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Didelphis marsupialis has been reported as a competent reservoir for trypanosomatid parasites infections. The aim of this study was to measure Trypanosoma cruzi, T. rangeli, and Leishmania spp. infection rates and to characterize discrete typing units (DTUs) of T. cruzi in D. marsupialis from two Chagas disease endemic sites in Panama. Blood from 57 wild-caught D. marsupialis were examined from two rural communities, Las Pavas (N = 18) and Trinidad de las Minas (N = 39). Twenty-two (38.60%) opossums were positive for flagellates by general hemoculture. T. cruzi infection was confirmed by positive hemoculture and/or kDNA based PCR performed in 31/57 (54.39%) blood samples from opossums. T. rangeli infection was confirmed by hemoculture and/or TrF/R2-Primer PCR assay applied on 12/57 (21.05%) blood samples. Nine (15.79%) D. marsupialis harbored T. cruzi/T. rangeli coinfections. All opossums tested negative for Leishmania spp. by PCR assays based on kDNA and HSP70 gene amplification. There was a significant association between T. cruzi infection and site (Fisher exact test, p = 0.02), with a higher proportion of T. cruzi infected opossums in Las Pavas (77.78%, n = 14/18) compared to Trinidad de las Minas (43.59%, n = 17/39). A significant association was found between habitat type and T. cruzi infection in opossums across both communities, (X 2  = 6.91, p = 0.01, df = 1), with a higher proportion of T. cruzi infection in opossums captured in forest remnants (76%, 19/25) compared to peridomestic areas (37.5%, 12/32). T. rangeli detection, but not T. cruzi detection, may be improved by culture followed by PCR. TcI was the only DTU detected in 22 T. cruzi samples using conventional and real-time PCR. Eight T. rangeli positive samples were characterized as KP1(-)/lineage C. Trypanosome infection data from this common synanthropic mammal provides important information for improved surveillance and management of Chagas disease in endemic regions of Panama., Competing Interests: The authors declare that there are no conflicts of interest., (© 2021 Published by Elsevier Ltd on behalf of Australian Society for Parasitology.)

Panama and all nations within the Mesoamerican region have committed to eliminate malaria within this decade. With more than 90% of the malaria cases in this region caused by Plasmodium vivax , an efficient national/regional elimination plan must include a comprehensive study of this parasite's genetic diversity. Here, we retrospectively analyzed P. vivax genetic diversity in autochthonous and imported field isolates collected in different endemic regions in Panama from 2007 to 2020, using highly polymorphic markers ( csp , msp -1, and msp -3α). We did the analysis using molecular techniques that are cost-effective for malaria molecular surveillance within Mesoamerica. Thus, we used molecular analyses that are feasible for malaria molecular surveillance within the region, and that can provide useful information for policy and decision making about malaria elimination. We also evaluated if haplotypes established by combining the genotypes found in these genes were associated with relevant epidemiological variables and showed structure across the transmission foci that have been observed in Panama. Ten different haplotypes were identified, some of them strongly associated with geographical origin, age, and collection year. Phylogenetic analysis of csp (central repeat domain) revealed that both major variant types (vk210 and vk247) were circulating in Panama. Variant vk247 was restricted to the eastern endemic regions, while vk210 was predominant (77.3%) and widespread, displaying higher diversity (14 alleles) and geographically biased alleles. The regional implications of these molecular findings for the control of P. vivax malaria to achieve elimination across Mesoamerica are discussed.

Background: More than 85% of the malaria cases in Panama occur in poor, rural and indigenous regions like Darien Province. Vector diversity, infection rate and spatial distribution are important entomological parameters of malaria transmission dynamics. Their understanding is crucial for the development of effective disease control strategies. The objective of this study was to determine the composition of Anopheles species, their natural infection rate and their geographic distribution to better understand the malaria transmission dynamics in Darién, Panama., Methods: Anophelines mosquitoes were captured during the rainy and dry season of 2016. We selected five communities where adult anophelines were collected using CDC light-traps, and through protective human-baited traps. Detection of natural infection and Plasmodium genotype were detected via nested PCR through the amplification of ssrRNA and the circumsporozoite protein gene (csp), respectively., Results: A total of 1,063 mosquitoes were collected mosquitoes were collected for the detection of natural infection with Plasmodium spp. Nine Anophelines species were identified, with the predominant species being: An. (Nys.) darlingi (45.0%) and An. (Nys.) albimanus (42.6%). Natural infection in An. (Nys.) albimanus with P. vivax was detected in one mosquito pool from the community Pueblo Tortuga (0.6%), three from Marraganti (1.7%), two from Bajo Chiquito (1.1%) and three pools from Alto Playona 3 (1.7%). For An. (Nys.) darlingi mosquitoes, we detected seven positive pools from the community Bajo Chiquito (4.0%), two pools from Marraganti (1.1%) and two pools from Alto Playona (1.1%). The P. vivax allelic variant VK210 was detected in infected mosquitoes., Conclusion: The results from this study provide new information on the transmission dynamics associated with anophelines vectors in the Darién region. This is the first report of natural P. vivax infection in An. (Nys.) darlingi and its incrimination as a potential malaria vector in this region of Panama. Additional studies are necessary to expand our knowledge and determine crucial parameters in malaria transmission in Darién, which in turn will aid the National Malaria Program in attaining an adequate malaria control strategy towards malaria elimination., Competing Interests: The authors declare that they have no competing interests. LC and JEC are members of the Sistema Nacional de Investigación (SNI), SENACYT, Panama. Any opinions, findings, conclusions or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture. The mention of firm names or trade products does not imply that they are endorsed or recommended by the U.S. Department of Agriculture over other firms or similar products not mentioned; the USDA is an equal opportunity employer. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Cutaneous Leishmaniasis transmission in the New World is observed in areas with rich sand fly species' faunas. The diversity and composition of sand fly species can change in response to seasonal weather and land use changes. Here, we present results from a two-year-long study where we collected, using Centers for Disease Control (CDC) light traps, sand flies from two rural areas, Las Pavas (LP) and Trinidad de las Minas (T) in western Panamá. Over 710 trap-nights, we collected 16,156 sand flies from 15 genera and 35 species. We identified 34 species in T, and the most abundant species collected was Nyssomyia trapidoi (Fairchild and Hertig, 1952) (n = 2278, 37%), followed by Psychodopygus panamensis (Shannon, 1926) (n = 1112, 18%), and Trichopygomyia triramula (Fairchild and Hertig, 1952) (n = 1063, 17%). In LP, we identified 26 species, and the most abundant species collected were Ty. triramula (n = 4729, 48%), and Ps. panamensis (n = 3444, 35%). We estimated a higher species' richness in T (Chao2 ± S.E.: 36.58 ± 3.84) than in LP (27.49 ± 2.28). In T, species' richness was significantly higher in the rainy season, but no seasonal differences were observed in LP. Species' assemblages were nested in the two areas. Phlebotomine sand fly species' abundance increased at the two sites during the rainy season. Our data suggest that seasonality is more important than land use as a factor driving sand fly species' diversity at the studied sites.

Panamá, together with all the nations in Mesoamerica, has committed to eliminate malaria from the region by 2020. As these countries approach malaria elimination and local transmission decreases, an active molecular surveillance to identify genotypes circulating along the border areas is particularly needed to accurately infer infection origin, drug resistance and disease propagation patterns in the region. This study evaluated the genetic diversity and allele frequencies of msp -1, msp -2 and glurp genes using different molecular analyses (nested PCR, PCR-restriction fragment length polymorphism (RFLP) and sequencing) from 106 autochthonous and imported P. falciparum isolates collected from different endemic areas in Panamá between 2003 and 2019. We also explored if P. falciparum genotypes assessed with these molecular markers were associated with relevant malaria epidemiological parameters using a multiple correspondence analysis. A strong association of certain local haplotypes with their geographic distribution in endemic areas, but also with parasite load and presence of gametocytes, was evidenced. Few multiclonal infections and low genetic diversity among locally transmitted P. falciparum samples were detected, consequent with the low transmission intensity of this parasite in Panamá, a pattern likely to be extended across Mesoamerica. In addition, several imported cases were genetically dissimilar to local infections and representative of more diverse extra-continental lineages.

Background: The present study provides a countrywide perspective of the malaria situation in Panamá over a long-term framework, with the purpose of identifying historical malaria resurgence events and their potential causes., Methods: A descriptive-ecological study was conducted by analysing demographic and epidemiological annual malaria time series data in Panamá (1884-2019) using several data sources. Malaria intensity indicators were calculated during the study period. The effects of El Niño Southern Oscillation on malaria transmission were also analysed using a retrospective analysis of malaria cases between 1957 and 2019., Results: Several factors were identified responsible for malaria resurgence in Panamá, mostly related with Malaria Control Programme weakening. During the past 20 years (2000-2019) malaria has progressively increased in prevalence within indigenous settlements, with a predominance of male cases and a high proportion (15% of total cases) in children less than 5 years old. During this period, a significant and increasing proportion of the Plasmodium falciparum cases were imported. Retrospective analysis (1957-2019) evidenced that ENSO had a significant impact on malaria transmission dynamics in Panamá., Conclusions: Data analysis confirmed that although authorities have been successful in focalizing malaria transmission in the country, there are still neglected issues to be solved and important intercultural barriers that need to be addressed in order to achieve elimination of the disease by 2022. This information will be useful for targeting strategies by the National Malaria Elimination Programme.

Synanthropic wild mammals can be important hosts for many vector-borne zoonotic pathogens. The aim of this study was determine the exposure of synanthropic mammals to two types of tick-borne pathogens in Panama, spotted fever group Rickettsia (SFGR) and Borrelia relapsing fever (RF) spirochetes. One hundred and thirty-one wild mammals were evaluated, including two gray foxes, two crab-eating foxes (from zoos), four coyotes, 62 opossum and 63 spiny rats captured close to rural towns. To evaluate exposure to SFGR, serum samples from the animals were tested by indirect immunofluorescence assay (IFA) using Rickettsia rickettsii and Candidatus Rickettsia amblyommii antigen. Immunoblotting was performed using Borrelia turicatae protein lysates and rGlpQ, to assess infection caused by RF spirochetes. One coyote (25%) and 27 (43%) opossums showed seroreactivity to SFGR. Of these opossums, 11 were seroreactive to C. R. amblyommii. Serological reactivity was not detected to B. turicatae in mammal samples. These findings may reflect a potential role of both mammals in the ecology of tick-borne pathogens in Panama., Competing Interests: The authors have declared that no competing interests exist.

Background: This study aims to describe the epidemiological and entomological factors associated with a recent malaria outbreak that occurred in 2012 in a socially marginalized population from Guna Yala Comarca in Panama., Methods: A descriptive and observational study was conducted by analysing demographic and epidemiological data from all malaria cases registered during 2012 in the Comarca Guna Yala, Panama. Malaria intensity indicators were calculated during the study period. Entomological evaluations were performed monthly, from October to December 2012, in the three communities that presented the most intense malaria transmission during the first semester of 2012. Anopheles breeding habitats were also characterized., Results: During the studied period, 6754 blood smears were examined (17.8 % of the total population), and 143 were confirmed as positive for Plasmodium vivax. A significant increase of malaria transmission risk indicators (API: 3.8/1000, SPR: 2.1 %) was observed in Guna Yula, when compared with previous years, and also in comparison with estimates from the whole country. Anopheles albimanus was the most abundant and widespread (877; 72.0 %) vector species found in the three localities, followed by Anopheles punctimacula (231; 19.0 %) and Anopheles aquasalis (110; 9.0 %). Three An. albimanus pools were positive for P. vivax, showing an overall pooled prevalence estimate of 0.014., Conclusions: Data analysis confirmed that during 2012 a malaria epidemic occurred in Guna Yala. Panama. This study provides baseline data on the local epidemiology of malaria in this vulnerable region of Panamá. This information will be useful for targeting control strategies by the National Malaria Control Programme.

Chagas disease, caused by Trypanosoma cruzi infection, is a zoonosis of humans, wild and domestic mammals, including dogs. In Panama, the main T. cruzi vector is Rhodnius pallescens, a triatomine bug whose main natural habitat is the royal palm, Attalea butyracea. In this paper, we present results from three T. cruzi serological tests (immunochromatographic dipstick, indirect immunofluorescence and ELISA) performed in 51 dogs from 24 houses in Trinidad de Las Minas, western Panama. We found that nine dogs were seropositive (17.6% prevalence). Dogs were 1.6 times more likely to become T. cruzi seropositive with each year of age and 11.6 times if royal palms where present in the peridomiciliary area of the dog's household or its two nearest neighbours. Mouse-baited-adhesive traps were employed to evaluate 12 peridomestic royal palms. All palms were found infested with R. pallescens with an average of 25.50 triatomines captured per palm. Of 35 adult bugs analysed, 88.6% showed protozoa flagellates in their intestinal contents. In addition, dogs were five times more likely to be infected by the presence of an additional domestic animal species in the dog's peridomiciliary environment. Our results suggest that interventions focused on royal palms might reduce the exposure to T. cruzi infection.

Background: Insecticide thermal fogging (ITF) is a tool to control vector borne diseases. Insecticide application success for vector control has been associated with housing materials and architecture. Vector abundance is correlated with weather changes. Nevertheless, housing quality and weather impacts on vector abundance have been unaccounted for in most New World insecticide control trials for leishmaniasis vectors., Methods: We conducted a 15 month insecticide control trial that included two deltamethrin [6 mg a.i.m-2] based ITF interventions in 12 of 24 monitored houses at Trinidad de Las Minas, a hyperendemic cutaneous leishmaniasis transmission village in western Panamá. During the study we followed sand fly (SF) abundance, keeping track of rainfall and quantified housing quality using an index based on architecture and construction materials., Results: We found a 50 to 80% reduction in SF density in the fogged houses when compared with control houses, while controlling for seasonal changes in SF abundance associated with rainfall. We found heterogeneities in the reductions, as abundance changed according to SF species: Lutzomyia gomezi, Lu. panamensis, Lu. dysponeta and Lu. triramula reduced in density between 40% and 90% after ITF. In contrast, Lu. trapidoi density increased 5% after ITF. Differences in the impact of ITF were associated with housing quality, the most destitute houses, i.e., those with features that ease insect entrance, had a disproportionally larger SF abundance, in some cases with increased domiciliary SF density following the ITF., Conclusion: Our results suggest the potential of insecticide application to control SF density and leishmaniasis transmission could depend on housing quality beyond insecticide efficiency.

American Cutaneous Leishmaniasis, ACL, is a zoonotic disease with a large richness of co-occurring vector species in transmission foci. Here, we describe changes in patterns of phlebotomine sand fly (Diptera: Psychodidae: Phlebotominae) species composition at the village of Trinidad de Las Minas, Capira, Panamá, a hyperendemic focus of ACL transmission, subjected to a vector control intervention with insecticide thermal fogging (ITF). Our study setting consisted of 24 houses, 12 subjected to two rounds of ITF and 12 kept as control. During 15 months (April 2010- June 2011) we monitored sand fly species composition and abundance with modified HP light traps inside (domicile) and outside (peridomicile) the studied houses. From 5628 sand flies collected, we were able to identify 5617 of the samples into 24 species, a number of species close to 25±1.6, the estimate from the Chao2 Index. The most abundant species were Lutzomya trapidoi (20%), Lu. gomezi (20%) and Lu. triramula (20%). Cluster analyses showed that most of the 24 houses had high similarity in relative abundance patterns of the six most common species, with only few peripheral houses not following the main cluster pattern. We also found that species richness was decreased to 22 species in the fogged houses, of which only 19 were found in the domiciliary environment. Changes in species richness were especially notorious at the end of the wet season. Our results suggest that species richness can decrease following ITF in domiciliary environments, primarily affecting the less common species.

We report a case of tracheal stenosis in a patient with immune thrombocytopenia who presented 4 yr after splenectomy. The 20-yr progression of the stenosis and management, including resection, is charted. The period after resection was complicated by wound infection, surgical emphysema, mediastinitis and dehiscence of the anastomosis of the trachea. The management of patients with tracheal lesions is discussed, but concentrates on airway care after tracheal resection when complications developed. A laryngeal mask airway was used to stabilize an uncuffed tracheal tube at the site of dehiscence.