实验动物屏障系统微生物动态状况

目的研究运行中的实验动物屏障系统微生物的情况。方法采用沉降菌法、棉拭子法等方法,研究运行中的屏障系统不同区域、不同环境指标下屏障系统内微生物的状况。结果动态下的屏障系统微生物情况与国标GB14925-2001中静态环境有较大不同,动物饲养室和动物实验室沉降菌浓度远高于静态要求;辅助区域在规范化消毒及严格管理的情况下,能达到国标要求。屏障系统的微生物情况存在一定的昼夜变化规律,在晚间出现峰值。结论合适的换气次数可有效控制实验动物屏障系统的沉降菌浓度;加强消毒及硬件的管理,是屏障系统内环境稳定的保障。     

大型实验动物屏障设施环境微生物和尘埃粒子的动态监控

目的本实验以大型实验动物屏障设施为研究对象,分析屏障设施的环境微生物及尘埃粒子的动态变化规律及其相关影响因素。方法测定屏障设施各功能区域不同时间及不同工作状态下空气落下菌和直径≥0.3μm尘埃粒子的数量变化。结果屏障系统落下菌与尘埃粒子变化规律如下：屏障系统内空气落下菌与尘埃粒子在饲养工作后显著升高,喷雾消毒后明显降低;大、小鼠饲育室空气落下菌与尘埃数在凌晨时明显升高,而兔饲育室在凌晨时间段则较低;清洁走廊和污染走廊在工作状态时细菌含量明显上升,非工作状态时细菌含量一直处于较低的水平。结论实验动物屏障系统的环境微生物与尘埃粒子的动态数量变化与动物品种、空气消毒、人员进出及动物室内的饲养操作等有关。     

湖北省实验动物设施环境控制中存在的问题

目的分析总结湖北省2012年度实验动物许可单位年检中环境设施的检测结果,找出问题,提出促进湖北省实验动物环境设施可靠运行的建议。方法各实验动物环境设施采用30%区域抽检方式,依据GB14925-2010《实验动物环境及设施》,进行环境指标检测。结果除气流速度、沉降菌最大平均浓度外,其他技术指标均存在不符合国家标准的情况。结论应针对实验动物设施管理加强培训,提高实验动物设施管理人员的管理水平,从而保证实验动物设施的正常运行。     

医学实验动物屏障环境内独立通气笼具的安放

目的探讨屏障系统内安放独立通气笼盒（Individually Ventilated Cages,IVC）双保险模式饲养实验啮齿类动物,力求动物饲养、实验的全过程达到SPF级;降低动物实验室硬件建造费和维持费（节能）。方法通过改建一间屏障环境动物实验室,对空气洁净度等相关项目数据按GB14925-2001方法检定,并在该实验室内安放SPC级IVC饲养动物;在超净工作台内做打针投药等动物实验。结果改建的屏障环境动物实验室和IVC,各项检测数据均达到新国标要求。结论医学实验动物屏障环境内安放IVC饲养啮齿类动物、做动物实验,这种双保险模式能全过程达到SPF级要求;饲养、实验人员操作较简便,节能。     

洁净室环境监测用培养基贮存效期的验证及方法学确认

目的验证洁净室环境监测用培养基的贮存效期,同时对环境监测浮游菌、沉降菌及表面微生物检测方法进行确认。方法对连续3批次贮存0、90、120 d洁净室环境监测用胰酪大豆胨琼脂(Tryptose soya agar,TSA)培养皿和TSA(L-80)接触皿进行相应菌液适用性检查(包括促生长能力和无菌检查)验证试验;并对贮存120 d TSA培养皿和TSA(L-80)接触皿分别进行环境浮游菌和沉降菌、表面微生物最长采样时间检测,然后再进行促生长能力测试。结果 TSA培养皿和TSA(L-80)接触皿贮存120 d (2～8℃90 d,25℃30 d)的适用性检查结果均符合《中国药典》2015版(三部)对培养基质量控制的要求。贮存120 d TSA培养皿进行洁净室环境浮游菌(主动采样10 min)、沉降菌(暴露采样4 h),以及TSA(L-80)接触皿进行表面微生物(接触采样10 s)检测后均具有良好的促生长能力。结论通过验证试验,确定了环境监测用TSA培养皿和TSA(L-80)接触皿的贮存效期120 d(2～8℃90 d,25℃30 d),并确认了贮存后TSA培养皿进行洁净室环境浮游菌、沉降菌检测的有效性,以及TSA(L-80)接触皿进行表面微生物检测的有效性。     

象山港网箱养殖区与非养殖区的细菌群落分布

近海集约化养殖是导致我国近海污染的主要来源之一。浮游细菌在近海生态系统的物质循环过程中发挥重要作用,研究海洋浮游细菌群落对养殖活动的响应,对于指示和评价海水养殖生态系统健康具有重要意义。采集了象山港网箱养殖区与非养殖区3个深度,包括表层(0.5 m)、中层(2.5 m)、底层(8.0 m)的水样,利用焦磷酸测序技术测定16S rRNA基因,研究浮游细菌的群落结构和多样性。结果表明:网箱养殖活动不仅使得附近区域水体理化性质发生改变,如化学需氧量浓度显著高于非养殖区域,而且显著地(P0.05)改变了浮游细菌的群落结构,但不同深度间群落结构和多样性的差异不显著。网箱养殖区和非养殖区中主要浮游细菌类群为α-变形菌(Alphaproteobacteria)、γ-变形菌(Gammaproteobacteria)、拟杆菌(Bacteroidetes)、放线菌(Actinobacteria)、β-变形菌(Betaproteobacteria)、ε-变形菌(Epsilonproteobacteria)和其它变形菌(Unclassified Proteobacteria),占细菌总序列数的98.64%。有些细菌类群的平均相对丰度从网箱区到非养殖区差异显著,如拟杆菌(P0.01)和放线菌(P0.05)显著降低,而γ-变形菌(P0.05)显著增加。相似度分析表明γ-变形菌、α-变形菌和拟杆菌是造成网箱区和非养殖区群落差异的主要类群,对群落差异总贡献率达到45.02%。偏冗余分析表明,影响细菌群落分布的主要环境因子有化学需氧量、磷酸盐、铵盐和总有机碳,共解释38.18%的群落变异,空间距离单独解释10.66%的群落变异。实验结果表明,养殖活动导致浮游细菌群落的改变,其中拟杆菌、放线菌和γ-变形菌的丰度显著变化,可能用于评价养殖水体的水质状况。     

开放系统实验动物饲育室的合理换气问题

实验动物的繁殖饲养大都在相对窄小的笼具里,其中不仅有动物,还有排泄物。因此,实验动物对新鲜空气的要求很高。合理地控制实验动物饲育室的通风换气,将直接影响室内的清洁卫生及动物的健康,也关系到工作人员的健康。1 实验动物室的换气原则     

地鼠繁殖方式的改进与规模化生产

目的：探索SPF地鼠在屏障系统内的配种繁殖方式是否适合目前地鼠大规模生产,满足乙脑减毒活疫苗生产所需动物。方法：采取短期同居配种的繁殖方式进行试验和大规模生产。结果：短期同居配种的繁殖方式与人工刺激配种的繁殖方式相比具有仔鼠成活率高、受孕率高、单产高、利用率高的特点。结论：地鼠短期同居配种的繁殖方式适合在屏障系统内操作和大规模生产。     

肺炎克雷伯菌生物膜对小鼠巨噬细胞免疫功能的影响

目的研究肺炎克雷伯菌生物膜(BF)对小鼠腹腔巨噬细胞TLRs mRNA和细胞因子表达的影响,探索机体抗BF感染免疫的特点。方法将雄性昆明种小鼠40只随机分成2组,一组腹腔植入体外形成肺炎克雷伯菌BF的硅胶片,建立留置性医疗装置BF感染模型实验组,另一组植入与实验组同等量的浮游菌作为对照组。实时定量PCR分析2组巨噬细胞TLRs mRNA的表达水平,双抗体夹心ELISA法测定细胞因子的含量。结果实验BF组巨噬细胞TLR2、TLR4 mRNA表达量是对照浮游菌组的0.23和0.24倍;而TLR5、TLR9两组表达差异无显著性。实验BF组刺激前后IL-1、IL-2的差值明显低于对照浮游菌组,而IL-4则相反(P0.01)。结论与浮游菌相比,BF能下调小鼠腹腔巨噬细胞TLR2、TLR4的表达,机体的免疫应答朝着Th2型免疫反应发展,这可能是BF相对浮游菌更容易逃脱机体免疫防御系统、引起慢性感染的机制之一。     

铜绿假单胞菌生物膜和浮游菌的毒力表达差异

目的探究铜绿假单胞菌生物膜和浮游菌状态下毒力因子的表达差异。方法使用铜绿假单胞菌标准菌株PAO1,分别在生物膜(静置)和浮游菌(摇床)状态下培养,收集上清液,检测总蛋白酶、LasA和LasB弹性蛋白酶、鼠李糖脂、绿脓素、溶血活性;通过荧光定量PCR检测群体感应(quorum sensing, QS)系统相关基因的表达;同时,通过活菌计数检测PAO1在生物膜和浮游菌状态下的生长曲线。结果生物膜状态下,铜绿假单胞菌PAO1的总蛋白酶、LasA、LasB弹性蛋白酶、鼠李糖脂、绿脓素表达均增高(均P0.05),溶血活性增高(P0.05),生物膜和浮游菌状态下细菌生长曲线差异无统计学意义,QS相关基因rhlI、rhlR、rhlA、lasI、lasR、pqsA、pqsR表达增高(均P0.05)。结论铜绿假单胞菌PAO1在生物膜状态下毒力因子表达较浮游菌状态下增高。     

Evaluation of a one-way airflow system in an animal room based on counts of airborne dust particles and bacteria and measurements of ammonia levels

Air cleanliness in the working area of an animal room equipped with a conventional turbulent flow air distribution systems was compared with that in a similar room fitted with a one-way-flow air distribution system; in this, the supply air flowed from the working area through the racks of cages and was removed from the exhaust side. Before the introduction of animals, the air in the working and exhaust areas of both rooms was ascertained to be Class 100. With animals in situ, however, whereas in the turbulent airflow room both the workspace and exhaust air reached about Class 10,000 (with particle counts, bacterial counts and ammonia levels being almost the same) in the one-way-flow room, the air in the work space only went up to about Class 1000. With the addition of sliding doors or curtains in front of the rack in the one-way-flow room the work space air was maintained at Class 100 with almost no dust particles over 1 microns in size, airborne bacteria or ammonia being detectable. A comparison of all factors measured showed that whereas in the turbulent flow room the contamination of the work space air was 91% of that of the exhaust air, in the one-way-flow room it was only 47%, with curtains added this was reduced to 7% and with sliding doors to only 2%. In the latter case, contamination levels increased markedly on both sides during and immediately after cage changing, but recovered to the pre-cage changing levels within 30 min in the personnel working area and within 60 min on the exhaust side.     

Observation on the air-borne bacteria and ammonia (NS3) gas in laboratory animal facility with rotary heat exchanger]

The number of air-borne bacteria in air ducts and barrierred laboratory animal rooms with the so-called econovent rotary heat exchanger, were checked monthly during a year by the pin-hole sumpler method for air ducts and Koch method for animal rooms. Also, concentration of ammonia was checked with the same samples by gas impinger. No significantly difference in number of air-borne bacteria was seen between before and after passing the econovent. Those passing through HEPA filter was not detected. There were more air-borne bacteria in animal rooms, outside locker room and shower room than in the corridor, utensil storage, inside locker room and pass box. No ammonia were detected in the outdoor, but exhaust air duct after passing the econovent contained very small amount of ammonia. On the other hand, high concentration of ammonia were preserved in the supplying air duct, exhaust air duct and mice and rats rooms, about 86% of ammonia in exhaust air duct returned back into the supplying air duct. No influences on reproduction in mice and rats were recognized.     

The effect of surface charge, negative and bipolar ionization on the deposition of airborne bacteria

Aims:  A series of experiments were conducted to evaluate the effect of surface charge and air ionization on the deposition of airborne bacteria.
Methods and Results:  The interaction between surface electrostatic potential and the deposition of airborne bacteria in an indoor environment was investigated using settle plates charged with electric potentials of 0, ±2·5kV and ±5kV. Results showed that bacterial deposition on the plates increased proportionally with increased potential to over twice the gravitational sedimentation rate at +5kV. Experiments were repeated under similar conditions in the presence of either negative or bipolar air ionization. Bipolar air ionization resulted in reduction of bacterial deposition onto the charged surfaces to levels nearly equal to gravitational sedimentation. In contrast, diffusion charging appears to have occurred during negative air ionization, resulting in an even greater deposition onto the oppositely charged surface than observed without ionization.
Conclusions:  Static charges on fomitic surfaces may attract bacteria resulting in deposition in excess of that expected by gravitational sedimentation or simple diffusion. Implementation of bipolar ionization may result in reduction of bacterial deposition.
Significance and Impact of Study:  Fomitic surfaces are important vehicles for the transmission of infectious organisms. This study has demonstrated a simple strategy for minimizing charge related deposition of bacteria on surfaces.     

Epidemiological investigation of an animal house based upon phage-typing and biotyping of Staphylococcus epidermidis.

The value of biotyping and phage-typing coagulase-negative staphylococci in the epidemiological investigation of a laboratory animal house was clearly demonstrated. In the animal rooms in which conventional bacteriological methods revealed equal bacterial contamination between a conventional unit and one housing specified-pathogen-free rodents, biotyping identified Staphylococcus cohnii as the only species in the latter, compared to S. warneri, S. hominis, S. saprophyticus. S. xylosus abd S. epidermidis as well as S. cohnii in the conventional unit. Similarly, phagetyping revealed 2 phage types in the specified-pathogen-free compared to 7 in the conventional unit. Thus biotyping and phage-typing provided evidence for the existence of a barrier between these units that had presented similar gross bacteriological findings.     

大气污染对大鼠呼吸道微生态的影响研究

探讨了大气污染引起呼吸道症状和机体健康效应的作用机理,为防治大气污染及其对机体的损害提供理论依据。采用模拟现实大气污染的方法,分高、中、低浓度用Wistar大鼠制备大气污染动物模型,对大鼠口咽部菌群进行定性、定量、定位检测,动态研究。大气污染大鼠模型口咽部微生态发生改变。与污染前比,肺炎克雷伯杆菌等致病菌检出率明显增加(P<0.05),厌氧菌含量明显减少(P<0.05),并且随着染毒时间厌氧菌和细菌总量都呈明显下降趋势(P<0.05);口咽部菌群含量在一定范围内波动,没有随着染毒浓度改变而呈明显改变。大气污染对呼吸道正常菌群有毒害作用,对微环境有破坏作用,导致微环境改变,正常菌群含量减少,致病菌定植,是呼吸道疾病发病率升高的重要因素之一。     

四种独立通风笼具（IVC）的检测

目的比较4种独立通风笼具（individually ventilated cages,IVC）的环境参数。方法参考GB14925-2001的检测方法对分别来自4个厂家生产的4种不同的IVC进行环境检测。结果4种IVC在空气洁净度、落下菌、噪声和照度等4个指标检测数据较为一致;在梯度压差、换气次数、气流速度等3个指标存在较大的差别;而温湿度等指标取决于IVC所处的外部环境。结论目前缺乏国家标准对IVC环境指标进行限定,各厂家所生产的IVC存在差异,需要在此方面加强研究,以促进IVC的标准化,更好的服务于实验动物行业。     

The distribution and prevalence of sponges in relation to environmental gradients within a temperate sea lough: vertical cliff surfaces

Abstract. The prevalence and distribution of sponges was surveyed on vertical cliff surfaces at Lough Hyne Marine Nature Reserve, Co. Cork, Ireland. The number of sponge species was recorded at 6-metre depth intervals at four sites within Lough Hyne, and at one site on the adjacent Atlantic coastline to examine differences in abundance and zonation patterns. Sites ranged from an exposed turbulent regime to sheltered, sedimented environments. Individual species showed different distributions and prevalence between sites and with increasing depth. Greatest differences were observed between the most- and least-disturbed sites. Distinct sponge zonation patterns were evident at all sites sampled. Twenty-five species were considered dominant at all five sites with the remaining 48 species considered rare. Only four of the 25 most-dominant species occurred at the site experiencing the most turbulent flow conditions, whereas 12 species were found at the site of unidirectional fast flow. At sites of moderate to slight water movement and high sedimentation, between 18 and 24 of the most dominant species were present. Encrusting forms constituted high proportions of sponge communities at all five sites sampled (although consisting of different species). At sites of turbulent and unidirectional fast flow massive forms also dominated whereas at the least turbulent sites, where sedimentation was high, arborescent sponges were abundant. Few species showed exclusive distribution to a single depth and site, but there was some degree of correlation between species distributions and abiotic factors such as sedimentation rate and flow regimes. Sponge distributions and densities are discussed with respect to the suitability of species' morphologies to particular environments, intra-specific and inter-specific competition and physiological adaptations that enable them to survive in different habitats.     

The role of iron-regulated genes in microbial pathogenesis

An important component of nonspecific defence of human or animal organisms against microbial invasion is attempt to withhold growth-essential iron from invading bacteria. To overcome the barrier bacterial iron uptake systems is derepressed at low iron concentrations. In many pathogenic and nonpathogenic bacteria the regulation of the corresponding genes depends on the regulatory protein Fur. For cloning Fur-regulated loci, methods based on affinity of Fur to the corresponding promoter sites are used.     

Airborne contaminants in conventional laboratory rabbit rooms

Besides the well known allergens, several other risk factors may exist for health in a laboratory animal unit. The exposure to these factors may be significant in animal units with poor general or local ventilation systems. Moreover, means to prevent the distribution of airborne contaminants may be limited in animal units housing rabbits or other bigger laboratory animals. Airborne contaminants in conventional laboratory rabbit rooms were sought to evaluate the occupational exposure of animal care personnel. Concentrations of airborne dust, bacteria, fungi, ammonia and endotoxins were measured during 2 days in three phases: before working activities began, during them and afterwards. Both stationary and some personal samples were taken. All of the contaminants sought were found in the rabbit room air. When compared to reported levels in farm animal production areas, the concentrations measured were generally low. However, moderate or high levels of airborne bacteria and fungi were found occasionally during work routines. Airborne contaminants should be considered as a potential occupational health risk for persons working with laboratory animals.