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Background: Accuracy of molecular tools for the identification of parasites that cause human cutaneous leishmaniasis (CL) could largely depend on the sampling method. Non-invasive or less-invasive sampling methods such as filter paper imprints and cotton swabs are preferred over punch biopsies and lancet scrapings for detection methods of Leishmania based on polymerase chain reaction (PCR) because they are painless, simple, and inexpensive, and of benefit to military and civilian patients to ensure timely treatment. However, different types of samples can generate false negatives and there is a clear need to demonstrate which sample is more proper for molecular assays. Methodology: Here, we compared the sensitivity of molecular identification of different Leishmania (Viannia) species from Peru, using three types of sampling: punch biopsy, filter paper imprint and lancet scraping. Different composite reference standards and latent class models allowed to evaluate the accuracy of the molecular tools. Additionally, a quantitative PCR assessed variations in the results and parasite load in each type of sample. Principal findings: Different composite reference standards and latent class models determined higher sensitivity when lancet scrapings were used for sampling in the identification and determination of Leishmania (Viannia) species through PCR-based assays. This was consistent for genus identification through kinetoplastid DNA-PCR and for the determination of species using FRET probes-based Nested Real-Time PCR. Lack of species identification in some samples correlated with the low intensity of the PCR electrophoretic band, which reflects the low parasite load in samples. Conclusions: The type of clinical sample can directly influence the detection and identification of Leishmania (Viannia) species. Here, we demonstrated that lancet scraping samples consistently allowed the identification of more leishmaniasis cases compared to filter paper imprints or biopsies. This procedure is inexpensive, painless, and easy to implement at the point of care and avoids the need for anesthesia, surgery, and hospitalization and therefore could be used in resource limited settings for both military and civilian populations. Author summary: Human cutaneous leishmaniasis affects low-income populations living in places far from health services. The early sampling and detection of the parasite are necessary for timely treatment, however there are no uniform sampling criteria, thus the sensitivity of molecular tests may vary due to various factors such as the type of sample, the time of the disease and the parasite load in the lesion. In this study, we compared the performance of three sampling methods for molecular identification of the genus and Leishmania (Viannia) species in Peru. Several analytical methods, including composite reference standards and latent class models, suggested that lancet scraping might be the best approach for parasite genus detection by kDNA-PCR and for parasite species determination by FRET probes-based Nested Real-Time PCR. [ABSTRACT FROM AUTHOR]

Background: In Peru, the information regarding sand fly vectors of leishmaniasis and bartonellosis in the Amazon region is limited. In this study, we carried out sand fly collections in Peruvian lowland and highland jungle areas using different trap type configurations and screened them for Leishmania and Bartonella DNA. Methodology/Principal findings: Phlebotomine sand flies were collected in Peruvian Amazon jungle and inter Andean regions using CDC light trap, UV and color LED traps, Mosquito Magnet trap, BG Sentinel trap, and a Shannon trap placed outside the houses. Leishmania spp. screening was performed by kDNA PCR and confirmed by a nested cytochrome B gene (cytB) PCR. Bartonella spp. screening was performed by ITS PCR and confirmed by citrate synthase gene (gltA). The PCR amplicons were sequenced to identify Leishmania and Bartonella species. UV and Blue LED traps collected the highest average number of sand flies per hour in low jungle; UV, Mosquito Magnet and Shannon traps in high jungle; and Mosquito Magnet in inter Andean region. Leishmania guyanensis in Lutzomyia carrerai carrerai and L. naiffi in Lu. hirsuta hirsuta were identified based on cytB sequencing. Bartonella spp. related to Bartonella bacilliformis in Lu. whitmani, Lu. nevesi, Lu. hirsuta hirsuta and Lu. sherlocki, and a Bartonella sp. related to Candidatus B. rondoniensis in Lu. nevesi and Lu. maranonensis were identified based on gltA gene sequencing. Conclusions/Significance: UV, Blue LED, Mosquito Magnet and Shannon traps were more efficient than the BG-Sentinel, Green, and Red LED traps. This is the first report of L. naiffi and of two genotypes of Bartonella spp. related to B. bacilliformis and Candidatus B. rondoniensis infecting sand fly species from the Amazon region in Peru. Author summary: Leishmaniasis and human bartonellosis, two neglected tropical diseases, are endemic in Peruvian Andean regions. However, no human bartonellosis cases has been reported in the Amazon jungle where leishmaniasis has high prevalence. The improvement of sand fly trapping methods and identification of putative Leishmania and Bartonella vectors across Andean and Amazonian regions are important steps towards the design of better control strategies and reducing the risk of transmission in endemic areas. In this paper the authors explored the effectiveness of different trap types, based in LED colors and odor attractants, for sand fly collections in endemic areas of leishmaniasis and bartonellosis. Also, we reported for the first time the detection of Bartonella DNA in sand fly species from the Peruvian Amazon jungle, and the first record in Peru of L. (V.) naiffi infecting Lu. hirsuta hirsuta. The information provided by this manuscript will serve as a baseline for future surveillance and intervention studies about sand fly species as potential vectors of Leishmania and Bartonella in highly endemic areas. [ABSTRACT FROM AUTHOR]

Background: Plasmodium vivax is the most predominant malaria species in Latin America, constituting 71.5% of malaria cases in 2021. With several countries aiming for malaria elimination, it is crucial to prioritize effectiveness of national control programs by optimizing the utilization of available resources and strategically implementing necessary changes. To support this, there is a need for innovative approaches such as genomic surveillance tools that can investigate changes in transmission intensity, imported cases and sources of reintroduction, and can detect molecular markers associated with drug resistance. Methodology/Principal findings: Here, we apply a modified highly-multiplexed deep sequencing assay: Pv AmpliSeq v2 Peru. The tool targets a newly developed 41-SNP Peru barcode for parasite population analysis within Peru, the 33-SNP vivaxGEN-geo panel for country-level classification, and 11 putative drug resistance genes. It was applied to 230 samples from the Peruvian Amazon (2007–2020), generating baseline surveillance data. We observed a heterogenous P. vivax population with high diversity and gene flow in peri-urban areas of Maynas province (Loreto region) with a temporal drift using all SNPs detected by the assay (nSNP = 2909). In comparison, in an indigenous isolated area, the parasite population was genetically differentiated (FST = 0.07–0.09) with moderate diversity and high relatedness between isolates in the community. In a remote border community, a clonal P. vivax cluster was identified, with distinct haplotypes in drug resistant genes and ama1, more similar to Brazilian isolates, likely representing an introduction of P. vivax from Brazil at that time. To test its applicability for Latin America, we evaluated the SNP Peru barcode in P. vivax genomes from the region and demonstrated the capacity to capture local population clustering at within-country level. Conclusions/Significance: Together this data shows that P. vivax transmission is heterogeneous in different settings within the Peruvian Amazon. Genetic analysis is a key component for regional malaria control, offering valuable insights that should be incorporated into routine surveillance. Author summary: Latin America is aiming towards malaria elimination. Genomic surveillance is crucial for a country's malaria strategy, aiding in understanding and stopping the spread of the disease. While widely used for another malaria species (Plasmodium falciparum), limited tools exist for tracking P. vivax, a significant player in malaria-endemic areas outside of Africa, and the primary cause of malaria in Latin America. In this study, we used a new tool, Pv AmpliSeq v2 Peru assay, to examine the genetic makeup of malaria parasites in the Peruvian Amazon. This tool helps us see how the parasites from different areas are connected and tracks markers that could indicate resistance to drugs. We found that the parasites from remote areas in the Amazon were genetically different from parasites in areas surrounding the main city of Iquitos, and parasites in a remote border community were genetically more similar to Brazilian parasites. We also show that the Pv AmpliSeq v2 Peru assay can be used to study parasites from other countries in Latin America, highlighting the broader application in the region. Considering that parasites are not constrained by borders and can easily spread between neighboring countries, a regional approach can be crucial for malaria elimination. [ABSTRACT FROM AUTHOR]

Background: Dengue fever is the most common arboviral disease in humans, with an estimated 50-100 million annual infections worldwide. Dengue fever cases have increased substantially in the past four decades, driven largely by anthropogenic factors including climate change. More than half the population of Peru is at risk of dengue infection and due to its geography, Peru is also particularly sensitive to the effects of El Niño Southern Oscillation (ENSO). Determining the effect of ENSO on the risk for dengue outbreaks is of particular public health relevance and may also be applicable to other Aedes-vectored viruses. Methods: We conducted a time-series analysis at the level of the district-month, using surveillance data collected from January 2000 to September 2018 from all districts with a mean elevation suitable to survival of the mosquito vector (<2,500m), and ENSO and weather data from publicly-available datasets maintained by national and international agencies. We took a Bayesian hierarchical modeling approach to address correlation in space, and B-splines with four knots per year to address correlation in time. We furthermore conducted subgroup analyses by season and natural region. Results: We detected a positive and significant effect of temperature (°C, RR 1.14, 95% CI 1.13, 1.15, adjusted for precipitation) and ENSO (ICEN index: RR 1.17, 95% CI 1.15, 1.20; ONI index: RR 1.04, 95% CI 1.02, 1.07) on outbreak risk, but no evidence of a strong effect for precipitation after adjustment for temperature. Both natural region and season were found to be significant effect modifiers of the ENSO-dengue effect, with the effect of ENSO being stronger in the summer and the Selva Alta and Costa regions, compared with winter and Selva Baja and Sierra regions. Conclusions: Our results provide strong evidence that temperature and ENSO have significant effects on dengue outbreaks in Peru, however these results interact with region and season, and are stronger for local ENSO impacts than remote ENSO impacts. These findings support optimization of a dengue early warning system based on local weather and climate monitoring, including where and when to deploy such a system and parameterization of ENSO events, and provide high-precision effect estimates for future climate and dengue modeling efforts. Author summary: The theoretical importance of the El Niño Southern Oscillation to infectious disease transmission is widely accepted, however few studies have quantified this effect or its interaction with local environment. Using surveillance data on outbreaks of dengue fever in Peru, we have found evidence that El Niño events increase the risk of dengue outbreaks in the high jungle and coast, and decrease this risk in the low jungle. Our findings are likely generalizable to other viruses which are, like dengue virus, are transmitted by Aedes aegypti mosquitoes, including Zika, chikungunya, and yellow fever. As climate change is expected to increase the frequency of El Niño events, these results indicate that arbovirus outbreaks may also increase, and that El Niño events may be leveraged to predict them. [ABSTRACT FROM AUTHOR]

Arboviruses transmitted by Aedes aegypti (e.g., dengue, chikungunya, Zika) are of major public health concern on the arid coastal border of Ecuador and Peru. This high transit border is a critical disease surveillance site due to human movement-associated risk of transmission. Local level studies are thus integral to capturing the dynamics and distribution of vector populations and social-ecological drivers of risk, to inform targeted public health interventions. Our study examines factors associated with household-level Ae. aegypti presence in Huaquillas, Ecuador, while accounting for spatial and temporal effects. From January to May of 2017, adult mosquitoes were collected from a cohort of households (n = 63) in clusters (n = 10), across the city of Huaquillas, using aspirator backpacks. Household surveys describing housing conditions, demographics, economics, travel, disease prevention, and city services were conducted by local enumerators. This study was conducted during the normal arbovirus transmission season (January—May), but during an exceptionally dry year. Household level Ae. aegypti presence peaked in February, and counts were highest in weeks with high temperatures and a week after increased rainfall. Univariate analyses with proportional odds logistic regression were used to explore household social-ecological variables and female Ae. aegypti presence. We found that homes were more likely to have Ae. aegypti when households had interruptions in piped water service. Ae. aegypti presence was less likely in households with septic systems. Based on our findings, infrastructure access and seasonal climate are important considerations for vector control in this city, and even in dry years, the arid environment of Huaquillas supports Ae. aegypti breeding habitat. Author summary: Mosquito transmitted infectious diseases are a growing concern around the world. The yellow fever mosquito (Aedes aegypti) has been responsible for recent major outbreaks of disease, including dengue fever and Zika. This mosquito prefers to bite humans and lay its eggs in artificial containers such as water tanks and planters. This makes Ae. aegypti well suited to become established in growing urban areas. Controlling these mosquitoes has been an important way to reduce the risk of disease transmission. Studies that are undertaken to understand local factors that contribute to the continued survival of the mosquito can be used to inform control practices. We conducted a study in the largest Ecuadorian city on the border of Peru where we collected adult mosquitoes from houses and surveyed household members about their behaviors, perceptions, and housing infrastructure associated with the mosquito vector. Mosquitoes were most numerous in weeks with high temperatures and a week after increased rainfall. We found that houses that had unreliable water service were more likely have mosquitoes present, while houses that used septic systems were less likely to have mosquitoes present. [ABSTRACT FROM AUTHOR]

Vector-borne diseases remain a significant public health threat in many regions of the world. Traditional vector surveillance and control methods have relied on active and passive surveillance programs, which are often costly and time-consuming. New internet-based vector surveillance systems have shown promise in removing some of the cost and labor burden from health authorities. We developed and evaluated the effectiveness of a new internet-based surveillance system, "AlertaChirimacha", for detecting Triatoma infestans (known locally by its Quechua name, Chirimacha), the Chagas disease vector, in the city of Arequipa, Peru. In the first 26 months post-implementation, AlertaChirimacha received 206 reports of residents suspecting or fearing triatomines in their homes or neighborhoods, of which we confirmed, through pictures or inspections, 11 (5.3%) to be Triatoma infestans. After microscopic examination, none of the specimens collected were infected with Trypanosoma cruzi. AlertaChirimacha received 57% more confirmed reports than the traditional surveillance system and detected 10% more infested houses than active and passive surveillance approaches combined. Through in-depth interviews we evaluate the reach, bilateral engagement, and response promptness and efficiency of AlertaChirimacha. Our study highlights the potential of internet-based vector surveillance systems, such as AlertaChirimacha, to improve vector surveillance and control efforts in resource-limited settings. This approach could decrease the cost and time horizon for the elimination of vector-mediated Chagas disease in the region., Competing Interests: The authors declare that they do not have any identifiable financial, personal, or professional interests that could influenced the findings presented in this paper., (Copyright: © 2023 Tamayo et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)