Fractional Third and Fourth Dose of RTS,S/AS01 Malaria Candidate Vaccine: A Phase 2a Controlled Human Malaria Parasite Infection and Immunogenicity Study.

BACKGROUND: Three full doses of RTS,S/AS01 malaria vaccine provides partial protection against controlled human malaria parasite infection (CHMI) and natural exposure. Immunization regimens, including a delayed fractional third dose, were assessed for potential increased protection against malaria and immunologic responses. METHODS: In a phase 2a, controlled, open-label, study of healthy malaria-naive adults, 16 subjects vaccinated with a 0-, 1-, and 2-month full-dose regimen (012M) and 30 subjects who received a 0-, 1-, and 7-month regimen, including a fractional third dose (Fx017M), underwent CHMI 3 weeks after the last dose. Plasmablast heavy and light chain immunoglobulin messenger RNA sequencing and antibody avidity were evaluated. Protection against repeat CHMI was evaluated after 8 months. RESULTS: A total of 26 of 30 subjects in the Fx017M group (vaccine efficacy [VE], 86.7% [95% confidence interval [CI], 66.8%-94.6%]; P < .0001) and 10 of 16 in the 012M group (VE, 62.5% [95% CI, 29.4%-80.1%]; P = .0009) were protected against infection, and protection differed between schedules (P = .040, by the log rank test). The fractional dose boosting increased antibody somatic hypermutation and avidity and sustained high protection upon rechallenge. DISCUSSIONS: A delayed third fractional vaccine dose improved immunogenicity and protection against infection. Optimization of the RTS,S/AS01 immunization regimen may lead to improved approaches against malaria. CLINICAL TRIALS REGISTRATION: NCT01857869.

Invasive Apophysomyces variabilis infection in a burn patient.

Apophysomyces variabilis is an emerging fungal pathogen that can cause significant infections in immunocompetent patients. We report a case of A. variabilis invasive wound infection in a 21-year-old male after a self-inflicted burn injury.

PCR for rapid diagnosis of acute Q fever at a combat support hospital in Iraq.

Acute Q fever is occasionally seen in U.S. military service members deployed to Iraq. Diagnosis relies on serology, which is not available in the combat zone. Improved diagnostic modalities are needed. We performed a pilot study investigating whether Joint Biological Agent Identification and Diagnostic System (JBAIDS), a ruggedized, deployable polymerase chain reaction (PCR) platform, might be useful in the diagnosis of acute Q fever. Patients presenting to a Combat Support Hospital in Iraq with undifferentiated fever had blood drawn for Q fever PCR and these results were compared with serology. PCR was positive in 6 of 9 patients with acute Q fever by serology and negative in all 9 patients with negative serology. These results suggest that PCR using the JBAIDS platform could be of use in the diagnosis of Q fever in deployed settings. Further research into this modality is warranted.

Leadership Lessons for a New Battalion Surgeon.

Military medical corps officers often do not have experience with line units until after residency. This case demonstrates the importance of understanding the flow of information within an operational setting. The case also highlights the challenges of advocating for patients as a young officer and physician.

Methicillin-resistant Staphylococcus aureus in wound cultures recovered from a combat support hospital in Iraq.

BACKGROUND: Staphylococcus aureus infections complicate care of combat-related injuries and can independently result in skin and soft-tissue infections during deployments or training. Community-associated methicillin-resistant S. aureus (CA-MRSA) strains seem to produce severe disease but retain susceptibility to many oral antimicrobials. This study characterizes 84 MRSA isolates recovered from wound cultures at a combat support hospital in Iraq. METHODS: MRSA strains recovered from December 2007 through March 2009 were analyzed. Antimicrobial resistance testing was determined by broth microdilution and the BD Phoenix Automated Microbiology System. The genotypic pattern was analyzed by pulsed-field gel electrophoresis and polymerase chain reaction identification of resistance and virulence genes. RESULTS: No MRSA isolates from wound cultures were resistant to vancomycin. The most active oral antistaphylococcal agents were tetracycline (95% susceptibility), trimethoprim-sulfamethoxazole (94%), and clindamycin (94%). Of agents not typically recommended as monotherapy, 98% of isolates were susceptible to rifampin, 91% to moxifloxacin, and 60% to levofloxacin. The most common pulsed-field type (PFT) was USA300 (79%). The typical staphylococcal cassette chromosome mec IV elements carrying the CA-MRSA resistance genes were present in 88% of the isolates. Panton-Valentine leukocidin virulence genes were identified in 88% of isolates, including 100% of PFT USA300. The virulence gene associated with an arginine catabolic mobile element was present in 75% of isolates, including 94% of PFT USA300. CONCLUSION: This study is the first genotypic and phenotypic characterization of CA-MRSA recovered from wound cultures in a deployed combat hospital. The pattern noted was similar to that seen in soldiers stationed in the United States.

Presence and molecular epidemiology of virulence factors in methicillin-resistant Staphylococcus aureus strains colonizing and infecting soldiers.

Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as an important cause of skin and soft-tissue infections (SSTI). The understanding of the molecular epidemiology and virulence of MRSA continues to expand. From January 2005 to December 2005, we screened soldiers for MRSA nasal colonization, administered a demographic questionnaire, and monitored them prospectively for SSTI. All MRSA isolates underwent molecular analysis, which included pulsed-filed gel electrophoresis (PFGE) and PCR for Panton-Valentine leukocidin (PVL), the arginine catabolic mobile element (ACME), and the staphylococcal cassette chromosome mec (SCCmec). Of the 3,447 soldiers screened, 134 (3.9%) had MRSA colonization. Of the 3,066 (89%) who completed the study, 39 developed culture-confirmed MRSA abscesses. Clone USA300 represented 53% of colonizing isolates but was responsible for 97% of the abscesses (P < 0.001). Unlike colonizing isolates, isolates positive for USA300, PVL, ACME, and type IV SCCmec were significantly associated with MRSA abscess isolates. As determined by multivariate analysis, risk factors for MRSA colonization were a history of SSTI and a history of hospitalization. Although various MRSA strains may colonize soldiers, USA300 is the most virulent when evaluated prospectively, and PVL, ACME, and type IV SCCmec are associated with these abscesses.

Recovery of multidrug-resistant bacteria from combat personnel evacuated from Iraq and Afghanistan at a single military treatment facility.

U.S. combat casualties from Iraq and Afghanistan continue to develop infections with multidrug-resistant (MDR) bacteria. This study assesses the infection control database and clinical microbiology antibiograms at a single site from 2005 to 2007, a period when all Operation Iraqi Freedom (OIF)/Operation Enduring Freedom (OEF) casualties admitted to the facility underwent initial isolation and screening for MDR pathogens. During this 3-year period, there were 2,242 OIF/OEF admissions: 560 in 2005, 724 in 2006, and 958 in 2007. The most commonly recovered pathogens from OIF/OEF admission screening cultures were methicillin-resistant Staphylococcus aureus (MRSA), Klebsiella pneumoniae and Acinetobacter. The yearly nosocomial infection rate of these three pathogens among OIF/OEF admissions ranged between 2 and 4%. There were remarkable changes in resistance profiles for Acinetobacter, K. pneumoniae, and S. aureus over time. Despite aggressive infection control procedures, there is continued nosocomial transmission within the facility and increasing antimicrobial resistance in some pathogens. Novel techniques are needed to control the impact of MDR bacteria in medical facilities.

Association of bacterial colonization at the time of presentation to a combat support hospital in a combat zone with subsequent 30-day colonization or infection.

U.S. casualties have developed multidrug-resistant (MDR) bacterial infections. A surveillance project to evaluate U.S. military patients for the presence of MDR pathogens from wounding through the first 30 days of care in the military healthcare system (MHS) was performed. U.S. military patients admitted to a single combat support hospital in Iraq during June-July of 2007 had screening swabs obtained for the detection of MDR bacteria and a subsequent retrospective electronic medical records review for presence of colonization or infection in the subsequent 30 days. Screening of 74 U.S. military patients in Iraq found one colonized with methicillin-resistant Staphylococcus aureus. Fifty-six patients of these were screened for Acinetobacter in Germany and one found colonized. Of patients evacuated to the U.S., 9 developed infections. Carefully obtained screening cultures immediately after injury combined with look-back monitoring supports the role of nosocomial transmission. Consistent infection control strategies are needed for the entire MHS.

Antimicrobial susceptibilities of geographically diverse clinical human isolates of Leptospira.

Although antimicrobial therapy of leptospirosis has been studied in a few randomized controlled clinical studies, those studies were limited to specific regions of the world and few have characterized infecting strains. A broth microdilution technique for the assessment of antibiotic susceptibility has been developed at Brooke Army Medical Center. In the present study, we assessed the susceptibilities of 13 Leptospira isolates (including recent clinical isolates) from Egypt, Thailand, Nicaragua, and Hawaii to 13 antimicrobial agents. Ampicillin, cefepime, azithromycin, and clarithromycin were found to have MICs below the lower limit of detection (0.016 microg/ml). Cefotaxime, ceftriaxone, imipenem-cilastatin, penicillin G, moxifloxacin, ciprofloxacin, and levofloxacin had MIC(90)s between 0.030 and 0.125 microg/ml. Doxycycline and tetracycline had the highest MIC(90)s: 2 and 4 microg/ml, respectively. Doxycycline and tetracycline were noted to have slightly higher MICs against isolates from Egypt than against strains from Thailand or Hawaii; otherwise, the susceptibility patterns were similar. There appears to be possible variability in susceptibility to some antimicrobial agents among strains, suggesting that more extensive testing to look for geographic variability should be pursued.

Acinetobacter skin carriage among US army soldiers deployed in Iraq.

Skin carriage of Acinetobacter calcoaceticus-baumannii complex was not detected among a representative sample of 102 US Army soldiers stationed in Iraq. This observation refutes the hypothesis that preinjury skin carriage serves as the reservoir for the Acinetobacter infections seen in US military combat casualties.

Acinetobacter skin colonization of US Army Soldiers.

OBJECTIVE: To evaluate whether skin colonization with Acinetobacter calcoaceticus-baumannii complex exists in a population of healthy, nondeployed US Army soldiers and, if present, how it might relate to the infections seen in current war casualties. DESIGN: We sampled various skin sites of soldiers to test for the presence of A. calcoaceticus-baumannii complex and to establish the prevalence of colonization. We then used ribotyping and antimicrobial susceptibility profiles to compare the isolates we recovered with A. calcoaceticus-baumannii complex isolates from injured soldiers. SETTING: Fort Sam Houston, Texas. PARTICIPANTS: A population of healthy, nondeployed US Army soldiers in training. RESULTS: A total of 17% of healthy soldiers were found to harbor A. calcoaceticus-baumannii complex. However, the strains differed from those recovered from injured soldiers. CONCLUSIONS: Skin carriage of A. calcoaceticus-baumannii complex exists among soldiers before deployment. However, the difference in the strains isolated from healthy soldiers, compared with the strains from injured soldiers, makes it difficult to identify skin colonization as the source of infection.

Viability of Leptospira in BacT/ALERT MB media.

Recovery of Leptospira in the clinical setting is typically low as specialized culture media is needed. Previous data demonstrated that blood culture media commonly available to most clinical laboratories do not adequately sustain viable Leptospira. We hypothesized that mycobacterial blood culture medium, which is often readily available to most clinical laboratories, might be able to support the growth of Leptospira. Leptospires and fresh human blood were inoculated into BacT/ALERT (bioMérieux, Durham NC) mycobacterial (MB) and enriched mycobacterial bottles. Standard aerobic (FA) and anaerobic (SN) bottles were also inoculated as a control group. Inoculated bottles were then evaluated for their ability to support Leptospira growth using dark-field microscopy, subculture, and an automated growth detection system. Viable leptospires were detected in MB bottles up to day 14. FA and SN were performed in accordance with prior data. We conclude that MB and enriched MB bottles of the BacT/ALERT blood culture system can support viable leptospires.

Impact of Acinetobacter infection on the mortality of burn patients.

BACKGROUND: Acinetobacter calcoaceticus-baumannii complex (Acb) is recognized as an important cause of nosocomial infections. Although Acb can be associated with multidrug resistance, its impact on mortality in burn patients has not been fully elucidated. STUDY DESIGN: In a retrospective cohort study assessing medical records and microbiology laboratory data at a US military tertiary care burn center, we evaluated all patients admitted to the burn center between January 2003 and November 2005. Data collected included age, severity of burn, comorbidities, length of stay, and survival to hospital discharge. In addition, microbiology data were reviewed to determine which patients were infected with Acb during this time frame. These data were then used to compare patients infected with Acb to patients not infected. Multivariate analysis using logistic regression was performed to determine which patient characteristics were associated with increased mortality. RESULTS: There were 802 patients included in the study. Fifty-nine patients met the case definition for infection. An additional 52 patients were found to be colonized with Acb. Patients with Acb infection had more severe burns and comorbidities, and had longer lengths of stay compared with patients without Acb or those with Acb colonization. Mortality in infected patients was higher compared with those without infection (relative risk = 2.86, p = 0.001). On multivariate analysis, infection with Acb was not statistically associated with mortality. CONCLUSIONS: Multidrug-resistant Acb is a common cause of nosocomial infection in the burn patient population. Despite this, it does not independently affect mortality.

Antimicrobial activity against CA-MRSA and treatment of uncomplicated nonpurulent cellulitis.

UNLABELLED: Evaluation of: Pallin DJ, Binder WD, Allen MB et al. CLINICAL TRIAL: comparative effectiveness of cephalexin plus trimethoprim-sulfamethoxazole versus cephalexin alone for treatment of uncomplicated cellulitis: a randomized controlled trial. Clin. Infect. Dis. 56(12), 1754-1762 (2013). The rise of community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) has complicated the empirical antimicrobial treatment of cellulitis. CA-MRSA is frequently the cause of purulent infections, to include purulent cellulitis. The role of CA-MRSA in nonpurulent cellulitis is less clear. Published clinical practice guidelines suggest that CA-MRSA plays only a minor role in nonpurulent cellulitis and that initial treatment should be primarily directed at ß-hemolytic streptococci. Until now, there have been no data from prospective randomized control trials to support this recommendation. In this review, we examine the findings from a recent prospective, double-blind, randomized controlled trial that refutes the need for empirical coverage of CA-MRSA when treating nonpurulent cellulitis.

Evaluation of potential environmental contamination sources for the presence of multidrug-resistant bacteria linked to wound infections in combat casualties.

OBJECTIVE: To determine whether multidrug-resistant (MDR) gram-negative organisms are present in Afghanistan or Iraq soil samples, contaminate standard deployed hospital or modular operating rooms (ORs), or aerosolize during surgical procedures. DESIGN: Active surveillance. SETTING: US military hospitals in the United States, Afghanistan, and Iraq. METHODS: Soil samples were collected from sites throughout Afghanistan and Iraq and analyzed for presence of MDR bacteria. Environmental sampling of selected newly established modular and deployed OR high-touch surfaces and equipment was performed to determine the presence of bacterial contamination. Gram-negative bacteria aerosolization during OR surgical procedures was determined by microbiological analysis of settle plate growth. RESULTS: Subsurface soil sample isolates recovered in Afghanistan and Iraq included various pansusceptible members of Enterobacteriaceae, Vibrio species, Pseudomonas species, Acinetobacter lwoffii, and coagulase-negative Staphylococcus (CNS). OR contamination studies in Afghanistan revealed 1 surface with a Micrococcus luteus. Newly established US-based modular ORs and the colocated fixed-facility ORs revealed no gram-negative bacterial contamination prior to the opening of the modular OR and 5 weeks later. Bacterial aerosolization during surgery in a deployed fixed hospital revealed a mean gram-negative bacteria colony count of 12.8 colony-forming units (CFU)/dm(2)/h (standard deviation [SD], 17.0) during surgeries and 6.5 CFU/dm(2)/h (SD, 7.5; [Formula: see text]) when the OR was not in use. CONCLUSION: This study demonstrates no significant gram-negative bacilli colonization of modular and fixed-facility ORs or dirt and no significant aerosolization of these bacilli during surgical procedures. These results lend additional support to the role of nosocomial transmission of MDR pathogens or the colonization of the patient themselves prior to injury.

Efficacy of minocycline and tigecycline in a hamster model of leptospirosis.

Leptospirosis is a widespread zoonotic infection characterized by acute febrile illness. Severely ill patients may require empiric treatment with broad-spectrum antibiotics prior to definitive diagnosis. We evaluated the efficacy of minocycline and tigecycline against leptospirosis in a hamster model. Hamsters were treated with either minocycline (5, 10, or 25 mg/kg per day) or tigecycline (5, 10, or 25 mg/kg per day) for 5 days. Controls included untreated animals and doxycycline-treated animals (5 mg/kg per day). Nine days after infection, all untreated animals were dead. All treated hamsters survived to the end of study (day 21). Study groups showed significantly improved survival compared to the untreated group (P < .01). Minocycline and tigecycline showed survival benefit comparable to the standard treatment, doxycycline. In the absence of doxycycline, minocycline may be considered as an alternative, while tigecycline may be useful in the management of severely ill patients prior to a definitive diagnosis.

In vitro and in vivo activity of first generation cephalosporins against Leptospira.

Third generation cephalosporins are commonly used in the treatment of leptospirosis. The efficacy of first generation cephalosporins has been less well-studied. Susceptibility testing of 13 Leptospira strains (11 serovars) to cefazolin and cephalexin was conducted using broth microdilution. Median minimal inhibitory concentration (MIC) for cefazolin and cephalexin ranged from < 0.016 to 2 µg/mL (MIC(90) = 0.5 µg/mL) and from 1 to 8 µg/mL (MIC(90) = 8 µg/mL), respectively. Efficacy of cefazolin and cephalexin in an acute lethal hamster model of leptospirosis was studied. Survival rates for cefazolin were 80%, 100%, and 100%, and survival rates for cephalexin were 50%, 80%, and 100% (treated with 5, 25, and 50 mg/kg per day for 5 days, respectively). Each treatment group showed improved survival compared with no treatment (P < 0.01), and none of the therapies, regardless of dose, was statistically significantly different than doxycycline. These results support a potential role for first generation cephalosporins as alternative therapies for leptospirosis.