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In Vietnam, diarrhea, especially persistent diarrhea, is one of the most common diseases in children, while a significant proportion of cases are negative with pathogens; thus, there is an urgent need to understand gut bacterial dysbiosis. In this study, bacteria in the fecal samples of five healthy and ten diarrheal children were separated from other residues, then adopted to extract their metagenomic DNA for evaluating their diversity based on V3 and V6–V8 regions and the 16S rRNA gene by PCR-RFLP and PCR-DGGE. As a result, bacterial metagenomic DNAs with high quality, quantity and diversity were successfully extracted using a GeneJET kit and a chemical protocol. A sequence analysis of 73 representative DNA fragments from gels indicated a remarkable bacterial dysbiosis in all groups of diarrhea. Viral diarrhea was characterized by extremely reduced bacterial diversity with the blossom of Bifidobacterium and Streptococcus. Streptococcus was also the most abundant in persistent diarrhea. Beneficial bacteria that may play a role in the self- rebalance in intestinal bacterial communities, such as Bifidobacterium, Lactobacillus, and Enterococcus, were seen in all diarrheal groups, while Bacteroides and Akkermansia muciniphila were seen in the healthy group but absent in the diarrheal groups. This study provides additional evidence for a relationship between intestinal bacterial dysbiosis and diarrhea in children, emphasizing an increase in Streptococcus. [ABSTRACT FROM AUTHOR]

Simple Summary: This study reports the development of a triplex RT-qPCR assay for the simultaneous detection of Porcine Epidemic Diarrhea Virus (PEDV), Transmissible Gastroenteritis Virus (TGEV), and Porcine Kobuvirus (PKV) by targeting the conserved regions of their respective genes. The assay was optimized for specificity, sensitivity, and reproducibility, and demonstrated no cross-reactivity with other pathogens. It was validated against the Chinese standard single-assay method, exhibiting complete consistency in detecting these viruses in clinical samples. Finally, testing of 1502 clinical diarrhea samples revealed significant prevalence rates of PEDV, TGEV, and PKV, with mixed infections being common. The pathogens responsible for porcine viral diarrhea are diverse, causing significant economic losses to the pig industry. PEDV and TGEV are well-known pathogens causing diarrheal diseases in pigs, leading to significant economic losses in the breeding industry. In contrast, the newly identified diarrhea virus, PKV, has not garnered as much attention. However, co-infection of PKV with PEDV results in more severe symptoms in piglets, such as acute gastroenteritis, and promotes increased replication of PEDV. Rapid and accurate diagnosis of viral diarrhea is essential for farms to identify pathogens early and mitigate economic losses. This study describes the development of a triplex real-time fluorescent quantitative RT-qPCR technique that can simultaneously detect three RNA viruses associated with porcine viral diarrhea: PEDV, TGEV, and PKV. To establish the triplex RT-qPCR method for the simultaneous detection and identification of the above three diarrhea viruses, conserved regions of the M gene of TGEV, the N gene of PEDV, and the 3D gene of PKV were selected to design specific primers and probes. After optimizing the reaction conditions, the method's specificity, sensitivity, and reproducibility were evaluated. The triplex RT-qPCR method did not show a significant difference in PCR efficiency compared to the single RT-qPCR method. The method is specific to TGEV, PKV, and PEDV, exhibits no cross-reactivity with other pathogens, and demonstrates satisfactory sensitivity and reproducibility; the limit of detection (LOD) of PEDV, TGEV, and PKV is 11.42 copies/μL. Furthermore, the performance of the triplex RT-qPCR assay was compared with the Chinese standard single-assay method for detecting TGEV, PKV, and PEDV, showing complete consistency between the two methods (100% compliant). Subsequently, 1502 clinical diarrhea samples were collected from the Guangxi Zhuang Autonomous Region to investigate the local prevalence of TGEV, PKV, and PEDV and the positive rates were 16.38% (246/1502), 1.46% (22/1502), and 45.14% (678/1502), respectively. Co-infection of PEDV and PKV were most common, with a rate of 12.12% (182/1502). This study presents a valuable method for the rapid and simultaneous identification of PEDV, TGEV, and PKV in clinical animal farming practices, and provides a reassessment of the epidemiology of these diarrhea-causing viral pathogens in the Guangxi Zhuang Autonomous Region. [ABSTRACT FROM AUTHOR]

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]

This article discusses the mechanisms by which enteric viruses, such as rotavirus, norovirus, sapovirus, astrovirus, adenovirus, and poliovirus, induce diarrhea. The viruses disrupt intestinal homeostasis by infecting specific cells and causing structural changes, impairing nutrient absorption and leading to malabsorptive diarrhea. The enteric nervous system is also involved in diarrhea development, as viral infections activate serotonin secretion, leading to increased intestinal motility and fluid secretion. Viroporins, which are virally encoded proteins, play a role in manipulating cellular calcium signaling and facilitating viral replication, contributing to diarrhea. The article concludes by highlighting the importance of understanding these mechanisms for the development of antivirals and vaccines to reduce infection-associated diarrheal illness. [Extracted from the article]

Avian astrovirus can infect a variety of poultry species and cause viral diarrhea, with a wide epidemic range strong pathogenicity and a high incidence. Among them, Duck astrovirus 3(DAstV-3), Duck astrovirus 4(DAstV-4), Goose astrovirus 1(GoAstV-1) and Goose astrovirus 2(GoAstV-2) are four types of astroviruses newly discovered in waterfowl in recent years. In order to realize the rapid detection of these four kinds of waterfowl stellate viruses, specific primers and probes were engineered to target a highly conserved region of ORF1b gene of DAstV-3, GoAstV-1 and GoAstV-2 and the ORF2 gene of DAstV-4, and a quadruple fluorescence quantitative RT-PCR method was developed. The results indicate that the method established in this study has good specificity and no cross reactivity with other pathogens. This method can detect viruses with a minimum concentration of 1 × 101 copies/µL for DAstV-4, GoAstV-1 and GoAstV-2, respectively, while the minimum concentration for DAstV-3 is 1 × 102 copies/µL. Compared with the routinely used RT-PCR method, the limit of detection by the multiplex RT-PCR lower. Both intra- and inter-assay variability tests revealed excellent reproducibility. This method was then used to analyze 269 field samples, and the results were verified by genome sequencing. In conclusion, this study presents a sensitive, accurate, and specific method for detecting DAstV-3, DAstV-4, GoAstV-1, and GoAstV-2 in a single reaction, enabling the monitoring and differential diagnosis of these four types of waterfowl astroviruses. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and porcine rotavirus-A (PoRVA) are the four significant pathogens of viral diarrhea of piglets in large-scale pig farms, which have caused huge economic losses to the pig industry all over the world. Since these four viruses have very similar clinical symptoms, it is necessary to develop an excellent detection method that can differentiate and diagnose these four viruses. In this study, we developed a multiplex real-time RT-PCR method that can simultaneously differentiate and diagnose these four viruses. The method has high specificity and does not cross-react with other porcine viruses. The lower limit of detection was 2.18 × 102 copies/μL. In addition, we tested 97 clinical samples collected by this method, and the results were consistent with the detection results of traditional RT-PCR, indicating that this method is reliable. In summary, we developed a multiplex real-time RT-PCR method for simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA, and the results of this study can provide technical means for the differential diagnosis and epidemiological investigation of these four porcine diarrhea virus diseases. Porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and porcine rotavirus-A (PoRVA) are the four main pathogens that cause viral diarrhea in pigs, and they often occur in mixed infections, which are difficult to distinguish only according to clinical symptoms. Here, we developed a multiplex TaqMan-probe-based real-time RT-PCR method for the simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA for the first time. The specific primers and probes were designed for the M protein gene of PEDV, N protein gene of TGEV, N protein gene of PDCoV, and VP7 protein gene of PoRVA, and corresponding recombinant plasmids were constructed. The method showed extreme specificity, high sensitivity, and excellent repeatability; the limit of detection (LOD) can reach as low as 2.18 × 102 copies/μL in multiplex real-time RT-PCR assay. A total of 97 clinical samples were used to compare the results of the conventional reverse transcription PCR (RT-PCR) and this multiplex real-time RT-PCR for PEDV, TGEV, PDCoV, and PoRVA detection, and the results were 100% consistent. Subsequently, five randomly selected clinical samples that tested positive were sent for DNA sequencing verification, and the sequencing results showed consistency with the detection results of the conventional RT-PCR and our developed method in this study. In summary, this study developed a multiplex real-time RT-PCR method for simultaneous detection of PEDV, TGEV, PDCoV, and PoRVA, and the results of this study can provide technical means for the differential diagnosis and epidemiological investigation of these four porcine viral diarrheic diseases. [ABSTRACT FROM AUTHOR]

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The Porcine epidemic diarrhea virus (PEDV) presents a substantial risk to the domestic pig industry, resulting in extensive and fatal viral diarrhea among piglets. Recognizing the mucosal stimulation triggered by PEDV and harnessing the regulatory impact of lactobacilli on intestinal function, we have developed a lactobacillus-based vaccine that is carefully designed to elicit a strong mucosal immune response. Through bioinformatics analysis, we examined PEDV S proteins to identify B-cell linear epitopes that meet the criteria of being non-toxic, soluble, antigenic, and capable of neutralizing the virus. In this study, a genetically modified strain of Lactobacillus mucosae G01 (L.mucosae G01) was created by utilizing the S layer protein (SLP) as a scaffold for surface presentation. Chimeric immunodominant epitopes with neutralizing activity were incorporated at various sites on SLP. The successful expression of SLP chimeric immunodominant epitope 1 on the surface of L.mucosae G01 was confirmed through indirect immunofluorescence and transmission electron microscopy, revealing the formation of a transparent membrane. The findings demonstrate that the oral administration of L.mucosae G01, which expresses the SLP chimeric immunodominant gene epitope1, induces the production of secreted IgA in the intestine and feces of mice. Additionally, there is an elevation in IgG levels in the serum. Moreover, the levels of cytokines IL-2, IL-4, IFN-γ, and IL-17 are significantly increased compared to the negative control group. These results suggest that L. mucosae G01 has the ability to deliver exogenous antigens and elicit a specific mucosal immune response against PEDV. This investigation presents new possibilities for immunoprophylaxis against PEDV-induced diarrhea. [ABSTRACT FROM AUTHOR]

Porcine viral diarrhea is caused by many pathogens and can result in watery diarrhea, dehydration and death. Various detection methods, such as polymerase chain reaction (PCR) and real-time quantitative PCR (qPCR), have been widely used for molecular diagnosis. We developed a triplex real-time quantitative reverse transcription PCR (qRT-PCR) for the simultaneous detection of three RNA viruses potentially associated with porcine viral diarrhea: porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV), and porcine rotavirus A (PoRVA). The triplex qRT-PCR had R2 values of 0.999 for the standard curves of PEDV, TGEV and PoRVA. Importantly, the limits of detection for PEDV, TGEV and PoRVA were 10 copies/µL. The specificity test showed that the triplex qRT-PCR detected these three pathogens specifically, without crossreaction with other pathogens. In addition, the approach had good repeatability and reproducibility, with intra-and inter-assay coefficients of variation <1%. Finally, this approach was evaluated for its practicality in the field using 256 anal swab samples. The positive rates of PEDV, TGEV and PoRVA were 2.73% (7/256), 3.91% (10/256) and 19.14% (49/256), respectively. The co-infection rate of two or more pathogens was 2.73% (7/256). The new triplex qRT-PCR was compared with the triplex RT-PCR recommended by the Chinese national standard (GB/T 36871-2018) and showed 100% agreement for PEDV and TGEV and 95.70% for PoRVA. Therefore, the triplex qRT-PCR provided an accurate and sensitive method for identifying three potential RNA viruses for porcine viral diarrhea that could be applied to diagnosis, surveillance and epidemiological investigation. [ABSTRACT FROM AUTHOR]

Introduction: Porcine viral diarrhea is a common clinical disease, which results in highmortality and economic losses in the pig industry. Porcine epidemic diarrhea virus (PEDV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are important diarrhea viruses in pig herds. The similarities of their clinical symptoms and pathological changes make it difficult to distinguish these three viruses clinically. Therefore, there is a need for a highly sensitive and specific method to simultaneously detect and differentiate these viruses. Methods: A multiplex real-time PCR assay using TaqMan probes was developed to simultaneously detect PEDV, PoRV, and PDCoV. To assess the efficacy of the established assay, 30 clinical samples with diarrhea symptoms were used to compare the results obtained from the multiplex real-time PCR assay with those obtained from commercial singleplex real-time PCR kit. Importantly, a total of 4,800 diarrhea samples were tested and analyzed to validate the utility of the assay. Results: This multiplex real-time PCR assay showed high sensitivity, specificity, and excellent repeatability with a detection limit of 1 × 102 copies/µL. Comparing the results of the commercial singleplex real-time PCR kit and the multiplex real-time PCR method for detecting PEDV, PoRV, and PDCoV, there was complete agreement between the two approaches. Clinical data revealed single infection rates of 6.56% for PEDV, 21.69% for PoRV, and 6.65% for PDCoV. The co-infection rates were 11.83% for PEDV + PoRV, 0.29% for PEDV + PDCoV, 5.71% for PoRV + PDCoV, and 1.29% for PEDV + PDCoV + PoRV, respectively. Discussion: The multiplex real-time PCR method established in this study is a valuable diagnostic tool for simultaneously differentiating PEDV, PoRV, and PDCoV. This method is expected to significantly contribute to prevent and control the spread of infectious diseases, as well as aid in conducting epidemiological investigations. [ABSTRACT FROM AUTHOR]

Norovirus (NoV) genogroup II, polymerase type P31, capsid genotype 4, Sydney_2012 variant (GII.P31/GII.4_Sydney_2012) has been circulating at high levels for over a decade, raising the question of whether this strain is undergoing molecular alterations without demonstrating a substantial phylogenetic difference. Here, we applied next-generation sequencing to learn more about the genetic diversity of 14 GII.P31/GII.4_Sydney_2012 strains that caused epidemics in a specific region of Japan, with 12 from Kyoto and 2 from Shizuoka, between 2012 and 2022, with an emphasis on amino acid (aa) differences in all three ORFs. We found numerous notable aa alterations in antigenic locations in the capsid region (ORF2) as well as in other ORFs. In all three ORFs, earlier strains (2013–2016) remained phylogenetically distinct from later strains (2019–2022). This research is expected to shed light on the evolutionary properties of dominating GII.P31/GII.4_Sydney_2012 strains, which could provide useful information for viral diarrhea prevention and treatment. [ABSTRACT FROM AUTHOR]

Large-scale outbreaks of virus-associated severe diarrhea have occurred in pig populations since 2010. To investigate the prevalence and genetic evolution of the diarrhea-associated viruses responsible for the outbreaks, we tested 1,791 diarrhea samples collected from 213 pig farms in five provinces in southern China between 2021 and 2023. The test results showed that porcine epidemic diarrhea virus (PEDV) was the most frequently detected virus. The prevalence rates ranged from 47.40 to 52.22% in samples and 76.06% (162/213) in pig farms. Porcine rotavirus (PoRV) was the second common virus, with prevalence rates ranging from 25.81 to 50.81% in samples and 72.77%(155/213) in pig farms. Porcine delta coronavirus (PDCoV) was the third common virus, with prevalence rates ranging from 16.33 to 17.48% in samples and 38.50% (82/213) in pig farms. The detection rates of both transmissible gastroenteritis virus (TGEV) and porcine acute diarrheal syndrome coronavirus (SADS-CoV) were very low, less than 1.01% in samples and less than 3.76% in pig farms. In this study, we found SADS-CoV only in piglet diarrhea samples from Jiangxi, Guangdong, and Guangxi provinces in China, with a prevalence rate of 5.16% (11/213) in pig farms. Co-infection with these diarrhea-associated viruses is a common occurrence. The most common co-infections were PEDV and PoRV, with a prevalence rate of 6.64% (119/1,791), followed by PDCoV and PoRV, with a prevalence rate of 4.19% (75/1,791). Phylogenetic analyses showed that PEDV and PEDV variants prevalent in southern China during the past three years clustered into genotype GIIb and recombinant PEDV subtypes. Among the currently endemic PEDV, the most common mutations occurred in the collagenase equivalent (COE) and epitope regions of the spike gene. PoRV strains were mainly dominated by the G9 subtype, followed by the G5, G3 and G4 subtypes. Our results suggest that variant PEDV, PDCoV and PoRV are the main pathogens of swine diarrhea, and singular- or co-infection with pathogenic enteric CoV is common in pig herds in southern China. Therefore, prevention and control of porcine viral diarrhea should be given high attention. [ABSTRACT FROM AUTHOR]

Background: Diarrheal diseases caused by viral agents have led to a great morbidity, mortality, and economic loss in global pig industry. Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and group A porcine rotavirus (RVA) are main causative agents of swine viral diarrhea with similar clinical signs on Chinese farms and their co-infection is also common. However, it is still lack of a convenient method to detect these four agents. Methods: A TaqMan multiplex qPCR method was developed to detect PEDV, TGEV, PDCoV, and RVA, simultaneously. This method was then applied to investigate 7,342 swine fecal samples or rectal swabs, as well as 1,246 swine intestinal samples collected from 2075 farms in China in 2022. Results: Minimum detection limits of this method were 3 copies/µL for PEDV, 4 copies/µL for TGEV, 8 copies/µL for RVA, and 8 copies/µL for PDCoV, suggesting a good sensitivity. No signals were observed by using this method detecting other viral agents commonly prevalent in pigs, which is suggestive of a good specificity. Application of this method on investigating clinical samples demonstrated a relatively high positive rate for PEDV (22.21%, 1907/8588) and RVA (44.00%, 3779/8588). In addition, co-infection between PEDV and RVA was observed on 360 investigated farms, accounting for 17.35% (360/2075) of the farms where co-infection events were screened. Conclusions: A TaqMan multiplex qPCR method targeting PEDV, TGEV, PDCoV, and RVA was developed in this study. This method demonstrated a good specificity and sensitivity on investigating these four common viruses responsible for viral diarrhea on Chinese pig farms, which represents a convenient method for the monitoring and differential diagnosis of swine viral diarrhea. [ABSTRACT FROM AUTHOR]

BackgroundViral diarrhea is one of the major causes of morbidity and mortality in children. This study aimed to conduct etiological surveillance of viral diarrhea in Zhangzhou city, Fujian province, China, from 2017 to 2019 to identify the prevalence, distribution, and characteristics of viral pathogens causing gastrointestinal infections in the region.MethodsStool samples were collected from patients with acute diarrhea in Zhangzhou city, Fujian province, China, from 2017 to 2019. Rotavirus, norovirus, astrovirus, and adenovirus were detected using fluorescence immunochromatography assay.ResultsOf the total 5,627 samples that were collected, at least one of the viruses (rotavirus, norovirus, astrovirus and adenovirus) was found to be positive in 1,422 samples. Rotavirus, norovirus, astrovirus, and adenovirus, were detected in 53.73, 16.68, 15.52, and 14.97%, respectively. Mixed infections were determined in 17.65% of the positive samples. The predominant mixed infections observed were a combination of norovirus and astrovirus, followed by rotavirus and norovirus, and rotavirus and astrovirus. The highest positive rate was observed in the 12–23-month group for rotavirus and adenovirus, while a significantly higher positive rate was observed for norovirus and astrovirus in the 6–11-month group.ConclusionThese findings from this etiological surveillance highlight the significant burden of viral diarrhea in Zhangzhou city, with rotavirus being the predominant pathogen. The identification of common mixed infections provides insights into the complex nature of viral diarrhea transmission. Target interventions and public health strategies should be implemented, particularly during the winter and spring seasons, to prevent and control the spread of viral pathogens causing gastrointestinal infections in this region.

Human astrovirus (HAstV) is a known cause of viral gastroenteritis in children worldwide, but HAstV can cause also severe and systemic infections in immunocompromised patients. There are three clades of HAstV: classical, MLB, and VA/HMO. While all three clades are found in gastrointestinal samples, HAstV-VA/HMO is the main clade associated with meningitis and encephalitis in immunocompromised patients. To understand how the HAstV-VA/HMO can infect the central nervous system, we investigated its sequence-divergent capsid spike, which functions in cell attachment and may influence viral tropism. Here we report the high-resolution crystal structures of the HAstV-VA1 capsid spike from strains isolated from patients with gastrointestinal and neuronal disease. The HAstV-VA1 spike forms a dimer and shares a core beta-barrel structure with other astrovirus capsid spikes but is otherwise strikingly different, suggesting that HAstV-VA1 may utilize a different cell receptor, and an infection competition assay supports this hypothesis. Furthermore, by mapping the capsid protease cleavage site onto the structure, the maturation and assembly of the HAstV-VA1 capsid is revealed. Finally, comparison of gastrointestinal and neuronal HAstV-VA1 sequences, structures, and antigenicity suggests that neuronal HAstV-VA1 strains may have acquired immune escape mutations. Overall, our studies on the HAstV-VA1 capsid spike lay a foundation to further investigate the biology of HAstV-VA/HMO and to develop vaccines and therapeutics targeting it. Author summary: Human astroviruses, well-established as a leading cause of viral diarrhea, are increasingly associated with viral encephalitis in immunocompromised patients, with the sequence-divergent VA1 strains being the most common in these cases. Our study presents the high-resolution capsid spike structures from VA1 strains isolated from patients with both gastrointestinal and neuronal manifestations. Compared to the classical human astrovirus capsid spike structure, the VA1 capsid spike structure has striking differences in size, shape, and surface features, suggesting that the divergent VA1 strains may have evolved a different mechanism to attach to host cells, which in turn may influence their ability to infect the central nervous system. On the other hand, comparison of the gastrointestinal and neuronal VA1 spike structures reveals few differences. Instead, sequence and antigenic studies suggest that neuronal VA1 strains isolated from immunocompromised patients may have acquired mutations to escape immunoglobulin therapy. Altogether, this work provides a structural basis to investigate the mechanism of infection by the divergent human astrovirus VA1 strains and to develop tailored diagnostics, vaccines, and therapeutics against them. [ABSTRACT FROM AUTHOR]

Background: Porcine epidemic diarrhea (PED) is an infectious disease of the digestive tract caused by the porcine epidemic diarrhea virus (PEDV), characterized by vomiting, severe diarrhea, and high mortality rates in piglets. In recent years, the distribution of this disease in China has remarkably increased, and its pathogenicity has also increased. PEDV has been identified as the main cause of viral diarrhea in piglets. This study aimed to understand the genetic evolution and diversity of PEDV to provide a theoretical basis for the development of new vaccines and the prevention and treatment of PED. Methods: A PEDV strain was isolated from the small intestine of a diarrheal piglet using Vero cells. The virus was identified using reverse transcription-polymerase chain reaction (RT-PCR), indirect immunofluorescence assay (IFA), and transmission electron microscopy. The whole genome sequence was sequenced, phylogenetic analysis was conducted using MEGA (version 7.0), and recombination analysis was performed using RDP4 and SimPlot. The S protein amino acid sequence was aligned using Cluster X (version 2.0), and the S protein was modeled using SWISS-MODEL to compare differences in structure and antigenicity. Finally, the piglets were inoculated with PEDV to evaluate its pathogenicity in newborn piglets. Result: PEDV strain CH/HLJ/18 was isolated. CH/HLJ/18 shared 89.4–99.2% homology with 52 reference strains of PEDV belonging to the GII-a subgroup. It was a recombinant strain of PEDV BJ-2011-1 and PEDV CH_hubei_2016 with a breakpoint located in ORF1b. Unique amino acid deletions and mutations were observed in the CH/HLJ/18 S protein. The piglets then developed severe watery diarrhea and died within 7 d of inoculation with CH/HLJ/18, suggesting that CH/HLJ/18 was highly pathogenic to newborn piglets. Conclusion: A highly pathogenic recombinant PEDV GII-a strain, CH/HLJ/18, was identified in China, with unique deletion and mutation of amino acids in the S protein that may lead to changes in protein structure and antigenicity. These results will be crucial for understanding the prevalence and variation of PEDV and for preventing and controlling PED. [ABSTRACT FROM AUTHOR]

It is estimated that more than 4% of the causes of inpatient deaths in 2017 were due to diarrheal diseases. The current study is aimed to provide pooled prevalence of microorganisms causing diarrhea among Sudanese as well as determine any socio-cultural risk factors associated. A systematic review of the literature was conducted and regulated in accordance to PRISMA. After abstract and full text screening Twenty-one research articles were recruited to the study. Among witch eighteen research articles determined prevalence of causative microorganisms of Diarrhea; eight research articles determining prevalence of causative microorganisms of Diarrhea were conducted in Khartoum State, five in Gezira State, two in White Nile State, one in Kordofan State while two studies were conducted in several States. Moreover, majority of studies were concerned of prevalence among children while two studies were toward general population as well as mothers of children. The pooled prevalence of viral diarrhea in less than five years old children was 22.90% [15.37, 30.43] among more than 14 thousands' participants, the pooled prevalence of parasitic diarrhea was 31.40% [19.53, 43.27] among participants from different age groups while the pooled prevalence of bacterial diarrhea was 36.20% [14.00, 58.40]. No associated risk factors were able to be synthesized from included studies. [ABSTRACT FROM AUTHOR]

Human sapovirus (HuSaV) is a common cause of gastroenteritis worldwide and is responsible for approximately 4% of acute gastroenteritis episodes in Europe. As reported with norovirus, patients with immunocompromised states are at increased risk of developing HuSaV infection, which can lead to persistent diarrhea and chronic viral shedding in some individuals. Chronic infections are incompletely investigated in these patients, and, due to the lack of specific treatment for HuSaV infection, different clinical approaches were carried out in order to provide further evidence on clinical evolution of these patients with different treatments. In this retrospective study, we report five immunocompromised pediatric patients with recurrent diarrhea caused by HuSaV and long-term viral shedding. Stool samples were analyzed by real-time PCR and tested for enteropathogenic viruses and bacteria and protozoa. Among transplant recipients, reduction of immunosuppressant therapy led to clinical improvement and relief of symptoms, maintaining a balance between managing the infection and preventing graft rejection. Nitazoxanide for 14 days was only used in one of these patients, showing to be an effective therapy to achieve reduction in time to resolution of symptoms. Neither nitazoxanide nor modification of immunosuppressant therapy could avoid recurrences. Further investigations are needed to develop new approaches that can both clear the infection and avoid persistent diarrhea in these patients. [ABSTRACT FROM AUTHOR]

Lacticaseibacillus is one of the predominant microorganisms in gut from human and animal, and the lacticaseibacillus have effective applications against the viral diarrhea of piglets in the farm. However, the function and the concrete cell single pathways of the active ingredient from lacticaseibacillus was not clear within anti-infection in the postbiotics research. Here, we compared the biological function of extracellular polysaccharides (EPS) purified from lacticaseibacillus casei (L. casei) and gene editing lacticaseibacillus casei with the CRISPER-Cas9 technology, which were with the ability of antioxidation and anti-inflammation, and the EPS could also inhibit the ROS production within the Porcine Small Intestinal Epithelial Cells-J2 (IPEC-J2). Interestingly, we found that both of EPS and genome editing lacticaseibacillus casei could specifically target the IFN-λ expression in the IPEC-J2, which was beneficial against the PEDV infection in the virus replication and production with the qRT-PCR and indirect immunofluorescence methods. Finally, the STAT3 cell single pathway was stimulated to transcribe IFN-λ with the EPS to elucidate the detailed mechanism of activating type III IFN signals receptor of IL-10R2, which play the function between anti-inflammation and anti-virus in the PEDV infection. Taken together, our research linked a postbiotics of EPS with the antiviral infection of PEDV, which suggest that the lacticaseibacillus itself still have displayed the potential immunomodulatory activities, and highlight the immunomodulatory potential of EPS-producing microbes. [ABSTRACT FROM AUTHOR]

Transmissible gastroenteritis virus (TGEV) is an important swine enteric coronavirus causing viral diarrhea in pigs of all ages. Currently, the development of antiviral agents targeting host proteins to combat viral infection has received great attention. The heat shock protein 90 (HSP90) is a critical host factor and has important regulatory effects on the infection of various viruses. However, its roles in porcine coronavirus infection remain unclear. In this study, the effect of HSP90 on TGEV infection was evaluated. In addition, the influence of its inhibitor VER-82576 on proinflammatory cytokine (IL-6, IL-12, TNF-α, CXCL10, and CXCL11) production induced by TGEV infection was further analyzed. The results showed that the knockdown of HSP90AB1 and HSP90 inhibitor VER-82576 treatment resulted in a reduction in TGEV M gene mRNA levels, the N protein level, and virus titers in a dose-dependent manner, while the knockdown of HSP90AA1 and KW-2478 treatment had no significant effect on TGEV infection. A time-of-addition assay indicated that the inhibitory effect of VER-82576 on TGEV infection mainly occurred at the early stage of viral replication. Moreover, the TGEV-induced upregulation of proinflammatory cytokine (IL-6, IL-12, TNF-α, CXCL10, and CXCL11) expression was significantly inhibited by VER-82576. In summary, these findings indicated that HSP90AB1 is a host factor enhancing TGEV infection, and the HSP90 inhibitor VER-82576 could reduce TGEV infection and proinflammatory cytokine production, providing a new perspective for TGEV antiviral drug target design. [ABSTRACT FROM AUTHOR]

Enteric viruses like canine coronavirus (CCoV), canine parvovirus (CPV), and canine rotavirus (CRV) cause high morbidity and mortality in dogs, especially in young canines and hence pose great economic losses to dog breeders worldwide. This creates immense psychological distress and trauma for dog owners. CCoV is mistakenly diagnosed as CPV in many cases by clinicians though CCoV causes less severe disease than CPV. Sometimes both CCoV and CPV co-exist in a patient causing severe clinical disease and even death in dogs. In such cases, clinicians are uncertain about the prognosis. CRV is detected in several countries but no research has been carried out on it in Pakistan yet. The objective of this research was to observe if CCoV, CPV and CRV co-exist in diarrheic dogs. A total of 384 stool samples of dogs, that were reported at different clinics in Lahore, with the complaint of diarrhea, were collected and rendered for rapid detection of the viruses using immuno-chromatographic test kits followed by PCR-confirmation of the positive samples. The individual prevalence of CCoV, CPV and CRV turned out to be 21.6, 26.3 and 0% respectively. The prevalence of CCoV and CPV co-infection was 3.4%. Male German shepherd dogs under the age of 1 year are highly prone to get viral diarrhea in July and August. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine rotavirus (PoRV) is a major pathogen causing dehydrating diarrhea and fatality in newborn piglets, resulting in substantial economic losses in the swine industry worldwide. Therefore, the development of a safe and effective medication for treating PoRV infection is imperative. This study provides the first report on the anti-PoRV activity of Portulaca oleracea L. (POL). Our findings demonstrate that POL-P has antiviral activity against PoRV infection in vitro, and thus might be developed into a novel antiviral agent to control PoRV in pig farms. Diarrhea is one of the most common causes of death in young piglets. Porcine rotavirus (PoRV) belongs to the genus Rotavirus within the family Reoviridae, and is considered to be the primary pathogen causing diarrhea in piglets. Portulaca oleracea L. (POL) has been reported to alleviate diarrhea and viral infections. However, the antiviral effect of Portulaca oleracea L. polysaccharide (POL-P), an active component of POL, on PoRV infection remains unclear. This study demonstrated that the safe concentration range of POL-P in IPEC-J2 cells is 0–400 μg/mL. POL-P (400 μg/mL) effectively inhibits PoRV infection in IPEC-J2 cells, reducing the expression of rotavirus VP6 protein, mRNA and virus titer. Furthermore, on the basis of viral life cycle analysis, we showed that POL-P can decrease the expression of PoRV VP6 protein, mRNA, and virus titer during the internalization and replication stages of PoRV. POL-P exerts antiviral effects by increasing IFN-α expression and decreasing the expression levels of TNF-α, IL-6, and IL-10 inflammatory factors. Overall, our study found that POL-P is a promising candidate for anti-PoRV drugs. [ABSTRACT FROM AUTHOR]

Background and Aim: To meet domestic demand for meat, Indonesia imports live cattle from Australia, which have nonbovine viral diarrhea (BVD)-free status. The consequence of importing live cattle from Australia is potentially introducing a novel BVD variant to Indonesia. Therefore, detecting BVD early and determining the agent's characteristics and clinical symptoms are necessary. This study aimed to detect and characterize clinical symptoms of bovine viral diarrhea (BVD) and highlight the importance of farm management as a risk factor for the spread of BVD. This study aimed to provide information about the effectiveness of preventive measures against BVD in Australian-imported cattle at the Tanjung Priok Seaport Agricultural Quarantine. Bovine viral diarrhea is among the most common diarrheal diseases found in feedlots and is a severe health and economic problem in cattle. Materials and Methods: All cattle in a selected feedlot were examined for clinical symptoms on their first day of arrival. The sampling criteria included age, body weight, body temperature (BT), animal breath (AB), pulse (PL), conjunctivitis (CJ), hyperlacrimation (HL), hypersalivation (HS), DR, fever, limping leg (LL), emaciation, stomatitis (ST), weakness (WK), and coronitis (CR). In addition, 64 blood samples were taken from cattle that exhibited clinical symptoms of BVD. On the 3rd day of arrival, a blood sample showing positive clinical symptoms was examined using antigen (Ag)-capture enzymelinked immunosorbent assay (ELISA). The data from these clinical symptoms were analyzed alongside the laboratory results using multidimensional scale analysis, heatmap distribution, and principal component analysis (PCA). Furthermore, the positive serum samples obtained from the Ag-capture ELISA underwent a nested multiplex polymerase chain reaction and molecular detection and genetic characterization of BVDV based on the 5' untranslated region of the viral genome, followed by sequence and phylogenetic tree analyses. Results: Using PCA, 12 clinical symptom characteristics of BVD were determined from 13 clinical symptoms synergized with five cattle positive for Ag-capture ELISA. The clinical symptoms included internal factors such as physiological conditions of CJ, HL, HS, DR, BT, LL, loss of appetite, ST, WK, CR, AB, and PL. The screening test showed that five samples tested positive for the BVD Ag, while 59 tested negative. Phylogenetic tree analysis using a 360-nucleotide portion of the NS5B gene showed that Sample B23F5R had a distinct path compared to the other two samples in the phylogenetic diagram. The profile of sample B23F5R was closely related to BVDV reference subgenotype 1-a group (NCBI, access no. LC068605), with a homology percentage of 92.36%. Furthermore, this sample was similar to the BVDV reference 1-a, Strain 12, identified in Japan. The other two samples, B13F5R and A13F5R, showed close resemblance to the BVDV reference subgenotype 1-a that had been previously identified in Indonesia (NCBI, access no. MK411755), with homology percentages of 97.81% and 97.75%, respectively. Conclusion: The BVDV-1a strain is the main subtype present in beef cattle imported from Australia to West Java, Indonesia. The characteristics of clinical symptoms associated with BVD infection comprised 12 symptoms synergized with the positive sample in the PCA. The present results can facilitate the development of preventive and control measures for BVD circulation in Indonesia. [ABSTRACT FROM AUTHOR]

Background Antibiotics are commonly overused for diarrheal illness in many low- and middle-income countries, partly due to a lack of diagnostics to identify viral cases, in which antibiotics are not beneficial. This study aimed to develop clinical prediction models to predict risk of viral-only diarrhea across all ages, using routinely collected demographic and clinical variables. Methods We used a derivation dataset from 10 hospitals across Bangladesh and a separate validation dataset from the icddr,b Dhaka Hospital. The primary outcome was viral-only etiology determined by stool quantitative polymerase chain reaction. Multivariable logistic regression models were fit and externally validated; discrimination was quantified using area under the receiver operating characteristic curve (AUC) and calibration assessed using calibration plots. Results Viral-only diarrhea was common in all age groups (<1 year, 41.4%; 18–55 years, 17.7%). A forward stepwise model had AUC of 0.82 (95% confidence interval [CI],.80–.84) while a simplified model with age, abdominal pain, and bloody stool had AUC of 0.81 (95% CI,.78–.82). In external validation, the models performed adequately although less robustly (AUC, 0.72 [95% CI,.70–.74]). Conclusions Prediction models consisting of 3 routinely collected variables can accurately predict viral-only diarrhea in patients of all ages in Bangladesh and may help support efforts to reduce inappropriate antibiotic use. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine epidemic diarrhea virus (PEDV), porcine bocavirus (PBoV), and porcine rotavirus (PoRV) are associated with porcine viral diarrhea. The three viruses are often comorbid in mixed infection; therefore, it is necessary to develop a specific and rapid method for simultaneous detection. In this study, a multiple detection method, loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick (LFD), was established to detect three pathogens in 125 field samples. The consistency between the real-time fluorescence quantitative PCR (rt-qPCR) and the triplex LAMP–LFD assay was over 99%. The assay has been proven to be highly specific and sensitive. Porcine epidemic diarrhea virus (PEDV), porcine bocavirus (PBoV), and porcine rotavirus (PoRV) are associated with porcine viral diarrhea. In this study, triplex loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick (LFD) was established for the simultaneous detection of PEDV, PoRV, and PBoV. The PEDV-gp6, PoRV-vp6, and PBoV-vp1 genes were selected to design LAMP primers. The amplification could be carried out at 64 °C using a miniature metal bath within 30 min. The triplex LAMP–LFD assay exhibited no cross-reactions with other porcine pathogens. The limits of detection (LODs) of PEDV, PoRV, and PBoV were 2.40 × 101 copies/μL, 2.89 × 101 copies/μL, and 2.52 × 101 copies/μL, respectively. The consistency between rt-qPCR and the triplex LAMP–LFD was over 99% in field samples testing. In general, the triplex LAMP–LFD assay was suitable for the rapid and simultaneous detection of the three viruses in the field. [ABSTRACT FROM AUTHOR]

Porcine viral diarrhea is very common in clinical practice and has caused huge losses to the pig industry. Porcine epidemic diarrhea virus (PEDV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are important pathogens of porcine viral diarrhea. Co-infection situations among these three viruses in clinics are common, which increases the difficulty of differential diagnosis. Currently, polymerase chain reaction (PCR) is commonly used to detect pathogens. TaqMan real-time PCR is more sensitive than conventional PCR and has better specificity and accuracy. In this study, a triplex real-time RT-PCR assay based on TaqMan probes was developed for differential detection of PEDV, PoRV, and PDCoV. The triplex real-time RT-PCR assay developed in this study could not detect unrelated pathogens and showed satisfactory specificity, sensitivity, repeatability, and reproducibility with a limit of detection (LOD) of 6.0 × 101 copies/μL. Sixteen clinical samples were used to compare the results of the commercial RT-PCR kit and the triplex RT-PCR for PEDV, PoRV, and PDCoV detection, and the results were completely consistent. A total of 112 piglet diarrhea samples collected from Jiangsu province were next used to study the local prevalence of PEDV, PoRV, and PDCoV. The positive rates of PEDV, PoRV, and PDCoV detected by the triplex real-time RT-PCR were 51.79% (58/112), 59.82% (67/112), and 2.68% (3/112), respectively. The co-infections of PEDV and PoRV were frequent (26/112, 23.21%), followed by the co-infections of PDCoV and PoRV (2/112, 1.79%). This study established a useful tool for simultaneous differentiation of PEDV, PoRV, and PDCoV in practice and provided valuable information on the prevalence of these diarrhea viral pathogens in Jiangsu province. [ABSTRACT FROM AUTHOR]

Bovine viral diarrhea (BVD) is one of the most imperative worldwide diseases in ruminants leading to substantial damage as well as extensive economic losses for the cattle industry. To our knowledge, there have been no reports about the disease's situation in New Valley province, so the goal of this study was to estimate the level of bovine viral diarrhea virus (BVDV) infection in cattle herds through serodiagnosis and determining factors influencing the course of infection (current epidemiological situation) which will aid in establishment of control strategy. In this study a total of 1400 cattle of different ages, sexes and breeds (not previously vaccinated with BVD vaccines) were screened clinically for symptoms of BVD infection. 260 cattle out of totally 1400 screened cattle (18.6%) showing clinical signs arousing suspicion of BVD (49 cattle (18.8%) exhibit respiratory manifestation, 63 (24.2%) cattle suffered from diarrhea, 37 (14.2%) with mixed enteric and respiratory signs, 43 (16.5%) with retarded growth, oral lesion was detected in only 6 cattle (2.3%), While the congenital anomalies were recorded in 18 (6.9%), abortion occurred in 29 female cattle (11.1%) and only 15 cattle (5.8%) suffered from infertility trouble). Totally 260 clinically diseased cattle undergo Serum Neutralization Test (SNT) for detection of BVDV infection. The results revealed that 83 cattle out of 260 (31.9%) were detected as seropositive (positive for BVDV antibodies), showed clinical signs of BVD infection represented in 17 cattle showed respiratory manifestation (nasal discharge), 22 cattle suffered from diarrhea, 12 with mixed enteric and respiratory signs, 6 with retarded growth, oral lesion was detected in Only 2 cattle, While the congenital anomalies were recorded in 5, abortion occurred in 15 female cattle and only 4 cattle suffered from Infertility trouble According to age, the seroprevalence of BVDV for antibodies was higher in in cattle aged (>6M to 2 year) (37.50%) and the lowest rate was (21.73%) in cattle aged (1day: 2month). Rendering to sex and breed, female cattle (37.80%) had a greater infection rate than male (21.87%) and infection was higher in Frisian bread (34.05%) than native bread (29.50%). Concerning to season, the highest prevalence of infection was (40.24%) in Winter and the lowest rate was (16.66%) in Autumn. Regarding to locality, the highest prevalence of infection was (45.45%) in EL-Farfara, and the lowest rate was (31%) in Balat. This prevalence of BVDV in cattle of new valley province, Egypt reveals the necessity of further studying BVDV infection in the area. [ABSTRACT FROM AUTHOR]

An important factor affecting the performance of cattle is the satisfaction of animal needs in nutrients, macro- and microelements. High-yielding dairy cows with intense metabolism are especially sensitive to errors in feeding practices, because even minor nutritional or mineral deficiencies cause metabolic disorders leading to immunodeficiency conditions, reduced resistance to infectious agents and raised sensitivity to infections with pathogenic and opportunistic microorganisms. Besides some infections, for example acute respiratory diseases, also result in metabolic disorders, immunosuppression, decrease in the overall resistance of the organism and ultimately in mixed infections. All this is the reason for low efficacy of vaccination against major infectious diseases. The paper presents the results of testing of plant tissue product «Vidoral», developed by the FSBEI HE Ural SAU for prevention of immunodeficiency conditions in cattle, caused by different factors. The product was injected subcutaneously at a dose of 0.025 ml/kg of live weight to cows in late gestation and calves showing mild anemia, low red blood cells counts, leukocytosis, high lymphocyte and monocyte counts and elevated erythrocyte sedimentation rate, which suggest inflammatory processes in the animal body. Fourteen days post injection blood was collected for hematological and biochemical testing. It was demonstrated that the product restores the animal metabolism, improves hematopoiesis and reduces inflammation. It was established that injection of «Vidoral» in combination with vaccination against infectious rhinotracheitis, viral diarrhea, infection with respiratory syncytial virus and parainfluenza-3 virus induced generation of specific antibodies against the abovementioned infections.

ObjectiveTo examine infection characteristics and genetic evolution of adenovirus isolated in patients with diarrhea in Ningxia Hui Autonomous Region (Ningxia) for providing evidence to the prevention and control of human adenovirus infection. MethodsTotally 6 695 fecal specimens were collected from diarrheal outpatients at 10 diarrhea sentinel hospitals in 5 cities of Ningxia from 2015 through 2020. Adenovirus positive samples were screened out with real-time PCR. The gene of adenovirus hexon was amplified with PCR and sequenced for virus type identification and phylogenetic analysis. ResultsOf all the specimens detected, 4.2% (278) were adenovirus positive and the positive rate was the highest (7.7%) for the specimens collected in 2017. There were significant differences in the adenovirus positive rate among the specimens collected in different seasons and isolated from the outpatients of various age groups. Gene sequences were obtained for the 80 adenovirus strains isolated from the 278 viral positive specimens. For the 80 viral strains, three genotypes were identified and the main type was F41, followed by type F40 and type C1.ConclusionAdenovirus-induced diarrhea occured across a year among the people of various ages in Ningxia during 2015 – 2020 and the dominant adenovirus strain causing the diarrheal diseases was type F41.

Introduction: Iron deficiency is the most common micronutrient deficiency worldwide. Iron activates the growth and differentiation of immune cells and cytokine actions. Interleukin 6 is a pro-inflammatory cytokine that stimulates the differentiation of B lymphocytes into plasma cells and activates T lymphocytes. Our study aimed to evaluate the correlation between blood levels of iron and interleukin 6 (IL-6) and its impact on infection rates in children. Materials and methods: The study included 36 children. The serum concentration of IL-6, iron, and morphology parameters from a venous blood sample were assessed. An anonymous survey of children’s parents was conducted regarding the frequency of infections before and after the diagnosis of anemia. Results: In the study group, the levels of IL-6 were higher than in the control group and showed greater variability between individual patients. There was a statistically significant, negative correlation between IL-6 and iron levels within the study group (p = 0.012), and a decrease in the number of viral diarrheas after the diagnosis of anemia compared to the state before diagnosis (p = 0.041). There was no statistically significant change in the number of colds or the mean duration of infection (p = 0.144, p = 0.498), nor in the incidence of other infectious diseases and antibiotics intake in the study group before and after iron deficiency anemia diagnosis (p = 0.500, p = 0.219). Conclusions: IL-6 might play a role in iron deficiency anemia. Increased levels of IL-6 were shown not to correspond with visible changes in rates and the course of colds, but did result in a reduction in the frequency of viral diarrhea. Further research on a larger group of carefully selected patients is required to determine the effect of anemia on the immune system. [ABSTRACT FROM AUTHOR]

Objective To investigate the effects of bifidobacterium triple viable bacteria combined with recombinant human interferon α-1b injection on neutrophil rod nucleus, lobular nucleus and serum myocardial zymogram in children patients with viral diarrhea.Methods Ninety-three children patients with viral diarrhea in this hospital from April 2017 to December 2019 were retrospectively collected as the study subjects and divided into two groups according to the treatment regimen.The two groups were given the routine treatment. On this basis, 46 cases treated with bifidobacterium triple viable bacteria served as the control group, and 47 cases treated by bifidobacterium triple viable bacteria combined with recombinant human interferon α-1b injection as the observation group.The clinical efficacy, symptom relief time, neutrophil rod nucleus, lobular nucleus, serum inflammatory cytokines (IL-6, IL-8, TNF-α), myocardial zymogram [creatine kinase (CK), creatine kinase isoenzyme (CK-MB), hydroxybutyrate dehydrogenase (HBDH), lactate dehydrogenase (LDH)] and immune function before and after treatment were compared between the two groups.Results The total effective rate in the observation group was 95.74% (45/47), which was higher than 80.43% (37/46) in the control group, and the difference was statistically significant (P <0.05).The antiemetic, antidiarrheal and antipyretic time of the observation group was shorter than that of the control group (P <0.05).The count of neutrophil rod nucleus and lobulated nucleus after 1, 3 d treatment in the observation group was lower than that in the control group, and the differences were statistically significant (P <0.05).The serum inflammatory factors IL-6, IL-8 and TNF-α levels after 1, 3 d treatment in the observation group was lower than those in the control group (P <0.05).Serum CK, CK-MB, LDH and HBDH levels after 1, 3 d treatment in the observation group were lower than those in the control group, and the differences were statistically significant (P <0.05).The CD4+ and CD4+/CD8+ levels after 1, 3 d treatment in the observation group were higher than those in the control group, the CD8+ level was lower than that in the control group, and the differences were statistically significant(P <0.05).Conclusion Bifidobacterium triple viable bacteria combined with recombinant human interferon α-1b is conducive to enhance the treatment effect and shorten the symptom improvement time in the children patients with viral diarrhea by reducing the level of myocardial zymogram and regulating immune inflammatory response. [ABSTRACT FROM AUTHOR]

Porcine epidemic diarrhea virus (PEDV) remains one of the major causative microorganisms of viral diarrhea in piglets worldwide, with no approved drugs for treatment. We identified a natural molecule, flavonol, which is widely found in tea, vegetables and herbs. Subsequently, the antiviral activity of compound flavonol was evaluated in Vero cells and IPEC-J2 cells, and its anti-PEDV mechanism was analyzed by molecular docking and molecular dynamics. The results showed that flavonol could effectively inhibit viral progeny production, RNA synthesis and protein expression of PEDV strains in a dose-dependent manner. When flavonol was added simultaneously with viral infection in Vero cells, it demonstrated potent anti-PEDV activity by affecting the viral attachment and internalization phases. Similarly, in IPEC-J2 cells, flavonol effectively inhibited PEDV infection at different stages of infection, except for the release phase. Moreover, flavonol mainly interacts with PEDV Mpro through hydrogen bonds and hydrophobic forces, and the complex formed by it has high stability. Importantly, flavonol also showed broad-spectrum activity against other porcine enteric coronaviruses such as TGEV and PDCoV in vitro. These findings suggest that flavonol may exert antiviral effects by interacting with viral Mpro, thereby affecting viral replication. This means that flavonol is expected to become a potential drug to prevent or treat porcine enteric coronavirus. • PEDV induces distinct morphological changes in both Vero cells and IPEC-J2 cells. • Flavonol primarily disrupts the initial stage of PEDV infection in Vero cells. • Flavonol prevents the advancement of multiple life cycle phases of PEDV propagation in IPEC-J2 cells. • Flavonol exhibits a wide-ranging efficacy against porcine enteric coronavirus activity. [ABSTRACT FROM AUTHOR]

Background: Human norovirus (HuNoV) is the leading viral cause of diarrhea, with GII.4 as the predominant genotype of HuNoV outbreaks globally. However, new genogroup variants emerge periodically, complicating the development of anti-HuNoV vaccines; other prophylactic or therapeutic medications specifically for HuNoV disease are lacking. Passive immunization using oral anti-HuNoV antibodies may be a rational alternative. Here, we explore the feasibility of using avian immunoglobulins (IgY) for preventing HuNoV infection in vitro in a human intestinal enteroid (HIE) model. Methods: Hens were immunized with virus-like particles (VLP) of a GII.4 HuNoV strain (GII.4/CHDC2094/1974/US) by intramuscular injection. The resulting IgY was evaluated for inhibition of binding to histo-blood group antigens (HBGA) and viral neutralization against representative GII.4 and GII.6 clinical isolates, using an HIE model. Results: IgY titers were detected by three weeks following initial immunization, persisting at levels of 1:221 (1:2,097,152) from 9 weeks to 23 weeks. Anti-HuNoV IgY significantly (p < 0.05) blocked VLP adhesion to HBGA up to 1:12,048 dilution (0.005 mg/mL), and significantly (p < 0.05) inhibited replication of HuNoV GII.4[P16] Sydney 2012 in HIEs up to 1:128 dilution (0.08 mg/mL). Neutralization was not detected against genotype GII.6. Conclusions: We demonstrate the feasibility of IgY for preventing infection of HIE by HuNoV GII.4. Clinical preparations should cover multiple circulating HuNoV genotypes for comprehensive effects. Plans for animal studies are underway. [ABSTRACT FROM AUTHOR]

Rotavirus A (RVA) is an important pathogen causing acute gastroenteritis in animals and humans. Attachment to the host receptor is a crucial step for virus replication. The VP8* domain is the distal terminal region of the RVA spike protein VP4 (expressed by the P gene) and is important for rotavirus binding and infectivity. Recent studies have indicated a role for non-sialylated glycans, including mucin core 2 and histo-blood group antigens (HBGAs), in the infectivity of human and animal group A rotaviruses. In the present study, we determined if porcine rotavirus-derived recombinant VP8* of the endemic strains 14R103 G5P[6], 13R054 G5P[7], 12R010 G4P[13], 12R046 G9P[23], and 12R022 G2P[27] interact with hitherto uncharacterized glycans. We successfully produced five recombinant GST-VP8* proteins of genotype P[6], P[7], P[13], P[23], and P[27]. The hemagglutination assay showed genotypes P[7] and P[23] hemagglutinate porcine and human red blood cells. In an array screen of > 300 glycans, recombinant VP8* of rotavirus genotype P[6], P[7], and P[13] showed specific binding to glycans with a Gal-β-1,4-Glc (β-lactose) motif, which forms the core structure of HBGAs. The specificity of glycan-binding was confirmed through an ELISA-based oligosaccharide binding assay. Further, 13R054 G5P[7] and 12R046 G9P[23] infectivity was significantly reduced by β-lactose in MA104 cells and primary porcine enterocytes. These data suggest that lactose, the main natural sugar in milk, plays an important role in protecting piglets from enteric viral replication and diarrhea. [ABSTRACT FROM AUTHOR]

Porcine viral diarrhea diseases affect the swine industry, resulting in significant economic losses. Porcine epidemic diarrhea virus (PEDV) genotypes G1 and G2, and groups A and C of the porcine rotavirus, are major etiological agents of severe gastroenteritis and profuse diarrhea, particularly among piglets, with mortality rates of up to 100%. Based on the high prevalence rate and frequent co-infection of PEDV, RVA, and RVC, close monitoring is necessary to avoid greater economic losses. We have developed a multiplex TaqMan probe-based real-time PCR for the rapid simultaneous detection and differentiation of PEDV subtypes G1 and G2, RVA, and RVC. This test is highly sensitive, as the detection limits were 20 and 100 copies/μL for the G1 and G2 subtypes of PEDV, respectively, and 50 copies/μL for RVA and RVC, respectively. Eighty-eight swine clinical samples were used to evaluate this new test. The results were 100% in concordance with the standard methods. Since reassortment between porcine and human rotaviruses has been reported, this multiplex test not only provides a basis for the management of swine diarrheal viruses, but also has the potential to impact public health as well. [ABSTRACT FROM AUTHOR]

Viral diarrhea is one of the leading causes of morbidity and mortality in children. This study was conducted to disclose the etiological cause and epidemiological features of viral diarrhea among children in China. From 2009 to 2021, active surveillance was performed on pediatric patients with acute diarrhea and tested for five enteric viruses. Positive detection was determined in 65.56% (3325/5072) patients and an age‐specific infection pattern was observed. A significantly higher positive rate was observed in 12–23‐month‐old children for rotavirus (47.46%) and adenovirus (7.06%), while a significantly higher positive rate was observed for norovirus (37.62%) in 6–11‐month‐old patients, and for astrovirus (11.60%) and sapovirus (10.79%) in 24–47‐month‐old patients. A higher positive rate of rotavirus in girls and norovirus in boys was observed only among 6–11 months of patients. We also observed more norovirus among patients from rural areas in the 0–5‐ and 36–47‐month groups and more rotavirus among those from rural areas in the 12–23‐month group. Diarrhea severity was greater for rotavirus in the 6–23‐month group and norovirus in the 6–11‐month group. Coinfections were observed in 29.26% (973/3325) of positive patients, and were most frequently observed between rotavirus and others (89.31%). Our findings could help the prediction, prevention, and potential therapeutic approaches to viral diarrhea in children. [ABSTRACT FROM AUTHOR]

In Western countries during the post-World War II decades, endemic viral diseases were increasingly important to health. Such diseases have attracted limited historical attention. Due to changing methods of livestock production, they were particularly prevalent on the farm. This article uses a case study of the cattle disease, bovine viral diarrhoea (BVD), to demonstrate their historical significance. Spanning North America, the UK and Australia, it reveals the complex nature of BVD, and how and why its clinical, aetiological, epidemiological and host species identities evolved over time. This analysis sheds new light on how endemic viral diseases of livestock were experienced, understood and distributed in this period, and the influence exerted by changing agricultural practices, concerns about biological warfare and the development of virology as applied to veterinary medicine. [ABSTRACT FROM AUTHOR]

Porcine bocavirus (PBoV) was first identified in Sweden in 2009. Due to its association with healthy as well as diseased pigs, its role in clinical disease has not been reported yet. In the present study, bocavirus was identified from the intestinal content of a 30-day-old piglet and its whole genome was constructed and phylogenetic analysis was carried on. The pathogenesis of bocavirus was investigated following orogastric inoculation of the colostrum-deprived newborn piglet with bacteria free intestinal content. The bocavirus-inoculated piglets developed diarrhea, shed virus in the rectal swabs from 18 h post inoculation and developed macroscopic and microscopic lesions in small intestine with virus confirmed by conventional PCR. This study experimentally confirmed pathogenicity and characterized bocavirus as the etiological agent of diarrhea in the colostrum-deprived newborn piglets. On phylogenetic analysis, it was observed that this virus has long evolutionary history with subsequent mutation as well as better host adaptation. This study highlights the importance of identifying bocavirus as the etiological agent of viral diarrhea that could threaten livestock, public health as well as economic loss. • A porcine bocavirus (PBoV/HB/30/2018) associated with diarrhea was discovered in China. • Pathogenicity of PBoV/HB/30/2018 was evaluated in colostrum-deprived newborn piglets. • Complete genome of PBoV/HB/30/2018 was sequenced and assembled. • The origin and phylogeny of PBoV/HB/30/2018 were investigated. [ABSTRACT FROM AUTHOR]

Diarrhea is one of the leading causes of death among children, especially in the age under 5, but few studies are available on viral diarrhea in Shenyang. To understand the infection status and the relevant epidemiological characteristics of viral diarrhea and to fill gaps of the distribution of viruses across Shenyang in children under the age of 5 with diarrhea, from 2018 to 2020, stool specimens of children with diarrhea aged 0–59 months and surveillance data were collected in Sentinel Hospital of Shenyang. Specimens were then tested to determine the type of viruses, the seasonal and spatial patterns for major viruses were determined. Viruses were identified in 47.9% of the 897 samples from children with diarrhea. The main viruses of stool samples were rotavirus (16.9%, predominant type G9P[8]), calicivirus (14.7%), adenovirus (11.8%), and astrovirus (4.5%). Viral infections were mainly detected in the age of 0–12 months. In the area of Shenyang, Huanggu had the most cases (198, 22.1%), followed by Dadong (137, 15.3%) and Hunnan (135, 15.1%). The positive rate of viruses varied among patients of different ages, seasons, and regions. Public health entities and the government should develop corresponding measures for different age groups, seasons, and regions. Highlights: Occurrence of enteric virus infections in cases of virus diarrhea.Rotavirus is the predominant viral agents infecting hospitalized children under the age of five.Infection rate of viruses varied over the place in both urban and rural areas of Shenyang. [ABSTRACT FROM AUTHOR]

Comprehensive data on bacterial and viral pathogens of diarrhea and studies applying culture-independent methods for examining antibiotic resistance in wastewater are lacking. This study aimed to simultaneously quantify antibiotic resistance genes (ARGs), class 1 integron-integrase (int1), bacterial and viral pathogens of diarrhea, 16S rRNA, and other indicators using a high-throughput quantitative PCR (HT-qPCR) system. Thirty-six grab wastewater samples from a wastewater treatment plant in Japan, collected three times a month between August 2022 and July 2023, were centrifuged, followed by nucleic acid extraction, reverse transcription, and HT-qPCR. Fourteen targets were included, and HT-qPCR was performed on the Biomark X9™ System (Standard BioTools). For all qPCR assays, R2 was ≥0.978 and the efficiencies ranged from 90.5% to 117.7%, exhibiting high performance. Of the 36 samples, 20 (56%) were positive for Norovirus genogroup II (NoV-GII), whereas Salmonella spp. and Campylobacter jejuni were detected in 24 (67%) and Campylobacter coli in 13 (36%) samples, with mean concentrations ranging from 3.2 ± 0.8 to 4.7 ± 0.3 log 10 copies/L. NoV-GII detection ratios and concentrations were higher in winter and spring. None of the pathogens of diarrhea correlated with acute gastroenteritis cases, except for NoV-GII, suggesting the need for data on specific bacterial infections to validate bacterial wastewater-based epidemiology (WBE). All samples tested positive for sul1 , int1 , and bla CTX-M , irrespective of season. The less explored bla NDM-1 showed a wide prevalence (>83%) and consistent abundance ranging from 4.3 ± 1.0 to 4.9 ± 0.2 log 10 copies/L in all seasons. sul1 was the predominant ARG, whereas absolute abundances of 16S rRNA, int1 , and bla CTX-M varied seasonally. int1 was significantly correlated with bla CTX-M in autumn and spring, whereas it showed no correlation with bla NDM-1 , questioning the applicability of int1 as a sole indicator of overall resistance determinants. This study exhibited that the HT-qPCR system is pivotal for WBE. [Display omitted] • HT-qPCR system successfully analyzed pathogens and ARGs in a single run. • NoV-GII concentration was linked with AGE cases but not bacterial pathogens. • Specific bacterial infection data could be useful for validation of bacterial WBE. • sul1 , bla CTX-M , an ESBL, and the less explored bla NDM-1 were widely prevalent. • int1 correlated with bla CTX-M in some seasons and did not correlate with bla NDM-1. [ABSTRACT FROM AUTHOR]

Canine parvovirus (CPV) is the main cause of viral diarrhea in dogs. CPV became a global disease in 1978 and was endemic all over the world. CPV-2 was the first strain to be identified, but with genetic mutations, new genotypes such as CPV-2a/2b/2c/new-2a/new-2b have emerged. In this study, 128 fecal samples of stray dogs suspected of CPV-2 infection were collected from January to March 2021 in Shanghai, China. All samples were screened by PCR and further analyzed by VP2 gene. The positive rate of CPV-2 was 9.4% (12/128), of which 6 CPV-2 isolates were successfully isolated. Phylogenetic tree analysis showed that 4 isolates were CPV-2c genotype and 2 were new-CPV-2b genotype. VP-2 is a key protein that determines the antigenic properties, host range and receptor binding of cpv-2. The results of VP2 amino acid sequence analysis in this study showed that the CPV-2c isolated strain was the same as the previous strains reported in China, including F267Y, Y324I, Q370R and A5G mutations in addition to the typical N426E mutations. Similarly, in addition to the conventional N426D, S297A, F267Y and Y324I mutations, the new CPV-2b isolate also had a new mutation of T440A. This study further confirmed the prevalence of CPV-2c and new-CPV-2b in Shanghai, and also found a new mutation site of new-CPV-2c, which provided a theoretical basis for further enriching the epidemiological data of CPV-2 in Shanghai, as well as the development of vaccines and the prevention and control of the disease. • High prevalence of CPV-2 in stray dogs. • CPV-2c and new-CPV-2b co-exist and circulate in Stray dogs locally in Shanghai, China. • Current Vaccines may not be able to prevent the CPV prevalence by the mutated strains. [ABSTRACT FROM AUTHOR]

In this report, a multiplex PCR method was developed for the detection of three diarrhea-associated viruses in mink, including circovirus (MCV), bocavirus (MBoV), and enteritis virus (MEV). Three compatible sets of primers specific for each virus were designed respectively based on their conserved sequences. After optimization of the crucial factors such as primer concentration and annealing temperature in single and multiple amplification, three specific fragments were simultaneously amplified with the highest sensitivity and specificity in one PCR reaction. The fragments amplified were 259 bp (MCV)，455 bp (MBoV) and 671 bp (MEV). The sensibility of this one-step multiplex PCR is about 10 times lower than that of regular singleplex PCR. There were no cross-reactions with some relevant pathogens like mink coronavirus, canine distemper virus, and aleutian mink disease virus. In our study we analyzed viral DNA in mink fecal samples by multiplex PCR assay from China, which revealed the occurrence of MCV, MBoV, and MEV as 3.1 %, 5.7 %, and 9.8 %, respectively. The testing results of multiplex PCR agreed with the singleplex PCR results with a coincidence rate of 100 %. These results indicated that the method could provide technical support for rapid detection of the three diarrhea-associated viruses, and epidemiological investigation of mink viral diarrhea. ● A multiplex PCR assay was developed to simultaneously detect the DNA of MCV, MBoV, and MEV. ● The sensitivity of multiplex PCR for MCV, MBoV, and MEV was 1,020 copies, 1,020 copies, and 858 copies, respectively. ● The assay was specific for detecting one or more of the viruses. [ABSTRACT FROM AUTHOR]

Porcine viral diarrhea is a common ailment in clinical settings, causing significant economic losses to the swine industry. Notable culprits behind porcine viral diarrhea encompass transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), porcine deltacoronavirus (PDCoV), and porcine rotavirus-A (PoRVA). Co-infections involving the viruses are a common occurrence in clinical settings, thereby amplifying the complexities associated with differential diagnosis. As a consequence, it is therefore necessary to develop a method that can detect and differentiate all four porcine diarrhea viruses (TGEV, PEDV, PDCoV, and PoRVA) with a high sensitivity and specificity. Presently, polymerase chain reaction (PCR) is the go-to method for pathogen detection. In comparison to conventional PCR, TaqMan real-time PCR offers heightened sensitivity, superior specificity, and enhanced accuracy. This study aimed to develop a quadruplex real-time RT-qPCR assay, utilizing TaqMan probes, for the distinctive detection of TGEV, PEDV, PDCoV, and PoRVA. The quadruplex real-time RT-qPCR assay, as devised in this study, exhibited the capacity to avoid the detection of unrelated pathogens and demonstrated commendable specificity, sensitivity, repeatability, and reproducibility, boasting a limit of detection (LOD) of 27 copies/μL. In a comparative analysis involving 5483 clinical samples, the results from the commercial RT-qPCR kit and the quadruplex RT-qPCR for TGEV, PEDV, PDCoV, and PoRVA detection were entirely consistent. Following sample collection from October to March in Guangxi Zhuang Autonomous Region, we assessed the prevalence of TGEV, PEDV, PDCoV, and PoRVA in piglet diarrhea samples, revealing positive detection rates of 0.2 % (11/5483), 8.82 % (485/5483), 1.22 % (67/5483), and 4.94 % (271/5483), respectively. The co-infection rates of PEDV/PoRVA, PEDV/PDCoV, TGEV/PED/PoRVA, and PDCoV/PoRVA were 0.39 %, 0.11 %, 0.01 %, and 0.03 %, respectively, with no detection of other co-infections, as determined by the quadruplex real-time RT-qPCR. This research not only established a valuable tool for the simultaneous differentiation of TGEV, PEDV, PDCoV, and PoRVA in practical applications but also provided crucial insights into the prevalence of these viral pathogens causing diarrhea in Guangxi. • The quadruplex RT-qPCR method shows high specificity and efficiency, enabling simultaneous lab diagnosis of TGEV, PEDV, PDCoV, and PoRVA. It reduces detection costs by 75% and boosts efficiency by 300%. • The developed quadruplex RT-qPCR kit is comparable to the commercial kit from GM Biotechnology INT, implying its commercial potential. PEDV, PoRVA, and PEDV/PoRVA co-infections are the main causes of pig diarrhea in Guangxi, with TGEV and PDCoV detection rates being secondary factors during 2022-2023. [ABSTRACT FROM AUTHOR]