Genetic Variability of Autochthonous Pear Varieties from Tuzla Canton, Bosnia and Herzegovina

Pear as a fruit species has a special place because of its quality characteristic. The fact that about 20,000 pear seedlings are considered to be sold annually in Bosnia and Herzegovina, which are autochthonous or spontaneously expanded varieties, also speaks in favor. The main goal of this research is to analyze the genetic variability of nine autochthonous pear varieties in Tuzla Canton, Bosnia and Herzegovina to enable the conservation and expansion of existing genetic resources. The study included nine autochthonous pear varieties. Samples of young leaves were collected on the following localities: the town of Srebrenik, the town of Gradačac, the municipality of Čelić-Tuzla Canton, Bosnia and Herzegovina, in the spring 2019. To determine genetic diversity, 12 SSR (Simple Sequence Repeats) markers were genotyped. There were no cases of synonyms or homonyms in the analyzed set. Each of the nine tested varieties represents a unique genotype. Autochthonous pear cultivars analyzed in this paper represent an interesting genetic resource, with useful agronomic traits that can be used in future cultivation.

A unique genotype of pseudohypoaldosteronism type 1b in a highly consanguineous population

Context This study describes the molecular genetics of pseudohypoaldosteronism type 1b (PHA 1b) in the highly consanguineous population of two Arabian Gulf countries, Saudi Arabia and Oman. Patients and Methods This study enrolled 22 patients from 13 unrelated families (two families with five patients from Oman and 11 families with 17 patients from Saudi Arabia). All of these patients had presented within the first 10 days of life with nausea and vomiting, hyponatremia, hyperkalemia and hypotension. We isolated DNA from peripheral blood and PCR-sequenced all exons and exon-intron boundaries of SCNN1A and, if negative, SCNN1B and SCNN1G using the Dideoxy Chain termination method. Results We found a total of eight mutations in 13 families as follows: six mutations in SCNN1A, one in SCNN1B, and one in SCNN1G. All of these mutations were novel except one. SCNN1A mutations were: c.1496A>G, p.Q499R (novel) in one patient; c.1453C>T, p.Q485X (novel) in one patient; c.1322_1322delA, p.N441Tfs*41 (novel) in two patients of one family; c.876 + 2 delGAGT (novel) in three patients of one family; c.203_204 delTC, p.I68Tfs*76 (known mutation) in eight patients of five families; and whole SCNN1A gene deletion (novel) in two patients of two families. In addition, a nonsense SCNN1B mutation c.1694C>A, p.S565X (novel) was found in three siblings from one Omani family, and an SCNN1G deletion mutation c.527_528 delCA, p.T176Rfs*9 (novel) in two siblings form another Omani family. Conclusion We characterized a unique genotype of PHA 1b with several novel gene-structure disrupting mutations in SCNN1A, SCNN1B and SCNN1G in a consanguineous population.

Novel Clade 2.3.4.4b Highly Pathogenic Avian Influenza A H5N8 and H5N5 Viruses in Denmark, 2020

Since late 2020, outbreaks of H5 highly pathogenic avian influenza (HPAI) viruses belonging to clade 2.3.4.4b have emerged in Europe. To investigate the evolutionary history of these viruses, we performed genetic characterization on the first HPAI viruses found in Denmark during the autumn of 2020. H5N8 viruses from 14 wild birds and poultry, as well as one H5N5 virus from a wild bird, were characterized by whole genome sequencing and phylogenetic analysis. The Danish H5N8 viruses were found to be genetically similar to each other and to contemporary European clade 2.3.4.4b H5N8 viruses, while the Danish H5N5 virus was shown to be a unique genotype from the H5N5 viruses that circulated at the same time in Russia, Germany, and Belgium. Genetic analyses of one of the H5N8 viruses revealed the presence of a substitution (PB2-M64T) that is highly conserved in human seasonal influenza A viruses. Our analyses showed that the late 2020 clade 2.3.4.4b HPAI H5N8 viruses were most likely new incursions introduced by migrating birds to overwintering sites in Europe, rather than the result of continued circulation of H5N8 viruses from previous introductions to Europe in 2016/2017 and early 2020.

First isolation and whole-genome characterization of a G9P[14] rotavirus strain from a diarrheic child in Egypt

An unusual group A rotavirus (RVA) strain (RVA/Human-tc/EGY/AS997/2012/G9[14]) was isolated for the first time in a faecal sample from a 6-month-old child who was hospitalized for treatment of acute gastroenteritis in Egypt in 2012. Whole-genome analysis showed that the strain AS997 had a unique genotype constellation: G9-P[14]-I2-R2-C2-M2-A11-N2-T1-E2-H1. Phylogenetic analysis indicated that the strain AS997 had the consensus P[14] genotype constellation with the G9, T1 and H1 reassortment. This suggests either a mixed gene configuration originated from a human Wa-like strain and a P[14]-containing animal virus, or that this P[14] could have been acquired via reassortment of human strains only. The study shows the possible roles of interspecies transmission and multiple reassortment events leading to the generation of novel rotavirus genotypes and underlines the importance of whole-genome characterization of rotavirus strains in surveillance studies.

Autism-like behaviors in male mice with a Pcdh19 deletion

Mutations in protocadherin 19 (PCDH19), which is on the X-chromosome, cause the brain disease Epilepsy in Females with Mental Retardation (EFMR). EFMR is also often associated with autism-like symptoms. In mice and humans, epilepsy occurs only in heterozygous females who have a mixture of PCDH19 wild-type (WT) and mutant cells caused by random X-inactivation; it does not occur in hemizygous PCDH19 mutant males. This unique inheritance pattern strongly suggests the underlying disease mechanism operates via interference between WT and mutant cells rather than being a result of complete loss of PCDH19 functions. Although it remains unclear whether the other symptoms of EFMR also conform to this unique genotype-phenotype relationship, PCDH19 mutant males were recently reported to demonstrate autism-like symptoms. We, therefore, used a Pcdh19 knockout (KO) mouse model to ask whether a complete lack of PCDH19 causes autism-like behaviors. Consistent with the autism observed in EFMR females, we found Pcdh19 heterozygous KO female mice (with mosaic expression of PCDH19) show defects in sociability in the 3-chamber test. Surprisingly, hemizygous Pcdh19 KO male mice (without any PCDH19 expression) exhibit impaired sociability in the 3-chamber test and reduced social interactions in the reciprocal social interaction test. We also observed that, compared to WT mice, mutant mice display more repetitive behaviors, including self-grooming and rearing. These findings indicate that hemizygous Pcdh19 KO male mice show autism-like phenotypes.

2269. Clinical Outcomes in Patients with Carbapenem-Non-Susceptible, β-Lactam-Susceptible Pseudomonas aeruginosa Infections

Background Pseudomonas aeruginosa (PsAr) isolates harboring OprD mutations often present phenotypically as carbapenem nonsusceptible but susceptible to antipseudomonal β-lactams (APBLs). It is unknown whether this unique genotype–phenotype combination affects the clinical outcomes of patients infected with these pathogens. The objective of this study was to compare clinical outcomes of patients treated with APBLs for pneumonia and/or bacteremia caused by PsAr bearing this unique carbapenem nonsusceptible, β-lactam susceptible phenotype (Carba-NS) to those retaining susceptibility to both carbapenems and APBLs (Carba-S). Methods Retrospective multicenter cohort of adult in-patients who received effective APBL for PsAr pneumonia and/or bacteremia from January 2012 to November 2018. Baseline characteristics, treatment information, and clinical outcomes were obtained from the electronic medical record. The primary outcome was 14-day mortality. Secondary outcomes included 30-day mortality, 30-day infection recurrence, and infection-related length of stay (IR-LOS). IR-LOS was defined as the time from index culture to antibiotic discontinuation or hospital discharge, whichever was sooner. Descriptive statistics were analyzed using SPSS. Results 387 patients were evaluated; 60 Carba-S and 21 Carba-NS were included. The primary reason for exclusion was ineffective empiric therapy. Select demographics and clinical outcomes are displayed in Table 1. Compared with the Carba-S group, Carba-NS patients were younger, had better renal function, increased incidence of pneumonia, more severely ill, and higher rate of empiric ceftazidime use. Despite these differences at baseline there were no significant differences in empiric APBL treatment patterns, 14- or 30-day mortality, or recurrence at 30 days between the groups. Carba-NS patients had lower rate of oral step down therapy and a significantly longer LOS and IR-LOS. Conclusion In this cohort of patients who received appropriate and timely APBL therapy, the Carba-NS phenotype was not associated with increased rates of 14-day mortality, 30-day mortality, or 30-day infection recurrence. These data suggest that APBLs may be effective therapeutic options against this phenotype. Further research is warranted to confirm these findings. Disclosures All authors: No reported disclosures.

Presence of a Novel Subtype of Bovine Hepacivirus in China and Expanded Classification of Bovine Hepacivirus Strains Worldwide into 7 Subtypes

The newest member of the Hepacivirus genus, bovine hepacivirus (BovHepV), was first identified in cattle in 2015 and is a novel hepacivirus C virus (HCV)-like virus. This virus has been detected in five countries so far and is classified into four subtypes. Bovine serum is commonly used for cell cultures and is considered the major source of viral contamination of pharmaceutical products. In this study, bovine serum samples were collected from seven countries located in Asia, America, Oceania, and Europe and were tested for BovHepV RNA using nested PCR, in order to: (i) obtain more knowledge on the geographical distribution and subtypes of BovHepV; and (ii) detect the potential contamination of BovHepV in commercial bovine serum samples used for cell culture propagation. The results demonstrated that bovine serum samples from individual donor cattle in China contained BovHepV RNA. After PCR, sequencing, and assembly, the genomes of the Chinese BovHepV strains were obtained. Genetic analysis of the polyprotein gene revealed a protein identity of <77% and a nucleotide identity of <85% between the Chinese BovHepV strains and all other previously reported BovHepV strains. Using cut-off values for determination of HCV genotypes and subtypes, BovHepV strains worldwide were classified into one unique genotype and seven subtypes. The BovHepV strains identified in the present study were classified into a novel subtype, which was provisionally designated subtype G. The genetic relationships among the different BovHepV subtypes were further confirmed through phylogenetic analysis. The present study provides critical insights into BovHepV’s geographical distribution and genetic variability.

Employing toxin-antitoxin genome markers for identification of Bifidobacterium and Lactobacillus strains in human metagenomes

Recent research has indicated that in addition to the unique genotype each individual may also have a unique microbiota composition. Difference in microbiota composition may emerge from both its species and strain constituents. It is important to know the precise composition especially for the gut microbiota (GM), since it can contribute to the health assessment, personalized treatment, and disease prevention for individuals and groups (cohorts). The existing methods for species and strain composition in microbiota are not always precise and usually not so easy to use. Probiotic bacteria of the genus Bifidobacterium and Lactobacillus make an essential component of human GM. Previously we have shown that in certain Bifidobacterium and Lactobacillus species the RelBE and MazEF superfamily of toxin-antitoxin (TA) systems may be used as functional biomarkers to differentiate these groups of bacteria at the species and strain levels. We have composed a database of TA genes of these superfamily specific for all lactobacilli and bifidobacteria species with complete genome sequence and confirmed that in all Lactobacillus and Bifidobacterium species TA gene composition is species and strain specific. To analyze composition of species and strains of two bacteria genera, Bifidobacterium and Lactobacillus, in human GM we developed TAGMA (toxin antitoxin genes for metagenomes analyses) software based on polymorphism in TA genes. TAGMA was tested on gut metagenomic samples. The results of our analysis have shown that TAGMA can be used to characterize species and strains of Lactobacillus and Bifidobacterium in metagenomes.