Isolation of viruses from mosquitoes (Diptera: Culicidae) collected in the Amazon Basin region of Peru.

As part of a comprehensive study on the ecology of arthropod-borne viruses in the Amazon Basin region of Peru, we assayed 539,694 mosquitoes captured in Loreto Department, Peru, for arboviruses. Mosquitoes were captured either by dry ice-baited miniature light traps or with aspirators while mosquitoes were landing on human collectors, identified to species, and later tested on Vero cells for virus. In total, 164 virus isolations were made and included members of the Alphavirus (eastern equine encephalomyelitis, Trocara, Una, Venezuelan equine encephalomyelitis, and western equine encephalomyelitis viruses), Flavivirus (Ilheus and St. Louis encephalitis), and Orthobunyavirus (Caraparu, Itaqui, Mirim, Murutucu, and Wyeomyia viruses) genera. In addition, several viruses distinct from the above-mentioned genera were identified to the serogroup level. Eastern equine encephalomyelitis virus was associated primarily with Culex pedroi Sirivanakarn & Belkin, whereas Venezuelan equine encephalomyelitis virus was associated primarily with Culex gnomatos Sallum, Huchings & Ferreira. Most isolations of Ilheus virus were made from Psorophora ferox (Von Humboldt). Although species of the Culex subgenus Melanoconion accounted for only 45% of the mosquitoes collected, 85% of the virus isolations were made from this subgenus. Knowledge of the viruses that are being transmitted in the Amazon Basin region of Peru will enable the development of more effective diagnostic assays, more efficient and rapid diagnoses of clinical illnesses caused by these pathogens, risk analysis for military/civilian operations, and development of potential disease control measures.

Haiti: absence of dengue hemorrhagic fever despite hyperendemic dengue virus transmission.

In 1994-1996, 185 strains of dengue (DEN) virus types 1, 2, and 4 were recovered from febrile United States and other United Nations military personnel in Haiti. We wondered whether risk factors for dengue hemorrhagic fever (DHF) existed and, if so, were DHF cases occurring among Haitian children. Dengue transmission rates were studied in 210 school children (6-13 years old) resident in Carrefour Borough, Port-au-Prince, Haiti. When sera were tested for plaque-reduction neutralizing antibodies to DEN 1-4 viruses, nearly 85% had antibodies to two or more DEN serotypes. The annual transmission rate was estimated at 30%, a rate observed in countries endemic for DHE Haitian DEN 2 isolates were genotype I, which are repeatedly associated with DHF cases in Southeast Asia and American regions. Despite positive virologic pre-conditions, DHF cases were not recorded by experienced Port-au-Prince pediatricians. These observations, which are reminiscent of those in Africa, provide further evidence of a dengue resistance gene in black populations.

Allpahuayo virus: a newly recognized arenavirus (arenaviridae) from arboreal rice rats (oecomys bicolor and oecomys paricola) in northeastern peru.

Allpahuayo virus was initially isolated from arboreal rice rats (Oecomys bicolor and Oecomys paricola) collected during 1997 at the Allpahuayo Biological Station in northeastern Peru. Serological and genetic studies identified the virus as a new member of the Tacaribe complex of the genus Arenavirus. The small (S) segment of the Allpahuayo virus prototype strain CLHP-2098 (Accession No. AY012686) was sequenced, as well as that of sympatric isolate CLHP-2472 (Accession No. AY012687), from the same rodent species. The S segment was 3382 bases in length and phylogenetic analysis indicated that Allpahuayo is a sister virus to Pichinde in clade A. Two ambisense, nonoverlapping reading frames were identified, which result in two predicted gene products, a glycoprotein precursor (GPC) and a nucleocapsid protein (NP). A predicted stable single hairpin secondary structure was identified in the intergenic region between GPC and NP. Details of the genetic organization of Allpahuayo virus are discussed.

Trocara virus: a newly recognized Alphavirus (Togaviridae) isolated from mosquitoes in the Amazon Basin.

This report describes Trocara virus, a newly recognized member of the genus Alphavirus, that has been isolated from Aedes serratus mosquitoes collected at two widely separated sites in the Amazon Basin. Biological, antigenic and genetic characteristics of the new virus are given. Results of these studies indicate that Trocara virus is the first member of a newly discovered antigenic complex within the family Togaviridae genus Alphavirus. The public health and veterinary importance of Trocara virus is still unknown.

Epidemiological and ecological characteristics of past dengue virus infection in Santa Clara, Peru.

To determine risk factors associated with dengue (DEN) virus infection among residents of Santa Clara, Peru, a rural Amazonian village near Iquitos, a cross-sectional serological, epidemiological and environmental survey was conducted. Demographic, social and behavioural information was obtained by standardized questionnaire from 1225 Santa Clara residents (61.3%) aged 5 years or older. Additional data were obtained on the environmental variables and immature mosquito species and abundance surrounding each household (n = 248). Sera that had been collected previously by the Peruvian Ministry of Health from residents were tested by an enzyme-linked immunosorbent assay (ELISA) for DEN virus IgG antibody. Antibody identity was verified as DEN by plaque reduction neutralization test. Data on individuals were analysed by univariate and multivariable methods, and independent sample t-tests. Spatial clustering was evaluated by comparing distances among DEN positive households. Overall, antibody prevalence was 29.4 % and more than doubled from the youngest to the oldest age groups, but did not differ by sex. Curiously, length of residence in Santa Clara was negatively associated with DEN virus antibodies. More frequent travel to Iquitos was positively associated with seroprevalence. Residents who obtained water from a river source rather than a local well also had significantly higher antibody prevalence. None of the environmental variables measured at each household corresponded to the patterns of antibody distribution. Of the larval mosquitoes found around residences, all were determined to be species other than Aedes. No evidence of spatial autocorrelation among antibody-positive households was detected. These results strongly suggested that recent DEN virus transmission did not occur in the village and that most infections of residents of this rural village were acquired while visiting the city of Iquitos.

Vector competence of Peruvian mosquitoes (Diptera: Culicidae) for epizootic and enzootic strains of Venezuelan equine encephalomyelitis virus.

Mosquitoes collected in the Amazon Basin, near Iquitos, Peru, were evaluated for their susceptibility to epizootic (IAB and IC) and enzootic (ID and IE) strains of Venezuelan equine encephalomyelitis (VEE) virus. After feeding on hamsters with a viremia of approximately 10(8) plaque-forming units of virus per milliliter, Culex (Melanoconion) gnomatus Sallum, Huchings, & Ferreira, Culex (Melanoconion) vomerifer Komp, and Aedes fulvus (Wiedemann) were highly susceptible to infection with all four subtypes of VEE virus (infection rates > or = 87%). Likewise, Psorophora albigenu (Peryassu) and a combination of Mansonia indubitans Dyar & Shannon and Mansonia titillans (Walker) were moderately susceptible to all four strains of VEE virus (infection rates > or = 50%). Although Psorophora cingulata (Fabricius) and Coquillettidia venezuelensis (Theobald) were susceptible to infection with each of the VEE strains, these two species were not efficient transmitters of any of the VEE strains, even after intrathoracic inoculation, indicating the presence of a salivary gland barrier in these species. In contrast to the other species tested, both Culex (Melanoconion) pedroi Sirivanakarn & Belkin and Culex (Culex) coronator Dyar & Knab were nearly refractory to each of the strains of VEE virus tested. Although many of the mosquito species found in this region were competent laboratory vectors of VEE virus, additional studies on biting behavior, mosquito population densities, and vertebrate reservoir hosts of VEE virus are needed to incriminate the principal vector species.

Emerging genetic diversity of HIV-1 in South America.

OBJECTIVE: Genotype determination and risk group analysis of HIV-1 infected individuals in selected regions of South America. DESIGN: Cross-sectional convenience sampling of HIV-1-positive individuals in Peru, Ecuador, Uruguay and Paraguay from March, 1994 through September, 1998. METHODS: HIV-1-positive subjects were identified through the national AIDS surveillance program in each country. A standardized questionnaire was used to obtain demographic, clinical and risk factor data on each study subject. Viral DNA was extracted from participants' peripheral blood mononuclear cells either directly or after co-cultivation. A nested PCR was used to obtain selected fragments of the envelope genes for genotyping by the heteroduplex mobility assay (HMA). A 600 bp sequence encompassing the V3 loop was sequenced from a selection of 23 of these samples for phylogenetic analysis and confirmation of HMA genotype. RESULTS: Among the 257 successfully genotyped HIV-1-positive samples, genotype B was found in 98.3% (228/232) of those obtained from subjects in Peru, Ecuador, and Paraguay. In contrast, 56% (14/25) of the samples from Uruguay were genotype F, and the remainder were genotype B. Genotype F was detected for the first time in Peru (2/224) and Paraguay (1/4), and genotype A for the first time in Peru (1/224). Phylogenetic analysis confirmed the genotype identified by HMA in the 23 samples sequenced. There was no detectable genetic clustering of HIV-1 within the different high-risk groups or geographic locations. CONCLUSIONS: These findings verify and extend the presence of several different HIV-1 genotypes in South America.

Nucleotide sequences and phylogeny of the nucleocapsid gene of Oropouche virus.

The nucleotide sequence of the S RNA segment of the Oropouche (ORO) virus prototype strain TRVL 9760 was determined and found to be 754 nucleotides in length. In the virion-complementary orientation, the RNA contained two overlapping open reading frames of 693 and 273 nucleotides that were predicted to encode proteins of 231 and 91 amino acids, respectively. Subsequently, the nucleotide sequences of the nucleocapsid genes of 27 additional ORO virus strains, representing a 42 year interval and a wide geographical range in South America, were determined. Phylogenetic analyses revealed that all the ORO virus strains formed a monophyletic group that comprised three distinct lineages. Lineage I contained the prototype strain from Trinidad and most of the Brazilian strains, lineage II contained six Peruvian strains isolated between 1992 and 1998, and two strains from western Brazil isolated in 1991, while lineage III comprised four strains isolated in Panama during 1989.

An outbreak of fulminant hepatitis delta in the Waorani, an indigenous people of the Amazon basin of Ecuador.

An outbreak of delta hepatitis occurred during 1998 among the Waorani of the Amazon basin of Ecuador. Among 58 people identified with jaundice, 79% lived in four of 22 Waorani communities. Serum hepatitis B surface antigen (HBsAg) was found in the sera of 54% of the jaundiced persons, and 14% of asymptomatic persons. Ninety-five percent of 105 asymptomatic Waorani had hepatitis B core (HBc) IgG antibody, versus 98% of 51 with jaundice. These data confirm that hepatitis B virus (HBV) infection is highly endemic among the Waorani. Sixteen of 23 (70%) HBsAg carriers identified at the onset of the epidemic had serologic markers for hepatitis D virus (HDV) infection. All 16 were jaundiced, where as only two of seven (29%) with negative HDV serology were jaundiced (P = .0006). The delta cases clustered in families, 69% were children and most involved superinfection of people chronically infected with HBV. The data suggest that HDV spread rapidly by a horizontal mode of transmission other than by the sexual route.

Hepatitis C in Peru: risk factors for infection, potential iatrogenic transmission, and genotype distribution.

A large seroepidemiologic and genotyping study of hepatitis C virus (HCV) was conducted in Lima, Peru, during the periods of 1986 to 1993 (cohort A) and 1994 (cohort B). Anti-HCV seroprevalence rates were 15.6% (216 of 1,389) and 11.7% (168 of 1,438), respectively. Low rates were seen among volunteer blood donors (1.1% and 0.8%). Anti-HCV rates were much higher among patients undergoing hemodialysis (43.7% and 59.3%), hemophiliacs (60.0% and 83.3%), in those more than 39 years old (18.2% and 26.0%), in females (25.0% and 27.4%), and in less-educated persons (16.9%). Age- and gender-adjusted risk factors in cohort B included blood transfusion history (adjusted odds ratio [AOR] = 29.8), prior organ transplantation (AOR = 9.1) or a history of hepatitis (AOR = 4.9), previous hospitalization (AOR = 3.7), a history of intravenous drug use (AOR = 3.5), prior major surgery (AOR = 2.6), a history of acupuncture (AOR = 2.1), previous dental procedures (AOR = 1.2), and prior medical injections (AOR = 1.04). The most prevalent HCV genotype was type 1 (86%), followed by type 3 (10%) and type 2 (2%). Transmission through unsafe injection-related and medical/dental procedures appears to play an important role in HCV infection among Peruvians.

Detection of pathogenic Leptospira spp. infections among mammals captured in the Peruvian Amazon basin region.

To identify potential zoonotic reservoirs of pathogenic leptospires in the Peruvian Amazon basin, wild mammals were trapped from July 1997 to December 1998 near the city of Iquitos. After extraction of nucleic acids from animal kidneys, DNA of pathogenic leptospires was identified by polymerase chain reaction (PCR) assays using one of two primer sets, one amplifying a region of the 23S rRNA gene, and the other amplifying a gene fragment specific for Leptospira spp (G1/G2 primers). Overall, 29% (40 of 136) of the mammals tested showed evidence of renal infection by Leptospira spp., including 20% (13 of 64) of the rodents, 39% (20 of 51) of the marsupials, and 35% (7 of 20) of the chiropterans (bats). Marsupials and chiropterans were implicated as more significant reservoir hosts of leptospires pathogenic to humans than previously recognized.

Failure of secondary infection with American genotype dengue 2 to cause dengue haemorrhagic fever.

BACKGROUND: Population-based epidemiological studies have shown that infection with dengue type 2 (DEN-2) virus in individuals previously infected with a different serotype of the virus is a major risk factor for dengue haemorrhagic fever and dengue shock syndrome. However, the western hemisphere was spared epidemics of these two syndromes, until the introduction of a southeast Asian DEN-2 genotype. Possibly American DEN-2 genotype strains lacked properties necessary to cause severe disease. We report on a major epidemic of DEN-2 in Peru in 1995, about 5 years after an epidemic of DEN-1 in the same population. METHODS: In Iquitos, a city of 344,686 inhabitants in Peru, cases of dengue fever were studied prospectively from 1990. Acute phase of illness serum samples from patients were tested for virus in C6/36 cells, and virus isolates were identified by immunofluorescence. Isolates of dengue 2 virus obtained from patients during an outbreak of mild febrile illness in 1995 were sequenced to determine the genotype. Serological analysis of paired samples from the patients was done with an IgM capture ELISA and an indirect IgG ELISA. In addition, serum samples collected annually between 1993 and 1996 from a large cohort of students were tested for dengue IgG antibody by an ELISA. Serum samples from a random sample of 129 students from this cohort were tested for dengue neutralising antibodies to quantify the serotype specific infection rates. FINDINGS: Among the 129 students (aged 7-20 years in 1993) who had serum samples available before and after the epidemic, 78 (60.5%) had a secondary DEN-2 virus infection. By extrapolation, 49,266 of the 81,479 children (aged 5-14 years) in Iquitos would have experienced such infections. From previous studies, between 887 and 10,247 cases of dengue haemorrhagic fever and dengue shock syndrome would have been expected. No cases were found. DEN-2 isolates were of the American genotype. INTERPRETATION: This prospective study shows that secondary infection by the American DEN-2 genotype did not cause dengue haemorrhagic fever and dengue shock syndrome.

Isolation and genetic characterization of a hantavirus (Bunyaviridae: Hantavirus) from a rodent, Oligoryzomys microtis (Muridae), collected in northeastern Peru.

This paper describes the isolation and partial genetic characterization of a hantavirus from a pygmy rice rat, Oligoryzomys microtis, collected within the urban area of Iquitos, Loreto Department, Peru. The virus, designated HTN-007, exhibited the highest degree of genetic similarity to Rio Mamore virus, which was originally described from the same rodent species in eastern Bolivia. Comparison of small and medium segment nucleotide sequence data from HTN-007 and Rio Mamore virus revealed 87% and 85% sequence identity, respectively. Based on these analyses, HTN-007 appears to be a variant of Rio Mamore virus. As such, it represents the first successful isolation of Rio Mamore virus and the first evidence for the existence of a hantavirus in Peru. Serologic studies done by immunofluorescence on blood samples of 56 O. microtis trapped at the collection site indicated that 21.4% had antibodies to hantavirus. In view of the proximity of this rodent species to humans and the close phylogenetic relationship of Rio Mamore virus to hantaviruses that have been associated with human disease, Rio Mamore virus may be a hantavirus of some public health importance in tropical South America.

Sexual practices and prevalence of HIV, HTLV-I/II, and Treponema pallidum among clandestine female sex workers in Lima, Peru.

BACKGROUND AND OBJECTIVES: A survey was conducted to determine the sexual behavior practices and prevalence of HIV-1, HTLV-I/II, and T. pallidum infections among unlicensed female sex workers (FSWs) in Lima, Peru. GOAL OF THIS STUDY: To assess the role of unlicensed FSWs as a potential source of retroviral and T. pallidum infection. STUDY DESIGN: Female sex workers from 15 brothels were enrolled. Sera samples were obtained and tested for antibodies to HIV-1, HTLV-I, and Treponema pallidum. RESULTS: Of 158 FSWs studied, all were negative for HIV-1, 6 were positive for HTLV-I, and 5 were positive for Treponema pallidum. Of their male clients, 75% used condoms, whereas only 3% reported condom use with their steady partners. When condoms were always used by clients, the history of a sexually transmitted disease was significantly reduced (p < 0.01), and the prevalence of HTLV-I (p < 0.05) and syphilis was lower among these workers. CONCLUSION: Data suggested that the low rate of sexually transmitted diseases among FSWs reflected the high rate of condom use by their male clients.

PIP: A survey was conducted to determine the prevalence of HIV-I, human T cell leukemia virus I and II (HTLV-I/II), and Treponema pallidum infection and the associated risk factors for the transmission of these sexually transmitted diseases (STDs) among unlicensed female sex workers (FSWs) in Lima, Peru, to further define their role as a potential source of infection. Unlicensed FSWs from 15 brothels were enrolled in this study from March to June 1994. Serum samples were collected and tested for antibodies to HIV-I, HTLV-I, HTLV-II, and Treponema pallidum. Results revealed that of the 158 FSWs studied, all were negative for HIV-I; 6 were positive for HTLV-I, and 5 had T. pallidum antibodies. Of their male clients, 75% had used condoms for the past 6 months, whereas only 3% reported condom use with their steady partners. Among the workers who stated that condoms were always used, the frequency of a history of STDs, including genital ulcers and inguinal adenopathies, was lower compared to occasional users. Similarly, the prevalence of HTLV-I infection and syphilis was lower among these workers. In conclusion, the study results suggested that the low rate of STDs among FSWs reflected a high rate of condom use.

Mayaro virus disease: an emerging mosquito-borne zoonosis in tropical South America.

This report describes the clinical, laboratory, and epidemiological findings on 27 cases of Mayaro virus (MV) disease, an emerging mosquito-borne viral illness that is endemic in rural areas of tropical South America. MV disease is a nonfatal, dengue-like illness characterized by fever, chills, headache, eye pain, generalized myalgia, arthralgia, diarrhea, vomiting, and rash of 3-5 days' duration. Severe joint pain is a prominent feature of this illness; the arthralgia sometimes persists for months and can be quite incapacitating. Cases of two visitors from the United States, who developed MV disease during visits to eastern Peru, are reported. MV disease and dengue are difficult to differentiate clinically.

Strongyloides stercoralis hyperinfection associated with human T cell lymphotropic virus type-1 infection in Peru.

A study was conducted in Lima, Peru to determine if patients with Strongyloides hyperinfection had human T cell lymphotropic virus type-1 (HTLV-I) infection. The study included patients with Strongyloides hyperinfection and a control group consisted of sex- and age-matched asymptomatic healthy individuals whose stools were negative for Strongyloides. A third group included patients with intestinal strongyloidiasis. Sera from each study subject were tested for HTLV-1/2I by an ELISA and Western blot. The HLTV-1 infection rates (85.7%, 18 of 21) were significantly (P < 0.001) associated with Strongyloides hyperinfection compared with the control group (4.7%, 1 of 21). The HTLV-1 rate (10%, 6 of 62) for patients with intestinal strongyloidiasis was significantly (P < 0.001) lower than patients with Strongyloides hyperinfection, but did not differ significantly (P > 0.05) from the control group. The association of HTLV-1 infection was observed among 17 of 19 patients more than 20 years of age and one of two younger patients. None had HTLV-2 infection. In conclusion, Strongyloides hyperinfection among Peruvian patients was highly associated with HTLV-1 infection.

Epidemiology of endemic Oropouche virus transmission in upper Amazonian Peru.

A cross-sectional serosurvey of a rural community near Iquitos, Peru was conducted to determine Oropouche (ORO) virus antibody prevalence and risk factors for human infection. Venous blood samples, and demographic, social, and risk factor data were obtained from people age five years of age and older who lived in the village of Santa Clara on the Nanay River, a tributary of the Amazon River. Sera were tested for ORO viral antibody by an ELISA. The specificity of viral antibody reactivity was determined by a standard plaque-reduction neutralization test. Interview data were analyzed by univariate and multiple logistic regression to determine which variables were statistically associated with previous ORO viral infection, as indicated by the presence of IgG antibody. Final models were evaluated based on log-likelihood and Wald chi-square. Clustering of seropositive residents within houses was analyzed by the method of Walter. Among 1,227 persons sampled, 33.7% (n=414) were positive for ORO viral IgG antibody. Overall, antibody prevalence was similar for males (33.9%) and females (33.6%), and increased significantly with age for both sexes to include more than half of persons more than 25 years of age. The length of residence in the village was positively associated with serologic status; persons who had moved to the village within the past 15 years were less likely to be seropositive than life-long residents of the same age. Antibody prevalence among immigrants who had lived in Santa Clara more than 15 years was similar to that in life-long residents. The activity most predictive of previous ORO viral infection was travel to forest communities and travel to Iquitos. No evidence of spatial heterogeneity in ORO virus antibody distribution was observed. Results suggested that endemic transmission of ORO virus in this region has been ongoing during many decades, and that people are at considerable risk of infection.

Notes on the phlebotomine sand flies from the Peruvian southeast--I. Description of Lutzomyia (Helcocyrtomyia) adamsi n. sp. (Diptera: Psychodidae).

A new species of phlebotomine sand fly, Lutzomyia adamsi n. sp., is described and illustrated from specimens collected during August 1994, in Sandia, Department of Puno-Peru. According to the Oficina Nacional de Evaluacion de Recursos Naturales(ONERN 1976), this locality is situated in the life zone known as humid, mountain, low tropical forest (bh-MBT). Many areas in the northern part of Puno, mainly in the Inambari and Tambopata basins, are endemic to leishmaniasis. These areas are the continuation of others, largely known as "leishmaniasic" in the departments of Cusco and Madre de Dios. The morphological characteristics indicated that this species belongs to the genus Lutzomyia, subgenus Helcocyrtomyia Barretto, 1962.

Geographic distribution and clinical description of leishmaniasis cases in Peru.

Studies were conducted from 1986 through 1993 to further define the geographic distribution and relative importance of different species of Leishmania as a cause of leishmaniasis in Peru. Patients with a clinical diagnosis of cutaneous and/or mucosal or diffuse cutaneous leishmaniasis were enrolled at the Naval Medical Research Institute Detachment (NAMRID) Laboratory in Lima, the Tropical Disease Clinic at San Marcos University Daniel A. Carrión, the Central Military Hospital, and a Ministry of Health hospital in Cusco, Peru. Clinical features, lesion aspirates, and biopsy tissue were obtained from each patient. All specimens were collected and assayed separately, including multiple specimens from some of the same patients for Leishmania parasites by inoculating aliquots of either aspirates or biopsy tissue suspensions onto Senekji's blood agar medium. Stocks of Leishmania isolates were used to prepare promastigotes to produce extracts for identifying the Leishmania species by the cellulose acetate electrophoresis enzyme technique. A total of 351 isolates of Leishmania were obtained from 350 patients who were infected primarily in the low and high jungle of at least 15 different Departments of Peru. Of the 351 isolates, 79% were identified as L. (V.) braziliensis, 7% as L. (V.) guyanensis, 10% as L. (V.) peruviana, 2% as L. (V.) lainsoni, and 1.7% as L. (L.) amazonensis. The clinical form of disease varied depending on the species of Leishmania, with L. (V.) braziliensis being associated most frequently with cutaneous, mucosal ulcers and mixed cutaneous and mucosal disease, and L. (V) peruviana, L. (V.) guyanensis, L. (V.) lainsoni with cutaneous lesions. Leishmania (L.) amazonensis was isolated from six patients, three with cutaneous lesions, one with mucosal lesions, and two with diffuse cutaneous lesions. Among all of the leishmaniasis cases, males were affected more frequently, and cases occurred among patients less than 10 to more than 51 years of age. These data further defined the geographic distribution and the relative frequency of Leishmania species associated with different clinical forms of leishmaniasis in Peru.

Venezuelan equine encephalitis febrile cases among humans in the Peruvian Amazon River region.

A survey was conducted from October 1, 1993 to June 30, 1995 to determine the arboviral etiologies of febrile illnesses in the city of Iquitos in the Amazon River Basin of Peru. The study subjects were patients who were enrolled at medical care clinics or in their homes by Peruvian Ministry of Health (MOH) workers as part of the passive and active disease surveillance program of the MOH. The clinical criterion for enrollment was the diagnosis of a suspected viral-associated, acute, undifferentiated febrile illness of < or = 5 days duration. A total of 598 patients were enrolled in the study. Demographic information, medical history, clinical data, and blood samples were obtained from each patient. The more common clinical features were fever, headache, myalgia, arthralgia, retro-ocular pain, and chills. Sera were tested for virus by the newborn mouse and cell culture assays. Viral isolates were identified initially by immunofluorescence using polyclonal antibody. An ELISA using viral-specific monoclonal antibodies and nucleotide sequence analysis were used to determine the specific variety of the viruses. In addition, thin and thick blood smears were observed for malaria parasites. Venezuelan equine encephalitis (VEE) virus subtype I, variety ID virus was isolated from 10 cases, including three cases in October, November, and December 1993, five cases in January and February 1994, and two cases in June 1995. The ELISA for IgM and IgG antibody indicated that VEE virus was the cause of an additional four confirmed and four presumptive cases, including five from January through March 1994 and three in August 1994. Sixteen cases were positive for malaria. The 18 cases of VEE occurred among military recruits (n = 7), agriculture workers (n = 3), students (n = 3), and general laborers (n = 5). These data indicated that an enzootic strain of VEE virus was the cause of at least 3% (18 of 598) of the cases of febrile illnesses studied in the city of Iquitos in the Amazon Basin region of Peru.