Genotypic Characteristics Research Articles

Phenotypic and Genotypic Characterization of Candida parapsilosis complex isolates from a Lebanese Hospital.

The opportunistic fungal pathogen Candida parapsilosis is a major causative agent of candidiasis leading to death in immunocompromised individuals. Azoles are the first line of defense in treatment by inhibiting ERG11, involved in the synthesis of ergosterol, the main sterol fungal sterol. Resistance to azoles is on the increase worldwide including in Lebanon. The purpose of this study is to characterize nine hospital isolates labeled as C. parapsilosis: four resistant and five sensitive to fluconazole. Phenotypic characterization was achieved through a battery of tests that target pathogenicity attributes such as virulence, biofilm formation, stress resistance, and ergosterol content. Genotypic analysis was done through whole genome sequencing to mutations in key virulence and resistance genes. Phylogenetic comparison was performed to determine strain relatedness and clonality. Genomic data and phylogenetic analysis revealed that three of the nine C. parapsilosis isolates were misidentified; two as C. orthopsilosis and C. metapsilosis belonging to the C. parapsilosis complex, while the third was C. albicans. Moreover, several known and novel mutations in key drug resistance and virulence genes were identified such as ERG11, ERG3, ERG6, CDR1, and FAS2. Phylogenetic analysis revealed a high degree of relatedness and clonality within our C. parapsilosis isolates. Our results showed that resistant isolates had no increased ergosterol content, no statistically significant difference in virulence, but exhibited an increase in biofilm content compared to the sensitive isolates. In conclusion, our study, the first of its kind in Lebanon, suggests several mechanisms of antifungal drug resistance in C. parapsilosis hospital isolates.

Streptomyces chengbuensis sp. nov., isolated from the rhizosphere soil of Cathaya argyrophylla.

Strain HUAS CB01T was a novel actinobacterium which was isolated from the rhizosphere soil of Cathaya argyrophylla, Chengbu Miao Autonomous County of Hunan Province, China. The strain formed well-growing substrate mycelium, diffusible pigments, and aerial mycelium, and differentiated into spiral-type spore chains composed of smooth-surface rod-shaped spores. Phylogenetic analysis on account of 16 S rRNA gene sequence demonstrated the strain HUAS CB01T was a member of the genus Streptomyces and had a close relationship with Streptomyces wuyuanensis CGMCC 4.7042 T (100%) and Streptomyces marianii ICN19T (99.86%). Genome-based comparisons indicated that strain HUAS CB01T could be distinctly different from its closest species, Streptomyces wuyuanensis CGMCC 4.7042 T, Streptomyces marianii ICN19T, with ANIm and dDDH results of 92.78% and 45.90%, 92.22% and 43.30%, respectively, far less than 96.7 and 70% cut-off points recommended for delineating species. The main cellular fatty acids concluded anteiso-C15:0, iso-C14:0, iso-C16:0, C16:0 and C16:1 2OH. The menaquinones were MK-9(H4), MK-9(H6) and MK-9(H8) and the whole-cell sugars consisted of ribose and mannose. The polar lipids included phosphatidyl ethanolamine, diphosphatidylglycerol, phosphatidylglycerol, mannosides and unidentified phospholipids. According to these genotypic and phenotypic characteristics, strain HUAS CB01T can be distinguished and representative to be a novel species of the genus Streptomyces, for which the name Streptomyces chengbuensis is proposed. The type strain is HUAS CB01T ( = MCCC 1K08666T = JCM 36277 T).

Epidemiological study of antibiotic-resistant Helicobacter pylori in Brazil a meta-analysis

Objectives: To evaluate the proportion of Helicobacter pylori resistant to tetracyclines, amoxicilline, furazolidone, clarithromycin, fluoroquinolones and amoxicillin in Brazil and to present genotypic characteristics of strains. Methodology: Search in PubMed, SciELO, VHL and Medline. Twelve Brazilian studies on resistant Helicobacter pylori were selected, with a total of 2,038 patients. The options were performed with a confidence interval of 95% and an alpha of 0.05 using the software RStudio5.0. Results: low resistance rate for 04 antibiotics, being lower than the 15% threshold considered for empirical treatment. However, for metronidazole the rate of 43% and clarithromycin 16% macrolide and amoxicillin increased resistance when compared to previous years. The Southeast region had the highest rates. A clarithromycin ae A2147 in the 23s 23s RNA gene was the most prevalent conferring clear resistance. Conclusion: The increase in resistance to clarithromycin and amoxicillin resistance demonstrates that the factors that still increase for the acquisition of resistance still exist. Therefore, constant epidemiological surveillance is necessary so that we are aware of the resistance profile, performing assertive therapy.

Phenotypic and Genotypic Features of the FAN1 Mutation-Related Disease in a Large Hungarian Family.

Pathogenic variants in the FAN1 gene lead to a systemic disease with karyomegalic interstitial nephritis (KIN) at the forefront clinically. The phenotypic-genotypic features of a FAN1 mutation-related disease involving five members of a Hungarian Caucasian family are presented. Each had adult-onset chronic kidney disease of unknown cause treated with renal replacement therapy and elevated liver enzymes. Short stature, emaciation, latte-colored skin, freckles, and a hawk-like nose in four patients, a limited intellect in two patients, and chronic restrictive lung disease in one patient completed the phenotype. Severe infections occurred in four patients. All five patients had ceased. Four patients underwent autopsy. KIN and extrarenal karyomegaly were observed histologically; the livers showed no specific abnormality. The genotyping using formalin-fixed tissue samples detected a hitherto undescribed homozygous FAN1 mutation (c.1673_1674insT/p.Met558lfs*4; exon 5) in three of these patients and a heterozygous FAN1 mutation in one patient. The reason for the heterozygosity is discussed. In addition, 56 family members consented to the screening for FAN1 mutation from which 17 individuals proved to be heterozygous carriers; a blood chemistry evaluation of their kidney and liver function did not find any abnormality. The clinical presentation of FAN1-related disease was multifaceted, and not yet described manifestations were observed besides kidney and liver disease. Mutation in this gene should be suspected in adults with small kidneys of unknown cause, elevated liver enzymes, and recurrent infections, even without a family history.

Phenotypic and genotypic characterization of Staphylococcus aureus strains isolated from otitis externa: Emergence of CC30-spa t019-SCCmec IV carrying PVL as major genotype

The increasing emergence of Staphylococcus aureus as the primary causative agent of otitis externa has been noted; however, detailed information regarding the molecular characteristics of these strains in Iran remains scarce. The current study aims to investigate both genotypic and phenotypic attributes of S. aureus strains implicated in ear infections. In the present work, we analyzed 60 S. aureus strains isolated from cases of otitis externa over a period of 45 months. The resistance patterns were determined using disk diffusion and microbroth dilution methods. All S. aureus isolates were confirmed by the nucA polymerase chain reaction assay, and their biofilm production was assessed by a microtiter plate assay. Molecular characterization of the isolates was performed using the staphylococcal cassette chromosome mec, multilocus sequence typing, and staphylococcus protein A typing methods. Overall, the results indicated that 44 out of 60 S. aureus isolates (73.3 %) were methicillin-resistant Staphylococcus aureus. Resistance to mupirocin and vancomycin was observed in 13.3 % and 1.7 % of the tested isolates, respectively. Furthermore, out of the 60 S. aureus isolates, 56 strains (93.4 %) were classified as positive biofilm strains at different levels. Twelve distinct clonal lineages were identified. The vast majority of S. aureus isolates belonged to CC30/ST30-MRSA IV/t019 (41.7 %). Among the 31 strong biofilm producers, the majority (64.5 %) belonged to CC30/ST30-MRSA IV/t019 clone. Biofilm negative isolates belonged to CC22/ST22 (2 isolates), CC8/ST585 (one isolate), and CC8/ST8 (one isolate). Our result revealed that about three-quarters of PVL-positive strains belonged to CC30/ST30. Our data confirmed the presence of MSSA strains among CC30/ST30 and CC22/ST22 isolates. The mupirocin resistant isolates (n = 8) belonged to CC8/ST585-MRSA III/t713 (37.5 %), CC8/ST239-MRSA III/t030 (25 %), CC8/ST8-MRSA IV/t008 (12.5 %), CC8/ST239-MRSA III/t037 (12.5 %), and CC22/ST22-MRSA IV/t790 (12.5 %) lineages. The VRSA strain belonged to the CC8/ST8-MRSA IV/t008 lineage, carrying the vanA determinant. iMLSB phenotypes (n = 14) were distributed across different lineages, including CC30/ST30-MRSA IV/t019 (21.5 %), CC30/ST30-MSSA/t021 (21.5 %), CC22/ST22-MSSA/t005 (14.3 %), CC8/ST239-MRSA III/t030 (14.3 %), CC22/ST22-MSSA/t1869 (7.1 %), CC22/ST22-MRSA IV/t790 (7.1 %), CC8/ST239-MRSA III/t037 (7.1 %), and CC1/ST772-MRSA IV/t10795 (7.1 %). These findings highlight significant genotypic diversity and high biofilm formation among our isolates. The frequent occurrence of the CC/ST30 clone in S. aureus strains isolated from otitis externa reflects the emergence of these lineages as a predominant clone in Iran, posing a significant public health concern.

Altererythrobacter litoralis sp. nov., a New Carotenoid-Producing Member of the Family Erythrobacteraceae, Isolated from a Tidal Flat Sediment.

A new isolate designated as 1XM1-14T was isolated from a tidal flat sediment of Xiamen Island. The yellow-pigmented colonies and rod-shaped cells were observed. Strain 1XM1-14T could hydrolyze Tweens 20, 40, 60, aesculin, and skim milk, and was chemoheterotrophic and mesophilic, required NaCl for the growth. The 16S rRNA gene-based phylogenetic analysis indicated that strain 1XM1-14T was the most closely related to Altererythrobacter epoxidivorans CGMCC 1.7731T (97.0%), followed by other type strain of the genus Altererythrobacter with identities below 97.0%. The DNA-DNA hybridization and average nucleotide identity values between strain 1XM1-14T and its relatives of the genus Altererythrobacter were below the respective thresholds for prokaryotic species demarcation. The phylogenomic inference further revealed that strain 1XM1-14T formed a separate branch distinct from the type strains of the recognized species within the genus Altererythrobacter. The major cellular fatty acids of strain 1XM1-14T were identified as summed feature 8 (C18:1 ω6c and/or C18:1 ω7c), C17:1 ω6c, and C16:0; the profile of polar lipids comprised diphosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, an unidentified glycolipid, and two unidentified lipids; the respiratory quinone was determined to ubiquinone-10. The genomic size and DNA G+C content of strain 1XM1-14T were 2.5 Mbp and 62.71%. The key carotenoid biosynthetic genes were determined in the genome of strain 1XM1-14T and the generated carotenoids were detected. The combined genotypic and phenotypic characteristics supported the classification of strain 1XM1-14T (= GDMCC 1.2383T = KCTC 82612T) as a novel species in the genus Altererythrobacter, for which the name Altererythrobacter litoralis sp. nov. is proposed.

Phenotypic and genotypic characterization of methicillin and vancomycin-resistant Staphylococcus aureus isolated from human clinical samples in Kerman hospitals, Iran

Today, methicillin-resistant S. aureus (MRSA) isolates are widely spread in both communities and hospitals, causing high morbidity and mortality worldwide. However, the emergence of vancomycin resistance among MRSA isolates has been perceived as a formidable threat to therapeutic management. Therefore, the present study aimed to determine the frequency and antibiotic susceptibility of MRSA and vancomycin-resistant S. aureus (VRSA) strains in Kerman hospitals in Iran. A total of 455 human clinical samples were collected from Kerman hospitals. The samples were examined for the presence of S. aureus and the isolates were tested for their susceptibility to antibiotics by disk diffusion method. MRSA and VRSA isolates were detected by a combination of phenotypic and genotypic methods. Sixty-five coagulase-positive S. aureus strains were isolated, and the higher isolation rate was from wound samples (47.7%) compared with blood (18.5%), respiratory tract secretions (10.8%), urine (10.8%), body fluids (9.2%), abscess (1.5%), and cerebrospinal fluids (1.5%). The isolates exhibited high resistance toward tetracycline (69.2%). Thirty-four isolates (52.3%) were categorized as MRSA (phenotypically resistant to cefoxitin). mecA gene was amplified in 97.1% of MRSA strains. Moreover, only one MRSA strain was resistant to vancomycin (MIC=128), and this isolate carried the vanA gene. The frequency of MRSA was not significantly associated with gender, age, sample type, and sampling locality (p>0.05). Given the alarming rate of resistance among MRSA isolates, monitoring of antibiotic resistance should be performed to reduce treatment failure in patients with staphylococcal infections. Although vancomycin remains a drug of choice for MRSA, our study suggests that its efficacy may be limited by resistance development.

Prevalence, Genotypic Characteristics, and Antibiotic Resistance of Listeria monocytogenes From Retail Foods in Huzhou, China

Listeria monocytogenes are considered to be the major foodborne pathogen worldwide. To understand the prevalence and potential risk of L. monocytogenes in retail foods, a total of 1243 retail foods in 12 food categories were sampled and screened for L. monocytogenes from 2020 to 2022 in Huzhou, China. A total of 46 out of 1234 samples were confirmed to be L. monocytogenes positive with a total rate of 3.7%. The contamination rate of seasoned raw meat (15.2%) was the highest, followed by raw poultry meat and raw livestock meat (9.9%) and salmon sashimi (9.5%). The L. monocytogenes isolates belonged to four serotypes, 1/2a,1/2b, 1/2c, and 4b, with the most prevalent serotype being 1/2a (47.9%). All isolates were grouped into 15 sequence types (STs) belonging to 14 clonal complexes (CCs) via multilocus sequence typing (MLST). The most prevalent ST was ST9/CC9 (23.9%), followed by ST3/CC3 (19.6%) and ST121/CC121 (17.4%). Notably, 11 STs were detected from ready-to-eat (RTE) foods, some of them have been verified to be strongly associated with clinical origin listeriosis cases, such as ST3, ST2, ST5, ST8, and ST87. Listeria pathogenicity islands 1 (LIPI-1) and LIPI-2 were detected in approximately all L. monocytogenes isolates, whereas the distribution of both LIPI-3 genes and LIPI-4 genes exhibited association with specific ST, with LIPI-3 in ST3 and ST288, and LIPI-4 in ST87. The strains carrying LIPI-3 and LIPI-4 virulence genes in this study were all isolated from RTE foods. Antimicrobial susceptibility tests showed that >90% of isolates were susceptible to PEN, AMP, ERY, CIP, SXT, VAN, CHL, and GEN, indicating the antibiotic treatment might be still efficient for most of the L. monocytogenes strains. However, for the three clinical first-line antibiotics (PEN, AMP, and GEN), we also observed three and four strains showing MIC values greater than the susceptibility standards for PEN and AMP, respectively, and one strain showing resistance to GEN.

Morpho-biochemical Characterization of Tamarind (Tamarindus indica L.)

Background: The research was conducted at the ICAR-CRIDA Hayatnagar Research Farm in Hyderabad, India. The plant material consists of tamarind trees planted in 1998 with 5m spacing to improve the morphological and reproductive characteristics of elite genotypes as well as quality and biochemical characters among the twenty tamarind accessions maintained at the research farm. The experiment was started in 1998 and observations were taken over the fruiting season of 2021-2022 with twenty tamarind accessions; the experiments were established in a randomized block design. The trees were identified for their consistent health and development. Methods: Biometric observations mainly average number of flowers per inflorescence, average number of inflorescence per branch, average number of branches per tree, average fruit weight (g), average yield per plant (kg), fruit, pulp, seed weights as well as shell, fiber, fruit, number of normal seeds per pod as well as damaged number of seeds per pod, Carbohydrates g/ 100 g, Polyphenols g/100 g, Anthocyanin mg/100 g, % Antioxidant activity and % Tartaric acid were recorded and analyzed statistically. Result: Significant differences among the tamarind accessions evaluated, NZB(S), Hasanur #5, Salem 132, NTI-14 and SMG-3 recorded the highest values in all the growth, pod and yield characters. NZB(S) recorded the highest number of flowers per inflorescence (14.62) while Hasanur # 5 recorded the highest number of inflorescence per branch (13.87). In yield attributes, NZB(S) recorded the highest average yield per plant (kg) (15.72) followed by Hasanur #5 (15.09), Salem 132 (14.81) and NTI-14 (14.65). The results revealed that NZB(S) showed the highest mean performance in terms of growth, yield and quality characters. The best performing accessions are being multiplied through vegetative propagation methods for planting on large scale in different locations.

#174 Valvulopathies and left ventricular hypertrophy in ADPKD: prevalence and association with genotype and phenotype

Abstract Background and Aims Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited nephropathy due to mutations in PKD1 or PKD2 genes. In addition to kidney involvement, ADPKD is also characterized by extrarenal manifestations including the presence of cysts in other organs, intracranial aneurysms, arterial hypertension, valvulopathies and left ventricular hypertrophy (LVH). Cardiac abnormalities are an important cause of morbidity and mortality in ADPKD patients. Limited data on the association between these cardiovascular manifestations with phenotypic (both renal and systemic) and genotypic features of ADPKD are currently available. In this study we report the prevalence of the main cardiac abnormalities (valvulopathies, with a focus on aortic, mitral, tricuspid valve regurgitation, mitral prolapse, and LVH) observed in a large cohort of ADPKD patients, and investigated their association with renal diameter, renal function, other systemic manifestations such as pancreatic cysts, intracranial cysts, intracranial aneurysm and genotype. Method This retrospective study was conducted at the A. Gemelli Hospital, Rome in a single-center cohort of 129 ADPKD patients, who had previously undergone echocardiogram, abdominal ultrasound, brain MRI, and blood tests, between January 2018 and July 2023. Genetic testing was performed in 58 of the 129 patients. For the statistical analysis the average bilateral renal diameter of each patient was used. An interventricular septum cut-off thickness of 11 mm was used to define left ventricular hypertrophy. Non-parametric (Mann–Whitney and Spearman) and chi-square tests were applied as appropriate. Results In our cohort of 129 patients, the prevalence of valvulopathies was the following: 68% of mitral regurgitation, 22% of mitral prolapse, 20% of aortic regurgitation and 56% of tricuspid regurgitation. Left ventricular hypertrophy was identified in 28% of the entire cohort. We found that in ADPKD patients with aortic regurgitation, renal diameters are larger (p = 0.027) and the eGFR is lower (p < 0.01) than in patients without this echocardiographic abnormality (Fig. 1a,b). In the subset of genotyped ADPKD patients (58), mutations in PKD 1 were associated with mitral prolapse (p = 0.02). Interventricular septal thickness, regardless of the presence of hypertrophy was positively correlated (Fig. 1c) with renal diameter (p = 0.001 r = +0.289) and negatively (Fig. 2a) with eGFR (p < 0.001 r = −0.422). Both correlations, also adjusted for arterial hypertension, remained statistically significant (p = 0.008, coefficient r = +0.235 and p < 0.001, coefficient r = −0.304, respectively). In addition, patients with LVH featured a larger renal diameter (p = 0.03), and a lower eGFR (p < 0.001) (Fig. 2b-c). Moreover LVH was associated with the presence of PKD1 mutations (p = 0.03). Neither valvulopathies nor LVH were associated with other systemic manifestations of the disease. Conclusion We show for the first time an association of LVH with ADPKD genotype (specifically mutations in PKD1) and the association of a specific valvulopathy (aortic regurgitation) with the renal phenotype of ADPKD patients. To date only one study found an association of another valvulopathy, mitral regurgitation, with mutations in PKD1 [1]. Heart valves disease and LVH are risk factors for the subsequent development of further cardiovascular complications therefore understanding the pathogenetic mechanisms responsible for the higher incidence of these abnormalities in ADPKD would be useful to identify high-risk subgroups of patients and tailor personalised interventions.

Characterization of Actin-based Genotypes and Mycoplasma Endosymbionts of Trichomonas vaginalis Isolated in Sapporo, Japan.

Trichomonas vaginalis is a causative agent of common non-viral sexually transmitted infections worldwide. However, the biological features, such as genotypes and endosymbionts, of T. vaginalis isolated in Japan remain unclear. The aim of this study was to characterize the actin-based genotypes and the endosymbionts of T. vaginalis isolated in Sapporo, Japan. Three T. vaginalis clinical strains were isolated in Sapporo, Japan between 2019 and 2022. Actin-based genotyping was conducted by sequencing and phylogenetic analyses. The endosymbionts, such as Mycoplasma sp. and Trichomonasvirus, were detected using PCR and RT-PCR, respectively. Furthermore, the detected Mycoplasma spp. were identified using 16S rRNA gene sequencing. Of the three T. vaginalis strains, two belonged to genotype E, whereas one was genotype G as determined by actin-based genotyping. Two of the T. vaginalis strains harbored Mycoplasma spp. Using nearly full-length 16S rRNA gene sequencing, both were identified as Candidatus Mycoplasma girerdii. In contrast, the Trichomonasvirus was not found in the T. vaginalis strains. To our knowledge, this is the first report on the characterization of actin-based genotypes and the presence of endosymbiotic Ca. M. girerdii in T. vaginalis strains in Japan. Thus, this study will provide an important impetus for future research.

Clinical relevance of SCN and CyN induced by ELANE mutations: a systematic review.

According to the PRISMA criteria, a systematic review has been conducted to investigate the clinical relevance between patients with severe congenital neutropenia (SCN) and cyclic congenital neutropenia (CyN) induced by ELANE mutations. We have searched PubMed, EMBASE, Web of Science, Scopus, Cochrane, CNKI, Wanfang Medicine, and VIP for ELANE mutation related literature published from 1997 to 2022. Using Microsoft Excel collect and organize data, SPSS 25, GraphPad Prism 8.0.1, and Omap analyze and plot statistical. Compare the gender, age, geography, mutation sites, infection characteristics, treatment, and other factors of SCN and CyN patients induced by ELANE mutations, with a focus on exploring the relationship between genotype and clinical characteristics, genotype and prognosis. This study has included a total of 467 patients with SCN and 90 patients with CyN. The onset age of SCN and CyN are both less than 1 year old, and the onset and diagnosis age of SCN are both younger than CyN. The mutation of ELANE gene is mainly missense mutation, and hot spot mutations include S126L, P139L, G214R, c.597+1G>A. The high-frequency mutations with severe outcomes are A57V, L121H, L121P, c.597+1G>A, c.597+1G>T, S126L, C151Y, C151S, G214R, C223X. Respiratory tract, skin and mucosa are the most common infection sites, Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli are the most common. Patients with refractory G-CSF are more likely to develop severe outcomes. The commonly used pre-treatment schemes for transplantation are Bu-Cy-ATG and Flu-Bu-ATG. The prognosis of transplantation is mostly good, but the risk of GVHD is high. https://www.crd.york.ac.uk/PROSPERO/. PROSPERO, identifier CRD42023434656.

Comparative characterization of interleukin genotypes and risks of bronchial asthma phenotypes in children

The present paper analyses genetic predictors of various endotypes and phenotypes of bronchial asthma in children. The review of data on the structure and prevalence of single-nucleotide polymorphisms of interleukin genes demonstrates their correlation with the risk of virus-induced and allergen-induced phenotypes of bronchial asthma. Significant differences in genotypes correlate with aberrant production of interleukin and the risks for developing various phenotypes of the disease. The studies into genetic factors indicate the significance of functional polymorphisms of interleukin genes as predictors associated with phenotypes and risk of the disease.

DEPDC5 plays a vital role in epilepsy: Genotypic and phenotypic features in cohort and literature.

DEPDC5 emerges to play a vital role in focal epilepsy. However, genotype-phenotype correlation in DEPDC5-related focal epilepsies is challenging and controversial. In this study, we aim to investigate the genotypic and phenotypic features in DEPDC5-affected patients. Genetic testing combined with criteria published by the American College of Medical Genetics and Genomics and the Association for Molecular Pathology (ACMG/AMP), was used to identify pathogenic/likely pathogenic variants in DEPDC5 among the cohort of 479 patients with focal epilepsy. Besides, the literature review was performed to explore the genotype-phenotype correlation and the penetrance in DEPDC5-related focal epilepsies. Eight unrelated probands were revealed to carry different pathogenic/likely pathogenic variants in DEPDC5 and the total prevalence of DEPDC5-related focal epilepsy was 1.67% in the cohort. Sixty-five variants from 28 studies were included in our review. Combined with the cases reported, null variants accounted for a larger proportion than missense variants and were related to unfavorable prognosis (drug resistance or even sudden unexpected death in epilepsy; χ2 = 5.429, p = .020). And, the prognosis of probands with developmental delay/intellectual disability or focal cortical dysplasia was worse than that of probands with simple epilepsy (χ2 = -, p = .006). Besides, the overall penetrance of variants in DEPDC5 was 68.96% (231/335). The study expands the variant spectrum of DEPDC5 and proves that the DEPDC5 variant plays a significant role in focal epilepsy. Due to the characteristics of phenotypic heterogeneity and incomplete penetrance, genetic testing is necessary despite no specific family history. And we propose to adopt the ACMG/AMP criteria refined by ClinGen Sequence Variant Interpretation Working Group, for consistency in usage and transparency in classification rationale. Moreover, we reveal an important message to clinicians that the prognosis of DEPDC5-affected patients is related to the variant type and complications.

Virulent and Multi-drug-Resistant Edwardsiella tarda Infection in Oscar Fish: Unveiling the Threat of Mass Mortality and AMR Dissemination.

The Oscar fish (Astronotus ocellatus) is among the most commonly domesticated and exported ornamental fish species from Kerala. The ornamental fish industry faces a significant challenge with the emergence of diseases caused by multi-drug-resistant bacteria. In the present study, six isolates were resolved from the diseased Oscar fish showing haemorrhages, necrosis, and loss of pigmentation. After phenotypic and genotypic characterization, the bacteria were identified as Edwardsiella tarda, Klebsiella pneumoniae, Enterococcus faecalis, Escherichia coli, Brevibacillus borstelensis, and Staphylococcus hominis. Experimental challenge studies in healthy Oscar fish showed that E. tarda caused 100% mortality within 240h with 6.99 × 106CFU/fish as LD50 and histopathology revealed the typical signs of infection. The pathogen was re-recovered from the moribund fish thereby confirming Koch's postulates. E. tarda was confirmed through the positive amplification of tarda-specific gene and virulence genes viz., etfD and escB were also detected using PCR. Antibiotic susceptibility tests using disc diffusion displayed that the pathogen is multi-drug-resistant towards antibiotics belonging to aminoglycosides, tetracyclines, and quinolones categories with a MAR index of 0.32, which implicated the antibiotic pressure in the farm. Plasmid curing studies showed a paradigm shift in the resistance pattern with MAR index of 0.04, highlighting the resistance genes are plasmid-borne except for the chromosome-borne tetracycline resistance gene (tetA). This study is the first of its kind in detecting mass mortality caused by E. tarda in Oscar fish. Vigilant surveillance and strategic actions are crucial for the precise detection of pathogens and AMR in aquaculture.

Unlocking the Potential of Pepper Plants under Salt Stress: Mycorrhizal Effects on Physiological Parameters Related to Plant Growth and Gas Exchange across Tolerant and Sensitive Genotypes.

Agriculture is confronted with the challenge of ensuring global food security, yet the rapid expansion of salinity stress undoubtedly restricts plant productivity in cultivable areas, posing a significant threat to crop yields. Arbuscular mycorrhizal fungi (AMFs) have emerged as a biological tool for enhancing plant salt stress tolerance. To utilize this biological tool, this study evaluated the response in growth and physiological parameters of tolerant (Karaisali) and sensitive (Demre) pepper genotypes. The experiment involved mycorrhizal-treated (Glomus clarium) and non-mycorrhizal (control) plants of both the tolerant and sensitive pepper genotypes. The plants were subjected to two salt doses: 75 and 150 mM. The plant growth and physiological parameters were measured at 40 days after transplanting. The mycorrhizal activity was found to be significantly more effective in the sensitive genotype. We found notable differences in mycorrhizal activity between the pepper genotypes under salt stress conditions, with the sensitive genotype "Demre" showing greater responsiveness to mycorrhizal association compared with the "Karaisali" variety. Under both moderate (75 mM NaCl) and higher salt stress levels (150 mM NaCl), both the "Karaisali" and "Demre" varieties exhibited substantial increases in their shoot dry weights. However, these increases were consistently higher in the "Demre" plants. Moreover, the AMFs demonstrated significant enhancements in photosynthesis rates under both moderate and high salinity levels in both genotypes. Overall, our findings suggest that AMFs play a crucial role in improving plant growth, water status, and photosynthesis characteristics, particularly in salt-sensitive pepper genotypes, under moderate-to-high salinity levels. In conclusion, the plant growth, water status, and photosynthesis characteristics of the salt-sensitive pepper genotype were significantly improved by AMFs at medium and high salinity levels, such as 75 mM and 150 mM NaCl, respectively.

A New Mouse Model for Usher Syndrome Crossing Kunming Mice with CBA/J Mice

BackgroundPreviously, we discovered a strain of Kunming mice, referred to as the KMush/ush strain, that exhibited notably abnormal electroretinogram (ERG) readings and elevated thresholds for auditory brainstem responses (ABRs), which resembled the characteristics of Usher Syndrome (USH). We successfully identified the pathogenic genes, Pde6b and Adgrv1, after KMush/ush crossbred with CBA/CaJ mice, referred to as CBA-1ush/ush, CBA-2ush/ush or CBA-2ush/ush. In this investigation, we crossbred KMush/ush and CBA/J mice to establish novel recombinant inbred lines and analysed their phenotypic and genotypic characteristics. MethodsERG readings, ABR testing, fundus morphology, histological examination of the retina and inner ear, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis, western blotting, DNA sequence analysis and behavioural experiments were performed to assess the phenotypes and genotypes of the progeny lines. ResultsNo obvious waveforms in the ERG were detected in F1 hybrid mice while normal ABR results were recorded. The F2 hybrids, which were called J1ush/ush or J2ush/ush, exhibited segregated hearing-loss phenotypes. J1ush/ush mice had a retinitis pigmentosa (RP) phenotype with elevated ABR thresholds, whereas J2ush/ush mice exhibited only the RP phenotype. Interestingly, J1ush/ush mice showed significantly higher ABR thresholds than wild-type mice at 28 days post born (P28), and RT-qPCR and DNA-sequencing analysis showed that Adgrv1 gene expression was significantly altered in J1ush/ush mice, but histological analysis showed no significant structural changes in the organ of Corti or spiral ganglia. Further elevation of ABR-related hearing thresholds by P56 manifested only as a reduced density of spiral ganglion cells, which differed significantly from the previous pattern of cochlear alterations in CBA-2ush/ush mice. ConclusionsWe successfully introduced the hearing-loss phenotype of inbred mice with USH into CBA/J mice, which provides a good animal model for future studies on the important physiological roles of the Adgrv1 gene in inner-ear structure and for therapeutic studies targeting Adgrv1-mutated USH.

Chylomicron retention disease caused by SAR1B gene variations in 2 cases and literatures review

Objective: To summarize the genotype and clinical characteristics of chylomicron retention disease (CMRD) caused by secretion associated Ras related GTPase 1B (SAR1B) gene variations. Methods: Clinical data and genetic testing results of 2 children with CMRD treated at Children's Hospital of Fudan University and Jiangxi Provincial Children's Hospital from May 2022 to July 2023 were summarized. To provide an overview of the clinical and genetic characteristics of CMRD caused by SAR1B gene variations, all of the literature was searched and reviewed from China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, China VIP database, China Biology Medicine disc and PubMed database (up to January 2024) with "chylomicron retention disease" "Anderson disease" or "Anderson syndrome" as the search terms. All relevant literatures were reviewed to summarize the clinical and genetic features of CMRD caused by SAR1B gene variations. Results: One 11-year-old boy and one 4-month-old girl with CMRD. Both patients had lipid malabsorption, failure to thrive, decreased cholesterol, elevated transaminase and creatine kinase, and vitamin E deficiency, with homozygous variations (c.224A>G) and compound heterozygous variations (c.224A>G and c.554G>T) in SAR1B gene, respectively. Case 1 was followed up for over a month, and he still occasionally experienced lower limb muscle pain. Case 2 was followed up for more than a year, and her had caught up to normal levels. Both patients had no other significant discomfort. Literature search retrieved 0 Chinese literature and 22 English literatures. In addition to the 2 cases reported in this study, a total of 51 patients were identified as CMRD caused by SAR1B gene variations. Twenty-one types of SAR1B variants 10 missense, 4 nonsense, 3 frameshift, 1 in-frame deletion, 1 splice, 1 gross deletion, and 1 gross insertion-deletion were found among the 51 CMRD cases. Among all the patients, 49 cases had lipid malabsorption (43 cases had diarrhea or fatty diarrhea, 17 cases had vomiting, and 12 cases had abdominal distension), 45 cases had lipid soluble Vitamin deficiency (43 cases had Vitamin E deficiency, 10 cases had Vitamin A deficiency, 9 case had Vitamin D deficiency, and 5 cases had Vitamin K deficiency), 35 cases had failure to thrive, 32 cases had liver involvement (32 cases had elevated transaminases, 5 cases had fatty liver, and 3 cases had hepatomegaly), 29 cases had white small intestinal mucosa under endoscopy, and 17 cases had elevated creatine kinase, 14 cases had neuropathy, 5 cases had ocular lesions, 2 cases had acanthocytosis, 1 case had decreased cardiac ejection fraction, and 1 case was symptom-free. Conclusions: Early infancy failure to thrive and lipid malabsorption are common issues for CMRD patients. The Laboratory tests are characterized by hypocholesterolemia with or without fat-soluble vitamin deficiency, elevated liver enzymes and (or) creatine kinase. Currently, missense variations are frequent among the primarily homozygous SAR1B genotypes that have been described.

Phenotypic and Genotypic Characterization of Staphylococcus aureus Isolated from Nasal Samples of Healthy Dairy Goats in Algeria.

The present study aimed to determine the phenotypic and genotypic characteristics of S. aureus isolates from the nasal swabs of goats. A total of 232 nasal samples (one per animal) were collected from goats on 13 farms located in two regions of Algeria and were analyzed for the presence of S. aureus. The detection of virulence factors was carried out using PCR. The antibiotic susceptibility of the recovered isolates was assessed using the disc diffusion method. The biofilm formation ability was assessed by the Congo red agar method and a microtiter plate assay, and the molecular characterization of isolates was carried out by spa-typing, and for selected isolates also by multilocus sequence typing (MLST). Overall, 36 out of 232 nasal swabs (15.5%) contained S. aureus, and 62 isolates were recovered. Regarding the virulence factors, at least one staphylococcal enterotoxin gene was detected in 30 (48.4%) isolates. The gene tst encoding the toxic shock syndrome toxin was detected in fifteen isolates (24.2%), but none of the isolates harbored the gene of Panton-Valentine leukocidin (lukF/S-PV). Nine different spa-types were identified, including the detection of a new one (t21230). The recovered isolates were assigned to three clonal complexes, with CC5 (51.8%) being the most common lineage. Two isolates were methicillin-resistant (MRSA) and belonged to ST5 (CC5) and to spa-types t450 and t688. Moreover, 27 (43.5%) of the S. aureus isolates were found to be slime producers in Congo red agar, and all of the recovered isolates could produce biofilms in the microtiter plate assay. Our study showed that the nares of healthy goats could be a reservoir of toxigenic and antibiotic-resistant strains of S. aureus isolates, including MRSA, which could have implications for public health.

ОЦЕНКА СТАБИЛЬНОСТИ УРОЖАЙНОСТИ ЯРОВОГО ЯЧМЕНЯ

The purpose of research is to determine the share of influence of factors in the overall variability of barley yield and assess stability using parametric and non-parametric statistical methods in the conditions of the south of the Moscow Region. According to the results of the study (2020–2022), significant contributions to yield variability were identified: environmental conditions – 81.9 %; genotypic characteristics – 7.4 and genotype × environment interaction – 8.2 %. 7 samples (280.7–345.2 g/m2) were classified as excee¬ding the standard. Evaluation by parametric (bi – regression coefficient; S2dᵢ – deviation from regression; θi – average variance component; θ(i) – GE variance component; Wi2 – environmental valence; σi2 – stability variance; CV – coefficient of variation) and nonparametric (S(1,2,3,6) – rank statistics; NP(i) – non-parametric stability statistics; KR – rank sum) stability indicators allowed us to identify differences in the response of the sample under study to environmental conditions. A high correlation was noted between S(3) and NP(4) (r = 0,90), S(1) and S(2) (r = 0,96), S(6) and NP(4) (r = 0,98), Wi2 and σi2, (r = 1,00), Wᵢ2 and θᵢ (r = 1,00), σ2ᵢ and θᵢ (r = 1,00), Wᵢ2 and θ(i) (r = –1,00), σ2ᵢ, and θ(i) (r = –1,00), θᵢ and θ(i) (r = –1,00). Relationship with productivity in such indicators as S(1), S(2), S(3), NP(1), Wᵢ2, σ2ᵢ, S2dᵢ, CV, θᵢ, θ(ᵢ) was weak, the greatest association was noted with bᵢ (r = 0,60), NP(3) (r = –0,67), NP(4) (r = –0,67), S(6) (r = –0,69), KR (r = –0,71), NP(2) (r = –0,78). Samples with a combination of stability and high yield were identified: Alei (k-31363, Russia), Maximus (k-31366, Russia), Austris (k-31368, Latvia), Povolzhsky Luch (k-31392, Russia).