倍兑水稀释液、新菌敌(通用名:戊二醛癸甲溴铵溶液)1:500倍兑水稀释液进行车轮和鞋底消毒,每5~6天更换1次;不建议使用烧碱(Na OH)和聚维酮碘溶液消毒。     

戊二醛癸甲溴铵溶液对HP-PRRSV体外杀灭效果测定

针对高致病性蓝耳病病毒（PRRSV）,农业部动物疫病防控重点开放实验室用戊二醛癸甲溴铵溶液对其进行了体外杀灭效果试验,结果显示大于或等于0.01%浓度的戊二醛癸甲溴铵溶液可体外完全杀灭PRRSV。根据结果建议该消毒剂对PRRSV临床使用浓度为≥0.01%,即1∶10 000稀释,但同时也要根据具体情况具体分析,适当提高使用浓度。     

猪种布鲁氏菌对4种常用新型消毒剂敏感性的研究

为了确定常用新型消毒剂对布鲁氏菌的杀菌/抑菌效果,本研究将聚维酮碘、戊二醛、戊二醛癸甲溴铵复方溶液、二氯异氰尿酸钠4种常用消毒剂与定量B.suis混合均匀,分别作用3 min、10 min与30 min,取0.1 m L涂布于TSA培养基中进行菌落计数;通过对不同消毒剂、不同作用时间其杀菌率的对比,确定不同消毒剂对B.suis的敏感程度。研究表明,B.suis对聚维酮碘溶液敏感性最强,10%聚维酮碘溶液1 000倍稀释作用3 min便能有效杀灭B.suis;其次戊二醛与癸甲溴铵混合溶液效果显著优于戊二醛单独使用,100倍稀释后可有效杀灭B.suis,1 000倍稀释后作用30 min可100%杀灭B.suis;而单独使用戊二醛,仅在10倍稀释时3~30 min能有效杀灭该菌,100倍稀释时作用30 min,其杀菌率仅达93%;而二氯异氰尿酸钠在4种消毒剂中消毒效果最不理想,其最高杀菌率仅为67%;因此,在布鲁氏菌病的防控中,消毒剂以聚维酮碘与戊二醛癸甲溴铵复方制剂为最佳选择。     

戊二醛苯扎溴铵溶液对H9N2亚型禽流感病毒的灭活效果分析

采用悬浮杀灭试验,考察戊二醛苯扎溴铵溶液对H9N2亚型禽流感病毒的灭活效果。将戊二醛苯扎溴铵溶液通过不同稀释倍数(1 250、2 500、5 000、10 000、20 000)、在不同作用时间(5、10 min)观察其对H9N2亚型禽流感病毒的灭活效果。结果显示,戊二醛苯扎溴铵溶液在10000倍稀释,与H9N2亚型禽流感病毒作用5 min或10 min,灭活率达99.90%以上;在20 000倍稀释,作用5 min或10 min,灭活率均小于99.90%。表明戊二醛苯扎溴铵溶液在10 000倍稀释与H9N2亚型禽流感病毒(20±1)℃水浴作用5 min,即可对H9N2亚型禽流感病毒达到很好的灭活效果。     

不同消毒剂对PRV、PPV和TGEV的灭活效果试验

为了确定癸甲溴铵-碘溶液消毒剂和癸甲溴铵消毒剂对猪主要疫病病原如猪细小病毒(PPV)、伪狂犬病病毒(PRV)和猪传染性胃肠炎病毒(TGEV)的灭活效果,通过细胞观察法,用不同浓度的癸甲溴铵-碘溶液消毒剂和癸甲溴铵消毒剂对PRV、PPV和TGEV进行灭活来观察其灭活效果。结果表明,癸甲溴铵-碘溶液和癸甲溴铵溶液对细胞的最大无毒剂量分别为1∶80和1∶10。1∶320的癸甲溴铵-碘溶液消毒剂,1∶80的癸甲溴铵消毒剂与相应浓度的中和剂的反应产物已无毒副作用。癸甲溴铵-碘溶液消毒剂1∶800稀释时可使PRV TCID₅₀降低5个滴度,TGEV TCID₅₀降低4个滴度;癸甲溴铵消毒剂稀释到1∶400时,可使PRV和TGEV的TCID₅₀均降低约3个滴度。两种消毒剂对PPV的灭活效果均不理想。     

不同浓度格利特消毒剂对鸡粪中H9N2亚型禽流感病毒杀灭效果观察

为验证在有机物存在下格利特(20％戊二醛溶液)消毒剂杀灭禽流感病毒的能力,试验模拟空栏鸡舍和周围环境消毒实际情况,在鸡粪中混入H9亚型禽流感病毒,以接种鸡胚尿囊液中H9亚型禽流感病毒血凝价＜2 log2判为感染阴性,以样品经消毒剂作用后检测不到病毒含量(即0EID50)为病毒完全杀灭(100％杀灭病毒),研究格利特消毒剂杀灭禽流感病毒的效果,结果显示:1∶400以下稀释的格利特消毒剂4℃或25℃作用20 min,均能100％杀灭混入鸡粪中的H9N2禽流感病毒;1∶200稀释的格利特消毒剂25℃作用20 min,能100％杀灭存在于鸡粪和清洁剂中的H9N2禽流感病毒.1∶1600以下稀释的格利特消毒剂25℃作用20 min,均能100％杀灭无鸡粪保护的H9N2禽流感病毒.     

三种常用消毒剂对大肠杆菌的杀菌效果试验

以大肠杆菌为杀菌对象,通过中和试验、悬液定量杀菌试验和定性杀菌试验评价戊二醛癸甲溴铵溶液、聚维酮碘、月苄三甲氯铵三种消毒剂的杀菌效果。结果表明:当三种消毒剂溶液浓度达到MBC(最小杀菌浓度)时,作用3 min对大肠杆菌的杀菌率为100%;三种消毒剂分别按1:256、1:512倍稀释并作用5 min,戊二醛癸甲溴铵溶液的杀菌效果优于其它两种消毒剂。     

An H5N1 highly pathogenic avian influenza virus that invaded Japan through waterfowl migration

In 2010, an H5N1 highly pathogenic avian influenza virus (HPAIV) was isolated from feces of apparently healthy ducks migrating southward in Hokkaido, the northernmost prefecture of Japan. The H5N1 HPAIVs were subsequently detected in domestic and wild birds at multiple sites corresponding to the flyway of the waterfowl having stopovers in the Japanese archipelago. The Hokkaido isolate was genetically nearly identical to H5N1 HPAIVs isolated from swans in the spring of 2009 and 2010 in Mongolia, but less pathogenic in experimentally infected ducks than the 2009 Mongolian isolate. These findings suggest that H5N1 HPAIVs with relatively mild pathogenicity might be selected and harbored in the waterfowl population during the 2009-2010 migration seasons. Our data provide "early warning" signals for preparedness against the unprecedented situation in which the waterfowl reservoirs serve as perpetual sources and disseminators of HPAIVs.     

三类常用消毒剂对非洲猪瘟病毒灭活效果的评价

为评价戊二醛类、酚类、含氯类常用消毒剂对非洲猪瘟病毒(ASFV)的灭活效果,参考OIE参考实验室相关操作流程,基于畜禽栏舍、运载工具、器具消毒目的,根据说明书标明的浓度范围选择低、中、高3个工作浓度,与ASFV分别在4℃和20℃条件下作用30 min,10倍连续稀释后接种猪肺泡巨噬细胞,同时加入猪红细胞,培养观察红细胞...     

Genetic relationship of H3 subtype avian influenza viruses isolated from domestic ducks and wild birds in Korea and their pathogenic potential in chickens and ducks

The H3 subtype avian influenza virus (AIV) is one of the most frequently isolated subtypes in domestic ducks, live poultry markets, and wild birds in Korea. In 2002-2009, a total of 45 H3 subtype AIVs were isolated from the feces of clinically normal domestic ducks (n=28) and wild birds (n=17). The most prevalent subtypes in domestic ducks were H3N2 (35.7%), H3N6 (35.7%), H3N8 (25.0%), and H3N1 (3.6%, novel subtype in domestic duck in Korea). In contrast, H3N8 (70.6%) is the most prevalent subtype in wild birds in Korea. In the phylogenetic analysis, HA genes of the Korean H3 AIVs were divided into 3 groups (Korean duck, wild bird 1, and wild bird 2) and all viruses of duck origin except one were clustered in a single group. However, other genes showed extensive diversity and at least 17 genotypes were circulating in domestic ducks in Korea. When the analysis expanded to viruses of wild bird origin, the genetic diversity of Korean H3 AIVs became more complicated. Extensive reassortments may have occurred in H3 subtype influenza viruses in Korea. When we inoculated chickens and ducks with six selected viruses, some of the viruses replicated efficiently without pre-adaptation and shed a significant amount of viruses through oropharyngeal and cloacal routes. This raised concerns that H3 subtype AIV could be a new subtype in chickens in Korea. Continuous surveillance is needed to prepare the advent of a novel subtype AIV in Korea.     

Molecular epidemiology of avian influenza viruses circulating among healthy poultry flocks in farms in northern Vietnam

Repeated epizootics of highly pathogenic avian influenza (HPAI) virus subtype H5N1 were reported from 2003 to 2005 among poultry in Vietnam. More than 200 million birds were killed to control the spread of the disease. Human cases of H5N1 infection have been sporadically reported in an area where repeated H5N1 outbreaks among birds had occurred. Subtype H5N1 strains are established as endemic among poultry in Vietnam, however, insights into how avian influenza viruses including the H5N1 subtype are maintained in endemic areas is not clear. In order to determine the prevalence of different avian influenza viruses (AIVs), including H5N1 circulating among poultry in northern Vietnam, surveillance was conducted during the years 2006-2009. A subtype H5N1 strain was isolated from an apparently healthy duck reared on a farm in northern Vietnam in 2008 and was identified as an HPAI. Although only one H5N1 virus was isolated, it supports the view that healthy domestic ducks play a pivotal role in maintaining and transmitting H5N1 viruses which cause disease outbreaks in northern Vietnam. In addition, a total of 26 AIVs with low pathogenicity were isolated from poultry and phylogenetic analysis of all the eight gene segments revealed their diverse genetical backgrounds, implying that reassortments have occurred frequently among strains in northern Vietnam. It is, therefore, important to monitor the prevalence of influenza viruses among healthy poultry between epidemics in an area where AIVs are endemic.     

两株H4N3禽流感病毒全基因组序列分析及对小鼠的致病性研究

2009年在我国南方活禽交易市场进行流行病学调查时,从鸭体内分离到2株H4N3亚型禽流感病毒(AIV),DK/FJ/S1419/09(H4N3)(FJ/419/2009)和DK/HUN/S1010/09(H4N3)(HuN/010/2009)。为了解这2株H4N3亚型AIV的生物学特性,本研究对其进行全基因组分析及对小鼠致病性研究。结果显示:其HA基因来源于近两年流行的H4亚型病毒株,NA基因来源于其它亚型病毒株。内部基因来源较复杂,与FJ/419/2009内部基因同源性最高的病毒株均来自国内H5、H6等亚型分离株,但与HuN/010/2009同源的内部基因则差异较大,与PA、M和NS同源性最高的病毒株分别为A/wild/duck/Korea/UP122/2007(H1N1),A/muscovy/duck/Thailand/CU-LM1983/2009(H4N6)and A/avian/Japan 8KI0068/2008(H3N6)。用106EID50病毒剂量感染6周龄BALB/c小鼠,结果显示试验组小鼠感染后第3 d采脏器样品,仅在鼻甲和肺部能检测到病毒存在。以上数据表明,尽管这2株H4N3特殊亚型组合病毒来源复杂,但对小鼠的致病性较低。     

The first specific detection of a highly pathogenic avian influenza virus (H5N1) in Ivory Coast

The Virology Laboratory of the Central Laboratory of Animal Diseases in Ivory Coast at Bingerville received samples of wild and domestic avian species between February and December 2006. An RT-PCR technique was used to test for avian influenza (AI) and highly pathogenic AI subtype viruses. Among 2125 samples, 16 were type A positive; of which, 12 were later confirmed to be H5N1. Fifteen of these 16 type A positive samples were inoculated into the chorioallantoic cavity of 11-day-old embryonated hens' eggs for virus isolation. Eight produced virus with hemagglutination titres from 1/64 to 1/512. The 4/16 M-RT-PCR positive samples, which were H5N1 negative, were shown to be H7 subtype negative. The diagnostic efficiency of the laboratory for the surveillance of H5N1 in Ivory Coast was demonstrated. The positive cases of H5N1 were from a sparrowhawk (Accipter nisus); live market poultry and in free-range poultry, where the mortality rate was approximately 20% (2/10) and 96.7% (29/30) respectively. Currently, investigations into intensive poultry farms have proved negative for H5N1. No human cases have been reported this time.     

Putative human and avian risk factors for avian influenza virus infections in backyard poultry in Egypt

Highly pathogenic influenza A virus subtype H5N1 causes significant poultry mortality in the six countries where it is endemic and can also infect humans. Egypt has reported the third highest number of poultry outbreaks (n = 1084) globally. The objective of this cross-sectional study was to identify putative risk factors for H5N1 infections in backyard poultry in 16 villages in Damietta, El Gharbia, Fayoum, and Menofia governorates from 2010–2012. Cloacal and tracheal swabs and serum samples from domestic (n = 1242) and wild birds (n = 807) were tested for H5N1 via RT-PCR and hemagglutination inhibition, respectively. We measured poultry rearing practices with questionnaires (n = 306 households) and contact rates among domestic and wild bird species with scan sampling. Domestic birds (chickens, ducks, and geese, n = 51) in three governorates tested positive for H5N1 by PCR or serology. A regression model identified a significant correlation between H5N1 in poultry and the practice of disposing of dead poultry and poultry feces in the garbage (F = 15.7, p < 0.0001). In addition, contact between domestic and wild birds was more frequent in villages where we detected H5N1 in backyard flocks (F = 29.5, p < 0.0001).     

Virological surveillance and phylogenetic analysis of the PB2 genes of influenza viruses isolated from wild water birds flying from their nesting lakes in Siberia to Hokkaido, Japan in autumn

Recent introduction of H5N1 highly pathogenic avian influenza virus (HPAIV) in wild birds from poultry in Eurasia signaled the possibility that this virus may perpetuate in nature. Surveillance of avian influenza especially in migratory birds, therefore, has been conducted to provide information on the viruses brought by them to Hokkaido, Japan, from their nesting lakes in Siberia in autumn. During 2008-2009, 62 influenza viruses of 21 different combinations of hemagglutinin (HA) and neuraminidase (NA) subtypes were isolated. Up to September 2010, no HPAIV has been found, indicating that H5N1 HPAIV has not perpetuated at least dominantly in the lakes where ducks nest in summer in Siberia. The PB2 genes of 54 influenza viruses out of 283 influenza viruses isolated in Hokkaido in 2000-2009 were phylogenetically analysed. None of the genes showed close relation to those of H5N1 HPAIVs that were detected in wild birds found dead in Eurasia on the way back to their northern territory in spring.