Experimental evolution under different nutritional conditions changes the genomic architecture and virulence of Acinetobacter baumannii.

This study uncovers the molecular processes governing the adaptive evolution of multidrug-resistant (MDR) pathogens without antibiotic pressure. Genomic analysis of MDR Acinetobacter baumannii cells cultured for 8000 generations under starvation conditions (EAB1) or nutrient-rich conditions (EAB2) revealed significant genomic changes, primarily by insertion sequence (IS)-mediated insertions and deletions. Only two Acinetobacter-specific prophage-related deletions and translocations were observed in the EAB1 strain. Both evolved strains exhibited higher virulence in mouse infection studies, each with different modes of action. The EAB1 strain displayed a heightened ability to cross the epithelial barrier of human lung tissue, evade the immune system, and spread to lung tissues, ultimately resulting in cellular mortality. In contrast, the EAB2 strain strongly attached to epithelial cells, leading to increased synthesis of proinflammatory cytokines and chemokines. The genomic alterations and increased virulence observed in evolved strains during short-term evolution underscore the need for caution when handling these pathogens, as these risks persist even without antibiotic exposure.

Metabarcoding study of potential pathogens and zoonotic risks associated with dog feces in Seoul, South Korea.

BACKGROUND: A significant portion of South Korea's population, approximately a quarter, owns pets, with dogs being the most popular choice among them. However, studies analyzing the fecal organism communities of dogs in South Korea are lacking, and limited efforts have been exerted to identify pathogens with potential zoonotic implications. Therefore, this study aimed to investigate potential pathogens using metabarcoding analysis and evaluate the risk of zoonotic diseases in dog feces in Seoul, South Korea. METHODOLOGY: Fecal samples were collected from both pet and stray dogs in the Mapo district of Seoul. Next-generation sequencing (NGS) was utilized, employing 16S rRNA amplicon sequencing to identify prokaryotic pathogens, and 18S rRNA amplicon sequencing for eukaryotic pathogens. The data obtained from the QIIME2 pipeline were subjected to various statistical analyses to identify different putative pathogens and their compositions. PRINCIPAL FINDINGS: Significant variations in microbiota composition were found between stray and pet dogs, and putative prokaryotic and eukaryotic pathogens were identified. The most prevalent putative bacterial pathogens were Fusobacterium, Helicobacter, and Campylobacter. The most prevalent putative eukaryotic pathogens were Giardia, Pentatrichomonas, and Cystoisospora. Interestingly, Campylobacter, Giardia, and Pentatrichomonas were found to be significantly more prevalent in stray dogs than in pet dogs. The variation in the prevalence of potential pathogens in dog feces could be attributed to environmental factors, including dietary variances and interactions with wildlife, particularly in stray dogs. These factors likely contributed to the observed differences in pathogen occurrence between stray and pet dogs. CONCLUSIONS/SIGNIFICANCE: This study offers valuable insights into the zoonotic risks associated with dog populations residing in diverse environments. By identifying and characterizing putative pathogens in dog feces, this research provides essential information on the impact of habitat on dog-associated pathogens, highlighting the importance of public health planning and zoonotic risk management.

Microbiome of lovebug (Plecia longiforceps) in Seoul, South Korea.

Lovebugs appeared in large numbers across a wide area in Seoul, South Korea, in June 2023. The sudden appearance of exotic insects not only discomforts people but also fosters anxiety, as their potential for pathogen transmission would be unknown. In this study, targeted next-generation sequencing (NGS) of the 16S rRNA gene V4 region was performed using iSeq 100 to screen for bacteria in lovebugs. Forty-one lovebugs (20 females and 21 males) collected in Seoul, Korea, were identified as Plecia longiforceps based on mitochondrial cytochrome oxidase subunit 1 sequencing data using PCR. We analyzed the microbiome of the lovebugs and detected 453 species of bacteria. Among all bacteria screened based on NGS, Rickettsia was detected in all samples with an average relative abundance of 80.40%, followed by Pandoraea and Ewingella. Diversity (alpha and beta) between females and males did not differ; however, only Tumebacillus showed a higher relative abundance in females. Sequencing analysis of Rickettsia using a gltA gene-specific primer by PCR showed that it had higher sequence similarity to the Rickettsia symbiont of arthropods than to the spotted fever group rickettsiae. Eleven samples in which Pandoraea was detected by iSeq 100 were confirmed by PCR and exhibited 100% sequence identity to Pandoraea oxalativorans strain DSM 23570. Consequently, the likelihood of pathogen transmission to humans is low. The applied method may play a crucial role in swiftly identifying bacterial species in the event of future outbreaks of exotic insects that may be harmful to humans.IMPORTANCELovebugs have recently emerged in large numbers in Seoul, causing major concern regarding potential health risks. By performing the next-generation sequencing of the 16S rRNA gene V4 region, we comprehensively examined the microbiome of these insects. We identified the presence of numerous bacteria, including Rickettsia and Pandoraea. Reassuringly, subsequent tests confirmed that these detected bacteria were not pathogenic. The present study addresses health concerns related to lovebugs and shows the accuracy and efficiency of our detection technique. Such methods prove invaluable for rapidly identifying bacterial species during potential outbreaks of unfamiliar insects, thereby ensuring public safety.

Metabarcoding of pathogenic parasites based on copro-DNA analysis of wild animals in South Korea.

Four species of dominant wild animals, namely, Prionailurus bengalensis euptilurus, Nyctereutes procyonoides koreensis, Hydropotes inermis argyropus, and Sus scrofa coreanus, are hosts of potential infectious agents, including helminths and protozoa. Therefore, it is necessary to analyze the infectious agents present in these wild animals to monitor and control the spread of pathogens. In the present study, fecal samples from 51 wild animals were collected from the mountains of Yangpyeong, Hoengseong, and Cheongyang in South Korea and metabarcoding of the V9 region of the 18S rRNA gene was performed to identify various parasite species that infect these wild animals. Genes from nematodes, such as Metastrongylus sp., Strongyloides spp., Ancylostoma sp., and Toxocara sp., were detected in the fecal samples from wild animals. In addition, platyhelminthes, including Spirometra sp., Echinostomatidae gen. sp., Alaria sp., Neodiplostomum sp., and Clonorchis sp., and protozoa, including Entamoeba sp., Blastocystis sp., Isospora sp., Tritrichomonas sp., Pentatrichomonas sp., and Cryptosporidium sp., were detected. In the present study, various parasites infecting wild animals were successfully identified using metabarcoding. Our technique may play a crucial role in monitoring parasites within wild animals, especially those causing zoonoses.

Effectiveness of community-based interventions for older adults living alone: a systematic review and meta-analysis.

OBJECTIVES: This study examined the effectiveness of community-based interventions designed for older adults living alone through a systematic review and meta-analysis. METHODS: The study incorporated 4 randomized controlled trials (RCTs) and 5 non-RCTs to evaluate various interventions. The methodological quality of these studies was assessed using the Downs and Black checklist, while Q-statistic and I-square tests were performed to examine statistical heterogeneity. Additionally, visual inspection of funnel plots and the trim-and-fill method were employed to investigate potential publication bias. Of the 2,729 identified studies, 9 met the criteria for inclusion in this review. Independent variables were categorized into 5 groups (physical activity, nutrition, social relationships, social participation, and combined intervention) to examine their effects. Dependent variables were similarly classified into 5 subgroups to identify the specific effects of the interventions. RESULTS: Interventions focusing on nutrition and combined approaches were the most effective, yielding effect sizes of 0.96 (95% confidence interval [CI], 0.66 to 1.25) and 0.43 (95% CI, 0.26 to 0.60), respectively. The interventions had the greatest impacts on the health behavior and mental health of the participants, with effect sizes of 0.98 (95% CI, 0.73 to 1.22) for health behavior and 0.67 (95% CI, 0.19 to 1.16) for mental health. CONCLUSIONS: This study suggests a direction for the development of community-based interventions tailored to the needs of older adults living alone. Additionally, it provides evidence to inform policy decisions concerning this demographic.

The Mediating Role of Social Participation in Motoric Cognitive Risk and its Relation to Depression and Loneliness in Older Adults.

BACKGROUND: Motoric cognitive risk syndrome (MCR) reduces the quality of life, independence, and social interaction in older adults. Social participation is a potentially modifiable factor that benefits cognitive and mental health. This study explored the mediating roles of social participation between MCR and depression and between MCR and loneliness. METHODS: We performed a secondary analysis of data from the 2015-2016 National Social Life, Health, and Aging Project. Slow gait speed and cognitive decline were used to assess MCR. Mediation analysis was applied to two models, both of which used MCR as an exposure and social participation as a mediator. The outcomes were depression and loneliness for each model, respectively. RESULTS: Among 1,697 older adults, 196 (11.6%) had MCR. The mediating role of social participation was statistically significant in both models. The indirect effect (ß=0.267, p=0.001) of MCR on depression through social participation comprised 11.97% of the total effect (ß=2.231, p<0.001). The indirect effect (ß=0.098, p=0.001) of MCR on loneliness through social participation was 19.48% of the total effect (ß=0.503, p<0.001). CONCLUSION: Interventions to increase social participation may reduce depression and loneliness of older adults with MCR.

Blue Light Sensing BlsA-Mediated Modulation of Meropenem Resistance and Biofilm Formation in Acinetobacter baumannii.

The presence or absence of BlsA, a protein with a blue light-sensing flavin domain in the genomes of Acinetobacter species has aroused curiosity about its roles in the regulation of bacterial lifestyle under light. Genomic and transcriptomic analyses revealed the loss of BlsA in several multidrug-resistant (MDR) A. baumannii strains as well as the light-mediated induction of blsA, along with a possible BlsA-interacting partner BipA. Their direct in vivo interactions were verified using a bacterial two-hybrid system. The results demonstrated that the C-terminal region of BipA could bind to the C-terminal residues of BlsA under blue light at 23°C but not at 37°C. Genetic manipulations of blsA and bipA revealed that the coexistence of BlsA and BipA was required to induce the light-dependent expression of ompA in A. baumannii ATCC 17978 at 23°C. The same phenomenon occurred in the BlsA-deficient MDR strain in our functional complementation assay; however, the underlying molecular mechanism remains poorly understood. BlsA-modulated amounts of OmpA, the most abundant porin, in the outer membrane affected the membrane integrity and permeability of small molecules. Dark conditions or the deletion of ompA made the membrane more permeable to lipophilic ethidium bromide (EtBr) but not to meropenem. Interestingly, light illumination and low temperature conditions made the cells more sensitive to meropenem; however, this bactericidal effect was not noted in the blsA mutant or in the BlsA-deficient MDR strains. Light-mediated cell death and the reduction of biofilm formation at 23°C were abolished in the blsA mutant strain, suggesting multifaceted roles of BlsA in A. baumannii strains. IMPORTANCE Little is known about the functional roles of BlsA and its interacting partners in Acinetobacter species. Intriguingly, no BlsA homolog was found in several clinical isolates, suggesting that BlsA was not required inside the host because of the lack of blue light and the warm temperature conditions. As many chromophore-harboring proteins interact with various partners to control light-dependent cellular behaviors, the maintenance of blsA in the genomes of many Acinetobacter species during their evolution may be beneficial when fluctuations occur in two important environmental factors: light and temperature. Our study is the first to report the novel protein partner of BlsA, namely, BipA, and its contribution to multiple phenotypic changes, including meropenem resistance and biofilm formation. Rapid physiological acclimation to changing light or temperature conditions may be possible in the presence of the light-sensing BlsA protein, which may have more interacting partners than expected.

Zoo animal manure as an overlooked reservoir of antibiotic resistance genes and multidrug-resistant bacteria.

Animal fecal samples collected in the summer and winter from 11 herbivorous animals, including sable antelope (SA), long-tailed goral (LTG), and common eland (CE), at a public zoo were examined for the presence of antibiotic resistance genes (ARGs). Seven antibiotics, including meropenem and azithromycin, were used to isolate culturable multidrug-resistant (MDR) strains. The manures from three animals (SA, LTG, and CE) contained 104-fold higher culturable MDR bacteria, including Chryseobacterium, Sphingobacterium, and Stenotrophomonas species, while fewer MDR bacteria were isolated from manure from water buffalo, rhinoceros, and elephant against all tested antibiotics. Three MDR bacteria-rich samples along with composite samples were further analyzed using nanopore-based technology. ARGs including lnu(C), tet(Q), and mef(A) were common and often associated with transposons in all tested samples, suggesting that transposons carrying ARGs may play an important role for the dissemination of ARGs in our tested animals. Although several copies of ARGs such as aph(3')-IIc, blaL1, blaIND-3, and tet(42) were found in the sequenced genomes of the nine MDR bacteria, the numbers and types of ARGs appeared to be less than expected in zoo animal manure, suggesting that MDR bacteria in the gut of the tested animals had intrinsic resistant phenotypes in the absence of ARGs.

Constitutive Phenotypic Modification of Lipid A in Clinical Acinetobacter baumannii Isolates.

The degree of polymyxin B (PMB) resistance was measured in 40 clinical Acinetobacter baumannii isolates obtained from health care facilities. All of the tested isolates possessed a multidrug-resistant (MDR) phenotype against four classes of antibiotics (meropenem, doxycycline, gentamicin, and erythromycin), except for PMB. The blaOXA-23 gene was detected throughout the genetic analysis and experimental assay, indicating that all of the MDR strains were carbapenem-resistant A. baumannii strains. Multilocus sequence typing-based genotyping revealed that nine selected strains belonged to the international clone II lineage. When matrix-assisted laser desorption ionization-time of flight mass spectrometry was performed, intrinsic lipid A modification by phosphoethanolamine (PEtN) incorporation was noticeable only in the PMB-resistant (PMBR) strains. However, the presence of hexa- and penta-acylated lipid A due to the loss of the laurate (C12) acyl chain was noted in all PMB-susceptible strains but not in the PMBR strains. The reduction of negative surface charges in the PMBR strains was assessed by zeta potential analysis. Fluorescence imaging using dansyl-PMB revealed that, in the PMBR strains, PMB was less likely to bind to the cell surface. IMPORTANCE The widespread presence of MDR pathogens, including A. baumannii, is causing serious hospital-acquired infections worldwide. Extensive surveillance of MDR clinical A. baumannii isolates has been conducted, but the underlying mechanisms for their development of MDR phenotypes are often neglected. Either lipid A modification or loss of lipopolysaccharide in Gram-negative bacteria leads to PMBR phenotypes. The prevalence of intrinsic lipid A modification in PMBR clinical strains was attributed to high levels of basal expression of pmrC and eptA-1. Our findings suggest that new therapeutic strategies are warranted to combat MDR pathogens due to the emergence of many PMBR clinical strains.