The American Journal of Tropical Medicine and Hygiene - Volume 70, Issue 5, 2004

In vitro drug sensitivity to chloroquine (CQ), mefloquine (MQ) and quinine was investigated in 60 culture-adapted Plasmodium falciparum isolates from malaria patients in Padrecocha, a village in the Amazonian Department of Loreto, Peru. All isolates showed resistance to CQ, decreased susceptibility to quinine, and sensitivity to MQ. These isolates were examined for mutations in the P. falciparum multidrug resistance 1 (pfmdr1) and chloroquine resistance transporter (pfcrt) genes previously linked to CQ resistance. The mutations N86Y and D1246Y, two of the five mutations commonly observed in the pfmdr1 gene of CQ-resistant clones, were not found. The pfcrt mutation K76T, associated with CQ resistance, was identified in all the isolates tested. Sequence analysis of codons 72–76 in the pfcrt gene showed the haplotypes SVMNT and CVMNT.

The efficacy of chloroquine (CQ) and sulfadoxine-pyrimethamine (SP) was evaluated in 89 subjects greater than one year of age with uncomplicated P. falciparum malaria in the East Nusatenggara Province of Indonesia. Fever clearance time was longer in the SP group than in the CQ group. However, parasite clearance time was extended in subjects who received CQ compared with those who received SP. Major adverse events were not observed in either group, and no hospitalizations were required during the study. Treatment failure rates at day 28 were 69% for CQ and 8.5% for SP. In both treatment groups, gametocytemia developed during the follow-up period, but was more pronounced in the SP group, peaking at 94% on day 7. Regardless of treatment group, children < 10 years of age had significantly higher treatment failure rates than subjects ≥10 years of age (relative risk = 2.49), suggesting that acquired immunity influenced treatment outcomes in the presence of parasite drug resistance. Although a highly effective alternative to CQ for clearing infection, SP treatment also presented some potential drawbacks (e.g., increased and persistent gametocytemia). Replacement of CQ with SP as a first-line therapy, either alone or in combination with CQ, in those areas of Indonesia with high levels of CQ resistance should significantly improve treatment outcomes, particularly in vulnerable populations lacking clinical immunity. More efficacious and rapidly acting asexual stage treatments are generally associated with increased gametocyte clearance and combination therapy in areas where drug resistance is high or emerging may provide an additional means for reducing transmission.

The influence of several factors on parasite growth and 50% inhibitory concentration (IC50) for chloroquine was assessed. Most isolates stored at 4°C up to 72 hours grew when they were subsequently cultivated. However, parasite viability sharply decreased from 24 hours, and the mean chloroquine IC50 decreased significantly (P < 0.05). There was no evidence for selection of pre-culture populations due to storage alone. The time point when 3H-hypoxanthine was added (0 versus 18 hours) had no effect on the IC50 during the 42-hour incubation, but was associated with a lower IC50 when 3H-hypoxanthine was added after the initial 42-hour incubation during the 72-hour incubation. An increase in 3H-hypoxanthine incorporation and chloroquine IC50 was observed as the hematocrit was increased from 1.0% to 2.5%. For the same isolates, chloroquine IC50 values were generally similar when the initial parasitemia was between 0.1% and 0.5% but increased at higher (>0.75%) parasitemias. Based on these results, we recommend immediate cultivation after blood collection, a 42-hour incubation period with the addition of 3H-hypoxanthine at the beginning of incubation, a 1.5% hematocrit, and an initial parasitemia 0.1-0.5%. Further studies on serum substitutes, gas mixture, and comparison of isotopic and non-isotopic assays are needed to establish a standardized in vitro assay protocol.

Plasmodium falciparum infection during pregnancy may cause placental malaria and subsequently low birth weight, primarily through the placental sequestration of infected red blood cells. Measuring the burden of malaria during pregnancy usually involves determining the prevalence of placental malaria infection through microscopic examination of placental blood films, a difficult and error-prone process. A number of rapid diagnostic tests (RDTs) for malaria have been developed, most of them immunochromatographic dipstick assays. However, none have been tested for the direct determination of malaria antigen in placental blood. We undertook an evaluation of the Malaria Rapid Test (MAKROmed®) in determining placental malaria infection. The prevalence of placental parasitemia was 22.6% by microscopy, 51.0% by a polymerase chain reaction (PCR), and 43.1% by RDT. When the PCR was used as the gold standard, RDTs had a sensitivity of 89% and a specificity of 76%. The MAKROmed RDT was highly sensitive in the detection of placental malaria, but had lower than expected specificity.

To relate stability of malaria transmission to biologic characteristics of vector mosquitoes throughout the world, we derived an index representing the contribution of regionally dominant vector mosquitoes to the force of transmission. This construct incorporated published estimates describing the proportion of blood meals taken from human hosts, daily survival of the vector, and duration of the transmission season and of extrinsic incubation. The result of the calculation was displayed globally on a 0.5° grid. We found that these biologic characteristics of diverse vector mosquitoes interact with climate to explain much of the regional variation in the intensity of transmission. Due to the superior capacity of many tropical mosquitoes as vectors of malaria, particularly those in sub-Saharan Africa, antimalaria interventions conducted in the tropics face greater challenges than were faced by formerly endemic nations in more temperate climes.

Malaria vaccine RTS,S combined with thrombospondin-related anonymous protein (TRAP) and formulated with AS02A (RTS,S+TRAP/AS02A) is safe and immunogenic in adult humans and rhesus monkeys (Macaca mulatta). Here, RTS,S+TRAP/AS02A was administered on a 0-, 1-, and 3-month schedule to three cohorts of infant monkeys, along with adult comparators. Cohort 1 evaluated 1/5, 1/2, and full adult doses, with the first dose administration at one month of age; cohort 2 monkeys received full adult doses, with the first dose administration at one versus three months of age; and, cohort 3 compared infants gestated in mothers with or without previous RTS,S/AS02A immunization. Immunization site reactogenicity was mild. Some infants, including the phosphate-buffered saline only recipient, developed transient iron-deficiency anemia, which is considered a result of repeated phlebotomies. All RTS,S+TRAP/AS02A regimens induced vigorous antibody responses that persisted through 12 weeks after the last vaccine dose. Modest lymphoproliferative and ELISPOT (interferon-γ and interleukin-5) responses, particularly to TRAP, approximated adult comparators. RTS,S+TRAP/AS02A was safe and well tolerated. Vigorous antibody production and modest, selective cell-mediated immune responses suggest that RTS,S+TRAP/AS02A may be immunogenic in human infants.

An increase in maternal mortality risk during peak malaria transmission in endemic countries is thought to implicate malaria in maternal deaths. The purpose of this study was to evaluate changes in risk and causes of maternal death in relation to the malaria season at the main referral hospital in The Gambia. During the malaria season, there was a 168% increase in the maternal mortality ratio (MMR), a three-fold increase in the proportion of deaths due to anemia, and an eight-fold increase in the anemia MMR. Apart from a 5.4-fold increase in eclampsia, there was no significant change in the contribution of other causes of death. It is estimated that malaria may account for up to 93 maternal deaths per 100,000 live births.

Chagas disease, a major public health problem in Latin America, is caused by the protozoan parasite Trypanosoma cruzi and transmitted by hematophageous insects from the Triatominae subfamily. Control of this disease is based on domestic vector control with insecticides and improvements in housing. As with other vector-borne diseases, the identification of areas of high risk of disease transmission is a major prerequisite for the planning and implementation of cost-effective control programs. In this study, we explored the relationship between Triatoma dimidiata geographic distribution and bioclimatic factors in the Yucatán peninsula in Mexico, using geographic information systems, and developed predictive models of T. dimidiata domestic abundance and of its infection rates by T. cruzi. These predictions were then used to build the first natural transmission risk map for Chagas disease in the Yucatán peninsula, a tool that should prove very valuable for the implementation of effective vector control programs in the region.

To assess the magnitude of the Buruli ulcer (BU) problem in Cameroon, we conducted a cross-sectional survey in the Nyong River basin and identified on clinical grounds a total of 436 cases of active or inactive BU (202 and 234, respectively). Swab specimens were taken from 162 active cases with ulcerative lesions and in 135 of these (83.3%) the clinical diagnosis was confirmed by the IS2404 polymerase chain reaction. Most lesions (93%) were located on the extremities, with lower limbs being twice as commonly involved as upper limbs. The age of patients with active BU ranged from 2 to 90 years with a median age of 14.5 years. Vaccination with bacilli Calmette-Guérin appeared to protect children against more severe forms of BU with multiple lesions. We conclude that in Cameroon BU is endemic, at least in the study area, and that a comprehensive control program for BU in Cameroon is urgently needed.

Genotypes of Mycobacterium tuberculosis causing disease were investigated in pulmonary tuberculosis patients admitted to two adjacent wards of a tuberculosis hospital in Delhi, India. Genetic markers, the insertion sequence IS6110, a direct repeat sequence, and a polymorphic GC-rich sequence supported the circumstantial epidemiologic link between eight strains of M. tuberculosis, suggesting their possible involvement in small-scale, interpersonal transmission of both drug-sensitive and drug-resistant tuberculosis. This is the first report of a suspected acquisition of M. tuberculosis among hospitalized patients in India. The use of multiple molecular typing markers and techniques unequivocally identified the exact clonality of strains isolated from the hospital. The result of this study emphasizes the need for more comprehensive investigation of high-risk situations for tuberculosis transmission and long-term follow-up analysis for identifying such instances of unsuspected transmission.

Eleven cases of tuberculous peritonitis (TBP) in hemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) patients at the Kaohsiung Veterans General Hospital in Kaohsiung, Taiwan between 1991 and 2000 were studied retrospectively (six cases in the HD group and five cases in the CAPD group) The diagnosis of TBP was established by either positive ascite tuberculosis (TB) culture or biopsy-proven chronic granulomatous inflammation. Fever and abdominal pain were the most common symptoms, while leukocytosis and unexplained hypercalcemia were the most common laboratory findings. Ascite analysis showed a lymphocyte predominance in all HD patients, but in only 40% of the CAPD patients. The mean duration of a diagnosis by ascite TB cultures was six weeks, while a diagnosis confirmed by laparascopic biopsy took one week. All four fatal cases were diagnosed by TB cultures. Laparoscopic biopsy provided a rapid diagnosis and resulted in low morbidity and mortality in our patients. Based on our review of all possible abstracts found in a Medline search from 1966 to 2002 using the keywords tuberculosis, peritonitis, uremia, and dialysis, this may be the first study of TBP in different dialysis patients.

A total of 451 stool specimens were collected from children less than five years of age with acute diarrhea from Ifakara, Tanzania and processed to detect bacterial enteropathogens, parasites, and rotaviruses. These specimens were divided into 348 from the dry season and 103 from the rainy season. Overall, diarrheogenic Escherichia coli (35.7%) were the predominant enteropathogens, with enterotoxigenic E. coli, enteroaggregative E. coli, and enteropathogenic E. coli being the most prevalent. Moreover, enteroaggregative E. coli (63% versus 35.5%; P < 0.05), Shigella spp. (24% versus 12%; P < 0.05), and rotavirus (23% versus 4%; P < 0.05) were more prevalent in the dry season than in the rainy season and enterotoxigenic E. coli (51.6% versus 20%; P < 0.05) and Giardia lamblia (14% versus 1%; P < 0.05) were more prevalent in the rainy season.

The aim of this study was to investigate the incidence, epidemiology, and clinical manifestations of Q fever among hospitalized children in Greece. During a two-year period, 1,200 children with various clinical manifestations were prospectively tested for Coxiella burnetii infection by indirect immunofluorescence. Acute Q fever was diagnosed in eight (0.67%) patients. No chronic case of infection was detected. Multivariate analysis showed that children 11–14 years old and children reporting consumption of cheese from rural areas were at increased risk for this illness. Clinical manifestations of acute Q fever were pneumonia (two patients), meningitis (two), prolonged fever (two), hepatitis (one), and hemolytic-uremic syndrome (one). Q fever accounted for 2.9% of the cases with prolonged fever, 1.2% of the cases of meningitis, and 0.5% of the cases of pneumonia. Fever and headache were the most common symptoms at presentation. Our study indicates that Q fever is a rare cause of hospitalization during childhood.

Individuals with frequent exposures to agents spread through the parenteral route show a high prevalence of Leishmania seropositivity in Spain. However, the frequency of positive polymerase chain reaction (PCR) results for Leishmania in blood in this setting remains unknown. In this study, L. infantum kinetoplast DNA (kDNA) was found in blood from 23 (24%) of 95 asymptomatic individuals with a serum Leishmania antibody titer ≥ 1:20 and in none of 44 seronegative individuals. The greater the antibody titer, the greater the proportion of PCR-positive samples, but 16 (20%) of 81 individuals with antibody titers ≤ 1:40 tested positive by PCR. Nine (37%) PCR-positive and 22 (19%) (P = 0.03) PCR-negative individuals showed a positive leishmanin skin test result. This results show that a remarkable proportion of asymptomatic Leishmania-seropositive individuals at risk for parenterally transmitted infections carry Leishmania kDNA in blood. This is more common in subjects with a high serum Leishmania antibody level and a positive leishmanin skin test result.

We describe antibody isotype patterns resulting from Schistosoma japonicum infection among 155 individuals 5–76 years old from a community in Leyte, The Philippines. Their exposure, infection, and reinfection status had been observed in a previously described water contact study used to categorically classify them as putative resistant or susceptible individuals. Antigens tested for specific antibody isotype responses were soluble worm antigen preparation (SWAP) and soluble egg antigen (SEA) and a panel of recombinant molecules. The study was aimed primarily at evaluating antigen-specific antibody responses and their potential in inducing protection among putative resistant individuals. Specific antibody responses suggestive of an involvement in protection were an IgE response to SWAP among females less than 20 years of age (5–19) and IgA responses to SWAP in the younger (5–19 years) age groups. Compatible with other studies on human schistosomes, IgM reactivities to SWAP and SEA in the 5–19-year-old age group predicted susceptibility as did IgG4 responses to recombinant paramyosin.

We conducted a cohort study on impact of effects of eight years of annual ivermectin mass treatment administered in eight villages in Imo State, Nigeria. Physical and visual acuity examinations carried out in 462 persons in 1995, prior to the launching of mass drug administration with ivermectin, were compared with re-examinations of 411 (89%) of these same individuals in 2002. We found that gross visual impairment decreased from 16% to 1%, nodule prevalence decreased from 59% to 18%, and papular dermatitis was reduced from 15% to 2%. No change was seen in leopard skin rates (14%). The only incident lesions were three subjects from a single community having the appearance of new nodules (e.g., nodules not identified in the 1995 examinations). Differences in community coverage did not appear to influence the benefit from treatment of individual residents.

From June 14 to August 30, 2001 and June 10 to August 22, 2002, the mosquitoes of Oberlin, Ohio were surveyed and tested for West Nile virus (WNV). Mosquitoes were trapped weekly using gravid traps and CO2-baited Centers for Disease Control light traps at seven sites in 2001 and eight sites in 2002 in woodlots within the city limits. A total of 12,151 mosquitoes, representing 14 species and one species group, were collected in gravid traps, with Culex pipiens/restuans being the most abundant, followed by Ochlerotatus triseriatus. In light traps, 12,510 mosquitoes were collected, with Aedes vexans being the most abundant, followed by Culex pipiens/restuans. All gravid trap collections were tested for WNV via reverse transcriptase-polymerase chain reaction. West Nile virus was first detected in July 2002. Of all species tested, the Cx. pipiens/restuans species group exhibited the highest minimum infection rate (MIR) and the greatest percentage of positive pools. Both the MIR and percent of positive pools increased significantly with the date, although abundance of Cx. pipiens/restuans in gravid trap collections did not.

West Nile virus (WNV) was first detected in North America in New York City in 1999 and rapidly moved westward. Understanding the mechanisms by which the amplification cycle is reinitiated each year increases our ability to predict epizootics and geographic expansion of the disease. Such understanding is enhanced by knowledge of the patterns of infection in the vertebrate reservoir hosts. Blue jays (Cyanocitta cristata) may serve as reservoir hosts for WNV. We examined the influence of age and date on the prevalence of WNV in jay carcasses in Ohio during May–August 2002. Percent of carcasses that were infected increased significantly with time from 3% in May to more than 90% by August. We found no difference in prevalence between juvenile (nestlings and fledglings) and adult jays early in the season, which contradicts the expected pattern if the majority of the adults sampled in 2002 had been exposed to the virus in 2001. Therefore, jays infected in 2001 were unlikely to have been important in initiating the 2002 virus cycle in Ohio.

The aim of this study was to determine the predominant hepatitis B virus (HBV) genotype in the Solomon Islands and determine if there is any racial correlation between genotype and hepatitis B e antigen (HBeAg) production in Japanese and Melanesian individuals. A total of 403 serum samples from 206 Melanesian HBV carriers in the Solomon Islands and 197 Japanese carriers from Fukuoka (n = 106) and Okinawa (n = 91) living in Japan in 2001 were tested. The HBV genotypes of 206 Melanesian subjects were 114 with genotype C (55.3%) and 92 with genotype D (44.7%). The HBV genotypes of 197 Japanese subjects were 74 with genotype B (37.6%) and 123 with genotype C (62.4%). The total HBeAg prevalence of subjects in Fukuoka (36.8%) was significantly higher than that of subjects in Okinawa (14.3%) (P < 0.0001) and subjects in the Solomon Islands (35.0%; P = 0.0014, by the Mantel-Haenszel test). The genotype C prevalences were significantly different, ranging from 24.2% in Okinawa, to 54.4% in the Solomon Islands, to 95.3% in Fukuoka (all P < 0.0001, by chi-square test). The prevalence of HBeAg positivity was significantly higher in Melanesian genotype C subjects (42.0%) than Melanesian genotype D subjects (26.6%) (P = 0.0310). Similarly, the prevalence of HBeAg positivity was significantly higher in Japanese genotype C subjects (36.6%) than Japanese genotype B subjects (9.5%) (P < 0.0001). These findings indicate that that HBV was of genotypes C and D in the Solomon Islands, and that the pathogenesis of HBV-infected patients is related to HBV genotype rather than race.

This study estimated the clinical effectiveness of five different antibiotic regimens (doxycycline, chloramphenicol, ciprofloxacin, doxycycline plus chloramphenicol, and doxycycline plus ciprofloxacin) administered for infection with Rickettsia typhi in terms of the duration of the fever. Eighty-seven patients with endemic typhus were hospitalized between 1993 and 1998 at the General Hospital of Chania in Chania, Crete, Greece. The mean time to defervescence was 2.9 days for doxycycline, 4.0 days for chloramphenicol, and 4.2 days for ciprofloxacin. In patients receiving combinations of doxycycline plus chloramphenicol and doxycycline plus ciprofloxacin, fever subsided in 3.4 and 4.0 days, respectively. The outcome was favorable in all patients, and no deaths or relapses were observed within two months.