^60Coγ射线对刚地弓形虫缓殖子感染性的影响

应用不同剂量的~(60)Coγ射线照射含NT株弓形虫包囊的鼠脑匀浆后,以0.4％胰蛋白酶液消化,使弓形虫缓殖子从包囊中释放出来,并用生理盐水作连续10倍稀释,使成为含不同数量缓殖子的悬液,将此悬液接种小鼠,用生物检测方法确定其感染性。另设未经照射处理的缓殖子悬液感染小鼠的对照组。结果表明,γ射线0.55kGy剂量为控制鼠脑中NT株弓形虫感染性的最小有效剂量。γ射线0.1kGy剂量对弓形虫缓殖子感染性无明显影响,而0.45kGy照射后,其感染性较前者下降了10~4倍。     

Inactivation of Giardia muris Using Ozone and Ozone-Hydrogen Peroxide

The disinfection effects of the ozone molecule alone and that of ozone decomposition products when inactivating Giardia muris cysts were investigated at bench-scale using two different ozone demand-free laboratory buffer systems. The first water was a 0.05 M phosphate buffer with hydrogen peroxide added at a 10:1 weight ratio. The second water was a 0.05 M phosphate – 0.01 M bicarbonate buffer which quickly scavenged radical species from ozone decomposition. The C3H/HeN mouse model was used to assess the infectivity of ozone treated cysts.

The phosphate-bicarbonate buffer system had significantly greater (P ≤ 0.05) inactivation of G. muris cysts than that observed in the phosphate buffer – peroxide system where ozone was completely decomposed in less than 120 s. Consequently, the design of ozone disinfection processes should maintain ozone residual for disinfection prior to the addition of hydrogen peroxide for the oxidation of other compounds.     

森林脑炎病毒不同感染途径对小鼠模型的感染性分析

目的分析森林脑炎病毒不同感染途径对小鼠模型的致病性、病毒分布及增殖动态。方法将适应原代地鼠肾(primary hamster kidney,PHK)细胞的森林脑炎病毒进行10倍系列稀释,取4个连续稀释度的病毒液,分别通过脑腔注射、灌胃、滴鼻、肌肉注射、腹腔注射的感染方式,对昆明小鼠进行攻毒,逐日连续观察14 d,计算半数致死量(median lethal dose,LD50);并分别取攻毒后第3、5、7天发病典型的腹腔感染组小鼠脑、肺、肾脏、肝脏、心脏、小肠、血液7种组织器官,采用蚀斑法检测病毒滴度。结果脑腔、灌胃、滴鼻、肌肉注射和腹腔注射攻毒方式的LD50分别为10、105.1、105、101.2和102PFU/mL。所有攻毒途径小鼠的脑腔中病毒滴度最高,达109PFU/mL,肺组织为106.5PFU/mL,肾脏为105PFU/mL,心脏和肝脏中病毒滴度较低,分别为102和101.5PFU/mL,肠道中出现一过性病毒感染,而血液中病毒滴度最高为105.5PFU/mL。结论森林脑炎病毒可通过多种途径感染小鼠模型,除具有较强易感性的脑组织外,对肺组织细胞同样具有较强的感染性。     

UV inactivation and resistance of rotavirus evaluated by integrated cell culture and real-time RT-PCR assay

Rotaviruses are double-stranded RNA viruses which are among the most resistant waterborne enteric viruses to UV disinfection. An integrated cell culture and real-time RT-PCR (ICC real-time RT-PCR) assay was developed to detect the infectivity of rotaviruses in water, which uses real-time RT-PCR to detect RNA produced by infectious rotaviruses during replication in host cells. Detection of rotaviral RNA in host cells provides direct evidence of the presence of infectious rotavirus rather than just the presence of rotavirus RNA. Using this newly developed method, the inactivation and resistance of rotavirus to UV treatments at various doses was evaluated. With an initial concentration of 2 × 10⁴ PFU/ml simian rotavirus (SA11), a first-order linear relationship was obtained at UV dose range of 0-120 mJ cm⁻², and the inactivation rate constant was estimated to be 0.0343 cm² mJ⁻¹ (R² = 0.966). The dose-inactivation curve tailed off and reached plateau as the UV dose increased from 120 to 360 mJ cm⁻², indicating resistance phenomena of sub-populations of SA11 at very high UV doses. A maximal reduction of 4.8 log₁₀ was observed. Through parallel comparison with traditional culture assay, the ICC real-time RT-PCR method demonstrated more effective, sensitive and faster infectivity detection of rotavirus and, the results reveal that rotaviruses are more resistant to UV irradiation than previously reported with traditional cell culture assays.     

Infectivity of Giardia lamblia cysts obtained from wastewater treated with ultraviolet light

Several waterborne outbreaks of giardiasis have been linked to discharge of wastewater effluents into surface water. Little is known about the infectivity of Giardia lamblia cysts present in UV treated wastewater effluents. In this study, the infectivity of G. lamblia cysts, recovered from primary effluent and secondary effluent, both upstream and downstream of operating full-scale UV reactors at four wastewater treatment plants, was assessed using the Mongolian gerbil model. Infectivity of cysts obtained from the primary effluents was scored as either strong or moderate for induction of infection in gerbils at three out of four wastewater treatment plants. G. lamblia recovered from secondary effluent both upstream and downstream of the UV reactors caused weak infections in the gerbils. The probability of weak infections caused by inoculums of 50-1400 cysts per gerbil was, on the average, reduced by approximately 10% at the four wastewater UV installations with coliform reduction equivalent doses ranging from 6 to 18 mJ/cm². The UV systems provided considerably less inactivation of the parasite than expected based on the UV dose response of Giardia reported in the literature.     

Synergistic inactivation of Cryptosporidium parvum using ozone followed by monochloramine in two natural waters

The effect of sequential exposure to ozone followed by monochloramine on inactivation of Cryptosporidium parvum oocysts suspended in untreated natural surface water from two different sources was studied in bench-scale batch reactors. Animal infectivity using neonatal CD-1 mice was used to measure oocyst inactivation. A statistically significant synergistic effect on oocyst inactivation was measured in both natural water samples studied. The magnitude of the effect measured in the natural water with lower turbidity, colour, and organic carbon concentration was comparable to that previously reported for oocysts suspended in buffered de-ionized water but was reduced considerably in the natural water with higher turbidity, colour and organic carbon concentration. Synergy increased with initial pH and with the degree of ozone pre-treatment but was independent of temperature. For water treatment plants with adequate disinfectant contact times, ozone followed by monochloramine may be a practical means of achieving additional C. parvum inactivation, however, the influence of water quality characteristics should be considered.     

Changes in culturability and virulence of Salmonella typhimurium during long-term starvation under desiccating conditions

The survival of Salmonella typhimurium under desiccation and starvation conditions commonly associated with farm buildings was investigated in a desiccation model system: filtration onto polycarbonate membranes placed in a sealed desiccator with 0.0067 g/m³ absolute humidity. Heterogeneities within bacterial populations in relation to time of desiccation were investigated on a single-cell basis by epifluorescence microscopy coupled with an image analysis system in conjunction with fluorescent dyes Chemchrome V6 and DAPI. Changes in cellular states were compared to the results of plate counts (colony forming units, CFU) on selective (modified semi-solid Rappaport Vassiliadis (MSRV)) and non-selective (nutrient agar (NA) and R2A agar) media, and to the measurements of infectivity and virulence using two animal models (chicks and mice). During 9 weeks of experimental desiccation, total cell counts (DAPI) of starved S. typhimurium remained stable, as did esterase activity (Chemchrome V6), but DAPI fluorescence intensity decreased slowly. Bacterial cells entered gradually into non-culturable states (decrease of CFU counts on MSRV, NA and R2A agar media) and the total loss of culturability on NA (defined as probability of presence of 1 CFU on the membrane inferior to 10⁻⁶) was obtained after 9 weeks. Loss of chick infectivity and mice virulence in animal models occurred more rapidly, within three weeks of experimental desiccation.     

Changes in culturability and virulence of Salmonella typhimurium during long-term starvation under desiccating conditions

The survival of Salmonella typhimurium under desiccation and starvation conditions commonly associated with farm buildings was investigated in a desiccation model system: filtration onto polycarbonate membranes placed in a sealed desiccator with 0.0067 g/m³ absolute humidity. Heterogeneities within bacterial populations in relation to time of desiccation were investigated on a single-cell basis by epifluorescence microscopy coupled with an image analysis system in conjunction with fluorescent dyes Chemchrome V6 and DAPI. Changes in cellular states were compared to the results of plate counts (colony forming units, CFU) on selective (modified semi-solid Rappaport Vassiliadis (MSRV)) and non-selective (nutrient agar (NA) and R2A agar) media, and to the measurements of infectivity and virulence using two animal models (chicks and mice). During 9 weeks of experimental desiccation, total cell counts (DAPI) of starved S. typhimurium remained stable, as did esterase activity (Chemchrome V6), but DAPI fluorescence intensity decreased slowly. Bacterial cells entered gradually into non-culturable states (decrease of CFU counts on MSRV, NA and R2A agar media) and the total loss of culturability on NA (defined as probability of presence of 1 CFU on the membrane inferior to 10⁻⁶) was obtained after 9 weeks. Loss of chick infectivity and mice virulence in animal models occurred more rapidly, within three weeks of experimental desiccation.     

Biology, persistence and detection of Cryptosporidium parvum and Cryptosporidium hominis oocyst

Cryptosporidium parvum and Cryptosporidium hominis are obligate enteric protozoan parasites which infect the gastrointestinal tract of animals and humans. The mechanism(s) by which these parasites cause gastrointestinal distress in their hosts is not well understood. The risk of waterborne transmission of Cryptosporidium is a serious global issue in drinking water safety. Oocysts from these organisms are extremely robust, prevalent in source water supplies and capable of surviving in the environment for extended periods of time. Resistance to conventional water treatment by chlorination, lack of correlation with biological indicator microorganisms and the absence of adequate methods to detect the presence of infectious oocysts necessitates the development of consistent and effective means of parasite removal from the water supply. Additional research into improving water treatment and sewage treatment practices is needed, particularly in testing the efficiency of ozone in oocyst inactivation. Timely and efficient detection of infectious C. parvum and C. hominis oocysts in environmental samples requires the development of rapid and sensitive techniques for the concentration, purification and detection of these parasites. A major factor confounding proper detection remains the inability to adequately and efficiently concentrate oocysts from environmental samples, while limiting the presence of extraneous materials. Molecular-based techniques are the most promising methods for the sensitive and accurate detection of C. parvum and C. hominis. With the availability of numerous target sequences, RT-PCR will likely emerge as an important method to assess oocyst viability. In addition, a multiplex PCR for the simultaneous detection of C. parvum, C. hominis and other waterborne pathogens such as Giardia lamblia would greatly benefit the water industry and protect human health.