Search

Determining effective sampling methods for mosquitoes are among the first objectives in elucidating transmission cycles of vector-borne zoonotic disease, as the effectiveness of sampling methods can differ based on species, location, and physiological state. The Spissipes section of the subgenus Melanoconion of Culex represents an understudied group of mosquitoes which transmit Venezuelan equine encephalitis viruses (VEEV) in the Western Hemisphere. The objective of this study was to determine effective collection methods that target both blood-engorged and non-engorged females of the Spissipes section of Culex subgenus Melanoconion to test the hypothesis that favorable trapping methods differ between species and by physiological status within a species. Mosquitoes were collected using two commercially available traps, (CDC-light trap and BG-Sentinel trap), two novel passive traps (a novel mosquito drift fence and pop-up resting shelters), and two novel aspirators, (a small-diameter aspirator and a large-diameter aspirator) in Darién, Panama, and Florida, USA. The total number of female mosquitoes collected for each species was compared using rarefaction curves and diversity metrics. We also compared the utility of each trap for collecting total females and blood-engorged females of four Spissipes section mosquito species in Florida and Darién. In Darién, it was found that both blood-engorged and unfed females of Cx. pedroi were most effectively collected using the mosquito drift fence at 57.6% and 61.7% respectively. In contrast, the most unfed Cx. spissipes were collected using the mosquito drift fence (40.7%) while blood-engorged females were collected effectively by pop-up resting shelters (42.3%). In Florida, the best sampling technique for the collection of blood-engorged Cx. panocossa was the large diameter aspirator at 41.9%, while the best trap for collecting Cx. cedecei was the pop-up resting shelter at 45.9%. For unfed female Spissipes section mosquitoes in Florida, the CDC light trap with CO2 collected 84.5% and 98.3% of Cx. cedecei and Cx. panocossa respectively in Florida. Rarefaction analysis, and both the Shannon and Simpsons diversity indices all demonstrated that the mosquito drift fence was capable of collecting the greatest diversity of mosquito species regardless of location. The finding that the proportions of unfed and blood-engorged mosquitoes collected by traps differed both among and between species has implications for how studies of VEEV vectors will be carried out in future investigations. In Florida a combination of pop-up resting shelters and use of a large-diameter aspirator would be optimal for the collection of both VEEV vectors for host-use studies. Results demonstrate that traps can be constructed from common materials to collect mosquitoes for VEEV vector studies and could be assessed for their utilization in vectors of other systems as well. Unfortunately, no single method was effective for capturing all species and physiological states, highlighting a particular need for assessing trap utility for target species of a study. Author summary Venezuelan equine encephalitis virus is a potentially deadly human pathogen that is transmitted by an understudied group of tropical mosquitoes (Spissipes section of the Culex subgenus Melanoconion). These mosquitoes reside in swamps and jungles, and are challenging to identify, so studying their biology and importance in transmitting VEEV has been neglected. To further our understanding of VEEV, we compared six novel and commercially available traps to determine which traps are best for capturing these species in Panama and Florida. We found that several different types of traps are effective for collecting blood-engorged females of different species of VEEV vectors, and that traps utilized for collecting unfed specimens are not necessarily the same traps one should use for collecting blood-engorged females of the same species. Results of this study will enable researchers to better capture these important disease vectors, particularly those that are blood-engorged, which will allow researchers to determine host associations necessary for understanding VEEV transmission. This information can be used to make decisions on controlling vector species.

Freshwater organisms are often sensitive to pesticides, but their sensitivity varies across different taxa and with pesticide type and action mode, as shown by multiple acute toxicity tests. Such variability hampers predictions about how freshwater ecosystems may be altered by pesticide toxicity, which is especially critical for understudied areas of the world such as the tropics. Furthermore, there is little information about the sensitivity of some organisms that are key components of stream food webs; this is the case of litter-feeding detritivorous invertebrates, which contribute to the fundamental process of litter decomposition. Here, we examined the sensitivity of three common detritivores [Anchytarsus sp. (Coleoptera: Ptilodactylidae), Hyalella sp. (Amphipoda: Hyalellidae) and Lepidostoma sp. (Trichoptera: Lepidostomatidae)] to three pesticides commonly used (the insecticides bifenthrin and chlorpyrifos and the fungicide chlorothalonil) using acute (48 or 96 h) toxicity tests. Our study demonstrates that common-use pesticides provoke the mortality of half their populations at concentrations of 0.04–2.7 μg L-1. We found that all species were sensitive to the three pesticides, with the highest sensitivity found for chlorpyrifos. Additionally, we used the approach of species sensitivity distributions (SSD) to compare our study species with Daphnia magna and other temperate and tropical invertebrates. We found that the study species were among the most sensitive species to chlorpyrifos and chlorothalonil. Our results suggest that tropical detritivores merit special attention in ecological risk assessment of pesticides and highlight the need for accurate ecotoxicological information from ecologically relevant species in the tropics.

Abstract Background In Panama, arboviroses such as dengue fever, and more recently chikungunya fever and Zika disease, are transmitted by Aedes aegypti and Aedes albopictus. Their control is based on the elimination of breeding sites and fogging with pyrethroid insecticides. However, one of the significant issues derived from the prolonged use of pyrethroid insecticide is the development of resistance mechanisms, such as knockdown resistance or kdr. The objective of this study was to evaluate the presence of kdr mutations in a partial region of the VGSC gene in samples of wild-caught Aedes mosquitoes from different locations of the Metropolitan Region of Panama. Results Based on the analysis of 194 sequences of the VGSC gene, two kdr mutations (Ile1011Met and Val1016Gly) were detected in a specimen of Ae. aegypti. The frequency of kdr mutations in the evaluated samples of Ae. aegypti was 0.01. Conclusions This study provides evidence for a low frequency of kdr mutations in Ae. aegypti populations in Panama. It is possible that these changes have no impact on vector control interventions. To our knowledge, we report, for the first time in America the Val1016Gly mutation documented in Asia. In general terms, this result is highly relevant to the Aedes Control Programme in Panama since it constitutes a feasible approach for the timely detection of resistance as well as for the development of strategies.

American Cutaneous Leishmaniasis (ACL) is a neglected vector-borne zoonosis that persists despite increasing socio-economic development and urbanization in Panamá. Here, we investigate the association between environmental changes and spatio-temporal ACL transmission in the Republic of Panamá (1980–2012). We employ a macroecological approach, where patterns of variation in ACL incidence at the spatially coarse-grained scale of health areas are studied considering factors linked to the ecology of ACL transmission. We specifically study impacts of climatic variability, measured by the different phases of El Niño Southern Oscillation (ENSO), within diverse ecosystems and sand fly (Diptera: Psychodidae) vector species, as well as heterogeneous local climatic patterns, deforestation, population growth rates, and changes in social marginalization. We found that over the study period, patterns of ACL incidence: (i) were asynchronous with clusters changing from east to west of the Panamá Canal, (ii) trends increased in the west, and decreased or remained nearly constant in the east, independent of human population growth, (iii) generally increased in years following El Niño, and (iv) decreased as forest cover increased. We found no significant association between changes in socio-economic indicators and ACL transmission. Regarding vector abundance and presence, we found that studies had been biased to locations east of the Panamá canal, and that, in general, the abundance of dominant vector species decreased during the cold phase of ENSO. Finally, our results indicate that a macroecological approach is useful to understand heterogeneities related to environmental change impacts on ACL transmission.

BACKGROUND:Chikungunya virus (CHIKV) typically causes explosive epidemics of fever, rash and polyarthralgia after its introduction into naïve populations. Since its introduction in Panama in May of 2014, few autochthonous cases have been reported; most of them were found within limited outbreaks in Panama City in 2014 and Puerto Obaldia town, near the Caribbean border with Colombia in 2015. In order to confirm that Panama had few CHIKV cases compared with neighboring countries, we perform an epidemiological analysis of chikungunya cases reported from May 2014 to July 2015. Moreover, to understand this paucity of confirmed CHIKV cases, a vectorial analysis in the counties where these cases were reported was performed. METHODS:Chikungunya cases were identified at medical centers and notified to health authorities. Sera samples were analyzed at Gorgas Memorial Institute for viral RNA and CHIKV-specific antibody detection. RESULTS:A total of 413 suspected cases of CHIKV infections were reported, with incidence rates of 0.5 and 0.7 per 100,000 inhabitants in 2014 and 2015, respectively. During this period, 38.6% of CHIKV cases were autochthonous with rash and polyarthralgia as predominant symptoms. CHIKV and DENV incidence ratios were 1:306 and 1:34, respectively. A phylogenetic analysis of E1/E2 genomic segment indicates that the outbreak strains belong to the Asian genotype and cluster together with CHIKV isolates from other American countries during the same period. Statistical analysis of the National Vector Control program at the district level shows low and medium vector infestation level for most of the counties with CHIKV cases. This index was lower than for neighboring countries. CONCLUSIONS:Previous training of clinical, laboratory and vector workers allowed a good caption and detection of the chikungunya cases and fast intervention. It is possible that low/medium vector infestation level could explain in part the paucity of chikungunya infections in Panama.

Abstract Background Lutzomyia gomezi (Nitzulescu, 1931) is one of the main Leishmania (Vianna) panamensis vectors in Panama, and despite its medical significance, there are no population genetic studies regarding this species. In this study, we used the sequences of the mitochondrial gene cytochrome b/start of NADH1 and the nuclear elongation gene α-1 in order to analyze genetic variation and phylogeographic structure of the Lu. gomezi populations. Methods A total of 86 Lu. gomezi individuals were captured in 38 locations where cutaneous leishmaniasis occurred. DNA was extracted with phenol/chloroform methods and amplification of genes was performed using PCR primers for mitochondrial and nuclear markers. Results We found a total of 37 and 26 haplotypes of mitochondrial and nuclear genes, high haplotype diversity (h) for all three populations were detected with both molecular markers. Nucleotide diversity (π) was estimated to be high for all three populations with the mitochondrial marker, which was opposite to the estimate with the nuclear marker. In the AMOVA Φst recorded moderate (mitochondrial) and small (nuclear) population structure with statistical significance among populations. The analysis of the fixation index (Fst) used to measure the differentiation of populations showed that with the exception of the population located in the region of Bocas del Toro, the other populations presented with minor genetic differentiation. The median-Joining network of the mitochondrial marker reveled three clusters and recorded four haplotypes exclusively of localities sampled from Western Panama, demonstrating strong divergence. We found demographic population expansion with Fu´s Fs neutrality test. In the analysis mismatch distribution was observed as a bimodal curve. Conclusion Lu. gomezi is a species with higher genetic pool or variability and mild population structure, due to possible capacity migration and local adaptation to environmental changes or colonization potential. Thus, knowledge of the genetic population and evolutionary history is useful to understand the implications of different population genetic structures for cutaneous leishmaniasis epidemiology.

Neurotropic arboviral infections are an important cause of encephalitis. A zoonotic, vector-borne alphavirus, Madariaga virus (MADV; formerly known as South American eastern equine encephalitis virus), caused its first documented human outbreak in 2010 in Darien, Panama, where the genetically similar Venezuelan equine encephalitis virus (VEEV) is endemic. We report the results of a seroprevalence survey of animals and humans, illustrating contrasting features of MADV and VEEV ecology and epidemiology.Small mammals were trapped in 42 sites in Darien, Panama, using Sherman traps, Tomahawk traps, and mist nets for bats. Blood was tested for the presence of neutralizing antibodies to MADV and VEEV. In addition, bird sera collected in 2007 in Chagres, Panama, were tested for MADV and VEEV neutralizing antibodies. Viremia was ascertained by RT-PCR. Human exposure to these two viruses was determined by IgG ELISA, followed by plaque reduction neutralization tests. To identify relevant risk factors for MADV or VEEV exposure, logistic regression analysis was performed, and the most parsimonious model was selected based on the Akaike information criterion.The animal survey yielded 32 bats (16 species), 556 rodents (12 species), and 20 opossums (4 species). The short-tailed cane mouse (Zygodontomys brevicauda) found abundantly in pasture and farms, had the highest MADV seroprevalence (8.3%). For VEEV, the shrub and forest-dwelling long-whiskered rice rat (Transandinomys bolivaris) had the highest seroprevalence (19.0%). Viremia was detected in one animal (Z. brevicauda). Of the 159 bird sera (50 species) tested, none were positive for either virus. In humans (n = 770), neutralizing antibodies to MADV and VEEV were present in 4.8% and 31.5%, respectively. MADV seropositivity was positively associated with cattle ranching, farming, and fishing. Having VEEV antibodies and shrubs near the house diminished risk. Age, forest work, farming and fishing were risk factors for VEEV, while having MADV antibodies, glazed windows, waste pick-up and piped water were protective.Our findings suggest that the short-tailed cane mouse and the long-whiskered rice rat serve as hosts for MADV and VEEV, respectively. The preferred habitat of these rodent species coincides with areas associated with human infection risk. Our findings also indicate that MADV emerged recently in humans, and that the transmission cycles of these two sympatric alphaviruses differ spatially and in host utilization.

In Panama, species of the genus Lutzomyia are vectors of American cutaneous leishmaniasis (ACL). There is no recent ecological information that may be used to develop tools for the control of this disease. Thus, the goal of this study was to determine the composition, distribution and diversity of Lutzomyia species that serve as vectors of ACL. Sandfly sampling was conducted in forests, fragmented forests and rural environments, in locations with records of ACL. Lutzomyia gomezi, Lutzomyia panamensis and Lutzomyia trapidoi were the most widely distributed and prevalent species. Analysis of each sampling point showed that the species abundance and diversity were greatest at points located in the fragmented forest landscape. However, when the samples were grouped according to the landscape characteristics of the locations, there was a greater diversity of species in the rural environment locations. The Kruskal Wallis analysis of species abundance found that Lu. gomezi and Lu. trapidoi were associated with fragmented environments, while Lu. panamensis, Lutzomyia olmeca bicolor and Lutzomyia ylephiletor were associated with forested environments. Therefore, we suggest that human activity influences the distribution, composition and diversity of the vector species responsible for leishmaniasis in Panama.

Lutzomyia longipalpis is the primary vector of the parasite responsible for visceral leishmaniasis in the Americas. In the present study, Lu. longipalpis was found in a domiciliary area in Limón, a district in Capira, a region in which cutaneous leishmaniasis is endemic in Panama. Previously, this species has been found in a humid forest in this same region. Finding Lu. longipalpis in domiciliary areas indicates that this species may be adapting to new habitats and that it may play a role in the transmission of leishmaniasis in Panama.

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.

Lutzomyia longipalpis is the main vector of an important neglected disease in America. The genetic diversity of this vector was estimated in the population collected in two geographical areas separated by up to 37 km. We analyzed the sequence CB3-PDR / N1 N-PDR of 22 individuals obtaining a parameter of: h = 0.43 and tt = 0.0017 (Bona), h = 0.89, tt = 0.004 (The Lemon) with a genetic differentiation of kst = 0.03; p> 0.05 between them. Eight haplotypes were detected, of which it was shared. A significant differentiation was detected between the Panama-Colombia (kst = 0.98), Panama-Costa Rica (kst = 0.98) and Panama-Brazil (kst = 0.72) populations under the isolation model. The genetic inferences of this population can complement the dispersion information and provide important clues to understand the ecology of Lutzomyia longipalpis in Panama. [ABSTRACT FROM AUTHOR]

BACKGROUND:Cutaneous Leishmaniasis (CL) is a neglected tropical vector-borne disease. Sand fly vectors (SF) and Leishmania spp parasites are sensitive to changes in weather conditions, rendering disease transmission susceptible to changes in local and global scale climatic patterns. Nevertheless, it is unclear how SF abundance is impacted by El Niño Southern Oscillation (ENSO) and how these changes might relate to changes in CL transmission. METHODOLOGY AND FINDINGS:We studied association patterns between monthly time series, from January 2000 to December 2010, of: CL cases, rainfall and temperature from Panamá, and an ENSO index. We employed autoregressive models and cross wavelet coherence, to quantify the seasonal and interannual impact of local climate and ENSO on CL dynamics. We employed Poisson Rate Generalized Linear Mixed Models to study SF abundance patterns across ENSO phases, seasons and eco-epidemiological settings, employing records from 640 night-trap sampling collections spanning 2000-2011. We found that ENSO, rainfall and temperature were associated with CL cycles at interannual scales, while seasonal patterns were mainly associated with rainfall and temperature. Sand fly (SF) vector abundance, on average, decreased during the hot and cold ENSO phases, when compared with the normal ENSO phase, yet variability in vector abundance was largest during the cold ENSO phase. Our results showed a three month lagged association between SF vector abundance and CL cases. CONCLUSION:Association patterns of CL with ENSO and local climatic factors in Panamá indicate that interannual CL cycles might be driven by ENSO, while the CL seasonality was mainly associated with temperature and rainfall variability. CL cases and SF abundance were associated in a fashion suggesting that sudden extraordinary changes in vector abundance might increase the potential for CL epidemic outbreaks, given that CL epidemics occur during the cold ENSO phase, a time when SF abundance shows its highest fluctuations.