Surveillance for norovirus and enteric bacterial pathogens as etiologies of acute gastroenteritis at U.S. military recruit training centers, 2011-2016.

An estimated 179 million cases of acute gastroenteritis (AGE) occur each year in the U.S. and AGE is commonly reported within both training and deployed U.S. military populations. Beginning in 2011, the Operational Infectious Diseases laboratory at Naval Health Research Center (NHRC) has undertaken routine surveillance of four U.S. military training facilities to systematically track the prevalence of AGE and to establish its etiologies among U.S. military recruits. Employing both molecular and standard microbiological techniques, NHRC routinely assays for pathogens of direct military relevance, including norovirus genogroups I and II, Salmonella, Shigella, and Campylobacter. During its initial surveillance efforts (2011-2016), NHRC identified norovirus as the primary etiology of both sporadic cases and outbreaks of AGE among trainees.

Peritraumatic Behavior Questionnaire - Observer Rated: Validation of the objective version of a measure for combat-related peritraumatic stress.

AIM: To validate the first third-person-rated measure assessing combat-related peritraumatic stress symptoms and evaluate its psychometric properties and war-zone applicability. METHODS: The valid assessment of peritraumatic symptoms in the theater of military operations represents a significant challenge in combat-related, mental health research, which mainly relies on retrospective, subjective self-report ratings. This longitudinal observational study used data from actively deployed troops to correlate third-person observer ratings of deployment peritraumatic behaviors [Peritraumatic Behavior Questionnaire - Observer Rated (PBQ-OR)] collected on a bi-monthly basis with post-deployment (1-wk follow-up) ratings of the previously validated PBQ self-rate version (PBQ-SR), and (3-mo follow-up) clinician assessed and self-report posttraumatic stress disorder (PTSD) symptoms (Clinician Administered PTSD Scale, PTSD Checklist). Cronbach's alpha (α) and correlation coefficients were calculated to assess internal reliability and concurrent validity respectively. RESULTS: Eight hundred and sixty male Marines were included in this study after signing informed consents at pre-deployment (mean age 23.2 ± 2.6 years). Although our findings were limited by an overall sparse return rate of PBQ-OR ratings, the main results indicate satisfactory psychometric properties with good internal consistency for the PBQ-OR (α = 0.88) and high convergent and concurrent validity with 1-wk post-deployment PBQ-SR ratings and 3-mo posttraumatic stress symptoms. Overall, later PBQ-OR report date was associated with higher correlation between PBQ-OR and post-deployment measures. Kappa analysis between PBQ-OR and PBQ-SR single items, showed best agreement in questions relating of mortal peril, desire for revenge, and experience of intense physical reactions. Logistic regression demonstrated satisfactory predictive validity of PBQ-OR total score with respect to PTSD caseness (OR = 1.0513; 95%CI: 1.011-1.093; P = 0.02). CONCLUSION: Since no comparable tools have been developed, PBQ-OR could be valuable as real-time screening tool for earlier detection of Service Members at risk.

Molecular Epidemiology of Adenovirus Type 21 Respiratory Strains Isolated From US Military Trainees (1996-2014).

BACKGROUND: The circulation of human adenovirus type 21 (HAdV21) in the United States has been documented since the 1960s in association with outbreaks of febrile respiratory illness (FRI) in military boot camps and civilian cases of respiratory disease. METHODS: To describe the molecular epidemiology of HAdV21 respiratory infections across the country, 150 clinical respiratory isolates obtained from continuous surveillance of military recruit FRI, and 23 respiratory isolates recovered from pediatric and adult civilian cases of acute respiratory infection were characterized to compile molecular typing data spanning 37 years (1978-2014). RESULTS: Restriction enzyme analysis and genomic sequencing identified 2 clusters of closely related genomic variants readily distinguishable from the prototype and designated 21a-like and 21b-like. A-like variants predominated until 1999. A shift to b-like variants was noticeable by 2007 after a 7-year period (2000-2006) of cocirculation of the 2 genome types. US strains are phylogenetically more closely related to European and Asian strains isolated over the last 4 decades than to the Saudi Arabian prototype strain AV-1645 isolated in 1956. CONCLUSIONS: Knowledge of circulating HAdV21 variants and their epidemic behavior will be of significant value to local and global FRI surveillance efforts.

Effect of multiple, simultaneous vaccines on polio seroresponse and associated health outcomes.

BACKGROUND: Administration of multiple simultaneous vaccines to infants, children, and military recruits is not uncommon. However, little research exists to examine associated serological and health effects, especially in adults. METHOD: We retrospectively examined 416 paired serum specimens from U.S. military subjects who had received the inactivated polio vaccine (IPV) alone or in combination with either 1 other vaccine (<3 group) or 4 other vaccines (>4 group). Each of the 2 groups was subdivided into 2 subgroups in which Tdap was present or absent. RESULTS: The >4 group was associated with a higher proportion of polio seroconversions than the <3 group (95% vs. 58%, respectively, p<0.01). Analysis of the <3 subgroup that excluded Tdap vs. the >4 subgroup that excluded Tdap showed no difference between them (p>0.1). However, the >4 subgroup that included Tdap had significantly more seroconversions than either the <3 subgroup that excluded Tdap or the >4 subgroup that excluded Tdap (p<0.01). Overall, at least 98% of subjects were at or above the putative level of seroprotection both pre- and post-vaccination, yet at least 81% of subjects seroconverted. In an analysis of 400 of the subjects in which clinic in- and outpatient encounters were counted over the course of 1 year following vaccinations, there was no significant difference between the 2 groups (p>0.1). CONCLUSION: A combination of >4 vaccines including IPV appeared to have an immunopotentiation effect on polio seroconversion, and Tdap in particular was a strong candidate for an important role. The dose of IPV we studied in our subjects, who already had a high level of seroprotection, acted as a booster. In addition, there appear to be no negative health consequences from receiving few versus more multiple simultaneous vaccinations.

The cumulative effect of different childhood trauma types on self-reported symptoms of adult male depression and PTSD, substance abuse and health-related quality of life in a large active-duty military cohort.

History of childhood trauma (CT) is highly prevalent and may lead to long-term consequences on physical and mental health. This study investigated the independent association of CT with symptoms of adult depression and posttraumatic stress disorder (PTSD), mental and physical health-related quality of life (HRQoL), as well as current tobacco consumption and alcohol abuse in a large homogenous cohort of 1254 never-deployed, young male Marines enrolled in the Marine Resiliency Study. Independent effects of CT history, number and type of CT on outcomes were analyzed using hierarchical multivariate logistic regression models. Our results suggested dose-dependent negative effect of an increasing number of trauma types of CT on depression, PTSD and HRQoL. Experience of single CT type demonstrated overall weak effects, while history of multiple CT types distinctively increased the likelihood of adult PTSD symptomology (OR: 3.1, 95% CI: 1.5-6.2), poor mental (OR: 2.3, 95% CI: 1.7-3.1) and physical HRQoL (OR: 1.4, 95% CI: 1.1-1.9). Risk for depression symptoms was similar for both single and multiple CT (OR: 2.2, 95% CI: 1.3-3.8 and OR: 2.1, 95% CI: 1.2-3.5 respectively). CT history had no effects on current tobacco use and alcohol abuse. Our study thus provides evidence for substantial additive effect of different CT types on adult mental and physical health with increasing levels of exposure.

Broad-spectrum biosensor capable of detecting and identifying diverse bacterial and Candida species in blood.

We describe an assay which uses broad-spectrum, conserved-site PCR paired with mass spectrometry analysis of amplicons (PCR/electrospray ionization-mass spectrometry [ESI-MS]) to detect and identify diverse bacterial and Candida species in uncultured specimens. The performance of the assay was characterized using whole-blood samples spiked with low titers of 64 bacterial species and 6 Candida species representing the breadth of coverage of the assay. The assay had an average limit of detection of 100 CFU of bacteria or Candida per milliliter of blood, and all species tested yielded limits of detection between 20 and 500 CFU per milliliter. Over 99% of all detections yielded correct identifications, whether they were obtained at concentrations well above the limit of detection or at the lowest detectable concentrations. This study demonstrates the ability of broad-spectrum PCR/ESI-MS assays to detect and identify diverse organisms in complex natural matrices that contain high levels of background DNA.

Trends in meningococcal disease in the United States military, 1971-2010.

Meningococci have historically caused extensive illness among members of the United States military. Three successive meningococcal vaccine types were used from 1971 through 2010; overall disease incidence dropped by >90% during this period. During 2006-2010, disease incidence of 0.38 (cases per 100,000 person-years) among members of the US military was not significantly different from the incidence of 0.26 among the age-matched US general population. Of the 26 cases in the US military, 5 were fatal, 15 were vaccine failures (e.g., illness in a person who had been vaccinated), and 9 were caused by Neisseria meningitidis serogroup Y. Incidences among 17- to 19-year-old basic trainees and among US Marines were significantly higher than among comparison military populations (p<0.05). No apparent change in epidemiology of meningococcal disease was observed after replacement of quadrivalent polysaccharide vaccine with conjugate vaccine in 2007. The data demonstrate that vaccination with meningococcal vaccine is effective.

Factors associated with loss of penicillin G concentrations in serum after intramuscular benzathine penicillin G injection: a meta-analysis.

BACKGROUND: An interval of 3-4 weeks between intramuscular injections of 1.2 million units of benzathine penicillin G as prophylaxis against group A streptococcal infection is recommended by health organizations for patients with pediatric rheumatic fever and heart disease. METHODS: We reviewed the literature for evidence of the persistence of serum penicillin G during the first 4 weeks after the recommended dose of benzathine penicillin G. RESULTS: The weighted-mean concentration was <0.02 µg/mL by 3 weeks after the initial dose. Weighted means were lower in studies done after 1990 than before (P<0.01), in studies dealing with secondary versus primary prophylaxis (P<0.01) and in studies in children versus those in adults (P<0.02). CONCLUSIONS: Recommendations for benzathine penicillin G prophylaxis may need reevaluation.

Decreased serologic response in vaccinated military recruits during 2011 correspond to genetic drift in concurrent circulating pandemic A/H1N1 viruses.

BACKGROUND: Population-based febrile respiratory illness surveillance conducted by the Department of Defense contributes to an estimate of vaccine effectiveness. Between January and March 2011, 64 cases of 2009 A/H1N1 (pH1N1), including one fatality, were confirmed in immunized recruits at Fort Jackson, South Carolina, suggesting insufficient efficacy for the pH1N1 component of the live attenuated influenza vaccine (LAIV). METHODOLOGY/PRINCIPAL FINDINGS: To test serologic protection, serum samples were collected at least 30 days post-vaccination from recruits at Fort Jackson (LAIV), Parris Island (LAIV and trivalent inactivated vaccine [TIV]) at Cape May, New Jersey (TIV) and responses measured against pre-vaccination sera. A subset of 78 LAIV and 64 TIV sera pairs from recruits who reported neither influenza vaccination in the prior year nor fever during training were tested by microneutralization (MN) and hemagglutination inhibition (HI) assays. MN results demonstrated that seroconversion in paired sera was greater in those who received TIV versus LAIV (74% and 37%). Additionally, the fold change associated with TIV vaccination was significantly different between circulating (2011) versus the vaccine strain (2009) of pH1N1 viruses (ANOVA p value = 0.0006). HI analyses revealed similar trends. Surface plasmon resonance (SPR) analysis revealed that the quantity, IgG/IgM ratios, and affinity of anti-HA antibodies were significantly greater in TIV vaccinees. Finally, sequence analysis of the HA1 gene in concurrent circulating 2011 pH1N1 isolates from Fort Jackson exhibited modest amino acid divergence from the vaccine strain. CONCLUSIONS/SIGNIFICANCE: Among military recruits in 2011, serum antibody response differed by vaccine type (LAIV vs. TIV) and pH1N1 virus year (2009 vs. 2011). We hypothesize that antigen drift in circulating pH1N1 viruses contributed to reduce vaccine effectiveness at Fort Jackson. Our findings have wider implications regarding vaccine protection from circulating pH1N1 viruses in 2011-2012.

Serum penicillin G levels are lower than expected in adults within two weeks of administration of 1.2 million units.

When introduced in the 1950s, benzathine penicillin G (BPG) was shown to be effective in eradicating group A beta-hemolytic streptococcus (GAS) for at least 3 weeks after administration. Several studies since the 1990s suggest that at 3-4 weeks serum penicillin G levels are less than adequate (below MIC(90) of 0.016 µg/ml). We studied these levels for 4 weeks after the recommended dose of BPG in military recruits, for whom it is used as prophylaxis against GAS. The 329 subjects (mean age 20 years) each received 1.2 million units BPG IM and gave sera 1 day post injection and twice more at staggered time points over 4 weeks. Serum penicillin G levels were measured by liquid chromatography/tandem mass spectometry. The half-life of serum penicillin G was 4.1 days. By day 11, mean levels were <0.02 µg/ml, and by day 15<0.01 µg/ml. Levels in more than 50% of the subjects were below 0.02 µg/ml on day 9, and <.01 µg/ml on day 16. There was no demonstrable effect of subject body-surface area nor of the four different lots of BPG used. These data indicate that in healthy young adults serum penicillin G levels become less than protective <2½ weeks after injection of 1.2 million units of BPG. The findings require serious consideration in future medical and public health recommendations for treatment and prophylaxis of GAS upper respiratory tract infections.

Local changes in rates of group A Streptococcus disease and antibiotic resistance are associated with geographically widespread strain turnover events.

This study addresses the effects of dynamic strain turnover and antibiotic prophylaxis on rates of group A Streptococcus (GAS) antibiotic resistance and disease. The authors analyzed the strain distributions, disease rates, and patterns of antibiotic resistance of 802 GAS isolates collected from 2002 through 2007. These samples were collected from patients with GAS infection symptoms at 10 military facilities. Macrolide resistance peaked at 25% during 2004, due to the geographically widespread dominance of a single resistant strain (M75). The resistant strain was not retained regardless of local patterns of macrolide use, and resistance rates decreased upon replacement of M75 with macrolide-susceptible strains. Disease rates were similarly correlated with dominance of specific M types. Statistical analysis revealed temporal correlations between strain distributions at multiple locations. Only the most common strains yielded enough data at multiple sites for statistically significant comparison of temporal fluctuations in dominance, but these (including M44, M3, M18, M118, and M6) all yielded highly significant temporal correlations of 90% or greater on yearly scales. As expected given the complexity and variability of strain distributions on shorter time scales, analysis on a monthly scale yielded lower degrees of positive correlation (31-62%), but in this case all significant correlations were still positive. Shifts in antibiotic resistance profiles and disease rates at specific sites appear to be associated with strain replacements happening on larger scales, independent of antibiotic use at individual sites.

Broad spectrum respiratory pathogen analysis of throat swabs from military recruits reveals interference between rhinoviruses and adenoviruses.

Military recruits experience a high incidence of febrile respiratory illness (FRI), leading to significant morbidity and lost training time. Adenoviruses, group A Streptococcus pyogenes, and influenza virus are implicated in over half of the FRI cases reported at recruit training center clinics, while the etiology of the remaining cases is unclear. In this study, we explore the carriage rates and disease associations of adenovirus, enterovirus, rhinovirus, Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis in military recruits using high-density resequencing microarrays. The results showed that rhinoviruses, adenoviruses, S. pneumoniae, H. influenzae, and N. meningitidis were widely distributed in recruits. Of these five agents, only adenovirus showed significant correlation with illness. Among the samples tested, only pathogens associated with FRI, such as adenovirus 4 and enterovirus 68, revealed strong temporal and spatial clustering of specific strains, indicating that they are transmitted primarily within sites. The results showed a strong negative association between adenoviral FRI and the presence of rhinoviruses in recruits, suggesting some form of viral interference.

Inference of antibiotic resistance and virulence among diverse group A Streptococcus strains using emm sequencing and multilocus genotyping methods.

BACKGROUND: Group A Streptococcus pyogenes (GAS) exhibits a high degree of clinically relevant phenotypic diversity. Strains vary widely in terms of antibiotic resistance (AbR), clinical severity, and transmission rate. Currently, strain identification is achieved by emm typing (direct sequencing of the genomic segment coding for the antigenic portion of the M protein) or by multilocus genotyping methods. Phenotype analysis, including critical AbR typing, is generally achieved by much slower and more laborious direct culture-based methods. METHODOLOGY/PRINCIPAL FINDINGS: We compare genotype identification (by emm typing and PCR/ESI-MS) with directly measured phenotypes (AbR and outbreak associations) for 802 clinical isolates of GAS collected from symptomatic patients over a period of 6 years at 10 military facilities in the United States. All independent strain characterization methods are highly correlated. This shows that recombination, horizontal transfer, and other forms of reassortment are rare in GAS insofar as housekeeping genes, primary virulence and antibiotic resistance determinants, and the emm gene are concerned. Therefore, genotyping methods offer an efficient way to predict emm type and the associated AbR and virulence phenotypes. CONCLUSIONS/SIGNIFICANCE: The data presented here, combined with much historical data, suggest that emm typing assays and faster molecular methods that infer emm type from genomic signatures could be used to efficiently infer critical phenotypic characteristics based on robust genotype: phenotype correlations. This, in turn, would enable faster and better-targeted responses during identified outbreaks of constitutively resistant or particularly virulent emm types.

Antibodies to squalene in US Navy Persian Gulf War veterans with chronic multisymptom illness.

Since the end of the 1991 Gulf War, there have been reports of unexplained, multisymptom illnesses afflicting veterans who consistently report more symptoms than do nondeployed veterans. One of the many possible exposures suspected of causing chronic multisymptom illnesses Gulf War veterans is squalene, thought to be present in anthrax vaccine. We examined the relationship between squalene antibodies and chronic symptoms reported by Navy construction workers (Seabees), n=579. 30.2% were deployers, 7.4% were defined as ill, and 43.5% were positive for squalene antibodies. We found no association between squalene antibody status and chronic multisymptom illness (p=0.465). The etiology of Gulf War syndrome remains unknown, but should not include squalene antibody status.

Exploration of the effectiveness of social distancing on respiratory pathogen transmission implicates environmental contributions.

BACKGROUND: In both military and civilian settings, transmission of respiratory pathogens may be due to person-to-person and environmental contributions. This possibility was explored in a military training setting, where rates of febrile respiratory illness (FRI) often reach epidemic levels. METHODS: Population size and FRI rates were monitored over 10 months in the units of 50-90 individuals. Some units were open to the influx of potentially infectious convalescents (hereafter referred to as "open units," and some were closed to such an influx (hereafter referred to as "closed units"). Virologic testing and polymerase chain reaction analysis were used to detect adenovirus on surface structures. RESULTS: The odds ratio (OR) associated with FRI in closed units, compared with open units, was 1.13 (95% confidence interval [CI], 0.99-1.28). The OR in units with a population greater than the median size, compared with units with a population lower than the median size was 1.38 (95% CI, 1.23-1.55). Between 5% and 9% of surface samples obtained from selected units harbored viable adenovirus. CONCLUSIONS: FRI rates were not reduced in units that were closed to potentially contagious individuals. These findings imply that the primary source of the pathogen is likely environmental rather than human, and they underscore what is known about other virus types. Diligence in identifying the relative roles of different transmission routes is suggested for civilian settings similar to those described in the current study.

A cluster of Legionella-associated pneumonia cases in a population of military recruits.

A Legionella cluster was identified through retrospective PCR analysis of 240 throat swab samples from X-ray-confirmed pneumonia cases. These were identified among young and otherwise healthy U.S. military recruits during population-based surveillance for pneumonia pathogens. Results were confirmed by sequence analysis. Cases clustered tightly, suggesting a local environmental etiology.

Asthma hospitalizations among US military personnel, 1994 to 2004.

BACKGROUND: Acute asthma attacks strike unpredictably and may lead to hospitalization in otherwise healthy individuals. The burden of asthma hospitalization on the US health care system has greatly interested health care workers, many of whom see the incidence of asthma as increasing. OBJECTIVES: To examine the annual incidence of hospitalization and the frequency of subsequent hospitalization for asthma among all active-duty US military personnel between 1994 and 2004 and to determine demographic and occupational risk factors of asthma hospitalization within this generally healthy US population. METHODS: Annual demographic and occupational data were combined with electronic hospitalization records for patients with a discharge diagnosis of asthma. Using Cox proportional hazard modeling, the authors investigated demographic and occupational risk factors for asthma hospitalization. RESULTS: Women, married persons, health care workers, enlisted personnel, US Army personnel, and older persons were found to have a significantly greater risk of asthma hospitalization. Yearly rates of hospitalization declined from 22.3 per 100,000 persons to 12.6 per 100,000 persons between 1994 and 2004. CONCLUSIONS: Although these data have some limitations, they suggest that the burden of asthma hospitalizations in the large, healthy population of US military personnel has declined during the last decade. The decrease in hospitalization potentially reflects improved outpatient management strategies.

Risky alcohol use, age at onset of drinking, and adverse childhood experiences in young men entering the US Marine Corps.

OBJECTIVE: To examine how childhood experiences relate to risky underage drinking. DESIGN: A survey study of men starting military training between June 11, 2002, and April 5, 2006. Multivariate logistic regression models compared risky drinkers with "all others" or with nonrisky drinkers; excluding nondrinkers. SETTING: Marine Corps Recruit Depot, San Diego, Calif. PARTICIPANTS: Forty-one thousand four hundred eighty-two men aged 18 to 20 years. Main Exposures Age at drinking onset; childhood emotional, physical, and sexual abuse; childhood emotional and physical neglect; and household alcohol abuse, mental illness, domestic violence, or divorce. MAIN OUTCOME MEASURES: Risky drinking identified by scoring responses to 3 questions about alcohol consumption. RESULTS: Of 41,482 young men, 6128 (14.8%) were identified as risky drinkers, 18,693 (45.1%) as nonrisky drinkers, and 16 661 (40.2%) as nondrinkers. Among drinkers, early initiation of alcohol use was strongly associated with risky drinking, with a 5.5-fold risk if age at onset of drinking was 13 years or younger. Other associated factors included tobacco use, rural or small hometown, higher education, motivation to join the military for travel or adventure or to leave problems at home, numerous close friends and relatives, household alcohol abuse or mental illness, and childhood sexual or emotional abuse. When the comparison group included nondrinkers, additional associated factors included childhood physical abuse and domestic violence. CONCLUSIONS: These analyses confirm previous findings on risks for alcohol misuse in young adults and quantify these risks in new, large, multivariable models, adding unique perspective from a population of young Marines. Public health efforts to decrease alcohol misuse may be effectively targeted by prevention of underage alcohol use, tobacco use, and childhood abuse.

Mefloquine use and hospitalizations among US service members, 2002-2004.

The safety of mefloquine has not been well described in military populations. This study used standard military databases for mefloquine prescriptions and hospitalizations to investigate mefloquine safety among US service members from 2002 through 2004. Mefloquine-prescribed and deployed personnel (N = 8,858) were compared with two reference groups. The reference groups comprised US service members who were not prescribed mefloquine and resided in Europe or Japan (N = 156,203) or had been otherwise deployed (N = 232,381). In comparison with active-duty US service members residing in Europe or Japan, mefloquine-prescribed service members were at statistically significant decreased hazard for any-cause hospitalization, as well as diseases of the respiratory and digestive systems, musculoskeletal system and connective tissue diseases, injuries and poisonings, ill-defined conditions, and mood disorders. These results suggest there is no association between mefloquine prescriptions and severe health effects, as measured by hospitalizations, across a wide range of outcomes.