Sporadic Occurrence of Echo Virus Types 27 and 31 Associated with Aseptic Meningitis in Ontario

The sporadic occurrence of Echo virus types 27 and 31 in association with aseptic meningitis during the enterovirus seasons in Ontario in 1959 to 1962 is reported.The etiological significance of both of these isolates was verified by demonstration of type-specific serological response in meningitis patients. Echo 27, which up to now has not been encountered in association with human illness, could be added to the list of viral agents capable of causing aseptic meningitis. Isolation of Echo 31 from cerebrospinal fluid provides definite proof of the etiological significance of this virus in cases of aseptic meningitis.     

Viral Infections of Toronto Children During 1965: I. Enteroviral Disease

Enteroviruses were isolated from feces and/or cerebrospinal fluid of 29 of 43 Toronto children who contracted aseptic meningitis, pleurodynia, abdominal pain or febrile upsets between June and October, 1965. Coxsackie A9 virus was the dominant agent in aseptic meningitis and Coxsackie B1 virus in pleurodynia and other syndromes. Sero-logical evidence of recent Coxsackie B1 and Echo 6 infection was obtained in two additional patients with aseptic meningitis who did not yield virus, and elevated Coxsackie B1 antibody titres were found in one patient with pericarditis. A newborn infant died with myocarditis due to Coxsackie B1 virus following infection of the mother during the immediate antenatal period. Paired sera collected only two to four days apart from patients with enteroviral syndromes or mumps meningoencephalitis frequently showed four-fold or greater increases of antibody levels.     

Enteroviral Syndromes in Toronto, 1964

Virological or serological investigations of 72 children in Toronto and environs, who were hospitalized between January and October 1964 with a variety of syndromes, revealed evidence of enteroviral infection in 29 subjects. Coxsackie B2 was the dominant enterovirus, being isolated from feces and/or cerebrospinal fluid (CSF) of three children with aseptic meningitis, three with pleurodynia, one with myalgia and one with pericarditis; four additional patients showed rising antibody titres to this virus. Coxsackie B1 virus, which has not been isolated in Toronto since 1950, was recovered from feces of three patients with pleurodynia, CSF of one patient with myalgia, and peritoneal fluid of a child with primary peritonitis; one patient with pericarditis showed a rising antibody titre to Coxsackie B1 virus. Coxsackie B3, B4 and Echo 23 viruses were associated with one case each of pleurodynia. Coxsackie B5 virus infected five patients with aseptic meningitis, and one each with pericarditis and myocarditis.     

Mumps and Enteroviral Meningitis in Toronto, 1966

Of 52 children admitted to hospital for apparently typical mumps meningitis in 1966, 50 had their cerebrospinal fluid (CSF) examined. In only 17 was the mumps virus isolated from the CSF. Mumps antihemagglutinin conversions or increments were detected in 32 subjects including 10 whose CSF yielded virus. Antibody conversions were found in 16 patients and fourfold increments in another nine whose serum pairs were collected only one to four days apart. Initial sera from 20 patients were obtained three days or less after the onset of meningitis. Antibody increments were frequently noted about one day after defervescence and clinical improvement. Interferon was detected in CSF from two of eight patients, both of whom yielded virus. Enteroviruses were isolated from CSF and/or feces in seven of 15 cases of aseptic meningitis which occurred between July and October. Six patients including three virus excretors showed enteroviral neutralizing antibody increments during convalescence. The dominant enteroviral serotype was coxsackievirus A9.     

Enteroviral and Mumps Meningitis in Toronto, 1963

Virological investigations of 115 children with the aseptic meningitis syndrome during 1963 resulted in the isolation of enteroviruses from cerebrospinal fluid (CSF) and/or feces of 21 of 48 children who had no association with mumps. For the third successive year, Echo 9 was the dominant enterovirus in cases of aseptic meningitis in Toronto children, but no rashes were associated with Echo 9 meningitis during 1963, in contradistinction to previous years. Mumps virus was isolated from CSF of 25 patients by inoculation of rhesus monkey kidney cultures, and rising or elevated mumps antihemagglutinin titres in paired sera from a further 33 cases provided laboratory evidence of infection with mumps virus in 58 of 67 patients with mumps meningoencephalitis. No enlargement of salivary glands was noted in 20 laboratory-proved cases of mumps meningoencephalitis. Enteroviral meningitis occurred principally during summer, but the peak of mumps meningoencephalitis occurred during late winter.     

1996年延边地区无菌性脑膜脑炎病因病毒的分离及初步鉴定

吉林省延边地区１９９６年６月发生无菌性脑膜脑炎流行,在全地区２１６万人口中,发病人数约５￣６千人,死亡２人,为历史上所罕见。从病人脑脊液和粪便标本中分离到多株病毒,分离率较高（分别达到５２．４％￣６６．７％）。用ＷＨＯ提供的ＲＩＶＭ肠道病毒组合血清进行中和定型,不能确定型别。ＲＴ－ＰＣＲ结果表明为肠道病毒。病人早期血清特异性ＩｇＭ抗性阳性率７２．０％,证明所分离的病毒为此次无菌性脑膜脑炎暴发流行的     

Aseptic meningitis,Montreal, 1969: A Clinical and Laboratory Study

During the summer and fall of 1969 an outbreak of aseptic meningitis occurred in Montreal and its environs. Forty-four patients were admitted to the infectious disease ward of The Montreal Children''s Hospital in August, September and October. Half of the patients were in the age group 6 to 10 years. The ratio of males to females was two to one. Patients showed the typical signs and symptoms of aseptic meningitis, namely fever, vomiting, headache, neck stiffness and pleocytosis of the cerebrospinal fluid.Viruses were recovered from 19 (43%) of the 44 cases. The predominant virus in the outbreak was a non-hemagglutinating strain of echovirus 6. Other virus types isolated in the outbreak were echovirus 7, coxsackievirus A9 and coxsackievirus B4. Serological investigations confirmed the validity of the echovirus isolations and identified additional cases.     

Primary Culture of Human Embryonic Kidney Cells as the Medium for Isolation of ECHO 6 Virus

A large epidemic of aseptic meningitis due to ECHO 6 virus swept over Japan in the summer months of 1965. In our studies on 89 cases of aseptic meningitis in the epidemic, primary culture of human embryonic kidney cells was shown to provide a highly sensitive host system for isolation of ECHO 6 virus from clinical materials. Virus was recovered from cerebrospinal fluid in 72% of 89 cases, from throat swabs in 64% of 36 cases, and from rectal swabs in 59% of 37 cases. Most significant is the finding that the rate of virus isolation from cerebrospinal fluid in this host system was considerably higher as compared with that obtained by other investigators in other host systems such as primary monkey kidney cells or human amniotic cells (primary or EL). This finding should be emphasized particularly because isolation of a virus from cerebrospinal fluid, in contrast to throat secretions or feces, is of much greater importance in establishing the etiologic relationship to the disease in the diagnosis of aseptic meningitis. The neutralization test was shown to be efficient in detecting ECHO 6 virus infection; acute serums should be taken preferably by the 4th day of illness and convalescent serums in the second week. Epidemiologic findings, such as the predominance of male patients and occurrence of the epidemic in summer months, generally coincide with the previous reports. However, our cases were in much younger age groups in contrast with the previous reports; 90% of our 89 patients were 6 years of age or younger, and 15 children or 17% were less than 1 year of age, including 3 less than 6 months of age. The clinical observations on our cases confirmed the previous reports.     

Interferon in the Cerebrospinal Fluid of Children with Central Nervous System Disorders

Interferon was detected in the cerebrospinal fluid (CSF) of 26 of 51 children with aseptic meningitis, two of 44 with bacterial meningitis, and four of 118 with miscellaneous conditions including encephalitis, convulsive disorders and leukemia with neurological involvement. The geometric mean titre of interferon in mumps meningitis was seven to eight times higher than that in enteroviral meningitis; however, levels of interferon were not related to the concentration of leukocytes in CSF from these patients. Interferon titres were relatively greater at the height of the febrile response in children with mumps meningitis or enteroviral meningitis. There was no association between the presence of interferon in the CSF and the isolation of mumps virus or an enterovirus from the same specimen. Patients frequently developed homologous antibody one to three days after signs of aseptic meningitis, obscuring the relationship of interferon production to clinical improvement.     

Characterization of a virus associated with epidemic conjunctivitis in Thailand.

A strain of virus, named the A-E strain, was isolated from a patient suffering from acute haemorrhagic conjunctivitis during an epidemic of the disease in Bangkok in 1972. The virus had the characteristics of an enterovirus but was not neutralized by any known enterovirus antiserum. Cross neutralization tests indicated that the isolate was closely related to the J670-71 virus isolated in Japan. The virus produced conjunctivitis in rabbits and monkeys following conjunctival inoculation.     

An Outbreak of Aseptic Meningitis due to Echo 25 Virus

In a widespread outbreak of aseptic meningitis during the summer and early fall of 1961 in Ontario, Echo type 25 virus was found to be the predominant virus type isolated from affected patients. Recovery of Echo 25 virus in several cases from the CSF of patients strongly suggests an etiological relationship between this virus type and the illness. The causative role of the virus was confirmed by demonstration of significant immunological response in patients found to be infected with the virus. The presence of antibodies to Echo 25 virus in about 70% of meningitis patients, from whom no virus was isolated, indicates that the infection in the population of the affected areas was much more prevalent than the number of successful virus isolations would suggest. It is recommended that Echo 25 virus should be added to the series of Echo virus types capable of causing outbreaks of aseptic meningitis.     

2002～2004年浙江省无菌性脑膜炎暴发疫情的病原学研究

2002～2004年浙江省发生了无菌性脑膜炎暴发疫情,为了及时查明病因,分析病原的分子特征并进行病原溯源,我们采集患者脑脊液和粪便样本271份,用RD和Hep-2细胞同时分离病毒,对分离株VP1和VP4/VP2基因测序,进行同源性与进化分析。结果从271份样本中分离到埃柯病毒30型(E30)78株;对31株分离株VP1区核苷酸(nt)序列测定,其长度均为876nt,推导编码292个氨基酸(aa)。浙江E30株与原型株Bastianni在VP1区的nt和aa同源性分别为84.7%～86.3%和92.1%～94.2%;浙江E30株之间nt和aa的同源性分别为87.1%～99.4%和96.2%～100%。在VP1基因进化树上浙江E30株分别位于G和H基因亚型分支上,与浙江E30G亚型株亲缘关系最近的国内外毒株分别为2003年江苏、山东株和1999年乌克兰株;与浙江E30H亚型株亲缘关系最近的毒株为2008年韩国株。VP4/VP2区同源性与进化分析结果与VP1相似。结果表明2002～2004年浙江省无菌性脑膜炎暴发疫情由E30G和H二类不同基因亚型流行株引起;H基因亚型株推测为新的E30变异株,首先分离于2002年浙江省。     

Oxidative Stress in Cerebrospinal Fluid of Patients with Aseptic and Bacterial Meningitis

This study aimed to determine whether patients with aseptic and bacterial meningitis presented alterations in oxidative stress parameters of cerebrospinal fluid (CSF). A total of 30 patients were used in the research. The CSF oxidative stress status has been evaluated through many parameters, such as lipid peroxidation through thiobarbituric acid reactive substances (TBARS) and antioxidant defense systems such as superoxide dismutase (SOD), glutathione S-transferase (GST), reduced glutathione (GSH) and ascorbic acid. TBARS levels, SOD and GST activity increase in aseptic meningitis and in bacterial meningitis. The ascorbic acid concentration increased significantly in patients with both meningitis types. The reduced glutathione levels were reduced in CSF of patients with aseptic and bacterial meningitis. In present study we may conclude that oxidative stress contributes at least in part to the severe neurological dysfunction found in meningitis.     

Epidemiological studies on echovirus type 18 infection in Toyama Prefecture

Echovirus type 18 (echo 18) was isolated from six aseptic meningitis children in Fukumitsu-machi, Toyama Prefecture, from July to August, 1980. This was the first virologically-confirmed epidemic of aseptic meningitis due to echo 18 in Japan. Epidemiological studies on the prevalence of this virus among the inhabitants in Toyama Prefecture were also performed. The results are summarized as follows. (1) Significant increases in neutralizing antibody titers against echo 18 were observed in all the paired sera of aseptic meningitis patients from whom echo 18 was isolated. (2) In October, 1980, echo 18 was also isolated from healthy children or from infants suffering from gastroenteritis in other areas of Toyama Prefecture. (3) Among the sera collected from 50 children (aged smaller than or equal to 12) in Fukumitsumachi in December, 1980, neutralizing antibodies against echo 18 were detected only in the younger groups (aged smaller than or equal to 8), 58.9% of these age groups showing a titer of 4 or higher. (4) Another epidemic around 1963 by echo 18 in Toyama Prefecture was retrospectively suggested by the examinations of sera collected in 1978 and 1981 from inhabitants in various areas of Toyama Prefecture. (5) Neutralizing antibody titers against the strain isolated, No. 35'80, were significantly higher than those against strain 'Metcalf,' the prototype of echo 18, in most sera including both aseptic meningitis patients and healthy inhabitants.     

Specific and nonspecific T-cell recruitment in viral meningitis: Possible implications for autoimmunity

Specific and nonspecific T-cell invasion into cerebrospinal fluid has been investigated in the nonfatal viral meningoencephalitis induced by intracerebral inoculation of mice with vaccinia virus. At the peak of the inflammatory process on Day 7 approximately 5 to 10% of the Lyt 2⁺ T cells present are apparently specific for vaccinia virus. Concurrently, in mice primed previously with influenza virus, 0.5 to 1.0% of the appropriate T-cell set located in cerebrospinal fluid is reactive to influenza-infected target cells. This vaccinia virus-induced inflammatory exudate may thus contain as many as 500 influenza-immune memory T cells. These findings are discussed from the aspect that such nonspecific T-cell invasion into the central nervous system during aseptic viral meningitis could result in exposure of potentially brain-reactive T cells to central nervous system components.     

Enteroviral aseptic meningitis in Japan, 1981-1991. A report of the National Epidemiological Surveillance of Infectious Agents in Japan.

In Japan, aseptic meningitis cases due to enterovirus infections increase every summer in various degrees with an incidence peak usually in July. During the past 11 years from 1981 through 1991, a total of 8,595 enterovirus isolations from aseptic meningitis cases were reported from 54 participating laboratories. Eight enterovirus types caused large epidemics; more than 100 isolations of each type from aseptic meningitis cases were reported for every epidemic year of the respective type. They were coxsackievirus (C) types B3 and B5, echovirus (E) types 4, 6, 7, 9, 18 and 30. Among these, the highest meningitis-associating frequency was reported for E30, representing 82.6% of the total isolations reported for the type during this period, followed by E4, 71.1%. The frequencies of E9, E7, E6 and CB5 were in a range from 54.5% to 44.4%, while that of E18 was 37.7% and that of CB3 21.0%. During the epidemics, enterovirus-associated meningitis was most frequently reported among children of 4-7 years of age. High frequencies were also shown in infants less than 1-year of age in some types. A total of 4,240 enteroviruses were isolated from cerebrospinal fluid of aseptic meningitis cases, representing 49.3% of the cases with enterovirus isolation.     

Oxidative stress in childhood meningitis: measurement of 8-hydroxy-2'-deoxyguanosine concentration in cerebrospinal fluid

To examine the involvement of reactive oxygen species, we measured the concentration of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative stress, in cerebrospinal fluid samples from 63 children with and without meningitis. We observed that the mean concentration of 8-OHdG in samples obtained during the early phase of bacterial meningitis, but not aseptic meningitis, was significantly higher than that in control samples. Clinical and laboratory improvement was associated with a fall in the 8-OHdG concentration in the patients with bacterial meningitis. Our findings suggest the presence of enhanced oxidative stress in the central nervous system of children with bacterial meningitis.     

Isolation of an Hswl Nav4 Influenza Virus from a Tufted Duck (Aythya fuligula) in Japan

A hemagglutinating agent was isolated from a tufted duck captured in Lake Shinji, Shimane Prefecture, Japan, and identified as influenza A by double immunodiffusion tests with the antiserum to influenza A virus ribonucleoprotein. The hemagglutinin of the isolate was antigenically related to that of A/New Jersey/8/76 but was not identical with it. The neuraminidase antigen of this Hswl subtype was closely related to that of A/turkey/Ontario/6118/68 and was shown to be Nav4 subtype. After experimental infection of 5-week-old SPF-chickens with the isolate, virus was recovered from various organs including the brain, despite the absence of signs of disease.     

Neurotensin-like immunoreactivity (NTLI) concentration in the cerebrospinal fluid of children and its alteration in a febrile aseptic meningitis

Neurotensin-like immunoreactivity (NTLI) concentrations in the cerebrospinal fluid (CSF) of normal children and patients with febrile aseptic meningitis, aged 7 months to 15 years, were studied. The NTLI concentrations in CSF of 27 children with normal CSF findings were 160.1 +/- 54.6 pg/ml (mean +/- S.D.). The NTLI concentration in CSF of 26 patients in an acute phase of aseptic meningitis was 110.6 +/- 51.1 pg/ml which was significantly (P less than 0.01) lower than the controls. These patients had a mean temperature of 101.4 +/- 1.5 degrees F which remained elevated for an average of 3.5 days. The NTLI concentrations in CSF of 23 patients in a recovery phase (after blood and CSF findings became normal with no fever) were 166.5 +/- 57.8 pg/ml, which did not differ significantly from the normal. There were no statistical correlations between the NTLI concentration in CSF and the protein concentration or total cell count in CSF. These results suggest that NTLI concentration changes during a febrile aseptic meningitis and that it may be associated with thermoregulation.