Effect of Loulianwan on Gut Microbiota in db/db Mice with Type 2 Diabetes Mellitus Based on 16S rRNA Sequencing Technology / 中国实验方剂学杂志

ObjectiveTo investigate the effect of Loulianwan on the gut microbiota of db/db mice with type 2 diabetes mellitus （T2DM）. MethodMale db/m+ mice aged 4-5 weeks were assigned to the normal group， and male db/db model mice of the same age were randomly divided into model group， metformin group （0.25 g·kg-1·d-1）， and Loulianwan group （13 g·kg-1·d-1）， with six mice in each group. Drug intervention lasted five weeks. The body weight， water intake， and fasting blood glucose （FBG） of the mice were recorded every week. After five weeks， the FBG， liver triglyceride （TG）， liver total cholesterol （TC）， glycated serum protein （GSP）， and fasting serum insulin （FINS） were detected， and the insulin resistance index （HOMA-IR） was calculated. The feces in the mouse intestines were collected， and the 16S rRNA sequencing technology was used to detect the structural changes in the fecal gut microbiota of mice in each group. ResultCompared with the normal group， the model group showed increased body weight， water intake， FBG， liver TG， liver TC， GSP， FINS， and HOMA-IR （P<0.01）. Compared with the model group， the Loulianwan group showed reduced water intake， FBG， liver TG， liver TC， GSP， FINS， and HOMA-IR （P<0.01）. The gut microbiota in the Loulian Lills group changed from phylum to genus level. The relative abundance of beneficial bacteria increased and the relative abundance of harmful bacteria decreased. Among them， the abundance of Akkermansia muciniphila， Blautia， Ruminococcus， and Parabacteroides increased （P<0.01）. ConclusionLoulianwan can significantly improve glucose and lipid metabolism in db/db mice with T2DM， and its mechanism may be related to the increase in the abundance of Akkermansia muciniphila， Blautia， Ruminococcus， and Parabacteroides in the intestine.

Effects of Shaoyao gancao decoction on intestinal flora in rats with acute lung injury / 中国药房

OBJECTIVE To study the improvement effects of Shaoyao gancao decoction （SGD） on acute lung injury （ALI） in rats and its effects on the intestinal flora. METHODS Sixty SD rats were randomly divided into normal group （CON group， normal saline）， model group （MOD group， normal saline）， positive control group （DEX group， 5 mg/kg dexamethasone）， SGD low-dose， medium-dose and high-dose groups （SGD-L， SGD-M， SGD-H groups， 5.8， 11.6， 23.2 g/kg， calculated by crude drug）， with 10 rats in each group. Each group was given relevant medicine 10 mL/kg intragastrically， for 7 consecutive days. Thirty minutes after the last administration， CON group was given constant volume of normal saline via airway infusion， and other groups were given lipopolysaccharide （5 mg/kg） via airway infusion to induce ALI model. After 12 hours of modeling， the lung tissue wet/dry weight ratio was calculated， and the contents of interleukin 1β （IL-1β）， IL-6 and tumor necrosis factor α（TNF-α） in rat bronchial alveolar lavage fluid （BALF） were all detected； the pathological changes of lung tissue were observed after hematoxylin-eosin staining. The intestinal flora of rat feces was analyzed by 16S rRNA sequencing technology， and the correlation of differential bacteria genera with inflammatory factors was also analyzed. RESULTS Compared with MOD group， the infiltration of inflammatory cells in the lung tissue of rats in each SGD dose group was decreased， and the thickening of alveolar septum and pulmonary edema improved； lung tissue wet/dry weight ratio， the levels of IL-1β， IL-6 and TNF-α in BALF significantly decreased （P＜0.05 or P＜0.01）. SGD （low dose） could improve the intestinal flora disorder in ALI rats， restore the diversity and richness of intestinal flora， regulate the structure of flora， reduce the abundance of Lactobacillus， Streptococcus and Escherichia-Shigella， and increase the abundance of Firmicutes， Lachnospira， Ruminococcus， Clostridia，Dubosiella and Akkermansia. Through correlation analysis， it was found that the relative abundance of Lactobacillus， Streptococcus and Escherichia-Shigella was positively related to the levels of inflammatory factor IL-1β， IL-6 and TNF-α （P＜0.05 or P＜0.01）. The relative abundance of Lachnospira， Dubosiella， Firmicutes was significantly negatively correlated with the levels of inflammatory factors mentioned above （P＜0.05 or P＜0.01）. CONCLUSIONS SGD may improve ALI by reducing lung tissue injury and inflammatory response and regulating flora structure in rats.

Effect of Danggui Niantongtang on Intestinal Flora in Adjuvant-induced Arthritis Rats of Wind-dampness-heat Arthralgia：Based on 16S rDNA Sequencing / 中国实验方剂学杂志

ObjectiveTo explore the mechanism of Danggui Niantongtang （DGNTT） against adjuvant-induced arthritis （AA） in rats with wind-dampness-heat arthralgia （FSR） based on the variation of intestinal flora. MethodA total of 60 SD rats were randomized into normal （control） group， FSR group， low-， medium-， and high-dose DGNTT （5.67， 11.34， 22.68 g·kg-1） groups， and methotrexate （MTX） group （1.35 mg·kg-1）， with 10 rats in each group. The rats， except the control group， were injected with Mtb adjuvant and then exposed to artificial climatic chamber （hot and humid with wind） for 64 h for modeling. The rats were treated with water， DGNTT or MTX for 28 days from the day of injection. Arthritis index （AI） of rats was measured and paw volume was determined with a volume meter. The morphology of synovial tissues of the knees was observed based on hematoxylin-eosin （HE） staining and the changes of intestinal flora were analyzed based on 16S rRNA sequencing. ResultDGNTT can alleviate the hyperplasia of synovial tissue and inflammation of AA rats with FSR and inhibit the formation of pannus. The results of 16S rRNA sequencing showed that the relative abundance of Firmicutes， Lactobacillus， Prevotella 9， and Alloprevotella decreased （P<0.05， P<0.01） and the relative abundance of Bacteroidetes and Bacteroides increased （P<0.01） in FSR group compared those in the control group. Compared with the FSR group， all DGNTT groups and MTX group had high relative abundance of Lactobacillus （P<0.05， P<0.01） and low relative abundance of Bacteroidetes （P<0.01） and medium-dose and high-dose DGNTT groups and MTX group showed high abundance of Firmicutes， Prevotella 9， and Alloprevotella and low abundance of Bacteroides （P<0.05， P<0.01）. Spearman's correlation analysis suggested that the abundance of Bacteroides and Helicobacter was in positive correlation with AI （P<0.05）， while the abundance of Prevotella 9 and Candidatus Saccharimonas was in negative correlation with AI （P<0.01， P<0.05）. There was a negative correlation between the abundance of Prevotella 9 and paw volume （P<0.01）， and the abundance of Ruminococcaceae NK4A214 group， Christensenellaceae R-7 group， and Bacteroides was in negative correlation with spleen index （P<0.05）. The abundance of Prevotella 9 was in negative correlation with spleen index （P<0.01）. ConclusionDGNTT is effective for arthritis with FSR， as it can regulate the composition of intestinal flora in AA rats by increasing the abundance of probiotics and inhibiting the growth of pathogenic bacteria. The mechanism is the likelihood that it improves intestinal immune metabolism to ensure intestinal homeostasis.

Intestinal and pharyngeal microbiota in early neonates: an analysis based on high-throughput sequencing / 中国当代儿科杂志

OBJECTIVES@#To investigate the distribution characteristics and correlation of intestinal and pharyngeal microbiota in early neonates.@*METHODS@#Full-term healthy neonates who were born in Shanghai Pudong New Area Maternal and Child Health Hospital from September 2021 to January 2022 and were given mixed feeding were enrolled. The 16S rRNA sequencing technique was used to analyze the stool and pharyngeal swab samples collected on the day of birth and days 5-7 after birth, and the composition and function of intestinal and pharyngeal microbiota were analyzed and compared.@*RESULTS@#The diversity analysis showed that the diversity of pharyngeal microbiota was higher than that of intestinal microbiota in early neonates, but the difference was not statistically significant (P>0.05). On the day of birth, the relative abundance of Proteobacteria in the intestine was significantly higher than that in the pharynx (P<0.05). On days 5-7 after birth, the relative abundance of Actinobacteria and Proteobacteria in the intestine was significantly higher than that in the pharynx (P<0.05), and the relative abundance of Firmicutes in the intestine was significantly lower than that in the pharynx (P<0.05). At the genus level, there was no significant difference in the composition of dominant bacteria between the intestine and the pharynx on the day of birth (P>0.05), while on days 5-7 after birth, there were significant differences in the symbiotic bacteria of Streptococcus, Staphylococcus, Rothia, Bifidobacterium, and Escherichia-Shigella between the intestine and the pharynx (P<0.05). The analysis based on the database of Clusters of Orthologous Groups of proteins showed that pharyngeal microbiota was more concentrated on chromatin structure and dynamics and cytoskeleton, while intestinal microbiota was more abundant in RNA processing and modification, energy production and conversion, amino acid transport and metabolism, carbohydrate transport and metabolism, coenzyme transport and metabolism, and others (P<0.05). The Kyoto Encyclopedia of Genes and Genomes analysis showed that compared with pharyngeal microbiota, intestinal microbiota was more predictive of cell motility, cellular processes and signal transduction, endocrine system, excretory system, immune system, metabolic diseases, nervous system, and transcription parameters (P<0.05).@*CONCLUSIONS@#The composition and diversity of intestinal and pharyngeal microbiota of neonates are not significantly different at birth. The microbiota of these two ecological niches begin to differentiate and gradually exhibit distinct functions over time.

Assessing multiple factors affecting the gut microbiome structure of very preterm infants

The composition and diversity of the gut microbiota are essential for the health and development of the immune system of infants. However, there is limited information on factors that influence the gut microbiota of very preterm infants. In this study, we analyzed factors that affect the gut microbiota of very preterm infants. The stool samples from 64 very preterm infants with a gestational age less than 32 weeks were collected for 16S rRNA gene sequencing. The infants were divided according to the delivery mode, antibiotic use during pregnancy, and feeding methods. The abundance of Proteobacteria was high in both cesarean (92.7%) and spontaneous (55.5%) delivery groups and then shifted to Firmicutes after the first week of birth. In addition, Proteobacteria was also the dominant phylum of infant gut microbiome for mothers with antibiotic use, with more than 50% after the first week of birth. In comparison, the dominant phylum for mothers without antibiotic use was Firmicutes. Proteobacteria level was also high in breastfeeding and mixed-feeding groups, consisting of more than 90% of the community. By contrast, Proteobacteria was the dominant phylum at the first week of birth but then shifted to Firmicutes for the formula-fed group. The alterations of gut microbiota in infants can affect their health condition during growth. This study confirmed that the different feeding types, delivery modes, and use of antibiotics during pregnancy can significantly affect the composition of the gut microbiota of very preterm infants.

Structural Characterization of Vaginal Microbiota in Threatened Abortion Patients with Syndrome of Kidney Deficiency by 16S rRNA Sequencing / 中国实验方剂学杂志

ObjectiveTo investigate the structural features of vaginal microbiota in the early threatened abortion patients with the syndrome of kidney deficiency. MethodThirty-one patients with early threatened abortion of kidney deficiency syndrome （DK-TA group） and 116 women with normal early pregnancy （NP group） attending the First Affiliated Hospital of Guangzhou University of Chinese Medicine from May 2018 to December 2020 were selected. The vaginal secretions were collected for 16S rRNA sequencing， which can reveal the vaginal microbiota composition and differential bacteria between the two groups. ResultThe DK-TA group had higher abundance and diversity of vaginal microbiota than the NP group. The Binary jaccard and unweighted_unifrac distance matrix analysis showed that the similarity， dispersion， abundance， and phylogenetic relationship of vaginal microbiota were significantly different between the two groups. At the phylum level， the DK-TA group had lower relative abundance of Bacteroidetes and Fusobacteria and higher relative abundance of Proteobacteria and Gemmatimonadetes than the NP group. At the genus level， the DK-TA group had lower relative abundance of Sneathia and Bifidobacterium and higher relative abundance of Escherichia-Shigella and Shuttleworthia than the NP group. Linear discriminant analysis Effect Size（LEfSe）revealed that Bacteroidetes， Fusobacteria， and Bifidobacteria were dominant in the NP group and Proteobacteria and Firmicutes in the DK-TA group. The function prediction found that DK-TA was closely associated with 38 functional pathways， including cyclic adenosine monophosphate signaling pathway and regulation of tryptophan channels by inflammatory mediators. In addition， the vaginal differential bacteria between the two groups had significant positive or negative association with the differential metabolic pathways. ConclusionProteobacteria， Gemmatimonadetes， and Bacteroidetes in the vaginal microbiota may be biomarkers for threatened abortion of kidney deficiency syndrome.

Determining the protective effects of Ma-Mu-Ran Antidiarrheal Capsules against acute DSS-induced enteritis using 16S rRNA gene sequencing and fecal metabolomics / 中国天然药物

Ma-Mu-Ran Antidiarrheal Capsules (MMRAC) is traditional Chinese medicine that has been used to treat diarrhea caused by acute enteritis (AE) and bacillary dysentery in Xinjiang (China) for many years. However, the potential therapeutic mechanism of MMRAC for AE and its regulatory mechanism on host metabolism is unclear. This study used fecal metabolomics profiling with GC/MS and 16S rRNA gene sequencing analysis to explore the potential regulatory mechanisms of MMRAC on a dextran sulfate sodium salt (DSS)-induced mouse model of AE. Fecal metabolomics-based analyses were performed to detect the differentially expressed metabolites and metabolic pathways. The 16S rRNA gene sequencing analysis was used to assess the altered gut microbes at the genus level and for functional prediction. Moreover, Pearson correlation analysis was used to integrate differentially expressed metabolites and altered bacterial genera. The results revealed that six intestinal bacteria and seven metabolites mediated metabolic disorders (i.e., metabolism of amino acid, carbohydrate, cofactors and vitamins, and lipid) in AE mice. Besides, ten altered microbes mediated the differential expression of eight metabolites and regulated these metabolisms after MMRAC administration. Overall, these findings demonstrate that AE is associated with metabolic disorders and microbial dysbiosis. Further, we present that MMRAC exerts protective effects against AE by improving host metabolism through the intestinal flora.

Effects of nano titanium dioxide on gut microbiota based on human digestive tract microecology simulation system in vitro / 北京大学学报(医学版)

OBJECTIVE@#To explore the effects of oral exposure to titanium dioxide nanoparticles (TiO2 NPs) on the composition and structure of human gut microbiota.@*METHODS@#The particle size, shape, crystal shape and degree of agglomeration in ultrapure water of TiO2 NPs were characterized. The in vitro human digestive tract microecological simulation system was established by simulating the fluid environment and physical conditions of stomach, small intestine and colon, and the stability of the simulation system was evaluated. The bacterial communities were extracted from human feces and cultured stably in the simulated system. They were exposed to 0, 20, 100 and 500 mg/L TiO2 NPs, respectively, and the bacterial fluids were collected after 24 h of exposure. The effect of TiO2 NPs on the composition and structure of human gut microbiota was analyzed by 16S rRNA sequencing technology. Linear discriminant analysis effect size (LEfSe) was used to screen differential bacteria, and the Kyoto encyclopedia of genes and genomes (KEGG) database for functional prediction.@*RESULTS@#The spherical and anatase TiO2 NPs were (25.12±5.64) nm in particle size, while in ultra-pure water hydrated particle size was (609.43±60.35) nm and Zeta potential was (-8.33±0.22) mV. The in vitro digestive tract microecology simulation system reached a relatively stable state after 24 hours, and the counts of Enterococci, Enterobacte-rium, and Lactobacillus reached (1.6±0.85)×107, (5.6±0.82)×107 and (2.7±1.32)×107, respectively. 16S rRNA sequencing results showed that compared with the control group, the number and evenness of gut microbiota were not significantly affected at phylum, class, order, family and genus levels in TiO2 NPs groups (20, 100 and 500 mg/L). The relative abundance of some species was significantly changed, and a total of 42 different bacteria were screened between the TiO2 NPs groups (20, 100 and 500 mg/L) and the control group [linear discriminant analysis(LDA) score>3], represented by Enterobacter, Bacteroidaceae, Lactobacillaceae, Bifidobacteriaceae and Clostridium. Further predictive analysis of gut microbiota function showed that TiO2 NPs might affect oxidative phosphorylation, energy meta-bolism, phosphonate and phosphonate metabolism, and methane metabolism (P < 0.05).@*CONCLUSION@#In human digestive tract microecological simulation system, TiO2 NPs could significantly change the composition and structure of human gut microbiota, represented by Enterobacter and probiotics, and may further affect a variety of metabolism and function of the body.

Characteristics of gut microbiota in hyperuricemia population based on full-length 16S rRNA sequencing / 中华微生物学和免疫学杂志

Objective:To analyze the differences in gut microbiota between patients with hyperuricemia (HUA) and healthy population for better understanding the correlation between gut microflora and high uric acid.Methods:This study recruited 63 adult volunteers, including 25 HUA patients (HUA group) and 38 healthy people (control group), who underwent physical examination in the PLA Strategic Support Force Characteristic Medical Center in 2021. Their fecal samples were collected and analyzed by 16S rRNA high-throughput full-length gene sequencing to analyze the composition of gut microbiota.Results:The overall composition of gut microbiota was different between HUA group and control group. The α diversity index decreased significantly in HUA group and β diversity analysis showed that there were significant differences between the two groups. HUA group showed increased Bacteroidetes and decreased Firmicutes. LEfSe analysis indicated a unique microbiota structure in HUA group, characterized by significantly decreased short-chain fatty acid (SCFA)-producing bacteria represented by Faecalibacterium prausnitzii and significantly increased Streptococcus salivarius, Bacteroides fragilis, Fusobacterium hwasookii, Flavonifractor plautii, Mycobacterium mucogenicum B and Blautia sp003287895. In addition, functional prediction through PICRUSt2 showed that the metabolism related to gut microbiota SCFA pathway in HUA group was decreased, which was consistent to the unique microbiota structure. Conclusions:Compared with healthy population, patients with hyperuricemia had different composition of gut microbiota and metabolic feature.

The Relationship Between the Gut Microbiome and Neurodegenerative Diseases / 神经科学通报·英文版

Many recent studies have shown that the gut microbiome plays important roles in human physiology and pathology. Also, microbiome-based therapies have been used to improve health status and treat diseases. In addition, aging and neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease, have become topics of intense interest in biomedical research. Several researchers have explored the links between these topics to study the potential pathogenic or therapeutic effects of intestinal microbiota in disease. But the exact relationship between neurodegenerative diseases and gut microbiota remains unclear. As technology advances, new techniques for studying the microbiome will be developed and refined, and the relationship between diseases and gut microbiota will be revealed. This article summarizes the known interactions between the gut microbiome and neurodegenerative diseases, highlighting assay techniques for the gut microbiome, and we also discuss the potential therapeutic role of microbiome-based therapies in diseases.

Genital infection by Aerococcus viridans in a captive african elephant (Loxodonta africana) / Infecção genital por Aerococcus viridans em um elefante africano em cativeiro (Loxodonta africana)

ABSTRACT: Aerococcus viridans is an emerging pathogen for humans and livestock animals, mainly associated with genitourinary infections cases. Its occurrence in wild mammals has never been reported. The aim of this study was to determine the etiological agent associated with clinical a case of a genital infection in a female African elephant (Loxodonta africana). Phylogenetic analysis and antimicrobial susceptibility profile of the isolate were also addressed. The animal presented frequent cases of genital infection with intermittent white secretion. Purulent secretion was sampled and submitted to bacteriological exam. The isolate obtained was thus identified by phenotypic and molecular methods as A. viridans and was found to be similar to human pathogenic isolates in BLASTn and phylogenetic analysis. The isolate was sensitive to almost all antimicrobials evaluated, presenting resistance to ciprofloxacin and norfloxacin. This is the first report of occurrence of A. viridans infection in the genital tract of an African elephant.

RESUMO: Aerococcus viridans é um patógeno emergente para seres humanos e animais de produção, principalmente associado a casos de infecções geniturinárias. Sua ocorrência em mamíferos selvagens nunca foi relatada. O objetivo deste estudo foi determinar o agente etiológico associado a um caso clínico de infecção genital em uma fêmea de elefante africano (Loxodonta africana). Análises filogenéticas e perfil de susceptibilidade antimicrobiana do isolado também foram avaliados. O animal apresentou casos frequentes de infecção genital com eliminação de secreção branca intermitente. A secreção purulenta foi coletada e submetida a exame bacteriológico. O isolado obtido foi identificado por métodos fenotípicos e moleculares como A. viridans e apresentou alta similaridade a isolados humanos patogênicos nas análises de BLASTn e filogenética. O isolado foi sensível a quase todos os antimicrobianos avaliados, apresentando resistência à ciprofloxacina e norfloxacina. Este é o primeiro relato de ocorrência de infecção por A. viridans no trato genital de elefante africano.

The Relationship Between the Gut Microbiome and Neurodegenerative Diseases / 神经科学通报·英文版

Many recent studies have shown that the gut microbiome plays important roles in human physiology and pathology. Also, microbiome-based therapies have been used to improve health status and treat diseases. In addition, aging and neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease, have become topics of intense interest in biomedical research. Several researchers have explored the links between these topics to study the potential pathogenic or therapeutic effects of intestinal microbiota in disease. But the exact relationship between neurodegenerative diseases and gut microbiota remains unclear. As technology advances, new techniques for studying the microbiome will be developed and refined, and the relationship between diseases and gut microbiota will be revealed. This article summarizes the known interactions between the gut microbiome and neurodegenerative diseases, highlighting assay techniques for the gut microbiome, and we also discuss the potential therapeutic role of microbiome-based therapies in diseases.

Evaluation of the biocontrol potential of endophytic bacteria isolated from Coffea liberica (w. Bull ex hiern) against brown eyespot-causing fungal phytopathogen

Aims@#The study aims to investigate the potential of the endophytic bacteria as an alternative to control the devastating brown eyespot disease caused by Cercospora sp. in coffee plants. The fungal phytopathogen causes severe leaf fall and berry damages resulting in serious yield losses in coffee farms in the Philippines and worldwide. Currently, the management of this fungal infection relies heavily on synthetic fungicides, which may be of major environmental concern. @*Methodology and results@#Endophytic bacteria were isolated from the intercellular tissues of Coffea liberica leaves by surface sterilization, maceration, dilution technique, plating on trypticase soy agar and colony characterization. Fourteen isolated endophytic bacteria were screened for their ability to inhibit the mycelial growth of Cercospora sp. through modified dual culture assay. Isolates HCC10-3SC3, HCC10-3SC2, HCC10-1SC1, ICC10-3SC1, and ICC10-1SC1 yielded the highest percent inhibition of radial growth (PIRG) with 59.56%, 60.92%, 60.96%, 64.36%, and 67.06% respectively and are statistically significant (p<0.05) compared to the antibiotic control nystatin. The top five performing endophytic bacteria were subjected to hydrolytic enzyme production assays and found to exhibit amylolytic, lipolytic, proteolytic, chitinolytic, and cellulolytic activities. Based on the morphological and molecular identification by the 16S rRNA sequence analysis, isolates showed the similarity with Staphylococcus cohnii, Bacillus siamensis, Staphylococcus hominis, and Kosakonia cowanii found in GENBANK. @*Conclusion, significance and impact of study@#The study revealed the biological control potential of endophytic bacteria agents against the brown eyespot-causing fungus in coffee.

Characteristics of cervical microecology in late reproductive-age women with different grades of cervical lesions / 南方医科大学学报

OBJECTIVE@#To analyze the characteristics of cervical microecology in late reproductive-age women with cervical lesions and explore new methods for preventing cervical lesions.@*METHODS@#Cervical smears were obtained from a total of 147 women of late reproductive age, including 24 with high-risk HPV infection (HR-HPV), 27 with low-grade squamous intra-epithelial lesions (LSIL), 36 with high-grade squamous intra-epithelial lesions (HSIL), 35 with cervical cancer (CC) and 25 healthy women. llumina MiSeq sequencing of V3-V4 region of the 16S rRNA gene amplicons was used to characterize the vaginal microbiota of the women. OTUs analysis of the valid data was performed, and the α-diversity (Chao1, Simpson's Index and Shannon Index) and β-diversity (T-test, weighted UniFrac β diversity, and MetaStat analysis) were evaluated.@*RESULTS@#Dilution curve and species accumulation boxplot validated the quality of the samples. OTUs analysis of the 5 groups demonstrated that cervical bacterial genus consisted primarily of @*CONCLUSIONS@#The abundance of

A case report of Nocardia cyriacigeorgica presenting as cellulitis with multiple discharging sinuses

Nocardia spp. are filamentous Gram positive bacteria that are ubiquitous soil saprophytes. The majority of nocardial infections occur in severely immunocompromised patients who are particularly susceptible to pulmonary disease and dissemination. Extrapulmonary nocardiosis is relatively common and can occur through hematogenous dissemination or a contiguous spread of necrotizing pneumonitis. Primary cutaneous and soft tissue nocardiosis can result from traumatic injury to the skin that involves contamination with soil. After skin inoculation, a superficial abscess or localized cellulitis can develop. Co-trimoxazole is the drug of choice for all types of nocardiosis. We are reporting a case of Nocardia cyriacigeorgica presenting as cellulitis followed tooth extraction.

Characterization of multi-drug resistant Klebsiella pneumoniae isolates from urinary tract infected-women in Sylhet city, Bangladesh

Aims@#Klebsiella pneumoniae is considered to be one of the most frequent bacterial species associated with urinary tract infections (UTIs) and recurrent UTIs (RUTIs) worldwide. The present study aimed to comprehensively characterize K. pneumoniae isolates from women suffering from UTI and RUTIs. @*Methodology and results@#A total of 15 clinical isolates, collected from different hospitals in Bangladesh, were tested for biochemical features, and amplified by PCR. Antibiogram assay was performed by disk-diffusion assay. Phylogenetic and functional features were analyzed using bioinformatics platform. XLSTAT was used for principal component analysis (PCA). PCR amplification using Klebsiella hemolysin gene (khe) confirmed the presence of K. pneumoniae in agarose gel with expected product size of 486 kb. Antibiogram assay revealed all K. pneumoniae isolates to be completely resistant to six out of ten relevant drugs namely ampicillin, cephradine, chloramphenicol, erythromycin, kanamycin and sulfamethoxazole used for treating UTIs in Bangladesh. Sequencing of 16S rRNA gene of clinically significant K. pneumoniae isolates showed a high level of sequence divergence among the isolates from UTI and RUTIs as well as functional features such as SNP variants and restriction sites. @*Conclusion, significance and impact of study@#We surmise that the results could be used as a pipeline for further research in the identification of K. pneumoniae associated with UTI and RUTIs, and treatment of infection.

Globicatella sanguinis Bacteremia in a Korean Patient / 대한임상미생물학회지

Globicatella sanguinis is an unusual pathogen causing bacteremia, meningitis, and urinary tract infection, and can be misidentified as Streptococcus pneumoniae or viridans streptococci due to its colonial morphology. A 76-year-old female patient with hypertension and degenerative arthritis was admitted to the hospital complaining of knee joint pain. Blood culture revealed the presence of Gram-positive cocci, and the isolated organism was equally identified as S. pneumoniae using the MicroScan identification system (Beckman Coulter, USA) and Vitek 2 identification system (bioMérieux, USA). However, the isolate showed optochin resistance based on the optochin disk susceptibility test. The organism was finally confirmed to be G. sanguinis based on 16S rRNA sequencing and hydrogen sulfide production testing. Accurate identification of G. sanguinis isolated from aseptic body fluids including blood is important for appropriate antibiotic selection based on accurate application of interpretative criteria of antimicrobial susceptibility test.

A Case of Streptococcus gallolyticus subsp. gallolyticus Bacteremia in Liver Cirrhosis

Streptococcus bovis bacteremia in humans has been traditionally associated with infective endocarditis, colorectal cancer, and liver cirrhosis. S. bovis strains were previously categorized by biotype, but since the 2000s, they have been reclassified by DNA homology. We report a case of S. gallolyticus subsp. gallolyticus bacteremia, identified by 16S rRNA sequencing, in a patient diagnosed with liver cirrhosis. A 61-yr-old man with a history of liver cirrhosis presented to the hospital with a complaint of fever. Blood culture revealed the presence of gram-positive cocci, and the isolated organism was identified as S. bovis by the MicroScan identification kit (Beckman Coulter, USA), but as Enterococcus saccharolyticus by the Vitek 2 identification kit (bioMérieux, USA). The organism was finally confirmed as S. gallolyticus subsp. gallolyticus by 16S rRNA sequencing.

Five Cases of the Gram Variable Bacterium Paenibacillus urinalis Isolated from Clinical Specimens and its Clinical Significance

Paenibacillus urinalis was first isolated from the urine of a woman in 2008, and was reported to be a contaminant. Here, we report 5 cases of P. urinalis isolated over 5 months at a tertiary hospital. Using an API kit, 4 cases were classified as Cellulomonas species. Owing to the low reliability of API kit results and Gram stain results indicating gram variable bacilli for few specimens, MALDI-TOF MS and 16S rRNA gene sequencing were performed for identification. The last case showed Gram variable bacilli, and therefore, based on previous experience, 16S rRNA gene base sequence analysis was carried out without an additional API kit. All isolated strains were confirmed to be P. urinalis, and were judged to be contaminants. As for Gram variable bacteria, the use of current biochemical identification systems may lead to misidentification as other bacteria, which may cause unnecessary or improper use of antibiotics. Moreover, whereas most of the Paenibacillus species are reported to be contaminants, some of them are being reported as sources of infection. Therefore, more accurate identification will be necessary in the future. Accordingly, it is expected that accurate identification of this genus will help clinical physicians make decisions regarding appropriate treatment and use of antibiotics.

Improvement of Bacillus strains by mutation for overproduction of exopolygalacturonases.

Pectinases, produced by microorganisms, have wide range application in food industry, textile processing, paper making, coffee and tea fermentation, etc. It accounts for 10% of the global industrial enzymes produced. The most important and widely used commercial pectinase polygalacturonase is produced by alkalophilic strains of Bacillus sp. and Streptomyces sp. Here, we explored 29 bacterial strains isolated from rotten mango samples for polygalacturonase production and selected 16 strains through preliminary screening by well-plate method for enzyme activity. The maximum zone of inhibition of pectin was observed up to 28 mm in diameter but one strain ZM11 was exhibiting no activity. Quantitative dinitrisalicylic acid (DNS) assay for polygalacturonase enzyme was also performed for the selected bacterial isolates. All the strains bestowed significant enzyme activity with the highest activity of 2.4 U/µL exhibited by strain ZM3 (P ≤0.05). Characterization of the isolates was performed using different biochemical tests which also confirmed the isolates as members of the genus Bacillus. Mutation was induced to the selected strains by UV light and acridine orange for different periods of time. Qualitative and quantitative assays of the mutant bacterial isolates showed that the enzyme activity increased to 4.62 U/µL which clearly indicated that induced mutation enhanced the ability of Bacillus strains to produce more polygalacturonase enzyme up to 3-fold as compared to the wild strains (P ≤0.05). Molecular characterization by 16S rRNA sequences further confirmed that the bacterial isolates belong to Bacillus subtilis and B. amyloliquefaciens.