禽流感病毒实验检测研究进展

禽流感病毒(avian influenza viruses,AIV)给人类健康已带来严重威胁,而实验室快速、准确的诊断技术对禽流感的预防、控制及应急反应决策起着极其关键的作用,这使其成为了研究热点并取得了巨大进步。就禽流感的实验检测技术研究进展从病毒分离、免疫学诊断及分子诊断3个方面加以综述。     

Influence of storage temperature on infectious hematopoietic necrosis virus detection by cell culture isolation and RT-PCR methods

The detection of infectious hematopoietic necrosis virus (IHNV) in infected rainbow trout Oncorhynchus mykiss and in cell culture supernatants stored under different conditions was studied. IHNV-positive fish visceral organ homogenates and cell culture supernatants were incubated at 4 and 25 degrees C. Virus titre was measured by virus isolation on epithelioma papulosum cyprini (EPC) cells and the IHNV RNA was detected by RT-PCR and semi-nested RT-PCR. The influence of repeated freezing and thawing on the virus isolation from organ homogenates and from cell culture supernatants was studied as well. It was possible to isolate the virus from IHNV-positive organ material during the 3 d of incubation at 4 degrees C but, only on the first day of incubation at 25 degrees C. Viral RNA could be amplified during the incubation period of 35 d at 4 degrees C but only during 8 d of incubation at 25 degrees C. In IHNV-infected cell culture supernatant stored at 4 degrees C, it was possible to detect virus for 36 and 16 d in supernatant stored at 25 degrees C. Viral RNA could be followed by using molecular methods during the entire experimental period of 123 d. Each cycle of freezing and thawing of samples resulted in a reduction of IHNV titre in the suspension of visceral organs, while the virus titre in cell culture supernatant remained almost the same following 33 freezing-thawing cycles. The present results show that rapid laboratory processing and storage of potentially virus-containing tissue samples as well as the use of different detection methods are very important for efficient IHNV diagnosis.     

Stability of viruses in waste water sludge eluates

The survival of indigenous enteric viruses in samples of unconcentrated and concentrated waste water sludge eluates, which had been prepared using a combination beef extract elution - organic flocculation concentration procedure, was studied at 2, 23, and -70 degrees C. Changes of virus titer occurring in the samples were followed during an 84-day observation period, with rates of change then calculated by least-squares regression. Virus survival in both types of eluates was statistically dependent (p less than or equal to 0.05) upon storage temperature. Based upon the observed rates of inactivation the average times which would be required for a 90% decrease (one log10 unit) in virus titer for unconcentrated eluates are 27 days at 23 degrees C, 198 days at 2 degrees C, and 375 days at -70 degrees C. The calculated average times required for a 90% decrease in virus titer for concentrated eluates are 22 days at 23 degrees C, 132 days at 2 degrees C, and 246 days at -70 degrees C. In both types of eluates the rates of virus inactivation at 2 degrees C were statistically different from those observed at 23 degrees C, but not different from those observed at -70 degrees C. The three study temperatures were selected to approximate holding of samples in an air-conditioned room, fluid on wet ice (H2O), and frozen on dry ice (CO2).     

Purification and partial characterization of a thermostable trithionate hydrolase from the acidophilic sulphur oxidizer Thiobacillus acidophilus.

Cell-free extracts of Thiobacillus acidophilus catalysed the quantitative conversion of trithionate (S3O6(2-) to thiosulphate and sulphate. A continuous assay for quantification of experimental results was based on the difference in absorbance between trithionate and thiosulphate at 220 nm. Trithionate hydrolase was purified to near homogeneity from cell-free extracts of T. acidophilus. The molecular masses of the native enzyme and the subunit were 99 kDa (gel filtration) and 34 kDa (SDS/PAGE). The purified enzyme has a pH optimum of 3.5-4.5 and a temperature optimum of 70 degrees C. Enzyme activity was stimulated by sulphate. The stimulation of the enzyme activity by sulphate was half maximal at a concentration of 0.23 M. The Km for trithionate is 70 microM at 30 degrees C and 270 microM at 70 degrees C. Enzyme activity was lost after 36 days at 0 degrees C, 27 days at 70 degrees C; but after 97 days at 30 degrees C, 40% of the initial activity was still present: The enzyme activity was inhibited by mercury chloride, N-ethylmaleimide, thiosulphate and tetrathionate. Tetrathionate S4O6(2-) was not hydrolysed by trithionate hydrolase.     

Persistence of the 2009 pandemic influenza A (H1N1) virus in water and on non-porous surface

Knowledge of influenza A virus survival in different environmental conditions is a key element for the implementation of hygiene and personal protection measures by health authorities. As it is dependent on virus isolates even within the same subtype, we studied the survival of the 2009 H1N1 pandemic (H1N1pdm) virus in water and on non-porous surface. The H1N1pdm virus was subjected to various environmental parameters over time and tested for infectivity. In water, at low and medium salinity levels and 4°C, virus survived at least 200 days. Increasing temperature and salinity had a strong negative effect on the survival of the virus which remained infectious no more than 1 day at 35°C and 270 parts per thousand (ppt) of salt. Based on modeled data, the H1N1pdm virus retained its infectivity on smooth non-porous surface for at least 7 days at 35°C and up to 66 days at 4°C. The H1N1pdm virus has thus the ability to persist in water and on glass surface for extended periods of time, even at 35°C. Additional experiments suggest that external viral structures in direct contact with the environment are mostly involved in loss of virus infectivity.     

Growth of epizootic hemorrhagic disease, akabane, and ephemeral fever viruses in Aedes albopictus cells maintained at various temperatures

The growth curves of one epizootic hemorrhagic disease (EHD) virus serotype (Reoviridae), two Akabane virus strains (Bunyaviridae) and three bovine ephemeral fever (BEF) group viruses (Rhabdoviridae) were determined in Aedes albopictus cells maintained at 15, 20, 28 and 33 degrees C. Ae albopictus cells supported the growth of all the viruses although not necessarily at all temperatures. Because none of the viruses exhibited cytopathic effect in Ae albopictus cells, growth was assayed in baby hamster kidney 21 (BHK21) cells maintained at 37 degrees C. The temperature at which the Ae albopictus cells were maintained had a marked effect on the growth and yield for each virus studied. EHD virus was heat-stable and grew after 4 days at 28 and 33 degrees C, and after 8 days at 20 degrees C. No growth was recorded up to 12 days at 15 degrees C. The two Akabane viruses were heat-sensitive and exhibited different growth patterns. One strain (B8935) showed no growth at 15 degrees C and only minimal growth at 20, 28 and 33 degrees C. The other strain (CSIRO 16) showed growth after 1-2 days at all temperatures with higher titres reached at 15 and 20 degrees C than at 28 and 33 degrees C. The BEF group viruses grew to approximately the same titres at all temperatures. At the higher temperatures (28 and 33 degrees C) most of BEF group viruses had disappeared within 9 days. In contrast at the lower temperatures (15 and 20 degrees C), there was still virus present 18 days after inoculation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Rotavirus survival in conventionally treated drinking water

Samples of conventionally treated drinking water collected either as effluent (PE) at a treatment plant or out of a tap (TW) in our laboratory were seeded with simian rotavirus SA-11, which closely resembles rotavirus of human origin. The virus, grown in MA-104 cells, was suspended either in distilled water, Earle's balanced salt solution (EBSS), or tryptose phosphate broth (TPB), and added to the water samples to a final concentration of 5.7 X 10(3) plaque-forming units (PFU) per millilitre. After a contact time of 1 h at 22 degrees C, the samples were diluted and plaque assayed. There was no significant reduction in the virus titre in samples of TW (less than 0.05 mg/L free chlorine). The titre also remained almost the same in PE (0.75 mg/L free chlorine) when EBSS or TPB was used for virus suspension. There was, however, nearly a 1 log10 loss in the titre of the virus when it was suspended in distilled water before the contamination of PE. To study the long-term survival of the rotavirus in TW, the inoculated samples (5.0 X 10(4) PFU/mL) were held at either 4 or 20 degrees C in the dark and tested over a period of 64 days. At 20 degrees C it took 64 days to reduce the virus titre by 2 log10, whereas at 4 degrees C the virus titre dropped only 0.7 log10 during the same period. Rotaviruses could, therefore, survive well enough in conventionally treated drinking water to make it a possible vehicle for their transmission.     

Survival of spumavirus,a primate retrovirus,in laboratory media and water

The persistence of a previously characterized spumavirus strain (strain SV-522) was investigated utilizing various laboratory media and waters, including Eagle's minimal essential medium (EMEM) plus 0% fetal bovine serum (EMEM-0%), EMEM-2%, EMEM-10%, Chlamydia transport medium (CTM), phosphate-buffered saline, distilled, estuarine, and marine water, human serum, and the germicides, ethyl alcohol (70%) and sodium hypochlorite (10%). Experiments were performed at 4 degrees C and/or 23 degrees C. Infectivity endpoints were determined in stock aliquots upon initiation of testing and then after 3, 5, 7, and 10 days. The virus was reisolated from all diluents after 5 days at 23 degrees C and in EMEM-10% after 7 days. The virus was detected in CTM, EMEM-2%, EMEM-10%, and estuarine and marine waters after 7 days at 4 degrees C. Differences in the persistence of the virus may be ascribed to temperature and organic load. Water ionic strengths (e.g., estuarine vs. marine water) had no effect on modifying persistence of viral particles. Infectivity of spumavirus was undetectable after 30 s in 70% ethanol or 10% sodium hypochlorite. After 30 min at 23 degrees C, spumavirus infectivity in normal but not heat-inactivated human serum increased by almost 100-fold. Persistence of infectivity of primate spumavirus after 7 days in media and waters, and the agent's infectious potential in the human host, emphasize a need for cautious recognition during the manipulation of primate cells/organs and in the handling of primates themselves.     

Long-term survival of human rotavirus in raw and treated river water

This study was aimed at assessing the role of water as a vehicle for rotavirus spread by determining how well these viruses survive in the water environment. A cell culture adapted strain of human rotavirus subgroup 2, grown in MA-104 cells, was used as a model. Virus survival was tested in the following types of water samples, derived from the Ottawa River, at two different times of the year: (i) raw water (RW), (ii) muncipally treated tap water (TW), and (iii) raw water that had been filtered (FW) through a membrane (0.22 micron). The water samples, with approximately 5.0 X 10(4) plaque-forming units (PFU) of the virus, were held at either 4 or 20 degrees C and tested for infectious virus over a period of 64 days. The TW samples had a total and free chlorine content of 0.05 and less than 0.05 mg/L, respectively. The chlorine in these samples was not neutralized before virus contamination. Irrespective of the holding temperature, the virus titre in FW remained essentially unaltered throughout the test period. In TW held at 4 degrees C, there was no significant drop in the virus titre even after 64 days, whereas at 20 degrees C the titre in TW was reduced by about 2 log10 over the same period. Even though the loss of virus infectivity was most rapid in RW held at 20 degrees C, it took about 10 days for a 99.0% reduction in the plaque titre of the virus. These findings, therefore, indicate that rotaviruses can survive for several days in raw and treated river water thus making recreational and potable waters potential vehicles for the transmission of rotavirus infections.     

Evaluation of the genetic stability of the temperature-sensitive PB2 gene mutation of the influenza A/Ann Arbor/6/60 cold-adapted vaccine virus.

A single-gene reassortant bearing the PB2 gene of the A/Ann Arbor/6/60 cold-adapted virus in the background of the A/Korea/82 (H3N2) wild-type virus is a temperature-sensitive (ts) virus with an in vitro shutoff temperature of 38 degrees C. A single mutation at amino acid (aa) at 265 (Asp-Ser) of the PB2 protein is responsible for the ts phenotype. This ts single-gene PB2 reassortant virus was serially passaged at elevated temperatures in Madin-Darby canine kidney cells to generate ts+ phenotypic revertant viruses. Four ts+ phenotypically revertant viruses were derived independently, and each possessed a shutoff temperature for replication in vitro of > 40 degrees C. Each of the four phenotypically revertant viruses replicated efficiently in the upper and lower respiratory tracts of mice and hamsters, unlike the PB2 single-gene reassortant virus, confirming that the ts phenotype was responsible for the attenuation of this virus in rodents. Mating the ts+ revertants with wild-type virus yielded ts progeny in high frequency, indicating that the loss of ts phenotype was due to a suppressor mutation which was mapped to the PA gene in each of the four independently derived ts phenotypic revertants. Nucleotide sequence analysis confirmed the absence of new mutations on the PB2 gene and the presence of predicted amino acid changes in the PA proteins of the revertant viruses. These studies suggest that single amino acid changes at aa 245 (Glu-Lys) or 347 (Asp-Asn) of the PA protein can completely suppress the ts and attenuation phenotypes specified by the Asp-Ser mutation at aa 265 of the PB2 protein of the A/Ann Arbor/6/60 cold-adapted virus.     

Long-term survival of hepatitis A virus and poliovirus type 1 in mineral water.

The survival in mineral water of hepatitis A virus (HAV) and poliovirus type 1 was compared, under controlled experimental conditions, at 4 degrees C and room temperature. Viral infectivity titers were determined by cell culture titration, while HAV antigenicity was monitored by radioimmunoassay-endpoint titration. Both viruses persisted longest at 4 degrees C. At this temperature, after 1 year of exposure, the inactivation of either HAV or poliovirus type 1 was not important. At room temperature, poliovirus type 1 was not detected after 300 days, whereas HAV was still infectious. For both temperatures, the computed regression coefficients of best-fit lines for inactivation rates for the two viruses were significantly different. The survival of HAV was also studied at 4 degrees C and room temperature in mineral water with 5- and 50-micrograms/ml protein concentrations (i.e., purity of the virus suspension) for 120 days. As shown by a comparison of the regression coefficients for the inactivation rates, the stability of HAV in mineral water depends on protein concentration and temperature. Radioimmunoassay-endpoint titration results showed inactivation patterns similar to those of cell culture titration, with the most significant reduction in HAV antigenicity at room temperature. At the two temperatures, the infectivity of HAV declined at a faster rate than the antigenicity.     

土壤温度和水分对长白山不同森林类型土壤呼吸的影响

在实验室条件下,将不同含水量的3种森林类型的土柱分别置于0、5、15、25和35℃条件下,进行土壤呼吸测定．结果表明,在0～35℃范围内。土壤呼吸速率与温度呈正相关．在一定含水量范围内(0．21～0．37kg·kg^-1),土壤呼吸随含水量的增加而升高,当含水量超出该范围,土壤呼吸速率则随含水量的变化而降低．土壤温度和水分对土壤呼吸作用存在明显的交互作用．不同森林类型土壤呼吸作用强弱存在显著差异,大小顺序为阔叶红松林>岳桦林>云冷杉暗针叶林．阔叶红松林土壤呼吸作用的最佳条件是土壤温度35℃、含水量0．37kg·kg^-1;云冷杉暗针叶林下的山地棕色针叶林土壤呼吸作用的最佳条件是25℃、0．21kg·kg^-1;岳桦林土壤呼吸作用的最佳条件是35℃、含水量0．37kg·kg^-1。但是．由于长白山阔叶红松林、云冷杉林和岳桦林处在不同的海拔带上,同期不同森林类型土壤温度各不相同,相差4～5℃,所以野外所测的同期山地棕色针叶林土呼吸速率应低于暗棕色森林土呼吸速率,山地生草森林土呼吸速率应高于山地棕色针叶林土的呼吸速率．     

Control of local heat gain by vasomotor response of the hand

Finger blood flow (BF) was measured by venous occlusion plethysmography using mercury-in-Silastic strain gauges during immersion of one hand in hot water (raised by steps of 2 degrees C every 10 min from 35 to 43 degrees C), the other being a control (kept immersed in water at 35 degrees C). The measurements were made in three different thermal states on separate days: 1) cool-25 degrees C, 40% rh, esophageal temperature (Tes) = 36.64 +/- 0.10 degrees C; 2) warm-35 degrees C, 40% rh, Tes = 36.71 +/- 0.11 degrees C; and 3) hot-35 degrees C, 80% rh with the legs immersed in water at 42 degrees C, Tes = 37.26 +/- 0.11 degrees C. When water temperature was raised at 42 degrees C, Tes = 37.26 +/- 0.11 When water temperature was raised to 39-41 degrees C in the warm state, finger BF in the hand heated locally (BFw) decreased. When water temperature was raised to 43 degrees C, however, BFw returned to the control value. In the hot state, Tes rose steadily, reaching 37.90 +/- 0.12 degrees C at the end of the 50-min sessions. BF in the control finger also increased gradually during the session. BFw showed a tendency to decrease when water temperature was raised to 39 degrees C, but the change was not greater than that observed in the warm state. In the cool state, no such reduction in BFw was observed when water temperature was raised to 39-41 degrees C. On the contrary, BFw increased at water temperatures of 41-43 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of incubation temperature on in vitro maturation of porcine oocytes: nuclear maturation, fertilisation and developmental competence.

The present study examined the effect of low culture temperature during in vitro maturation (IVM) of pig oocytes on their nuclear maturation, fertilisation and subsequent embryo development. In experiment 1, oocytes were cultured at 35 or 39 degrees C for 44 h in modified tissue culture medium 199 supplemented with 10 ng/ml epidermal growth factor, 0.57 mM cysteine, 75 microg/ml potassium penicillin G, 50 microg/ml streptomycin sulphate, 0.5 microg/ml LH and 0.5 microg/ml FSH to examine the nuclear maturation status. In experiment 2, oocytes were cultured at 35 degrees C for 44 or 68 h and nuclear maturation was examined. In experiment 3, oocytes matured for 44 or 68 h at 39 degrees C and for 68 h at 35 degrees C were co-incubated with frozen-thawed spermatozoa for 5-6 h. Putative embryos were transferred into North Carolina State University (NCSU) 23 medium containing 0.4% bovine serum albumin. At 12 h after insemination, some oocytes were fixed to examine the fertilisation rate and the remaining embryos were examined at 48 and 144 h for cleavage and blastocyst formation rate, respectively. Compared with 39 degrees C, culture of oocytes at 35 degrees C for 44 h significantly (p < 0.05) reduced the metaphase II (M II) rate (79% vs 12%). However, extension of culture time to 68 h at 35 degrees C significantly increased (p < 0.05) the M II rate (7% vs 58%). In experiment 3, compared with other groups, fewer (p < 0.05) oocytes reached M II when cultured at 35 degrees C for 68 h (69-81% vs 49%). Extension of culture duration to 68 h at 39 degrees C stimulated spontaneous activation (28%) of oocytes. No difference in cleavage rates was observed among different groups. Compared with oocytes matured for 44 h at 39 degrees C (31%), the proportion of blastocysts obtained was low (p < 0.05) for oocytes matured at 35 degrees C (13%) or 39 degrees C (3%) for 68 h. The results indicate that lower culture temperature can delay nuclear maturation of pig oocytes. However, extension of culture time can stimulate nuclear maturation and these oocytes are capable of fertilisation and development to the blastocyst stage at moderate rates.     

Effect of water temperature on the immune response and infectivity pattern of white spot syndrome virus (WSSV) in freshwater crayfish

The susceptibility of two species of freshwater crayfish, Pacifastacus leniusculus and Astacus astacus, to white spot syndrome virus (WSSV) by intramuscular injection was compared and the results show that both species are susceptible to WSSV. The effect of water temperature on the development of white spot disease in crayfish was also studied. Crayfish were exposed to different temperatures after WSSV injection or oral exposure and the mortalities were recorded over a period of 45 days. No mortality was observed when crayfish were held at 4+/-2 degrees C or 12+/-2 degrees C and reached 100% when these crayfish transferred to 22+/-2 degrees C. The mortalities of nearly moribund crayfish at 22+/-2 degrees C with WSSV could be delayed after transfer to temperature below 16 degrees C. These results clearly show that low temperature affects the WSSV pathogenicity in crayfish. Moreover, haemocyte counts, phenoloxidase activity, mRNA levels of prophenoloxidase (proPO) and the lipopolysaccharide and beta-1,3-glucan binding protein (LGBP) in crayfish exposed to various water temperatures were studied. Total haemocyte and granular cell counts of crayfish held at different temperatures were not significantly (P>0.05) different, except for the total haemocyte number at 18 degrees C was significantly (P<0.05) higher than in crayfish at 4 degrees C. The percentage of granular cells in crayfish held at 4 degrees C was the highest compared to crayfish maintained at other temperatures. The phenoloxidase activities in haemocyte lysate supernatant (HLS) of crayfish at all temperature groups remained similar. The amount of proPO-mRNAs in haemocytes was much higher than the amount of LGBP-m RNAs in all the experimental groups. However, there was no change in the level of pro PO-mRNA at the tested temperatures. Interestingly, the level of LGBP-mRNA of crayfish kept at 22 degrees C was much lower than in those held at lower temperatures. Proliferation of the haematopoietic tissues was higher at high temperatures which may support replication of WSSV, and explain the high mortality of crayfish with WSSV infection at high temperature. Based on these studies it is concluded that crayfish might act as a carrier of WSSV at low water temperature and could develop white spot disease if the water temperature is increased.     

Life history of hawthorn spider mite Amphitetranychus viennensis (Acarina: Tetranychidae) on various apple cultivars and at different temperatures

Development duration and reproduction rate of hawthorn spider mite Amphitetranychus viennensis (Zacher) were carried out on five different apple cultivars (Amasya (local cultivar), Golden Delicious, Granny Smith, Starking Delicious and Starkrimson Delicious) at 25 degrees C, 65 +/- 10% RH and 16:8 L:D. In addition, the same parameters were determined on Golden Delicious leaves at three constant temperatures (20, 30 and 35 degrees C, 65 +/- 10% RH and 16:8 L:D) in the laboratory. A. viennensis showed a better performance on Golden Delicious than on the other apple cultivars. This was mainly due to a short development time (10.7 days), high daily egg production (5.2 eggs/female/day) and early reproduction peak. The highest intrinsic rate of natural increase (rm) was determined on the variety Golden Delicious (rm = 0.247/day), while the lowest one was observed on the variety Starking Delicious (rm = 0.215/day). The developmental periods of A. viennensis varied from 7.4 to 18.8 days at 35 and 20 degrees C for females, while it varied from 7.9 to 17.2 days at 30 and 20 degrees C for males. The development threshold of the eggs and pre-adult stages were 9.72 and 9.07 degrees C, total effective temperature was 72.99 and 185.18 degree-days, respectively. The mean generation time (To) of the population ranged from 16.13 days at 30 degrees C to 29.15 days at 20 degrees C. The net reproductive rate (R0) increased from 54.33 female/female at 20 degrees C to 78.34 female/female at 25 degrees C, and decreased to 75.71 female/female at 30 degrees C. The highest intrinsic rate of increase (rm) was reached at 30 C (rm = 0.268/day), the lowest one at 20 degrees C (rm = 0.136/day).     

Survival of human parainfluenza viruses in the South Polar environment

The survival of human parainfluenza virus types 1, 2, and 3 was measured in both indoor and outdoor environments at South Pole Station, Antarctica, in an effort to determine the long-term survival of these viruses in this environment and to identify the possible source of respiratory tract illnesses which occurred in this isolated population in 1978 after 10 and 27 weeks of total social isolation. Viruses were applied to plastic petri plate surfaces which were then stored in indoor (21.4 degrees C; water vapor density, 1.50 g of water per m3) and outdoor environments (-22.4 to -33.2 degrees C; water vapor density, 0.706 and 0.247 g of water per m3). Parainfluenza virus type 1 at an initial titer of 3.75 log10 50% tissue culture infective doses per ml was inactivated after 4 days at room temperature and after 7 days outside. Parainfluenza virus type 2 and 3 at initial titers of 5.58 and 5.38 log10 50% tissue culture infective doses per ml were inactivated after 7 and 12 days, respectively, at room temperature and after 17 days of storage outside. Results indicate that the long-term survival of parainfluenza virus in either environment for up to 10 weeks is unlikely and probably did not provide the source of infectious virus responsible for the midisolation outbreaks of parainfluenza virus-related respiratory tract illnesses observed in this population during the 1978 winter season.     

Survival of human parainfluenza viruses in the South Polar environment.

The survival of human parainfluenza virus types 1, 2, and 3 was measured in both indoor and outdoor environments at South Pole Station, Antarctica, in an effort to determine the long-term survival of these viruses in this environment and to identify the possible source of respiratory tract illnesses which occurred in this isolated population in 1978 after 10 and 27 weeks of total social isolation. Viruses were applied to plastic petri plate surfaces which were then stored in indoor (21.4 degrees C; water vapor density, 1.50 g of water per m3) and outdoor environments (-22.4 to -33.2 degrees C; water vapor density, 0.706 and 0.247 g of water per m3). Parainfluenza virus type 1 at an initial titer of 3.75 log10 50% tissue culture infective doses per ml was inactivated after 4 days at room temperature and after 7 days outside. Parainfluenza virus type 2 and 3 at initial titers of 5.58 and 5.38 log10 50% tissue culture infective doses per ml were inactivated after 7 and 12 days, respectively, at room temperature and after 17 days of storage outside. Results indicate that the long-term survival of parainfluenza virus in either environment for up to 10 weeks is unlikely and probably did not provide the source of infectious virus responsible for the midisolation outbreaks of parainfluenza virus-related respiratory tract illnesses observed in this population during the 1978 winter season.     

Partitional measurement of capillary and arteriovenous anastomotic blood flow in the human finger by laser-Doppler-flowmeter

This study was made to see whether changes in blood flow through the capillaries and arteriovenous anastomoses (AVA's) of the human finger can be measured by noninvasive flowmetry. Total finger blood flow (FBF) was measured by venous occlusion plethysmography; blood flow was measured by a laser-Doppler flowmeter (ADVANCE, ALF-2100, Tokyo, Japan) using probes with optic fiber separations of 0.3 mm (LDF-0.3) and 0.7 mm (LDF-0.7). The maximum sensitivities for LDF-0.3 and LDF-0.7 were at depths of 0.8 and 1.2 mm from the tissue surface respectively. Two series of experiments were performed on separate days. In the first series the test hand was immersed in a water bath whose temperature (Tw) was 25 degrees C at an ambient temperature (Ta) of 25 degrees C. Tw was raised to 35 degrees C (local hand warming), which was then followed by an increase in Ta to 35 degrees C (whole body warming). FBF, LDF-0.3, and LDF-0.7 increased during these thermal stimulations. However, the relationship of FBF to LDF-0.3 showed two different regression lines. In contrast, the relationship of FBF to LDF-0.7 showed a single regression line. In the second series, with Ta at 35 degrees C, the test hand was immersed in a water bath at Tw 35 degrees C. Tw was then raised every 10 min by 2 degrees C steps from 35 to 41 degrees C. At Tw 39-41 degrees C, FBF and LDF-0.7 in the test hand were significantly decreased compared with those at Tw 35 degrees C.(ABSTRACT TRUNCATED AT 250 WORDS)