Molecular Characterization of Bovine Rotaviruses and Coronaviruses in Diarrheic Calves in Egypt (2014-2019)

Bovine Rotaviruses group A (BRVA) and Bovine Coronaviruses (BCoV) are the most prevalent viral agent worldwide in diarrheic calves aged less than 6 weeks, causing economic losses due to retarded growth, increased susceptibility to other infections, treatment cost, and calf mortalities. This study aimed to detect and molecularly characterize BRVA and BCoV from diarrhetic calves. A total of 82 fecal samples were collected from calves aged less than one month from three Egyptian governorates (Alexandria, Ismailia, and Sharqia). All fecal samples were tested for BRVA and BCoV by using probe based quantitative reverse-transcription polymerase chain reaction (qRT-PCR). Consequently, positive samples contain relatively high viral genomic load were examined by RT-PCR for amplification of viral protein 7 (VP7) and viral protein 4 (VP4) genes (G- and P- typing) for BRV and full length S1 gene for BCoV. Out of 82 of tested samples, 14 (17.1%) and 22 (26.8%) were positive by qRT-PCR for BRV and BCoV, respectively. Only three and five samples had relatively high genomic load for BRV and BCoV, respectively for further testing by RT-PCR. BRV G-type was found in two samples and P-type was detected in one sample. The sequence analysis and phylogenetic tree typed these positive samples as P11 and G10. The sequences and phylogenetic analysis of BCoV positive strains (n=5) showed closely related viruses to each other and similar to previously characterized strains in Egypt since 2014. Further studies are required to antigenically characterize the circulating BRV and BCoV in Egypt.

Protein losing enteropathy (PLE) secondary to norovirus infection in a 4 year old patient

Case Report: Protein losing enteropathy (PLE) occurs when proteins leak from the gastrointestinal (GI) system more rapidly than they are produced. Inflammation of the GI tract facilitates increased membrane permeability of gastric mucosa, leading to excess protein leakage. 1 PLE in children has been associated with CMV, rotavirus, COVID-19, HIV, C. difficile, and autoimmune diseases like Crohn's Disease. 2-6 Norovirus is a known cause of PLE in immunocompromised pediatric patients. 7-8 However, to our knowledge, there are no case reports about PLE precipitated by norovirus in immunocompetent pediatric patients. The purpose of this case report is to present a case of PLE precipitated by a norovirus infection in a 4- year-old previously healthy child. While the above gastrointestinal viruses have been proposed as precipitators for this disease, PLE precipitated by norovirus infection has not been well described. This case also highlights the importance of early diagnosis and management to avoid complications. Method(s): Our patient initially presented with two days of abdominal pain, diarrhea, emesis, reduced urine output, and swelling of the lower extremities. He was exposed to several sick family members-his sister had upper respiratory symptoms and his grandmother had gastrointestinal symptoms. Physical exam was notable for diminished breath sounds in the right lower lobe, abdominal distension with diffuse tenderness and dullness to percussion, significant scrotal and penile edema, and bilateral lower extremity pitting edema. Laboratory results revealed leukocytosis, hypoalbuminemia, hyponatremia, elevated aspartate aminotransferase (AST), and elevated serum alpha-1-antitrypsin, as well as low Immunoglobulins G and M. CD3 and CD4 levels were low reflecting cellular immune dysregulation seen in patients with PLE. IgA and Tissue Transglutaminase (TTF) were within normal limits. Ebstein Barr Virus and cytomegalovirus IgM antibodies were negative. COVID IgG was negative as well. His Polymerase chain reaction (PCR) gastrointestinal panel was positive for norovirus. A chest X-ray showed a large right pleural effusion. Abdominal CT revealed large ascites slightly more predominant in the upper abdomen, mesenteric lymphadenitis, and bilateral pleural effusions. Echocardiogram showed small anterior and apical pericardial effusions. Result(s): Based on the patient's elevated serum alpha-1 antitrypsin levels, hypoalbuminemia, low levels of immunoglobulins and lymphocytes, and clinical manifestations of ascites, bilateral pleural effusions, pericardial effusion, and dependent edema, along with a positive PCR for norovirus, the diagnosis of PLE secondary to Norovirus was made. Conclusion(s): This case demonstrates the importance of recognizing viruses like Norovirus as potential causes of PLE to avoid a delay in diagnosis and initiation of therapy, and to avoid unnecessary additional testing. Copyright © 2023 Southern Society for Clinical Investigation.

Dynamics of the frequency of hospitalizations of children under 5 years of age with rotavirus infection in the absence of universal vaccination against rotavirus (15-year follow-up)

Purpose - to monitor the dynamics of the frequency of hospitalization of children under 5 years of age with rotavirus infection during 15 years in the absence of universal vaccination and to determine the burden of rotavirus infection and the feasibility of introducing a rotavirus vaccine into the National Vaccination Calendar. Materials and methods. Active surveillance of rotavirus infection among hospitalized children under 5 years of age was conducted according to a standard protocol prepared by WHO for the global rotavirus infection surveillance network. A comparison was made of the frequency of hospitalization of children under 5 years of age with rotavirus acute gastroenteritis (AGE) by sentinel years. Stool samples were collected within 2 days of hospitalization and sent to the virology laboratory for testing. Detection of rotavirus was carried out using ELISA. Genotyping of rotaviruses was carried out in WHO laboratories by the method of multiplex RT-PCR. Results. During 15 years (2007-2021) of observation, there were 7,821 children underthe age of 5 who were hospitalized in connection with an acute intestinal infection at the Kyiv Surveillance Center. Rotavirus infection was determined in 3387 children (43.3%). A consistently high frequency of hospitalizations with rotavirus AGE was observed with a slight downward trend over the years (from 50-75% in the first years to 33-39% in 2018-2019). In 2021 (the 15thyear of observation), the average annual frequency of rotavirus AGE decreased even more significantly and amounted to 21 % (a 61 % decrease compared to the first years of oPservation). The unexpected decrease in the frequency of rotavirus diarrhea among hospitalized children in the 15thyear of observation in the absence of universal vaccination against rotavirus is difficult to explain. A number of factors could have contributed to the above-mentioned decrease, the most likely of which could be the coincidence in time with the endemic of COVID-19, in which anti-ebidemic measures are similar to those for intestinal infections. While the average annual freguency of hospitalizations with rotavirus infection decreased, the frequency of hospitalizations remained high in certain months of the year, starting from January to June, which increased the Purden on inpatients in certain, mostly cold, months of the year. Further active monitoring of rotavirus infection is necessary to definitively resolve the issue of whether the decrease in the frequency of rotavirus infection by the 15thyear is temporary or is a natural course of the epidemic process and will continue to occur. Conclusions. In Ukraine, in the absence of universal vaccination against rotavirus, during the 15-year (2007-2021) observation period, a consistently high frequency of rotavirus infection was noted among hospitalized children under 5 years of age with AGE, with a slight downward trend overthe years. In the 15thyear of follow-up (2021), there was an unusual decline in the proportion of rotavirus infection among hospitalized children, which may have been contributed to by a number of factors, the most likely of which could be the coincidence in the period with the COVID-19 epidemic, in which anti-epidemic measures are similar to those for intestinal infections. Even with a decrease in the average annual frequency of rotavirus AGE in the 15thyear of observation, a consistently high rate of hospitalization with rotavirus gastroenteritis remains in certain months of the year, which increases the burden on inpatients. The significant burden of rotavirus infection among children under the age of 5 years old determines the expediency of including the rotavirus vaccine in the National Vaccination Calendar of Ukraine. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of all participating institutions. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors. © 2022 The authors

Evidence on the Preventive Effects of the Postbiotic Derived from Cow's Milk Fermentation with Lacticaseibacillus paracasei CBA L74 against Pediatric Gastrointestinal Infections.

Postbiotics are commonly defined as preparations of inanimate probiotics and/or their cellular components and/or their metabolites/end products that confer health benefits on the host. They have been suggested as a promising strategy to limit infectious diseases. Emerging evidence support the efficacy of the postbiotic derived from cow's milk fermentation with the probiotic Lacticaseibacillus paracasei CBAL74 (FM-CBAL74) in preventing pediatric infectious diseases. We aimed at reviewing the evidence available.

ASVAC2022 : 8th Asian Vaccine Conference.

Aiming to further the Immunization Partners in Asia Pacific (IPAP)'s vision of a world where no one suffers from a vaccine preventable disease, the 8th Asian Vaccine Conference (ASVAC 2022) was held in Colombo, Sri Lanka and virtually from 15 to 18, September 2022 (www.asianvaccine.com). This conference followed those held in Siem Reap, Cambodia (2009), Manila, Philippines (2010), Jakarta, Indonesia (2011), Cebu, Philippines (2013), Hanoi, Vietnam (2015), Singapore (2017) and Naypyidaw and Yangon, Myanmar (2019). The ASVAC2022 themed "Immunization: in Era of Pandemics," commenced with the EPI Managers' Workshop, followed by pre-conference workshops and Vaccinology Masterclass, followed by the main conference featuring 5 plenary lectures, 6 partner-led symposia, free paper and poster presentations, and industry-supported lunch and evening sessions. There were over 1830 registered participants, with 112 attending in person and 998 virtually from 63 countries. The conference was organized by IPAP and hosted by the Vaccine and Infectious Disease Forum of Sri Lanka, Sri Lanka College of Pediatricians, Sri Lanka College of Microbiologists and College of General Practitioners of Sri Lanka, with the support of the Ministry of Health, Sri Lanka. The 9th ASVAC is scheduled to be held in Davao City, Philippines in late 2023.

Gastrointestinal organoids in the study of viral infections.

Viruses are among the most prevalent enteric pathogens. While virologists historically relied on cell lines and animal models, human intestinal organoids (HIOs) continue to grow in popularity. HIOs are non-transformed, stem cell derived, ex vivo cell cultures that maintain the cell type diversity of the intestinal epithelium. They offer higher throughput than standard animal models while more accurately mimicking the native tissue of infection than transformed cell lines. Here, we review recent literature that highlights virological advances facilitated by HIOs. We discuss the variations and limitations of HIOs, but also how HIOs have allowed for the cultivation of previously uncultivatable viruses and how they have offered insight into tropism, entry, replication kinetics, and host-pathogen interactions. In each case, we discuss exemplary viruses and archetypal studies. We discuss how the speed and flexibility of HIO-based studies contributed to our knowledge of SARS-CoV-2 and anti-viral therapeutics. Finally, we discuss current limitations of HIOs and future directions to overcome these.

The Value of Rotavirus Vaccination in Europe: A Call for Action.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has pushed many healthcare systems into crisis. High vaccine coverage amongst children reduces emergency room presentations, hospital admissions and deaths due to vaccine-preventable diseases, freeing up healthcare resources including polymerase chain reaction testing for patients with SARS-CoV-2. In Europe, rotavirus gastroenteritis leads to 75,000-150,000 hospitalisations and up to 600,000 medical encounters annually. Nevertheless, in 2022, only 18 countries in Europe (out of 38) have a publicly funded routine universal mass immunisation programme against rotavirus gastroenteritis. Evidence available in the last few years re-emphasises that rotavirus vaccines currently available in Europe are highly effective, preventing up to 96% of rotavirus-related hospitalisations in children less than 1 year of age (potentially 72,000-144,000 hospitalisations Europe-wide). Long-term surveillance indicates that rotavirus vaccination does not result in an overall increase in intussusception. On the contrary, increasing evidence suggests an overall reduction in intussusception in the first 12 months of life when early, high rotavirus vaccine coverage is achieved. Prevention of rotavirus gastroenteritis has marked positive impacts on parental wages and government tax revenue, with benefits extending across the whole economy. In the SARS-CoV-2 pandemic setting there is a new imperative to achieve high levels of paediatric vaccination against vaccine-preventable diseases, including rotavirus gastroenteritis. The introduction of rotavirus universal mass vaccination can be expected to reduce the number of preventable illnesses, hospitalisations and deaths caused by rotavirus gastroenteritis. Reducing vaccine-preventable diseases is particularly urgent at this time when healthcare systems are preoccupied and overwhelmed with SARS-CoV-2. Graphical abstract available for this article.

Australian Rotavirus Surveillance Program: Annual Report, 2021.

Abstract: This report from the Australian Rotavirus Surveillance Program describes the circulating rotavirus genotypes identified in children and adults during the period 1 January to 31 December 2021. During this period, 521 faecal specimens had been referred for rotavirus G- and P- genotype analysis, of which 474 were confirmed as rotavirus positive. Of these, 336/474 were wildtype rotavirus strains and 138/474 were identified as vaccine-like. Of the 336 wildtype samples, 87.5% (n = 294/336) were identified as G8P[8], and were detected in five of the six jurisdictions that provided samples for the reporting period. Two rotavirus outbreaks, located in the Northern Territory and Western Australia, were also attributed to G8P[8]. As with the 2020 reporting period, a low number of stool samples were received for this reporting period as a result of the COVID-19 pandemic. However, an unexpectedly high proportion of samples with unusual genotypes were identified which were potentially zoonotic in nature, including feline G3, P[9], bovine-like G8, P[14], and porcine-like G4, G6, P[1], and P[6]. Ongoing rotavirus surveillance is crucial to identify changes in genotypic patterns and to provide diagnostic laboratories with quality assurance by reporting incidences of wildtype, vaccine-like, or false positive rotavirus results.

Gastrointestinal organoids in the study of viral infections.

Viruses are among the most prevalent enteric pathogens. While virologists historically relied on cell lines and animal models, human intestinal organoids (HIOs) continue to grow in popularity. HIOs are non-transformed, stem cell derived, ex vivo cell cultures that maintain the cell type diversity of the intestinal epithelium. They offer higher throughput than standard animal models while more accurately mimicking the native tissue of infection than transformed cell lines. Here, we review recent literature that highlights virological advances facilitated by HIOs. We discuss the variations and limitations of HIOs, but also how HIOs have allowed for the cultivation of previously uncultivatable viruses and how they have offered insight into tropism, entry, replication kinetics, and host-pathogen interactions. In each case, we discuss exemplary viruses and archetypal studies. We discuss how the speed and flexibility of HIO-based studies contributed to our knowledge of SARS-CoV-2 and anti-viral therapeutics. Finally, we discuss current limitations of HIOs and future directions to overcome these.

The Value of Rotavirus Vaccination in Europe: A Call for Action.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has pushed many healthcare systems into crisis. High vaccine coverage amongst children reduces emergency room presentations, hospital admissions and deaths due to vaccine-preventable diseases, freeing up healthcare resources including polymerase chain reaction testing for patients with SARS-CoV-2. In Europe, rotavirus gastroenteritis leads to 75,000-150,000 hospitalisations and up to 600,000 medical encounters annually. Nevertheless, in 2022, only 18 countries in Europe (out of 38) have a publicly funded routine universal mass immunisation programme against rotavirus gastroenteritis. Evidence available in the last few years re-emphasises that rotavirus vaccines currently available in Europe are highly effective, preventing up to 96% of rotavirus-related hospitalisations in children less than 1 year of age (potentially 72,000-144,000 hospitalisations Europe-wide). Long-term surveillance indicates that rotavirus vaccination does not result in an overall increase in intussusception. On the contrary, increasing evidence suggests an overall reduction in intussusception in the first 12 months of life when early, high rotavirus vaccine coverage is achieved. Prevention of rotavirus gastroenteritis has marked positive impacts on parental wages and government tax revenue, with benefits extending across the whole economy. In the SARS-CoV-2 pandemic setting there is a new imperative to achieve high levels of paediatric vaccination against vaccine-preventable diseases, including rotavirus gastroenteritis. The introduction of rotavirus universal mass vaccination can be expected to reduce the number of preventable illnesses, hospitalisations and deaths caused by rotavirus gastroenteritis. Reducing vaccine-preventable diseases is particularly urgent at this time when healthcare systems are preoccupied and overwhelmed with SARS-CoV-2. Graphical abstract available for this article.