Bovine viral diarrhea virus Erns protein expressed in Chinese hamster ovary cells and its immunogenicity analysis / 生物工程学报

The aim of this study was to produce Erns protein of bovine viral diarrhea virus (BVDV) by using suspensively cultured CHO cells expression system and to analyze the immunogenicity of the purified Erns protein. In this study, the recombinant eukaryotic expression plasmid pcDNA3.1-BVDV-Erns was constructed based on the gene sequence of BVDV-1 NADL strain. The Erns protein was secreted and expressed in cells supernatant after transfecting the recombinant expression plasmid pcDNA3.1-BVDV-Erns into CHO cells. The expression and purification of the Erns protein was analyzed by SDS-PAGE, the reactivity was determined with anti-His monoclonal antibodies and BVDV positive serum with Western blotting. Immunogenicity analysis of the Erns protein was determined after immunizing New Zealand white rabbits, and the serum antibodies were tested by indirect ELISA (iELISA) and indirect immunofluorescence (IFA). The serum neutralizing titer of the immunized rabbits was determined by virus neutralization test. The concentration of the purified Erns protein was up to 0.886 mg/mL by BCA protein quantification kit. The results showed that the Erns protein could be detected with anti-His monoclonal antibodies and anti-BVDV sera. Serum antibodies could be detected by iELISA on the 7th day post-prime immunization, and the antibody level was maintained at a high titer until the 28th day post-immunization. The antibody titer was 1:128 000. Furthermore, the expression of the Erns protein in BVDV-infected MDBK cells could be detected with immunized rabbits sera by IFA. Moreover, antigen-specific neutralizing antibodies of 2.71 log10 was induced in rabbits. In this study, purified BVDV Erns protein was successfully produced using CHO suspension culture system, and the recombinant protein was proved to have a good immunogenicity, which may facilitate the development of BVD diagnosis method and novel subunit vaccine.

Surveillance results of viral diarrhea in children under five years of age in Shaoxing City / 预防医学

Objective@#To investigate the epidemiological and etiological characteristics of viral diarrhea among children under 5 years of age in Shaoxing City, Zhejiang Province, so as to provide insights into management of viral diarrhea.@*Methods@#The surveillance data on viral diarrhea among children under 5 years of age in Shaoxing City from 2019 to 2022 were collected, including demographics and stool testing results. The epidemiological and etiological characteristics of viral diarrhea were analyzed using a descriptive epidemiological method. @*Results@#A total of 763 diarrheal children under 5 years of age were reported in Shaoxing City from 2019 to 2022, and 236 children were tested positive for virus (30.93%). The detection of virus was 49.01%, 31.61%, 20.43% and 21.89% from 2019 to 2022, which appeared an overall tendency towards a decline (P<0.05). The incidence of viral diarrhea peaked from November to March of the next year and from May to July, and the detection of virus was lower among children with diarrhea living in urban areas (Yuecheng District, Keqiao District and Shangyu District) than among those in suburb areas (Zhuji City, Shengzhou City and Xinchang County) (30.22% vs. 52.00%, P<0.05). There were 206 children tested positive for a single virus, and the detection rates of rotavirus (RV), enteric adenovirus (EAdV), norovirus (NoV), and sapovirus (SaV) were 9.57%, 8.91%, 8.39%, and 0.13%, respectively. There were 25 children with virus co-infections, and the positive rates of EAdV and NoV, RV and EAdV and RV and NoV co-infections were 1.31%, 1.18% and 0.79%, respectively. There were 5 children with triple infections of RV, EAdV and NoV (0.66%). The highest detection of EAdV was seen in April, the highest detection of RV and NoV was seen in January, while SaV was only detected in April. @*Conclusion@#The incidence of viral diarrhea among children under 5 years of age peaked in winter and spring in Shaoxing City from 2019 to 2022, and the cases predominantly occurred in urban areas. The detection of virus appeared an overall tendency towards a decline, with high detection of RV, EAdV and NoV.

Pathogenic surveillance of virus diarrhea in Chongqing in 2018 - 2019 / 公共卫生与预防医学

Objective To understand the pathogen composition of viral diarrhea in Chongqing, and to provide reference for the prevention and control of viral diarrhea. Methods Real-time fluorescent RT-PCR was used to detect the nucleic acids of rotavirus, norovirus, sapovirus, astrovirus, and enteric adenovirus collected from diarrhea outpatient cases from 2018 to 2019, and the positive nucleic acid samples were sequenced. Results Among the 398 cases of diarrhea, 184 cases were detected positive, with the positive detection rate of 46.23%. Norovirus infection was the main infection, accounting for 29.40%. The G/P genotype of group A rotavirus was mainly G9P8, accounting for 90.32%. The genotype of norovirus was mainly GII.2[P16], accounting for 33.91%. The genotype of sapovirus was mainly GI.2, accounting for 55.56%. The genotype of astrovirus was HAstV-4, accounting for 100%. The genotype of enteric adenovirus was F41, accounting for 100%. The diarrhea cases were mainly distributed in the fourth quarter, with the positive detection rate of 70.42%, among which norovirus had the highest detection rate, accounting for 53.99%. Conclusion High incidence of viral diarrhea is in winter in Chongqing. The main pathogen of viral diarrhea is norovirus, and the genotypes of norovirus show diversity. It is necessary to prevent the outbreak and epidemic caused by norovirus in winter. In the future, the surveillance of viral diarrhea should be strengthened, and the viral diarrhea gene database should be improved to provide a scientific basis for epidemic prevention and control.

Molecular epidemiological characteristics of the virus in 96 children with acute diarrhea in Changdu of Tibet, China / 中国当代儿科杂志

OBJECTIVES@#To study the molecular epidemiological characteristics of the virus in children with acute viral diarrhea in Changdu of Tibet, China.@*METHODS@#Fecal specimens were collected from 96 children with acute diarrhea who visited the People's Hospital of Changdu, Tibet, from November 2018 to November 2020 and were tested for adenovirus, norovirus, astrovirus, sapovirus, and rotavirus. Gene sequencing was performed for the genotypes of these viruses.@*RESULTS@#The overall positive rate of the five viruses was 39% (37/96), among which astrovirus had the highest positive rate of 17%, followed by norovirus (9%), rotavirus (8%), adenovirus (7%), and sapovirus (5%). There was no significant difference in the positive rate of the five viruses among different age groups (P>0.05). Only the positive rate of astrovirus was significantly different among the four seasons (P<0.05). For adenovirus, 6 children had F41 type and 1 had C2 type; for norovirus, 6 had GⅠ.3 type, 1 had GⅠ.7 type, 1 had GⅡ.3 type, and 2 had GⅡ.4 Sydney_2012 type; HAstrV-1 type was observed in all children with astrovirus infection; for sapovirus, 1 child each had sporadic GⅠ.2, GⅠ.6, and GⅡ.1 sapovirus and 2 children had unknown type; 6 children had rotavirus G9[P8].@*CONCLUSIONS@#Astrovirus and norovirus are important pathogens in children with acute diarrhea in Changdu, Tibet. The positive rate of adenovirus, norovirus, astrovirus, sapovirus, and rotavirus is not associated with age, and only the positive rate of astrovirus has obvious seasonality. F41 type is the dominant genotype of adenovirus; GⅠ.3 is the dominant genotype of norovirus; HAstrV-1 is the dominant genotype of astrovirus; sporadic GⅠ.2, GⅠ.6, and GⅡ.1 are the dominant genotypes of sapovirus; G9[P8] is the dominant genotype of rotavirus.

Changes of common pathogens of viral diarrhea in children during the COVID-19 epidemic / 中华临床感染病杂志

Objective:To analyze the changes of common pathogens of viral diarrhea in children during the COVID-19 epidemic.Methods:A retrospective analysis was performed on children with acute diarrhea treated in Children’s Hospital affiliated to Zhejiang University School of Medicine from 2019 April to 2019 December(before COVID-19, n=407)and 2020 April to 2020 December (during COVID-19, n=645). Children were further divided into 1-6 months, >6 months-2 years, >2-5 years age groups. Detection rates of norovirus GI (NOVI), norovirus GⅡ (NOVⅡ), rotavirus A (ROVA), rotavirus C (ROVC), intestinal adenovirus (EADV), sapovirus (SAV) and astrovirus (ASV) in different age groups and seasons before and during COVID-19 were compared by χ2 or Fisher exact test. Results:The total detection rates of diarrhea-causing viruses among age groups were significantly higher before COVID-19 than those during COVID-19( χ2=8.43, 38.22 and 9.23, all P<0.05). The detection rates of NOVⅡ and EADV in infants aged 1-6 months and >6 months-2 years were decreased during the period of COVID-19 epidemic( χNOVⅡ2 =36.87 and 17.77, both P<0.001, χEADV2 =9.08, P=0.014 and 0.003); the detection rates of NOVⅡ in children aged 2-5 years was decreased during the period of COVID-19 epidemic( χ2=3.96, P=0.047); the detection rates of other diarrhea-causing viruses were not decreased during the period of COVID-19 epidemic(all P>0.05). The detection rates of diarrhea-causing viruses among 4-6 months, 7-9 months and 10-12 months were higher during the period of COVID-19 epidemic( χ2=11.62, 65.41 and 27.80, all P<0.001). Conclusion:After the outbreak of COVID-19, the detection rate of common pathogens of viral diarrhea in children is decreased, especially NOVⅡ and EADV, which may be related to the measures in response to major public health events in Zhejiang Province.

Pathogenic surveillance of viral diarrhea in Minhang District of Shanghai in 2014-2020 / 公共卫生与预防医学

Objective To investigate the epidemiological and pathogenic characteristics of viral diarrhea in Minhang District, Shanghai. Methods Random sampling on diarrhea was conducted in intestinal outpatient departments of 2 sentinel hospitals according to a certain sampling interval in Minhang District，Shanghai from 2014 to 2020. Real time PCR technology was used to detect Rotavirus, Norovirus, adenovirus, Astrovirus and Sapovirus in fecal samples. Results A total of 646 out of 1 839 stool specimenswere tested positive, and the positive rate was 35.13%.Five pathogenic viruses were detected , mostly norovirus (421 cases, 65.17%) followed by rotavirus (151 cases, 23.37%).The positive rate of norovirus was higher in the age group of 20- 69 years, and the positive rate of Rotavirus was higher in the age group of 0- 9 years. Conclusions Norovirus and rotavirus accounted for the majority of reported infection diarrhea cases in MinhangDistrict of Shanghai from 2014 to 2020, with significant seasonal peaks. Tailored prevention and control measures should be carried out, particularly in risk seasons.

Preparation of bovine viral diarrhea disease virus 1 virus-like particles and evaluation of its immunogenicity in a guinea pig model / 生物工程学报

In order to obtain virus-like particles (VLPs) for prevention of bovine viral diarrhea virus 1 (BVDV-1), the C-Erns-E1-E2 region was cloned into a pFastBacDaul vector for generating the recombinant Bacmid-BVDV-1 in DH10Bac Escherichia coli. The recombinant baculovirus Baculo-BVDV-1 was produced by transfecting the Sf9 cells with Bacmid-BVDV-1. The expressed protein and the assembled VLPs were determined by immunofluorescence, Western blotting and electron microscopy. Guinea pigs were immunized with inactivated VLPs coupled with the Montanide ISA-201 adjuvant. The immunogenicity of VLPs was evaluated by monitoring the humoral immune response with neutralizing antibody titer determination, as well as by analyzing the cell-mediated immune response with lymphocyte proliferation assay. The protective efficacy of VLPs was evaluated by challenging with 106 TCID50 virulent BVDV-1 strain AV69. The results showed that the recombinant Baculo-BVDV-1 efficiently expressed BVDV structural protein and form VLPs in infected Sf9 cells. The immunization of guinea pigs with VLPs resulted in a high titer (1:144) of neutralizing antibody, indicating an activated cellular immunity. Significantly lower viral RNA in the blood of the post-challenged immunized guinea pigs was observed. The successful preparation of BVDV VLPs with insect cell expression system and the observation of the associated immunogenicity may facilitate further development of a VLPs-based vaccine against BVD.

Analysis of pathogen detection of viral diarrhea in Southwest China from 2017 to 2019 / 公共卫生与预防医学

Objective To understand the dynamic changes of the pathogen spectrum of viral diarrhea in Southwest China, and to provide a scientific basis for formulating prevention and control measures. Methods A total of 2 262 fecal samples of diarrhea cases were collected from the diarrhea monitoring sites in Yunnan, Sichuan, and Chongqing from January 2017 to December 2019. The detection of rotavirus, norovirus, adenovirus, astrovirus, and sapovirus was performed by real-time PCR, and their epidemiological data were collected. Results The ratio of male to female patients was 1.30:1. The majority of patients were 3 years old and younger, accounting for 52.21%% of all cases. Children living at home accounted for 49.38% of all cases. The detection rate of rotavirus was 10.70%, followed by norovirus at 10.43% and adenovirus at 4.60%. The detection rate of rotavirus was the highest in Yunnan and Chongqing, which were 20.18% and 16.96%, respectively, while the detection rate of norovirus was the highest in Sichuan, which was 13.54%. Conclusion Rotavirus was still the main diarrheal pathogen in Southwest China. High incidence of diarrhea occurred in autumn and winter, with children living at home being the main patients. Measures should be taken to prevent outbreaks caused by norovirus and adenovirus.

Prevalence of bovines persistently infected with bovine viral diarrhea virus (BVDV) in dairy cattle herds in Paraná State, Brazil

ABSTRACT: This study aimed to establish the prevalence of animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) in dairy farms at Parana State, Brazil. Samples were collected from 6,465 female Holstein Friesian Dairy Cattle, including animals less than two years old, females over two years old who had not given birth at the farm, and mothers of calves diagnosed as persistently infected. The cattle came from 40 dairy herds distributed in 10 municipalities in the State of Paraná. The samples were obtained from May 2015 to August 2018. The diagnosis of PI animals was made with an antigen-capture ELISA test. We detected PI animals in fifteen herds sampled (37.5%), ranging from one to sixteen animals per herd. The prevalence in Parana States municipalities was 1.78%, ranging from 0.3 to 8.9% at positive herds. The analysis of the individual herds shows significant dissemination of the BVDV in Paranas municipalities, including endemic areas. With this, we highlight the need for measures to raise awareness among producers about the existence and importance of bovine viral diarrhea (BVD) in dairy herds, reinforcing the PI animals role in disease epidemiology and the economic impact caused by the maintenance of these farm animals.

RESUMO: Com o intuito de se estabelecer a prevalência de animais persistentemente infectados (PI) com o BVDV em propriedades leiteiras no estado do Paraná. Foram coletadas amostras de 6.465 bovinos, fêmeas, da raça Holandês Preto e Branco (HPB). Amostraram-se animais com idade inferior a dois anos, fêmeas com mais de dois anos que não haviam tido partos na propriedade, e mães de bezerros que foram diagnosticados como persistentemente infectados. Os bovinos foram provenientes de 40 rebanhos leiteiros, distribuídos em 10 municípios no Estado do Paraná. A coleta deu-se no período de maio de 2015 a agosto de 2018. O diagnóstico dos animais PI foi feito por meio do teste de ELISA de captura de antígeno. Animais PI foram detectados em quinze rebanhos amostrais (37,5%), oscilando entre um e dezesseis animais por rebanho. A prevalência nos municípios do estado Paraná foi de 1,78%, oscilando entre 0,3 a 8,9% nos rebanhos positivos. Com a alta prevalência de animais PI observada, quando analisados os rebanhos amostrais individualmente, é possível afirmar que há uma disseminação importante do BVDV em municípios paranaenses, destacando inclusive áreas endêmicas. Com isso, vê-se a necessidade de medidas de conscientização dos produtores sobre a existência e importância da BVD nos rebanhos, destacando o papel dos animais PI na epidemiologia da doença, bem como o impacto econômico causado pela manutenção desses animais nos rebanhos.

Prevalence of bovines persistently infected with bovine viral diarrhea virus (BVDV) in dairy cattle herds in Paraná State, Brazil / Prevalência de bovinos persistentemente infectados com o vírus da diarreia viral bovina (BVDV) em rebanhos leiteiros no estado do Paraná, Brasil

This study aimed to establish the prevalence of animals persistently infected (PI) with bovine viral diarrhea virus (BVDV) in dairy farms at Parana State, Brazil. Samples were collected from 6,465 female Holstein Friesian Dairy Cattle, including animals less than two years old, females over two years old who had not given birth at the farm, and mothers of calves diagnosed as persistently infected. The cattle came from 40 dairy herds distributed in 10 municipalities in the State of Paraná. The samples were obtained from May 2015 to August 2018. The diagnosis of PI animals was made with an antigen-capture ELISA test. We detected PI animals in fifteen herds sampled (37.5%), ranging from one to sixteen animals per herd. The prevalence in Parana State's municipalities was 1.78%, ranging from 0.3 to 8.9% at positive herds. The analysis of the individual herds shows significant dissemination of the BVDV in Parana's municipalities, including endemic areas. With this, we highlight the need for measures to raise awareness among producers about the existence and importance of bovine viral diarrhea (BVD) in dairy herds, reinforcing the PI animals' role in disease epidemiology and the economic impact caused by the maintenance of these farm animals.(AU)

Com o intuito de se estabelecer a prevalência de animais persistentemente infectados (PI) com o BVDV em propriedades leiteiras no estado do Paraná. Foram coletadas amostras de 6.465 bovinos, fêmeas, da raça Holandês Preto e Branco (HPB). Amostraram-se animais com idade inferior a dois anos, fêmeas com mais de dois anos que não haviam tido partos na propriedade, e mães de bezerros que foram diagnosticados como persistentemente infectados. Os bovinos foram provenientes de 40 rebanhos leiteiros, distribuídos em 10 municípios no Estado do Paraná. A coleta deu-se no período de maio de 2015 a agosto de 2018. O diagnóstico dos animais PI foi feito por meio do teste de ELISA de captura de antígeno. Animais PI foram detectados em quinze rebanhos amostrais (37,5%), oscilando entre um e dezesseis animais por rebanho. A prevalência nos municípios do estado Paraná foi de 1,78%, oscilando entre 0,3 a 8,9% nos rebanhos positivos. Com a alta prevalência de animais PI observada, quando analisados os rebanhos amostrais individualmente, é possível afirmar que há uma disseminação importante do BVDV em municípios paranaenses, destacando inclusive áreas endêmicas. Com isso, vê-se a necessidade de medidas de conscientização dos produtores sobre a existência e importância da BVD nos rebanhos, destacando o papel dos animais PI na epidemiologia da doença, bem como o impacto econômico causado pela manutenção desses animais nos rebanhos.(AU)

Co-infection by Neopora caninum and bovine viral diarrhea virus in cattle from Rio Grande do Sul, Brazil, destined to exportation / Coinfecção por Neospora caninum e vírus da diarreia viral bovina em bovinos do Rio Grande do Sul, Brasil, destinados à exportação

Reproductive tests in cattle are of great economic importance, given the impact it can have on the production system and may be caused by agents. Neospora caninum and Bovine Viral Diarrhea virus (BVDV) are considered of great importance as reproductive and should be considered responsible for keeping animals persistently infected. The present study included 479 calf serum samples for export in the state of Rio Grande do Sul (RS). All samples were screened for BVDV by an ELISA antigen. BVDV antigen-positive ELISA samples were isolated from BVDV in cell culture. An indirect immunofluorescence (IFT) technique was used to detect anti-N. caninum antibodies. Of the 479 export-treated serum samples, 361 were positive for BVDV antigens by ELISA and/or viral isolation test (361/479-75.36%), and 109 IFT-positive samples for N. caninum (109/479-22.75%). Despite detection of antibodies anti-N. caninum did not differ statistically between naturally infected BVDV and non-BVDV infected animals suggesting that there is no interference of BVDV infection on infection or detection rate of animals with N. caninum, positive animals in viral isolation and high DO in BVDV-Ag ELISA. may present active disease and consequent immunosuppression, contributing to a potential reactivation of N. caninum.(AU)

Testes reprodutivos em bovinos são de grande importância econômica, dado o impacto que podem ter no sistema de produção e podem ser causados por agentes. O Neospora caninum e o vírus da Diarreia Viral Bovina (BVDV) são considerados de grande importância como reprodutivos e devem ser considerados responsáveis por manter os animais persistentemente infectados. O presente estudo incluiu 479 amostras de soro de bezerro para exportação no estado do Rio Grande do Sul (RS). Todas as amostras foram rastreadas para BVDV por um antígeno ELISA. As amostras de ELISA positivas para o antigénio BVDV foram isoladas a partir de BVDV em cultura de células. Uma técnica de imunofluorescência indireta (IFT) foi utilizada para detectar anticorpos anti-N caninum. Das 479 amostras de soro tratadas para exportação, 361 foram positivas para antígenos de BVDV por ELISA e/ou teste de isolamento viral (361/479-75,36%) e 109 amostras positivas para IFT para N. caninum (109/479-22,75%). Apesar da detecção de anticorpos anti-N. caninum não diferiu estatisticamente entre animais infectados naturalmente BVDV e não BVDV sugerindo que não há interferência da infecção pelo BVDV na infecção ou taxa de detecção de animais com N. caninum, animais positivos em isolamento viral e alta DO em BVDV-Ag ELISA, pode apresentar doença ativa e consequente imunossupressão, contribuindo para uma potencial reativação de N. caninum.(AU)

Pestivirus spillover effect: molecular detection of bovine viral diarrhea virus in domestic and feral pigs / Efeito spillover de pestivírus: detecção molecular do vírus da diarreia viral bovina em suínos domésticos e javalis

Pestivirus infections are important in the livestock industries, with infection occurring in cattle, sheep and pigs. The Pestivirus genus of the family Flaviviridae, includes four recognized species: bovine viral diarrhea virus 1 (BVDV-1), bovine viral diarrhea virus 2 (BVDV-2), border disease virus (BDV), and classical swine fever virus (CSFV). All pestivirus species can infect pigs, therefore accurate and specific pestivirus detection and differentiation is of great importance to assure control measures in swine populations. The aim of the study was the molecular detection of different pestiviruses in domestic and feral pigs. A total of 527 samples (92 pigs and 435 wild boars) were tested for pestiviruses detection using molecular assays. Eleven positive samples (6 wild boars and 5 domestic pigs) were identified using panpestivirus primers targeting the 5'- UTR region of the pestivirus RNA genome. Further all the positive samples were sequentially tested for detection of CSFV, BVDV-1 and BVDV-2 using specific primers. All RNAs were identified as positives for BVDV-1 and no amplification signals were obtained from BVDV-2 and CSFV. The current detection of BVDV-1 in clinical swine specimens highlights the important risk factor of swine population as reservoir and consequently carrier for BVDV.(AU)

As infecções por pestivírus são importantes nas indústrias pecuárias, com infecções em bovinos, ovinos e suínos. O gênero Pestivirus da família Flaviviridae inclui quatro espécies reconhecidas: vírus da diarreia viral bovina 1 (BVDV-1), vírus da diarreia viral bovina 2 (BVDV-2), vírus da doença de fronteira (VDF) e vírus da peste suína clássica (VPSC). Todas as espécies de pestivírus podem infectar porcos, portanto a detecção e diferenciação precisas e específicas de pestivírus são de grande importância para garantir medidas de controle nas populações suínas. O objetivo do estudo foi a detecção molecular de diferentes pestivírus em suínos domésticos e javali. Um total de 527 amostras (92 porcos e 435 javalis) foram testados para detecção de pestivírus usando ensaios moleculares. Onze amostras positivas (6 javalis e 5 porcos domésticos) foram identificadas usando iniciadores de panpestivírus visando a região 5'-UTR do genoma do RNA do pestivírus. Além disso, todas as amostras positivas foram testadas sequencialmente para detecção de VPSC, BVDV-1 e BVDV-2 usando iniciadores específicos. Todos os RNAs foram identificados como positivos para BVDV-1 e nenhum sinal de amplificação foi obtido do BVDV-2 e CSFV. A detecção atual do BVDV-1 em amostras clínicas de suínos destaca o importante fator de risco da população suína como reservatório e consequentemente portador do BVDV.(AU)

Investigation on the epidemiological characteristics of viral diarrhea among children under 5 years in Beijing Xicheng District, 2015-2018 / 公共卫生与预防医学

Objective To investigate the epidemiological characteristics of viral diarrhea in children under 5 years old during 2015-2018 in Xicheng District of Beijing, and to provide evidence for prevention and control of viral diarrhea. Methods Fecal specimens from patients under 5 years old were collected from the outpatient clinic of the Children's Diarrhea Surveillance Sentinel Hospital in Xicheng District, Beijing from 2015 to 2018. Real-time fluorescence PCR was used to detect group A rotavirus (RVA), norovirus (NV), and adenovirus (AdV), Astrovirus (AstV). Results A total of 972 fecal specimens were collected, 142 were tested positive for RVA (14.61%), 136 were tested positive for NV (13.99%), 67 were tested positive for AdV (6.89%), and 49 were tested positive for AstV (5.04%). Mix-infection was found in 46 cases, which accounted for 4.73%. The detection rates of the 4 viruses in children under 2 years old were 80.28%, 77.94%, 83.58%, and 87.75%, respectively. Conclusion Rotavirus and norovirus were the main pathogens of viral diarrhea in children in Beijing Xicheng District from 2015-2018, and iinfants under 2 years old were the high-risk group. The peak incidence was in winter.

Serological response against bovine herpesvirus and bovine viral diarrhea virus induced by commercial vaccines in Holstein heifers / Resposta sorológica contra herpesvírus bovino e vírus da diarreia viral bovina induzida por vacinas comerciais em novilhas Holandesas

Vaccination is a strategy to the prevention and control of reproductive diseases caused by bovine viral diarrhea virus (BVDV) and bovine herpesvirus type 1 (BoHV-1), however the various compositions of commercial vaccines should be evaluated for their ability to induce protection mediated by antibodies. The objective of this research was to evaluate the production of specific neutralizing Abs against BVDV-1 and 2, and BoHV-1 induced by commercial vaccines composed by different adjuvants. Holstein heifers were vaccinated and distributed in three experimental groups: Group I (G1) was vaccinated with a commercial vaccine containing inactivated BVDV-1, BVDV-2 and BoHV-1 diluted in alum hydroxide as adjuvant (n=9); Group II (G2) was vaccinated with an product containing inactivated strains of BVDV-1, BVDV-2, BoHV-1 and BoHV-5 diluted in oil emulsion as adjuvant (n=10); Group III (G3) was vaccinated with a commercial vaccine containing inactivated BVDV-1 and BVDV-2, besides live modified thermosensitive BoHV-1, diluted in Quil A, amphigen and cholesterol (n=10); A control, non-vaccinated group (n=6) was mock vaccinated with saline. Heifers received two subcutaneous doses of 5mL of each commercial vaccine on the right side of the neck, with 21 days interval. Humoral immune response was assessed by the virus neutralization test (VN) against BVDV-1 (NADL and Singer strains), BVDV-2 (SV253 strain) and BoHV-1 (Los Angeles strain) in serum samples collected on vaccination days zero (D0), 21 (D21) and 42 (D42; 21 days after boosting). Neutralizing Abs against BVDV-1 NADL was detected only in D42, regardless of the vaccine used. Similar geometric mean titers (GMT) for BVDV-1 NADL were observed between G1 (log2=5.1) and G3 (log2=5.1). The seroconversion rate (%) was higher in G1 (78%) when compared to G2 (10%) and G3 (40%). For BVDV-1 Singer, it was also possible to detect Abs production in G1 (log2=5.8, 100% seroconversion rate) and G3 (log2=3.5, seroconversion rate = 60%), only after the booster dose (D42). Neutralizing Abs to BVDV-2 (SV253) were detected only in G3, observing 90% seroconversion associated with high titers of Abs (log2=6.7) after the 2nd dose of vaccine (D42). Heifers from G1 and G3 responded to BoHV-1 after the first dose (D21): G1 (log2=2.5, seroconversion rate = 67%) and G3 (log2=0.7, seroconversion rate = 80%). In D42, a higher magnitude response was observed in the heifers from G3 (log2=6.1, 100%) compared with G1 (log2=4.3, 100%) and G2 (log2=2.7, 60%). Based on the data obtained, it can be concluded that the commercial vaccine contained aluminum hydroxide (G1) was most effective in the induction of antibodies against BVDV-1. On the other hand, this vaccine did not induce the production of neutralizing Abs against BVDV-2. Only the heifers from G3 (Quil A, amphigen and cholesterol) generated neutralizing Abs against BVDV-2. The animals that received commercial vaccine containing oil emulsion as adjuvant (G2) had a weak/undetectable response against BVDV-1 and BVDV-2. The best protective response against BoHV-1 was observed in heifers vaccinated with the live modified thermosensitive virus.(AU)

A vacinação é utilizada como estratégia para a prevenção e controle das doenças reprodutivas, causadas pelos vírus da diarreia viral bovina (BVDV) e herpesvírus bovino tipo 1 (BoHV-1), entretanto, as diversas composições de vacinas comerciais devem ser avaliadas quanto a sua eficiência protetiva mediada por anticorpos (Acs). O objetivo desta pesquisa foi avaliar a produção Acs neutralizantes específicos para cepas de BVDV-1 e 2, e BoHV-1 induzida por vacinas comerciais contendo diferentes tipos de adjuvantes. Para tal, novilhas Holandesas foram vacinadas e distribuídas em três grupos experimentais: Grupo I (G1) foi vacinado com uma vacina comercial composta por cepas inativadas de BVDV-1, BVDV-2 e BoHV-1 diluídas em hidróxido de alumínio como adjuvante (n=9); Grupo II (G2) foi vacinado com produto contendo as cepas inativadas de BVDV-1, BVDV-2, BoHV-1 e BoHV-5 em uma emulsão oleosa como adjuvante (n=10); O Grupo III (G3) foi vacinado com uma vacina comercial contendo BVDV-1 e BVDV-2 inativado, além do BoHV-1 vivo modificado e termosensivel, diluídos em adjuvante contendo Quil A, Amphigem e colesterol (n=10); O Grupo Controle não vacinado (n=6) foi inoculado com solução salina. As novilhas receberam duas doses das respectivas vacinas ou solução salina (5mL), com intervalo de 21 dias, por via subcutânea, na tábua do pescoço do lado direito. A resposta imune humoral foi avaliada pelo teste de vírus neutralização (VN) contra o BVDV-1 (cepas NADL e Singer), BVDV-2 (cepa SV253) e BoHV-1 (cepa Los Angeles) em amostras de soro coletadas nos dias (D) de vacinação zero (D0), 21 dias após 1ª dose (D21)e 42 (D42; 21 dias após A 2ª dose). Os anticorpos neutralizantes contra o BVDV-1 NADL foram detectados apenas em D42, independentemente da vacina utilizada. Os títulos médios geométricos (GMT) de anticorpos foram semelhantes entre G1 (log2=5,1) e G3 (log2=5,1). A taxa de soroconversão foi maior no G1 (78%) quando comparado ao G2 (10%) e G3 (40%). Para o BVDV-1 Singer, somente após D42 foi observada a produção de Acs no G1 (log2=5,8; taxa de soroconversão de 100%) e G3 (log2=3,5; taxa de soroconversão = 60%). Os anticorpos contra BVDV-2 (SV253) foram detectados apenas nas novilhas do G3, observando-se taxa de soroconversão de 90% com altos títulos de anticorpos neutralizantes (log2=6,7) em D42. Novilhas G1 e G3 responderam ao BoHV-1 após a primeira dose (D21): G1 (log2=2,5; taxa de seroconversão = 67%) e G3 (log2=0,7; taxa de seroconversão = 80%). Em contrapartida, foi observada uma maior magnitude de resposta para as novilhas G3 (log2=6,1; 100%) em D42, em relação aos animais G1 (log2=4,3; 100%) e G2 (log2=2,7; 60%). Com base nos dados obtidos, foi possível concluir que a vacina composta por hidróxido de alumínio (G1) foi mais eficaz na produção de anticorpos contra o BVDV-1, em contrapartida esse produto não induziu anticorpos contra o BVDV-2. Apenas as novilhas do G3 (Quil A, amphigen e colesterol) geraram Acs neutralizantes contra o BVDV-2. Os animais que receberam a vacina em emulsão oleosa (G2) como adjuvante apresentaram uma resposta fraca/indetectável contra o BVDV-1 e BVDV-2. A melhor resposta protetiva contra o BoHV-1 foi observada nas novilhas vacinadas com a vacina viva modificada termosensível.(AU)

Molecular epidemiology of norovirus among infants with diarrhea in Chaoyang district, Beijing from 2011 to 2017 / 中华实验和临床病毒学杂志

Objective@#To investigate the genetic characteristics of human norovirus (NoV) among infants under 5 years of age with diarrhea in Chaoyang District, Beijing from 2011 to 2017.@*Methods@#NoV-positive stool samples were collected from 2011 to 2017 in this region. The partial RdRp and VP1 genes were amplified and sequenced. Multi-sequence alignment was performed and phylogenetic tree was constructed using Mega software.@*Results@#A total of 151 samples were sequenced and analyzed. The ratio of male and female was 2.28∶1 with mean age of 1.72 years. Fourteen NoV subtypes were detected, including GII.Pe/GII.4 (47.68%), GII.P12/GII.3 (20.53%), GII.P4/GII.4 (17.22%), GII.P16/GII.2 (3.31%), GII.P12/GII.12 (1.99%), GII.P17/GII.17 (1.99%), GII.P16/GII.13 (1.32%), GII.P7/GII.7 (1.32%), GII.P7/GII.6 (1.32%), GII.P2/GII.2 (0.66%), GII.P21/GII.21 (0.66%), GII.Pg/GII.12 (0.66%), GI.Pa/GI.3 (0.66%) and GI.P6/GI.6 (0.66%).@*Conclusions@#NoV genetic diversity was found among infants under 5 with diarrhea in Chaoyang district, Beijing. The subtypes from surveillance and those from epidemics occurred in chronological order. The surveillance should be strengthened for early detection of new subtype for monitoring the epidemic and vaccine design.

Analysis of surveillance results of viral diarrhea among children under 5 years of age in Shandong province from 2012 to 2017 / 中华实验和临床病毒学杂志

Objective@#To analyze the epidemiological characteristics and pathogen types of viral diarrhea in children under 5 years of age in Shandong province, and provide reference for the prevention and control of viral diarrhea.@*Methods@#A total of 1 017 fecal samples were collected from all children aged 5 years and younger with diarrhea who were admitted to the sentinel hospital of Shandong province from 2012 to 2017 within 3 days. Rotavirus antigen was detected by using an ELISA method . Rotavirus G/P typing was performed by RT-PCR; Norovirus (GI and GII), Sapovirus, and Astrovirus were detected by multiplex RT-PCR, and adenovirus was detected by PCR.@*Results@#In the 1 017 fecal specimens, the overall positive rate was 51.62% (525/1017), and viral nucleic acids were detected in at least 421 samples, and mixed virus infection was found in 104 sampes. The mixed infection accounted for 10.23% (104/1017) of all infections. The positive detection rates of Rotavirus, Calicivirus, Adenovirus and Astrovirus were 34.22% (348/1017), 16.91% (172/1017), 2.56% (26/1017), and 9.64% (98/1017)). The total detection rate of diarrhea virus and the detection rate of Rotavirus were the highest at 12 to 17 months of age, which was 51.72% (105/203) and 20.20% (41/203), respectively. The positive rate of diarrhea in children aged 2 years and younger was 49.36% (502/1017), which was much higher than the positive rate of diarrhea in children over 2 years old (2.26% (23/1017)). The peak of viral diarrhea was found to occur between November and April of the following year. The genotype of rotavirus was dominated by G9 (82.76%), the P genotype was dominated by P[8] (80.46%), and the G/P combination was dominated by G9P[8] (83.87%). Norovirus was the main infection in the Calicivirus (87.21%).@*Conclusions@#From 2012 to 2017, viral diarrhea in children under 5 years of age in Shandong Province was mainly caused by Rotavirus infection, followed by Norovirus. The overall prevalence of viral diarrhea in Shandong was moderate in China, and autumn and winter were the main epidemic season for viral diarrhea.

Treating Viral Diarrhea in Children by Probiotic and Zinc Supplements / 대한소아소화기영양학회지

PURPOSE: The aim of this study was to analyze the effects of probiotics and zinc supplements on the mean duration and frequency of acute diarrhea in children aged 6 months to 2 years. METHODS: In this clinical trial of infants aged between 6 months and 2 years, eligible patients were divided into 3 groups: Zinc Receiving Group (ZRG), Probiotic Receiving Group (PRG), and a control group receiving supportive care alone. The frequency of diarrhea was evaluated in the test groups during the first 24 hours and 48–72 hours, along with the duration of hospitalization and diarrhea persistence for 3–7 days. RESULTS: Diarrhea persisted for until the third day of admission in 100% of the infants in PRG compared with only 76.1% in ZRG. The relative risk of diarrhea persistence in the PRG was 1.31 times more than in ZRG until the third day. Also, 80% of diarrhea cases in the PRG persisted until the fourth day of admission, compared with 47.8% in the ZRG group, and this value was significant. The relative incidence of diarrhea persistence in the PRG was 36.4 times greater than in the ZRG until the day 4. Also, the percentage of post-treatment complications was 35.5% in the PRG and 2.6% in the ZRG, which was significant. CONCLUSION: In our study, the effectiveness of zinc at a dose of 20 mg was higher than that of probiotics. The complications associated with zinc supplementation were lower than those of probiotics.

Prostaglandin A1 inhibits the replication of bovine viral diarrhea virus

ABSTRACT Bovine viral diarrhea virus can cause acute disease in livestock, leading to economic losses. We show that Prostaglandin A1 inhibits bovine viral diarrhea virus replication in Madin-Darby bovine kidney cells (94% inhibition using 5 µg/mL). Light and electron microscopy of infected cells shows that Prostaglandin A1 also prevents virus-induced vacuolization, but at higher concentrations (10 µg/mL).

Evidence of bovine viral diarrhea virus transmission by back pond water in experimentally infected piglets / Evidência da transmissão do vírus da diarreia viral bovina através da lâmina d'água em leitões experimentalmente infectados

Swine can be infected by bovine viral diarrhea virus (BVDV). However, transmission routes among pigs are still unknown. The objective of the present study was to induce experimental infection of BVDV-1 in weaned piglets and to assess the potential transmission through pen back pond water, used to facilitate heat exchange of the pigs housed in barns. Two repetitions (BP1 and BP 2) were performed using 12 piglets proven to be free BVDV (n=6 per repetition) allocated into three groups: control, sentinels and infected with two piglets each. The piglets were placed in stainless steel isolators. The infected group received an inoculum containing BVDV-1, Singer strain. The piglets remained in the cabinets for 25 days, during which samples of nasal swab were collected daily and blood sampled weekly. At the end, the piglets were euthanized, necropsied and organ fragments were collected for histopathology, immunohistochemistry and RT-PCR. In the first experiment (BP1) the infected animals shed the virus between days 6 and 21 post-infection. Regarding the sentinel group, shedding occurred in only one piglet, on the 20th day after infection, and seroconversion was observed on the 25th day post-infection. In BP2, infected piglets I3 and I4 shed the virus on days 4 and 21 post-infection, respectively. Only one sentinel piglet (S3) she the virus on day 13 post-infection. Therefore, it was concluded that pigs can become infected with BVDV-1 and shed potentially infectious viral particles consequently, being able to transmit the virus to other pigs through back pond water.(AU)

Os suínos podem ser infectados pelo vírus da diarreia viral bovina (BVDV). No entanto, as vias de transmissão entre os suínos são ainda desconhecidas. O objetivo do presente estudo foi induzir a infecção experimental de BVDV-1 em leitões desmamados e avaliar a potencial transmissão pela lâmina d'água, que ajuda na troca de calor dos suínos alojados em baias. Duas repetições do experimento (BP1 e BP2) foram realizadas com 12 animais comprovadamente livres de BVDV (n=6 por repetição) alocados em três grupos: controle, sentinelas e infectados, com dois animais cada. Os animais foram mantidos em isoladores de aço inoxidável. O grupo infectado recebeu um inóculo contendo BVDV-1, estirpe Singer. Os animais permaneceram nos isoladores durante 25 dias e, durante esse período, amostras de suabe nasal foram coletadas diariamente e sangue coletado semanalmente. No final, os animais foram eutanasiados, necropsiados e fragmentos de órgãos foram coletados para histopatologia, imuno-histoquímica e RT-PCR. No primeiro experimento (BP1), os animais infectados excretaram partículas virais entre os dias 6 e 21 pós-infecção. Quanto ao grupo sentinela, a excreção ocorreu apenas em um animal, no 20º dia pós-infecção, e a soroconversão foi observada no 25º dia pós-infecção. Na BP2, os animais infectados I3 e I4 excretaram partículas virais nos dias 4 e 21 pós-infecção, respectivamente. Apenas um animal sentinela (S3) apresentou excreção no dia 13 pós-infecção. Concluiu-se que os suínos podem se infectar com BVDV-1 e excretar partículas virais potencialmente infecciosas, sendo capazes de transmitir o vírus a outros suínos através da lâmina d'água.(AU)

Identificación de factores asociados con la exposición al virus de la diarrea viral bovina (VDVB) en terneras de hatos lecheros de la sabana de Bogotá / Identification of factors associated with exposure to bovine viral diarrhea virus (BVDV) in dairy-herd calves in the Bogotá savanna / Identificação de fatores associados com a exposição ao vírus da diarreia viral bovina (VDVB) em bezerras de fazendas de gado leiteiro da savana de Bogotá

Resumen El virus de la diarrea viral bovina (VDVB) es un patógeno que afecta la salud bovina; produce signos clínicos como bronconeumonía, diarrea, teratogenia y pérdidas reproductivas. El objetivo del estudio fue determinar la seroprevalencia y algunos factores asociados con la exposición al VDVB en un total de 930 terneras provenientes de 31 hatos lecheros de la sabana de Bogotá. Los datos de los hatos se obtuvieron a través de una encuesta epidemiológica, así como la información individual de las terneras. Se utilizó una prueba de ELISA para la detección de anticuerpos contra VDVB p80. Los datos se analizaron con estadística de frecuencias, descriptiva, análisis univariado y con un modelo de regresión logística binaria en el que se evaluaron factores de confusión a través de la comparación entre los odds ratio (OR) crudos y ajustados. La seroprevalencia promedio de VDVB fue del 27,1 % (rango 0-90 %); adicionalmente, en el 83,9 % de los hatos se observaron anticuerpos contra VDVB. Los factores asociados con la exposición a VDVB fueron: 1) la edad en animales menores de 4 meses (OR = 4,9; IC 95 %: 2,52-9,56), 2) histórico de aborto de la madre (OR = 6,4; IC 95 %: 3,91-10,46; p = 0,001) y 3) la presentación histórica de diarrea (OR = 2,6; IC 95 %: 1,58-4,46; p = 0,010). Este estudio permitió confirmar la alta exposición al virus en los hatos e identificar de algunos factores asociados, lo cual contribuye al conocimiento de la epidemiología de la enfermedad.

Abstract Bovine viral diarrhea virus (BVDV) is a pathogen that affects bovine health; it produces clinical signs such as bronchopneumonia, diarrhea, teratogenicity, and reproductive losses. This study aimed to determine seroprevalence and some other factors associated with exposure to BVDV in a total of 930 calves from 31 dairy herds in the Bogotá savanna. The data of the herds, as well as individual information of the calves, were collected through an epidemiological survey. An ELISA test was used to detect antibodies to BVDV p80. Data were analyzed with frequency and descriptive statistics, univariate analysis, and a binary logistic regression model, in which confounding factors were evaluated through a comparison between crude and adjusted odds ratios (OR). The average seroprevalence of BVDV was 27.1% (range 0-90%); additionally, antibodies against BVDV were observed in 83.9% of the herds. The factors associated with exposure to BVDV were: 1) age of animals under 4 months (OR = 4.9; 95% CI: 2.52-9.56); 2) records of abortion of the mother (OR = 6.4; 95% CI: 3.91-10.46; p = 0.001); and 3) records of diarrhea (OR = 2.6; 95% CI: 1.58-4.46; p = 0.010). This study allowed confirming a high exposure to the virus in these herds and identifying some associated factors, which contributes to a better understanding of the epidemiology of the disease.

Resumo O vírus da diarreia viral bovina (VDVB) é um patógeno que afeta a saúde bovina; produz sinais clínicos como broncopneumonia, diarreia, teratógena e perdas reprodutivas. O objetivo do estudo foi determinar a soro prevalência e alguns fatores associados com a exposição ao VDVB em um total de 930 bezerras provenientes de 31 fazendas de gado leiteiro da savana de Bogotá. Os dados dos rebanhos foram obtidos através de uma enquete epidemiológica, assim como a informação individual das bezerras. Utilizou-se uma prova de ELISA para detecção de anticorpos contra VDVB p80. Os dados foram analisados com estatística de frequências, descritiva, análise uni variada e com um modelo de regressão logística binária no qual foram avaliados fatores de confusão através da comparação entre os odds ratio (OR) crus e ajustados. A soro prevalência média de VDVB foi do 27,1 % (rango 0-90 %); adicionalmente, no 83,9 % dos rebanhos se observaram anticorpos contra VDVB. Os fatores associados com a exposição a VDVB foram: 1) a idade em animais menores de 4 meses (OR = 4,9; IC 95 %: 2,52-9,56), 2) histórico de aborto da madre (OR = 6,4; IC 95 %: 3,91-10,46; p = 0,001) e 3) a apresentação histórica de diarreia (OR = 2,6; IC 95 %: 1,58-4,46; p = 0,010). Este estudo permitiu confirmar a alta exposição ao vírus nos rebanhos bovinos e identificar de alguns fatores associados, fator que contribui para o conhecimento da epidemiologia da doença.