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In 2018, a cluster of pediatric human parechovirus (HPeV) infections in 2 neighboring German hospitals was detected. Viral protein 1 sequence analysis demonstrated co-circulation of different HPeV-3 sublineages and of HPeV-1 and -5 strains, thereby excluding a nosocomial outbreak. Our findings underline the need for HPeV diagnostics and sequence analysis for outbreak investigations.

Recent parechovirus A3 (PeV-A3) outbreaks in Australia suggest lower population immunity compared with regions that have endemic PeV-A3 circulation. A serosurvey among populations in the Netherlands, the United States, and Australia before and after the 2013 Australia outbreak showed high PeV-A3 neutralizing antibody prevalence across all regions and time periods, indicating widespread circulation.

We describe a lethal respiratory outbreak among wild chimpanzees in Uganda in 2013 for which molecular and epidemiologic analyses implicate human rhinovirus C as the cause. Postmortem samples from an infant chimpanzee yielded near-complete genome sequences throughout the respiratory tract; other pathogens were absent. Epidemiologic modeling estimated the basic reproductive number (R0) for the epidemic as 1.83, consistent with the common cold in humans. Genotyping of 41 chimpanzees and examination of 24 published chimpanzee genomes from subspecies across Africa showed universal homozygosity for the cadherin-related family member 3 CDHR3-Y529 allele, which increases risk for rhinovirus C infection and asthma in human children. These results indicate that chimpanzees exhibit a species-wide genetic susceptibility to rhinovirus C and that this virus, heretofore considered a uniquely human pathogen, can cross primate species barriers and threatens wild apes. We advocate engineering interventions and prevention strategies for rhinovirus infections for both humans and wild apes.

We identified new clinical manifestations associated with Senecavirus A infection in neonatal piglets in Brazil in 2015. Immunohistochemical and molecular findings confirmed the association of Senecavirus A with these unusual clinical signs and more deaths. Other possible disease agents investigated were not associated with these illnesses.

Senecavirus A has been infrequently associated with vesicular disease in swine since 1988. However, clinical disease has not been reproduced after experimental infection with this virus. We report vesicular disease in 9-week-old pigs after Sencavirus A infection by the intranasal route under experimental conditions.

The infectious etiology of myocarditis often remains unidentified. We report a case of myocarditis associated with human parechovirus (HPeV) infection in an adult. HPeV is an emerging pathogen that can cause serious illness, including myocarditis, in adults. Testing for HPeV should be considered in differential diagnosis of myocarditis.

HPeV should be tested for in all young children suspected to have HPeV or enterovirus infection., Human parechoviruses (HPeVs) often cause severe illness among young children. National surveillance with routine testing of all cerebrospinal fluid, fecal, and tissue samples was conducted during January 2009–December 2012 in all counties in Denmark (6,817 samples from 4,804 children were screened for HPeV). We detected HPeV RNA in 202 (3.0%) specimens from 149 persons. Young infants were at highest risk for HPeV, and 9 (6%) of the HPeV-infected children died, probably of their HPeV illness. HPeV3 was the most common genotype identified, and 5 closely related clades of HPeV3 circulated in Denmark throughout the study period. Our study adds perspective on the prevalence and clinical and molecular virologic characteristics of HPeV infection.

In 2014, sepsis-like illness affected 9 full-term newborns in 1 hospital in Austria. Although results of initial microbiological testing were negative, electron microscopy identified picornavirus. Archived serum samples and feces obtained after discharge were positive by PCR for human parechovirus 3. This infection should be included in differential diagnoses of sepsis-like illness in newborns.

We assessed Aichi virus shedding in patients with gastroenteritis and negative test results for other viral and bacterial infections. High concentrations of up to 1.32 × 1012 RNA copies/g stool were found in 10 (2.0%) of 499 outpatients sampled in northern Germany, 2004. These data substantiate Aichi virus pathogenicity in humans.

To identify a candidate etiologic agent for turkey viral hepatitis, we analyzed samples from diseased turkey poults from 8 commercial flocks in California, USA, that were collected during 2008–2010. High-throughput pyrosequencing of RNA from livers of poults with turkey viral hepatitis (TVH) revealed picornavirus sequences. Subsequent cloning of the ≈9-kb genome showed an organization similar to that of picornaviruses with conservation of motifs within the P1, P2, and P3 genome regions, but also unique features, including a 1.2-kb sequence of unknown function at the junction of P1 and P2 regions. Real-time PCR confirmed viral RNA in liver, bile, intestine, serum, and cloacal swab specimens from diseased poults. Analysis of liver by in situ hybridization with viral probes and immunohistochemical testing of serum demonstrated viral nucleic acid and protein in livers of diseased poults. Molecular, anatomic, and immunologic evidence suggests that TVH is caused by a novel picornavirus, tentatively named turkey hepatitis virus.

To investigate a possible role for human rhinovirus C in respiratory exacerbations of children with cystic fibrosis, we conducted microbiologic testing on respiratory specimens from 103 such patients in São Paulo, Brazil, during 2006–2007. A significant association was found between the presence of human rhinovirus C and respiratory exacerbations.

Increased genomic diversity of these viruses is demonstrated., Rhinoviruses and enteroviruses are leading causes of respiratory infections. To evaluate genotypic diversity and identify forces shaping picornavirus evolution, we screened persons with respiratory illnesses by using rhinovirus-specific or generic real-time PCR assays. We then sequenced the 5′ untranslated region, capsid protein VP1, and protease precursor 3CD regions of virus-positive samples. Subsequent phylogenetic analysis identified the large genotypic diversity of rhinoviruses circulating in humans. We identified and completed the genome sequence of a new enterovirus genotype associated with respiratory symptoms and acute otitis media, confirming the close relationship between rhinoviruses and enteroviruses and the need to detect both viruses in respiratory specimens. Finally, we identified recombinants among circulating rhinoviruses and mapped their recombination sites, thereby demonstrating that rhinoviruses can recombine in their natural host. This study clarifies the diversity and explains the reasons for evolution of these viruses.

Using a simple metagenomic approach, we identified a divergent human parechovirus (HPeV) in the stool of a child in Pakistan. Genomic characterization showed this virus was distinct enough from reported HPeV types to qualify as candidate prototype for the seventh HPeV type.

Human parechoviruses (HPeVs) were detected by reverse transcription–PCR in 16.1% of 335 stool samples from children

To determine links between human rhinoviruses (HRV) and asthma, we used data from a case-control study, March 2003-February 2004, among children with asthma. Molecular characterization identified several likely new HRVs and showed that association with asthma exacerbations was largely driven by HRV-A and a phylogenetically distinct clade of 8 strains, genogroup C.

Global surveillance for a novel rhinovirus genotype indicated its association with community outbreaks and pediatric respiratory disease in Africa, Asia, Australia, Europe, and North America. Molecular dating indicates that these viruses have been circulating for at least 250 years.

Identification of HPeV-6 will advance HPeV diagnosis and epidemiology., Using Vero cells, we isolated a virus (NII561-2000) from a cerebrospinal fluid specimen of a 1-year-old girl with Reye syndrome. The determined amino acid sequence of the virus indicated that the isolate was a human parechovirus (HPeV), a member of Picornaviridae. Neutralization test showed that the NII561-2000 virus had distinct antigenicity to HPeV-1, HPeV-2, and HPeV-3, and that the sequence was distinct from these types as well as from HPeV-4 and HPeV-5. Thus, we propose the virus (NII561-2000) as the prototype of HPeV-6. We isolated 10 NII561-2000–related viruses, 14 HPeV-1, 16 HPeV-3, and 1 HPeV-4 of 41 HPeVs from various clinical samples collected in Niigata, Japan. Clinical symptoms of the persons infected with the NII561-2000–related viruses were infectious gastroenteritis, rash, upper respiratory tract infection, and paralysis, in addition to Reye syndrome in the 1-year-old girl.

Maternal antibodies may protect infants from severe illness caused by this pathogen., Human parechovirus type 3 (HPeV3) is an emerging pathogen that causes sepsis and meningoencephalitis in young infants. To test the hypothesis that maternal antibodies can protect this population, we measured neutralizing antibody titers (NATs) to HPeV3 and other genotypes (HPeV1 and HPeV6) in 175 cord blood samples in Japan. The seropositivity rate (>1:32) for HPeV3 was 61%, similar to that for the other genotypes, but decreased significantly as maternal age increased (p1:512) until age 6 months. All intravenous immunoglobulin samples tested elicited high NATs to HPeV3. Our findings indicate that maternal antibodies to HPeV3 may help protect young infants from severe diseases related to HPeV3 and that antibody supplementation may benefit these patients.

Syndromic surveillance was useful for outbreak monitoring, and public health response helped reduce hospitalization times., From October 2013 through February 2014, human parechovirus genotype 3 infection was identified in 183 infants in New South Wales, Australia. Of those infants, 57% were male and 95% required hospitalization. Common signs and symptoms were fever >38°C (86%), irritability (80%), tachycardia (68%), and rash (62%). Compared with affected infants in the Northern Hemisphere, infants in New South Wales were slightly older, both sexes were affected more equally, and rash occurred with considerably higher frequency. The New South Wales syndromic surveillance system, which uses near real-time emergency department and ambulance data, was useful for monitoring the outbreak. An alert distributed to clinicians reduced unnecessary hospitalization for patients with suspected sepsis.

We identified a novel human parechovirus (HPeV) type (K251176-02) from a neonate with fever. Analysis of the complete genome showed K251176-02 to be a new HPeV genotype. Since K251176-02 could not be neutralized with antibodies against known HPeV serotypes 1-3, it should be classified as a fourth HPeV serotype.

Three case reports expand the clinical spectrum of HPeV-3 infections and highlight the need to characterize this new pathogen., A third serotype of human parechovirus (HPeV) has been recently isolated from stool specimens of a young Japanese child with transient paralysis. We report 3 additional cases of neonatal sepsis caused by HPeV-3 in the fall of 2001 in Canadian infants 7–27 days old. All children were hospitalized with high fever, erythematous rash, and tachypnea for a median of 5 days. The viruses isolated from nasopharyngeal aspirates grew slowly on tertiary monkey kidney cells and were successfully passaged on Vero cells. The predicted amino acid identity of the VP0-VP3-VP1 region of the three viruses was 74.6%–74.8%, 73.4%–73.6%, and 97.0%–97.1% when compared to HPeV-1, -2, and –3 prototype strains, respectively. Although different, our isolates were closely related; amino acid identity was 99.6%–100% for the last 3 proteins.

We conducted a preliminary comparison of the relative sensitivity of a cross-section of published human rhinovirus (HRV)–specific PCR primer pairs, varying the oligonucleotides and annealing temperature. None of the pairs could detect all HRVs in 2 panels of genotyped clinical specimens; >1 PCR is required for accurate description of HRV epidemiology.

To investigate prevalence and clinical characteristics of klassevirus in South Korea, we performed molecular screening in fecal and nasopharyngeal samples from hospitalized children with gastroenteritis. A total of 26 (8.8%) of 294 fecal samples were positive for klassevirus. Klassevirus may be a possible cause of gastroenteritis.

Diagnostic difficulties may have led to underestimation of rhinovirus infections in long-term care facilities. Using surveillance data, we found that rhinovirus caused 59% (174/297) of respiratory outbreaks in these facilities during 6 months in 2009. Disease was sometimes severe. Molecular diagnostic testing can differentiate these outbreaks from other infections such as influenza.

To assess whether infants hospitalized after an apparently life-threatening event had an associated respiratory virus infection, we analyzed nasopharyngeal aspirates from 16 patients. Nine of 11 infants with positive virus results were infected by rhinoviruses. We detected the new genogroup of rhinovirus C in 6 aspirates. This work was supported by grant PI060532 by Fondo de Investigaciones Sanitarias, Institute of Health Carlos III Sí

To the Editor: Human rhinovirus (HRV) is a common etiologic agent of upper respiratory tract infections and is associated with symptoms such as asthma and wheezing. HRV has >100 serotypes, and recently, several groups reported a new HRV group C (HRV-C) in children that is associated with more severe respiratory infections (1–5). We examined the incidence of respiratory viruses in children hospitalized in Kandang Kerbau Women’s and Children’s Hospital, Singapore (6,7). These studies also identified human metapneumovirus and human bocavirus (HBoV) among children in Singapore. We recently performed a retrospective study by using PCR-based testing (8) to identify HRV, in particular HRV-C, in these patients. From October 2005 through March 2007, a total of 500 nasopharyngeal swab specimens from pediatric patients (age range 1 month through 12 years) were collected and tested for HRVs.

To the Editor: The picornaviruses constitute a large, diverse family of positive-sense RNA viruses, which comprise 8 genera: Enterovirus, Aphthovirus, Cardiovirus, Hepatovirus, Parechovirus, Erbovirus, Kobuvirus, and Teschovirus (1). The genus Kobuvirus contains 2 known species: Aichi virus, which was identified in humans in 1989 and was found to be associated with human acute gastroenteritis (2), and Bovine kobuvirus, which was identified in 2003 in apparently healthy cattle (3). In our study, we identified a candidate novel strain of kobuvirus from porcine fecal specimens; this strain is markedly different from Aichi virus and bovine kobuvirus. Using reverse transcription–PCR (RT-PCR) to characterize calicivirus in porcine fecal specimens with a primer pair of p289/VN3T20 designed for a 3-kb fragment of the virus, we observed an unexpected band on agarose gel electrophoresis (4). After purification and sequencing, the 1,185-bp fragment was found to share 73% similarity with the 3D region of bovine kobuvirus. A pair of primers was then designed from this sequence and synthesized (forward: 5′-TGGACGACCAGCTCTTCCTTAAACAC-3′ and reverse: 5′-AGTGCAAGTCTGGGTTGCAGCCAACA-3′; 495 bp) to screen other porcine samples for the virus by PCR. Our samples were 322 fecal specimens collected during 2006–2007 from healthy piglets

To the Editor: The common cold is regarded as a viral disease. In the first years of the 21st century, several new respiratory viruses have been identified, such as human metapneumovirus (hMPV), coronaviruses NL63 and HKU1, and human bocavirus (HBoV). Many studies have addressed the role of these viruses in hospital settings, but few studies have been conducted among outpatients. We examined the etiology of the common cold in young children who were newly symptomatic but had no need of hospital care. We hypothesized that the etiology could be detected in all cases by using modern diagnostics that test for 16 viruses in outpatients. Between February 1996 and April 1998, we collected nasopharyngeal aspirate samples in an outpatient setting from 194 Finnish children having newly onset (1 respiratory viruses in 92% of the children who had a common cold. As expected, rhinovirus was the leading cause of the common cold in these children. The role of picornaviruses was also emphasized by the abundance of enteroviruses. Enterovirus has gained attention mainly in severe infections, e.g., meningoencephalitis, and is rarely included in diagnostics for respiratory infections. However, PCR has shown that enterovirus commonly causes upper and lower respiratory infections that may be complicated by AOM or expiratory wheezing (4,7). Thus, enterovirus should be included in the diagnostic panels of respiratory infections. HBoV was the second most prevalent virus in our study population. Since its discovery in 2005, HBoV positivity has been reported in 3%–19% of different study populations (8). Its pathogenic role has been questioned because most HBoV cases are co-infections with other viruses (8), and 81% of those testing positive for HBoV in our study had co-infections. However, adenovirus and enterovirus reached similar co-infection frequencies, likely because of prolonged postinfection viral shedding of these agents. HBoV–specific immunoglobulin (Ig) M and IgG antibody responses were recently reported in children with wheezing, suggesting that HBoV induces a systemic infection and is probably a true causative agent of lower respiratory tract disease (9). Our study indicates that HBoV may also be a common cause of common cold in young children. However, we found hMPV, coronaviruses NL63 and HKU1, and influenza C virus in 1%–4% of the children, suggesting that these viruses play a minor role in childhood common cold. Our study may underestimate the role of RSV and hMPV because we excluded children with AOM, which is frequently related to these viruses. Multiple viral findings were common in our study, and 3 children had 4 viruses concomitantly, a logical finding because young children are constantly exposed to respiratory viruses, especially if they attend day care. A recent follow-up study showed that almost all viral findings were related to symptoms, thus supporting the argument that most, if not all, viruses are causative agents (10). A possible causative agent of the common cold can be found in nearly all children who have a cold, and rhinovirus is the leading causative agent. In our study, HBoV was also found frequently, but the recently discovered viruses hMPV and coronaviruses NL63 and HKU1 played a minor role in the common cold of young children.

We appreciate the enthusiasm for our recent publication highlighting the global distribution of a long-unrecognized third clade of rhinoviruses. Robust, sequence-based clock estimates with associated confidence limits indicate that these viruses have been circulating for hundreds of years (1), consistent with the presence of such viruses in historic samples. As isolates from various collections are analyzed in informative regions (e.g., virus protein [VP] 4/2 or VP1), we will undoubtedly find examples in which human rhinoviruses (HRVs) could have been classified as members of the new species HRV-C but were not because the characteristics that define HRV-C were not yet appreciated or because only noncoding sequences had been analyzed. Indeed, we anticipate that waxing interest in HRVs may well lead to the discovery of additional clades. There has been discussion in the field as to whether the novel sequences represent a sublineage HRV-A2 of the classified species HRV-A (2,3), as Mackay et al. had proposed, or whether they should be considered as representatives of a third species of HRV (4,5). The International Committee on Taxonomy of Viruses (ICTV) is charged with the recognition and naming of taxonomic entities. Thus, we provisionally designated our sequences as a novel clade distinct from HRV-A and HRV-B (4) and submitted a proposal to ICTV with data supporting the recognition of HRV-C as a third species of rhinovirus. The proposal was recently approved by the ICTV Study Group on Picornaviruses (Europic May 2008 meeting in Sitges, Spain). Irrespective of taxonomic discourse, we agree with Mackay and colleagues that molecular analyses of as-yet-uncultured HRVs are fascinating and have potential to reveal unexpected insights into the role of HRVs in disease.

To the Editor: Human rhinoviruses (HRVs), members of the family Picornaviridae, were first isolated in 1956 (1); to date, >100 serotypes have been identified on the basis of nucleotide sequence homologies. HRVs were previously divided into 2 genetic groups, HRV-A (n = 75) and HRV-B (n = 25). Recently, a putative new and distinct rhinovirus group, HRV-C, has been reportedly found in some patients with respiratory tract infections (RTIs) (2–8). To extend these initial findings and assess the pathogenicity of HRV-C, we investigated its prevalence as well as its clinical and molecular features in children with lower acute RTIs in Beijing, People’s Republic of China. From July through December 2007, nasopharyngeal aspirates were collected from 258 children (167 boys and 91 girls) who had lower acute RTIs at the time of their admission to Beijing Children’s Hospital. The children were 1 month to 15 years of age (mean age 37 months, median age 10 months). Nucleic acids were extracted from clinical samples by using the NucliSens easyMAG platform (bioMerieux SA, Marcy L’Etoile, France). Each specimen was tested for the presence of common respiratory viruses: human parainfluenza viruses 1–4, influenza viruses, respiratory syncytial virus, enteroviruses, human coronaviruses (229E, NL63, HKU1, and OC43), metapneumovirus, adenoviruses, and bocaviruses. To study the prevalence of HRV-C, we designed a specific reverse transcription–PCR (RT-PCR) that generated a 330-bp PCR product encompassing a portion of the 5′-untranslated region, the full virus capsid protein (VP) 4 gene, and a portion of the VP2 gene of the HRV-C genome. (All primer sequences and protocols of these assays are available from J.W. upon request.) This RT-PCR detected HRV-C in 14 patients (12 boys and 2 girls, 1 month to 13 years of age [mean age 19 months, median age 6 months]). In 6 of the 14 patients, HRV-C was the only virus detected, which suggests a direct correlation between HRV-C infection and lower acute RTIs. In the remaining 8 patients, other respiratory viruses were also detected. Respiratory syncytial virus, the most important cause of lower acute RTIs in children, was codetected in 7 of the HRV-C–positive patients, and human parainfluenza virus 3 was codetected in the other patient. Human coronavirus NL63 was codetected with respiratory syncytial virus in 1 HRV-C–positive patient. HRV-C infection may be seasonal. This virus was detected during only 3 of the 6 months in which specimens were collected. Specifically, HRV-C was detected in samples collected in October (7/50), November (5/96), and December (2/8) but not in those collected in July (0/37), August (0/42), or September (0/25). In contrast, HRV-A and HRV-B were detected in each month (data not shown). Indeed, from July through December, HRV-A and HRV-B viruses were detected in 34 and 12 patients, respectively. Notably, in October 2007, HRV-C was detected in 7 patients, while HRV-A and HRV–B were detected in 5 and 2 patients, respectively, which suggests that the cluster of cases of HRV infections during this month was caused mainly by HRV-C. The 14 HRV-C–positive patients had a variety of other diseases including pneumonia (6/14), bronchopneumonia (4/14), and peribronchiolitis (3/14). The most common clinical findings were cough (14/14), fever (9/14), and abnormal breath sounds on auscultation (11/14). Radiographic results were available for 10 of the 14 HRV-C–positive patients, all of whom had increased lung markings or patchy shadows. Although 3 of the 14 patients required admission to the pediatric intensive care ward, their clinical outcomes were favorable. Phylogenetic analysis showed that the 14 sequences obtained during this study (GenBank accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"EU687515-EU687528","start_term":"EU687515","end_term":"EU687528","start_term_id":"188988671","end_term_id":"188988697"}}EU687515-EU687528) together with previously reported sequences formed a novel group of rhinoviruses (Figure). Six sequences (BCH221, BCH264, BCH200, BCH341, BCH217, and BCH250) displayed high similarity to HRV C025 ({"type":"entrez-nucleotide","attrs":{"text":"EF582386","term_id":"156254958","term_text":"EF582386"}}EF582386) from Hong Kong (4); 2 sequences (BCH249 and BCH343) displayed high similarity to NAT083 ({"type":"entrez-nucleotide","attrs":{"text":"EF077264","term_id":"146269402","term_text":"EF077264"}}EF077264) from the United States (3) and Picornaviridae strain 06-646 ({"type":"entrez-nucleotide","attrs":{"text":"EU081811","term_id":"158538951","term_text":"EU081811"}}EU081811) from Germany (5). BCH242 was similar to Picornaviridae strain tu403 ({"type":"entrez-nucleotide","attrs":{"text":"EU081795","term_id":"158538919","term_text":"EU081795"}}EU081795) from Germany (5). The other 5 strains were homologous to strains from Australia ({"type":"entrez-nucleotide-range","attrs":{"text":"EU155152-EU155154","start_term":"EU155152","end_term":"EU155154","start_term_id":"157837436","end_term_id":"157837440"}}EU155152-EU155154, {"type":"entrez-nucleotide","attrs":{"text":"EU155158","term_id":"157837448","term_text":"EU155158"}}EU155158) (2,7). These findings suggest that, as in other countries (5), the HRV-C strains circulating in China are diverse. Although HRV-C strains belonging to different gene clusters cocirculate, some genetically close strains dominated during certain periods, e.g., the C025-like strain (6/14) was dominant during the study period. A similar distribution pattern is observed in epidemics of HRV-A and HRV-B (9,10). Figure Phylogenetic analysis of the viruses detected in this study based on the nucleotide sequences of the virus capsid protein (VP)4/VP2 region. Using the VP4/VP2 nucleotide sequence (258 nt), we performed neighbor-joining analysis by applying the Kimura 2-parameter ... In conclusion, HRV-C strains were detected in hospitalized children with lower acute RTIs in Beijing. Co-infections were common and complex, which indicates that the role of HRV-C in patients with multiple infections should be further investigated. Our findings provide additional evidence that HRV-C is spreading globally (8) and suggest that HRV-C infections should be considered a serious public health concern.

During an outbreak of severe acute respiratory infections in 2 orphanages, Vietnam, 7/12 hospitalized children died. All hospitalized children and 26/43 children from outbreak orphanages tested positive for rhinovirus versus 9/40 control children (p = 0.0005). Outbreak rhinoviruses formed a distinct genetic cluster. Human rhinovirus is an underappreciated cause of severe pneumonia in vulnerable groups.

This virus typically causes illness in young children but was found to be associated with illness in adults., Human parechovirus has rarely been shown to cause clinical disease in adults. During June–August 2008, a total of 22 adults sought treatment at Yonezawa City Hospital in Yamagata, Japan, for muscle pain and weakness of all limbs; most also had fever and sore throat. All patients received a clinical diagnosis of epidemic myalgia; clinical laboratory findings suggested an acute inflammatory process. Laboratory confirmation of infection with human parechovirus type 3 (HPeV3) was made for 14 patients; we isolated HPeV3 from 7 patients, detected HPeV3 genome in 11, and observed serologic confirmation of infection in 11. Although HPeV3 is typically associated with disease in young children, our results suggest that this outbreak of myalgia among adults was associated with HPeV3 infection. Clinical consideration should be given to HPeV3 not only in young children but also in adults when an outbreak occurs in the community.

To determine possible cosavirus association with clinical disease, we used real-time reverse transcription PCR to test children and HIV-positive adults in Brazil with and without gastroenteritis. Thirteen (3.6%) of 359 children with gastroenteritis tested positive, as did 69 (33.8%) of 204 controls. Low prevalence, frequent viral co-infections, and low fecal cosavirus RNA concentrations argue against human pathogenicity.

Cardioviruses cause myocarditis and encephalomyelitis in rodents; human cardioviruses have not been ascribed to any disease. We screened 6,854 cerebrospinal fluid and 10 myocardium specimens from children and adults. A genotype 2 cardiovirus was detected from a child who died of sudden infant death syndrome, and 2 untypeable cardioviruses were detected from 2 children with meningitis.

To learn more about salivirus/klassevirus, we tested feces of children with diarrhea in China during 2008–2009. We isolated the virus from 9/216 diarrhea samples and 0/96 control samples. The nearly full polyprotein of 1 isolate, SH1, showed 95% identity with a salivirus from Nigeria, indicating widespread distribution and association with diarrhea.

Porcine kobuvirus was first identified in early 2007 in Hungary. Originally thought to be confined to the intestine, almost 2 years later the virus was found in the blood of clinically healthy pigs on the same farm. Porcine kobuvirus may be widely distributed on pig farms worldwide.

Of 362 fecal samples collected from children with acute gastroenteritis in Sri Lanka during 2005–2006, 30 (8.3%) were positive for human parechovirus (HPeV) by reverse transcription–PCR. A novel HPeV, designated as HPeV10, was identified in 2 samples by sequence analysis of the viral protein 1 gene of the detected HPeVs.

To the Editor: Picornaviruses (family Picornaviridae) are small, nonenveloped viruses with single-stranded, positive-sense genomic RNA, currently divided into 8 genera: Enterovirus, Aphthovirus, Cardiovirus, Hepatovirus, Parechovirus, Erbovirus, Teschovirus, and Kobuvirus (1). To date, the genus Kobuvirus consists of 2 species, Aichi virus and Bovine kobuvirus, each possessing 1 serotype. Aichi virus (strain A846/88) was first isolated from a stool sample obtained from a person with acute gastroenteritis in 1991 (2). Bovine kobuvirus (strain U-1) was detected in bovine sera and in feces samples from clinically healthy cattle in 2003 (3). Human and bovine kobuviruses were first isolated in Japan. Recently, kobuviruses have also been detected in humans in other countries in Asia (4), Europe (5,6), and South America (5) and in calves with diarrhea in Thailand (7). The Aichi virus and bovine kobuvirus genomes are approximately 8.2–8.3 kb, respectively, and both have a typical picornavirus genome organization, including the L protein following structural (VP0, VP3, and VP1) and nonstructural (2A–2C and 3A–3D) regions (1,3). Genetic identity between Aichi and U-1 viruses ranges from 47.7% (3′ untranslated region) through 70.8% (3D region) (3). In this study, we report a new candidate species of kobuvirus. Porcine kobuvirus was serendipitously detected in fecal samples from domestic pigs in Hungary. Fecal samples were collected in February 2007 from 15 healthy piglets (Sus scrofa domestica) 2 species (1). Our study reports detection of kobuvirus in domestic pigs. Serendipitously, the generic calicivirus primers p289 and p290, designed for a calicivirus RNA-dependent RNA polymerase region, amplified a kobuvirus 3C/3D region when specimens were tested for porcine caliciviruses by RT-PCR. Comparison of the primers p289 and p290 and the S-1-HUN sequence showed that there are 12- and 11-bp homologous regions between the kobuvirus and the 3′ end of the primer sequences, respectively. Reverse primer p289 designed for calicivirus (norovirus and sapovirus) conserved amino acid 3D motif YGDD, which is also present in kobuviruses. All apparently healthy animals

To the Editor: A new species of kobuvirus, named U-1 strain, was first recognized in 2003 as a cytopathic contaminant in a culture medium of HeLa cells that had been used for >30 years in the laboratory (1). The RNA genome of the U-1 strain comprises 8,374 nucleotides; the genome organization is analogous to that of picornaviruses. Morphologically, the U-1 strain resembles the Aichi virus, but genetically it is distinct (1). Therefore, the U-1 strain is classified as a new species of genus Kobuvirus in the family Picornaviridae, and it is called bovine kobuvirus (1). To date, the genus Kobuvirus consists of 2 species, Aichi virus and bovine kobuvirus (2). The Aichi virus is associated with acute gastroenteritis in humans (3–5); bovine kobuvirus infection has been detected only in cattle (1). Only 1 report has described the discovery and epidemiologic features of bovine kobuvirus (1). Of serum samples from 72 healthy cattle, 43 (59.7%) were positive for neutralizing antibody against bovine kobuvirus U-1 standard strain at a titer of >16. In addition, 12 (16.7%) of 72 stool samples collected from the cattle were positive for the bovine kobuvirus genome by reverse transcription–PCR (RT-PCR) (1). This finding suggested that bovine kobuvirus is common and that the virus particles could be detected in the stool samples of infected cattle. We therefore conducted an epidemiologic survey of bovine kobuvirus and report detection of this virus in stool samples from calves with diarrhea during 2001–2004 in Chiang Mai Province, Thailand. From November 2001 to July 2004, a total of 72 fecal specimens were collected. The age of the calves ranged from 7 to 49 days. The presence of bovine kobuvirus in fecal specimens was detected by using RT-PCR with a protocol modified from the method described by Yamashita et al. (1). All the bovine kobuvirus strains detected in our study were analyzed further by direct sequencing of their PCR amplicons with the BigDye Terminator Cycle Sequencing Kit (Applied Biosystems, Foster City, CA, USA) on an automated sequencer (ABI 3100, Applied Biosystems). The nucleotide sequences of these portions were compared with those of reference strains available in the GenBank database by using BLAST (6). Phylogenetic and molecular evolutionary analyses were conducted by using MEGA version 3.1 (7). The nucleotide sequences of bovine kobuvirus strains described in this study were deposited in GenBank under accession nos. {"type":"entrez-nucleotide-range","attrs":{"text":"EF659450-EF659455","start_term":"EF659450","end_term":"EF659455","start_term_id":"151299687","end_term_id":"151299697"}}EF659450-EF659455. The bovine kobuvirus was detected by the RT-PCR screening method in 6 (8.3%) of the 72 fecal specimens collected. The partial 3D regions of all 6 bovine kobuviruses exhibited highly conserved sequences of 99.3%–100% nucleotide and 100% amino acid identities to each other. In addition, by searching for the closest sequences in the databank, we found that these sequences were closely related to all 13 bovine kobuvirus reference strains available in the GenBank database (U-1, K-35, K-36, K-49, K-38, N-5, K-3, K-4, K-6, K-60, N-2, K-44, and K-55); nucleotide and amino acid sequence identities ranged from 91.0% to 95.0% and 97.4% to 99.4%, respectively. Phylogenetic analysis of partial 3D nucleotide sequences of our bovine kobuvirus strains, together with those of all published bovine kobuvirus reference strains available in the GenBank database, is shown in the Figure. The Aichi virus standard strain was included in the tree as an outlier virus. The phylogenetic tree confirmed that all 6 virus strains belonged to bovine kobuvirus and formed a tight cluster in a monophyletic branch with other published bovine kobuvirus reference strains, but they were distantly related to the Aichi virus standard strain. Figure Phylogenetic analysis of the partial nucleotide sequence encoding the 3D region of bovine kobuviruses isolated in this study and other reference strains recognized to date. The tree was generated on the basis of the neighbor-joining method using the MEGA ... Detection and characterization of bovine kobuvirus strains from different geographic areas are important for understanding the worldwide distribution, heterogeneity, and association of bovine kobuvirus with enteric disease in cattle. Our findings indicate the role of kobuviruses in diarrheal disease in cattle and provide additional information on their relationship to bovine kobuviruses reported previously.

These infections are associated with a variety of clinical syndromes, in part related to specific serotype., A new reverse transcription–polymerase chain reaction assay was developed for identification of 28 Canadian human parechovirus (HPeV) isolates, including 20 HPeV-1, 3 HPeV-2, and 5 HPeV-3, recovered from 1985 to 2004. All HPeV-1 isolates but 1 were genetically distinct from the Harris reference strain. One HPeV-2 isolate was related to the Williamson strain; the other 2 were related to the Connecticut strain. HPeV-3 isolates clustered together. Seventy-five percent of isolates were recovered during the typical enterovirus season. All patients but 1 were children with a mean age of 14.6 months, 6.3 months, and 0.7 months for HPeV-1, HPeV-2, and HPeV-3 patients, respectively. All HPeV-2– and HPeV-3–infected children were hospitalized with a diagnosis of viremia or sepsis. HPeV-1–infected children had bronchiolitis diagnosed in 50% of the cases, with few cases of pneumonia and enteritis. Two infected patients (1 child with leukemia and a 78-year-old woman) died of septic shock and severe pneumonia, respectively.

Nosocomial viral respiratory infections cause considerable illness and death on pediatric wards. Common causes of these infections include respiratory syncytial virus and influenza. Although primarily a community pathogen, rhinovirus also occasionally results in hospitalization and serious sequelae. This article reviews effective infection control interventions for these three pathogens, as well as ongoing controversies.

To the Editor: Picornaviruses (family Picornaviridae) are small, nonenveloped viruses with single-stranded, positive-sense genomic RNA. They are divided into 12 genera: Enterovirus, Aphthovirus, Cardiovirus, Hepatovirus, Parechovirus, Erbovirus, Teschovirus, Sapelovirus, Senecavirus, Tremovirus, Avihepatovirus, and Kobuvirus. The genus Kobuvirus consists of 2 officially recognized species, Aichi virus (1) and Bovine kobuvirus (2), and 1 candidate species, porcine kobuvirus (3). The kobuvirus genome is ≈8.2–8.4 kb long and has the typical picornavirus genome organization of leader (L) protein following the structural (viral protein [VP] 0, VP3, and VP1) and nonstructural (2A–2C and 3A–3D) regions (2,4). The genetic identity on the coding region between Aichi (strain A846/88), bovine (U-1), and porcine (S-1-HUN) viruses is between 35% (L protein) and 74% (3D region) (2,4). Aichi virus and bovine kobuvirus were first detected in fecal samples from humans and cattle in Japan, in 1991 and 2003, respectively (1,2). Porcine kobuvirus was identified from domestic pigs in Hungary in 2008 (3,4). Recent studies demonstrated that Aichi virus circulates in Asia (5), Europe (6,7) including Hungary (4), South America (6), and North Africa (8) and can cause gastroenteritis in humans. In addition, bovine and porcine kobuviruses are detected among these farm animals in Europe (4) and Asia (2,9). These data indicate that kobuviruses are widely distributed geographically and raise the possibility of additional animal host species. We detected kobuvirus in sheep. On March 17, 2009, a total of 8 fecal samples were collected from young, healthy, domestic sheep (Ovis aries)

To the Editor: Picornaviruses (family Picornaviridae) are small, nonenveloped viruses with single-stranded, positive-sense genomic RNA. Picornaviruses are currently divided into 8 genera: Enterovirus, Aphthovirus, Cardiovirus, Hepatovirus, Parechovirus, Erbovirus, Teschovirus, and Kobuvirus (1). To date, the genus Kobuvirus consists of 2 officially recognized species, Aichi virus and Bovine kobuvirus, and 1 porcine kobuvirus as a candidate species (2–4). Aichi virus (strain A846/88) was first isolated in 1991 from feces of a person with acute gastroenteritis (2). Bovine kobuvirus (strain U-1) was detected in 2003 in bovine serum and fecal samples from clinically healthy cattle (3); in 2008, it was isolated from cattle with diarrhea (5). Aichi virus and bovine kobuvirus were first isolated in Japan. Porcine kobuvirus (strain S-1-HUN) was recently identified from domestic pigs in Hungary (4). Aichi viruses have been also detected in other countries in Asia (6), Europe (7,8), South America (7), and northern Africa (9). Bovine kobuvirus, however, has not been detected outside Asia (Japan and Thailand) (3,5).