Paul C. F. Graf

Plasmodium vivax hospitalizations in a monoendemic malaria region: severe vivax malaria?

Severe malaria caused by Plasmodium vivax is no longer considered rare. To describe its clinical features, we performed a retrospective case control study in the subregion of Luciano Castillo Colonna, Piura, Peru, an area with nearly exclusive vivax malaria transmission. Severe cases and the subset of critically ill cases were compared with a random set of uncomplicated malaria cases (1:4). Between 2008 and 2009, 6,502 malaria cases were reported, including 106 hospitalized cases, 81 of which fit the World Health Organization definition for severe malaria. Of these 81 individuals, 28 individuals were critically ill (0.4%, 95% confidence interval = 0.2-0.6%) with severe anemia (57%), shock (25%), lung injury (21%), acute renal failure (14%), or cerebral malaria (11%). Two potentially malaria-related deaths occurred. Compared with uncomplicated cases, individuals critically ill were older (38 versus 26 years old, P < 0.001), but similar in other regards. Severe vivax malaria monoinfection with critical illness is more common than previously thought.

Natural Leishmania Infection of Lutzomyia auraensis in Madre de Dios, Peru, Detected by a Fluorescence Resonance Energy Transfer-Based Real-Time Polymerase Chain Reaction

Leishmania species of the Viannia subgenus are responsible for most cases of New World tegumentary leishmaniasis. However, little is known about the vectors involved in disease transmission in the Amazon regions of Peru. We used a novel real-time polymerase chain reaction (PCR) to assess Leishmania infections in phlebotomines collected in rural areas of Madre de Dios, Peru. A total of 1,299 non-blood fed female sand flies from 33 species were captured by using miniature CDC light traps. Lutzomyia auraensis was the most abundant species (63%) in this area. Seven of 164 pools were positive by PCR for Leishmania by kinetoplast DNA. The real-time PCR identified four Lu. auraensis pools as positive for L. (Viannia) lainsoni and L. (V.) braziliensis. The minimum infection prevalence for Lu. auraensis was estimated to be 0.6% (95% confidence interval = 0.20-1.42%). Further studies are needed to assess the importance of Lu. auraensis in the transmission of New World tegumentary leishmaniasis in hyperendemic areas of Peru.

A FRET-Based Real-Time PCR Assay to Identify the Main Causal Agents of New World Tegumentary Leishmaniasis

In South America, various species of Leishmania are endemic and cause New World tegumentary leishmaniasis (NWTL). The correct identification of these species is critical for adequate clinical management and surveillance activities. We developed a real-time polymerase chain reaction (PCR) assay and evaluated its diagnostic performance using 64 archived parasite isolates and 192 prospectively identified samples collected from individuals with suspected leishmaniasis enrolled at two reference clinics in Lima, Peru. The real-time PCR assay was able to detect a single parasite and provided unambiguous melting peaks for five Leishmania species of the Viannia subgenus that are highly prevalent in South America: L. (V.) braziliensis, L. (V.) panamensis, L. (V.) guyanensis, L. (V.) peruviana and L. (V.) lainsoni. Using kinetoplastid DNA-based PCR as a gold standard, the real-time PCR had sensitivity and specificity values of 92% and 77%, respectively, which were significantly higher than those of conventional tests such as microscopy, culture and the leishmanin skin test (LST). In addition, the real-time PCR identified 147 different clinical samples at the species level, providing an overall agreement of 100% when compared to multilocus sequence typing (MLST) data performed on a subset of these samples. Furthermore, the real-time PCR was three times faster and five times less expensive when compared to PCR - MLST for species identification from clinical specimens. In summary, this new assay represents a cost-effective and reliable alternative for the identification of the main species causing NWTL in South America.

Efficacy of Three Different Regimens of Primaquine for the Prevention of Relapses of Plasmodium vivax Malaria in the Amazon Basin of Peru

We evaluated the efficacy of three primaquine (PQ) regimes to prevent relapses with Plasmodium vivax through an open-label randomized trial in Loreto, Peru. Vivax monoinfections were treated with chloroquine for 3 days and PQ in three different regimes: 0.5 mg/kg per day for 5 days (150 mg total), 0.5 mg/kg per day for 7 days (210 mg total), or 0.25 mg/kg per day for 14 days (210 mg total). Biweekly fever assessments and bimonthly thick smears were taken for 210 days. Recurrences after 35 days were considered relapses. One hundred eighty cases were enrolled in each group; 90% of cases completed follow-up. There were no group-related differences in age, sex, or parasitemia. Relapse rates were similar in the 7- and 14-day regimes (16/156 = 10.3% and 22/162 = 13.6%, P = 0.361) and higher in the 5-day group (48/169 = 28.4%, P < 0.001 and P = 0.001, respectively). The 7-day PQ regimen used in Peru is as efficacious as the recommended 14-day regimen and superior to 5 treatment days.

Detection of Rickettsia parkeri from within Piura, Peru, and the First Reported Presence of Candidatus Rickettsia andeanae in the Tick Rhipicephalus sanguineus

Domestic farm animals (n=145) were sampled for the presence of ectoparasites in northwestern Peru during March, 2008. Ninety domestic animals (62%) were positive for the presence of an ectoparasite(s) and produced a total collection of the following: 728 ticks [Amblyomma maculatum, Anocentor nitens, Rhipicephalus (Boophilus) microplus, Rhipicephalus sanguineus, and Otobius megnini], 12 lice (Haematopinus suis), and 3 fleas (Ctenocephalides felis). A Rickettsia genus-specific qPCR assay was performed on nucleic acid preparations of the collected ectoparasites that resulted in 5% (37/743, 35 ticks and 2 fleas) of the ectoparasites positive for the presence of Rickettsia. DNA from the positive individual ticks was tested with 2 other qPCR assays for the presence of the ompB gene in Candidatus Rickettsia andeanae or Rickettsia parkeri. Candidatus R. andeanae was found in 25 A. maculatum ticks and in two Rh. sanguineus ticks, whereas R. parkeri was detected in 6 A. maculatum ticks. Two A. maculatum were co-infected with both Candidatus R. andeanae and R. parkeri. Rickettsia felis was detected in 2 fleas, Ctenocephalides felis, by multilocus sequence typing of the 17-kD antigen and ompA genes. These findings expand the geographic range of R. parkeri to include Peru as well as expand the natural arthropod vector of Candidatus R. andeanae to include Rhipicephalus sanguineus.

Needs, Acceptability, and Value of Humanitarian Medical Assistance in Remote Peruvian Amazon Riverine Communities

Much debate exists regarding the need, acceptability, and value of humanitarian medical assistance. We conducted a cross-sectional study on 457 children under 5 years from four remote riverine communities in the Peruvian Amazon and collected anthropometric measures, blood samples (1-4 years), and stool samples. Focus groups and key informant interviews assessed perspectives regarding medical aid delivered by foreigners. The prevalence of stunting, anemia, and intestinal parasites was 20%, 37%, and 62%, respectively. Infection with multiple parasites, usually geohelminths, was detected in 41% of children. The prevalence of intestinal parasites both individual and polyparasitism increased with age. Participants from smaller communities less exposed to foreigners expressed lack of trust and fear of them. However, participants from all communities were positive about foreigners visiting to provide health support. Prevalent health needs such as parasitic infections and anemia may be addressed by short-term medical interventions. There is a perceived openness to and acceptability of medical assistance delivered by foreign personnel.

Pott's Disease? AIDS-Associated Mycobacterium heckeshornense Spinal Osteomyelitis and Diskitis

ABSTRACT Acid-fast bacillus (AFB) spinal osteomyelitis in a patient with AIDS is often presumed to be caused by reactivated Mycobacterium tuberculosis. However, other AFB pathogens can mimic M. tuberculosis and, to ensure appropriate and adequate therapy, should be considered by clinicians. We present a case of aggressive spinal osteomyelitis caused by Mycobacterium heckeshornense in an AIDS patient; a review of the literature is also included.

Seroconversions to Rickettsiae in US Military Personnel in South Korea.

To the Editor: Infections with typhus group rickettsiae (TGR), spotted fever group rickettsiae (SFGR), and scrub typhus group orientiae (STGO) have been reported among persons in South Korea in increasing numbers over the past decade (1,2). During 2001–2011 in South Korea, 51,825 orientiae group infections were reported (mean incidence 9.95 cases/100,000 residents/year) (2). TGR (Rickettsia typhi), SFGR (R. akari, R. japonica, R. monacensis, and R. felis), and STGO (Orientia tsutsugamusi) have been identified in their arthropod vectors and reservoirs in northern provinces and at US military training facilities in South Korea (3–5). Currently, little data exist on the risk for rickettsioses and scrub typhus for US military deployed to South Korea. Thus, a retrospective serologic investigation to determine the level of exposure to rickettsiae among 9,303 military personnel deployed to South Korea was conducted. The study used de-identified predeployment and postdeployment serum samples made available from the Department of Defense Serum Repository (6). The study group consisted of men in combat-related jobs at US military training sites and military installations in South Korea during 1990–1995 while on active duty continuously for >1 year. This study protocol was reviewed and approved by the Naval Medical Research Command Institutional Review Board in compliance with all applicable federal regulations governing the protection of human subjects. Age range of the 9,303 soldiers in the study group was 17–52 (median 24) years. Most (99.6%) were stationed in Dongducheon, Yongtaeri, and Seoul, located in the Gyeonggi and Gangwon provinces in northern South Korea. Primary military occupation specialties were infantryman (58.8%), fighting vehicle infantryman (22.4%), indirect fire infantryman (12.3%), and heavy anti-armor weapons infantryman (6.5%). The soldiers’ postdeployment serum samples (n = 9,303) were screened at a dilution of 1:100 for IgG against TGR, SFGR, and STGO by using group-specific ELISA whole-cell antigen preparations from R. typhi Wilmington, R. conorii Morrocan, and a mixture of O. tsutsugamushi Karp, Kato, and Gilliam, respectively (7,8). TGR, SFGR, and STGO IgG ELISA titers (range 100–>6,400) were determined for screen-positive (net absorbance >0.500) postdeployment serum samples, and results were compared with matched predeployment serum samples. Samples with a net total absorbance >1.000 for serum dilutions 1:100, 1:400, 1:1,600, and 1:6,400 were considered titer positive. The inverse of the highest dilution of titer positive serum that produced a net absorbance >0.200 was determined to be the titer. Serum samples from laboratory animals infected with R. felis reacted specifically in the SFGR ELISA but not in the TGR ELISA (K.R. Macaluso and A.L. Richards, unpub. data); thus, any soldier infected with R. felis would have reacted in the SFGR but not the TGR ELISA. The postdeployment seropositivity in US military personnel for antibodies against TGR, SFGR, and STGO at a titer ≥100 were 1.3% (117/9,249), 9.0% (805/8,918), and 0.5% (44/9,135), respectively (Figure). Seropositivity occurred for 10 (0.1%), 181 (2.0%), and 15 (0.2%) men who showed evidence of infection (seroconversion or 4-fold rise in antibody titer) with TGR, SFGR, and STGO, respectively, during their deployment to South Korea (Figure). The chance of a soldier having an infection with SFGR was significantly higher than the chance of having an infection with TGR or STGO (χ2 test, p<0.05) (analysis performed in SAS version 9.4; SAS Institute Inc., Cary, NC, USA). For personnel who seroconverted or had a 4-fold rise in titer to TGR, SFGR, or STGO, the age range was 19–49 (median 25) years, and job specialties were infantrymen (63.5%), fighting vehicle infantrymen (16.4%), indirect fire infantrymen (14.2%), and heavy anti-armor weapons infantrymen (5.9%). Figure Evidence of rickettsiosis or scrub typhus among US military personnel deployed to South Korea. Black bars indicate postdeployment serum samples from US military personnel with a titer ≥1:100 (seropositive) to typhus group rickettsiae (TGR), spotted ... These results indicate that many US military personnel were exposed to rickettsiae and orientiae before their deployment to South Korea (Figure), perhaps because of previous deployments around the world or because of exposure to rickettsial agents at home (8–10). However, 206 (2.2%) of the men became infected with either a typhus group (n = 10) or spotted fever group (n = 181) rickettsia or a scrub typhus group orientia (n = 15) during their deployment to South Korea. More SFGR infections occurred than TGR and STGO infections, although the pathogens for the latter infections (R. typhi and O. tsutsugamushi) are considered endemic to South Korea and are believed to affect the public and military health more than SFGR (3). The SFGR infections might correlate with recent observations of highly prevalent rickettsia-infected tick and R. felis–infected flea populations seen in South Korea (4,5). No evidence of co-infection was found in the men assessed during the deployment. These results suggest a risk for rickettsial disease, including scrub typhus and especially spotted fever, among US military personnel stationed in or visiting South Korea.

Outbreak of Cutaneous Leishmaniasis in Peruvian Military Personnel Undertaking Training Activities in the Amazon Basin, 2010

Military personnel deployed to the Amazon Basin are at high risk for cutaneous leishmaniasis (CL). We responded to an outbreak among Peruvian Army personnel returning from short-term training in the Amazon, conducting active case detection, lesion sample collection, and risk factor assessment. The attack rate was 25% (76/303); the incubation period was 2–36 weeks (median = 8). Most cases had one lesion (66%), primarily ulcerative (49%), and in the legs (57%). Real-time polymerase chain reaction (PCR) identified Leishmania (Viannia) braziliensis (59/61 = 97%) and L. (V.) guyanensis (2/61 = 3%). Being male (risk ratio [RR] = 4.01; P = 0.034), not wearing long-sleeve clothes (RR = 1.71; P = 0.005), and sleeping in open rooms (RR = 1.80; P = 0.009) were associated with CL. Sodium stibogluconate therapy had a 41% cure rate, less than previously reported in Peru (∼ 70%; P < 0.001). After emphasizing pre-deployment education and other basic prevention measures, trainees in the following year had lower incidence (1/278 = 0.4%; P < 0.001). Basic prevention can reduce CL risk in deployed militaries.

Population-based Neisseria gonorrhoeae, Chlamydia trachomatis and Trichomonas vaginalis prevalence using discarded, deidentified urine specimens previously collected for drug testing

We used a novel method to test for STIs among a non–healthcare-seeking military population in the San Diego region of California. Active-duty US Navy and Marine Corps personnel were randomly selected to provide urine specimens to Navy Drug Screening Laboratory, San Diego in October and November 2013 for the Department of Defense drug testing programme. If specimens screened negative for drugs (>99% of samples), urine specimens were discarded, deidentified and subsequently tested for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (GC) and Trichomonas vaginalis (TV) using the Aptima Combo 2 and TV assay as specified by the manufacturer (Hologic, San Diego, CA, USA). The Tigris direct tube sampling system was used for high-throughput nucleic acid amplification testing (NAAT). Urine specimens older than 6 days were not tested due to sample degradation concerns. The overall prevalence of CT was 2.1% (95% CI 1.79 …