SNP Analysis Research Articles

Evaluation of nationwide analysis surveillance for methicillin-resistant Staphylococcus aureus within Genomic Medicine Sweden.

Background. National epidemiological investigations of microbial infections greatly benefit from the increased information gained by whole-genome sequencing (WGS) in combination with standardized approaches for data sharing and analysis.Aim. To evaluate the quality and accuracy of WGS data generated by different laboratories but analysed by joint pipelines to reach a national surveillance approach.Methods. A national methicillin-resistant Staphylococcus aureus (MRSA) collection of 20 strains was distributed to nine participating laboratories that performed in-house procedures for WGS. Raw data were shared and analysed by three pipelines: 1928 Diagnostics, JASEN (GMS pipeline) and CLC-Genomics Workbench. The outcomes were compared according to quality, correct strain identification and genetic distances.Results. One isolate contained intraspecies contamination and was excluded from further analysis. The mean sequencing depth varied between sites and technologies. However, all analysis methods identified 12 strains that belonged to one of five outbreak clusters. The cut-off definition was set to <10 allele differences for core genome multilocus sequence typing (cgMLST) and <20 genetic differences for SNP analysis in a pairwise comparison.Conclusions. MRSA isolates, which are whole genome sequenced by different laboratories and analysed using the same bioinformatic pipelines, yielded comparable results for outbreak clustering for both cgMLST and SNP, using the 1928 analysis pipeline. In this study, JASEN was best suited to analyse Illumina data and CLC to analyse within respective technology. In the future, real-time sharing of data and harmonized analysis within the Genomic Medicine Sweden consortium will further facilitate investigations of outbreaks and transmission routes.

A genomic variation map provides insights into potato evolution and key agronomic traits.

Hybrid potato breeding based on diploid inbred lines is transforming the way of genetic improvement of this staple food crop, which requires a deep understanding of potato domestication and differentiation. Here, we resequenced 314 diploid wild and landrace accessions to generate a variome map of 47,203,407 variants. Using the variome map, we discovered the reshaping of tuber transcriptome during potato domestication, characterized genome-wide differentiation between landrace groups Stenotomum and Phureja, and identified a jasmonic acid biosynthetic gene possibly affecting tuber dormancy period. Genome-wide association studies revealed a UDP-glycosyltransferase gene for biosynthesis of antinutritional steroidal glycoalkaloids (SGAs), and a Dehydration Responsive Element Binding (DREB) transcription factor conferring increased average tuber weight. In addition, genome similarity and group-specific SNP analyses indicated tetraploid potatoes originated from the diploid S. tuberosum group Stenotomum. These findings shed light on the evolutionary trajectory of potato domestication and improvement, providing a solid foundation for advancing hybrid potato breeding practices.

Animal Brucellosis in Egypt: Review on Evolution, Epidemiological Situation, Prevalent Brucella Strains, Genetic Diversity, and Assessment of Implemented National Control Measures.

Brucellosis is a neglected zoonotic disease that has a significant economic and public health impact, especially in endemic countries. This review delves deeply into brucellosis's current epidemiological situation and potential sources of livestock infection in Egypt during the last two decades. MLVA-16 and Whole Genome Sequencing based on core-genome SNP analyses confirm the presence of different B. abortus and B. melitensis outbreak strains, both older widely disseminated Brucella strains and newly introduced ones. Despite implementing the test-and-slaughter control strategy over forty years, the disease is still endemic, and different Brucella species circulate among several animal species. The raising of mixed animal species in the same households or farms, exposure to aborted animals, and lack of public awareness about brucellosis transmission are among the main risk factors for increasing livestock brucellosis prevalence in Egypt. Young animals' voluntary vaccination, lack of a nationwide animal identification system, and uncontrolled animal movement stand beyond the ineffectively applied control strategy and may be subdued by applying mass vaccination to decrease disease prevalence dramatically and target imported camels, domestic pigs, and dogs (housed and stray) in the national control surveillance. Increasing awareness through educational campaigns is compulsory to reduce brucellosis transmission risk to livestock/humans.

In silico MLVA Analysis of Brucella melitensis from Human and Livestock in Iran.

Brucellosis, a zoonotic disease caused by Brucella spp. globally, is of great significance not only to livestock but also to public health. The most significant of the twelve species is Brucella melitensis. This article is devoted to the endemic region of Iran and aims to uncover the molecular epidemiology of B. melitensis. Biotyping, AMOS-PCR, Bruce-ladder PCR, and the in silico method of MLVA were employed to test 40 B. melitensis isolates from humans, cows, sheep, goats, camels, and horses which are found in thirteen Iranian provinces throughout the years 2015 to 2022. The data from the MLVA-8 analysis showed that there were seven genotypes that could be identified, and the most commonly identified genotype was genotype 63. The data from the MLVA-10 analysis showed that there were seven genotypes, with genotype 213 being the most prevalent in Iran. The data from the MLVA-11 analysis showed that there were eight genotypes, with genotype 111 being the most prevalent in Iran. The MLVA and SNP analysis results showed that the bacteria were grouped into two main groups, known as the Eastern Mediterranean and American groups. Moreover, the outcomes from these analyses have added considerably to our understanding of the genetic/historical relationships among the isolates. Our study indicates a high prevalence of B. melitensis biovar 1 in Iran, accounting for 82.5% of the isolates. The study provides insight into such matters as the complex epidemiology of B. melitensis in Iran, suggesting different ways of transmission and sources of infection. This research points out the vital significance of the continuation of the surveillance and curation of B. melitensis in the diverse species of animals and humans. The simplicity and efficiency of MLVA-based molecular epidemiology offer information on the geographic distribution and genetic diversity of B. melitensis and, therefore, help in the devising of targeted strategies for the prevention of disease in animals.

Identification of Bacillus anthracis Strains from Animal Cases in Ethiopia and Genetic Characterization by Whole-Genome Sequencing.

Anthrax is a zoonotic disease characterized by rapid onset with usual fatal outcomes in livestock and wildlife. In Ethiopia, anthrax is a persistent disease; however, there are limited data on the isolation and molecular characterization of Bacillus anthracis strains. This study aimed to characterize B. anthracis isolated from animal anthrax outbreaks between 2019 and 2024, from different localities in Ethiopia. B. anthracis was identified using standard microbiology techniques and confirmed by real-time PCR. For the first time in Ethiopia, the genetic diversity of five Bacillus anthracis strains, isolated from dead cattle and goats, was investigated by Whole Genome Sequencing (WGS) and bioinformatics analyses. The five sequenced strains were compared to one Ethiopian B. anthracis genome and the other 29 B. anthracis genomes available in the global genetic databases to determine their phylogeny. The genomes of the strains were also analyzed to detect the presence of antimicrobial resistance and virulence genes. The whole genome SNP analysis showed that the Ethiopian B. anthracis strains were grouped in the A clade. Three strains (BA2, BA5, and BA6) belonged to the A.Br.034 subgroup (A.Br.005/006), and two strains (BA1 and BA4) belonged to the A.Br.161 (Heroin) clade of the Trans-Eurasian (TEA) group. The findings of this study will contribute to expanding the current understanding of the anthrax hotspots in Ethiopia, and the phylogenetic correlation and/or diversity of the circulating strains.

Diversification of bla OXA-48-harbouring plasmids among carbapenemase-producing Enterobacterales, 11 years after a large outbreak in a general hospital in the Netherlands.

Introduction. Genes encoding OXA-48-like carbapenem-hydrolyzing enzymes are often located on plasmids and are abundant among carbapenemase-producing Enterobacterales (CPE) worldwide. After a large bla OXA-48 plasmid-mediated outbreak in 2011, routine screening of patients at risk of CPE carriage on admission and every 7 days during hospitalization was implemented in a large hospital in the Netherlands. The objective of this study was to investigate the dynamics of the hospitals' 2011 outbreak-associated bla OXA-48 plasmid among CPE collected from 2011 to 2021.Methods. A selection of 86 bla OXA-48-carrying CPE isolates was made from 374 isolates collected over an 11-year study period. Species included Escherichia coli (Eco), Klebsiella pneumoniae (Kpn), Enterobacter cloacae complex (Ecl), Citrobacter freundii (Cfr), Citrobacter koseri (Cko) and Morganella morgani (Mmo). Short-read sequencing was combined with long-read sequencing for all isolates to reconstruct bla OXA-48-like plasmids and chromosomes of CPE. MASH, MOBsuite, ResFinder, PlasmidFinder and SNP analyses were performed to study diversity. pOXA-48 plasmids were compared to plasmid sequences that were sequenced for the Dutch CPE surveillance in the same time period.Results. In total for the 86 CPE, 2 failed genomic assemblies and 78 bla OXA-48-encoding plasmids were reconstructed, and six bla OXA-48 genes were located chromosomally. The 2011 outbreak-associated bla OXA-48 plasmid of 63.6 kb with IncL replicon was found in Cfr, Ecl, Eco, Kpn and Mmo and primarily between 2011 and 2014 and indicated as LR025105 as MASH nearest neighbour. From 2014 onwards, 11 other types of bla OXA-48-carrying plasmids with different antibiotic-resistant genes and replicons were discovered, representing the earlier defined distinct pOXA-48 plasmid groups found in the Netherlands. Furthermore, on a national level, the LR025105 plasmid was found after 2015 in many different bacterial backgrounds, highlighting the promiscuous nature of this pOXA-48 plasmid.Conclusion. After a large bla OXA-48 outbreak in a large hospital in the Netherlands, the composition of the bla OXA-48 plasmid population in this hospital diversified over time and is in line with national surveillance data. Plasmid sequencing provided valuable insight into the transmission dynamics of bla OXA-48-encoding plasmids and showed no indication of the persistence of the 2011 bla OXA-48 plasmid in the hospital environment.

Genomic epidemiology of extended-spectrum beta-lactamase-producing Escherichia coli from humans and a river in Aotearoa New Zealand.

In Aotearoa New Zealand, urinary tract infections in humans are commonly caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli. This group of antimicrobial-resistant bacteria are often multidrug resistant. However, there is limited information on ESBL-producing E. coli found in the environment and their link with human clinical isolates. In this study, we examined the genetic relationship between environmental and human clinical ESBL-producing E. coli and isolates collected in parallel within the same area over 14 months. Environmental samples were collected from treated effluent, stormwater and multiple locations along an Aotearoa New Zealand river. Treated effluent, stormwater and river water sourced downstream of the treated effluent outlet were the main samples that were positive for ESBL-producing E. coli (7/14 samples, 50.0%; 3/6 samples, 50%; and 15/28 samples, 54%, respectively). Whole-genome sequence comparison was carried out on 307 human clinical and 45 environmental ESBL-producing E. coli isolates. Sequence type 131 was dominant for both clinical (147/307, 47.9%) and environmental isolates (11/45, 24.4%). Only one ESBL gene was detected in each isolate. Among the clinical isolates, the most prevalent ESBL genes were bla CTX-M-27 (134/307, 43.6%) and bla CTX-M-15 (134/307, 43.6%). Among the environmental isolates, bla CTX-M-15 (28/45, 62.2%) was the most prevalent gene. A core SNP analysis of these isolates suggested that some strains were shared between humans and the local river. These results highlight the importance of understanding different transmission pathways for the spread of ESBL-producing E. coli.

Investigation of Genetic Variants of Alox12 (rs9904779) Gene in Dental Caries

Background: Dental caries is a biofilm-mediated disease driven by dietary sugars that reduce pH and promote cariogenic microorganisms. Caries result from a complex interplay of behaviour, environmental, genetic, and physiological factors, with the immune response and bacterial activity contributing to enamel demineralization and cavity formation. Genetic factors, such as SNPs, also influence caries susceptibility, impacting enamel hardness and inflammatory responses. Aim: This study aims to explore the association between the Alox12 gene variant, rs9904779, and the susceptibility to dental caries. Methods: Patients were recruited following ethical approval and informed consent. Saliva samples were collected and grouped by DMFT index, and genomic DNA was extracted. PCR analysis focused on Alox12 gene polymorphism with the 223bp product and RFLP. Statistical analysis was performed using Epi Info software, calculating genotype-risk associations and a significance threshold of p &lt; 0.05. Chi-square testing assessed genotype and allele distributions across groups. Results: An SNP analysis of Alox12 was conducted to assess the susceptibility of dental caries. In caries patients, CC genotype was most prevalent (42%), while CG was higher in controls (52%). Genotype distribution deviated from Hardy-Weinberg equilibrium in caries (p &lt; 0.05) but not in controls (p &gt; 0.05). The caries group had a higher prevalence of the C allele, while the CG heterozygote was more frequent in controls (OR = 0.64, p = 0.32). Conclusion: This SNP analysis suggests that the Alox12 gene variant rs9904779 may be a significant predictor for the development of dental caries, highlighting its potential as a genetic marker for susceptibility to the disease.

Genome-Wide Patterns of Homozygosity and Heterozygosity and Candidate Genes in Greek Insular and Mainland Native Goats.

Runs of homozygosity (ROHs) and heterozygosity (ROHets) serve for the identification of genomic regions as candidates of selection, local adaptation, and population history. The present study aimed to comprehensively explore the ROH and ROHet patterns and hotspots in Greek native dairy goats, Eghoria and Skopelos, genotyped with the Illumina Goat SNP50 BeadChip. SNP and functional enrichment analyses were conducted to further characterize hotspots and the candidate genes located within these genomic regions. Genetic relationships between and within breeds and inbreeding coefficients were also evaluated. Clear genetic differentiation and diversified management practices were depicted between the two native populations. The ROH and ROHet average genome coverage for Skopelos (65.35 and 35 Mb) and Eghoria (47.64 and 43 Mb) indicated differences in mainland and insular goats, with Skopelos showing more long ROH fragments, reflecting its geographic isolation and small population size. An ROH hotspot (CHR12: 43.59-44.61 Mb) detected in the Skopelos population has been also reported across European goats and co-localizes with a selection signal detected in the Egyptian Barki goats and sheep adapted to hot-arid conditions. A novel ROH hotspot (CHR18: 60.12-61.81 Mb), shared among the Greek breeds, harbors candidate genes enriched in biosynthesis, metabolism, and immune response. Two well-conserved ROHet islands were detected in Greek goats on chromosomes 1 and 18, with genes participating in development and embryogenesis. The Eghoria population showed the highest number of ROHet islands, potentially reflecting its adaptability to diverse environments. These findings offer new insights into the environmental adaptation and artificial selection in Greek goats and could be utilized in future breeding strategies for sustainable goat farming.

Genome-Wide Identification and Expression Analysis of NF-YA Gene Family in the Filling Stage of Wheat (Triticum aestivum L.).

The NF-YA gene family is a highly conserved transcription factor that plays a crucial role in regulating plant growth, development, and responses to various stresses. Despite extensive studies in multiple plants, there has been a dearth of focused and systematic analysis on NF-YA genes in wheat grains. In this study, we carried out a comprehensive bioinformatics analysis of the NF-YA gene family in wheat, using the latest genomic data from the Chinese Spring. A total of 19 TaNF-YA genes were identified. An analysis of conserved domains, phylogenetic relationships, and gene structure indicated a significant degree of conservation among TaNF-YAs. A gene collinearity analysis demonstrated that fragment duplication was the predominant mechanism driving the amplification of TaNF-YAs. Furthermore, cis-acting elements within the promoters of TaNF-YAs were found to be implicated in grain development. Subsequently, SNP analysis revealed the genetic variation in the NF-YA gene family in different wheat. Moreover, published RNA-seq data were used and RNA-seqs of Pinyu8155, Yaomai30, Yaomai36, and Pinyu8175 were performed to identify TaNF-YAs influencing grain development. Finally, it was found that NF-YAs had no self-activating activity in wheat. This study provides key candidate genes for the exploration of grain development in the wheat filling stage and also lays a foundation for further research on the regulation of starch and protein synthesis and accumulation.

Comparative plastomic analysis of cultivated Dioscorea polystachya and its close relatives provides insights on the inter- and intraspecific phylogenies and potential wild origins of domestication

BackgroundDioscorea polystachya and its closely related species are original plants of the tuber crop “yam”, which had been intensively use for medicinal and food purposes and widely cultivated in northern China and its surrounding areas with a long history. Many cultivars of these species are often confused with one another because of similar tuber morphology, however, conventional DNA barcoding faces practical limitations restricting the method to effectively identify closely related species. In addition, phylogenetic relationships among various cultivar groups of Chinese yam (D. polystachya) remains unclear. To solve these problems, genomic DNAs of 15 Dioscorea samples were sequenced to assemble and annotate chloroplast genomes, which were used for analyzing their structural characteristics and identifying phylogenetic relationships at the inter- and intraspecific levels.ResultsThe size of chloroplast genomes of the tested samples is about 153 kb, and 79 protein-coding genes, 29 tRNA genes, and 4 rRNA genes are annotated. Phylogenetic analysis showed that D. polystachya were sister to Dioscorea japonica, and for Huaishan yams, Dioscorea persimilis did not cluster with Dioscorea alata and Dioscorea fordii. Four cultivar groups of Chinese yam were determined, namely Tiegun group, Anping group, Foshou group and Taihang complex group. Among these cultivar groups, Foshou and Taihang complex are clustered with different wild yams, respectively. Amino acid preferences are similar at the inter- and intraspecific levels, while synonymous codon usage reflects distinct patterns in the majority of cultivars of D. polystachya. There are distinct SSR variations among species, as well as four cultivar groups. Collinearity and SNP analyses show that nucleotide hypervariable regions among Dioscorea species are mainly concentrated in trnK–atpA, rps16–trnQ, atpA–atpH, rpoB–psbD, atpH–atpI, trnV–ndhC in the LSC region, and ccsA–ndhF in the SSC region, while intraspecific variation of Chinese yam is enriched in the intergenic spacers of rpoB–psbC, ndhD–ndhF, and trnQ-trnS, as well as the gene ycf1.ConclusionPhylogenetic analysis supports that Huaishan yams are not of monophyletic origin and the cultivated Chinese yam has at least two wild origins of domestication, which is consistent with the historical records of these wild yams from Mt. Dabie and Mt. Taihang. The identification efficiency of the newly developed barcodes for cultivar groups based on chloroplast genome SNP screening is significantly better than those of conventional barcodes. This approach to generate viable candidate markers based on the comparison from interspecific and intraspecific hypervariable regions of chloroplast genomes can be applied to conduct phylogenetic relationships of more important crop species and their close relatives, which are difficult to identify, as well as their wild origins of domestication.

The role of SNP (rs11908352) genetic polymorphisms on serum level of matrix metalloproteinase-9 in rheumatoid arthritis

Background. MMP-9 plays a significant role in the destruction of the extracellular matrix (ECM) in various physiological and pathological processes, including tissue remodeling. MMP-9 is elevated in pathologies involving abnormal immune responses. The extent of joint damage varies significantly among individuals with rheumatoid arthritis (RA), and much of its genetic inheritance remains unexplained. Multiple autoimmune diseases share common genetic risk factors that may also influence disease progression. Genetic variants in MMP-9 could potentially affect its transcriptional expression or enzymatic activity. Objective. To evaluate the impact of serum MMP-9 levels and the SNP (rs11908352) in a group of patients with rheumatoid arthritis compared to a control group. Methods. Seventy-three RA patients with a mean age of 47.91 ± 1.44 years were included in this study, compared to sixty-eight healthy controls matched in age and gender, with a mean age of 46.42 ± 1.51 years. Serum levels of MMP-9 and rs11908352 SNPs were examined in individuals with RA and compared to the control group. Results. The findings revealed a notable increase in MMP-9 levels in RA patients compared to healthy controls (1.85 ± 0.12 vs. 1.31 ± 0.10). Additionally, rs11908352 SNP analysis showed a statistically insignificant rise in the frequency of the AA genotype and A allele in the patient group compared to controls (24.66% vs. 16.18%; p = 0.297; OR = 1.70) and (51.0% vs. 44.0%; p = 0.285; OR = 1.30), respectively. Genotype frequencies CA, CC, and allele frequency C showed nonsignificant decreases in RA patients compared to controls (52.05% vs. 55.88%; p = 0.736; OR = 0.86), (23.29% vs. 27.94%; p = 0.566; OR = 0.78), and (49.0% vs. 56.0%; p = 0.285; OR = 0.77), respectively. Patients with the AA genotype exhibited markedly elevated serum MMP-9 levels compared to the control group (p = 0.007). Conclusion. The results of this study demonstrated a significant correlation between serum MMP-9 levels and rs11908352 genetic variation in RA patients, suggesting that specific genetic regions associated with autoimmune diseases may influence the severity of joint damage in rheumatoid arthritis.

BIOLOGICAL NOMENCLATURE (TAXONOMY) AND CLASSIFICATION OF HONEY BEE. CURRENT STATE AND PROBLEMS

The chronology of scientific research achievements in the process of determining the biological nomenclature and classification of honey bees and the implementation of the results obtained for use in the practice of beekeeping in the selection and preservation of aboriginal breeds (races) of bees in modern environmental conditions is highlighted. An attempt is made to focus this study in the field of analysis of the chronology of methods used in the historical aspect of determining the taxonomy of insects, in particular bees. At the same time, the problems and prospects of scientific research in modern economic and natural and climatic conditions of the development of the industry are reflected. It is shown that initially only morphometry was used worldwide to identify bee breeds. However, morphometric features are not always informative in identifying subspecies, since they are subject to variability under the influence of environmental conditions. Later, biochemical methods for identifying bee subspecies based on polymorphism of allozyme loci were developed. It is shown that at the same time, methods for identifying bee subspecies based on polymorphism of mitochondrial DNA (mtDNA) loci were developed. This polymorphism was successfully used in phylogenetic and phylogeographic studies of honey bees. The disadvantage of mtDNA markers is the exclusively maternal type of inheritance. At the same time, methods for identifying bee subspecies were developed taking into account polymorphism of nuclear DNA (nDNA) loci. Recently, methods for identifying bee subspecies based on SNP analysis have been developed. These markers have become widely used in population, evolutionary and phylogenetic studies of bees due to the development of next-generation sequencing methods NGS (Next Generation Sequencing) Illumina. SNP markers are characterized by high resolution due to their number and stable inheritance over several generations, which can be successfully used in genetic mapping, population and evolutionary studies, selection of lines for economically useful traits and disease resistance, identification of taxonomic affiliation of bee families.

The two-component sensor factor envZ influences antibiotic resistance and virulence in the evolutionary dynamics of multidrug-resistant Salmonella enteritidis causing multisite invasive infections.

To assess the impact of mutations in the two-component sensor envZ on antibiotic resistance and virulence in the evolutionary dynamics of MDR Salmonella enteritidis (S. enteritidis). Five S. enteritidis isolates obtained from a patient with multisite invasive infections were analysed. Analysis of antibiotic resistance genes, virulence genes and SNP was performed through WGS. RNA sequencing, quantitative RT-PCR, virulence testing in a Galleria mellonella (G. mellonella) infection model and in vitro cell experiments were used to examine the effects of envZ mutations. WGS revealed identical resistance and virulence genes on an IncFIB(S)/IncFII(S)/IncX1 fusion plasmid in all strains. The faecal strains harboured envZ mutations, reducing outer membrane protein ompD and ompF transcriptional level. Virulence testing demonstrated elevated virulence in envZ mutant strains. In vitro experiments revealed increased adhesion, invasion and phagocytosis resistance in envZ mutants, along with reduced biofilm formation and growth rates. These findings highlight novel genetic locations on envZ influencing antibiotic resistance and virulence in clinical S. enteritidis strains. envZ mutations impact antibiotic resistance by down-regulating ompD and ompF expression and enhance virulence, contributing to multisite infections with increased fitness costs.

Differential vasoproliferative traits of Bartonella henselae strains associated with autotransporter BafA variants.

Bartonella henselae, a Gram-negative facultative intracellular bacterium, is the etiological agent of cat-scratch disease and also causes bacillary angiomatosis in immunocompromised individuals. Although the ability to promote vascular endothelial cell proliferation differs among Bartonella species, variations among strains within B. henselae remain unclear. Bartonella angiogenic factor A (BafA) and Bartonella adhesin A (BadA) have been identified as autotransporters of B. henselae that are involved in endothelial cell proliferation. Although strain-specific differences in the expression of BadA and the VirB/D4 type IV secretion system have been reported, BafA expression among B. henselae strains has yet to be examined. Therefore, the present study investigated the proliferation-promoting ability of 13 B. henselae strains from several sources in human umbilical vein endothelial cells (HUVECs). We identified BafA variants 1 and 2 based on the deduced amino acid sequences of its passenger domain. The recombinant proteins of both variants exhibited similar proliferation activity against HUVECs. However, BafA variant 2 strains showed cytotoxicity at a high bacterial inoculum in a direct coculture with HUVECs, which was attenuated in an indirect coculture. These strains, in contrast to BafA variant 1 strains, highly expressed BadA and exhibited bacterial aggregation. Based on a core genome SNP analysis of 50 B. henselae strains, the BafA variant types corresponded to clades 1-4. These results indicate that vasoproliferative traits differ among B. henselae clades based on the variant types. Therefore, this study provides a new conceptual framework in which the clades of B. henselae may predict their pathogenicity in humans.IMPORTANCEBartonella species including Bartonella henselae, Bartonella quintana, and Bartonella bacilliformis cause vasoproliferative lesions. Their proliferation-promoting ability in vascular endothelial cells differs among Bartonella species; however, it is unclear whether these differences exist among B. henselae strains. We herein showed that B. henselae strains exhibited variable proliferation-promoting ability and cytotoxicity in vascular endothelial cells, which corresponded to the bafA gene variants possessed by the strains. The expression levels of Bartonella angiogenic factor A (BafA) and Bartonella adhesin A, as well as the degree of proliferation-promoting ability and cytotoxicity in endothelial cells, varied among the strains. A core genome SNP analysis of strains using whole genome sequencing data divided B. henselae strains into four clades, with each clade corresponding to BafA variants 1-4. These results suggest the differential vasoproliferative potency of B. henselae, with potential implications in clinical management, including risk stratification and predictions of the clinical course.

SNP Fingerprinting for Germplasm Identification of the Fast-Growing Pacific oyster (Crassostrea gigas) "Haida No. 1" Variety.

The Pacific oyster (Crassostrea gigas) is a global aquaculture species of economic significance. Selective breeding programs have been conducted to produce multiple strains with fast growth as well as other desirable traits. However, due to the phenotypic plasticity of oysters, challenges existed for precise germplasm identification among selectively bred strains. In this work, we identified selection signatures of three fast-growing Pacific oyster strains originated from wild populations collected from China, Japan, and Korea, respectively, which were used for development of SNP-based molecular fingerprinting for precise identification of germplasm. We performed whole-genome resequencing of 59 oysters from three selectively bred strains and a wild population for genome-wide SNP analyses. Population structure analysis with these SNPs revealed significant genetic differentiation among the selectively bred strains. Based on the FST index, we identified 41, 49, and 36 strain-specific SNPs from the three selectively bred strains. Taking into account the "hitch-hiking effect" that occurs in the genome during positive selection, we identified two, three, and two molecular fingerprints for the three strains, respectively. We validated the molecular fingerprints of the China selectively bred strain (i.e., "Haida No. 1" variety) with a separate population of 42 oysters with diverse genetic background, demonstrating the accuracy of germplasm identification of over 96%. This work provides a reliable tool for precise germplasm identification of the "Haida No. 1" variety as well as other two selectively bred strains, which is valuable in germplasm conservation and breeding design in the C. gigas.

Homologous recombination deficiency in pancreatic neuroendocrine tumors.

Aim: Pancreatic Neuroendocrine tumors (pNETs) are a heterogeneous group of neoplasms whose tumor biology is still little known. Thanks to next-generation sequencing, pathogenic mutations in base-excision-repair MUTYH gene and homologous recombination genes CHEK2 and BRCA2 seem to have a role in the development of pNETs.Research design & methods: We assumed that Homologous Recombination Deficiency (HRD) could be a critical pathogenetic mechanism for pNETs. We evaluated the HR status in a case series of 33 patients diagnosed with pNET at the Modena Cancer Center using the AmoyDX HRD Focus assay.Results: The AmoyDx test did not identify any HRD-positive patients (median GSS equal to 1.1, positive score: >50), and no pathogenic BRCA variants were detected. However, thanks to the SNP analysis, a consistent number of partial or complete single-copy deletions or duplications in several chromosomes.Conclusion: The AmoyDX HRD focus assay performed well on pancreatic samples, despite being originally designed for ovarian cancer and used on samples stored for over a year. Larger studies are needed to further assess the role of HRD assays in pNETs research.

Genetic characterization and clonal analysis of carbapenemase-producing Escherichia coli and Klebsiella pneumoniae from canine and human origins.

Carbapenem-resistant Enterobacterales (CRE), particularly carbapenemase-producing Escherichia coli and Klebsiella pneumoniae, pose a significant global health challenge due to their resistance to last-resort antibiotics. This study investigates the genetic characteristics and clonal relationships of CRE isolated from canine and human clinical samples in Bangkok to understand potential interspecies transmission. Fifty-two CRE isolates were collected from 477 clinical samples from dogs and humans at Chulalongkorn University between 2017-2021. Bacterial species were identified using MALDI-TOF, and antimicrobial resistance was confirmed through broth microdilution testing. Genetic analyses included plasmid replicon typing, multilocus sequence typing (MLST), whole genome sequencing (WGS), and pulsed-field gel electrophoresis (PFGE) to assess resistance genes and clonal relatedness. CRE isolates from both species exhibited genetic variability with high ARG counts, particularly in human isolates. MLST identified ST410 in most E. coli isolates from both dogs and humans, and IncFIA/IncFIB plasmids were predominant among blaNDM-positive isolates. PFGE patterns and SNP analysis showed no clonal relationship between canine and human isolates, suggesting independent acquisition pathways for CRE in the two hosts. The study highlights the absence of direct clonal transmission between canine and human isolates but reveals overlapping sequence types and plasmid types. The findings underscore the potential for interspecies transmission under certain conditions, emphasizing the importance of a One Health approach for monitoring CRE in both human and animal populations.

Vitamin D receptor gene polymorphisms in patients with relapsing multiple sclerosis.

The relationship between the vitamin D receptor (VDR) gene and many pathogenic pathways in relapsing-remitting multiple sclerosis (RRMS) remains unclear. Given the significance of the topic, we conducted this study to explore the correlation between vitamin D receptor gene polymorphisms and clinical and inflammatory factors in patients suffering from relapsing-remitting multiple sclerosis. The current research is a case/control study conducted based on the Helsinki Ethical Principles. RRMS disease was confirmed based on history, clinical symptoms, radiological signs and neurologist diagnosis. The research population consisted of healthy people and patients with RRMS who were referred to Hazrat Rasool Akram Hospital between 2021 and 2023. For each person participating in the study (RRMS patient and healthy), five milliliters of peripheral blood containing the anticoagulant EDTA was collected. Polymerase chain reaction was performed using two specific and appropriate oligonucleotide primers. The restriction fragment length polymorphism technique was used, one of the standard methods for identifying polymorphisms. Statistical analysis was performed using SPSS software version 23. The odds ratio and 95% confidence limits were calculated. The SNP Analyzer software was used to analyze the allele frequency of each polymorphism in healthy and RRMS individuals and compare the values. Prism version 5 software was used to draw diagrams. In the present study, a statistically significant difference was observed between the percentage of FokI genotypes in RRMS patients and healthy individuals. FokI polymorphism showed a significantly increased risk with an odds ratio of 7.28 in patients with the FF genotype compared to healthy individuals. ApaI, TaqI, and BsmI were not significantly different between the two groups. Based on the findings of the present study, FokI polymorphism showed a significant risk increase in RRMS patients with FF genotype compared to healthy individuals.

Banded or striped? Significant colour dimorphism of a bridal snake in Borneo

AbstractStripes and bands are common colour patterns in snakes. Some species are known to exhibit colour polymorphism and that includes expressing it in the bands and stripes. Bridal snake, Dryocalamus is a medium‐sized arboreal snake in South to Southeast Asia and most species of the genus are banded, but including two striped species. In Borneo, banded D. subannulatus and striped D. tristrigatus occur sympatrically and the two species are similar morphologically except for their colour patterns. We hypothesized the two species in Borneo exhibit a significant colour dimorphism of one species and conducted morphological and genetical analysis to test the hypothesis. Morphological examinations revealed that D. subannulatus and D. tristrigatus are not distinguishable in Borneo by scalation. Mitochondrial phylogenetic analysis and nuclear SNP analysis showed D. subannulatus and D. tristrigatus from Borneo formed clades or clusters by region not by species. Dryocalamus subannulatus and D. tristrigatus in Borneo are morphologically and genetically indistinguishable and Bornean D. subannulatus should be treated as a colour morph of D. tristrigatus. This is a rare example of sympatrically distinct banded/striped dimorphism in snakes. Relative abundance of banded/striped morph of the genus is probably different depending on region, and any ecological factor may contribute to it.