G9 Strains Research Articles

The origins of G12P[6] rotavirus strains detected in Lebanon.

The G12 rotaviruses are an increasingly important cause of severe diarrhoea in infants and young children worldwide. Seven human G12P[6] rotavirus strains were detected in stool samples from children hospitalized with gastroenteritis in Lebanon during a 2011-2013 surveillance study. Complete genomes of these strains were sequenced using VirCapSeq-VERT, a capture-based high-throughput viral-sequencing method, and further characterized based on phylogenetic analyses with global RVA and vaccine strains. Based on the complete genomic analysis, all Lebanese G12 strains were found to have Wa-like genetic backbone G12-P[6]-I1-R1-C1-M1-A1-N1-T1-E1-H1. Phylogenetically, these strains fell into two clusters where one of them might have emerged from Southeast Asian strains and the second one seems to have a mixed backbone between North American and Southeast Asian strains. Further analysis of these strains revealed high antigenic variability compared to available vaccine strains. To our knowledge, this is the first report on the complete genome-based characterization of G12P[6] emerging in Lebanon. Additional studies will provide important insights into the evolutionary dynamics of G12 rotaviruses spreading in Asia.

Isolation, characterisation and continuous culture of Lactobacillus spp. and its potential use for lactic acid production from whey

Bioprospecting of microorganisms with the potential of exploiting agro-industrial wastes is an active field of biotechnological research. In this study, we aimed to isolate a microorganism adapted to the dairy matrix with specific characteristics to produce lactic acid from dairy industry wastes, such as whey. Whey was collected from the agricultural region of Antioquia, Colombia, and samples were cultured on Man–Rogosa–Sharpe modified medium using lactose as a source of carbon. The most promising strain isolated was characterised, and its adaptation to whey was determined. A gram-positive Lactobacillus was isolated and named as G1. This strain grows in deproteinised whey with low protein requirements, has a homofermentative metabolism and exhibits high tolerance to low pH (≤5.0). The G1 strain is a promising bacterium for use in lactic acid production from whey, which is an underutilised by-product of the dairy industry.

Decade-long temporal analyses of circulating rotavirus genotypes during 2008–2017 in Eastern India: Phylodynamics during the pre-vaccination scenario

Background: Group-A human rotaviruses (GARV) are among the leading etiological agents causing acute childhood gastroenteritis, worldwide. Despite the significant reduction in global infantile death toll due to rotaviral diarrhea, developing countries like India still contributes substantially to rotavirus-related hospitalization and mortality rates. In on-going hospital based diarrheal-disease surveillance in Kolkata, eastern India (2008–17), GARV was identified as the most common cause of infantile gastroenteritis. The circulating strains were genotyped and phylogenetically analysed to understand their dynamics and evolution prior to the vaccine introduction in eastern India. Methods and materials: Stool samples were screened from children (≤5 years) with diarrhea, seeking health care facilities at two hospitals in Kolkata. Preliminary screening for rotavirus VP6 antigen was done by ELISA. GARV positive samples were genotyped by multiplex semi-nested PCR. DNA sequencing of VP7 (G-type) and VP4 (P-type) gene segments were done followed by phylogenetic analyses by MEGA 7 software. Results: Among 7092 stool samples, 3020 (42.5%) were found to be GARV positive. Children of 6–12 months age group were found most susceptible to infection (57.3%). GARV incidence peaked during the winter months each year (November–February). G1, G2, G3, G9 and G12 in conjunction with P[4], P[6] and P[8] genotypes were seen to co-circulate in the study-population. G1 strains predominated during 2008–10, while G2 and G9 genotypes eventually upsurged during 2011–13. G1 re-established its lead during 2013–15, while polymorphic G3 strains emerged for the first time in eastern India in 2015 and rooted itself as the cardinal strain during 2016–17. All the strains clustered distantly from the vaccine strains in the phylogenetic dendrograms, depicting less genetic relatedness. Differences in amino-acid residues encompassing the antigenic epitopes of VP4 and VP7 capsid proteins between the vaccine and Kolkata strains were observed. Conclusion: GARV shows a tendency towards directional selection under persistent immunological pressure that might promote natural selection of one genotype over another in an endemic setting. These decade-long temporal data during the pre-vaccination period revealed high hospitalization rates, along with dynamic circulation of multiple genotypes in addition to sporadic occurrence of zoonotic strains like G10P[6] and G11P[25].

Post Vaccination Epidemiology and Genotyping of Rotavirus Gastroenteritis at a Tertiary Care Centre of North-East Rajasthan.

To estimate the proportion of rotavirus diarrhea among hospitalized children aged under-five years, to determine the circulating rotavirus genotypes and to know impact rotavirus vaccine on prevalence and severity of rotavirus diarrhea. This study was a hospital based cross-sectional observational study conducted over a period of 29 mo (September 2017 through January 2020). Stool samples were collected from children who fall within the age range of 0-59 mo with acute diarrhea attending emergency or needing admission. Stool samples were tested for rotavirus by the enzyme linked immune-sorbent assay (ELISA) and genotyped using published methods. Out of 1480 samples, 360 (24.32%) cases were positive for rotavirus by ELISA, majority of them were male (62.97%). Maximum rotavirus positivity was found in the age group of <11 mo (55.27%). Statistically significance difference was seen in episodes of diarrhea and experience of vomiting in rotavirus diarrhea cases. Highest prevalence has been seen during winter season. The most prevalent G and P type combinations were G3P [8] strains [122 (34.08%)], G2P [4] [83 (23.18%)], G1P [8] [27 (7.54%)] and G9P [4] [20 (5.59%)]. Mixed strains contribute a significant proportion of stool sample. Rotavirus is an important cause of diarrhea in hospitalized children. There is continued circulation of G9 and G12 strains and the emergence of G3P [8] as most common strain.

Genotyping of the Echinococcus granulosus in Paraffin-Embedded Human Tissue Samples from Iran.

Cystic Echinococcosis (CE) is a medically important disease that is caused by the metacestodes of Echinococcus granulosus. Human hydatid is considered an endemic disease in specific regions of Iran. The goal of the present study was to determine the genetic diversity of E. granulosus from the paraffin-embedded human tissue samples which were collected from the endemic regions of Iran. Fifty-five formalin-fixed and paraffin-embedded hydatid cysts (FFPE) of humans, which had been removed surgically, were obtained from the South Khorasan and Sistan and Baluchistan provinces. These regions are related to the East and Southeast regions of Iran, respectively. The cox1 and nad1 genes from mitochondria were amplified from the extracted DNA and sequenced. The sequences were edited using the BioEdit software. Furthermore, phylogenetic and genetic diversity analyses were performed. Sequencing of the cox1 and nad1 genes from the 44 CE samples was done successfully. Genetic analysis revealed that 38 (86.3%) and 6 (13.6%) of the isolates were G1- and G6-genotypes, respectively. In general, eight and six haplotypes were identified by cox1 and nad1 genes analysis, respectively. For G1 strains, the haplotype diversity index was higher for the cox1 gene (0.6 ± 0.07) in comparison with the nad1 gene (0.4 ± 0.09). The findings of the present study showed that the sheep strain (G1) and the less important camel strain (G6) play the main roles in the transmission cycle of CE in the East and Southeast regions of Iran. Therefore, these results could be useful for managing the hydatid disease control programs in the studied and other similar areas.

Comparative genomics of the sequential Pseudomonas aeruginosa isolates obtained from the continuous imipenem stress evolution.

Pseudomonas aeruginosa is a major opportunistic human pathogen that causes nosocomial infections, and the proportion of carbapenem resistance has recently dramatically increased in P. aeruginosa due to the overuse of them. In this study, strains G10 and G20, with minimum inhibitory concentration (MIC) of imipenem of 16μg/ml and more than 32μg/ml, were isolated during continuous subculture of cells exposed to stepwise increasing concentrations of imipenem, respectively. The genomes of G10 and G20 were sequenced and compared with parental strain (P. aeruginosa ATCC 27853, G0). There were 59, 59, and 58 genes involved in antibiotic resistance which were predicted in G0, G10, and G20, respectively, while 374, 366, and 363 genes involved in virulence factors were identified among these three strains. Due to the significantly different MICs of imipenem and highly similar profiles of antibiotic resistance and virulence factors related genes among three strains, the specific genetic variations that occurred were identified and compared, including single nucleotide polymorphisms (SNPs), insertions and deletions (InDels), and structural variations (SVs). The increase in the MIC of imipenem was proposed to be linked to mutations involved in polyamine biosynthesis, biofilm formation, OprD, and efflux pump functions. This study aims to clarify the underlying mechanism of imipenem resistance and provide alternative strategies for reducing resistance in P. aeruginosa. KEY POINTS: • Strains with different imipenem MIC were obtained via laboratory selection evolution. • Whole genomes of two strains with different MIC of imipenem were sequenced. • Underlying mechanism of imipenem resistance was clarified via comparative genomics.

Occurrence and genetic diversity of rotavirus A in faeces of diarrheic calves submitted to a veterinary laboratory in Spain

A total of 237 faecal specimens from diarrheic calves younger than two months were collected and submitted for diagnosis of enteropathogens over a two-year period (2017–2018) to a veterinary laboratory. Samples originated from 193 dairy and beef farms in 29 provinces distributed throughout Spain, and were tested for the occurrence of three target enteric pathogens by reverse transcription real-time PCR (RT-qPCR): bovine rotavirus A (RVA), Cryptosporidium parvum and bovine coronavirus (BCoV). RT-PCR and nucleotide sequencing analysis were used to determine the G (VP7 gene) and P (VP4 gene) genotypes of 26 specimens positive for RVA. A total of 188 specimens (79.3 %) were positive for at least one of the three target enteric pathogens, and 101 samples (42.6 %) harbored mixed infections. The individual prevalence was 57.8 %, 50.6 % and 23.6 % for C. parvum, RVA and BCoV, respectively. Molecular analysis of selected RVA strains revealed the presence of the G6, G10, G3, P[5] and P[11] genotypes, with the combinations G6P[5] and G6P[11] being the most prevalent. Alignments of nucleotide sequences of the VP7 and VP4 markers showed a high frequency of single nucleotide polymorphisms (SNPs), with up to 294 SNPs found in 869bp of sequence at the G6 genotype (0.338 SNPs/nt), which reveals the extensive genetic diversity of RVA strains. Phylogenetic analysis of the VP7 gene of the G6 strains revealed four distinct lineages, with most strains clustering in the G6-IV lineage. The discrepancies between the RVA genotypes circulating in the sampled cattle farms and the genotypes contained in commercial vaccines currently available in Spain are discussed. We believe that this is the first study on the molecular characterization of rotavirus infecting cattle in Spain.

Genotyping and phylogenetic analysis of hydatid cysts isolated from livestock in Bushehr province, Iran.

Hydatid cyst is one of the parasitic zoonoses caused by infection with the larval stage of Echinococcus granulosus tapeworm. The spread of this parasite is global and is of great importance in terms of public health. To date, ten different species of this parasite have been identified that differ in characteristics such as life cycle, epidemiology and pathogenesis. The purpose of this study was to determine the genotype and phylogenetic relationship of hydatid cysts isolated from livestock of Bushehr province, Iran. About 62 samples of hepatic and pulmonary hydatid cysts were collected from slaughtered animals. DNA extracted by phenol-chloroform method was amplified by PCR using primers specific for the cox1 gene. The PCR products of 50 samples were sequenced and analyzed using BioEdit software and compared with sequences in the GenBank. The phylogenetic tree was drawn using Neighbor Joining tree-NJ method, and its reliability was evaluated. Sequencing results showed that out of 50 sequenced samples, 43 samples had the genotype of Echinococcus granulosus and 7 samples had the genotype of Taenia hydatigena. By drawing a phylogenetic tree, all 43 hydatid cyst samples belonged to G1 strain. The predominance of G1 strain of hydatid cyst in livestock of Bushehr province shows the main role of this genotype in establishing the life cycle of parasite in this region and if the genotype of the parasite in dogs and humans is determined, then these findings can be used to disrupt the life cycle of the parasite and reduce the human infections.

Whole Genome In-Silico Analysis of South African G1P[8] Rotavirus Strains Before and After Vaccine Introduction Over A Period of 14 Years.

Rotavirus G1P[8] strains account for more than half of the group A rotavirus (RVA) infections in children under five years of age, globally. A total of 103 stool samples previously characterized as G1P[8] and collected seven years before and seven years after introducing the Rotarix® vaccine in South Africa were processed for whole-genome sequencing. All the strains analyzed had a Wa-like constellation (G1-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1). South African pre- and post-vaccine G1 strains were clustered in G1 lineage-I and II while the majority (84.2%) of the P[8] strains were grouped in P[8] lineage-III. Several amino acid sites across ten gene segments with the exception of VP7 were under positive selective pressure. Except for the N147D substitution in the antigenic site of eight post-vaccine G1 strains when compared to both Rotarix® and pre-vaccine strains, most of the amino acid substitutions in the antigenic regions of post-vaccine G1P[8] strains were already present during the pre-vaccine period. Therefore, Rotarix® did not appear to have an impact on the amino acid differences in the antigenic regions of South African post-vaccine G1P[8] strains. However, continued whole-genome surveillance of RVA strains to decipher genetic changes in the post-vaccine period remains imperative.

Growth and production of sixth generation of brown rice mutants in a high altitude location

This paper presents the result of an experiment which aimed to determine the growth and production of some 6th generation Sinjai brown rice mutants (M6) in the highlands. The experiment was conducted in an irrigated paddy field with a height of 750 meters above sea level. The arrangement was a randomized block design (RBD). The treatments consisted of 8 brown rice mutant lines (g1, g2, g3, g4, g5, g6, g7, g8) and Sinjai brown rice parent as control (g9). The results showed that the mutant lines g2 and g3 produced the highest grain production of 3.10 tons ha−1 and 3.13 tons ha−1 respectively. Mutant strains g2 and g3 were short in plant height with a high number of productive tillers, flowering age and a faster harvest time, as well as highest grain length. g2 and g3 strains are the most potential lines to be developed in the highlands. The growth character and production components that have a real positive correlation to production per hectare are production per clump and presentation of filled grain per panicle.

The Presence of the Hairy-Root-Disease-Inducing (Ri) Plasmid in Wheat Endophytic Rhizobia Explains a Pathogen Reservoir Function of Healthy Resistant Plants.

A large number of strains in the Rhizobium radiobacter species complex (biovar 1 Agrobacterium) have been known as causative pathogens for crown gall and hairy root diseases. Strains within this complex were also found as endophytes in many plant species with no symptoms. The aim of this study was to reveal the endophyte variation of this complex and how these endophytic strains differ from pathogenic strains. In this study, we devised a simple but effective screening method by exploiting the high resolution power of mass spectrometry. We screened endophyte isolates from young wheat and barley plants, which are resistant to the diseases, and identified seven isolates from wheat as members of the R. radiobacter species complex. Through further analyses, we assigned five strains to the genomovar (genomic group) G1 and two strains to G7 in R. radiobacter Notably, these two genomovar groups harbor many known pathogenic strains. In fact, the two G7 endophyte strains showed pathogenicity on tobacco, as well as the virulence prerequisites, including a 200-kbp Ri plasmid. All five G1 strains possessed a 500-kbp plasmid, which is present in well-known crown gall pathogens. These data strongly suggest that healthy wheat plants are reservoirs for pathogenic strains of R. radiobacterIMPORTANCE Crown gall and hairy root diseases exhibit very wide host-plant ranges that cover gymnosperm and dicot plants. The Rhizobium radiobacter species complex harbors causative agents of the two diseases. Recently, endophyte isolates from many plant species have been assigned to this species complex. We isolated seven endophyte strains belonging to the species complex from wheat plants and revealed their genomovar affiliations and plasmid profile. The significance of this study is the finding of the genomovar correlation between the endophytes and the known pathogens, the presence of a virulence ability in two of the seven endophyte strains, and the high ratio of the pathogenic strains in the endophyte strains. This study therefore provides convincing evidence that could unravel the mechanism that maintains pathogenic agents of this species and sporadically delivers them to susceptible plants.

Genetic Variation of Multiple Serotypes of Enteroviruses Associated with Hand, Foot and Mouth Disease in Southern China

Enteroviruses (EVs) species A are a major public health issue in the Asia–Pacific region and cause frequent epidemics of hand, foot and mouth disease (HFMD) in China. Mild infections are common in children; however, HFMD can also cause severe illness that affects the central nervous system. To molecularly characterize EVs, a prospective HFMD virological surveillance program was performed in China between 2013 and 2016. Throat swabs, rectal swabs and stool samples were collected from suspected HFMD patients at participating hospitals. EVs were detected using generic real-time and nested reverse transcription-polymerase chain reactions (RT-PCRs). Then, the complete VP1 regions of enterovirus A71 (EV-A71), coxsackievirus A16 (CVA16) and CVA6 were sequenced to analyze amino acid changes and construct a viral molecular phylogeny. Of the 2836 enrolled HFMD patients, 2,517 (89%) were EV positive. The most frequently detected EVs were CVA16 (32.5%, 819), CVA6 (31.2%, 785), and EV-A71 (20.4%, 514). The subgenogroups CVA16_B1b, CVA6_D3a and EV-A71_C4a were predominant in China and recombination was not observed in the VP1 region. Sequence analysis revealed amino acid variations at the 30, 29 and 44 positions in the VP1 region of EV-A71, CVA16 and CVA6 (compared to the respective prototype strains BrCr, G10 and Gdula), respectively. Furthermore, in 21 of 24 (87.5%) identified EV-A71 samples, a known amino acid substitution (D31N) that may enhance neurovirulence was detected. Our study provides insights about the genetic characteristics of common HFMD-associated EVs. However, the emergence and virulence of the described mutations require further investigation.Electronic supplementary materialThe online version of this article (10.1007/s12250-020-00266-7) contains supplementary material, which is available to authorized users.

Genetic diversity of Agrobacterium species isolated from nodules of common bean and soybean in Brazil, Mexico, Ecuador and Mozambique, and description of the new species Agrobacterium fabacearum sp. nov.

Agrobacterium strains are associated with soil, plants and animals, and known mainly by their pathogenicity. We studied 14 strains isolated from nodules of healthy soybean and common bean plants in Brazil, Mexico, Ecuador and Mozambique. Sequence analysis of the 16S rRNA gene positioned the strains as Agrobacterium, but with low phylogenetic resolution. Multilocus sequence analysis (MLSA) of three partial housekeeping genes (glnII, gyrB and recA) positioned the strains in four distinct clades, with Agrobacterium pusense, Agrobacterium deltaense, Agrobacterium radiobacter and Agrobacterium sp. genomospecies G1. Analysis by BOX-PCR revealed high intraspecies diversity. Genomic analysis of representative strains of the three clades indicated that they carry the protelomerase telA gene, and MLSA analysis with six complete housekeeping genes (atpD, glnII, gyrB, recA, rpoB and thrC), as well as average nucleotide identity (less than 90 % with closest species) and digital DNA-DNA hybridization (less than 41 % with closest species) revealed that strain CNPSo 675T and Agrobacterium sp. genomospecies G1 compose a new species. Other phenotypic and genotypic characteristics were determined for the new clade. Although not able to re-nodulate the host, we hypothesize that several strains of Agrobacterium are endophytes in legume nodules, where they might contribute to plant growth. Our data support the description of the CNPSo 675T and Agrobacterium sp. genomospecies G1 strains as a new species, for which the name Agrobacterium fabacearum is proposed. The type strain is CNPSo 675T (=UMR 1457T=LMG 31642T) and is also deposited in other culture collections.

Molecular Characterization of Echinococcus granulosus Isolates Found in Cattle, Buffaloes, Sheep and Goats in Afyonkarahisar, Turkey

This study has been carried out to determine the genotypes of Echinococcus granulosus cysts in cattle, buffaloes, sheep and goats raised in Afyonkarahisar region. Cysts were collected from the internal organs of 258 animals, including 65 goats, 71 sheep, 119 cattle and 3 buffaloes infected with hydatid cysts. DNA was isolated from a total of 78 cysts from germinal membranes and protoscoleces extracted from cysts to identify the genotypes of E. granulosus in infected animals. PCR-RFLP was carried out using the Hin6I and StuI restriction enzymes in the ND1 gene and no polymorphism could be determined in all isolates. In the COX1 gene analysis, G1 strain known as domestic sheep strains and 18 different haplotypes were found in all isolates from cattle, buffaloes, sheep and goats. As a result, it was concluded that all isolates of the cattle, buffaloes, sheep and goats grown in Afyonkarahisar region determined in the analyses carried on the COX1 gene were G1 strain.

Prevalence and molecular characterisation of Echinococcus granulosus in disposed of bovine carcasses in Punjab, India.

Cystic echinococcosis is an important parasitic zoonosis across the globe. The disease is caused by Echinococcus granulosus sensu lato. Free access of dogs to the disposed of bovine carcasses is an important conducive factor for the persistence of E. granulosus in the environment, and thus, stray dogs play a critical role in the disease transmission in developing countries including India. There is lack of information on the status of E. granulosus in the disposed of bovine carcasses in India; however, this information would be helpful to plan future strategies and policies to control this parasite. We examined 85 bovine carcasses from the 7 selected carcasses disposal sites in Punjab. During the postmortem, internal organs were examined for the presence of hydatid cysts. Molecular characterization of Echinococcus spp. was performed by PCR followed by sequencing of partial COI gene. We report 21% (18/85) prevalence of hydatidosis in dispose of bovine carcasses. Age was reported as the significant risk factor (Chi square = 22.5, df = 2, p = 0) for hydatidosis in the studied population and this might be due to higher cumulative exposure to the environmental parasitic stage (E. granulosus eggs) with increase in age. Phylogenetic analysis showed that G1 (predominantly) and G3 strains of E. granulosus are circulating in bovine population in Punjab. Further, studies are needed to determine the strains of E. granulosus circulating in the stray dogs. Measures and policies to combat this problem in India should involve one health approach: a collaborative effort of medical and veterinary physicians.

Molecular epidemiology of rotavirus A and adenovirus among children with acute diarrhea in Hangzhou, China

BackgroundRotavirus A (RVA) and adenovirus (Adv) are important causes of acute diarrhea in children. RVAs are classified into G and P genotypes based on viral proteins (VP)7 and VP4 gene and Adv contains over 70 genotypes based on hexon and fiber gene. This study aimed to characterize the molecular epidemiology of RVA and Adv in children with acute diarrhea during 2017–2018 in Hangzhou.MethodsThe stool samples were collected and tested for RVA and Adv by reverse transcription-quantitative PCR (RT-qPCR) assay. The RVA positive samples were detected by RT-PCR for VP7(G) and VP4([P]) genotypes, and the Adv positive samples were detected by PCR for genotyping by the target to hexon gene.ResultsAmong 228 RVA-positive samples, G9 was detected as the most frequent genotype (195/228, 85.5%), followed by G3 (20/228, 8.8%), G2 (7/228, 3.1%) and G1 (6/228, 2.6%). G9 strains were closely related to strains from China and neighboring countries, as well as the USA. On the other hand, P[8] strains were detected in 219 (96.1%) samples with most closely related to one strain from Malawi, and P[4] in 9 (3.9%) samples. G9P[8] (84.6%, 193/228) was the most prevalent rotavirus A strains, followed by G3P[8] (8.8%, 20/228), G2P[4] (3.1%, 7/228), G1P[8] (2.6%, 6/228) and G9P[4] (0.9%, 2/228). Of 167 Adv-positive cases, 2 different genotypes were identified with 152 (91.0%) of Adv-41and 15 (9%) of Adv-40. All Adv strains were closely related to prototype strains of Adv types 40 and 41 in India.ConclusionsG9P[8] of RVA and Adv-41 were the most common genotypes that caused children’s acute diarrhea in Hangzhou, 2017–2018.

Autophagic Proteome in Two Saccharomyces cerevisiae Strains During Second Fermentation for Sparkling Wine Elaboration.

A correlation between autophagy and autolysis has been proposed in order to accelerate the acquisition of wine organoleptic properties during sparkling wine elaboration. In this context, a proteomic analysis was carried out in two industrial Saccharomyces cerevisiae strains (P29, conventional sparkling wine strain and G1, implicated in sherry wine elaboration) with the aim of studying the autophagy-related proteome and comparing the effect of CO2 overpressure during sparkling wine elaboration. In general, a detrimental effect of pressure and second fermentation development on autophagy-related proteome was observed in both strains, although it was more pronounced in flor yeast strain G1. Proteins mainly involved in autophagy regulation and autophagosome formation in flor yeast G1, and those required for vesicle nucleation and expansion in P29 strain, highlighted in sealed bottle. Proteins Sec2 and Sec18 were detected 3-fold under pressure conditions in P29 and G1 strains, respectively. Moreover, ‘fingerprinting’ obtained from multivariate data analysis established differences in autophagy-related proteome between strains and conditions. Further research is needed to achieve more solid conclusions and design strategies to promote autophagy for an accelerated autolysis, thus reducing cost and time production, as well as acquisition of good organoleptic properties.

Characteristic of panicle in M4 red rice mutants

The need for rice is always increasing every year in line with population growth making rice commodities have a very important role. Red rice which has been known for a long time, needs to be given priority in research to overcome the problem of food and nutrition shortages. Rice panicle, a crucial organ that has economic values, is an important yield component to improve. This study aimed to obtain information on relationship within the panicle characters on rice production. The experiment was conducted within 6 month using 10 red rice mutant lines M4 and 2 non-mutant lines as controls. Data from yield components used were number of grains per panicle, panicle density, percentage of filled grain per panicle, grain length, grain width, and grain thick. Based on the results of the path analysis on these characters, it is known that four characters that have a total positive and significant direct effect on the weight of grain per panicle were the number of grains per panicle, panicle density, panicle length and percentage of filled grain per panicle. Hence, these characters are important and effective to be used as a selection component in increasing grain production in red rice plants. Tests of several M4 generation red rice mutants based on grain characterization show very significant differences in grain length and grain width characters. The longest grain is GK2 strain (10.33 mm) and the widest grain is G9 strain (3.29 mm).

Different Chronic Allograft Pathology Lesions in Two Orthotopic Lung Transplant Rat Strain Combinations

Chronic lung allograft dysfunction (CLAD) remains the major obstacle to long-term survival after lung transplantation. Two different phenotypes have recently been described. About 80% of patients have primarily obstructive lung disease associated with obliterative bronchiolitis, airway fibrosis with intraluminal narrowing (OB-CLAD) while a smaller subset has lung restriction with hypoxemia and parenchymal fibrosis (R-CLAD). Preclinical modeling is essential to better analyze the pathophysiological processes leading to CLAD development. Lung allografts from a large cohort of two different rat strain combinations orthotopically transplanted in the last decade were re-evaluated. From 2007 to 2017 127 rats were transplanted in our centre: 55 Lewis (Lew; RT-1l) into Fisher 344 (F344; RT-1v) (Group 1, G1) and 72 Brown-Norway (BN; RT1n) into Lew (Group 2, G2). Chronic rejection was detected in 39 cases, 18 G1 and 21 G2 rats, respectively, at a mean time of 61 and 17 days, respectively. Several pathological lesions (airway, pleural and vascular fibrosis, airway obliteration, endothelialitis, lymphoid aggregates, epithelial hyperplasia and flattening) were graded following a recent proposed grading system for animal model CLAD (Martinu et al, 2019, AJT). An end stage lung remodelling was detected in 33% of G1 and 48% of G2 models. A vanishing obliterative bronchiolitis (end stage OB-CLAD) was observed in 33% and 20% of G1 and G2 rats, respectively, while signs of fibrosis/fibroelastosis (end stage R-CLAD) were detected in 67% and 80% of G1 and G2 rats, respectively. Acute fibrinous and organizing pneumonia (AFOP) was present in 37.5% of G2 strains and never observed in G1 animals. Interestingly, G2 rats showed higher endothelialitis, vascular fibrosis and lower lymphoid aggregates than G1 strain (p=0.005, p=0.01, p=0.03, respectively). A combination score of highly suggestive R-CLAD lesions (parenchymal, pleural, vascular fibrosis and endothelialitis) was more frequently detected in G2 rats (p=0.03). CLAD associated lesions in lung rat allografts mimic those observed in humans. The earlier R-CLAD developed in G2 may be related to a higher degree of mismatch in this strain combination. These animal models could allow future mechanistic studies to better understand CLAD pathogenesis.

Nine-Year Nationwide Environmental Surveillance of Hepatitis E Virus in Urban Wastewaters in Italy (2011-2019).

Hepatitis E virus (HEV) is an emerging causative agent of acute hepatitis worldwide. To provide insights into the epidemiology of HEV in Italy, a large-scale investigation was conducted into urban sewage over nine years (2011–2019), collecting 1374 sewage samples from 48 wastewater treatment plants located in all the 20 regions of Italy. Broadly reactive primers targeting the ORF1 and ORF2 regions were used for the detection and typing of HEV, followed by Sanger and next generation sequencing (NGS). Real-time RT-qPCR was also used to attempt quantification of positive samples. HEV RNA detection occurred in 74 urban sewage samples (5.4%), with a statistically significant higher frequency (7.1%) in central Italy. Fifty-six samples were characterized as G3 strains and 18 as G1. While the detection of G3 strains occurred in all the surveillance period, G1 strains were mainly detected in 2011–2012, and never in 2017–2019. Typing was achieved in 2 samples (3f subtype). Viral concentrations in quantifiable samples ranged from 1.2 × 103 g.c./L to 2.8 × 104 g.c./L. Our results suggest the considerable circulation of the virus in the Italian population, despite a relatively small number of notified cases, a higher occurrence in central Italy, and a noteworthy predominance of G3 strains.