G9 Strains Research Articles

Development of a safe and broad-spectrum attenuated PEDV vaccine candidate by S2 subunit replacement.

Emerging highly virulent porcine epidemic diarrhea virus (PEDV) G2 strains has caused substantial economic losses worldwide. Vaccination with a live attenuated vaccine is a promising method to prevent and control PED because it can induce a strong immune response (including T- and B-cell immunity). Previous studies have demonstrated that the S2 subunit of the PEDV spike (S) protein is the determinant of PEDV trypsin independence. Here, we evaluated the pathogenicity, tissue tropism, and immunogenicity of the chimeric virus (YN-S2DR13) via animal experiments. We demonstrated that YN-S2DR13 strain, as a trypsin independent strain, increased intracellular proliferation capacity, significantly reduced virulence, and induced broad-spectrum neutralization protection against PEDV G1 and G2 strains. In vitro passaging data also validated the stability of YN-S2DR13. Our results showed that generating a chimeric PEDV strain that is trypsin-independent by replacing the S2 subunit is a promising approach for designing a live attenuated vaccine for PEDV in the future.

Genetic Characteristics of Echinococcus granulosus from Fixed Paraffin-Embedded Tissue Samples in Human Isolates Based on the High-Resolution Melting Point Analysis in Sabzevar, Northeast Iran.

There are ten genotypes of Echinococcus granulosus with different intermediate and final hosts affecting the parasite's life cycle and its transmission to humans. Therefore, this study was conducted to determine the genotype of isolated hydatid cysts using the simple and fast high-resolution melting point analysis (HRM) method. The paraffin tissue samples of patients who underwent surgery were obtained from the pathology sample bank of Vasei and Emdad Hospitals in Sabzevar, Iran during 2010-2020. The DNA content of the samples was extracted after collecting and determining the characteristics using the DNA extraction kit. PCR was performed on the samples and the presence of the hydatid cyst genome was confirmed using the special Master Kit. Mix PCR of Solis Biodyne Company and Real-Time device (Bio-Rad) were used, and the genetic identity of hydatid cysts were determined. Out of 33 paraffin samples, 21 samples contained hydatid cyst DNA, two of which were from the brain and 19 from the liver tissues; 12 samples did not contain hydatid cyst DNAs. All liver samples were from sheep species (G1), and the brain samples were from buffalo species (G3). Therefore, 9.53% of the Echinococcus species collected were buffalo (G3), and 90.47% were sheep (G1) strain. Based on previous patterns, HRM methods can be used for easy and quick identification of Echinococcus strains. The G1 strain was the dominant strain causing hydatid cyst in different human organs, including the liver and brain.

Evaluation of the Molecular Properties of Echinococcus granulosus Isolates from Various Hosts in Şanlıurfa Province

Cystic echinococcosis (CE), which is raised by the larvae of Echinococcus granulosus (E. granulosus), is single of the world's most significant zoonoses, affecting both humans and animals. In this labor, it was the goal to perform genotyping of hydatid cyst stocks collected from various regions of Şanlıurfa and diverse middle hosts by molecular methods. A total of 197 hydatid cyst samples were obtained, including 40 pre-diagnosed human samples with paraffin, Türkiye 140 sheep, and 17 cattle samples. After microscopic examination, 33 human isolates and 80 sheep cyst are regarded fertile because they contain more than 250 protoscolex and/or scolex hooks. PCR-RFLP technique was used to determine the genetics of fertile isolates by analyzing the ribosomal DNA internal transcribed spacer gene 1 (rDNA-ITS1) region of the samples. As a result of having the same band profile, it was found that all of the isolates were common sheep strain (G1). Echinococcus granulosus G1 strain was the reference material. This study demonstrated that the local sheep lineage of E. granulosus is the dominant genotype in Şanlıurfa province. It is thought to lead to further work on the epidemiology and ecology of parasites in animals in this region and neighboring countries.

Whole-Genome Characterization of Rotavirus G9P[6] and G9P[4] Strains That Emerged after Rotavirus Vaccine Introduction in Mozambique.

Mozambique introduced the Rotarix® vaccine into the National Immunization Program in September 2015. Following vaccine introduction, rotavirus A (RVA) genotypes, G9P[4] and G9P[6], were detected for the first time since rotavirus surveillance programs were implemented in the country. To understand the emergence of these strains, the whole genomes of 47 ELISA RVA positive strains detected between 2015 and 2018 were characterized using an Illumina MiSeq-based sequencing pipeline. Of the 29 G9 strains characterized, 14 exhibited a typical Wa-like genome constellation and 15 a DS-1-like genome constellation. Mostly, the G9P[4] and G9P[6] strains clustered consistently for most of the genome segments, except the G- and P-genotypes. For the G9 genotype, the strains formed three different conserved clades, separated by the P type (P[4], P[6] and P[8]), suggesting different origins for this genotype. Analysis of the VP6-encoding gene revealed that seven G9P[6] strains clustered close to antelope and bovine strains. A rare E6 NSP4 genotype was detected for strain RVA/Human-wt/MOZ/HCN1595/2017/G9P[4] and a genetically distinct lineage IV or OP354-like P[8] was identified for RVA/Human-wt/MOZ/HGJM0644/2015/G9P[8] strain. These results highlight the need for genomic surveillance of RVA strains detected in Mozambique and the importance of following a One Health approach to identify and characterize potential zoonotic strains causing acute gastroenteritis in Mozambican children.

Assessment of non-target toxicity of insecticides on Ganaspis brasiliensis (Ihering) in laboratory and field conditions.

G1 strain Ganaspis brasiliensis (Ihering) has been recently released in both Europe and America as a biological control agent of the spotted wing drosophila, Drosophila suzukii (Matsumura). In initial phases of classical biological control programs, it becomes imperative to evaluate the susceptibility of parasitoids to insecticides, to identify the best alternatives to adopt in an integrated pest management and organic perspective. In this study, we evaluated lethal and sublethal effects of topical application of five different insecticides classes: neonicotinoids, diamides, pyrethroids, organophosphates and spinosyns. Additionally, we tested residual toxicity in field trials in vineyards and sweet cherry orchards. Adult wasps' susceptibility to different insecticides' classes were consistent between laboratory and field. Spinosad exhibited the highest toxicity, with a median lethal concentration (LC50) of 0.00372 of the maximum field dose, and the highest knock-down effect in field trials, causing 92.5 ± 5% of mortality at T0. λ-cyhalothrin showed sublethal effects on both male and female insects' longevity when applied at LC30. In field trials, deltamethrin showed the highest persistence, causing significant parasitoid mortality up to 14 days after treatment. Conversely, cyantraniliprole was the least toxic active ingredient according to both topical and residual bioassays, even though its residues caused mortality up to 7 days after the treatment in the field. Our results indicate that spinosad and λ-cyhalothrin are highly toxic to G. brasiliensis, making them incompatible with classical biological control programs. Cyantraniliprole exhibited lower toxicity, and may be considered a selective pesticide for the integrated management of D. suzukii. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Molecular evolutionary analysis of novel NSP4 mono-reassortant G1P[8]-E2 rotavirus strains that caused a discontinuous epidemic in Japan in 2015 and 2018.

In the 2010s, several unusual rotavirus strains emerged, causing epidemics worldwide. This study reports a comprehensive molecular epidemiological study of rotaviruses in Japan based on full-genome analysis. From 2014 to 2019, a total of 489 rotavirus-positive stool specimens were identified, and the associated viral genomes were analyzed by next-generation sequencing. The genotype constellations of those strains were classified into nine patterns (G1P[8] (Wa), G1P[8]-E2, G1P[8] (DS-1), G2P[4] (DS-1), G3P[8] (Wa), G3P[8] (DS-1), G8P[8] (DS-1), G9P[8] (Wa), and G9P[8]-E2). The major prevalent genotype differed by year, comprising G8P[8] (DS-1) (37% of that year's isolates) in 2014, G1P[8] (DS-1) (65%) in 2015, G9P[8] (Wa) (72%) in 2016, G3P[8] (DS-1) (66%) in 2017, G1P[8]-E2 (53%) in 2018, and G9P[8] (Wa) (26%) in 2019. The G1P[8]-E2 strains (G1-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1) isolated from a total of 42 specimens in discontinuous years (2015 and 2018), which were the newly-emerged NSP4 mono-reassortant strains. Based on the results of the Bayesian evolutionary analyses, G1P[8]-E2 and G9P[8]-E2 were hypothesized to have been generated from distinct independent inter-genogroup reassortment events. The G1 strains detected in this study were classified into multiple clusters, depending on the year of detection. A comparison of the predicted amino acid sequences of the VP7 epitopes revealed that the G1 strains detected in different years encoded VP7 epitopes harboring distinct mutations. These mutations may be responsible for immune escape and annual changes in the prevalent strains.

Whole-genome analysis of human group A rotaviruses in 1980s Japan and evolutionary assessment of global Wa-like strains across half a century.

Historically, the Wa-like strains of human group A rotavirus (RVA) have been major causes of gastroenteritis. However, since the 2010s, the circulation of non-Wa-like strains has been increasingly reported, indicating a shift in the molecular epidemiology of RVA. Although understanding RVA evolution requires the analysis of both current and historical strains, comprehensive pre-1980's sequencing data are scarce globally. We determined the whole-genome sequences of representative strains from six RVA gastroenteritis outbreaks observed at an infant home in Sapporo, Japan, between 1981 and 1989. These outbreaks were mainly caused by G1 or G3 Wa-like strains, resembling strains from the United States in the 1970s-1980s and from Malawi in the 1990s. Phylogenetic analysis of these infant home strains, together with Wa-like strains collected worldwide from the 1970s to 2020, revealed a notable trend: pre-2010 strains diverged into multiple lineages in many genomic segments, whereas post-2010 strains tended to converge into a single lineage. However, Bayesian skyline plot indicated near-constant effective population sizes from the 1970s to 2020, and selection pressure analysis identified positive selection only at amino acid 75 of NSP2. These results suggest that evidence supporting the influence of rotavirus vaccines, introduced globally since 2006, on Wa-like RVA molecular evolution is lacking at present, and phylogenetic analysis may simply reflect natural fluctuations in RVA molecular evolution. Evaluating the long-term impact of RV vaccines on the molecular evolution of RVA requires sustained surveillance.

Exploring Variability: Inflammation Mediator Levels across Tissues and Time in Poultry Experimentally Infected by the G1a and G6 Genogroups of Infectious Bursal Disease Virus (IBDV).

Infectious bursal disease virus (IBDV) is a significant burden for poultry production and market due to both direct disease and induced immunosuppression. In the present study, the expression of different cytokines in the bursa of Fabricius and thymus was evaluated during a 28-day-long experimental infection with two strains classified in the G1a (Classical) and G6 (ITA) genogroups. Although both strains significantly affected and modulated the expression of different molecules, the G6 strain seemed to induce a delayed immune response or suppress it more promptly. A recovery in the expression of several mediators was observed in the G1a-infected group at the end of the study, but not in the G6 one, further supporting a more persistent immunosuppression. This evidence fits with the higher replication level previously reported for the G6 and with the clinical outcome, as this genotype, although subclinical, has often been considered more immunosuppressive. However, unlike other studies focused on shorter time periods after infection, the patterns observed in this paper were highly variable and complex, depending on the strain, tissue, and time point, and characterized by a non-negligible within-group variability. Besides confirming the strain/genogroup effect on immune system modulation, the present study suggests the usefulness of longer monitoring activities after experimental infection to better understand the complex patterns and interactions with the host response.

A comparative study of the biosynthesis of CuNPs by Niallia circulans G9 and Paenibacillus sp. S4c strains: characterization and application as antimicrobial agents

BackgroundBiosynthesis of metallic nanoparticles using microorganisms are a fabulous and emerging eco-friendly science with well-defined sizes, shapes and controlled monodispersity. Copper nanoparticles, among other metal particles, have sparked increased attention due to their applications in electronics, optics, catalysis, and antimicrobial agents.ResultsThis investigation explains the biosynthesis and characterization of copper nanoparticles from soil strains, Niallia circulans G9 and Paenibacillus sp. S4c by an eco-friendly method. The maximum reduction of copper ions and maximum synthesis CuNPs was provided by these strains. Biogenic formation of CuNPs have been characterized by UV–visible absorption spectroscopy, X-ray diffraction, Fourier transform infrared spectroscopy, X-ray analysis and transmission electron microscopy analysis. Using UV-visible spectrum scanning, the synthesised CuNPs’ SPR spectra showed maximum absorption peaks at λ304&308 nm. TEM investigation of the produced CuNPs revealed the development of spherical/hexagonal nanoparticles with a size range of 13–100 nm by the G9 strain and spherical nanoparticles with a size range of 5–40 nm by the S4c strain. Functional groups and chemical composition of CuONPs were also confirmed. The antimicrobial activity of the biosynthesized CuNPs were investigated against some human pathogens. CuNPs produced from the G9 strain had the highest activity against Candida albicans ATCC 10,231 and the lowest against Pseudomonas aeruginosa ATCC 9027. CuNPs from the S4c strain demonstrated the highest activity against Escherichia coli ATCC 10,231 and the lowest activity against Klebsiella pneumonia ATCC 13,883.ConclusionThe present work focused on increasing the CuNPs production by two isolates, Niallia circulans G9 and Paenibacillus sp. S4c, which were then characterized alongside. The used analytics and chemical composition techniques validated the existence of CuONPs in the G9 and S4c biosynthesized nano cupper. CuNPs of S4c are smaller and have a more varied shape than those of G9 strain, according to TEM images. In terms of antibacterial activity, the biosynthesized CuNPs from G9 and S4c were found to be more effective against Candida albicans ATCC 10,231 and E. coli ATCC 10,231, respectively.

Non-target effects of neurotoxic insecticides on Ganaspis cf. brasiliensis, a classical biological control agent of the spotted wing Drosophila

BackgroundThe spotted wing drosophila, Drosophila suzukii, is an invasive pest causing significant economic losses worldwide. Current pest control strategies mainly rely on insecticides, which negatively impact fruit marketability and the sustainability of integrated pest management (IPM) programs. In addition, pesticides can have dramatic consequences on non-target species when persisting in the environment at low concentrations after field applications. In this context, chemical control can strongly interfere with the releases of the G1 strain of the Asian larval parasitoid Ganaspis cf. brasiliensis, which is currently the adopted classical biological control agent to manage D. suzukii infestations worldwide.MethodsProbit analysis was used to assess the baseline toxicity of acetamiprid, cyazypyr, lambda-cyhalothrin, phosmet, and spinosad on G1 G. cf. brasiliensis adults through residual contact exposure in the laboratory. Then, adult parasitoids were exposed to insecticide low Lethal Concentrations (LC5 and LC30) and their mortality was checked daily to assess the survival of treated wasps.ResultsLambda-cyhalothrin showed the highest toxicity on the parasitoid with a LC50 of 1.38 × 10–3 g active ingredient (a.i.) /L, while cyazypyr seemed the safer active ingredient with an estimated LC50 of 0.20 g a.i./L without affecting parasitoids at sublethal doses. Spinosad and phosmet significantly reduced wasp survival at both LC30 and LC5, while lambda-cyhalothrin and acetamiprid affected parasitoid lifespan only at LC30. Spinosad, lambda-cyhalothrin and phosmet LC30 caused the major survival reductions, followed by acetamiprid LC30. The least significant reduction in parasitoid survival was 21.6% by spinosad LC5.ConclusionsOverall, this study highlighted the importance of carefully selecting insecticides to minimize adverse effects on non-target organisms. In particular, cyazypyr was the most promising candidate to integrate inoculative biological control with chemical treatments. By contrast, the application of phosmet, spinosad and lambda-cyhalothrin should be avoided alongside parasitoid field releases. Although acetamiprid is less used against D. suzukii in the field than the other tested molecules, it should be used with caution due to its sublethal toxicity on the parasitoid. These results provide the first evidence of G. cf. brasiliensis susceptibility to insecticides in order to promote sustainable and efficient pest management strategies.

Genetic diversity and evolution of G12P[6] DS-1-like and G12P[9] AU-1-like Rotavirus strains in Brazil.

In the early 2000s, the global emergence of rotavirus (RVA) G12P[8] genotype was noted, while G12P[6] and G12P[9] combinations remained rare in humans. This study aimed to characterize and phylogenetically analyze three Brazilian G12P[9] and four G12P[6] RVA strains from 2011 to 2020, through RT-PCR and sequencing, in order to enhance our understanding of the genetic relationship between human and animal-origin RVA strains. G12P[6] strains displayed a DS-1-like backbone, showing a distinct genetic clustering. G12P[6] IAL-R52/2020, IAL-R95/2020 and IAL-R465/2019 strains clustered with 2019 Northeastern G12P[6] Brazilian strains and a 2018 Benin strain, whereas IAL-R86/2011 strain grouped with 2010 Northern G12P[6] Brazilian strains and G2P[4] strains from the United States and Belgium. These findings suggest an African genetic ancestry and reassortments with co-circulating American strains sharing the same DS-1-like constellation. No recent zoonotic reassortment was observed, and the DS-1-like constellation detected in Brazilian G12P[6] strains does not seem to be genetically linked to globally reported intergenogroup G1/G3/G9/G8P[8] DS-1-like human strains. G12P[9] strains exhibited an AU-1-like backbone with two different genotype-lineage constellations: IAL-R566/2011 and IAL-R1151/2012 belonged to a VP3/M3.V Lineage, and IAL-R870/2013 to a VP3/M3.II Lineage, suggesting two co-circulating strains in Brazil. This genetic diversity is not observed elsewhere, and the VP3/M3.II Lineage in G12P[9] strains seems to be exclusive to Brazil, indicating its evolution within the country. All three G12P[9] AU-1-like strains were closely relate to G12P[9] strains from Paraguay (2006-2007) and Brazil (2010). Phylogenetic analysis also highlighted that all South American G12P[9] AU-1-like strains had a common origin and supports the hypothesis of their importation from Asia, with no recent introduction from globally circulating G12P[9] strains or reassortments with local G12 strains P[8] or P[6]. Notably, certain genes in the Brazilian G12P[9] AU-1-like strains share ancestry with feline/canine RVAs (VP3/M3.II, NSP4/E3.IV and NSP2/N3.II), whereas NSP1/A3.VI likely originated from artiodactyls, suggesting a history of zoonotic transmission with human strains. This genomic data adds understanding to the molecular epidemiology of G12P[6] and G12P[9] RVA strains in Brazil, offering insights into their genetic diversity and evolution.

Antigenic epitope analysis of Pakistani G3 and G9 rotavirus strains compared to vaccine strains revealed multiple amino acid differences

Rotaviruses belong to genotype VP4-P[8] are a significant cause of severe loose diarrhea in infants and young children. In the present study, we characterised the complete genome of three of the Pakistani P[8]b RVA strains by Illumina HiSeq sequencing technology to determine the complete genotype constellation providing insight into the evolutionary dynamics of their genes using maximum likelihood analysis. The maximum genomic sequences of our study strains were similar to more recent human Wa-Like G1P[8]a, G3P[8]a, G4P[6], G4P[8], G9P[4], G9P[8]a, G11P[25],G12P[8]a and G12P[6] strains circulating around the world. Therefore, strains PAK274, PAK439 and PAK624 carry natively distinctive VP4 gene with universally common human Wa-Like genetic backbone. Comparing our study P[8]b strains with vaccines strains RotarixTM and RotaTeqTM, multiple amino acid differences were examined between vaccine virus antigenic epitopes and Pakistani isolates. Over time, these differences may result in the selection for strains that will escape the vaccine-induced RVA-neutralizing-antibody effect.

Complete characterization of NADH dehydrogenase subunit 1 gene in human hydatid cysts

يمتلك طفيلي الاكياس العدرية سلالاتين متشابهتين (G1 و G3) ونظرا للدور البارز الذي تلعبه عائلة جينات نازعات الهيدروجين في التميز بين هاتين السلالاتين، لذا هدفت الدراسة الحالية الى اجراء التوصيف الجزيئي الكامل لاحد ابرز انواع هذه العائلة وهو الجين NAD1 (من شفرة البدء الى الي شفرة التوقف) للاكياس العدرية المعزولة جراحيا من اكباد المرضى (ولمحافظتي البصرة وميسان) في مستشفى الصدر التعليمي في محافظة البصرة، العراق. عزلت الرؤيسات الاولية من الاكياس تحت ظروف معقمة واستخدمت في استخلاص الحامض النووي منقوص الاوكسجين. اذ ضخم التتابع الكامل للجين في عينات الدراسة وانجز التتابع (السكونس) لهما باستخدام بادئات متخصصه. اظهرت نتائج تحليل السكونس المتكرر Consensus sequence امتلاك كل من عينات البصرة و ميسان 1038 زوجا من القواعد وتقع شفرة البدء GTG في الموقع 87-89 وشفرة التوقف TAA في الموقع 978-980. كما اظهرت استدلالات الاحماض الامينية الى وجود 297 حامض اميني لبروتينات عينات البصرة وميسان. فضلا عن ذلك بينت تحاليل المعلومات الحياتية احتواء هذه البروتينات على الحامض الاميني Isoleucine في الموقع 188 (I188) وهو موجود فقط في سلالة الاغنام G1. وان النموذج التركيبي لبروتينات عينات البصرة وميسان اظهر تشابها كبيرا و يقع I188 في الحلزون 10 (α10). بينت الدراسة ولاول مرة على مستوى العراق التوصيف الكامل للجين NAD1 والعائد الى سلالة الاغنام G1 المسببه لمرض الاكياس العدرية البشري.

Immunological properties of inactivated vaccines against Newcastle disease based on virus antigens VII and II the genotype

Newcastle disease (ND) is now widely spread in many countries of Eurasia, Africa and the Americas and causes significant economic damage to poultry farming. New NDinfected localities are reported in the Russian Federation every year; besides, the number of farms with vaccinated poultry demonstrating high specific antibody titres, decreased performance and survivability while being asymptomatic has increased. This is attributed to the virus evolution leading to the emergence of new genotypes represented by highly virulent strains. Since 2019, subgenotype VII-L has been isolated from Newcastle disease outbreaks in Russia. The vaccines currently available on the market do not confer appropriate protection against it due to the lack of antigenic matching between vaccine strains and this subgenotype. The development of a vaccine based on subgenotype VII -L will allow to ensure protection of poultry flocks against Newcastle disease caused by new NDV isolates. The immunological properties of emulsion inactivated vaccine against Newcastle disease based on genotype VII NDV (ARRIAH G7 strain) antigen were studied in 30-day-old Hisex Brown chickens. A similar vaccine based on genotype II NDV LaSota strain antigen was used as control. Tests of different immunizing antigen doses were carried out. The vaccines were tested for induced humoral immunity level (HI antibody titres), as well as for protective effect (protective index) after challenge with a highly virulent genotype VII NDV strain. The tests revealed that the antigenic activity and protective effect of both vaccines depended on the antigen concentration in the inoculation volume. Antibody titres were always higher when homologous antigens were used in HI tests. The vaccine based on the antigen of ARRIAH G7 strain (homologous to the challenge strain) was more effective than the control vaccine. Less ARRIAH G7 strain antigen was required to protect 50% of vaccinated poultry, compared with LaSota strain antigen. The comparative assessment of the same doses of the antigens showed that the vaccine based on ARRIAH G7 strain antigen had better immunological properties. The tested antigens demonstrated 18.3% bilateral affinity.

Genetic diversity of G9, G3, G8 and G1 rotavirus group A strains circulating among children with acute gastroenteritis in Vietnam from 2016 to 2021

Rotavirus group A (RVA) is the most common cause of severe childhood diarrhea worldwide. The introduction of rotavirus vaccination programs has contributed to a reduction in hospitalizations and mortality caused by RVA. From 2016 to 2021, we conducted surveillance to monitor RVA prevalence and genotype distribution in Nam Dinh and Thua Thien Hue (TT Hue) provinces where a pilot Rotavin-M1 vaccine (Vietnam) implementation took place from 2017 to 2020. Out of 6626 stool samples, RVA was detected in 2164 (32.6%) by ELISA. RT-PCR using type-specific primers were used to determine the G and P genotypes of RVA-positive specimens. Whole genome sequences of a subset of 52 specimens randomly selected from 2016 to 2021 were mapped using next-generation sequencing. From 2016 to 2021, the G9, G3 and G8 strains dominated, with detected frequencies of 39%, 23%, and 19%, respectively; of which, the most common genotypes identified were G9P[8], G3P[8] and G8P[8]. G1 strains re-emerged in Nam Dinh and TT Hue (29.5% and 11.9%, respectively) from 2020 to 2021. G3 prevalence decreased from 74% to 20% in TT Hue and from 21% to 13% in Nam Dinh province between 2017 and 2021. The G3 strains consisted of 52% human typical G3 (hG3) and 47% equine-like G3 (eG3). Full genome analysis showed substantial diversity among the circulating G3 strains with different backgrounds relating to equine and feline viruses. G9 prevalence decreased sharply from 2016 to 2021 in both provinces. G8 strains peaked during 2019–2020 in Nam Dinh and TT Hue provinces (68% and 46%, respectively). Most G8 and G9 strains had no genetic differences over the surveillance period with very high nucleotide similarities of 99.2–99.9% and 99.1–99.7%, respectively. The G1 strains were not derived from the RVA vaccine. Changes in the genotype distribution and substantial diversity among circulating strains were detected throughout the surveillance period and differed between the two provinces. Determining vaccine effectiveness against circulating strains over time will be important to ensure that observed changes are due to natural secular variation and not from vaccine pressure.

China-origin G1 group isolate FPV072 exhibits higher infectivity and pathogenicity than G2 group isolate FPV027.

Feline parvovirus (FPV), a single-stranded DNA virus, is accountable for causing feline panleukopenia, a highly contagious and often lethal disease that primarily affects cats. The epidemiology prevalence and pathogenicity of FPV in certain regions of China, however, remains unclear. The aim of this research was to investigate the epidemiology of FPV in different regions of China in 2021 and compare its infectivity and pathogenicity. In this research, a total of 36 FPV strains were obtained from diverse regions across China. Phylogenetic analysis was performed based on the VP2 and NS1 sequences, and two representative strains, FPV027 and FPV072, which belonged to different branches, were selected for comparative assessment of infectivity and pathogenicity. The results revealed that all strains were phylogenetically classified into two groups, G1 and G2, with a higher prevalence of G1 strains in China. Both in vitro and in vivo experiments demonstrated that FPV072 (G1 group) exhibited enhanced infectivity and pathogenicity compared to FPV027 (G2 Group). The structural alignment of the VP2 protein between the two viruses revealed mutations in residues 91, 232, and 300 that may contribute to differences in infectivity and pathogenicity. The findings from these observations will contribute significantly to the overall understanding of the molecular epidemiology of FPV in China and facilitate the development of an effective FPV vaccine.

Emergence and high prevalence of unusual rotavirus G8P[8] strains in outpatients with acute gastroenteritis in Shanghai, China.

Group A rotavirus (RVA) is considered an important cause of acute gastroenteritis (AGE) in all age groups, especially in children. We investigated the epidemiology of RVA in outpatients aged ≤ 16 years at the Children's Hospital of Fudan University, Shanghai, China. In this study, 16.6% (246/1482) were infected with RVA. The detection rate of RVA was significantly higher in the year of 2021 (20.3%, 147/725) compared to the year of 2020 (14.5%, 77/531) and 2022 (9.7%, 22/226) (p = 0.000). RVA infection was prevalent in all seasons from 2020 to 2022, with a different monthly distribution observed in different years. Among 246 RVA-positive samples, 14 different RVA genotypes were detected with different frequencies. Overall, G9P[8] (45.5%, 112/246) was the most common RVA genotype, followed by G8P[8] (37.4%, 92/246) and G3P[8] (4.1%, 10/246). The prevalence of G/P combinations varied from 2020 to 2022. G9P[8] was the most prevalent circulating genotype in 2020 (68.2%, 15/22) and 2021 (57.8%, 85/147). However, G8P[8] (68.8%, 53/77) suddenly became the most prevalent genotype in 2022 after being first identified in 2020 and prevalent in 2021. The G8 strains detected in the study were all clustered to DS-1-like G8 strains with the closest genetic distance to strains circulating in Southeast Asia. Our study demonstrated the diversity of circulating RVA genotypes in Shanghai. The sudden emergence and high prevalence of unusual G8P[8] strains deserve more concern and indicate the need for continuous surveillance of RVA in children with AGE in the future to refine future vaccine strategy.

Emergence of Intergenogroup Reassortant G9P[4] Strains Following Rotavirus Vaccine Introduction in Ghana.

Rotavirus (RVA) is a leading cause of childhood gastroenteritis. RVA vaccines have reduced the global disease burden; however, the emergence of intergenogroup reassortant strains is a growing concern. During surveillance in Ghana, we observed the emergence of G9P[4] RVA strains in the fourth year after RVA vaccine introduction. To investigate whether Ghanaian G9P[4] strains also exhibited the DS-1-like backbone, as seen in reassortant G1/G3/G8/G9 strains found in other countries in recent years, this study determined the whole genome sequences of fifteen G9P[4] and two G2P[4] RVA strains detected during 2015-2016. The results reveal that the Ghanaian G9P[4] strains exhibited a double-reassortant genotype, with G9-VP7 and E6-NSP4 genes on a DS-1-like backbone (G9-P[4]-I2-R2-C2-M2-A2-N2-T2-E6-H2). Although they shared a common ancestor with G9P[4] DS-1-like strains from other countries, further intra-reassortment events were observed among the original G9P[4] and co-circulating strains in Ghana. In the post-vaccine era, there were significant changes in the distribution of RVA genotype constellations, with unique strains emerging, indicating an impact beyond natural cyclical fluctuations. However, reassortant strains may exhibit instability and have a limited duration of appearance. Current vaccines have shown efficacy against DS-1-like strains; however, ongoing surveillance in fully vaccinated children is crucial for addressing concerns about long-term effectiveness.

Echinococcus granulosus sheep strain (G1) as the predominant genotype in definitive host (dogs) isolates in northeastern Iran

Echinococcus granulosus sensu lato (E. granulosus s.l.) is a zoonotic parasite, causing cystic echinococcosis in humans. In the present study, prevalence and genotypes of E. granulosus s.l. was assessed in stools collected from 244 dogs including 138 stray and 106 domestic animals using high resolution melting curve (HRM) method. Initially, to detect taeniid eggs in feces, all samples were examined using the formalin-ether techniques. Genomic DNA was extracted from the positive samples and E. granulosus s.l. was differentiated from other Taeniidae parasites using SSU-rDNA gene and E. granulosus s.l. was analyzed for genotyping using HRM based on the cox1 gene. In total, 12.7% (31/244) of the samples were positive for Taeniidae eggs. In addition, among the positive samples, 77.4% (24/31) were positive for E. granulosus s.l.. In details, 11.3% (12/106) of the domestic dogs and 8.7% (12/138) of the stray dogs were positive for E. granulosus s.l.. The results of HRM analysis showed that all E. granulosus s.l. isolates were G1 strain. Findings of the present study indicated a considerable prevalence of E. granulosus G1 among dogs in the northeast of Iran and imply a serious risk of transmitting to humans and livestock.

Genomic Analysis of Rwandan G9P[8] Rotavirus Strains Pre- and Post-RotaTeq® Vaccine Reveals Significant Distinct Sub-Clustering in a Post-Vaccination Cohort.

Although the introduction of rotavirus vaccines has substantially contributed to the reduction in rotavirus morbidity and mortality, concerns persist about the re-emergence of variant strains that might alter vaccine effectiveness in the long term. The G9 strains re-emerged in Africa during the mid-1990s and have more recently become predominant in some countries, such as Ghana and Zambia. In Rwanda, during the 2011 to 2015 routine surveillance period, G9P[8] persisted during both the pre- and post-vaccine periods. The pre-vaccination cohort was based on the surveillance period of 2011 to 2012, and the post-vaccination cohort was based on the period of 2013 to 2015, excluding 2014. The RotaTeq® vaccine that was first introduced in Rwanda in 2012 is genotypically heterologous to Viral Protein 7 (VP7) G9. This study elucidated the whole genome of Rwandan G9P[8] rotavirus strains pre- and post-RotaTeq® vaccine introduction. Fecal samples from Rwandan children under the age of five years (pre-vaccine n = 23; post-vaccine n = 7), conventionally genotyped and identified as G9P[8], were included. Whole-genome sequencing was then performed using the Illumina® MiSeq platform. Phylogenetic analysis and pair-wise sequence analysis were performed using MEGA6 software. Distinct clustering of three post-vaccination study strains was observed in all 11 gene segments, compared to the other Rwandan G9P[8] study strains. Specific amino acid differences were identified across the gene segments of these three 2015 post-vaccine strains. Important amino acid differences were identified at position N242S in the VP7 genome segment of the three post-vaccine G9 strains compared to the other G9 strains. This substitution occurs at a neutralization epitope site and may slightly affect protein interaction at that position. These findings indicate that the Rwandan G9P[8] strains revealed a distinct sub-clustering pattern among post-vaccination study strains circulating in Rwanda, with changes at neutralization epitopes, which may play a role in neutralization escape from vaccine candidates. This emphasizes the need for continuous whole-genome surveillance to better understand the evolution and epidemiology of the G9P[8] strains post-vaccination.