Long-term trends and spatial patterns of West Nile Virus emergence in California, 2004-2021.

AIMS: West Nile Virus (WNV) has remained a persistent source of vector-borne disease risk in California since first being identified in the state in 2003. The geographic distribution of WNV activity is relatively widespread, but varies considerably across different regions within the state. Spatial variation in human WNV infection depends upon social-ecological factors that influence mosquito populations and virus transmission dynamics. Measuring changes in spatial patterns over time is necessary for uncovering the underlying regional drivers of disease risk. METHODS AND RESULTS: In this study, we utilized statewide surveillance data to quantify temporal changes and spatial patterns of WNV activity in California. We obtained annual WNV mosquito surveillance data from 2004 through 2021 from the California Arbovirus Surveillance Program. Geographic coordinates for mosquito pools were analysed using a suite of spatial statistics to identify and classify patterns in WNV activity over time. CONCLUSIONS: We detected clear patterns of non-random WNV risk during the study period, including emerging hot spots in the Central Valley and non-random periods of oscillating WNV risk in Southern and Northern California subregions. Our findings offer new insights into 18 years of spatio-temporal variation in WNV activity across California, which may be used for targeted surveillance efforts and public health interventions.

Susceptibilities of Ugandan Plasmodium falciparum Isolates to Proteasome Inhibitors.

The proteasome is a promising target for antimalarial chemotherapy. We assessed ex vivo susceptibilities of fresh Plasmodium falciparum isolates from eastern Uganda to seven proteasome inhibitors: two asparagine ethylenediamines, two macrocyclic peptides, and three peptide boronates; five had median IC50 values <100 nM. TDI8304, a macrocylic peptide lead compound with drug-like properties, had a median IC50 of 16 nM. Sequencing genes encoding the ß2 and ß5 catalytic proteasome subunits, the predicted targets of the inhibitors, and five additional proteasome subunits, identified two mutations in ß2 (I204T, S214F), three mutations in ß5 (V2I, A142S, D150E), and three mutations in other subunits. The ß2 S214F mutation was associated with decreased susceptibility to two peptide boronates, with IC50s of 181 nM and 2635 nM against mutant versus 62 nM and 477 nM against wild type parasites for MMV1579506 and MMV1794229, respectively, although significance could not be formally assessed due to the small number of mutant parasites with available data. The other ß2 and ß5 mutations and mutations in other subunits were not associated with susceptibility to tested compounds. Against culture-adapted Ugandan isolates, two asparagine ethylenediamines and the peptide proteasome inhibitors WLW-vinyl sulfone and WLL-vinyl sulfone (which were not studied ex vivo) demonstrated low nM activity, without decreased activity against ß2 S214F mutant parasites. Overall, proteasome inhibitors had potent activity against P. falciparum isolates circulating in Uganda, and genetic variation in proteasome targets was uncommon.

Impact of Short-Term Storage on Ex Vivo Antimalarial Susceptibilities of Fresh Ugandan Plasmodium falciparum Isolates.

We measured susceptibilities of Ugandan Plasmodium falciparum isolates assayed on the day of collection or after storage at 4°C. Samples were incubated with serial dilutions of 8 antimalarials, and susceptibilities were determined from 72-h growth inhibition assays. Storage was associated with decreased growth and lower 50% inhibitory concentration values, but differences between assays beginning on day 0 or after 1 or 2 days of storage were modest, indicating that short-term storage before drug susceptibility determination is feasible.

Quantifying Spillover of an Urban Invasive Vector of Plant Disease: Asian Citrus Psyllid (Diaphorina citri) in California Citrus.

Urban environments frequently play an important role in the initial stages of biological invasions, often serving as gateways for non-native species, which may propagate to nearby natural and agricultural ecosystems in the event of spillover. In California, citrus trees are a dominant ornamental and food plant in urban and peri-urban environments. We studied the invasion dynamics of the Asian citrus psyllid (Diaphorina citri), which became widespread in urban areas of southern California starting in 2008, to understand the factors driving its more recent invasion in commercial citrus groves. Using a multi-year monitoring database, we applied a suite of models to evaluate the rate at which groves accrued their first D. citri detection and the cumulative number of detections thereafter. Grove characteristics and landscape context proved to be important, with generally higher invasion rates and more cumulative detections in groves that were larger, had more edge, or had more perforated shapes, with greater urbanization intensity favoring more rapid invasion, but with inconsistent effects of distance to roads among models. Notably, distance to urban or other grove occurrences proved to be among the most important variables. During the early phase of D. citri invasion in the region, groves closer to urban occurrences were invaded more rapidly, whereas more recently, invasion rate depended primarily on proximity to grove occurrences. Yet, proximity to urban and grove occurrences contributed positively to cumulative D. citri detections, suggesting a continued influx from both sources. These results suggest that inherent features of agroecosystems and spatial coupling with urban ecosystems can be important, temporally dynamic, drivers of biological invasions. Further consideration of these issues may guide the development of strategic responses to D. citri's ongoing invasion.

Drug susceptibility of Plasmodium falciparum in eastern Uganda: a longitudinal phenotypic and genotypic study.

BACKGROUND: Treatment and control of malaria depends on artemisinin-based combination therapies (ACTs) and is challenged by drug resistance, but thus far resistance to artemisinins and partner drugs has primarily occurred in southeast Asia. The aim of this study was to characterise antimalarial drug susceptibility of Plasmodium falciparum isolates from Tororo and Busia districts in Uganda. METHODS: In this prospective longitudinal study, P falciparum isolates were collected from patients aged 6 months or older presenting at the Tororo District Hospital (Tororo district, a site with relatively low malaria incidence) or Masafu General Hospital (Busia district, a high-incidence site) in eastern Uganda with clinical symptoms of malaria, a positive Giemsa-stained blood film for P falciparum, and no signs of severe disease. Ex-vivo susceptibilities to ten antimalarial drugs were measured using a 72-h microplate growth inhibition assay with SYBR Green detection. Relevant P falciparum genetic polymorphisms were characterised by molecular methods. We compared results with those from earlier studies in this region and searched for associations between drug susceptibility and parasite genotypes. FINDINGS: From June 10, 2016, to July 29, 2019, 361 P falciparum isolates were collected in the Busia district and 79 in the Tororo district from 440 participants. Of 440 total isolates, 392 (89%) successfully grew in culture and showed excellent drug susceptibility for chloroquine (median half-maximal inhibitory concentration [IC50] 20·0 nM [IQR 12·0-26·0]), monodesethylamodiaquine (7·1 nM [4·3-8·9]), pyronaridine (1·1 nM [0·7-2·3]), piperaquine (5·6 nM [3·3-8·6]), ferroquine (1·8 nM [1·5-3·3]), AQ-13 (24·0 nM [17·0-32·0]), lumefantrine (5·1 nM [3·2-7·7]), mefloquine (9·5 nM [6·6-13·0]), dihydroartemisinin (1·5 nM [1·0-2·0]), and atovaquone (0·3 nM [0·2-0·4]). Compared with results from our study in 2010-13, significant improvements in susceptibility were seen for chloroquine (median IC50 288·0 nM [IQR 122·0-607·0]; p<0·0001), monodesethylamodiaquine (76·0 nM [44·0-137]; p<0·0001), and piperaquine (21·0 nM [7·6-43·0]; p<0·0001), a small but significant decrease in susceptibility was seen for lumefantrine (3·0 nM [1·1-7·6]; p<0·0001), and no change in susceptibility was seen with dihydroartemisinin (1·3 nM [0·8-2·5]; p=0·64). Chloroquine resistance (IC50>100 nM) was more common in isolates from the Tororo district (11 [15%] of 71), compared with those from the Busia district (12 [4%] of 320; p=0·0017). We showed significant increases between 2010-12 and 2016-19 in the prevalences of wild-type P falciparum multidrug resistance protein 1 (PfMDR1) Asn86Tyr from 60% (391 of 653) to 99% (418 of 422; p<0·0001), PfMDR1 Asp1246Tyr from 60% (390 of 650) to 90% (371 of 419; p<0·0001), and P falciparum chloroquine resistance transporter (PfCRT) Lys76Thr from 7% (44 of 675) to 87% (364 of 417; p<0·0001). INTERPRETATION: Our results show marked changes in P falciparum drug susceptibility phenotypes and genotypes in Uganda during the past decade. These results suggest that additional changes will be seen over time and continued surveillance of susceptibility to key ACT components is warranted. FUNDING: National Institutes of Health and Medicines for Malaria Venture.

Associations between Varied Susceptibilities to PfATP4 Inhibitors and Genotypes in Ugandan Plasmodium falciparum Isolates.

Among novel compounds under recent investigation as potential new antimalarial drugs are three independently developed inhibitors of the Plasmodium falciparum P-type ATPase (PfATP4): KAE609 (cipargamin), PA92, and SJ733. We assessed ex vivo susceptibilities to these compounds of 374 fresh P. falciparum isolates collected in Tororo and Busia districts, Uganda, from 2016 to 2019. Median IC50s were 65 nM for SJ733, 9.1 nM for PA92, and 0.5 nM for KAE609. Sequencing of pfatp4 for 218 of these isolates demonstrated many nonsynonymous single nucleotide polymorphisms; the most frequent mutations were G1128R (69% of isolates mixed or mutant), Q1081K/R (68%), G223S (25%), N1045K (16%), and D1116G/N/Y (16%). The G223S mutation was associated with decreased susceptibility to SJ733, PA92, and KAE609. The D1116G/N/Y mutations were associated with decreased susceptibility to SJ733, and the presence of mutations at both codons 223 and 1116 was associated with decreased susceptibility to PA92 and SJ733. In all of these cases, absolute differences in susceptibilities of wild-type (WT) and mutant parasites were modest. Analysis of clones separated from mixed field isolates consistently identified mutant clones as less susceptible than WT. Analysis of isolates from other sites demonstrated the presence of the G223S and D1116G/N/Y mutations across Uganda. Our results indicate that malaria parasites circulating in Uganda have a number of polymorphisms in PfATP4 and that modestly decreased susceptibility to PfATP4 inhibitors is associated with some mutations now present in Ugandan parasites.

Spatiotemporal trends of cutaneous leishmaniasis in Costa Rica.

BACKGROUND: Cutaneous leishmaniasis (CL) remains an important neglected tropical disease in Costa Rica, which has one of the largest burdens of this disease in Latin America. METHODS: We identified district-level hotspots of CL from 2006 to 2017 and conducted temporal analysis to identify where hotspots were increasing across the country. RESULTS: Clear patterns of CL risk were detected, with persistent hotspots located in the Caribbean region, where risk was also found to be increasing over time in some areas. CONCLUSIONS: We identify spatiotemporal hotspots, which may be used in support of the leishmaniasis plan of action for the Americas.

Lead Optimization of Second-Generation Acridones as Broad-Spectrum Antimalarials.

The global impact of malaria remains staggering despite extensive efforts to eradicate the disease. With increasing drug resistance and the absence of a clinically available vaccine, there is an urgent need for novel, affordable, and safe drugs for prevention and treatment of malaria. Previously, we described a novel antimalarial acridone chemotype that is potent against both blood-stage and liver-stage malaria parasites. Here, we describe an optimization process that has produced a second-generation acridone series with significant improvements in efficacy, metabolic stability, pharmacokinetics, and safety profiles. These findings highlight the therapeutic potential of dual-stage targeting acridones as novel drug candidates for further preclinical development.

Spatiotemporal dynamics of the Southern California Asian citrus psyllid (Diaphorina citri) invasion.

Biological invasions are governed by spatial processes that tend to be distributed in non-random ways across landscapes. Characterizing the spatial and temporal heterogeneities of the introduction, establishment, and spread of non-native insect species is a key aspect of effectively managing their geographic expansion. The Asian citrus psyllid (Diaphorina citri), a vector of the bacterium associated with huanglongbing (HLB), poses a serious threat to commercial and residential citrus trees. In 2008, D. citri first began expanding northward from Mexico into parts of Southern California. Using georeferenced D. citri occurrence data from 2008-2014, we sought to better understand the extent of the geographic expansion of this invasive vector species. Our objectives were to: 1) describe the spatial and temporal distribution of D. citri in Southern California, 2) identify the locations of statistically significant D. citri hotspots, and 3) quantify the dynamics of anisotropic spread. We found clear evidence that the spatial and temporal distribution of D. citri in Southern California is non-random. Further, we identified the existence of statistically significant hotspots of D. citri occurrence and described the anisotropic dispersion across the Southern California landscape. For example, the dominant hotspot surrounding Los Angeles showed rapid and strongly asymmetric spread to the south and east. Our study demonstrates the feasibility of quantitative invasive insect risk assessment with the application of a spatial epidemiology framework.

Knowledge and prevention of tick-borne diseases vary across an urban-to-rural human land-use gradient.

We sought to determine the behavioral risk of exposure to tick-borne diseases across a human land-use gradient in a region endemic for diseases transmitted by the lone star tick. We measured the knowledge, attitudes, and preventive behaviors of visitors to 14 suburban, exurban, and rural recreational parks. A structured interview was conducted to determine respondents' (n=238) knowledge of tick-borne disease risk, perceived susceptibility to tick-borne disease, and tick bite prevention behaviors. We found significant differences across park types for most personal protective behaviors. Individuals in exurban parks were more likely to perform frequent tick checks and use chemical insect repellents compared to other park types (p<0.001), while suburban park visitors were more likely to avoid tick habitats (p<0.05). Disparities exist in the level of knowledge, perceived personal risk, and use of preventive measures across the human land-use gradient, suggesting that targeted public health intervention programs could reduce behavioral exposure risk by addressing specific gaps in knowledge and prevention.

US university response to H1N1: a study of access to online preparedness and response information.

BACKGROUND: The recent outbreaks of severe acute respiratory syndrome, H5N1 (avian influenza), and, most recently, the novel H1N1 influenza pandemic of 2009 have raised awareness of the danger of new and emerging infections. Preparedness and response plans for such outbreaks are crucial, and given the centrality of the Internet as a source of information on university and college campuses, such plans should be made available at pandemic-dedicated university Web sites. The information on these sites must be comprehensive, accessible, and tailored to the specific circumstances of individual schools. METHODS: An Internet-based search was conducted in September 2009 to evaluate university Web sites for influenza-specific information in a sample of 51 universities. Web sites were assessed by applying a set of key words and a list of 10 indicators used as measures of accessibility and comprehensiveness. RESULTS: Of the 51 universities evaluated, only 9 (17.6%) either had no influenza Web site or had a university influenza preparedness plan with no dedicated Web site. Only 6 (14.3%) of the schools with influenza specific Web sites had information for parents, with 23 (54.8%) providing information specifically for faculty and staff, and 24 (57.1%) providing information specifically to students. CONCLUSION: We found no guidelines for maximizing the access to and effectiveness of online pandemic communications at institutions of higher learning. Until such time as appropriate guidelines are developed, university authorities must carefully assess their needs, taking into account local, national, and international public health circumstances and resources; ease of access; comprehensiveness; and appropriately tailored strategies in their online communications.