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Conclusion: Among 20 patients with inner ear complications and/or peripheral facial palsy secondary to acute otitis media (AOM) a proven or probable bacteriological cause was found in 13 (65%). In seven patients (35%), a proven or probable viral cause was found. Only two of the patients (10%), with a proven bacterial AOM and a clinical picture of a purulent labyrinthitis in both, together with a facial palsy in one, had a substantial degree of dysfunction. Although the number of patients in this study is relatively low our findings show that inner ear complications and facial palsy due to AOM can be of both bacterial and viral origin. Severe sequelae were found only where a bacterial origin was proven., Objectives: Inner ear complications and/or peripheral facial palsy secondary to AOM are rare. The general understanding is that they are due to bacterial infections. However, in some of these patients there are no clinical or laboratory signs of bacterial infections and they have negative bacterial cultures. During recent years different viruses have been isolated from the middle ear or serologically proven in AOM patients and are thought to play a pathogenetic role. We suggest that in some cases of AOM complications from the inner ear and the facial nerve can be caused by viruses. The purpose of our study was to analyze infectious agents present in patients with inner ear complications and/or facial palsy arising from AOM., Patients and Methods: The medical records of 20 patients who had inner ear complications and/or facial palsy following AOM ( unilateral in 18, bilateral in 2) between January 1989 and March 2003 were evaluated. Bacterial cultures were carried out for all patients. Sera from 12 of the patients were stored and tested for a battery of specific viral antibodies. In three patients, investigated between November 2002 and March 2003, viral cultures were also performed on samples from the middle ear and nasopharynx., Results: Nineteen patients had inner ear symptoms. Eight of them had a unilateral sensorineural hearing loss and vertigo, three had vertigo as an isolated symptom and one, with bilateral AOM, had bilateral sensorineural hearing loss. Seven patients had a combination of facial palsy and inner ear symptoms (unilateral sensorineural hearing loss in three, unilateral sensorineural hearing loss and vertigo in two, bilateral sensorineural hearing loss and vertigo in one, with bilateral AOM, and vertigo alone in one). One patient had an isolated facial palsy. Healing was complete in 11 of the 20 patients. In seven patients a minor defect remained at follow-up (a sensorineural hearing loss at higher frequencies in all). Only two patients had obvious defects (a pronounced hearing loss in combination with a moderate to severe facial palsy (House-Brackman grade 4) in one, distinct vestibular symptoms and a total caloric loss in combination with a high-frequency loss in the other. Eight patients had positive bacteriological cultures from middle ear contents: Streptococcus pneumoniae in two, beta-hemolytic Streptococcus group A in two, beta-hemolytic Streptococcus group A together with Staphylococcus aureus in one, Staph. aureus alone in one and coagulase-negative staphylococci (interpreted as pathogens) in two. In the 12 patients with negative cultures, there was a probable bacteriological cause due to the outcome in SR/CRP and leukocyte count in five. In four patients serological testing showed a concomitant viral infection that was interpreted to be the cause (varicella zoster virus in two, herpes simplex virus in one and adenovirus in one.) In three there was a probable viral cause despite negative viral antibody test due to normal outcome in SR/CRP, normal leukocyte count, serous fluid at myringotomy and a relatively short pre-complication antibiotic treatment period.

Objective: To investigate whether a solution of oxybuprocaine hydrochloride, 0.4%, results in a false-positive response in an immunochromatographic SAS Adeno Test., Design: Experimental study., Controls: Physiologic saline and 2% lidocaine., Testing: Each chemical (100 microl) was diluted in a transport medium. Five drops (200 microl) of the resultant solution were dispensed into the round sample well of a test device. Fifteen samples were tested in each group., Main Outcome Measures: Ten minutes after the start of the test, a colored line in the "specimen" portion of the test membrane was visually read as positive or negative by a masked technician., Results: No positive reaction was observed in the control groups (physiologic saline and lidocaine). A false-positive reaction was observed in six samples (33.3%) in the oxybuprocaine group. The positive rate was significantly higher in the oxybuprocaine group compared with those in control groups (P = 0.0062, Fisher's extract probability test)., Conclusions: Oxybuprocaine may induce a false-positive reaction in an immunochromatographic SAS Adeno Test. We recommend the use of lidocaine, instead of oxybuprocaine, for local anesthesia in taking eye swabs from patients with suspected adenovirus infection.

A large outbreak of adenovirus type 4-associated acute respiratory disease (ARD) occurred at Fort Jackson, South Carolina, in 1997. A laboratory-based ARD surveillance program was initiated at Fort Gordon, Georgia, where advanced individual training was heavily populated with Fort Jackson soldiers. Adenovirus type 4 was isolated from 50% of 147 trainees hospitalized with ARD. Most (88%) introduced cases were in trainees from Fort Jackson.

Haemophilus influenzae and Streptococcus pneumoniae are common causes of respiratory tract infections. H. influenzae attach to receptor epitopes in mucins and in epithelial cell membranes. Attachment is followed by an epithelial cell cytokine response. Secreted cytokines then initiate inflammation, upset the integrity of the mucosal barrier, and lead to disease. S. pneumoniae do not bind to mucins but attach to respiratory tract epithelial cells. Attachment is increased by viral infection of the epithelial cells. Unlike H. Influenzae, S. pneumoniae induce apoptosis in epithelial cells, thus disrupting the mucosal barrier. Attachment and persistence is counterbalanced by antiadhesive as well as bactericidal molecules in secretions such as human milk. These examples illustrate the balance between host defenses and microbial virulence as it has coevolved to maintain the health of the respiratory mucosa.

Purpose: The purpose of this study was to determine if the implementation of a formal set of infection-control policy and procedures (ICPPs) can reduce the number of outbreaks of epidemic keratoconjunctivitis (EKC) and the number of nosocomially infected patients in a large teaching eye institute., Methods: A retrospective and prospective study of the incidence of EKC and the number of affected patients was performed for the years 1984 through 1991. Infection-control measures (ICPPs) were formulated in 1992 with regulations implemented for patient control and management, hand washing, instrument disinfection, medication distribution, and employee furloughs. Two levels of ICPPs were established on the basis of nonepidemic or epidemic conditions. After implementation of ICPPs, a prospective 4-year study (1992 through 1995) and statistical analysis were performed to determine whether the number of outbreaks of EKC and affected patients significantly decreased., Results: The incidence of institutional EKC epidemics per year was at least one and as many as three from 1984 through 1991. After implementation of a formal set of ICPPs, no epidemics occurred in 2 of 4 years studied. The number of epidemics and affected patients was significantly less when the years before and after implementation of ICPPs were compared by chi-square analysis (P < .01 and P <. 01, respectively)., Conclusions: In this first prospective study of institutional outbreaks of EKC, the implementation of ICPPs was demonstrated to be an effective means to decrease the number of EKC outbreaks and nosocomially infected patients for this particular institution. Although several reports of institutional outbreaks of EKC have described infection-control measures that eventually controlled an outbreak well under way, this study provides policies and procedures that may effectively decrease the number and size of nosocomial epidemics of adenoviral conjunctivitis in large teaching eye institutions.

Adenovirus infection is a rare, but serious infection, during the neonatal period. The actual model of infection at birth is currently unknown, however, several mechanisms have been proposed. We describe a case of fatal neonatal adenovirus pneumonia in a 25-wk gestational age infant. Adenovirus was confirmed by electron microscopy and by in situ hybridization. The maternal cervical/endocervical smear taken prior to the delivery contained epithelial cells with changes suggestive of adenovirus, which was confirmed by in situ hybridization on the smear. These findings suggest that ascending viral infection is a factor in the pathogenesis of neonatal adenovirus infection. The identification and reporting of adenovirus may be important during pregnancy in order to avoid delay in delivery of the fetus once membranes have ruptured.

Understanding the pathogenesis of viral myocarditis is linked to the availability of sensitive assays to detect viruses in clinical material. Recent advances in molecular techniques permit direct detection of viral-specific nucleic acid in tissue samples. This report describes a protocol for DNA extraction and amplification of adenovirus genome from formalin-fixed, paraffin-embedded human tissues that detects as little as 10 copies of viral genome in a background of 0.5 micrograms of human DNA. This sensitive assay permitted the examination of archived tissues to establish a retrospective diagnosis of adenoviral myocarditis in two pediatric patients.

Antibodies to the RNA polymerase III transcription termination factor La are frequently found in the serum of patients with various autoimmune diseases. The mechanisms by which autoimmune responses are evoked remain largely obscure, but the presentation of autoantigens on the cell surface during stress conditions has been reported as a possible factor. In this study we analysed the effects of adenovirus infection on the binding of anti-La antibodies to the surface of several human cell lines and on the levels of the membrane-expressed glycoproteins HLA class I, CD44 and the CD3 complex. In addition, we studied the relative amount and the intracellular distribution of the La protein as well as its association with the major species of non-coding virus-associated (VAI) RNA. While immunofluorescence patterns revealed a redistribution and possibly cell surface expression of the La protein during infection, this could not be confirmed by other techniques. In contrast, surface levels of HLA class I proteins and CD3 complex were severely affected. The data suggest that the subcellular distribution of the La protein is not detectably influenced by adenovirus infection.

The distribution of adenovirus types in faecal samples of patients with suspected viral gastroenteritis from South Australia was determined during the 12-month period, July 1991-June 1992. There were 3299 samples tested and 226 (6.9%) were positive for adenovirus by enzyme immunoassay. Of these 226 samples, 154 (68%) were typed directly using virus DNA extracted from the faecal samples according to the Sma I, Hind III and BstE II restriction patterns and Southern hybridization analysis with pooled viral genomic DNA probes. In this group, 86% of the samples were from patients who were < 3 years of age. Enteric adenovirus types 40 and 41 accounted for 20 and 40% respectively, of these samples, and types 1, 2, 3, 5, 6, 7 and 31 comprised the remainder. Type 40 was detected mainly in the winter and spring periods, and type 41 predominated in the autumn period. The majority of the non-enteric types were found during the late winter and spring periods.

Adenovirus isolates from 52 patients with ocular infection over a 3-year period were typed by restriction endonuclease analysis in a clinical laboratory. The results indicated that adenovirus type 8 was the most common cause of adenovirus eye infection during this period, being responsible for 42 (81%) of the 52 cases. Of 42 adenovirus type 8 isolates, 22 showed variant patterns by restriction endonuclease analysis and required multiple enzyme digests for identification. These isolates were readily identified by neutralisation tests.

Forty-three strains of adenovirus type 3 isolated from patients in Norway between 1970 and 1991 were analyzed with four restriction endonucleases. Bg1 II was the most discriminative enzyme. Five genotypes were identified and one of these has not been described before (Ad3a12). During both the epidemics in this period, new genotypes were introduced into the population. The same genotypes were identified in Norway as have previously been found in the northern parts of Europe, America and the Soviet Union.

Five hundred eighty-seven adenovirus type 3 (Ad3) isolates were established from children with acute respiratory infections (ARI) from 1986 to 1991, in Yamagata, Japan. Ad3 could be found in almost all the months during the 6 years when two epidemics occurred, in 1987 and 1989. A molecular epidemiological study was done on 346 of the 587 isolates, using restriction endonucleases; BamHI, HindIII, SmaI, and BgIII were used. The Ad3 isolates were classified into seven genome types. The genetic differences among the seven genome types were < 0.9%, and their phylogenetic tree, estimated by the neighbor-joining method, correlated highly with their monthly distribution. One genome type predominated for 56 months, while the other six related genome types cocirculated for a short period. These results suggested that the predominant genome type of Ad3 might have been endemically perpetuated in the Yamagata area with minor genomic variations. Furthermore, the outbreaks of Ad3 may have been due not to the appearance of a new genome type but rather to the endemic genome type.

In a study designed to evaluate the genetic variability of adenovirus strains associated with infantile cases of respiratory disease requiring hospitalization, a collection of 136 adenovirus isolates obtained in the Roberto del Rio Children's Hospital of Santiago, Chile between June 1988 and November 1990 was studied by restriction enzyme analysis. Nasopharyngeal aspirates were obtained on admission from children under 2 years. During the study period a total of 227 adenovirus respiratory infections (ARI) were diagnosed at the ward for ARI by immunofluorescence, representing 23% of all admissions. Fifty percent of the 136 typed strains were found to belong to subgenus B, and the other 50% corresponded to subgenus C. Digestion with a set of seven enzymes allowed the identification of nine different genome types of subgenus C, three of which had not been previously described, exhibiting novel restriction patterns with either BgI II or BstEII. Ad7h, identified in 66 isolates, was the predominant genome type and was associated with the nine cases requiring mechanical respiratory assistance and with the two fatalities recorded during the 29 months. No differences were found between the age and sex distribution of subgenus B and C genomic variants, but the mean length of hospital stay (X +/- 2 SE) recorded among patients infected with subgenus B types was significantly higher (17.72 + 4.52 days (n = 55) vs. 7.54 + 1.70 days (n = 53); F = 17.22; P < 0.0001).

The adenovirus (Ad) early region 3 (E3) glycoprotein of 19K (gp19K) binds major histocompatibility (MHC) class I antigens in the endoplasmic reticulum (ER), and the gp19K-class I complex is retained in the ER through an ER retention signal at the C-terminus of gp19K. This retention of class I antigens blocks cytolysis of gp19K-expressing cells by cytotoxic T lymphocytes (CTL). Animal models infected with Ad mutants lacking gp19K support a role for gp19K in counteracting a CTL response. Gp19K binds with different avidities to different class I antigens, and portions of the gp19K sequence are highly variable among Ad serotypes in different subgroups (Ad3, 11, and Ad35 in subgroup B; Ad2 and Ad5 in subgroup C); this raises the possibility that certain human individuals may be more susceptible to productive or persistent infection by particular serotypes of Ad, depending on the haplotype of the individual and the type of Ad. To begin to address this possibility, the gp19K gene from 17 very diverse Ad7 (subgroup B) clinical isolates was amplified by the polymerase chain reaction, and the DNA sequences were determined. The Ad7 gp19K sequence was 98% identical to that of Ad3. Surprisingly, we found complete conservation of the amino acid sequence of gp19K from all but one of the clinical isolates; one isolate had a conservative Ala to Val substitution. Gp19K from Ad7 clinical isolates representing distinct Ad7 genotypes co-immunoprecipitated with class I antigens. Our data indicate that there is very strong evolutionary pressure to maintain the sequence of gp19K in Ad7. The only known function for gp19K from different Ad serotypes is binding to class I antigens. It is interesting to consider, therefore, what selective pressure operates to maintain the sequence of gp19K among serotypes within a subgroup, and yet allows for very significant divergence in the sequence of gp19K among serotypes in different subgroups. The possible role of MHC class I antigens in this selection process is discussed.

Nosocomial transmission of adenovirus type 3 associated with fatalities in infants has not been frequently reported. This report describes the nosocomial spread of adenovirus types 2 and 3 among infants with bronchopulmonary dysplasia in a chronic (transitional) care facility. The index case developed pneumonia with a clinical deterioration in respiratory status 8 days after admission. Within the next 10 to 30 days 9 other infants and 2 health care personnel became ill with respiratory symptoms. Three of these 10 infants had progressive respiratory failure and 2 of them died. All of these infants had underlying chronic lung disease of bronchopulmonary dysplasia. The overall attack rate was 30% (10 of 33). Further spread of adenovirus was prevented by using barrier precautions and masks while performing tracheostomy care. Adenovirus isolates were serotyped as Ad3 in 4 patients and 1 staff member, as Ad2 in 3 patients, and as a combination of Ad2 and Ad3 in 1 patient. Two fatalities were associated with Ad3 infection. Three isolates from 2 patients and 1 staff member were not available for typing. Restriction endonuclease analysis was performed on all of these isolates of Ad3 and Ad2. There was no genetic heterogeneity in the isolates, suggesting a common source.

Objective: To study patterns of transmission of epidemic keratoconjunctivitis (EKC) in a chronic care facility and to assess control measures and prevent future outbreaks in this setting., Design: A retrospective cohort study., Setting: A 120-bed, four-unit, skilled nursing facility., Patients: Residents and employees of the above facility., Interventions: Increased frequency of cleaning; use of bleach disinfectant; universal precautions in handling eye secretions from residents with conjunctivitis; cohorting residents by unit; suspension of new admissions; closure of common gathering areas., Measurements: Resident demographics; possible risk factors for infection among residents (including mobility, underlying illness, medications, involvement in social activity, level of confusion) and among employees (including co-morbid illnesses and eye conditions, exposures to persons with conjunctivitis, visits to eye care specialists, use of contact lenses or glasses); testing of conjunctival specimens from symptomatic persons for viral and bacterial agents., Results: Of 95 residents on three chronic care units, 47 (attack rate 49%) had onset of eye symptoms consistent with EKC between September 14 and December 7, 1990. Thirty-eight (81%) of these had onset following the onset of symptoms in a resident with dementia who, despite habitual eye-rubbing and wandering into other residents' rooms, was not isolated or restricted in any way. Attack rates were higher (though not statistically significant) among more mobile residents (60% for ambulatory residents) and among those considered by staff to be confused (56%). Rapid antigen detection and culture confirmed adenovirus type 37 as the etiologic agent., Conclusions: Transmission of infection with adenovirus type 37 was successfully interrupted following strict infection control, suspension of new admissions, cohorting of residents by unit, and change to a disinfectant that inactivates adenovirus. Recognition of conjunctivitis as an appropriate reason for restricting movement of an infected resident may have prevented extensive viral transmission in this outbreak.

Adenoviruses are gradually being recognized as a significant source of morbidity and mortality in the immunocompromised patient population. We report a bone marrow transplant patient who developed severe abdominal pain accompanied by marked elevations in serum pancreatic and hepatic enzyme levels. She died shortly thereafter. Autopsy revealed hemorrhagic pancreatitis and fulminant hepatic necrosis. Both the pancreas and liver contained intranuclear inclusions consistent with adenovirus; electron microscopy confirmed that there were, indeed, adenoviral particles. This report of adenoviral pancreatitis emphasizes the diversity of manifestations seen with adenoviral infection.

The polymerase chain reaction (PCR) was used to detect adenovirus DNA in formalin-fixed, paraffin-embedded tissue obtained post mortem. Adenovirus DNA was successfully amplified from specimens of lung and liver from two patients with disseminated adenovirus infection confirmed by virus isolation, electron microscopy and/or immunohistochemistry. Negative results were obtained for specimens of lung from two patients with cytomegalovirus pneumonia. The specificity of the adenovirus PCR was confirmed by means of a digoxigenin-labelled probe generated in a separate PCR. Detection of viral nucleic acid by PCR in tissues obtained post mortem has considerable diagnostic potential.

We describe a fatal case of adenovirus pneumonia accompanied by encephalitis in a neonate who showed lethargy on the 6th day and died on the 12th day. Adenoviral particles as well as viral intranuclear inclusions were noted in pulmonary alveolar epithelium cells. Neuropathological examination revealed diffuse oedema, perivascular cuffing and gliosis in the white matter. Adenovirus type 11 was isolated from lung, hilar lymph node, and brain tissue. This is the first instance of adenovirus isolation from brain tissue in a newborn infant. The virological and neuropathological findings suggest the invasion of neural tissue by adenovirus and substantiate the significance of neurological symptoms observed in neonatal adenovirus infection.

Monoclonal antibody (MAb) preparations specific for the enteric adenoviruses of subgenus F (AdF) were generated and evaluated as typing reagents in virus neutralization tests and enzyme-linked immunosorbent assays (ELISAs). A panel of 11 genome types of adenovirus 40 (Ad40), 24 genome types of Ad41, and 47 adenovirus prototype strains was used to determine the specificities of the MAbs in the two assays. In this way two MAbs, MAb 40-1 (anti-Ad40) and MAb 41-1 (anti-Ad41) were selected. These two MAbs showed strict type specificity in both assays. A third MAb reacted in an ELISA with all 47 human adenovirus types. With two other MAbs, three antigenic subtypes of Ad41 could be distinguished by their reactivities in virus neutralization tests and ELISAs. On the basis of the five selected MAbs, a sensitive ELISA system was developed for the direct detection and simultaneous typing and subtyping of Ad40 and Ad41 present in stool specimens. The five MAbs were also used to study the epidemiology of infections with Ad40 and Ad41 in The Netherlands in the period 1981 through 1989. It was shown that there were no significant fluctuations in the annual incidence of the cluster of enteric adenoviruses as a whole. This cluster should therefore be considered to belong to the "endemic" rather than the "epidemic" adenoviruses. The relative incidence of Ad40 infections compared with that of Ad41 infections changed considerably during the period studied; the proportion of Ad41 infections rose from about 30% in 1981 to about 95% in 1986, after which it stabilized at 90 to 95%. The proportion of one of the subtypes of Ad41 (Ad41 subtype M3) increased from about 40 to 80% in the same period.

We report a 5 year prospective study of episodes of rotavirus, subgenus F adenovirus, and astrovirus gastroenteritis diagnosed by electron or immune electron microscopy in a single regional virology laboratory. Of 1426 total infections, the numbers in each category were 1117 (78.3%), 254 (17.8%), and 20 (7.9%), respectively. Using restriction endonuclease analysis or immune electron microscopy, all but 20 of the subgenus F adenovirus strains were classified as type 40 (n = 50) or type 41 (n = 184). Rotavirus and astrovirus infections were more prevalent in winter than summer, whereas subgenus F (either type 40 and 41) adenoviruses showed no seasonal variation in prevalence. The ratio of type 40 to all typable subgenus F adenoviruses declined between 1984 and 1986 and then increased again. Adenoviruses were relatively more important as causes of viral gastroenteritis in infants aged less than 6 months than in toddlers aged 12 months or more, but even in young infants more rotavirus than adenovirus infections were diagnosed. Our data confirmed the epidemiological differences between rotavirus, subgenus F adenovirus and rotavirus gastroenteritis and documented the shared epidemiological characteristics of type 40 or 41 adenovirus infections.

57 adenoviruses (AV's) isolated from 1979 through 1986 from about 200 patients with (kerato) conjunctivitis were identified as AV8, 7, 3, 17/H29, 9, 2, and 10, in decreasing order. DNA restriction analysis with 5 enzymes revealed the three strains of the intermediate 17/H29 strains to be identical. The same conclusion was reached for the two AV9 strains, but the latter differed from the AV9 prototype in two enzymes. Restriction site alterations were physically mapped for 3 enzymes and for an AV8 variant differing in HindIII.

Two divergent strains of adenovirus type 31 were analyzed by neutralization test and restriction endonuclease (RE) patterns in an effort to find the basis for their genetic variability. One strain, isolated from the throat of a child in Maryland during an upper respiratory illness in 1968, was partially neutralized by Ad 31 antisera (to 16-fold lower than homologous titer) while its own antiserum fully neutralized prototype Ad 31 virus, but shared only 9% of comigrating RE fragments with Ad 31 prototype (vs. 30% with Ad 18 prototype); however, PCR tests specific for the inverted terminal repeat (ITR) sequence of Ads 12 and 18 were negative. The other strain, recovered from a stool sample from an infant with diarrhea in Georgia in 1979, was inhibited by Ad 31 antiserum to within 4-fold homologous titer, but shared only 15% of comigrating fragments with Ad 31 prototype (vs. 91% with Ad 18 prototype); ITR-specific PCR tests with this virus were positive for Ad 12/Ad 18. These data suggest that both strains are from separate evolutionary lines of Ad 31 unrelated to all other isolates studied to date by RE analysis, and that the partial neutralization by prototype Ad 31 antisera might represent small mutations in the hexon gene.

'Adenovirus keratoconjunctivitis' is a diagnosis commonly made in Eye Casualty Departments. There are no clinical features that allow identification of the serotype causing the infection. The changeable patterns of presentation and manifestation of the different serotypes and the capacity of adenovirus to mimic other viral or bacterial infections is an enigma. Adenovirus infections are usually self-limiting; permanent visual loss is very rare. The use of corticosteroids is generally limited to cases with severe symptoms such as glare and incapacitating blurring of vision. Corticosteroids will not alter the basic pathogenesis of the disease but will suppress inflammatory signs. The sub-epithelial lesions are an immunopathological phenomenon resulting from the interaction of viral antigen and antibody in the anterior stroma with the cornea acting as a 'blotter'. An effective antiviral remains elusive.

Two new adenovirus (AV) subgenus D serotypes, which we propose as AV candidates 48 and 49, are described. Both AV 48 and 49 were unique by serum neutralization and the pattern of restriction endonuclease fragments. By the hemagglutination inhibition assay there was a significant reciprocal cross between AV 48 and 44 and one-way crosses between AV 48 and AV 37 sera and between AV 30 and AV 49 sera. Eleven AV 48 and 5 AV 49 isolates, mainly from AIDS patients, have been identified at the Viral and Rickettsial Disease Laboratory.

Genome analysis was carried out on 74 adenovirus 4 (Ad4) isolates from patients in Manchester between 1984 and 1989. Most of the isolates were associated with conjunctivitis. Of the 74 isolates studied, 51 were Ad4a and 10 were Ad4p (the prototype strain). The remaining isolates consisted of two new genome types we have designated Ad4a2 (10 isolates) and Ad4a3 (3 isolates). Most of the genome types co-circulated during the period of study. The Bst E II and Xho I restriction maps of the new variants are presented and compared with those of Ad4p. We are unable to associate genome types with particular clinical presentations.

Adenoviruses isolated over a period of 10 years from patients with conjunctivitis were typed by neutralisation test using reference sera and by restriction endonuclease fragment (REF) analysis. Adenoviruses were isolated from 516 of 10,232 patients tested (5.0%); 154 were identified as type 3, 153 as type 4, 70 as type 7, 17 as type 10 and 122 as other types. At any one time, several serotypes co-circulated. The prevalence of types varied. Type 4 was not isolated in the first 2 years and then gradually increased in incidence, becoming the most frequently isolated type after 1987. Two periods of increased isolation frequency occurred: firstly from May to August 1981, when 8-28% of patients per month were found to be adenovirus-positive, with serotype 3 being predominant; and secondly from January 1989 to August 1990, when 8-20% of patients per month were adenovirus-positive, predominantly with type 4. Analysis of REFs showed that several different genotypes exist within serotypes. These also co-circulated over long periods with intermittent changes of the predominant genotype. The prototype strain Ad3GB was isolated more frequently than five other Ad3 genotypes from 1981 to 1988, after which a variant strain Ad3a was most common.

Objective: To investigate the possible relationship between enteric adenovirus types 40 and 41 and intestinal intussusception in children., Design: Prospective, case-control patient study., Patients: Sixty-three consecutive children suspected clinically of having intestinal intussusception were enrolled in this study. Of these, 25 children (mean age, 1.4 years; range, 3 months to 5 years) had barium enema examination-proved intussusception. Age-matched normal controls (24) and controls with diarrhea (21) were obtained within 1 month of the index case., Measurements and Results: Stools were tested for the presence of nonenteric adenovirus and enteric adenovirus using a monoclonal antibody-based enzyme immunoassay. Five (20%) of 25 children with intussusception had nonenteric adenovirus in their stools compared with one (4%) of 24 normal controls, none (0%) of 21 of the controls with diarrhea, and none (0%) of 37 patients suspected of having intussusception who had negative results on barium enema examination. However, no stool samples were positive for enteric adenovirus., Conclusions: Nonenteric adenovirus infection and intestinal intussusception may be associated. However, because enteric adenovirus was not found in any of the groups studied, no conclusions can be made regarding their possible influence on the risk for developing intussusception.

Four consecutive epidemics of keratoconjunctivitis caused by adenovirus 8 (Ad8) occurred over a 5-year period in Brest, France. A selection of 30 strains isolated during this period was studied by DNA restriction enzyme analysis using nine restriction enzymes. BglI and SacI were the most discriminative enzymes and allowed the recognition of four DNA variants, all different from the prototype strain Trim. Within each of the epidemics, the strains tested could not be distinguished in this analysis. Between strains from different epidemics differences in DNA structure could be detected however. Thus, the Ad8 epidemics of 1983/1984, 1984, 1987, and 1988 appear to have been due to DNA variants Ad8/D7, D8, D9, and D10, respectively. These results demonstrate that the DNA of Ad8 seems to display a considerable variability, comparable to that observed with Ad7 and Ad21. As has been described for Ad7, Ad21 and Ad41, successive DNA variants of Ad8 prevail during one or more years, and are then replaced by other, newly emerging variants sometimes associated with epidemics.

Adenovirus infections commonly occur in childhood and produce a wide range of clinical disease. The most common sites of infection are the respiratory and gastrointestinal tracts but involvement of cardiovascular, neurological, cutaneous, ophthalmic, renal, and hepatic systems can also occur. A case of toxic shock-like syndrome with symptoms of multiorgan involvement resulting from adenovirus infection is reported.

In order to detect rotavirus and adenovirus 557 feces samples from hospitalized children (0-5 years of age) were analysed from June 1987 to July 1990 in Goiânia city. Two hundred and ninety one samples were from children with diarrhoea and 266 were from children without diarrhoea. Amongst this later group, 64 samples were from children from the nursery. Two hundred and sixty one out of 557 samples were analysed by immunoelectron microscopy (IEM), polyacrylamide gel electrophoresis (SDS-PAGE) and enzymatic immunoassay for rotavirus and adenovirus (EIARA) whereas the rest (296 samples) were analysed by SDS-PAGE and EIARA. Positivity of rotavirus and adenovirus was 17.2% and 2.1% respectively. Concerning rotavirus and adenovirus there was 29.2% and 2.4% positivity within the group with diarrhoea and 4.1% and 1.5% positivity amongst children without diarrhoea (p < 0.05). Rotavirus were more prevalent amongst children which age ranged from 1 to 11 months of age. No newborn child from the nursery was positive for rotavirus. Adenovirus were detected amongst children from 1 to 3 years of age. Rotavirus circulation peak occurred between May and August (p < 0.05) and no positive case was detected from December to February. Two hundred out of 291 diarrheic samples were also studied concerning bacteria and pathogenic parasites and equal percentages (17.0%) were found for both rotavirus and pathogenic bacteria. Eighty nine samples of rotavirus were detected by SDS-PAGE and 86 of these (96.6%) belonged to the subgroup II with 13 different electrophoretic patterns. Predominance of a given electrophoretic profile was observed in each year of the study.

Restriction endonuclease analysis using 10 restriction enzymes was performed on six and three wild isolates of adenovirus (Ad) type 12 and 18, respectively. Among the Ad12 strains, five DNA variants could be identified. The degree of pairwise comigration of restriction fragments suggests a high degree of genomic diversity within Ad12. The wild isolates of Ad18, on the other hand, displayed a low degree of genetic variability and comprised one DNA variant closely related to the prototype strain. The BglII, BstEII, and HindIII restriction endonuclease patterns of Ad18 were inconsistent with those originally presented. Identical RE-patterns among Ad18 prototype strains (DC) obtained from four different sources, including directly from the American Type Culture Collection, verify that the genuine Ad18 prototype was used in the present study. Using the revised restriction patterns of BglII, BstEII, and HindIII, a proper identification of Ad18 will be facilitated.

The use of the polymerase chain reaction (PCR) for detection of human adenoviruses in diluted stool samples was investigated. Two sets of nested primers, including primers specific for the hexon-coding region and for the E1B region of enteric adenoviruses (EAd), were assessed by two-step amplification. The primers constitute two different PCR systems designed for the detection of adenoviruses belonging to all six subgenera (A-F), and the two EAds Ad40 and Ad41, respectively. In a two-step PCR mediated amplification a single virus particle was detected when the two sets of general hexon primers or EAd specific primers were used. Earlier results from PCR detection of adenoviruses in stool from children suffering from diarrhea gave indications that adenovirus particles are commonly shed in stools without being identified as the cause of illness [Allard et al.: Journal of Clinical Microbiology 28:2659-2667, 1990]. Therefore, the general and the EAd specific PCR assays were assessed on 150 stool specimens from three groups including 50 healthy children, 50 healthy adults, and 50 adults suffering from diarrhea. When the two sets of general hexon primers were used, 25 of the 50 specimens from the healthy children (mean age 21 months) were found positive by two-step PCR amplification. Nine of the 50 specimens from the healthy adults (mean age 32 years) were found positive whereas 12 of the 50 specimens from sick adults (mean age 31 years) gave amplification products, using the two sets of general hexon primers in a nested fashion. None of the 150 specimens were found to be positive by two-step PCR amplification using the two sets of EAd-specific primers.(ABSTRACT TRUNCATED AT 250 WORDS)

An adenovirus (Ad) type 8 outbreak was prospectively studied in a neonatal intensive care unit. Nasopharyngeal secretions were cultured weekly for viruses and clinical information was obtained daily in each infant. Eleven of 112 neonates were infected with Ad; 8 of 9 available isolates represented one variant of Ad 8. The median age at infection was 54 days and the median duration of virus shedding was 2 days. Seven of 11 infants had onset of new symptoms and/or required acute respiratory support; only 2 infants had eye disease. Maternal characteristics, race, gender, age at entry into study and respiratory distress syndrome at birth were similar for both groups. Ad-infected infants tended to have earlier gestations and lower birth weights. Ad-infected neonates stayed longer in the neonatal intensive care unit, required more days of respiratory support and were more likely to develop bronchopulmonary dysplasia. Thus an association was established between lung disease in premature neonates and Ad infection.

The fastidious enteric adenovirus (FEAd) types 40 (Ad40) and 41 (Ad41) are found in stool specimens of infants and young children in association with gastroenteritis. Although they can be isolated routinely from clinical specimens by using 293 cells, they are propagated with variable success in cell lines which support the replication of other adenovirus serotypes. HeLa cells are generally considered to be nonpermissive for the replication of FEAds, but in this study, Ad40 and Ad41 grew to comparable titers in individual 293 and HeLa cells. However, virus was not efficiently released from infected HeLa cells and thus did not undergo multiple cycles of infection in HeLa cell cultures. The block in virus release was not overcome in KB18 cells which, like 293 cells, constitutively express proteins encoded by the E1B region of a subgroup C adenovirus (in this case Ad2). Moreover, it was apparent from these studies that Ad40 and Ad41 have particle-to-infectious unit ratios several orders of magnitude greater than that for Ad5, even in 293 cells which express the E1A and E1B proteins of Ad5 and are considered to be permissive for replication of the FEAds. Neither the block in release of progeny virus nor the high particle-to-infectious unit ratio is explained solely by the defect in expression of the E1B 55K protein identified by Mautner et al. (V. Mautner, N. MacKay, and V. Steinthorsdottir, Virology 171:619-622, 1989; V. Mautner, N. MacKay, and K. Morris, Virology 179:129-138, 1990).

Objective: Human enteric adenovirus (EAd) types 40 and 41 cause diarrhea in young children, but little is known about their association with outbreaks of diarrhea in the child care setting. This study evaluated EAd as a cause of outbreaks of diarrhea among infants and toddlers in day care centers., Design: Stool specimens were collected weekly regardless of symptoms during four periods from January 1986 to April 1991, from children 6 to 24 months of age enrolled in prospective studies of diarrhea in day care centers. All diarrhea stool specimens were tested for bacterial enteropathogens, rotavirus, and Giardia lamblia. A total of 131 outbreaks occurred during the study. No etiologic agent was identified in 77 outbreaks. Stool specimens from 75 of these 77 outbreaks and from another 21 outbreaks of diarrhea with a known cause were evaluated for EAd with a monoclonal antibody-based enzyme immunoassay., Results: A total of 4402 stool specimens from 613 children from these 96 outbreaks was tested for EAd. The virus was detected in specimens collected during 10 outbreaks, 3 of which occurred in 1986, 3 in 1988, 1 in 1989, 1 in 1990, and 2 in 1991. Of 249 children, 94 (38%) in these 10 EAd outbreaks were infected with EAd. In 51 children (54%) the infection was symptomatic and in 43 (46%) it was asymptomatic. Outbreaks lasted 7 to 44 days (mean 24.5 days). Duration of EAd excretion ranged from 1 to 14 days (mean 3.9 days), with excretion occurring from 7 days (mean 2.6) before diarrhea began to 11 days (mean 5.3 days) after diarrhea stopped., Conclusion: Enteric adenovirus types 40 and 41 are an important cause of outbreaks of diarrhea among children attending day care centers, often involve children in more than one room, and frequently produce asymptomatic infection.

DNA restriction analysis was carried out on a sample of 73 adenovirus strains isolated in Buenos Aires from nasopharyngeal aspirates of children with lower acute respiratory infection between 1984 and 1988. Thirty-five isolates (47.9%) were classified as members of subgenus B. Of these, three were identified as a new genome type of Ad3p denominated Ad3p3; five strains corresponded to genome type 7b and two to genome type 7c. The other 25 isolates were identified as the recently recognized genome type 7h. All 6 fatalities recorded within this group of 73 children were associated with infection by Adenovirus genome type 7h. Thirty-seven isolates (50.7%) were classified within subgenus C that corresponded to 9 different genome types denominated 1p (n = 5); 1# (n = 2); 2p (n = 4); 2b (n = 6); 2# (n = 5); 5# (n = 4); 5* (n = 7) and 5+ (n = 2). All genome types of subgenus C were compared with the data reported by Adrian et al. (Archives of Virology 112:235-238, 1990). The Ad1p and Ad1# genome types could be allocated to AV1 genome types D1 and D10, respectively. Ad2b genome type could be allocated to AV2 genome type D25. No counterparts were found for the remaining 6 genomic variants. Only one isolate was identified as Ad4a of subgenus E. The comparison of the results of the present study with those of the molecular characterization of Chilean strains isolated between 1984 and 1987 suggests that the adenovirus strains associated with respiratory disease of children may be common in both countries.

Adenovirus was cultured from various gastrointestinal sites and one lung specimen from seven immunosuppressed patients, four of whom were bone marrow transplant recipients. Histologic examination and in situ hybridization studies of the lung demonstrated adenovirus in the bronchial epithelium and alveolar lining cells. In contrast, none of the 34 gastrointestinal biopsies performed within 30 days of the positive adenovirus culture showed histologic or molecular evidence of invasive adenovirus in the gastrointestinal mucosa. These results suggest that isolation of adenovirus from gastrointestinal biopsy specimens taken from immunosuppressed patients probably does not indicate an invasive adenoviral infection.

It is now well known that several viruses are responsible for acute diarrhoea or gastroenteritis in both children and adults. These viruses are difficult to identify since most of them cannot be isolated by stool cultures on cells. The reality of proven reinfection by some of these organisms is not always clearly understood, even though the existence of several serotypes in the same group (notably rotavirus) can be blamed, and this explains why vaccines are difficult to develop.

Formation of progeny viruses in the nuclei of HeLa cells infected with adenovirus type 5 was studied at the ultrastructural level by in situ hybridization techniques allowing specific detection of either viral double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA). Prior to the initiation of replication of viral genomes, infective DNA molecules which entered the nucleus of the target cell were randomly distributed among host chromatin fibers including nucleolus-associated chromatin. They were double-stranded, that is, without single-strand breaks. Such association of viral DNA with host condensed chromatin also occurred in mitosis. The initiation of viral genome replication occurred simultaneously with the appearance in the nucleoplasm of small fibrillar regions containing intermingled viral dsDNA and ssDNA. Later, at the intermediate stage of nuclear transformation, viral dsDNA and ssDNA molecules were almost entirely separated into two contiguous substructures. At this stage, viruses were observed occasionally in the vicinity of viral ssDNA accumulation sites. Still later, an additional substructure developed in the centre of the nucleus which consisted of large quantities of viral dsDNA, traces of viral ssDNA and abundant viruses. Portions of viral ssDNA were attached to some viruses even at late stage of nuclear transformation, an association which strongly suggests the occurrence of encapsidation of at least some of the viral genomes while they are still engaged in replication.

Three hundred and eighty-five children without a diagnosis of diarrhoeal illness had faecal specimens collected in order to detect rotavirus infection. The survey was conducted in Goiânia city--central Brazil and the children aged 0-11 years old were recruited (268 in day care centers and 117 from a local outpatient clinic--Hospital Lucio Rebelo). Detection of rotavirus was done by polyacrylamide gel electrophoresis (PAGE-SDS), and 89 specimens were also analyzed by commercial enzyme immunoassays for rotavirus and adenovirus (EIARA). Rotaviruses and adenoviruses were only detected among the children attending the outpatient clinic. Prevalence rates of rotavirus and adenovirus excretion were of 1.7% and 1.6%, respectively. Both viruses were found among children from 1 to 2 years old.

Among a group of hospitalized children in Fukuoka, southern Japan, an epidemic of pharyngoconjunctival fever-like disease caused by adenovirus type 11 was observed in the autumn of 1988. Of the 47 children studied 38 were seronegative in neutralizing antibody for adenovirus type 11 before the epidemic, and seroconversion occurred in 16 (42%) including 5 subclinical cases. Of the 11 symptomatic patients the incidences of conjunctivitis, pharyngitis and fever were 91, 64 and 46%, respectively. Four patients (36%) had all three symptoms. Fifteen patients (94%) were boys. The sex predominance and high incidence of conjunctivitis suggested that infection may have been transmitted in the large bathroom where boys took baths together.

Adenovirus (Ad) type 40 and 41 DNAs were directly extracted from stool specimens of children with gastroenteritis. Two new strains of Ad41, Sanekata and Ehime strain, were cloned and their restriction maps were constructed. The left terminal end of the cloned Ad41 genome, EcoRI-E fragment of the Sanekata strain and EcoRI-F fragment of the Ehime strain, had transforming ability in rat 3Y1 cells. Only one of the 35 isolates of Ad40 tested showed a different restriction profile, while three different restriction profiles were found in DNAs from Ad41 isolates.

A large outbreak of epidemic keratoconjunctivitis occurred in Alice Springs, Australia, between July and September 1989. Initially, there was a large outbreak within two child care centers, followed by a more prolonged outbreak within the community. A simultaneous outbreak occurred among staff at the local hospital. Infected children developed mild conjunctivitis, whereas adults developed typical keratoconjunctivitis. Conjunctival swabs from 30 cases were cultured and adenovirus was recovered from 18, all of which were typed by neutralization assay as adenovirus type 8. DNA restriction fragment analysis of the genomes of 5 isolates revealed they were identical, distinct from prototype adenovirus type 8, and nearly identical to adenovirus genome type 8C (Ad-8C), which was previously reported in the southern United States in the early 1970s. Ad-8C was present in another region of Australia greater than or equal to 1 year before the epidemic.

An analysis of adenovirus strains isolated in the Soviet Union from 1976 to 1988 revealed four genome types of adenovirus type 3 (Ad3), i.e., Ad3a4, Ad3a9, Ad3a10, and Ad3a11, and four genome types of adenovirus type 7 (Ad7), i.e., Ad7p, Ad7a, Ad7a(1-5), and Ad7f1, identified with the DNA restriction enzymes BamHI, BglII, and HindIII. Three of them, Ad3a10, Ad3a11, and Ad7f1, are newly discovered. The genetic heterogeneity of adenoviruses was examined with restriction endonuclease Cfr13I with a 4-base recognition cleavage site. Eighteen different restriction patterns were identified among 21 selected Ad3 strains after cleavage of DNA with Cfr13I. Eight different subtypes were identified among 20 Ad7 strains by the same technique. For estimation of the relationships among these genome subtypes, pairwise analyses of comigrating DNA restriction fragments from isolates of Ad3 and Ad7 were done after digestion with Cfr13I or with restriction endonucleases recognizing DNA sequences of 6 bp. Surprisingly, the results were very discrepant.

The relative importance of astrovirus and adenoviruses as etiologic agents of diarrhea among children in day care was examined. Stool specimens from this prospective study were screened for both astrovirus and adenovirus hexon with two new indirect double-antibody assays and for enteric adenoviruses with an EIA specific for serotypes 40 and 41. Astrovirus was detected in a significantly greater percentage of children with diarrhea (4%, 21/524) than of those without (less than 1%, 1/138) (P less than .05); however, no difference between such such children with adenovirus infections was found (8%, 43/565, and 8%, 10/129, respectively). Overall, 30% (13/43) of all adenovirus hexon-positive specimens were enteric serotypes, and by extrapolation, enteric adenoviruses were identified in an equal percentage of children (2%) with and without diarrhea. This study documents the presence of astrovirus and enteric adenoviruses among children in day care in the United States, associates astrovirus with diarrhea in this setting, and suggests that viral agents may be the most common enteric pathogens among children with diarrhea in day care.