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Cryptococcus gattii (Cg) is a facultative intracellular pathogen that can replicate and disseminate in mammalian macrophages, causing life-threatening cryptococcosis in both immunocompetent and immunocompromised individuals. Cryptococcus-macrophage interactions are crucial for cryptococcosis prognosis. However, the relationship between Cg pathogenicity and phagocytosis by macrophages has not yet been investigated in depth. In this study, a series of in vitro and in vivo experiments were conducted to investigate the interaction between macrophages and Cg. Flow cytometry was used to detect the phagocytic phenotypes of the Cg strains within macrophages. Scanning electron microscopy, transmission electron microscopy, and immunofluorescence were used to observe phagocytosis and proliferation, respectively. Survival and lung fungal burden tests were also performed. Our results show that Cg cells display different phagocytosis phenotypes, which are independent of the molecular type. Within macrophages, the high phagocytosis phenotype (HP) strains obtain higher intracellular proliferation than the low phagocytosis phenotype (LP) strains. At the early stage of infection in vivo, HP-inducing permissive granulomas within the lungs seldom limit the dissemination of cryptococci. In addition, HP strains could inhibit the formation of M1-type macrophages, proliferate intracellularly and disseminate extracellularly, and cause hypoxia induced by mucus and acidic polysaccharide accumulation in pulmonary alveoli much earlier than LP strains in vivo. Our work reveals that Cg displays diverse interactions with macrophages, which may enhance our understanding of the pathogenicity of this life-threatening pathogen.

Abstract Background Several reports have associated Staphylococcus lugdunensis with the incidence of severe infection in humans; however, the frequency and prevalence of this microorganism and thus the propensity of its antimicrobial drug resistance is unknown in China. The objective of the current study was to determine the prevalence of Staphylococcus lugdunensis among six hundred and seventy non-replicate coagulase negative Staphylococcus (CoNS) isolates collected in a 12-month period from clinical specimens in the General Hospital of the People’s Liberation Army in Beijing, China. Results Five (0.7%) of the 670 isolates of CoNS were identified as S. lugdunensis. Whereas three isolates were resistant to erythromycin, clindamycin, and penicillin and carried the ermC gene and a fourth one was resistant to cefoxitin and penicillin and carried the mecA gene, one isolate was not resistant to any of the tested antimicrobials. Pulse field gel electrophoretic analysis did not reveal widespread epidemiological diversity of the different isolates. Conclusion Hence, even though S. lugdunensis may be yet unrecognized and undefined in China, it still might be the infrequent cause of infection and profound multi-drug resistance in the same population.

A 10:1 pooled test strategy on-site at an airport of China was pursued, resulting in increased test throughput, limited use of reagents, and increased testing efficiency without loss of sensitivity. This testing approach has the potential to reduce the need for contact tracing when the results are delivered first time. [ABSTRACT FROM AUTHOR]

Objective: Hypervirulent Klebsiella pneumoniae (hvKP) has been characterized as the major pathogen associated with the development of liver abscesses. Although the prevalence of K. pneumoniae in liver abscesses in Chinese patients is on the rise, methods for the early laboratory identification and recognition of hvKP are lacking, and hvKP detection and surveillance are thus limited. Methods: A retrospective study was conducted in 240 patients with cultures that were positive for K. pneumoniae , who were hospitalized in the Hospital of Beijing area from May 2013 to August 2014. The clinical and molecular characteristics of K. pneumoniae strains that cause liver abscesses were studied. Thirty-eight (16.7%) of 240 patients positive for K. pneumoniae cultures had liver abscesses. Results: Among these K. pneumoniae isolates, 92.1% (35/38) and 84.2% (32/38) were rmpA and aerobactin (abc) positive, respectively, as indicated by multiplex PCR, while 81.6% (31/38) were string test positive. Intriguingly, K. pneumoniae isolates that were positive for rmpA, abc , or string test accounted for almost all isolates (97.3%; n = 37/38) and thus were defined as hvKP in this study. Based on the new criteria for hvKP, none of these hvKP strains were positive for extended spectrum β-lactamases. Conclusions: Compared to the string test alone, integrating rmpA and abc detection with the string test is robuster and more extensive in the identification of liver abscess-causing hvKP strains. Multiplex PCR assays may therefore accelerate the early laboratory detection of hvKP. [ABSTRACT FROM AUTHOR]

Background: The emergence and rapid spreading of multidrug-resistant Acinetobacter baumannii strains has become a major health threat worldwide. To better understand the genetic recombination related with the acquisition of drug-resistant elements during bacterial infection, we performed complete genome analysis on three newly isolated multidrug-resistant A. baumannii strains from Beijing using next-generation sequencing technology. Methodologies/Principal Findings: Whole genome comparison revealed that all 3 strains share some common drug resistant elements including carbapenem-resistant blaOXA-23 and tetracycline (tet) resistance islands, but the genome structures are diversified among strains. Various genomic islands intersperse on the genome with transposons and insertions, reflecting the recombination flexibility during the acquisition of the resistant elements. The blood-isolated BJAB07104 and ascites-isolated BJAB0868 exhibit high similarity on their genome structure with most of the global clone II strains, suggesting these two strains belong to the dominant outbreak strains prevalent worldwide. A large resistance island (RI) of about 121-kb, carrying a cluster of resistance-related genes, was inserted into the ATPase gene on BJAB07104 and BJAB0868 genomes. A 78-kb insertion element carrying tra-locus and blaOXA-23 island, can be either inserted into one of the tniB gene in the 121-kb RI on the chromosome, or transformed to conjugative plasmid in the two BJAB strains. The third strains of this study, BJAB0715, which was isolated from spinal fluid, exhibit much more divergence compared with above two strains. It harbors multiple drug-resistance elements including a truncated AbaR-22-like RI on its genome. One of the unique features of this strain is that it carries both blaOXA-23 and blaOXA-58 genes on its genome. Besides, an Acinetobacter lwoffii adeABC efflux element was found inserted into the ATPase position in BJAB0715. Conclusions: Our comparative analysis on currently completed Acinetobacter baumannii genomes revealed extensive and dynamic genome organizations, which may facilitate the bacteria to acquire drug-resistance elements into their genomes. [ABSTRACT FROM AUTHOR]

Cryptococcus gattii (Cg) is a facultative intracellular pathogen that can replicate and disseminate in mammalian macrophages, causing life-threatening cryptococcosis in both immunocompetent and immunocompromised individuals. Cryptococcus- macrophage interactions are crucial for cryptococcosis prognosis. However, the relationship between Cg pathogenicity and phagocytosis by macrophages has not yet been investigated in depth. In this study, a series of in vitro and in vivo experiments were conducted to investigate the interaction between macrophages and Cg. Flow cytometry was used to detect the phagocytic phenotypes of the Cg strains within macrophages. Scanning electron microscopy, transmission electron microscopy, and immunofluorescence were used to observe phagocytosis and proliferation, respectively. Survival and lung fungal burden tests were also performed. Our results show that Cg cells display different phagocytosis phenotypes, which are independent of the molecular type. Within macrophages, the high phagocytosis phenotype (HP) strains obtain higher intracellular proliferation than the low phagocytosis phenotype (LP) strains. At the early stage of infection in vivo , HP-inducing permissive granulomas within the lungs seldom limit the dissemination of cryptococci. In addition, HP strains could inhibit the formation of M1-type macrophages, proliferate intracellularly and disseminate extracellularly, and cause hypoxia induced by mucus and acidic polysaccharide accumulation in pulmonary alveoli much earlier than LP strains in vivo . Our work reveals that Cg displays diverse interactions with macrophages, which may enhance our understanding of the pathogenicity of this life-threatening pathogen.

Objective: To summarize the clinical and bacterial features of Klebsiella pnuemoniae liver abscess (KPLA) in order to provide the basis for the diagnosis and treatment of KPLA., Methods: Retrospective study was conducted. One hundred and fifty-two medical records, from 3 teaching hospitals in Beijing, between January 2010 and December 2014, were collected. Among which 137 complete medical records were analyzed. String test was carried out to detect the hypermucoviscosity phenotype. PCR was performed to check the capsular serotype and the virulent genes. Disk diffusion method was operated to obtain the antimicrobial resistance rates. The results were analyzed by chi-square test., Results: KPLA occurred mostly in middle-aged, male and diabetes mellitus patients. 92.7% (127/137) patients had fever. 80.3% (110/137) of the KLPA were single abscess, among which 80.9% (89/110) were in the right lobe and 33.6% (46/137) had air cavities.74.5% (102/137) of the white blood cell count, 83.2% (114/137) of the neutrophils' percentage, 78.1% (107/137) of alanine aminotransferase and 51.8% (71/137) of the total billrubin were elevated. 87.5% (133/152) of the Klebsiella pnuemoniae (Kpn) appeared to be hypermucoviscous, K1 was the most popular serotype, the second was K2, and the positive rates of virulent genes rmpA and aerobactin were 82.9% (126/152) and 88.2% (134/152), respectively. Among the isolates from the KPLA without other hepatobiliary diseases, the portion of K1 serotype, the positive rates of rmpA and aerobactin were 65.7%, 94.9% and 96.0%, respectively, higher than those of the 28.9%, 50.0% and 68.4% from the KPLA with other hepatobiliary diseases, while the undefined serotype potion was lower (5.1% vs 26.3%), the differences were statistically significant (χ(2)=14.98, 38.40, 17.61, 10.65, all P<0.01). Most of Kpn were susceptible to antimicrobials., Conclusions: KPLA has certain clinical features, and are mostly caused by hypervirulent isolates that are hypermucoviscous with rmpA and aerobactin genes. Most of the isolates are susceptible to antimicrobials.