Serologic Evidence of Scrub Typhus in the Peruvian Amazon.

Using a large, passive, febrile surveillance program in Iquitos, Peru, we retrospectively tested human blood specimens for scrub typhus group orientiae by ELISA, immunofluorescence assay, and PCR. Of 1,124 participants, 60 (5.3%) were seropositive, and 1 showed evidence of recent active infection. Our serologic data indicate that scrub typhus is present in the Peruvian Amazon.

BAES Scrotal flap: anatomical bases and usefulness in complex urethral reconstruction.

OBJECTIVE: To describe the anatomical characteristics and vascularization of the biaxial hair free scrotal flap (BAES-flap) and to detail its surgical application to reconstruction of the more complex urethral strictures. METHODS: We performed macro and micro anatomical dissections of the scrotum in 15 cryopreserved cadavers for the study of the arterial microvascularization of the BAES flap, and this anatomical knowledge has been implemented with the aim to improve the anterior and posterior urethra reconstructive surgical technique. For scrotal skin conditioning we performed definitive hair removal with the alexandrite laser. RESULTS: The BAES flap, thanks to its rich biaxial vascularization, its anatomical disposition over the urethral axis, and the suitable characteristics of hair free scrotal skin, has allowed us to perform successful one-step urethral reconstruction in complex cases such as panurethral disease, multioperated hypospadias, failed urethroplasties and obliterative stenosis. CONCLUSIONS: Detailed study of scrotal skin arterial vascularization is essential to design reliable and versatile genital skin flaps that result appropriate for the most complex reconstructive urethral surgery. The BAES scrotal flap complies with these requirements offering the patient a one step reconstructive option with a very satisfactory surgical experience over more than 20 years.

New Genetic Variants of Leptospira spp Characterized by MLST from Peruvian Isolates.

Leptospirosis is a zoonotic disease caused by the genus Leptospira, presenting complex and dynamic epidemiology. To determine the genetic variability and its phylogenetic relationship of Leptospira spp isolates from three sources in Iquitos (Peruvian Amazon) from 2002 to 2013, seven MLST genes were analyzed to obtain the Sequence Type (ST) and these sequences were concatenated for phylogenetic analysis. The genetic relationship between STs was determined with the goeBURST algorithm and genetic diversity was determined using DnaSP. Of 51 isolates, 48 were pathogenic belonging to five different species: Leptospira interrogans Nascimento 2004, Leptospira santarosai Feil 2004, Leptospira noguchii Haake 2021, Leptospira borgpetersenii Levett 2021, and Leptospira kirschneri Levett 2021. Of 20 STs identified, 60% corresponded to new genotypes circulating only in Peru. The genotypes ST17, ST37, and ST301 were recorded in rodents and humans. A high intraspecific genetic diversity was demonstrated in L. noguchi. The goeBURST analysis revealed three clonal complexes (CCs) and 16 singletons. The STs were found to show high genetic variability and phylogenetic and goeBURST analysis determined that the genotypes found did not form specific groups according to the source of infection or origin, which confirms the zoonotic potential of these STs in an area highly endemic for leptospirosis.

Venezuelan equine encephalitis and 2 human deaths, Peru.

Studies have suggested that enzootic strains of Venezuelan equine encephalitis (VEE) subtype ID in the Amazon region, Peru, may be less pathogenic to humans than are epizootic variants. Deaths of 2 persons with evidence of acute VEE virus infection indicate that fatal VEEV infection in Peru is likely. Cases may remain underreported.

Venezuelan equine encephalitis and upper gastrointestinal bleeding in child.

Venezuelan equine encephalitis (VEE) is reemerging in Peru. VEE virus subtype ID in Peru has not been previously associated with severe disease manifestations. In 2006, VEE virus subtype ID was isolated from a boy with severe febrile disease and gastrointestinal bleeding; the strain contained 2 mutations within the PE2 region.

Monitoring of leptospirosis seroprevalence in a colony of captive collared peccaries (Tayassu tajacu) from the Peruvian Amazon.

Leptospirosis, an endemic zoonoses, is maintained in the environment by several wildlife species in the Peruvian Amazon. In order to evaluate the possible role of collared peccaries (CP) in the maintenance this disease, two serological surveys of leptospirosis were performed and zootechnical parameters were monitored in a captive CP colony in an interval of 27 months. Total seroprevalence changed from 100% (n=27) to 86.4% (n=22), with reactions to a diversity of serogroups of zoonotic importance. Serological reactions to Leptospira licerasiae serogroup Iquitos, a new species recently identified locally and Leptospira interrogans serogroup Icterohaemorrhagiae were highly prevalent. The observation of leptospiral antibodies in both surveys, changes on serological reactions to different serogroups in large part of the herd and poor reproductive performances, provided an indication of the role of CP farms as a favourable environment for maintaining leptospirosis. Further research regarding the role of CP in the epidemiology of leptospirosis in the Peruvian Amazon is encouraged.

Rickettsial Disease in the Peruvian Amazon Basin.

Using a large, passive, clinic-based surveillance program in Iquitos, Peru, we characterized the prevalence of rickettsial infections among undifferentiated febrile cases and obtained evidence of pathogen transmission in potential domestic reservoir contacts and their ectoparasites. Blood specimens from humans and animals were assayed for spotted fever group rickettsiae (SFGR) and typhus group rickettsiae (TGR) by ELISA and/or PCR; ectoparasites were screened by PCR. Logistic regression was used to determine associations between patient history, demographic characteristics of participants and symptoms, clinical findings and outcome of rickettsial infection. Of the 2,054 enrolled participants, almost 2% showed evidence of seroconversion or a 4-fold rise in antibody titers specific for rickettsiae between acute and convalescent blood samples. Of 190 fleas (Ctenocephalides felis) and 60 ticks (Rhipicephalus sanguineus) tested, 185 (97.4%) and 3 (5%), respectively, were positive for Rickettsia spp. Candidatus Rickettsia asemboensis was identified in 100% and 33% of the fleas and ticks tested, respectively. Collectively, our serologic data indicates that human pathogenic SFGR are present in the Peruvian Amazon and pose a significant risk of infection to individuals exposed to wild, domestic and peri-domestic animals and their ectoparasites.

Clinical spectrum of pulmonary involvement in leptospirosis in a region of endemicity, with quantification of leptospiral burden.

BACKGROUND: Pulmonary involvement in leptospirosis remains poorly recognized in regions where it is endemic, despite reports of recent outbreaks and epidemic disease. METHODS: A prospective, population-based study was carried out to identify febrile patients exposed to Leptospira in urban and rural contexts in Iquitos, Peru. Evidence of exposure to Leptospira was obtained by serologic testing, and diagnosis of leptospirosis was confirmed in pulmonary cases by culture or quantitative real-time PCR assay. RESULTS: Of 633 consecutively enrolled febrile patients, 321 (50.7%) had antileptospiral IgM antibodies or high titers of antileptospiral antibodies. Seven patients with histories of only urban exposure to leptospires had severe pulmonary manifestations; of these, 5 patients died; 4 of the deaths were caused by pulmonary hemorrhage, and 1 was caused by acute respiratory distress syndrome and multiorgan failure. Real-time, quantitative PCR assay showed high levels of leptospiremia (>or=10(4) leptospires/mL) in most fatal cases; 1 patient, from whom tissue specimens were obtained at autopsy, had >or=10(5) leptospires/g of lung, kidney, and muscle tissue. DISCUSSION. This study demonstrates the underdiagnosis of leptospirosis in a region of high endemicity and the underrecognition of grave pulmonary complications. Pulmonary involvement in leptospirosis was present in urban but not rural areas. Presumptive treatment for leptospirosis should be initiated immediately in the appropriate epidemiological and clinical context.

Phylogenetic analysis of a novel molecular isolate of spotted fever group Rickettsiae from northern Peru: Candidatus Rickettsia andeanae.

Phylogenetic analysis of five rickettsial genes (17-kDa gene, gltA, ompB, ompA, and sca4) from two molecular isolates of Candidatus Rickettsia andeanae from two ticks (Amblyomma maculatum and Ixodes boliviensis) collected from two domestic horses living in two separate locations in northern Peru (Coletas and Naranjo) was conducted to more clearly characterize this recently reported novel spotted fever group (SFG) rickettsia. Following nested polymerase chain reaction (PCR) amplification of the 17-kDa gene, gltA, ompB, ompA, and sca4, amplicons were purified, sequenced, and compared to those downloaded from GenBank. Phylogenetic analyses of the Candidatus Rickettsia andeanae sequences generated from 17-kDa gene (483 bp), gltA (1185 bp), ompA (1598 bp), ompB (4839 bp), and sca4 (2634 bp) demonstrated that they aligned strongly with those of SFG rickettsiae. Moreover, the sequences of these five genes most closely aligned with the following rickettsiae: ompA: Rickettsia sp RpA4 (98.03%), R. sp DnS28 (97.90%), and R. rhipicephali and R. massiliae (97.11%); ompB: R. aeschlimannii (97.22%), R. rhipicephali (97.20%), and R. sp Bar 29 (97.10%); and sca4: R. massiliae (97.8%), R. rhipicephali, and R. slovaca (97.7%). These results from the additional phylogenetic analyses of Candidatus Rickettsia andeanae confirm its inclusion within, and distance and uniqueness from, other known SFG rickettsiae.

Evidence of rickettsial and leptospira infections in Andean northern Peru.

Between May and October 2002, a cluster of acute febrile illnesses occurred in the subtropical Andean foothills of Peru. Serologic evidence in villages where disease had been documented showed that the prevalence of IgM antibody to Leptospira ranged from 6% to 52%, that of IgM antibody to spotted fever group (SFG) rickettsia ranged from 10% to 19%, and that of IgM antibody to Coxiella burnetii from 1% to 15%. Measurement of IgG antibodies for SFG rickettsiae suggested that this disease was endemic. In contrast, IgG antibodies against C. burnetii were largely absent. In humans, microagglutination tests identified pathogenic variants of Leptospira. The presence of an SFG rickettsial infection was confirmed in four febrile patients following polymerase chain reaction and sequencing of the conserved 17-kD common antigen gene (htrA). Collectively, these analyses indicated that Rickettsia sp., C. burnetii, and Leptospira sp. were circulating in the region during the time of disease outbreak and implicate the involvement of an as yet undetermined SFG rickettsia in northwestern Peru.

A foodborne outbreak of brucellosis at a police station cafeteria, Lima, Peru.

Brucella melitensis is highly infectious for humans and can be transmitted to humans in a number of epidemiological contexts. Within the context of an ongoing brucellosis surveillance project, an outbreak at a Peruvian police officer cafeteria was discovered, which led to active surveillance (serology, blood culture) for additional cases among 49 police officers who had also eaten there. The cohort was followed up to 18 months regardless of treatment or symptoms. Active surveillance estimated the attack rate at 26.5% (13 of 49). Blood cultures from four cases were positive; these isolates were indistinguishable using multiple locus variable number tandem repeat analysis. This investigation indicates the importance of case tracking and active surveillance for brucellosis in the context of potential common source exposure. These results provide rationale for public health investigations of brucellosis index cases including the bioterrorism-related dissemination of Brucella.

[Genetic diversity of Peruvian isolates of Leptospira spp. through pulsed field gel electrophoresis]. / Diversidad genética de aislamientos peruanos de Leptospira spp. mediante electroforesis en gel de campo pulsado.

OBJECTIVES: Determine the genetic diversity of Peruvian isolations of Leptospira spp. through Pulsed Field Gel Electrophoresis (PFGE). MATERIALS AND METHODS: The PFGE methodology proposed by Galloway and Levett (2008) was standardized. A database including the PFGE profiles of 65 reference strains was prepared, and the technique was applied in 111 isolates of Leptospira spp. obtained in Peru between 2002 and 2010. RESULTS: A great generic diversity of serovars of circulating Leptospira spp. was determined in our country. 57 serovars were identified, 47 out of 97 pathogen isolates. Most frequent serovars were Icterohaemorrhagiae/Copenhageni (n=24) and Canicola (n=7). The most frequent species were L. santarosai (49,5%) and L. interrogans (37,1%). The distribution of species, clusters and serovars changed according to the source of isolate, the environmental context and the origin. CONCLUSIONS: There is great diversity of circulating serovars in Peru. There are genetic and epidemiological relations among isolates of different sources, and this is related to species, reservoir, environmental context and the origin of the isolate.

Iquitos virus: a novel reassortant Orthobunyavirus associated with human illness in Peru.

Oropouche (ORO) virus, a member of the Simbu serogroup, is one of the few human pathogens in the Orthobunyavirus genus in the family Bunyaviridae. Genetic analyses of ORO-like strains from Iquitos, Peru, identified a novel reassortant containing the S and L segments of ORO virus and the M segment of a novel Simbu serogroup virus. This new pathogen, which we named Iquitos (IQT) virus, was first isolated during 1999 from a febrile patient in Iquitos, an Amazonian city in Peru. Subsequently, the virus was identified as the cause of outbreaks of "Oropouche fever" during 2005 and 2006 in Iquitos. In addition to the identification of 17 isolates of IQT virus between 1999 and 2006, surveys for neutralizing antibody among Iquitos residents revealed prevalence rates of 14.9% for ORO virus and 15.4% for IQT virus. Limited studies indicate that prior infection with ORO virus does not seem to protect against disease caused with the IQT virus infection. Identification of a new Orthobunyavirus human pathogen in the Amazon region of Peru highlights the need for strengthening surveillance activities and laboratory capabilities, and investigating the emergence of new pathogens in tropical regions of South America.

Asymptomatic renal colonization of humans in the peruvian Amazon by Leptospira.

BACKGROUND: Renal carriage and shedding of leptospires is characteristic of carrier or maintenance animal hosts. Sporadic reports indicate that after infection, humans may excrete leptospires for extended periods. We hypothesized that, like mammalian reservoir hosts, humans develop asymptomatic leptospiruria in settings of high disease transmission such as the Peruvian Amazon. METHODOLOGY/PRINCIPAL FINDINGS: Using a cross-sectional study design, we used a combination of epidemiological data, serology and molecular detection of the leptospiral 16S rRNA gene to identify asymptomatic urinary shedders of Leptospira. Approximately one-third of the 314 asymptomatic participants had circulating anti-leptospiral antibodies. Among enrolled participants, 189/314 (59%) had evidence of recent infection (microscopic agglutination test (MAT0 >or=1:800 or ELISA IgM-positive or both). The proportion of MAT-positive and high MAT-titer (>or=1:800) persons was higher in men than women (p = 0.006). Among these people, 13/314 (4.1%) had Leptospira DNA-positive urine samples. Of these, the 16S rRNA gene from 10 samples was able to be sequenced. The urine-derived species clustered within both pathogenic (n = 6) and intermediate clades of Leptospira (n = 4). All of the thirteen participants with leptospiral DNA in urine were women. The median age of the DNA-positive group was older compared to the negative group (p

[Primary pneumonic plague with nosocomial transmission in La Libertad, Peru 2010]. / Peste neumónica primaria con transmisión intrahospitalaria en La Libertad, Perú 2010.

UNLABELLED: Pneumonic plague is one of the clinical forms of plague, of low frequency and high mortality, transmitted by direct inhalation of Yersinia pestis coming from an animal or from person to person. OBJECTIVE: To describe the clinical and epidemiological characteristics of the cases of primary pneumonic plague in an outbreak in the north of Peru. MATERIALS AND METHODS: The clinical records of the confirmed cases of primary pneumonic plague presenting in an outbreak occurring in La Libertad, in July 2010, were reviewed, also the search and contact investigation was performed. RESULTS: The index case was identified, as well as three additional cases, out of these, two were nosocomial infections related to the index case. The initial clinical presentation was characterized by sudden onset of fever, chills, myalgia and chest pain, which in less than 24 hours evolved to hypotension and cyanosis. The initiation of specific treatment varied from 2 to 12 days, and cases with prompt initiation of treatment had a better clinical outcome. The lethality was 50% (2/4). CONCLUSION: Nosocomial transmission of pneumonic plague in Peru is evidenced, with severe clinical manifestations and high lethality.

Arboviral etiologies of acute febrile illnesses in Western South America, 2000-2007.

BACKGROUND: Arthropod-borne viruses (arboviruses) are among the most common agents of human febrile illness worldwide and the most important emerging pathogens, causing multiple notable epidemics of human disease over recent decades. Despite the public health relevance, little is know about the geographic distribution, relative impact, and risk factors for arbovirus infection in many regions of the world. Our objectives were to describe the arboviruses associated with acute undifferentiated febrile illness in participating clinics in four countries in South America and to provide detailed epidemiological analysis of arbovirus infection in Iquitos, Peru, where more extensive monitoring was conducted. METHODOLOGY/FINDINGS: A clinic-based syndromic surveillance system was implemented in 13 locations in Ecuador, Peru, Bolivia, and Paraguay. Serum samples and demographic information were collected from febrile participants reporting to local health clinics or hospitals. Acute-phase sera were tested for viral infection by immunofluorescence assay or RT-PCR, while acute- and convalescent-phase sera were tested for pathogen-specific IgM by ELISA. Between May 2000 and December 2007, 20,880 participants were included in the study, with evidence for recent arbovirus infection detected for 6,793 (32.5%). Dengue viruses (Flavivirus) were the most common arbovirus infections, totaling 26.0% of febrile episodes, with DENV-3 as the most common serotype. Alphavirus (Venezuelan equine encephalitis virus [VEEV] and Mayaro virus [MAYV]) and Orthobunyavirus (Oropouche virus [OROV], Group C viruses, and Guaroa virus) infections were both observed in approximately 3% of febrile episodes. In Iquitos, risk factors for VEEV and MAYV infection included being male and reporting to a rural (vs urban) clinic. In contrast, OROV infection was similar between sexes and type of clinic. CONCLUSIONS/SIGNIFICANCE: Our data provide a better understanding of the geographic range of arboviruses in South America and highlight the diversity of pathogens in circulation. These arboviruses are currently significant causes of human illness in endemic regions but also have potential for further expansion. Our data provide a basis for analyzing changes in their ecology and epidemiology.

Human leptospirosis caused by a new, antigenically unique Leptospira associated with a Rattus species reservoir in the Peruvian Amazon.

As part of a prospective study of leptospirosis and biodiversity of Leptospira in the Peruvian Amazon, a new Leptospira species was isolated from humans with acute febrile illness. Field trapping identified this leptospire in peridomestic rats (Rattus norvegicus, six isolates; R. rattus, two isolates) obtained in urban, peri-urban, and rural areas of the Iquitos region. Novelty of this species was proven by serological typing, 16S ribosomal RNA gene sequencing, pulsed-field gel electrophoresis, and DNA-DNA hybridization analysis. We have named this species "Leptospira licerasiae" serovar Varillal, and have determined that it is phylogenetically related to, but genetically distinct from, other intermediate Leptospira such as L. fainei and L. inadai. The type strain is serovar Varillal strain VAR 010(T), which has been deposited into internationally accessible culture collections. By microscopic agglutination test, "Leptospira licerasiae" serovar Varillal was antigenically distinct from all known serogroups of Leptospira except for low level cross-reaction with rabbit anti-L. fainei serovar Hurstbridge at a titer of 1:100. LipL32, although not detectable by PCR, was detectable in "Leptospira licerasiae" serovar Varillal by both Southern blot hybridization and Western immunoblot, although on immunoblot, the predicted protein was significantly smaller (27 kDa) than that of L. interrogans and L. kirschneri (32 kDa). Isolation was rare from humans (2/45 Leptospira isolates from 881 febrile patients sampled), but high titers of MAT antibodies against "Leptospira licerasiae" serovar Varillal were common (30%) among patients fulfilling serological criteria for acute leptospirosis in the Iquitos region, and uncommon (7%) elsewhere in Peru. This new leptospiral species reflects Amazonian biodiversity and has evolved to become an important cause of leptospirosis in the Peruvian Amazon.

Diversidad genética de aislamientos peruanos de Leptospira spp. mediante electroforesis en gel de campo pulsado / Genetic diversity of Peruvian isolates of Leptospira spp. through pulsed field gel electrophoresis

Objetivos. Determinar la diversidad genética de aislamientos peruanos de Leptospira spp. mediante electroforesis en gel de campo pulsado (PFGE). Materiales y métodos. Se estandarizó la metodología de PFGE propuesta por Galloway y Levett (2008). Se elaboró una base de datos con los perfiles de PFGE de 65 cepas de referencia y se aplicó la técnica en 111 aislamientos de Leptospira spp. obtenidos en Perú entre 2002 y 2010. Resultados. Se determinó gran diversidad genética de serovares de Leptospira spp. circulantes en nuestro país. Se identificaron 57 serovares, 47 en 97 aislamientos patógenos. Los serovares más frecuentes fueron Icterohaemorrhagiae/Copenhageni (n=24) y Canicola (n=7). Las especies más frecuentes fueron L. santarosai (49,5%) y L. interrogans (37,1%). La distribución de especies, clusters y serovares varió según la fuente del aislamiento, el contexto ambiental y la procedencia. Conclusiones. Existe gran diversidad de serovares circulantes en el Perú, la cual está relacionada a la especie, el reservorio, el contexto ambiental y la procedencia del aislamiento. Se evidencia las relaciones genéticas y epidemiológicas entre aislamientos de diferentes fuentes, lo cual está relacionada a la especie, el reservorio, el contexto ambiental y la procedencia del aislamiento.

Objectives. Determine the genetic diversity of Peruvian isolations of Leptospira spp. through Pulsed Field Gel Electrophoresis (PFGE). Materials and methods. The PFGE methodology proposed by Galloway and Levett (2008) was standardized. A database including the PFGE profiles of 65 reference strains was prepared, and the technique was applied in 111 isolates of Leptospira spp. obtained in Peru between 2002 and 2010. Results. A great generic diversity of serovars of circulating Leptospira spp. was determined in our country. 57 serovars were identified, 47 out of 97 pathogen isolates. Most frequent serovars were Icterohaemorrhagiae/Copenhageni (n=24) and Canicola (n=7). The most frequent species were L. santarosai (49,5%) and L. interrogans (37,1%). The distribution of species, clusters and serovars changed according to the source of isolate, the environmental context and the origin. Conclusions. There is great diversity of circulating serovars in Peru. There are genetic and epidemiological relations among isolates of different sources, and this is related to species, reservoir, environmental context and the origin of the isolate.