Prevalence of multidrug-resistant hypervirulent Klebsiella pneumoniae without defined hypervirulent biomarkers in Anhui, China: a new dimension of hypervirulence.

Klebsiella pneumoniae is an opportunistic pathogen that mainly causes nosocomial infections and hospital-associated pneumonia in elderly and immunocompromised people. However, multidrug-resistant hypervirulent K. pneumoniae (MDR-hvKp) has emerged recently as a serious threat to global health that can infect both immunocompromised and healthy individuals. It is scientifically established that plasmid-mediated regulator of mucoid phenotype genes (rmpA and rmpA2) and other virulence factors (aerobactin and salmochelin) are mainly responsible for this phenotype. In this study, we collected 23 MDR-hvKp isolates and performed molecular typing, whole genome sequencing, comparative genomic analysis, and phenotypic experiments, including the Galleria mellonella infection model, to reveal its genetic and phenotypic features. Meanwhile, we discovered two MDR-hvKp isolates (22122315 and 22091569) that showed a wide range of hypervirulence and hypermucoviscosity without rmpA and rmpA2 and any virulence factors. In phenotypic experiments, isolate 22122315 showed the highest hypervirulence (infection model) with significant mucoviscosity, and conversely, isolate 22091569 exhibited the highest mucoviscosity (string test) with higher virulence compared to control. These two isolates carried carbapenemase (blaKPC - 2), ß-lactamase (blaOXA - 1, blaTEM - 1B), extended-spectrum ß-lactamase (ESBL) genes (blaCTX - M - 15, blaSHV - 106), outer membrane protein-coding genes (ompA), fimbriae encoding genes (ecpABCDER), and enterobactin coding genes (entAB, fepC). In addition, single nucleotide polymorphism analysis indicated that both isolates, 22122315 and 22091569, were found to have novel mutations in loci FEBNDAKP_03184 (c. 2084A > C, p. Asn695Thr), and EOFMAFIB_02276 (c. 1930C > A, p. Pro644Thr), respectively. Finally, NCBI blast analysis suggested these mutations are located in the wzc of the capsule polysaccharide (cps) region and are responsible for putative tyrosine kinase. This study would be a strong reference for enhancing the current understanding of identifying the MDR-hvKp isolates that lacked both mucoid regulators and virulence factors.

Epidemiological characteristics of multidrug-resistant Acinetobacter baumannii ST369 in Anhui, China.

IMPORTANCE: Acinetobacter baumannii is a major health threat due to its antibiotic resistance and ability to cause nosocomial infections. Epidemiological studies indicated that the majority of globally prevalent ST369 clones originated from China, indicating a significant impact on public health in the country. In this study, we conducted whole-genome sequencing, comparative genomics, and Galleria mellonella infection model on eight A. baumannii ST369 isolates collected from a provincial hospital in China to comprehensively understand the organism. We identified two mutations (G540A and G667D) on the wzc gene that can affect bacterial virulence and viscosity. We confirmed their impact on resistance and virulence. We also investigated the potential involvement of AB46_0125 and AB152_03903 proteins in virulence. This finding provides a theoretical reference for further research on A. baumannii ST369 clinical isolates with similar mutations.

Assessment of the hypoglycemic and anti-hemostasis effects of Paederia foetida (L.) in controlling diabetes and thrombophilia combining in vivo and computational analysis.

Paederia foetida is valued for its folk medicinal properties. This research aimed to assess the acute toxicity, hypoglycemic and anti-hemostasis properties of the methanolic extract of P. foetida leaves (PFLE). Acute toxicity of PFLE was performed on a mice model. Hypoglycemic and anti-hemostasis properties of PFLE were investigated on normal and streptozotocin-induced mice models. Deep learning, molecular docking, density functional theory, and molecular simulation techniques were employed to understand the underlying mechanisms through in silico study. Oral administration of PFLE at a dosage of 300 µg/kg body weight (BW) showed no signs of toxicity. Treatment with PFLE (300 µg/kg/BW) for 14 days resulted in a hypoglycemic condition and a 30.47% increase in body weight. Additionally, PFLE mixed with blood exhibited a 44.6% anti-hemostasis effect. Deep learning predicted the inhibitory concentration (pIC50, nM) of Cleomiscosins against SGLT2 and FXa to be 7.478 and 6.017, respectively. Molecular docking analysis revealed strong binding interactions of Cleomiscosins with crucial residues of the target proteins, exhibiting binding energies of -8.2 kcal/mol and -7.1 kcal/mol, respectively. ADME/Tox predictions indicated favorable pharmacokinetic properties of Cleomiscosins, and DFT calculations of frontier molecular orbitals analyzed the stability and reactivity of these compounds. Molecular simulation dynamics, principal component analysis and MM-PBSA calculation demonstrated the stable, compact, and rigid nature of the protein-ligand complexes. The methanolic PFLE exhibited significant hypoglycemic and anti-hemostasis properties. Cleomiscosin may have inhibitory properties for the development of novel drugs to manage diabetes and thrombophilia in the near future.

Molecular identification and biological control of Ralstonia solanacearum from wilt of papaya by natural compounds and Bacillus subtilis: An integrated experimental and computational study.

Ralstonia solanacearum is a harmful pathogen that causes severe wilt disease in several vegetables. In the present study, we identified R. solanacearum from wilt of papaya by 16S rRNA PCR amplification. Virulence ability of R. solanacearum was determined by amplification of approximately 1500 bp clear band of hrpB gene. Further, in-vitro seed germination assay showed that R. solanacearum reduced the germination rate up to 26.21%, 34% and 33.63% of cucumber, bottle guard and pumpkin seeds, respectively whereas shoot and root growth were also significantly decreased. Moreover, growth inhibition of R. solanacearum was recorded using antibacterial compound from medicinal plant and antagonistic B. subtilis. Petroleum ether root extract of Rauvolfia serpentina showed highest 22 ± 0.04 mm diameter of zone of inhibition where methanolic extract of Cymbopogon citratus and ethanolic extract of Lantana camara exhibited 20 ± 0.06 mm and 20 ± 0.01 mm zone of inhibition against R. solanacearum, respectively. In addition, bioactive compounds of B. subtilis inhibited R. solanacearum growth by generating 17 ± 0.09 mm zone of inhibition. To unveil the inhibition mechanism, we adopted chemical-protein interaction network and molecular docking approaches where we found that, rutin from C. citratus interacts with citrate (Si)-synthase and dihydrolipoyl dehydrogenase of R. solanacearum with binding affinity of -9.7 kcal/mol and -9.5 kcal/mol while quercetin from B. subtillis interacts with the essential protein F0F1 ATP synthase subunit alpha of the R. solancearum with binding affinity of -6.9 kcal/mol and inhibit the growth of R. solanacearum. Our study will give shed light on the development of eco-friendly biological control of wilt disease of papaya.

Clinical manifestations and socio-demographic status of COVID-19 patients during the second-wave of pandemic: A Bangladeshi experience.

BACKGROUND: Bangladesh is a densely populated country with a substandard healthcare system and a mediocre economic framework. Due to the enormous number of people who have been unaware until now, the development of COVID-19's second-wave infection has become a severe threat. The present investigation aimed to characterize the clinical and socio-demographic characteristics of COVID-19 in Bangladesh. METHODS: A cross-sectional analysis was carried out from all the other COVID-19 patients and confirmed by RT-PCR undergoing a specialized COVID-19 hospital. From March 1 to April 15, 2021, a total of 1326 samples were collected. Samples were only obtained from non-critical COVID-19 patients as critically ill patients required emergency intensive care medications. Then, from April 17 to May 03, 2021, SARS-CoV-2 infection and clinical assessment was performed based on interim guidelines from the WHO. The diagnosis was conducted through RT-PCR. Later, identifying the symptomatic and asymptomatic patient based on checking the Clinical Observation Form (COF). The patients filled the COF form. Finally, statistical analyses were done using the SPSS 20 statistical program. RESULTS: In this investigation, a total of 326 patients were diagnosed as COVID-19 positive. Among them, approximately 19.02% (n = 62) were asymptomatic, and 80.98% (n = 264) were symptomatic. Here, the finding shows that the occurrence of this infection was varied depending on age, sex, residence, occupation, smoking habit, comorbidities, etc. However, Males (60.12%) were more affected than females (39.88%), and, surprisingly, this pandemic infected both urban and rural residents almost equally (urban = 50.92%; rural = 49.08%). Approximately 19% of the asymptomatic and 62% of symptomatic cases had at least one comorbid disorder. Interestingly, an unexpected result was exhibited in the case of smokers, where non-smokers were more affected than smokers. The study indicates community transmission of COVID 19, where people were highly infected at their occupations (35.58%), at houses (23.93%) and by traveling (12.88%). Noteworthy, according to this report, a large number (19.33%) of individuals did not know exactly how they were contaminated with SARS-CoV-2. Patients were most commonly treated by an antibiotic 95.09%, followed in second by corticosteroid 46.01%. Anti-viral drugs, remdesivir, and oxygenation are also needed for other patients. Among those, who were being treated, approximately 69.33% were isolated at home, 27.91% were being treated at dedicated COVID-19 hospitals. Finally, 96.63% were discharged without complications, and 0.03% has died. CONCLUSION: This investigation concludes that males became more infected than females. Interestingly, both urban and rural people became nearly equally infected. It noticed community transmission of SARS-CoV-2, where people were highly infected at their workplaces. A higher rate of silent transmission indicates that more caution is needed to identify asymptomatic patients. Most of the infected people were isolated at home whereas nearly one-fourth were treated at hospitals. Clinically, antibiotics were the most widely used treatment. However, the majority of the patients were discharged without complications. The current investigation would be helpful to understand the clinical manifestations and socio-demographic situations during the second wave of the COVID-19 pandemic in Bangladesh.

SARS-CoV-2 molecular identification and clinical data analysis of associated risk factors from a COVID-19 testing laboratory of a coastal region in Bangladesh.

BACKGROUND AND AIM: Outbreak of COVID-19 seems to have exacerbated across the globe, including Bangladesh. Scientific literature on the clinical data record of COVID-19 patients in Bangladesh is inadequate. Our study analyzes the clinical data of COVID-19 positive patients based on molecular identification and risk factor correlated with three variables (age, sex, residence) and COVID-19 prevalence in the four districts of Chattogram Division (Noakhali, Feni, Lakshmipur and Chandpur) with an aim to understand the trajectory of this pandemic in Chattogram, Southern Bangladesh. METHODS: A cross-sectional study is conducted in the context of RT-PCR-based COVID-19 positive 5,589 individuals diagnosed with SARS-CoV-2 infection from the COVID-19 testing laboratory, Abdul Malek Ukil Medical College, Noakhali-3800, Bangladesh. For molecular confirmation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), standard diagnostic protocols through real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) were conducted. Different patient demographics were analyzed using SPSS version 22 for exploring the relationship of three factors - age, sex, and residence with a cumulative number of COVID-19 positive cases and prevalence of COVID-19 in four districts in Chattogram division. The data was recorded between May to July, 2020. RESULTS: Among the three parameters, the present study revealed that 20-40 cohort had the highest incidence of infection rate (51.80%, n = 2895) among the different age groups. Among the infected individuals, 56.8% (n = 3177) were male and 43.2% (n = 2412) were female, denoting males being the most susceptible to this disease. Urban residents (52.7%, n = 2948) were more vulnerable to SARS-CoV-2 infection than those residing in rural areas (47.3%, n = 2641). The prevalence of COVID-19 positive cases among the four districts was recorded highest in the Noakhali district with 36.8% (n = 2057), followed by the Feni, Lakshmipur and Chandpur districts with 25.9% (n = 1448), 20.8% (n = 1163) and 16.5% (n = 921), respectively. CONCLUSIONS: This study presents a statistical correlation of certain factors linked to Bangladesh with confirmed COVID-19 patients, which will enable health practitioners and policy makers to take proactive steps to control and mitigate disease transmission.