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The experimental production of diarrhoea in colostrum deprived axenic and gnotoxenic calves with enteropathogenic Escherichia coli, rotavirus, coronavirus and in a combined infection of rotavirus and E. coli.

We attempted to produce diarrhoea experimentally in the newborn calf by orally injecting 17 colostrum-deprived calves with two serotypes of Escherichia coli Ent+ K99+, a rotavirus and a coronavirus. With E. coli alone, a dose of 2 x 10(8) bacteria administered 24 hours after birth causes a mild attack of diarrhoea, whereas 1 x 10(10) bacteria leads to dehydration and death. An inoculation of rotavirus is followed by diarrhoea which always contains large quantities of rotavirus. These animals were anorectic for a time, but none was dehydrated or died. With coronavirus, there were large quantities of watery diarrhoea, which led to dehydration and death. The inoculation of rotavirus, not lethal in itself, followed by a similarly non lethal inoculation of E. coli in doses of 3 x 10(8) to 2 x 10(9) led to dehydration and death. The authors conclude that dehydration and death of the animal can caused by large doses of E. coli or coronavirus or by two non-lethal doses of rotavirus and E. coli administered one after the other.

Animals↗

Human rotavirus infection in Malaysia. III. A one year survey on the prevalence of rotavirus enteritis in children.

The prevalence of human rotavirus enteritis in children admitted to the gastroenteritis ward of the Kuala Lumpur General Hospital was studied in 1982. Human rotavirus in the stool of the patients was detected by enzyme-linked immunosorbent assay. The survey showed that rotavirus enteritis in children were observed throughout the year, with two broad peaks of rotavirus infection occurring around March and September. The lowest incidence was recorded in July, however, no prolonged period of low prevalence of rotavirus enteritis was observed. The average prevalence for the whole of 1982 was 40% of the total diarrhoeal cases. No significant relationship was noted between the prevalence of the disease and rainfall.

Child, Preschool↗

Rotavirus and rotavirus vaccines.

Two new rotavirus vaccines have recently been licensed that will provide the intervention needed to diminish the huge burden of rotavirus disease among all children. In many upper and middle income countries, these vaccines will soon be available for the routine immunization of children. The impact should be a rapid and measurable reduction in hospitalizations and doctor visits for acute diarrhea in children, especially in the winter rotavirus season. While few deaths occur in these settings, the illness has consequences in terms of both medical costs and indirect costs, including parents work time lost. In the developing world, clinical trials are still needed to ensure that the vaccines being licensed will work as expected in children living in poor settings. In these settings, other enteric flora, micronutrient malnutrition, higher titers of maternal antibody and other factors still poorly defined have compromised other live oral vaccines and have required the developers to alter vaccine formulation, dose, or schedule. Until these trials are completed, we can only hope that the efficacy will be comparable and that the vaccine will prove to be life-savers. Once the efficacy and safety have been established, rotavirus vaccines could provide a major boost to programs for child survival.

Animals↗

Economic evaluation of rotavirus vaccinations in Finland: randomized, double-blind, placebo-controlled trial of tetravalent rhesus rotavirus vaccine.

The cost-benefit ratio of tetravalent rhesus rotavirus vaccine (RRV-TV) in Finland for prevention of rotavirus gastroenteritis was assessed in a randomized, double-blind, placebo-controlled trial. Costs related to vaccination, side effects, and gastroenteritis were identified. Children received RRV-TV (n = 1,191) or placebo (n = 1,207) at 2, 3, and 5 months of age with other infant vaccinations. Prospective follow-up averaged 1.0 years per child. An intention-to-treat analysis was performed from the perspective of society. Nine cases of severe rotavirus gastroenteritis occurred in the RRV-TV group, versus 100 in the placebo group (P < .0001); mean cost per vaccinated child was 4 Finnish marks (FIM) in the RRV-TV group, versus 203 FIM in the placebo group. Side effects with related costs occurred after 11% and 7% of doses in the RRV-TV group and placebo group, respectively (P < .001); mean cost per child was 89 FIM vs. 75 FIM. The break-even cost (i.e., net benefit, excluding cost of vaccine) of RRV-TV in prevention of severe rotavirus gastroenteritis was 109 FIM (U.S. $19.60) per child.

Cost-Benefit Analysis↗

Prevention of rotavirus disease: guidelines for use of rotavirus vaccine.

On February 3, 2006, a bovine-based pentavalent rotavirus vaccine (RotaTeq, Merck & Co Inc, Whitehouse Station, NJ) was licensed by the US Food and Drug Administration for use in infants in the United States. The American Academy of Pediatrics recommends routine immunization of infants with 3 doses of pentavalent rotavirus vaccine administered orally at 2, 4, and 6 months of age. The first dose should be administered between 6 and 12 weeks of age; immunization should not be initiated for infants older than 12 weeks of age. Subsequent doses should be administered at 4- to 10-week intervals, and all 3 doses of vaccine should be administered by 32 weeks of age. Pentavalent rotavirus vaccine can be coadministered with other childhood vaccines. Pentavalent rotavirus vaccine is contraindicated for infants with a serious allergic reaction to any vaccine component or to a previous dose of vaccine.

Administration, Oral↗

Detection of anti-rotavirus IgG, IgM, and IgA antibodies in healthy subjects, rotavirus infections, and immunodeficiencies by immunoblotting.

Immune responses to individual simian rotavirus (SA 11) structural polypeptides were studied with emphasis on specific IgG, IgM, and IgA in paired sera from four children with rotavirus infections. Responses to simian rotavirus (SA 11) were also studied in 103 healthy children, 10 patients with primary immunodeficiency who received intravenous immunoglobulin, and 11 human immunodeficiency virus antibody-positive patients. All samples were immunoblotted for two major polypeptides--VP2 and VP6--of IgG. Immunoblotted IgA and IgM were elevated 2-4 weeks after the onset of the disease (stage II) in patients with rotavirus infection. Immunoblotted IgG remained almost the same at the onset of the disease and stage II. The scores for primary and secondary immunodeficiency patients were lower than for healthy children. The score obtained by totaling all polypeptides found in each individual was compared with the optical density (OD) value obtained by enzyme immunoassay.

Acquired Immunodeficiency Syndrome↗

Comparison of reactogenicity and antigenicity of M37 rotavirus vaccine and rhesus-rotavirus-based quadrivalent vaccine.

90 Venezuelan infants aged 10-20 weeks were randomly allocated to four groups which received one of the following: the M37 vaccine (1 x 10(4) pfu [plaque-forming units]); quadrivalent rotavirus vaccine (1 x 10(4) pfu each of serotype 3 rhesus rotavirus [RRV] and human rotavirus-RRV reassortants of serotypes 1, 2, and 4); balanced quadrivalent vaccine consisting of 1 x 10(4) pfu of serotype 1 and 3 components but 5 x 10(4) pfu of serotype 2 and 4 components; or placebo. The frequencies of transient febrile responses in these four groups were 20%, 27%, 30%, and 9%. 50% of 22 infants tested who received M37 vaccine showed a serum rotavirus IgA antibody response, compared with 74% of the 23 quadrivalent and 86% of the 22 balanced-quadrivalent recipients. 64% of the M37 recipients showed a neutralising antibody response to M37; 27% showed such responses to human serotype 1 Wa strain and 27% to serotype 4 neonatal strain ST3. 17-39% of the quadrivalent recipients and 27-41% of the balanced-quadrivalent recipients showed neutralising antibody responses to serotypes 1-4. 70-73% of the quadrivalent and balanced quadrivalent groups also showed neutralising antibody responses to RRV.

Antibodies, Viral↗

Vaccinia-rotavirus VP7 recombinants protect mice against rotavirus-induced diarrhoea.

Recombinant vaccinia viruses expressing wild type intracellular VP7 (VP7wt) from rotavirus SA11 or VP7sc, a cell surface-anchored variant, boosted antibody titres in SA11-immune mice. Pups born to these mice were protected from diarrhoea following challenge with SA11. In rotavirus-naive mice, two immunizations with recombinant vaccinia virus expressing VP7sc stimulated protective immunity that could be transferred to pups, whereas viruses expressing VP7wt did not stimulate protective immunity. Recombinant vaccinia viruses expressing intracellular or cell surface-anchored VP6, the rotavirus group-reactive antigen from the inner capsid, did not stimulate protective immunity. These experiments demonstrate that a live viral vector expressing cell surface anchored VP7 may represent a strategy for the development of safe, effective vaccines against rotavirus-induced diarrhoea.

Animals↗

Novel porcine rotavirus of genotype P[27] shares new phylogenetic lineage with G2 porcine rotavirus strain.

A novel and unusual strain of porcine rotavirus (PoRV) CMP034 was isolated from a 7-week-old piglet during the epidemiological survey of porcine rotavirus infection in Chiang Mai province, Thailand from June 2000 to July 2001. Molecular characterization of gene VP4 by sequence analysis showed a low level of amino acid sequence identity, ranging from 56.7% to 76.6%, while comparison of VP8* portion showed 41.8% to 69.9% identity, with the 26 P genotypes recognized to date. Phylogenetic analysis of the VP4 sequence revealed that CMP034 was only distantly related to the other 26 P genotypes and was located in a separate branch. Sequence analysis of gene VP7 showed the highest level of amino acid identity (94.7%) with the PoRV G2-like reference strain 34461-4 but a lower level of identity with those of human G2 rotaviruses, ranging from 87.7% to 88.0%. Phylogenetic analysis of gene VP7 revealed two major lineages among G2 rotavirus strains based on the host origin. PoRV strain CMP034 clustered exclusively with G2-like PoRV strain 34461-4 in a novel lineage that is distinct from the major G2 human lineage. Moreover, strain CMP034 displayed a porcine-like VP6 and NSP5/6 with subgroup I specificity, while bearing an NSP4 with some genetic group B human-like characteristics. These findings provide evidence that CMP034 should be considered as a novel VP4 genotype P[27].

Amino Acid Sequence↗

Detection of rotaviruses by nucleic acid hybridization with cloned DNA of simian rotavirus SA11 genes.

We developed a dot-blot hybridization assay to detect rotaviral RNA sequences in tissue culture or in clinical samples. 32P-labeled cloned cDNA probes of the simian rotavirus SA11 specifically detected rotaviral RNA sequences and were more sensitive for detecting SA11 than was the commercial enzyme-linked immunosorbent assay Rotazyme test. A full-length probe of SA11 gene 6 detected 2.5 X 10(5) SA11 particles or approximately 0.27 ng of purified SA11 dsRNA. Combined probes from genes 6 and 9 detected 0.135 ng of purified SA11 dsRNA. The assay detected group A rotaviruses from different subgroups and serotypes, but the sensitivity of RNA detection varied from 0.5 to 31 ng when RNA from heterologous strains of virus was analyzed. An analysis of coded stool samples correctly identified 31 (91%) of 34 samples positive for rotavirus by electron microscopy and 100% of 26 samples negative for rotavirus by electron microscopy. Preliminary experiments also showed the assay has potential to directly characterize (subgroup and serotype) rotaviral isolates.

Cloning, Molecular↗

Comparative evaluation of reactogenicity and immunogenicity of two dosages of oral tetravalent rhesus rotavirus vaccine. US Rhesus Rotavirus Vaccine Study Group.

OBJECTIVE: To compare the safety and immunogenicity of two dosages of tetravalent rhesus rotavirus vaccine (RRV-TV) and the effect of age at dosing. METHODS: A total of 195 infants were stratified by age into 2 groups, 6 to 12 weeks and 16 to 24 weeks, and randomly assigned to receive a single dose of placebo or RRV-TV containing either 4 x 10(5) or 4 x 10(6) plaque-forming units (pfu). Symptoms were recorded for 5 days after vaccination. Anti-rotavirus IgA and neutralizing antibody to human rotavirus serotypes G1 to G4 and RRV were measured in serum obtained pre- and postvaccination. RESULTS: Rates of fever > 38 degrees C (9%), diarrhea (6%) and vomiting (8%) were similar in all groups. IgA (69% vs. 49%, P = 0.02) and RRV (85% vs. 66%, P = 0.004) seroconversion rates were significantly higher in the 4 x 10(6) pfu vaccine group as were antibody titers to RRV (440.2 vs. 263.7, P = 0.04). Older infants demonstrated significantly higher seroconversion rates and antibody titers for IgA (71% vs. 52%, P = 0.03; and 110.6 vs. 54.8, P = 0.004) and RRV (92% vs. 66%, P = 0.05 and 498.3 vs. 205.6, P = 0.01) at either dose level than did the younger infants. There were no significant differences in seroconversion rates or antibody titers to human rotavirus types G1 to G4 between the two vaccination groups. CONCLUSIONS: RRV-TV at a dose of 4 x 10(6) pfu can be safely administered to infants 6 to 24 weeks of age. A single dose of 4 x 10(6) pfu of RRV-TV was significantly more immunogenic than a single dose of 4 x 10(5) pfu but did not improve responses to the human serotypes. Older vaccine recipients demonstrated significantly higher IgA and neutralizing antibody seroconversion rates and antibody titers than younger infants independent of dosage.

Administration, Oral↗

Shedding of rotavirus after administration of the tetravalent rhesus rotavirus vaccine.

BACKGROUND: The tetravalent rhesus rotavirus vaccine (RV-TV) has been administered to several thousand children in multiple settings throughout the world. It has been proved safe and efficacious, resulting in an application for licensure in 1997. However, only limited information has been reported on viral shedding after RV-TV vaccination. METHODS: Stool specimens were collected between Days 3 and 5 after administration of each of 3 doses of RV-TV to 248 subjects 6 to 12 weeks of age in 8 centers across the United States. Rotavirus antigen was measured by an enzyme-linked immunosorbent assay to determine the number of subjects who shed after each dose. The relative quantities of vaccine strains shed were then determined by plaque purification and serotype analysis. RESULTS: Rotavirus shedding was detected in 125 (50.4%) subjects, and 19 shed after more than 1 dose. Although fewer subjects shed rotavirus after Dose 2 (14.5%), shedding after Doses 1 (26.0%) and 3 (22.5%) were comparable. After plaque purification and serotyping, most viruses shed (76.2%) were identified as G3 after Dose 1, but a major shift to G2 strains was found after Doses 2 (61.3%) and 3 (69.0%). CONCLUSIONS: Sequential RV-TV administrations caused no overall significant decrease in the number of vaccinees who experienced detectable shedding. A major shift in shedding was found from the serotype G3 vaccine strain (RRV) after the first dose of vaccine to the serotype G2 reassortant after Doses 2 and 3.

Humans↗

Independent segregation of the VP4 and the VP7 genes in bovine rotaviruses as confirmed by VP4 sequence analysis of G8 and G10 bovine rotavirus strains.

We determined the complete nucleotide sequences of the VP4 genes of five bovine rotavirus strains (A5, 61A, A44, B223 and KK3). The deduced VP4 amino acid sequence of strain A5 with G8 serotype specificity is 91.1% identical to that of strain NCDV of the serotype G6, and strain 61A with G10 serotype specificity has a VP4 amino acid sequence (95.2% identity) similar to that of the UK strain of the G6 serotype. In contrast, the VP4 amino acid sequences of strains A44, B223 and KK3 of the G10 serotype, isolated in Thailand, the U.S.A. and Japan respectively, have very similar sequences to each other, but less similarity (50 to 60%) to other group A rotavirus strains reported so far. Their VP4 genes are 2352 nucleotides in length and encode 772 amino acids, four amino acids fewer than the VP4 proteins of other animal rotaviruses. Thus, the presence of three different VP4 types (P types) and the independent segregation of G serotype and P type were postulated in bovine rotaviruses by VP4 sequence analysis.

Amino Acid Sequence↗

Isolation, identification and characterization of group A rotavirus from a chicken: the inner capsid protein sequence shows only a distant phylogenetic relationship to most other avian group A rotaviruses.

Rotavirus particles were identified in the intestinal content of a 35-day-old stunted chicken. The virus was isolated, RNA pattern was analysed and the viral genome segment 6 was sequenced. In particular, the sequence data showed a very close similarity to the chicken rotavirus isolate Ch-1 (99.2% amino acid homology), this is distantly related to all known avian rotaviruses and supports the existence of different VP6 types amongst avian group A rotaviruses.

Animals↗

New porcine rotavirus serotype antigenically related to human rotavirus serotype 3.

Serotyping of porcine rotaviruses isolated in MA104 cells from Australian piglets with diarrhea showed that two strains belonged to serotype 3 and one strain was antigenically similar to the OSU strain of porcine rotavirus (serotype 5). In addition, neutralizing antibodies to human rotavirus serotype 4 (ST-3 strain) were detected in serum samples from sows in one area, and so it seems probable that porcine rotaviruses of at least three serotypes occur in Australia.

Animals↗

Evaluation of the ImmunoCardSTAT! rotavirus assay for detection of group A rotavirus in fecal specimens.

Rapid detection of group A rotavirus was performed by using ImmunoCardStat! Rotavirus (ICS-RV) (which uses immunogold-based, horizontal-flow membrane technology), two commercial enzyme immunoassays (Premier Rotaclone and TestPack Rotavirus), and electron microscopy. A total of 249 stool specimens collected from children with gastroenteritis between February and April 1997 were tested. After resolution of 19 of the 22 discordant results by reverse transcription-PCR for group A rotavirus, ICS-RV detected 125 positives while Rotaclone and TestPack detected 127 and 129 positives, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value were 94.0, 100, 100, and 93.4% for ICS-RV; 95.5, 100, 100, and 95.0% for Rotaclone; and 97.0, 96.5, 97.0, and 96.5% for TestPack. ICS-RV was sensitive and specific and was relatively simple to perform and interpret.

Adolescent↗

Great diversity of group A rotavirus strains and high prevalence of mixed rotavirus infections in India.

We previously observed a marked diversity of rotavirus strains and a high prevalence of the uncommon serotype G9 in a small survey of rotavirus strains collected from six centers in India. In the present study, we characterized a larger collection of strains from children hospitalized with severe diarrhea in seven Indian cities between 1996 and 1998. A total of 287 strains were G and P genotyped by reverse transcription-PCR, and some were further characterized by electropherotyping and subgrouping. Of the four strains common globally, three were found in only 43% of samples (P[8], G1, 15%; P[4], G2, 22%; P[8], G4, 6%), whereas G9 strains made up 17% of the total. Three different G9 strains were present: a P[8], G9 strain, which displayed the long electropherotype and subgroup II VP6 specificity, and two P[6], G9 strains, one with the long electropherotype and subgroup II specificity and the other with the short electropherotype and subgroup I specificity. Marked diversity was observed among strains collected from different cities and collected over time. Of the 253 strains that were fully typed, 54 (21%) had a mixed G or P genotype. Serotype G2 strains were detected more often in infections caused by single strains than in mixed infections (P < 0.05), whereas serotype G1 strains were found more often in mixed infections than in infections caused by single strains (P < 0.05). The diversity of rotavirus strains and the high prevalence of mixed infections confirm trends reported earlier and help to better characterize the strains of rotavirus circulating in India. Vaccines under development should clearly target G9 strains, and G9 should be included as one of the common global serotypes.

Child↗

Rotavirus gastroenteritis and central nervous system (CNS) infection: characterization of the VP7 and VP4 genes of rotavirus strains isolated from paired fecal and cerebrospinal fluid samples from a child with CNS disease.

Rotavirus RNA was detected in the cerebrospinal fluid (CSF) of a child with central nervous system disease symptoms associated with rotavirus gastroenteritis. The rotavirus isolates from the fecal and CSF samples were genotyped as G1P[8]. Sequence analysis of the VP7 and VP4 proteins derived from the fecal and CSF samples were remarkably similar to each other and to G1P[8] rotavirus strains commonly circulating in the community and associated with gastroenteritis.

Antigens, Viral↗