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Porcine rotavirus is one of the most important pathogens causing diarrhea in newborn piglets, and the genome of this virus contains 11 double-stranded RNA segments, which are easy to be recombined among strains to produce new strains with different antigenic properties. The reverse genetics system is an informative tool for studying virus biology. Recently, adaptable plasmid-based reverse genetics systems were developed for the porcine rotavirus OSU strain; however, such systems have not been developed for epidemic porcine rotavirus genotypes in China. In this study, we successfully established a reverse genetic system based on an epidemic strain of porcine rotavirus JXAY01 isolated in recent years, which was characterized by a specific genotype constellation: G5-P[23]-I12-R1-C1-M1-A8-N1-T7-E1-H1. 11 gene segments of porcine rotavirus JXAY01 were cloned into plasmid vectors similar to the SA11 system. JXAY01 genome segment plasmids were co-transfected with 10 complementary SA11 genome plasmids, and 11 monoreassortant strains were successfully rescued. Viral replication analyses of the parental SA11 strain and the monoreassortant strains showed that the structural protein replacement monoreassortants had reduced cell proliferation compared with the parental SA11 and non-structural protein replacement strains. The recombinant rJXAY01 strain could be successfully rescued using 11 pRG-JXAY01 plasmids. Whole genome sequencing showed 12 amino acid differences between the isolate JXAY01 and the recombinant rJXAY01, but there was no significant difference in their in vitro replication ability. This study reports the reverse genetic system, which lays the foundation for further understanding of porcine rotavirus molecular biology and novel vaccine development. [ABSTRACT FROM AUTHOR]

Rotavirus (RV) is a leading pathogen causing diarrhea in children. In this study, a total of 51 fecal samples from children with RV enteritis, 29 post-treatment fecal samples, and 38 fecal samples from age-matched healthy controls were collected. Microbial DNA was isolated from the samples followed by high throughput Illumina sequencing targeting 16 S rRNA gene. Compared to the healthy group, the RV-infected group exhibited reduced microbial diversity. Both groups shared Firmicutes as the dominant phylum. Additionally, the abundance of Proteobacteria increased significantly in the RV-infected group. At the genus level, among the top 50 most abundant genera, 34 showed significant differences, with these differential genera correlating with certain clinical indicators such as dehydration levels and C-reactive protein (CRP). Notably, there were no significant differences in the microbiota before and after treatment in RV-infected children. Only 8.82% (3/34) of the differential genera in the post-treatment group showed a recovery trend towards the healthy state. This study enhances the understanding of how RV infection alters the gut microbiota structure in children and provides a scientific basis for improving clinical diagnosis and treatment strategies. [ABSTRACT FROM AUTHOR]

Viruses engage in a variety of processes to subvert host defenses and create an environment amenable to replication. Here, using rotavirus as a prototype, we show that calcium conductance out of the endoplasmic reticulum by the virus encoded ion channel, NSP4, induces intercellular calcium waves that extend beyond the infected cell and contribute to pathogenesis. Viruses that lack the ability to induce this signaling show diminished viral shedding and attenuated disease in a mouse model of rotavirus diarrhea. This implicates nonstructural protein 4 (NSP4) as a virulence factor and provides mechanistic insight into its mode of action. Critically, this signaling induces a transcriptional signature characteristic of interferon-independent innate immune activation, which is not observed in response to a mutant NSP4 that does not conduct calcium. This implicates calcium dysregulation as a means of pathogen recognition, a theme broadly applicable to calcium-altering pathogens beyond rotavirus. [ABSTRACT FROM AUTHOR]

Rotavirus A (RVA) is the primary enteric pathogen of humans and many other species. However, RVA interspecies transmission remains poorly understood. In this study, we conducted a comprehensive screening and genotyping analysis of RVA in 1706 wild animal samples collected from various regions within Yunnan Province, China. A total of 24 samples, originating from wild boars, rodents, bats, and birds tested positive for RVA. Next generation sequencing and phylogenetic analyses revealed a high degree of genetic diversity and reassortment, particularly for VP4 and VP7. Strains isolated from wild boars and rodents exhibited gene segments with high similarity to those found in humans and other mammalian RVA strains, indicating that RVA may undergo interspecies transmission and reassortment, resulting in novel strains with potential risks for human infection. This study provides critical data for understanding the transmission mechanisms and the RVA host range, and highlights the pivotal role of wildlife in viral evolution and dissemination. These findings have significant implications for public health policies and emphasize the need for enhanced surveillance to prevent interspecies RVA transmission. [ABSTRACT FROM AUTHOR]

Background: Viral gastroenteritis is a significant global health concern. An effective, rapid, and easy-to-use diagnostic tool is essential for screening causative viruses. Methods: Forty-eight samples, known to be infected with one of the following viruses: norovirus, group A rotavirus, astrovirus, adenovirus, and sapovirus determined by reverse transcription-PCR and nucleotide sequencing, were evaluated by the Fast Track Diagnostics (FTD) viral gastroenteritis assay. Results: The assay demonstrated 100% specificity for all viruses and matched the RT-PCR sensitivity for norovirus GI, classic human astrovirus, adenovirus, and sapovirus. It identified norovirus GII and rotavirus with 87.5% and 85.7% sensitivity, respectively. However, its sensitivity for detecting novel human astrovirus MLB and VA was lower, at 35%. Conclusions: The FTD viral gastroenteritis assay can effectively screen simultaneously for norovirus GI, GII, group A rotavirus, adenovirus, and sapovirus in clinical settings. The study also suggests that improved detection methods are necessary for novel astrovirus strains. [ABSTRACT FROM AUTHOR]

There is currently limited evidence regarding how the rotavirus vaccine dosing schedule might be adjusted to improve vaccine performance. We quantified the impact of the previously implemented 6/10-week Rotarix vaccine (RV1) in Ghana to the model-predicted impact for other vaccine dosing schedules across three hospitalsand the entire country. Compared to no vaccination, the model-estimated median percentage reductions in rotavirus ranged from 28 to 85% and 12 to 71% among children <1 and <5 years old, respectively. The median predicted reductions in rotavirus for the whole country ranged from 57 to 66% among infants <1 year and 35 to 45% among children <5 years old. The 1/6/10- and 6/10/14-week schedules provided the best and comparable reductions in rotavirus compared to the original 6/10-week schedule. A third dose could prevent an additional 9 to 14% of deaths. An additional dose of RV1 might be an effective strategy to improve rotavirus vaccine impact, particularly in settings with low vaccine effectiveness. [ABSTRACT FROM AUTHOR]

Introduction: Rotavirus is an important cause of severe diarrhoea in children under five years of age, accounting for approximately 78,000 deaths annually in this age group. The magnitude of acute gastroenteritis caused by rotavirus is often underestimated due to the lack of investigations into these cases. Although rotavirus disease is less common in adults, it can occur, possibly because of contact with children who have rotavirus gastroenteritis. Aim: To assess the current trend of the disease in both children as well as adults presenting with acute diarrhoea. Materials and Methods: This cross-sectional study was conducted over a period of one year (January 2023 to January 2024). A total of 185 stool samples were collected from children under five years of age and adults over 18 years presenting with acute diarrhoea at the outpatient and inpatient departments of Guru Nanak Dev Hospital (GNDH), Amritsar, Punjab, India. Commercially available Enzyme Linked Immunosorbent Assay (ELISA) kits were used to detect the VP6 antigen of Group A rotavirus in the stool samples of the patients. The data collected were analysed using Statistical Package for the Social Sciences (SPSS), and relevant conclusions were drawn. Results: Out of the 185 samples collected, 32 samples (17.3%) tested positive for rotavirus antigen. Vaccinated children experienced milder disease compared to unvaccinated ones. Non breastfed infants showed a higher detection rate of rotavirus, with 20 children (80%) testing positive compared to their breastfed counterparts. The rural-urban distribution indicated a greater burden in rural areas, with 16 children (64%) affected. Conclusion: This study identifies the risk factors, clinical profile, current burden and seasonal variation of rotavirus diarrhoea. It will be helpful in evaluating the impact of the rotavirus vaccine on the severity of the disease. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine rotavirus is a significant diarrhea virus in pig farming, particularly causing fatal dehydration in suckling piglets younger than 7 days, thereby resulting in substantial economic losses to the swine industry. In this study, we have established a rapid detection and analysis system for Group A porcine rotavirus using Recombinase Aided Amplification (RAA) combined with Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology for the first time. The recombinant plasmid PoRV achieved a high-sensitivity response within 30 min at 37 °C with a detection limit as low as 2.43 copies/μL, representing a 10-fold higher sensitivity compared to the qPCR method. Specificity testing results showed that the RAA-CRISPR/Cas12a analysis for porcine rotavirus did not react with other common porcine diarrhea viruses. Furthermore, this method allows visualization of results using blue light, making this accurate and portable detection method hold great potential for rotavirus control in pig farming. Piglet diarrhea poses significant economic losses to the pig industry, posing a worldwide challenge that urgently needs to be addressed in pig breeding practices. Porcine rotavirus (PoRV) is an important viral diarrhea pathogen in piglets, with a high incidence rate and a tendency to cause growth retardation. To enhance the sensitivity and specificity of PoRV detection, we sequenced the NSP3 gene of G5 and G9 genotypes of rotavirus A (RVA), enabling simultaneous detection of the two serotypes. Subsequently, we developed a rapid PoRV detection method using a combination of recombinase-aided amplification (RAA) and CRISPR/Cas12a. In this method, Cas12a binds to RAA amplification products, guided by CRISPR-derived RNA (crRNA), which activates its cleavage activity and releases fluorescence by cutting FAM-BHQ-labeled single-stranded DNA (ssDNA). In the optimized reaction system, the recombinant plasmid PoRV can achieve a highly sensitive reaction within 30 min at 37 °C, with a detection limit as low as 2.43 copies/μL, which is ten times higher in sensitivity compared to the qPCR method. Results from specificity testing indicate that no cross-reactivity was observed between the RAA-CRISPR/Cas12a analysis of PoRV and other viral pathogens, including PoRV G3, PoRV G4, porcine epidemic diarrhea virus (PEDV), porcine epidemic diarrhea (PDCoV), and porcine reproductive and respiratory syndrome virus (PRRSV). In the clinical sample detection using the RAA-CRISPR/Cas12a method and qPCR, Cohen's Kappa value reached as high as 0.952. Furthermore, this approach eliminates the need for large-scale instrumentation, offering a visual result under an ultraviolet lamp through fluorescence signal output. [ABSTRACT FROM AUTHOR]

Using technologies and enforcing national reuse guidelines helps reduce health risks from recycled water used in irrigation. Analysing viral presence in raw and treated sewage is crucial for preventing pandemics. This study used qPCR to evaluate three urban wastewater treatment plants in Ramallah: Al-Tireh MBR, Al-Bireh AS, and Rawabi MBBR. Samples collected in March 2019 were treated and analysed using real-time PCR with specific primers and probes. Results showed Al-Tireh MBR achieved 100% removal of enterovirus, adenovirus, norovirus, and rotavirus. Al-Bireh WWTP achieved 33% removal for adenovirus and rotavirus. Rawabi MBBR achieved varying removal rates: 100% for adenovirus, 33% for norovirus, and 66% for rotavirus. Chlorinated samples showed no direct correlation between faecal coliforms and viral presence. These findings suggest retrofitting Al-Bireh WWTP and Rawabi MBBR with ultrafiltration or MBR units to mitigate viral outbreaks. [ABSTRACT FROM AUTHOR]

Reverse genetics systems for rotaviruses (RV) facilitate the generation of genetically engineered RVs by transfection of 11 plasmids encoding 11 genomic viral RNA segments. In addition to viral genome expression, overexpression of NSP2 and NSP5 has been used to increase the rescue efficiency of recombinant RVs. Here, we showed that the overexpression of nucleotide sequence-modified NSP2 and NSP5 enabled the rapid and efficient production of recombinant RVs. Using improved reverse genetics, we established a reverse genetics system for human and bovine RV clinical isolates, as well as laboratory strains of bovine RV (NCDV and UK) and porcine RV (Gottfried). In addition, we rescued low-replicating recombinant RVs carrying a mutant NSP4 lacking the double-layered particle-binding domain, which was deficient in the efficient production of mature virions. These advancements in reverse genetics enabled the generation of molecular clones of RV clinical isolates and recombinant RVs harboring critical amino acid mutations, offering a versatile platform for investigating RV biology and pathogenesis. [ABSTRACT FROM AUTHOR]

Simple Summary: Acute gastroenteritis (AGE) in cattle is a major economic concern, causing illness and reduced productivity. Bovine Rotavirus A (BoRVA) and bovine coronavirus (BCoV) are two key viruses responsible, leading to diarrhea and other health issues in cattle. While BoRVA mainly affects young calves, BCoV can impact cattle of all ages, causing digestive and respiratory clinical signs. This concise review examines the structure, spread, clinical signs, diagnosis, treatment, and prevention of BoRVA and BCoV, highlighting their potential to spread across species and the importance of effective management practices to control their impact. Acute gastroenteritis (AGE) in cattle significantly impacts the economy due to relatively high morbidity and mortality and decreased production. Its multifactorial nature drives its global persistence, involving enteric viruses, bacteria, protozoa, and environmental factors. Bovine Rotavirus A (BoRVA) and bovine coronavirus (BCoV) are among the most important enteric RNA viruses causing AGE in cattle. These viruses infect intestinal enterocytes, leading to cell damage and consequently to malabsorption and diarrhea. BoRVA primarily affects calves under 14 days old with gastrointestinal clinical signs, while BCoV affects all ages, causing gastrointestinal and respiratory distress. The economic impact of BoRVA and BCoV, along with their interspecies transmission potential, warrants attention. This concise review discusses the molecular structure, epidemiology, pathogenesis, clinical signs, diagnosis, treatment, and preventive measures of BoRVA and BCoV while providing a comparative analysis. By offering practical guidance on managing such viral infections in cattle, these comparative insights may prove valuable for veterinarians in clinical practice. [ABSTRACT FROM AUTHOR]

Inter‐genogroup reassortment of Rotavirus A (RVA) strains has highlighted the spread of unusual RVA strains worldwide. We previously reported the equine‐like G3 RVA as the predominant strain in Indonesia in 2015–2016. However, since July 2017, typical human genotypes G1 and G3 have replaced these strains completely. To understand how dynamic changes in RVA occur in Indonesia, we performed a detailed epidemiological study. A total of 356 stool specimens were collected from hospitalized children in Sidoarjo, Indonesia between 2018 and 2022. Whole‐genome sequencing was performed for all 26 RVA‐positive samples using next‐generation sequencing. Twenty‐four samples were determined to be the unusual RVA G9P[4], while two were G9P[6]. Detailed analysis revealed that seven G9P[4] strains had the typical DS‐1‐like backbone, while the other strains exhibited a double‐reassortant profile (G9‐N1) on the DS‐1‐like backbone. The Bayesian evolutionary analyses suggested that the Indonesian G9P[4] strains share a common ancestor with previously reported G9P[4] strains in the VP7 and VP4 genes. G9P[4] DS‐1‐like strains were identified as the predominant genotype in Indonesia in 2021 for the first time. These results suggest that the G9P[4] strains were generated from the previous G9P[4] strains that had undergone further intra‐reassortments with the other circulating strains. [ABSTRACT FROM AUTHOR]

Porcine astrovirus (PoAstV), porcine sapovirus (PoSaV), porcine norovirus (PoNoV), and porcine rotavirus A (PoRVA) are newly discovered important porcine diarrhea viruses with a wide range of hosts and zoonotic potential, and their co-infections are often found in pig herds. In this study, the specific primers and probes were designed targeting the ORF1 gene of PoAstV, PoSaV, and PoNoV, and the VP6 gene of PoRVA. The recombinant standard plasmids were constructed, the reaction conditions (concentration of primers and probes, annealing temperature, and reaction cycle) were optimized, and the specificity, sensitivity, and reproducibility were analyzed to establish a quadruplex real-time quantitative RT-PCR (RT-qPCR) assay for the detection of these four diarrheal viruses. The results demonstrated that the assay effectively tested PoAstV, PoSaV, PoNoV, and PoRVA without cross-reactivity with other swine viruses, and had limits of detection (LODs) of 138.001, 135.167, 140.732, and 132.199 (copies/reaction) for PoAstV, PoSaV, PoNoV, and PoRVA, respectively, exhibiting high specificity and sensitivity. Additionally, it displayed good reproducibility, with coefficients of variation (CVs) of 0.09–1.24% for intra-assay and 0.08–1.03% for inter-assay. The 1578 clinical fecal samples from 14 cities in Guangxi Province, China, were analyzed via the developed assay. The results indicated that the clinical samples from Guangxi Province exhibited the prevalence of PoAstV (35.93%, 567/1578), PoSaV (8.37%, 132/1578), PoNoV (2.98%, 47/1578), and PoRVA (14.32%, 226/1578), and had a notable incidence of mixed infections of 18.31% (289/1578). Simultaneously, the 1578 clinical samples were analyzed with the previously established assays, and the coincidence rates of these two approaches exceeded 99.43%. This study developed an efficient and precise diagnostic method for the detection and differentiation of PoAstV, PoSaV, PoNoV, and PoRVA, enabling the successful diagnosis of these four diseases. [ABSTRACT FROM AUTHOR]

Copyright of Kocatepe Veterinary Journal / Kocatepe Veteriner Dergisi is the property of Afyon Kocatepe University, Faculty of Veterinary Medicine and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Pangolins have been found to carry severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2)‐related coronaviruses. In light of this discovery, interest has been piqued in viromes of these heavily trafficked wild animals. In this study, we performed viral metagenomic sequencing to explore viromes of both confiscated dead pangolins and captive healthy pangolins. Sequence reads of vertebrate‐associated viruses in Herpesviridae, Retroviridae, Iridoviridae, Reoviridae, Arenaviridae, and Flaviviridae were detected in confiscated dead pangolins. A novel rotavirus (RV) (Reoviridae), showing a high degree of genetic similarity to the RV species D (RVD) that was previously unreported in mammals, was further confirmed by using reverse transcription‐polymerase chain reaction (RT‐PCR) and Sanger sequencing. Three out of 18 samples from the confiscated dead pangolins were positive for genomic sequences of the novel RV. Importantly, sequence alignments and phylogenetic analyses demonstrated that these RV strains genetically belonged to the RVD. Nevertheless, these novel RVD strains were divergent from known RVD strains that have been found only in Avian. They formed a separate genetic cluster. Five serial passages were attempted to isolate the RV, but no live virus was obtained. In addition, fecal samples were collected from healthy pangolins (n = 41) in our institution and screened for RVs by viral metagenomic sequencing and RT‐PCR. In these fecal samples, neither species D nor previously identified species A RVs were detected. This study reported RVDs in pangolin samples for the first time to our knowledge. Identifiability disagreements between wild and captive pangolins highlight the need for further exploration into pangolin viruses to better understand their emergence and transmission potential. [ABSTRACT FROM AUTHOR]

Rotaviruses are well‐recognized pathogens responsible for diarrhea in humans and various animal species, with Rotavirus A the most often detected and most thoroughly described. Rotaviral disease is an important concern in pathology of pigeons as well, as pigeon rotavirus A was proven to play a major role in young pigeon disease (YPD). However, rotaviruses of other groups have been so far understudied in birds. This paper describes the first finding of Rotavirus G in domestic pigeon in Europe, as well as the recovery of three complete genomes of pigeon rotavirus A with Oxford Nanopore Sequencing. Quantification of pigeon rotavirus A genetic material with droplet digital polymerase chain reaction (PCR) in pigeons suffering from diarrhea and in asymptomatic pigeons was also performed in the frameworks of this study and resulted in determination of statistically highly significant differences between the groups in both detection rate and shedding of the virus. Phylogenetic analysis revealed the close relationship of acquired strains with those originating from pigeons from Europe, North America, Asia, and Australia, indicating a broad geographical spread of pigeon rotaviruses. Results of our research shed more light on occurrence and diversity of Rotavirus species in pigeons. [ABSTRACT FROM AUTHOR]

The causes of diarrhea after 10 years of rotavirus vaccination in Rwanda were investigated with real-time polymerase chain reaction in 496 children with diarrhea and 298 without. Rotavirus was detected in 11% of children with diarrhea (odds ratio, 2.48; P =.002). Comparison of population attributable fractions (PAFs) shows that Shigella (PAF, 11%) and enterotoxigenic Escherichia coli producing labile toxin (PAF, 12%) have replaced rotavirus as the main causative agents. The PAF for rotavirus had declined from 41% prevaccination to 6.5% postvaccination, indicating that rotavirus has become one among several similarly important causes of childhood diarrhea in Rwanda. A rotavirus genotype shift to G3P[8] points at the importance of continued genotype surveillance. [ABSTRACT FROM AUTHOR]

Background Quantitative molecular assays are increasingly used for detection of enteric viruses. Methods We compared the clinical severity using the modified Vesikari score (mVS) of enteric viruses detected by conventional assays (enzyme immunoassays [EIAs] for rotavirus and adenovirus 40/41 and conventional polymerase chain reaction for astrovirus, sapovirus, and norovirus) and a quantitative molecular assay (TaqMan Array Card [TAC]) among children aged 0–59 months in the Global Enteric Multicenter Study. For rotavirus and adenovirus 40/41, we compared severity between EIA-positive and TAC-positive cases assigned etiologies using different cycle threshold (Ct) cutoffs. Results Using conventional assays, the median mVS (interquartile range) was 10 (8–11) for rotavirus, 9 (7–11) for adenovirus 40/41, 8 (6–10) for astrovirus, sapovirus, and norovirus GII, and 7 (6–9) for norovirus GI. Compared with rotavirus EIA-positive cases, the median mVS was 2 and 3 points lower for EIA-negative/TAC-positive cases with Ct <32.6 or Ct ≥32.6 and <35, respectively (P <.001). Adenovirus 40/41 EIA-positive and EIA-negative/TAC-positive cases were similar, regardless of Ct cutoff. Conclusions Quantitative molecular assays compared with conventional assays, such as EIA, may influence the severity of identified cases, especially for rotavirus. Cutoffs to assign etiology for quantitative assays should be considered in the design and interpretation of enteric virus studies. [ABSTRACT FROM AUTHOR]

Background: Infection is the cause of diarrhoea, and rotaviruses and adenoviruses are important pathogens in children. Methods: A retrospective study was conducted on 144,067 children with diarrhoea between 2014 and 2023 in China. We used the colloidal gold method to detect intestinal adenovirus and rotavirus A antigens in faeces. The epidemiological characteristics of these viruses and the impact of meteorological factors on them were analysed before and after coronavirus disease 2019 (COVID-19) pandemic. Results: During this decade, the positive rate of adenovirus infection was 6.41%, while the positive rate of rotavirus A infection was 11.81%, higher than that of adenovirus infection. The positive rate of adenovirus and rotavirus A coinfection was 1.92%. The positive rates of adenovirus, rotavirus A and coinfection showed a fluctuating trend, and suddenly decreased in 2020. There was an apparent decrease of positive rate of rotavirus A, with a decrease of 57.27%, during 2020–2023. Surprisingly, the positive rate of adenovirus infection exceeded that of rotavirus A infection in 2021 and 2023. During the COVID-19 pandemic, the proportion of female patients and children over two years of age infected with adenovirus or rotavirus A increased, while the proportion of cases in winter decreased. In addition, we found that the positive rate of rotavirus A infection was related to average temperature and sunshine, and the positive rate of adenovirus and rotavirus A coinfection was only related to sunshine. However, these correlations disappeared during the COVID-19 pandemic. Conclusions: This study revealed the recent prevalence of adenovirus and rotavirus A infections in children with diarrhoea in south-central China and provided a theoretical basis for the prevention and control of viral diarrhoea. [ABSTRACT FROM AUTHOR]

Background: Rotaviruses are the major etiological agents of gastroenteritis and diarrheal outbreaks in plenty of mammalian species. The genus Rotavirus is highly diverse and currently comprises nine genetically distinct species, and four of them (A, B, C, and H) are common for humans and pigs. There is a strong necessity to comprehend phylogenetic relationships among rotaviruses from different host species to assess interspecies transmission, specifically between humans and livestock. To reveal the genetic origin of rotaviruses from Russian pig farms, nanopore-based metagenomic sequencing was performed on the PCR-positive specimens. Methods: Samples were selected among the cases submitted to routine diagnostic or monitoring studies to the Laboratory of Biochemistry and Molecular Biology of "Federal Scientific Center VIEV" (Moscow, Russia). The selected positive samples were genotyped using nanopore sequencing method. Results: Five porcine RVA isolates were completely sequenced, and genotype analysis revealed various porcine G/P genogroups: G2, G3, G4, G5, G11 and P[6], P[7], P[13], P[23], P[27] with a typical backbone constellation I5-R1-C1-M1-A8-N1-T1/7-E1-H1. The RVB isolate was detected in combination with RVA in a rectal swab from a diseased pig in Krasnoyarsk Krai. It was characterized by the following genogroups: G15-P[X]-I11-R4-C4-M4-A8-N10-T4-E4-H7. The first complete porcine RVC genome from Russia was obtained with genomic constellation G6-P[5]-I14-R1-C1-M1-A7-N9-T6-E1-H1, and the phylogenetic analysis revealed putative novel genotype group for the VP6 gene-I14. Additionally, the first porcine kobuvirus isolate from Russia was phylogenetically characterized. Conclusions: The applied nanopore sequencing method successfully genotyped the RV isolates and additionally revealed co-circulated species. The study demonstrates high genetic variability of Russian RVA isolates in VP4/VP7 genes and phylogenetically describes local RVB and RVC. Complete characterization of genomic segments is a crucial methodology in tracing the rotavirus's evolution and evaluating interspecies transmissions. [ABSTRACT FROM AUTHOR]

Rotaviruses pose a significant threat to young children. To identify novel pro- and anti-rotavirus host factors, we performed genome-wide CRISPR/Cas9 screens using rhesus rotavirus and African green monkey cells. Genetic deletion of either SERPINB1 or TMEM236, the top two antiviral factors, in MA104 cells increased virus titers in a rotavirus strain independent manner. Using this information, we optimized the existing rotavirus reverse genetics systems by combining SERPINB1 knockout MA104 cells with a C3P3-G3 helper plasmid. We improved the recovery efficiency and rescued several low-titer rotavirus reporter and mutant strains that prove difficult to rescue otherwise. Furthermore, we demonstrate that TMEM236 knockout in Vero cells supported higher yields of two live-attenuated rotavirus vaccine strains than the parental cell line and represents a more robust vaccine-producing cell substrate. Collectively, we developed a third-generation optimized rotavirus reverse genetics system and generated gene-edited Vero cells as a new substrate for improving rotavirus vaccine production. [ABSTRACT FROM AUTHOR]

Rotavirus vaccines carry a small risk of intussusception mainly 1–7 days after vaccination in the United States of America, Europe, Australia, and Latin America where the background rate of intussusception is relatively low. Such risks are undetectable in Africa and India where the background rate is the lowest. Because few studies were carried out in high-background-rate countries such as Japan, we examined how intussusception occurred in infants living in Akita prefecture, Japan, while the vaccines were sold in the private market. Between 2011 and 2018, an estimated 21,677 infants (46%) were vaccinated and 54% were not. Through a retrospective survey of medical records in 18 hospitals in the prefecture, we identified 58 infants, 28 of whom were vaccinated and 30 of whom were unvaccinated, as having intussusception that met level 1 of the Brighton criteria. Thus, the intussusception rate was 123 per 100,000 infant-years (95% confidence interval [CI]: 94–160). Despite the high rate, none developed intussusception 1–7 days after the first dose of either the monovalent human rotavirus vaccine (GSK) or the pentavalent human–bovine reassortant vaccine (MSD). The incidence rate ratio of vaccinated to unvaccinated infants between 42 and 245 days of life was estimated at 0.96 (95%CI: 0.43–2.1; p = 0.92). Given that over 95% of infants received the first dose before 15 weeks of age, the risk of intussusception associated with the rotavirus vaccines in high-incidence-rate countries can be reduced to a minimum by adhering to the recommended schedule at 2, 3, and 4 months of age. [ABSTRACT FROM AUTHOR]

Rotavirus species A (RVA) is a major cause of acute viral gastroenteritis in young humans and diverse animal species. The study of the genetic characteristics of RVAs that infect rabbits (Oryctolagus cuniculus) (lapine strain [LRV]) has been limited, and, to date, the most common and epidemiologically important combinations of G/P genotypes in rabbits have been reported to be G3 P[14] and G3 P[22]. In this study, a rotavirus species A detected from an outbreak of enteritis in a Mexican commercial rabbitry was genotypically characterized. Based on sequence and phylogenetic analysis of the VP7 and VP4 genes, the strain identified in this study (C-3/15) demonstrated a G3 P[8] genotype of rotavirus, which had not previously been reported in rabbits. Moreover, both genes were closely related to human, not lapine, rotaviruses. The G3 genotype has been reported in a wide variety of hosts, including humans and rabbits, whereas the P[8] genotype has only been reported in humans. Because this combination of genotypes has never been identified in rabbits, it is proposed that the finding presented here is possibly the result of an interspecies transmission event. This is the first work to study the molecular characteristics of rotaviruses in rabbits in Mexico, as well as the identification of human G3 and P[8] genotypes in a rabbit with enteric disease. [ABSTRACT FROM AUTHOR]

Copyright of Veterinarska Stanica is the property of Croatian Veterinary Institute and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Human astroviruses (HAstVs) were examined in 1625 stool samples collected from outpatient children with diarrhea who visited clinics in 4–6 prefectures in Japan. The study was conducted over a period of 4 years from July 2018 to June 2022, spanning the period before and after the emergence of COVID‐19. The HAstV and other diarrheal viruses including group A rotavirus, norovirus and sapovirus were screened by RT‐PCR. Of these, HAstV was detected in 140 out of 1625 (8.6%). When the stool samples were categorized by the year of collection, HAstV was detected in 2018–2019, 2019–2020, 2020–2021, and 2021–2022 with the prevalences of 3.1%, 6.6%, 3.0%, and 20.1%, respectively. Among 140 HAstV positive cases, HAstV1, MLB1, MLB2, HAstV3, MLB3, and VA2 were detected in 77, 46, 10, 3, 2, and 2 samples, respectively. High infection rate was found in children 1–3 years of age (95/140; 67.9%). Severity of the disease increased with the co‐infection with norovirus. During the surveillance of 2021–2022, two outbreaks of HAstV were detected, one was an outbreak of HAstV1 and MLB2 in September, 2021 in Kyoto, the second was an outbreak of HAstV1 and MLB1 in December, 2021 in Kyoto and Shizuoka. It was interesting to observe that mixed‐infection of astroviruses between HAstV1 and MLB1 was reported for the first time in this study. Characterization of the subgenotypes in HAstV1 and HAstV3 indicated that most HAstV1 circulating in Japan belonged to HAstV1a and only a single strain was HAstV1b, whereas all HAstV3 was identified as the HAstV3c subgenotype. In conclusion, several HAstV genotypes, including classic HAstV, novel MLB, and VA genotypes, were detected in this study. The incidence of astrovirus outbreaks was also reported after the pandemic of COVID‐19 in 2021–2022. [ABSTRACT FROM AUTHOR]

Group A rotaviruses (RVA) remain a principal cause of childhood diarrhea in the UAE, despite universal vaccine use. Monitoring genetic diversity is important for identifying prevalent genotypes and escape mutants. Although real‐time polymerase chain reaction (RT‐PCR) is widely used for RVA genotyping, it may not detect some new strains. This study evaluates nanopore sequencing and RT‐PCR for RVA genotyping. Thirty‐three RVA strains from children under 5 years presenting with diarrhea were genotyped using both methods. Thirteen strains were genotyped by RT‐PCR and confirmed by nanopore sequencing. Fifteen strains were genotyped by nanopore method alone. Most PCR‐genotyped strains (56%) had the VP7 G9 genotype, with G3 in five strains and G12 in two. For VP4, P8 (n = 8) and P4 (n = 7) were dominant. The most frequent combinations were G9P[8] (31%) and G9P[4] (25%). Nanopore sequencing of 28 strains revealed G3P[8] (29%) as the most prevalent, followed by G8P[8] (18%). G9P[8] and G2P[4] were present in 14% of samples with G12P[6] being the rarest (7%). Other combinations were detected in 4% the specimens with one nontypeable. Nanopore sequencing was superior to PCR in identifying diverse and emerging genotypes like G8P[8]. This method may enhance surveillance studies and guide preventive measures for RVA gastroenteritis. [ABSTRACT FROM AUTHOR]

Background: Viral enteric infections are illnesses caused by several types of viruses that primarily affect the gastrointestinal system. Globally, three major viruses associated with gastroenteritis, including norovirus (NoV), rotavirus A (RVA), and human adenovirus (HAdV), have been widely recognized. Accordingly, a rapid and sensitive diagnostic tool, such as immunochromatographic (IC) test, for diarrheal virus detection is required and helpful for rapid diagnosis. Methods: The IP-Triple I IC test kit for simultaneous triple virus detections of NoV, RVA, and HAdV was evaluated for its efficacy by using stool specimens that were known positive for these viruses by real-time RT-PCR or PCR, which was used as the gold standard method. Results: The results revealed that the IP-Triple I IC test kit exhibited a very high level of specificity (100%) for NoV, RVA, and HAdV detections, while the sensitivity was slightly different for these three viruses. The sensitivity of detection for RVA was 86.7%, whereas those for NoV and HAdV were 70.6% and 76.2%, respectively. This IPTriple I IC kit could detect common antigens of a wide range of NoV, RVA, and HAdV genotypes (NoV GII.2, GII.4, GII.6, GII.7, GII.10, GII.17, RVA G1P[8], G2P[4], G3P[8], G8P[8], G9P[8], HAdV-C1, -C2, -C5, -A12, -F40, and -F41). Conclusions: Our results revealed that the IP-Triple I IC test kit is highly effective for simultaneous, direct detection of common antigens of several genotypes of NoV, RVA, and HAdV in stool specimens and useful for screening and rapid diagnosis of diarrheal viruses during the seasonal outbreak. [ABSTRACT FROM AUTHOR]

Background: Enteric infections are hypothesized to be associated with intussusception in children. A small increase in intussusception following rotavirus vaccination has been seen in some settings. We conducted post-marketing surveillance for intussusception following rotavirus vaccine, Rotavac introduction in India and evaluated association of intussusception with enteric pathogens. Methods: In a case-control study nested within a large sentinel hospital-based surveillance program in India, stool samples from 272 children aged less than 2 years admitted for intussusception and 272 age-, gender- and location-matched controls were evaluated with Taqman array card based molecular assays to detect enteric viruses, bacterial enteropathogens and parasites. Matched case-control analysis with conditional logistic regression evaluated association of enteropathogens with intussusception. Population attributable fractions (PAF) were calculated for enteropathogens significantly associated with intussusception. Results: The most prevalent enteropathogens in cases and controls were enteroaggregative Escherichia coli, adenovirus 40/41, adenovirus C serotypes and enteroviruses. Children with intussusception were more likely to harbor adenovirus C serotypes (adjusted odds-ratio (aOR) = 1.74; 95% confidence interval (CI) 1.06–2.87) and enteroviruses (aOR = 1.77; 95% CI 1.05–2.97) than controls. Rotavirus was not associated with increased intussusception risk. Adenovirus C (PAF = 16.9%; 95% CI 4.7% − 27.6%) and enteroviruses (PAF = 14.7%; 95% CI 4.2% − 24.1%) had the highest population attributable fraction for intussusception. Conclusion: Adenovirus C serotypes and enteroviruses were significantly associated with intussusception in Indian children. Rotavirus was not associated with risk of intussusception. [ABSTRACT FROM AUTHOR]

Objective To assess the presence of enteroviruses and diarrheal viruses in the environmental sewage of Aba in 2022, and to provide evidence supporting effective control measures against these viruses. Methods Sewage samples were collected monthly throughout 2022, then eluted and concentrated by anionic membrane adsorption. Through this, enteroviruses were isolated and identified. At the same time, nucleic acid was extracted from the concentrated liquid, and real-time fluorescent PCR was used to detect several diarrhea viruses, such as group A rotavirus, norovirus, Sapovirus, human astrovirus and enteric adenovirus. Results A total of 12 environmental sewage samples were collected from January to December 2022. Out of these, 3 strains of polioviruses and 9 non-polio enteroviruses were isolated. All 3 poliovirus strains were detected as Type 3 vaccine-like poliovirus (PV) strains. Group A rotavirus and norovirus genotype II were detected every month. Norovirus type I and enteric adenovirus were observed in all months except November. Human astrovirus was absent in samples from July, August, November and December. Sapovirus was not identified throughout the year. Conclusion Enteroviruses and diarrheal viruses exist widely in Aba's environmental sewage. Environmental sewage surveillance can offer a comprehensive view of these viruses' etiological and epidemiological characteristics. Continuous environmental sewage surveillance can predict and warn the epidemic and outbreak of enteroviruses and diarrheal virus-related diseases. [ABSTRACT FROM AUTHOR]

Most human rotaviruses belong to the Wa-like, DS-1-like, or AU-1-like genotype constellation. The AU-1-like constellation, albeit minor, captured attention because its prototype strain AU-1 originated from feline rotavirus, leading to the concept of interspecies transmission of rotavirus. The AU-1 genome sequence determined by various laboratories over the years has documented two conflicting VP7 sequences in the GenBank. As culture-adaptation may introduce changes in the viral genome, the original fecal (wild-type) and the seed stock of culture-adapted AU-1 genomes were sequenced using the Illumina's MiSeq platform to determine the authentic AU-1 sequence and to identify what mutational changes were selected during cell-culture adaptation. The wild-type and culture-adapted AU-1 genomes were identical except for one VP4-P475L substitution. Their VP7 gene was 99.9% identical to the previously reported AU-1 VP7 under accession number AB792641 but only 92.5% to that under accession number D86271. Thus, the wild-type sequences determined in this study (accession numbers OR727616-OR727626) should be used as the reference. The VP4-P475L mutation was more likely incidental than inevitable during cell-culture adaptation. This was the first study in which the whole genomes of both wild-type and cultured RVA strains were simultaneously determined by deep sequencing. [ABSTRACT FROM AUTHOR]

Rotavirus A (RVA) is the main cause of acute gastroenteritis among children under the age of five globally. The unusual bat‐like human RVA strains G3P[10] (RVA/Human‐wt/THA/CMH079/05/2005/G3P[10] and RVA/Human‐wt/THA/CMH‐S015‐19/2019/G3P[10]) were detected in children with acute gastroenteritis in 2005 and 2019, respectively, in the same geographical area of Northern Thailand. To elucidate the genetic backgrounds of these unusual or bat‐like human RVA strains, the complete genome of these RVA strains was sequenced and phylogenetically analyzed. All eleven genome segments of these G3P[10] strains were genotyped as G3‐P[10]‐I8‐R3‐C3‐M3‐A9‐N3‐T3‐E3‐H6, which is closely related to bat G3P[10] RVA strain (RVA/Bat‐tc/CHN/MYAS33/2013/G3P[10]) and bat‐like human RVA strain (RVA/Human‐wt/THA/MS2015‐1‐0001/2015/G3P[10]). The findings indicate that human G3P[10] RVA strains detected in this study (RVA/Human‐wt/THA/CMH079/05/2005/G3P[10] and RVA/Human‐wt/THA/CMH‐S015‐19/2019/G3P[10]) contained all eleven genome segments similar to those of bat RVA strains and appeared to be human RVA strains of bat origin. Phylogenetic analysis revealed that several genome segments of these two RVA strains were also closely related with those of other species in addition to bats and had a zoonotic transmission history. The results of this study supported the roles of interspecies transmission of RVA strains among bats and humans in the natural environment and provided convincing evidence that the evolution of human RVAs was closely interrelated with those of animal RVAs. Continuing surveillance of RVAs in humans and animals is imperative to gain a better understanding of the origin and the evolution of these viruses. [ABSTRACT FROM AUTHOR]

Introduction: Bovine torovirus (BToV), Bovine enterovirus (BEV), Bovine norovirus (BNoV), Bovine coronavirus (BCoV), Bovine rotavirus (BRV), and Bovine viral diarrhea virus (BVDV) are significant pathogens causing diarrhea in calves, characterized by their high prevalence and challenging prevention and control measures. Methods: We analyzed 295 calf diarrhea samples, amplifying the M gene from BToV-positive samples, the 5'UTR gene from BEV-positive samples, the RdRp gene from BNoV-positive samples, the VP7 gene from BRV-positive samples, the S gene from BCoV-positive samples, and the 5'UTR gene from BVDV-positive samples. Subsequent homology analysis and phylogenetic tree construction were performed. Results: The overall viral positive rate in Guangdong Province was 21.36%. Specific detection rates were as follows: Foshan City at 50.00% (18/36), Guangzhou City at 43.90% (36/82), Huizhou City at 21.21% (7/33), Yangjiang City at 2.08% (1/48), Meizhou City at 1.39% (1/72), and Heyuan City at 0.00% (0/24). The detection rates for BToV, BEV, BNoV, BCoV, BRV, and BVDV were 0.34% (1/295), 6.10% (18/295), 0.68% (2/295), 1.36% (4/295), 10.85% (32/295), and 2.03% (6/295), respectively. Notably, the highest overall virus detection rate was observed in the Guangzhou-Foshan region, with BRV and BEV showing the highest detection rates among the six viruses. This study marks the first report of BToV and BNoV in Guangdong Province. Phylogenetic analysis revealed that the BToV strain belonged to type II, sharing genetic similarities with epidemic strains from various provinces in China. The BEV strains were categorized into E and F types, with the F type being the predominant strain in Guangdong Province and exhibiting the closest genetic relationship to strains from Heilongjiang and Guangxi. The BNoV strains, along with Hebei strains, were identified as GIII.2 subgenotype. BCoV strains showed the highest genetic similarity to strains from Sichuan. All BRV strains were classified under the G6 subtype and had the closest genetic relationship with human rotavirus strains. BVDV strains were identified as subtype 1b, closely related to the Beijing strain. In conclusion, this study investigated the prevalence and evolutionary characteristics of diarrheaassociated viruses in calves in specific areas of Guangdong Province, providing a valuable reference for establishing effective prevention and control measures in cattle farms. [ABSTRACT FROM AUTHOR]

The purpose of this review is to present modern data on the correlations between viruses and celiac disease. Some viruses probably have crucial role in celiac disease pathogenesis according to the results of recent studies. However, the virus exposure itself does not lead to the development of autoimmune disease. Number of studies have demonstrated that there are several viruses that can cause celiac disease, and several activation pathways leading to food tolerance loss. Thus far, the role of some reoviruses strains has been proven in the development of celiac disease. Further research is needed for better understanding of the viruses role in the celiac disease development and to improve management and prevention of this disease.

Background: This hospital-based cross-sectional study aims to investigate the epidemiologic and clinical characteristics of rotavirus group A (RVA) infection among children with acute gastroenteritis and to detect the most common G and P genotypes in Egypt. Methods: A total of 92 stool samples were collected from children under five who were diagnosed with acute gastroenteritis. RVA in stool samples was identified using ELISA and nested RT-PCR. Common G and P genotypes were identified utilizing multiplex nested RT-PCR assays. Results: RVA was detected at a rate of 24% (22 /92) using ELISA and 26.1% (24 /92) using VP6 nested RT-PCR. The ELISA test demonstrated diagnostic sensitivity, specificity, and accuracy of 91.7%, 100%, and 97.8%, respectively. G3 was the most prevalent G type (37.5%), followed by G1 (12.5%), whereas the most commonly detected P type were P[8] (41.7%) and P[6] (8.2%). RVA-positive samples were significantly associated with younger aged children (p = 0.026), and bottle-fed (p = 0.033) children. In addition, RVA-positive samples were more common during cooler seasons (p = 0.0001). Children with rotaviral gastroenteritis had significantly more frequent episodes of diarrhea (10.87 ± 3.63 times/day) and vomiting (8.79 ± 3.57 times/day) per day (p = 0.013 and p = 0.011, respectively). Moreover, they had a more severe Vesikari clinical score (p = 0.049). Conclusion: RVA is a prevalent cause of acute gastroenteritis among Egyptian children in our locality. The discovery of various RVA genotypes in the local population, as well as the identification of common G and P untypeable strains, highlights the significance of implementing the rotavirus vaccine in Egyptian national immunization programs accompanied by continuous monitoring of strains. [ABSTRACT FROM AUTHOR]

Group A rotaviruses (RVAs) are widespread in humans and many animal species and represent the most epidemiologically important rotavirus group. The aim of the study was the identification of the genotype pattern of human RVA strains circulating in Poland, assessment of their phylogenetic relationships to pig RVAs and identification of reassortant and zoonotic virus strains. Human stool samples which were RVA positive (n = 166) were collected from children and adults at the age of 1 month to 74 years with symptoms of diarrhoea. Identification of the G and P genotypes of human RVAs as well as the complete genotype of reassortant and zoonotic virus strains was performed by the use of an RT‐PCR method. The G (G1–G4, G8 or G9) and/or P (P[4], P[6], P[8] or P[9]) genotypes were determined for 148 (89.2%) out of 166 RVA strains present in human stool. G1P[8] RVA strains prevailed, and G4P[8] (20.5%), G9P[8] (15.7%) and G2P[4] (13.3%) human RVA strains were also frequently identified. The full genome analysis of human G4P[6] as well as pig G1P[8] and G5P[6] RVAs revealed the occurrence of porcine–human reassortants and zoonotic RVAs. Detection of G4P[6] in pigs confirms their role as a reservoir of zoonotic RVAs. [ABSTRACT FROM AUTHOR]

Human noroviruses (HuNoVs), the most prevalent viral contaminant in food, account for a substantial proportion of nonbacterial gastroenteritis cases. Extensive work has been focused on the diagnosis of HuNoVs in clinical samples, whereas the availability of sensitive detection methods for their detection in food is lacking. Here, we developed a virus enrichment approach utilizing graphene-based nanocomposites (CTAB-rGO-Fe3O4) that does not rely on large instruments and is suitable for on-site food pretreatment. The recovery efficiency of the developed virus enrichment procedure for serially diluted GII.4 norovirus ranged from 10.06 to 72.67% in strawberries and from 2.66 to 79.65% in oysters. Furthermore, we developed a real-time recombinase polymerase amplification (real-time RPA) assay, which can detect as low as 1.22 genome copies µL−1 of recombinant plasmid standard and has no cross-reactivity with genomes of astrovirus, rotavirus, adenovirus, and MS2 bacteriophage. Notably, the combined virus enrichment and real-time RPA detection assay enhanced the detection limits to 2.84 and 37.5 genome copies g−1 in strawberries and oysters, respectively, compared to those of qPCR. Our strategy, the graphene-based virus enrichment method combined with real-time RPA, presents a promising tool for sensitively detecting HuNoVs in food samples. [ABSTRACT FROM AUTHOR]

Background: Acute infectious enteritis is one of the most common causes of death among children in developing countries. In developed countries, 75% to 90% of childhood acute gastroenteritis is caused by viruses, and approximately 20% is caused by bacteria. It is known that parasitic agents are responsible for approximately 5-10% of acute infectious gastroenteritis. Metronidazole, a nitroimidazole, is restricted to Clostridium difficile-associated colitis, anaerobic bacteria, protozoans (such as Entamoeba histolytica, Giardia intestinalis, Trichomonas vaginalis) and microaerophiles (such as Helicobacter pylori). The use of metronidazole in patients with enteritis without a microbiological diagnosis may lead to increased antibiotic resistance in bacteria, unnecessary costs, adverse events and intestinal dysbiosis. Materials and Methods: In our hospital, all stool samples examined from patients under 18 years of age in 2020 and 2021 were tested using FilmArray GI Panel (Biomerioux, France), direct microscopy of stool, rapid antigen tests (Rotavirus, Adenovirus, G. intestinalis, Cryptosporidium spp, C. difficile toxin A/B ( CerTest Biotech, Spain)), E. histolytica microELISA (Adhesin Ag, Entamoeba CELISA Path, Australia) results and stool cultures were scanned retrospectively. Metronidazole usage status of the patients was obtained from the Hospital Information Management System. Results: With the diagnostic methods used in this study, stool samples of 2282 patients were evaluated over a two-year period and 782 factors were detected in 663 (29%) patients. The most frequently detected agent was Rotavirus with a rate of 11.6%. There were 564 (24.7%) patients with a single agent, 77 (3.3%) patients with two agents, and 22 (0.9%) patients with three agents. While 124 (18.7%) of the patients used metronidazole, no factor requiring the use of this drug was detected in 47 (7%) of them. Conclusions: As a result, detecting the microorganisms that cause gastroenteritis with appropriate diagnostic methods will contribute to patients receiving appropriate treatment and preventing unnecessary antibiotic use and antibiotic resistance. [ABSTRACT FROM AUTHOR]

Immunoglobulins Y (IgY) purified from egg yolks of hens represents an attractive, cost-effective alternative for the development of new diagnostic and therapeutic platforms. In this study, we evaluated the therapeutic efficacy of rotavirus-specific IgY in a cynomolgus monkey (Macaca fascicularis) model. Animals were experimentally infected with human rotavirus Group A (RVA), the most common cause of severe acute diarrhoea among young children worldwide. Animals were administered human RVA (3.1 × 107 FFU/mL) by oral gavage, challenged with 2.5 mg of anti-RVA IgY orally, and monitored for five days according to clinical, haematological and biochemical parameters; serum electrolyte levels; viral shedding; and histopathological changes. Immunotherapy with anti-RVA IgY had a protective effect against severe rotavirus-induced enteritis in four of the ten treated monkeys, as evidenced by histopathological findings. Although only one animal had diarrhoea, all but one exhibited virus shedding regardless of the treatment. [ABSTRACT FROM AUTHOR]

Rotaviruses (RVs) are classified into nine species, A–D and F–J, with species A being the most studied. In rotavirus of species A (RVA), replication occurs in viroplasms, which are cytosolic globular inclusions composed of main building block proteins NSP5, NSP2, and VP2. The co-expression of NSP5 with either NSP2 or VP2 in uninfected cells leads to the formation of viroplasm-like structures (VLSs). Although morphologically identical to viroplasms, VLSs do not produce viral progeny but serve as excellent tools for studying complex viroplasms. A knowledge gap exists regarding non-RVA viroplasms due to the lack of specific antibodies and suitable cell culture systems. In this study, we explored the ability of NSP5 and NSP2 from non-RVA species to form VLSs. The co-expression of these two proteins led to globular VLSs in RV species A, B, D, F, G, and I, while RVC formed filamentous VLSs. The co-expression of NSP5 and NSP2 of RV species H and J did not result in VLS formation. Interestingly, NSP5 of all RV species self-oligomerizes, with the ordered C-terminal region, termed the tail, being necessary for self-oligomerization of RV species A–C and G–J. Except for NSP5 from RVJ, all NSP5 interacted with their cognate NSP2. We also found that interspecies VLS are formed between closely related RV species B with G and D with F. Additionally, VLS from RVH and RVJ formed when the tail of NSP5 RVH and RVJ was replaced by the tail of NSP5 from RVA and co-expressed with their respective NSP2. IMPORTANCE Rotaviruses (RVs) are classified into nine species, A–D and F–J, infecting mammals and birds. Due to the lack of research tools, all cumulative knowledge on RV replication is based on RV species A (RVA). The RV replication compartments are globular cytosolic structures named viroplasms, which have only been identified in RV species A. In this study, we examined the formation of viroplasm-like structures (VLSs) by the co-expression of NSP5 with NSP2 across RV species A to J. Globular VLSs formed for RV species A, B, D, F, G, and I, while RV species C formed filamentous structures. The RV species H and J did not form VLS with their cognates NSP5 and NSP2. Similar to RVA, NSP5 self-oligomerizes in all RV species, which is required for VLS formation. This study provides basic knowledge of the non-RVA replication mechanisms, which could help develop strategies to halt virus infection across RV species. [ABSTRACT FROM AUTHOR]

Identified as a potential reference pathogen by the WHO Guidelines for Drinking-Water Quality, Rotavirus (RV) is among the main enteric viruses that cause waterborne diseases. The aim of this study was to identify and correlate the presence of RV in collective and individual water sources of rural communities in the state of Goiás, within the seasons in which the collections were made (rainy and dry seasons). For this, 86 water samples in the dry period and 160 samples in the rainy period were collected. Concentration of water samples, extraction of viral genetic material and molecular tests were performed. When analyzing the presence of RV in the samples, taking into consideration the period studied, RV was found to be more prevalent in the dry season (54.7%) than in the rainy season (20%), showing a strong statistical association with the dry season (p-value < 0.001). The presence of pathogenic microorganisms in water is a public risk issue, enabling the emergence of outbreaks, endemics and epidemics. In the present research, there was an association between the presence of Rotavirus and the dry period of the year when compared to the rainy period. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine rotavirus re-emerged in recent years in China. In this study, we analyzed the genomic characterizations of six porcine rotavirus. A strains (PoRVA) isolated from three sow farms in a company at different sampling periods, and in detail dissected their evolutionary relationship. Our results showed transmission and recombination happened frequently within and between pig farms under the same production system. More importantly, several gene segments of isolates came from human rotavirus strains, suggesting the potential cross–species infection between humans and pigs. Our findings provide insights into the transmission and evolution of porcine rotavirus (PoRV) between farms and enrich the molecular epidemiological data of porcine rotavirus in China. Porcine rotavirus (PoRV) is a significant enteric pathogen causing gastroenteritis in piglets, which causes huge economic loss to the Chinese pig industry. In this study, six porcine rotavirus A strains were isolated from three adjacent sow farms belonging to the same company within one year, which suffered severe diarrhea outbreaks. AHBZ2303 (G11P[7]) and AHBZ2305 (G9P[23]), AHBZ2304 (G9P[23]) and AHBZ2312 (G4P[6]), AHBZ2310 (G9P[23]) and AHBZ2402 (G5P[23]) were isolated from Farm A, B and C, respectively. All six isolates were related to human rotavirus through complete genome analysis, suggesting the potential cross–species infection between humans and pigs. Evolutionary analysis revealed that AHBZ2303 and AHBZ2304 likely emerged simultaneously in Farm A and B, and then AHBZ2304 was introduced to Farm A and C, leading to the emergence of AHBZ2305 and AHBZ2310. Recombination and large variation were identified for AHBZ2312 and AHBZ2402. These findings provided insights into the transmission and evolution of PoRV among farms and underscored the need for enhanced monitoring to mitigate the risk of outbreaks from novel variants. [ABSTRACT FROM AUTHOR]

A total of 490 diarrhoeic samples from calves aged between 0 and 6 months were screened for the presence of different G- and P-genotypes of rotavirus circulating in bovines in the Kashmir Valley. Of the 490 diarrhoeic samples, Group A rotavirus was detected in 68 (13.87%) samples by polymerase chain reaction (PCR) followed by RNA-PAGE. Genotyping analysis revealed G10, G6, G3, P[11] and P[5] to be the predominant types. The most common types of combinations detected were G10P[11] (27.90%) and G6P[11] (20.60%). The prevalence rate of G10 and P[11] decreased from 60% to 36.76% and 100%–69.11%, respectively. Genotypes G6, G3, P[1] and P[5], which were not previously reported, were detected and unusual combinations such as G6P[11], G3P[11], G10P[5], G3P[5], G6P[1], G6P[5], G6+G8P[11] were also observed for the first time. Fluctuations in the predominant types, emergence of new types and possible genetic reassortment events suggest an unstable epidemiological situation and the need for continuous surveillance of the circulating types to ensure the suitability of the vaccination programme. The present data suggests G10, G6, P[11] and P[5] genotypes could be incorporated in the polyvalent vaccine to offer increased protection against bovine rotavirus infection in India. [ABSTRACT FROM AUTHOR]

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Introduction: Rotaviruses A (RVA) are a major cause of acute viral gastroenteritis in humans worldwide and are responsible for about two million hospitalizations per year. They can also infect other mammals such as pigs, calves, goats, lambs, and horses, in which they are also considered a major cause of viral diarrhea. While RVA is well studied in humans and domestic animals, its occurrence in wild ruminants is not well known. The RVA genome is a double-stranded RNA consisting of 11 segments, and genotyping is based on the VP7 (G) and VP4 (P) segments. Currently, there are 42G genotypes and 58P genotypes. RVA has a high mutation rate, and some combinations of G and P genotypes can infect different animal species, leading to speculation about the potential for zoonotic transmission. Materials and methods: A total of 432 fecal samples were collected from roe deer, red deer, chamois, mouflon and Alpine ibex in Slovenia between 2017 and 2021. To investigate the presence of RVA in wild ruminants, real-time RT-PCR was used. Positive samples were subjected to next generation sequencing (NGS) using RIP-seq method. Results and discussion: In total, 7 samples were RVA positive. Complete genomes were determined and phylogenetically analyzed for all 7 RVAs. Four different genotype constellations were present in 7 positive RVA animals: G8- P[14]-I2- R2-C2-M2-A3-N2-T6-E2-H3, G6-P [14]-I2-R2-C2-M2-A11-N2-T6- E2-H3, G10-P [15]-I2-R2-C2-M2-A3-N2-T6-E2-H3 and G10-P [15]-I2-R2-C2- M2-A11- N2-T6-E2-H3. Genotypes G6P[14] and G10P[15] were found in both roe deer and red deer, representing the first confirmed occurrence of RVA in red deer. In addition, genotype G8P[14] was found in chamois, representing the first known case of positive RVA in this species. Some of these genotypes have also been found in humans, indicating the potential for zoonotic transmission. [ABSTRACT FROM AUTHOR]