Ljungan/Sebokele-like picornavirus in birds of prey,common kestrel (Falco tinnunculus) and red-footed falcon (F. vespertinus)

Ljungan and Sebokele viruses are thought to be rodent-borne (picorna)viruses in the genus Parechovirus. Using random amplification and next generation sequencing method a novel Ljungan/Sebokele-like picornavirus was identified in birds of prey. Viral RNA was detected in total of 1 (9%) of the 11 and 2 (28.6%) of the 7 faecal samples from common kestrels and red-footed falcons in Hungary, respectively. High faecal viral RNA load (4.77 × 10⁶ genomic copies/ml) measured by qPCR. The complete genome of picornavirus strain falcon/HA18_080/2014/HUN (KY645497) is 7964-nucleotide (nt) long including a 867-nt 5'end and a 101-nt 3'end (excluding the poly(A)-tail). Falcon/HA18_080/2014/HUN has type-II IRES related to hunnivirus IRES, encodes a polyprotein lacking a leader protein, a VP0 maturation cleavage site and it predicted to encode three 2A proteins (2A₁^NPG↓P, 2A₂^NPG↓P and 2A₃^H-Box/NC), two of them end with ‘ribosome-skipping’ sites (DxExNPG^↓P). Sequence analyses indicated that the ORF1 (6996 nt) polyprotein (2331 amino acid - aa) of falcon/HA18_080/2014/HUN shares the highest aa identity, 59% and 57%, to the corresponding polyproteins of Ljungan and Sebokele viruses. This study reports the identification and complete genome characterization of a novel Ljungan/Sebokele-like picornavirus in faeces of birds of prey which suggests that the genetic diversity and the potential host species spectrum of Ljungan/Sebokele-like viruses in genus Parechovirus are wider than previously thought.     

Detection of a mammalian-like astrovirus in bird,European roller (Coracias garrulus)

Astroviruses are small, non-enveloped viruses with positive sense, single-stranded RNA genomes. The family Astroviridae contains two genera, Mamastrovirus and Avastrovirus, which – based upon our current knowledge – infect mammals and birds, respectively. However, recent seroprevalence study indicated that people with contact to turkeys can develop serological responses to the turkey astrovirus and minks might have been infected with the avastrovirus. These data suggest that the “host species/astrovirus genus” association should be permeable; however, mamastrovirus infection has not been reported from avian species, yet. In this study, a novel astrovirus was identified by viral metagenomics and RT-PCR methods in 2 (11%) out of 19 faecal samples collected from a wild, carnivorous bird species, European rollers (Coracias garrulus) from two breeding territories in Hungary. The complete genome sequence of astrovirus Er/SZAL6/HUN/2011 (KP663426) was 7025 nt-long and had some unique genomic features including an unusually long spacer between the subgenomic RNA promoter and the ORF2 initiation codon. Using the BLASTp Er/SZAL6/HUN/2011 had the highest aa identities 35%, 61% and 34% to MAstV 32 (JF713710, host: porcine), to MAstV 23 (JF729316, host: rabbit) and to unclassified porcine astrovirus (JX684071) in ORF1a, ORF1b and ORF2, respectively. The same proteins of Er/SZAL6/HUN/2011 had 25%, 66% and 33% aa identities to the corresponding proteins of murine astrovirus (JX544743) as the closest strain. The sequence- and phylogenetic analysis indicated that Er/SZAL6/HUN/2011 represents the first member of a novel mamastrovirus species. Data suggest that both mammals and birds could have been exposed to mamastroviruses and avastroviruses providing opportunities for cross-species infection and viral adaptation with cross-class astroviruses especially in carnivorous animals. Further investigation is needed to determine the origin, natural host species spectrum, distribution and spread of Er/SZAL6/HUN/2011 among vertebrates.     

Antigenic composition and immunoreactivity differences between HEV recombinant capsid proteins generated from different genotypes

Appreciable variability has been observed in hepatitis E virus (HEV) serological diagnostics. Four recombinant proteins (p166s) were generated from position 452 to 617 aa of ORF2 of different HEV genotypes and used in an indirect ELISA to detect anti-HEV IgMs and IgGs in serially diluted sera of patients infected with different HEV genotypes (genotype 1, n = 15; genotype 3, n = 12; genotype 4, n = 17). To evaluate the differences at a conformational level, 3D-structure models of p166s were predicted, and different bioinformatics tools were used to analyze the antigenic composition. With both anti-HEV IgMs and IgGs antibodies, there was a considerable variability between the four antigens immunoreactivities. In silico results revealed the region 483–533 aa with the highest antigenic potential and contains six key aa at positions 488, 489, 512, 533, 483 and 530. This immunoreactivity variation could affect diagnosis results and seroprevalence estimations and the identification in silico of a region highly antigenic would guide the development of efficient serological assays and epitope-based vaccines.     

Evaluation of recombinant Chinese avian hepatitis E virus (CaHEV) ORF2 and ORF3 proteins for protection of chickens against CaHEV infection

Avian hepatitis E virus (HEV) is the etiologic agent of big liver and spleen disease in chickens. In 2010, the Chinese avian HEV (CaHEV) strain was isolated from chickens and demonstrated to cause the decreased egg production in layer hens. No avian HEV commercial vaccine has yet been developed to prevent virus infection in China. In this study, recombinant CaHEV truncated ORF2 and complete ORF3 proteins were evaluated separately for immunoprotection of chickens against CaHEV infection. First, truncated ORF2 and complete ORF3 proteins were expressed in Escherichia coli. Next, 48 specific-pathogen-free chickens were randomly divided into three groups. One group was immunized with truncated ORF2 protein, the second group was immunized with recombinant ORF3 protein, while the third group (control) was mock-immunized with PBS. After booster immunization, chickens in all three groups were challenged intravenously with CaHEV infectious stock and assessed for viremia, fecal virus shedding, seroconversion, and gross hepatic lesions. In the ORF2 protein-immunized group, no chickens showed evidence of avian HEV infection. In the ORF3 protein-immunized group, nine chickens exhibited viremia and seven had fecal virus shedding. In the control group, all 16 chickens showed viremia and fecal virus shedding. However, the durations in chickens from the ORF3 protein group (2–4 weeks) were shorter than the ones from the control group (4–8 weeks). Moreover, no gross liver lesions emerged in the ORF2 protein group, while lesions observed in the ORF3 protein group were milder than in controls. Therefore, the ORF2 protein can confer complete immunoprotection against chicken CaHEV infection, while the ORF3 protein only confers partial immunoprotection.     

Genetically highly divergent RNA virus with astrovirus-like (5′-end) and hepevirus-like (3′-end) genome organization in carnivorous birds,European roller (Coracias garrulus)

Astroviruses (family Astroviridae) and hepeviruses (family Hepeviridae) are small, non-enveloped viruses with genetically diverse +ssRNA genome thought to be enteric pathogens infecting vertebrates including humans. Recently, many novel astro- and hepatitis E virus-like +ssRNA viruses have been described from lower vertebrate species. The non-structural proteins of astro- and hepeviruses are highly diverse, but the structural/capsid proteins represent a common phylogenetic position shed the light of their common origin by inter-viral recombination. In this study, a novel astrovirus/hepevirus-like virus with +ssRNA genome (Er/SZAL5/HUN/2011, MK450332) was serendipitously identified and characterized from 3 (8.5%) out of 35 European roller (Coracias garrulus) faecal samples by RT-PCR in Hungary. The complete genome of Er/SZAL5/HUN/2011 (MK450332) is 8402 nt-long and potentially composed three non-overlapping open reading frames (ORFs): ORF1a (4449 nt/1482aa), ORF1b (1206 nt/401aa) and ORF2 (1491 nt/496aa). The ORF1ab has an astrovirus-like genome organization containing the non-structural conserved elements (TM, CC, NLS, VPg) and enzyme residues (trypsine-like protease, RNA-dependent RNA-polymerase) with low amino acid sequence identity, 15% (ORF1a) and 44% (ORF1b), to astroviruses. Supposedly the ORF2 is a capsid protein but neither the astrovirus-like subgenomic RNA promoter (sgRNA) nor the astrovirus-like capsid characteristics have been identifiable. However, the predicted capsid protein (ORF2) showed 26% identity to the corresponding protein of hepevirus-like novel Rana hepevirus (MH330682). This novel +ssRNA virus strain Er/SZAL5/HUN/2011 with astrovirus-like genome organization in the non-structural genome regions (ORF1a and ORF1b) and Rana hepevirus-related capsid (ORF2) protein represent a potentially recombinant virus species and supports the common origin hypothesis, although, the taxonomic position of the studied virus is still under discussion.     

Évolution du statut nutritionnel d’une cohorte d’enfants tunisiens au cours de l’hospitalisation : facteurs de risque de la dénutrition hospitalière

ObjectivesStudy the evolution of the nutritional status of a cohort of hospitalized children and identify the risk factors of hospital malnutrition.MethodsProspective, cross-sectional study carried out in a pediatric department over a period of six months, including all children aged ≥ 30 days, hospitalized for a period ≥ six days. Anthropometric data were assessed on admission and discharge. Food consumption was assessed using the flower tool. We identified the risk factors for hospital undernutrition (HUN) by multivariate analysis.ResultsWe included 120 patients with a mean age 46.3 months. The prevalence of acute undernutrition at admission was 21.7% and that of chronic undernutrition was 10%. The prevalence of acute undernutrition at discharge rose to 34%. Weight loss during hospitalization was noted in 68.3% of cases. The prevalence of HUN was 55% considering a decrease in BMI or P/PAT z-score ≥ 25%. The risk factors for HUN were: age ≤ 24 months (P = 0.039; OR 95% CI = 2.67 [1.05–6.82]), the presence of undernutrition on admission (P = 0.002; OR 95% CI = 2.32 [0.93–6.51]) and average food consumption < 50% during hospitalization (P < 10^?3; OR 95% CI = 6.69 [2.57–17.40]).ConclusionScreening for undernutrition on admission to hospital as well as assessment of the nutritional risk in hospitalized children is essential so that preventive or curative nutritional care can be taken.     

Isolation and genomic characterization of a classical Muscovy duck reovirus isolated in Zhejiang,China

A classical Muscovy reovirus was isolated from a sick Muscovy duck with white necrotic foci in its liver in Zhejiang, China, in 2000. This classical reovirus was propagated in a chicken fibroblast cell line (DF-1) with obvious cytopathic effects. Its genome was 22,967 bp in length, with approximately 51.41% G + C content and 10 dsRNA segments encoding 11 proteins, which formed a 3/3/4 electrophoretic PAGE profile pattern. The length of the genomic segments was similar to those of avian orthoreoviruses (ARV and N-MDRV), ranging from 3959 nt (L1) to 1191 nt (S4). All of the segments have the conserved terminal sequences 5′-GCUUUU——UUCAUC-3′, and with the exception of the S4 segment, all the genome segments apparently encode one single primary translation product. The genome analysis revealed that the S4 segment of classical MDRV is a bicistronic gene, encoding the overlapping ORFs for p10 and σC but distinct from ARV and N-MDRV/N-GRV, which codes for p10, p18 and σC via the tricistronic S1 segment. A comparative sequence analysis provided evidence indicating extensive sequence divergence between classical MDRV and other avian orthoreoviruses. A phylogenetic analysis based on the RNA-dependent RNA polymerase (RdRp) and the major outer capsid proteins σC was performed. Members of the DRVs in the Avian orthoreovirus species were clustered into two genetic groups (classical MDRV and N-MDRV genotype), and the classical MDRV isolates formed distinct lineages (China and Europe lineages), suggesting that the classical MDRVs isolated in restricted geographical region are evolving by different and independent pathways.     

Prevalence and complete genome characterization of turkey picobirnaviruses

The “light turkey syndrome” (LTS), in which birds weigh less than their standard breed character at the marketing time, is believed to be a consequence of viral enteritis at an early age (3–5 weeks) from which the birds never fully recover. In a previously published study, we collected fecal pools from 2, 3, 5 and 8 week old turkey poults (80 pools from LTS farms and 40 from non-LTS farms) and examined them for the presence of astro-, rota-, reo-, and coronaviruses. To determine the presence of additional enteric viruses, we analyzed a fecal pool by Illumina sequencing and found picobirnavirus (PBV). Segments 1 and 2 of this virus shared 45.8% aa and 60.9–64.5% aa identity with genogroup I of human PBV, respectively. Primers based on RNA-dependent RNA polymerase and capsid genes were designed for detection and molecular characterization of PBVs in the 120 fecal pools described above. From LTS farms, 39 of 80 (48.8%) pools were PBV positive while 23 of 40 (57.5%) were positive from non-LTS farms. The phylogenetic analysis of 15 randomly selected strains divided them into four subgroups within genogroup I (subgroups 1A–D). Nine strains were in subgroup IA showing 69.9–76.4% nt identity with human PBV GI strainVS111 from the Netherlands. Strains in subgroup IB (n = 2) had 91.4–91.7% nt identity with chicken PBV GI strain AVE 42v1 from Brazil. Two strains in subgroup IC had 72.3–74.2% nt identity with chicken PBV strain AVE 71v3 from Brazil. In subgroup ID, two strains showed 72.4–81.8% nt identity with chicken PBV GI strain AVE 57v2 from Brazil. Subgroup IC and ID were the most divergent. Five of the 15 strains were typed using capsid gene primers. They showed 32.6–33.4% nt and 39.5–41.3% aa identity with VS10 human PBV strain. These results indicate co-circulation of divergent strains of PBVs among Minnesota turkeys.     

Reverse spillover of avian viral vaccine strains from domesticated poultry to wild birds

Transmission of viruses from the commercial poultry to wild birds is an emerging paradigm of livestock–wildlife interface. Here, we report the identification and isolation of vaccine strains of avian paramyxovirus serotype 1 (APMV1) and avian coronaviruses (ACoV) from different wild bird species across eight Egyptian governorates between January 2014 and December 2015. Surveillance of avian respiratory viruses in free-ranging wild birds (n = 297) identified three species that harboured or excreted APMV1 and ACoVs. Genetic characterization and phylogenetic analysis of recovered viruses revealed a close association with the most widely utilized vaccine strains in the country. These results highlight the potential spillover of vaccine-viruses probably due to extensive use of live-attenuated vaccines in the commercial poultry, and close interaction between domesticated and wild bird populations. Further exploring the full spectrum of vaccine-derived viral vaccine strains in wild birds might help to assess the emergence of future wild-birds origin viruses.     

Comparison of hepatitis E virus genotypes from rabbits and pigs in the same geographic area: No evidence of natural cross-species transmission between the two animals

Domesticated pigs have been shown to be a reservoir of genotypes 3 and 4 hepatitis E virus (HEV). Farmed rabbits were recently recognized as the host of a novel virus, rabbit HEV. In order to determine whether HEV is transmitted naturally between rabbits and pigs, a survey on HEV infections was conducted in rabbits and pigs aged 2–4 months from rabbit and pig farms located near to each other in nine villages in three counties of Hebei Province, China. The overall anti-HEV antibody positivity rates in serum samples of swine and rabbits were 61.7% (58/94) and 23.2% (67/289), and the positive rates for HEV RNA were 23.4% (22/94) and 10% (29/289), respectively. In addition, 37 of 125 swine fecal samples (29.6%) were HEV RNA positive. The nucleotide sequences of a 304 bp region within HEV ORF2 have identity ranging from 84.5% to 100% among the rabbit isolates and from 82.3% to 100% among the swine isolates. In contrast, the nucleotide identity between the two species groups was only 72–76.6%. Consequently, the two groups were clearly separated in the phylogenetic tree that showed all of the rabbit isolates are closely related to the rabbit HEV reported recently and the swine isolates belong to genotype 4, including subgenotypes 4a, 4c and 4d. The results showed that HEV is highly prevalent in farmed rabbits and pigs in these areas. However, genotype 4 HEV and rabbit HEV are circulating separately in pigs and rabbits in the same area. In conclusion, there was no evidence of cross-species transmission of HEV between pigs and rabbits. The frequency of HEV transmission events between these two animal species is likely low in commercial farms.     

Further circulation of West Nile and Usutu viruses in wild birds in Italy

Usutu virus (USUV) and West Nile virus (WNV) are emerging pathogens that can cause neurological disease in humans. From March 2012 to June 2013, a sero-survey on wild birds was carried out to investigate the circulation of both viruses in Northwest (NW) Italy. Samples belonging to 47 different bird species have been collected using a volunteer based network and a wildlife rehabilitation center. Four of 297 serum samples had neutralizing antibodies against USUV (P = 1.34%, IC 95% 0.36–3.4), while 10 of 233 samples tested positive for WNV (P = 4.29%, IC 95% 2.07–7.75). Neutralizing antibodies for WNV were significantly more prevalent (p < 0.001) in trans-Saharan migrants (P = 21%, IC 95% 9.55–37.3) than in resident and short-distance birds, but no migratory habit-related differences were found for USUV. Antibodies in resident bird species suggest that both viruses are circulating in NW Italy.     

Emerging trends in the epidemiology of human astrovirus infection among infants,children and adults hospitalized with acute watery diarrhea in Kolkata,India

Human astroviruses (HAstVs) have now emerged as another common cause of non-bacterial acute gastroenteritis (AGE) in humans worldwide. This study investigated the epidemiology and genetic diversity of human astrovirus strains circulating among infants, younger children (up to 6 years), older children and adolescents (>6–17 years) and adults (18 years and above) hospitalized for diarrhea and their role in AGE in Kolkata, India. A total of 2535 fecal samples were screened for the presence of known enteric viral, bacterial and parasitic etiologies by conventional microbiological assays and molecular methods. The overall incidences of sole or mixed infection of HAstV with known enteric viral, bacterial and parasitic pathogens were detected in 60 cases (2.4%) among all age groups. The clinical symptoms of astrovirus-associated acute watery diarrhea cases were recorded for all sole and mixed infection cases. A high number of sole (n = 13/60 [21.7%]) and mixed infection cases (n = 22/60 [36.7%]) were observed in adults (18 years old or more). Considering all age groups, 18 sole infection cases (n = 18/60 [30%]) and 42 mixed infection cases (n = 42/60 [70%]) with Rotavirus (n = 11/25 [44%]), Vibrio cholerae O1 (n = 6/24 [25%]) Cryptosporidium spp and Giardia lamblia (n = 5/13 [38.4%]) were observed. Further, eleven HAstV samples from infants and children (up to 6 years), children and adolescents (>6–17 years) and adults (18 years and above) were analyzed for their sequences of overlap region between ORF1b (RdRp) and ORF2 (capsid). Among these, ten strains were found to have close genetic relatedness to the Japanese strain HAstV_G1 [AB009985]. Additionally, the IDH2211 Kolkata strain showed a close genetic match with the Thai HAstV_G3 strain [EU363889]. Our study reports show that HAstVs as the sole agent and as mixed infection with other known enteric viral, bacterial, parasitic pathogens are also responsible for AGE among infants, children, adolescents and adults in Kolkata, India.     

Environmental implication of iodine in water,milk and other foods used in Saharawi refugees camps in Tindouf,Algeria

A cross-sectional survey among Saharawi refugees in four camps carried out in 2007 revealed enlarged thyroid volume and high urinary iodine concentration in women and school children. The purpose of this paper is to describe the content of iodine in food and water and explore whether any sources in the environment can explain the situation. Samples of water (n = 143), milk (n = 19) and salt (n = 89) were collected. Different wells supplied the camps with water and the median iodine concentration was 108 μg/L (range 55–545 μg/L) and significantly higher in two of the camps (El Aiune and Ausserd; 300 μg/L (range 55–545 μg/L)), compared to the two other camps (Smara; 87 μg/L (55–127 μg/L) and Dakla; 70 μg/L (55–96 μg/L)). In local goat milk the median iodine concentration was 370 μg/L (70–13,070 μg/L). The median content of iodine in salt was 6 μg/g (0–51 μg/g). Water and local milk were the most important sources of iodine for women. High levels of iodine in water seem to be one of the main sources of iodine that affects humans as well as animals.     

Adjuvant potential of resiquimod with inactivated Newcastle disease vaccine and its mechanism of action in chicken

Resiquimod (R-848), an imidazoquinoline compound, is a potent synthetic Toll-like receptor (TLR) 7 agonist. Although the solitary adjuvant potential of R-848 is well established in mammals, such reports are not available in avian species hitherto. Hence, the adjuvant potential of R-848 was tested in SPF chicken in this study. Two week old chicks were divided into four groups (10 birds/group) viz., control (A), inactivated Newcastle disease virus (NDV) vaccine prepared from velogenic strain (B), commercial oil adjuvanted inactivated NDV vaccine prepared from lentogenic strain (C) and inactivated NDV vaccine prepared from velogenic strain with R-848 (D). Booster was given two weeks post primary vaccination. Humoral immune response was assessed by haemagglutination inhibition (HI) test and ELISA while the cellular immune response was quantified by lymphocyte transformation test (LTT) and flow cytometry post-vaccination. Entire experiment was repeated twice to check the reproducibility. Highest HI titre was observed in group D at post booster weeks 1 and 2 that corresponds to mean log₂ HI titre of 6.4 ± 0.16 and 6.8 ± 0.13, respectively. The response was significantly higher than that of group B or C (P < 0.01). LTT stimulation index (P ≤ 0.01) as well as CD4⁺ and CD8⁺ cells in flow cytometry (P < 0.05) were significantly high and maximum in group D. Group D conferred complete protection against virulent NDV challenge, while it was only 80% in group B and C. To understand the effects of R-848, the kinetics of immune response genes in spleen were analyzed using quantitative real-time PCR after R-848 administration (50 μg/bird, i.m. route). Resiquimod significantly up-regulated the expression of IFN-α, IFN-β, IFN-γ, IL-1β, IL-4, iNOS and MHC-II genes (P < 0.01). In conclusion, the study demonstrated the adjuvant potential of R-848 when co-administered with inactivated NDV vaccine in SPF chicken which is likely due to the up-regulation of immune response genes.     

Report of a novel C1483W mutation in the hepatitis E virus polymerase in patients with acute liver failure

Here, we report the molecular alterations in the HEV genome from patients with acute liver failure (ALF) and acute viral hepatitis (AVH) from North India, including pregnant women and its association with the poor outcome of the disease. We partially sequenced the RNA Dependent RNA polymerase (RdRp) region of the ORF 1 protein in the HEV genome from representative samples from patients with ALF and AVH and identified two novel mutations Cysteine 1483 Tryptophan and Asparagine 1530 Threonine in 100% (25/25) of the patients with ALF compared to none (0/30) of the patients with AVH (P < 0.0001). Disease severity parameters along with viral load corresponding to the samples with C1483W and N1530T mutations were significantly higher compared to those lacking the mutation showing significant association with the outcome in ALF patients. The nucleotide substitutions in the RdRp region may play a crucial role in enhancing HEV replication thus leading to disease severity.     

Prospective effects of pedometer use and class competitions on physical activity in youth: A cluster-randomized controlled trial

ObjectiveTo evaluate the immediate effects of a school-based multi-component program to foster a physically active lifestyle in adolescence.Design/participantsIn a cluster-randomized controlled trial with pre- and post-assessment in 2014, 29 schools with 1162 8th grade students (48% girls) from Germany were included. Age ranged from 12 to 17 years (M = 13.74; SD = 0.67).InterventionWhile the control group attended education as usual, students in the intervention group received pedometers and took part in a class competition over a time period of 12 weeks. Classes with the most steps and best creative ideas to promote physical activity in everyday life were awarded.Main outcome measuresPrimary outcomes included out-of-school sports activities (h/week), moderate to vigorous physical activity (days/week with a minimum of 60 min), active commuting (min/day), doing chores (min/day), and sedentary behavior (h/day) assessed through self-administered questionnaires as well as cardiorespiratory fitness measured using the 20-m shuttle-run test (completed laps).ResultsSignificant interaction terms between group and wave of assessment were found on out-of-school sports activities (b = − 1.09 [− 1.89; − 0.29], p = 0.008), moderate to vigorous physical activity (b = − 0.29 [− 0.47; − 0.10], p = 0.002), and active commuting (b = − 20.41 [− 32.32; − 8.49], p = 0.001): students in the intervention group showed a higher increase of physical activity levels than students in the control group. The intervention effect on cardiorespiratory fitness missed significance marginally (b = − 1.52 [− 3.14; 0.98], p = 0.065), There was no effect on students' sedentary behavior (b = 0.06 [− 0.72; 0.84], p = 0.881).ConclusionsAn easy to administer school-based physical activity program (12 weeks) may enhance students' leisure-time physical activity.Trial registration number: ISRCTN49482118     

Toll-like receptor 8 (TLR8) polymorphisms are associated with non-progression of chronic hepatitis C in HIV/HCV coinfected patients

Toll-like receptor 8 (TLR8) polymorphisms have been related to hepatitis C virus (HCV) infection. The aim was to estimate the association of TLR8 polymorphisms with HCV-related outcomes in HIV/HCV coinfected patients. We performed a cross-sectional study of 220 patients who underwent a liver biopsy. TLR8 polymorphisms were genotyped using GoldenGate® assay. The outcome variables were non-fibrosis (F0), mild-inflammation (A0/A1), and non-steatosis [fatty hepatocytes (FH) < 10%]. Logistic regression analysis was used to compare the outcome variables according to TLR8 polymorphisms. Four polymorphisms were analyzed (rs1013151, rs5744069, rs17256081 and rs3764880rs1013151). Female patients had higher frequency of TLR8 major alleles at rs17256081 and rs101315, and minor alleles at rs3764880 and rs5744069. Male patients had higher frequency of TLR8 minor alleles except for rs3764880, where major alleles were higher (p < 0.01). Two TLR8 polymorphisms (rs1013151 and rs5744069) were significantly associated with non-fibrosis (F0) [adjusted odds ratio (aOR) = 4.42 (95% of confidence interval (95%CI) = 1.54; 12.68) (p = 0.006) and aOR = 4.76 (95%CI = 1.69; 13.37) (p = 0.003); respectively]. When data were stratified by gender, rs1013151 and rs5744069 polymorphisms remained significant for male patients [adjusted odds ratio (aOR) = 4.49 (95%CI = 1.08; 18.62) (p = 0.039) and aOR = 6.17 (95%CI = 1.45; 26.20) (p = 0.014); respectively]. When data were stratified by major HCV genotypes, patients infected with HCV genotype 1 (GT1) had significant values for both rs1013151 and rs5744069 polymorphisms [aOR = 5.79 (95%CI = 1.44; 23.32) (p = 0.013) and aOR = 8.01 (95%CI = 2.16; 35.65) (p = 0.005); respectively]. Finally, none of the TLR8 polymorphisms were significantly associated with mild-inflammation or non-steatosis. In conclusion, TLR8 polymorphisms seem to be related to non-progression of liver fibrosis in HIV/HCV coinfected patients, particularly in males and those patients infected with GT1.     

A novel feline norovirus in diarrheic cats

By screening a collection of fecal samples from young cats housed in three different shelters in South Italy, noroviruses (NoVs) were found in 3/48 (6.2%) specimens of animals with enteritis signs while they were not detected in samples collected from healthy cats (0/57). Upon sequence analysis of the short RNA-dependent RNA polymerase (RdRp) region, the three strains displayed the highest nucleotide (nt) and amino acid (aa) identities to the prototype GIV.2 strain lion/Pistoia/387/06/ITA (91.0–93.0% nt and 97.0–98.0% aa). The sequence of ~ 3.4-kb portion at the 3′ end of the genome of a NoV strain, TE/77-13/ITA, was determined. In the full-length ORF2, encoding the VP1 capsid protein, the virus was genetically closest to the canine GVI.2 NoV strains C33/Viseu/2007/PRT and FD53/2007/ITA (81.0–84.0% nt and 93.0–94.0% aa identities), suggesting a recombination nature, with the cross-over site being mapped to the ORF1-ORF2 junction. Based on the full-length VP1 amino acid sequence, we classified the novel feline NoV, together with the canine strains Viseu and FD53, as a genotype 2, within the genogroup GVI. These findings indicate that, as observed for GIV NoV, GVI strains may infect both the canine and feline host. Unrestricted circulation of NoV strains in small carnivores may provide the basis for quick genetic diversification of these viruses by recombination. Interspecies circulation of NoVs in pets must also be considered when facing outbreaks of enteric diseases in these animals.