Experimental infection of chimpanzees with the Norwalk agent of epidemic viral gastroenteritis.

A fecal filtrate of human origin containing the Norwalk agent of epidemic viral gastroenteritis was administered by stomach tube to chimpanzees in an attempt to induce diarrheal disease. Significant postchallenge serum antibody rises against Norwalk viral antigens were demonstrated in all animals using the techniques of immune electron microscopy and radioimmunoassay. In addition, viral antigens were detected in feces from five of nine animals using radioimmunoassay. Clinical illness characterized by diarrhea and/or vomiting did not occur. Infection was transmitted subsequently by feeding four additional chimpanzees a fecal filtrate prepared from one of the previously infected animals. Development of an antibody response in four animals and detection of viral antigen in two animals that received this passage filtrate indicated that viral replication had occurred in the absence of clinical illness. The availability of the chimpanzee as an experimental animal host susceptible to infection with the Norwalk agent should facilitate the study of epidemic viral gastroenteritis.     

Outbreaks of food-borne and waterborne viral gastroenteritis.

Norwalk virus infection is the epidemiologic prototype for outbreaks of food-borne and waterborne gastroenteritis. Around the world, Norwalk virus and Norwalk-like viruses appear to be major causes of food-borne and waterborne illness. Assessment of the overall significance of viral agents to the epidemiology of food-borne and waterborne illness is hampered by the lack of surveillance throughout much of the world. In areas where food-borne and waterborne illness surveillance is conducted, outbreaks of viral gastroenteritis are underreported because of the lack of availability of routine laboratory services to confirm the viral etiology. Routine use of epidemiologic criteria as an alternative to laboratory confirmation will allow better assessments of the importance of viral gastroenteritis until effective laboratory methods can be widely implemented. Outbreaks of viral gastroenteritis have been propagated by contamination of water supplies, raw foods, and ill food handlers. Controlling an outbreak depends on identifying and removing the source of contamination. The demonstrated occurrence of person-to-person transmission and the likely occurrence of transmission of Norwalk-like viruses by aerosol make it necessary to evaluate the potential for transmission by food handlers and servers in every outbreak, regardless of primary source.     

Molecular detection and sequencing of "Norwalk-like viruses" in outbreaks and sporadic cases of gastroenteritis in Ireland

Norwalk-like viruses (NLVs) are now established as the most important causative agents of epidemic gastroenteritis worldwide. The overall objective of this study was to determine the molecular epidemiology of Irish NLV isolates for the first time by obtaining sequence data from specimens originating from outbreaks and sporadic cases of gastroenteritis. Eight samples from sporadic cases of gastroenteritis and nine isolates from separate NLV outbreaks were examined. Of the sporadic isolates, six were shown to be genogroup 2 (G2) by RT-PCR, while two were G1. All of the outbreak isolates were G2. All isolates were partially sequenced within a highly conserved region of ORF1 (RNA-dependent RNA polymerase gene). Sequence data were aligned and a dendogram was constructed. The results indicated that the majority of G2 isolates were seen to cluster with Bristol and Lordsdale virus, while the two G1 specimens were related most closely to Southampton virus. Further downstream sequence analysis of a number of the isolates confirmed this result. It is concluded that the majority of NLV isolates circulating in Ireland belong to the Bristol/Lordsdale clade.     

Induction of diarrhea in colostrum-deprived newborn rhesus monkeys with the human reovirus-like agent of infantile gastroenteritis

Summary Diarrhea developed in five newborn rhesus monkeys(Macaca mulatta) inoculated orally on the first day of life with the human reovirus-like agent of infantile gastroenteritis. Incubation period ranged from 2–5 days; virus particles were detected in stools in association with illness, and virus shedding lasted between 1 and 3 days. Virus derived from monkeys that developed illness following inoculation was infectious for other monkeys but did not induce diarrhea which could be associated temporally with virus shedding. Viral antigens were also detected in tissues of the grossly abnormal small intestine of an acutely-ill monkey. Serum antibody responses were demonstrated in two of the ill animals by complement-fixation and/or immunofluorescence.With 3 Figures     

Prevalence of Norwalk-like virus infections in cases of viral gastroenteritis among children in Osaka City,Japan

Surveillance of Norwalk-like virus (NLV) infections in cases of pediatric gastroenteritis between April 1996 and March 2000 showed that NLVs were an important causative agent in viral gastroenteritis cases among children between November and January in those years. The predominant type of NLV was closely related to Lordsdale virus in genogroup 2. During the 1999-2000 season, Arg320-like strains, which may be genetic recombinants, suddenly appeared and spread.     

一起诺如病毒性胃肠炎暴发的队列研究

目的调查分析某村一起诺如病毒性胃肠炎暴发的发病原因。方法采用历史性队列研究方法，应用ELISA对标本进行实验室检测。结果该起暴发共报告病例34例，发病与接触腹泻病人（X^2=6．51，P=0．00）、食物暴露于苍蝇（X^2=4．61，P=0．00）及村民暴露于患者呕吐物或粪便1m内（X^2=3．09，P=0．00）有统计学意义上的关联，与饮用水来源（X^2=1．68，P=0．14）没有统计学意义上的关联。在采集的8份肛拭子样本中1份ELISA法检出诺如病毒抗原阳性：4份粪便样本中2份ELISA法检出诺如病毒抗原阳性。结论这是一起由诺如病毒感染所致的急性胃肠炎暴发．密切接触是此次暴发的主要传播途径。     

Immunoglobulin M Antibody Test To Detect Genogroup II Norwalk-Like Virus Infection

Sera obtained from adult volunteers inoculated with genogroup II Norwalk-like viruses (NLVs), Hawaii virus, and Snow Mountain virus and from patients involved in outbreaks of gastroenteritis were tested for genogroup II NLV Mexico virus-specific immunoglobulin M (IgM) by use of a monoclonal antibody, recombinant Mexico virus antigen (rMXV)-based IgM capture enzyme-linked immunosorbent assay (ELISA). Sera from genogroup I Norwalk virus (NV)-inoculated volunteers and from patients involved in a genogroup I NLV outbreak were also tested. In sera from those infected with genogroup I NV or NLVs in volunteer and outbreak studies, only 3 of 25 were rMXV IgM positive; in contrast, 24 of 25 were IgM positive for recombinant NV (rNV). In sera from those infected with genogroup II NLVs in volunteer and outbreak studies, 28 of 47 were rMXV IgM positive and none were IgM positive for rNV, showing the specificity of each IgM test for its respective genogroup. In an outbreak of gastroenteritis not characterized as being of viral etiology but suspected to be due to NV, 7 of 13 persons had IgM responses to rMXV, whereas none had IgM responses to rNV, thus establishing the diagnosis as genogroup II NLV infection. The rMXV-based IgM capture ELISA developed is specific for the diagnosis of genogroup II NLV infections.     

Expression and self-assembly of recombinant capsid protein from the antigenically distinct Hawaii human calicivirus.

The Norwalk and Hawaii viruses are antigenically distinct members of the family Caliciviridae and are considered to be important etiologic agents of epidemic gastroenteritis, with most studies focusing on the role of Norwalk virus. To further investigate the importance of Hawaii virus, Hawaii virus-like particles (VLPs) were produced by expression of its capsid protein in the baculovirus system and these VLPs were used as the antigen in an enzyme-linked immunosorbent assay that was efficient in the detection of a serologic response to Hawaii virus. The ready availability of Hawaii VLPs should enable larger-scale epidemiological studies to further elucidate the importance of this agent.     

Experimental norovirus infections in non-human primates

Noroviruses, with Norwalk virus as the prototype strain, are the most common cause of viral gastroenteritis in people of all ages. Limited information on the immunology of Norovirus infections has been obtained by studies both in the natural setting and in experimentally infected volunteers. Interpretation of these studies is difficult due to the lack of information on the history of Norovirus exposure and the cross-reactivity of antibodies. An animal model for Norovirus infections would be important to study the immune response, e.g., for vaccine assessment. In the present study the susceptibility of common marmosets, cotton top tamarins, cynomolgus, and rhesus macaques to Norovirus infection was tested. Following oral inoculation, low level replication may have occurred in common marmosets and cotton top tamarins but not in cynomolgus macaques, based on short-term viral shedding; neither clinical symptoms nor antibody responses were observed in these species. In contrast, rhesus macaques were found susceptible to Norwalk virus infection as one animal shed virus for a longer period of time and developed Norwalk virus specific IgM and IgG responses. Further research on Norovirus susceptibility in rhesus macaques may yield an animal model to study the immune response and pathogenesis after Norovirus infection.     

Widespread outbreaks of clam- and oyster-associated gastroenteritis. Role of Norwalk virus

Consumption of raw shellfish has long been known to be associated with individual cases and sporadic outbreaks of enteric illness. However, during 1982, outbreaks of gastroenteritis associated with eating raw shellfish reached epidemic proportions in New York State. Between May 1 and December 31, there were 103 well-documented outbreaks in which 1017 persons became ill: 813 cases were related to eating clams, and 204 to eating oysters. The most common symptoms were diarrhea, nausea, abdominal cramps, and vomiting. Incubation periods were generally 24 to 48 hours long, and the duration of illness was 24 to 48 hours. Bacteriologic analyses of stool and shellfish specimens did not reveal a causative agent. Norwalk virus was implicated as the predominant etiologic agent by clinical features of the illness and by seroconversion and the formation of IgM antibody to Norwalk virus in paired serum samples from persons in five (71 percent) of seven outbreaks in which testing was done. In addition, Norwalk virus was identified by radioimmunoassay in clam and oyster specimens from two of the outbreaks. Determining the source of the shellfish was not always possible, but northeastern coastal waters were implicated. The magnitude, persistence, and widespread nature of these outbreaks raise further questions about the safety of consuming raw shellfish.     

Localization of simian immunodeficiency virus in the central nervous system of rhesus monkeys.

Simian immunodeficiency virus (SIV), like the human immunodeficiency virus (HIV), is a lentivirus that is both immunosuppressive and neurovirulent. Rhesus macaques (Macaca mulatta) inoculated with SIV often develop a giant cell encephalitis similar to that seen in humans infected with HIV. The authors examined SIV expression by immunohistochemistry and RNA in situ hybridization in the cerebrum, cerebellum, choroid plexus, and spinal cord from five macaques with and two macaques without giant cell encephalitis. Selected portions of the central nervous system (CNS) also were examined by electron microscopy. Simian immunodeficiency virus was detected in the CNS of all seven monkeys whether or not they had giant cell encephalitis. Both SIV antigen and RNA were present in all levels of the CNS examined. Macrophage/giant cell lesions always contained viral RNA and antigen and were the only sites where viral particles were detected by electron microscopy. However, SIV antigen and RNA also were commonly associated with small vessels, the choroid plexus, and meninges; these were the only locations where virus was detected in animals without giant cell encephalitis. Immunophenotyping showed that the cellular infiltrates consisted primarily of monocyte/macrophages and occasional CD8-positive T cells. Macrophages and T cells also were present in the stroma of the choroid plexus and were intimately associated with vessels in the CNS of SIV-infected but not uninfected macaques. Simian immunodeficiency virus infection of the macaque CNS provides an excellent model for studying the pathogenesis, treatment, and prevention of HIV-1-encephalitis.     

Antigenic diversity of Norwalk-like viruses: expression of the capsid protein of a genogroup I virus, distantly related to Norwalk virus

Summary.  The gene encoding the capsid protein of a genogroup I Norwalk-like virus (NLV) (Hu/NLV/Stav/95/Nor) was cloned and expressed in insect cells using a baculovirus vector. The His-tagged recombinant capsid protein (rStav) was antigenic and immunogenic, showed an apparent molecular weight of approximately 68 kD in protein gels, and was only soluble under denaturing conditions. The amino acid sequence of the rStav protein showed 65–88% similarity to capsid protein sequences from other genogroup I NLV and was most closely related to Desert Shield virus. Norwegian recruit sera were tested for antibodies against rStav by Western blotting (rStav WB). The sera had previously been tested for antibodies against a recombinant Norwalk virus capsid protein in an ELISA (rNV ELISA). Several rNV ELISA-negative sera showed a positive response in the rStav WB, indicating that the use of antigens representing different stains may be necessary when screening sera for antibodies against genogroup I NLV. Received July 7, 1999/Accepted November 9, 1999