Kingella kingae Septic Arthritis in an Older-Than-Expected Child.

Kingella kingae typically causes musculoskeletal infection in young children between the ages of 6 months and 4 years who may be in close contact with other similarly aged children who are colonized with the organism in their oropharynx. Kingella infections have rarely been described in older individuals with chronic medical conditions or immune compromise. This is a case report of a healthy, older child who developed an invasive infection due to Kingella kingae. Clinical and laboratory details are provided of an otherwise healthy 11-year-old female who developed an acute onset of septic arthritis of her shoulder. The organism was identified by culture and 16S polymerase chain reaction. Her clinical course necessitated an antibiotic change after the organism was correctly identified. The affected child had close contact with a 2-year-old sibling who recently had a viral upper respiratory infection. This case illustrates the potential for Kingella kingae to rarely cause invasive infection in older, healthy children. Supplemental laboratory techniques may be helpful to identify this organism. Although it is reasonable to limit the antibiotic spectrum for older children, clinicians should be aware of this possibility, particularly if there is a history of close contact with young children.

Resurgence of Rotavirus Genotype G12 in St. Louis During the 2014-2015 Rotavirus Season.

BACKGROUND: Rotaviruses are a leading cause of gastroenteritis. Rotavirus vaccination has dramatically reduced rotavirus occurrence; however, we have noticed mild to moderate recurrences in the St. Louis area in alternate years. In 2013, we found rotavirus genotype G12 to be the dominant strain in the St. Louis region. In this study, we again determined the distribution of genotypes and ascertained vaccine history in patients infected with rotavirus G12 during the 2014-15 season. METHODS: Samples submitted to the St. Louis Children's Hospital Microbiology Laboratory were tested for rotavirus using an antigen assay. We determined the VP7 genotype using amplicon sequence analysis. We determined genome sequences using high-throughput sequencing. We evaluated rotavirus immunization records when available. RESULTS: Of 30 typed viruses from 2014-15, 29 were G12 (97%). Whole-genome sequencing revealed few changes from G12 viruses analyzed in 2012-13. VP4 and VP7 sequences were >99% identical to previously sequenced G12 strains from St. Louis, and immune epitopes were conserved. Vaccination histories were available from 17 patients. Of these, 4 had been vaccinated, 3 had received incomplete vaccination or had a vaccine history that could not be confirmed, and 10 had not been vaccinated. CONCLUSIONS: G12 re-emerged as the predominant rotavirus genotype in 2014-15, comprising a higher percentage of cases than in 2012-13. The majority of patients with G12 and available vaccination histories were unvaccinated. There was no genomic evidence to indicate that the G12 strains in St. Louis had evolved to escape vaccine protection. Our work emphasizes the need for continued surveillance.

Primary osteomyelitis caused by hypervirulent Klebsiella pneumoniae.

Klebsiella pneumoniae is the most clinically relevant species of this genus, known to cause both community-acquired and nosocomial infections worldwide. In the past two decades, a distinct hypervirulent strain of K pneumoniae, characterised by its hypermucoviscous phenotype, has emerged as a clinically significant pathogen responsible for highly invasive infections. We present a case of osteomyelitis due to hypervirulent K pneumoniae reported in the USA. Genomic testing of the K pneumoniae isolate was performed due to the striking clinical presentation of the infection as well as the hypermucoid nature of the isolates, raising the suspicion for possible infection with the hypervirulent strain. Whole-genome sequencing and additional PCR testing demonstrated the isolate to be a K1 serotype, sequence type 23 strain expressing rmpA and rmpA2. Given the multiple reports of this pathogen causing invasive infections, clinicians should be aware of the possible presentation of metastatic and severe infection, including osteomyelitis, due to the hypervirulent strain of K pneumoniae not typical of classic K pneumoniae variants. In this Grand Round, we review the clinical features of hypervirulent K pneumoniae and its link to invasive infections, and discuss the need for improved awareness and identification of the pathogen.

The DNA Sensor AIM2 Maintains Intestinal Homeostasis via Regulation of Epithelial Antimicrobial Host Defense.

Microbial pattern molecules in the intestine play immunoregulatory roles via diverse pattern recognition receptors. However, the role of the cytosolic DNA sensor AIM2 in the maintenance of intestinal homeostasis is unknown. Here, we show that Aim2(-/-) mice are highly susceptible to dextran sodium sulfate-induced colitis that is associated with microbial dysbiosis as represented by higher colonic burden of commensal Escherichia coli. Colonization of germ-free mice with Aim2(-/-) mouse microbiota leads to higher colitis susceptibility. In-depth investigation of AIM2-mediated host defense responses reveals that caspase-1 activation and IL-1ß and IL-18 production are compromised in Aim2(-/-) mouse colons, consistent with defective inflammasome function. Moreover, IL-18 infusion reduces E. coli burden as well as colitis susceptibility in Aim2(-/-) mice. Altered microbiota in inflammasome-defective mice correlate with reduced expression of several antimicrobial peptides in intestinal epithelial cells. Together, these findings implicate DNA sensing by AIM2 as a regulatory mechanism for maintaining intestinal homeostasis.

The NLRP3 inflammasome functions as a negative regulator of tumorigenesis during colitis-associated cancer.

Colitis-associated cancer (CAC) is a major complication of inflammatory bowel diseases. We show that components of the inflammasome are protective during acute and recurring colitis and CAC in the dextran sulfate sodium (DSS) and azoxymethane + DSS models. Mice lacking the inflammasome adaptor protein PYCARD (ASC) and caspase-1 demonstrate increased disease outcome, morbidity, histopathology, and polyp formation. The increased tumor burden is correlated with attenuated levels of IL-1beta and IL-18 at the tumor site. To decipher the nucleotide-binding domain, leucine-rich-repeat-containing (NLR) component that is involved in colitis and CAC, we assessed Nlrp3 and Nlrc4 deficient mice. Nlrp3(-/-) mice showed an increase in acute and recurring colitis and CAC, although the disease outcome was less severe in Nlrp3(-/-) mice than in Pycard(-/-) or Casp1(-/-) animals. No significant differences were observed in disease progression or outcome in Nlrc4(-/-) mice compared with similarly treated wild-type animals. Bone marrow reconstitution experiments show that Nlrp3 gene expression and function in hematopoietic cells, rather than intestinal epithelial cells or stromal cells, is responsible for protection against increased tumorigenesis. These data suggest that the inflammasome functions as an attenuator of colitis and CAC.

Identification of functional Tat signal sequences in Mycobacterium tuberculosis proteins.

The twin-arginine translocation (Tat) pathway is a system used by some bacteria to export proteins out from the cytosol to the cell surface or extracellular environment. A functional Tat pathway exists in the important human pathogen Mycobacterium tuberculosis. Identification of the substrates exported by the Tat pathway can help define the role that this pathway plays in the physiology and pathogenesis of M. tuberculosis. Here we used a reporter of Tat export, a truncated beta-lactamase, 'BlaC, to experimentally identify M. tuberculosis proteins with functional Tat signal sequences. Of the 13 proteins identified, one lacks the hallmark of a Tat-exported substrate, the twin-arginine dipeptide, and another is not predicted by in silico analysis of the annotated M. tuberculosis genome. Full-length versions of a subset of these proteins were tested to determine if the native proteins are Tat exported. For three proteins, expression in a Deltatat mutant of Mycobacterium smegmatis revealed a defect in precursor processing compared to expression in the wild type, indicating Tat export of the full-length proteins. Conversely, two proteins showed no obvious Tat export in M. smegmatis. One of this latter group of proteins was the M. tuberculosis virulence factor phospholipase C (PlcB). Importantly, when tested in M. tuberculosis a different result was obtained and PlcB was exported in a twin-arginine-dependent manner. This suggests the existence of an M. tuberculosis-specific factor(s) for Tat export of a proven virulence protein. It also emphasizes the importance of domains beyond the Tat signal sequence and bacterium-specific factors in determining if a given protein is Tat exported.

Genetic basis of murine antibacterial defense to streptococcal lung infection.

To evaluate the effect of genetic background on antibacterial defense to streptococcal infection, eight genetically diverse strains of mice (A/J, DBA/2J, CAST/Ei, FVB/NJ, BALB/cJ, C57BL/6J, 129/SvImJ, and C3H/HeJ) and tlr2-deficient mice (C57BL/6(tlr2-/-)) were infected with three doses of Streptococcus zooepidemicus (500, 5,000, or 50,000 colony-forming units) by alveolar challenge. There was a range of susceptibility between the strains at each dose and time point (6, 24, and 96 h). At the lowest dose, the 129/SvImJ and C3H/HeJ strains had significantly higher bacterial counts at all time points after infection, when compared to A/J, DBA/2J, CAST/Ei, FVB/NJ, which were resistant to infection at the low dose of innoculum. At the medium dose, 129/SvImJ and C3H/HeJ had higher bacterial counts, while A/J, DBA/2J, and BALB/cJ showed reduced streptococcal growth. After the highest dose of Streptococcus, there were minimal differences between strains, suggesting the protective impact of modifier genes can be overcome. TLR2-deficient animals contained increased bacterial load with reduced cytokines after 96 h when compared to C57BL/6J controls suggesting a role of innate immunity in late antibacterial defense. Overall, we identify vulnerable (129/SvlmJ and C3H/HeJ) and resistant (A/J, FVB, and DBA) mouse strains to streptococcal lung infection, which demonstrate divergent genetic expression profiles. These results demonstrate that innate differences in pulmonary host defense to S. zooepidemicus are dependent on host genetic factors.