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Avian malaria, ecological host traits and mosquito...
Avian malaria, ecological host traits and mosquito abundance in southeastern Amazonia
Published online by Cambridge University Press: 27 March 2017
ALAN FECCHIO ,
VINCENZO A. ELLIS ,
JEFFREY A. BELL ,
CHRISTIAN B. ANDRETTI ,
FERNANDO M. D'HORTA ,
ALLAN M. SILVA ,
VASYL V. TKACH and
JASON D. WECKSTEIN
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ALAN FECCHIO
Department of Ornithology, Academy of Natural Sciences of Drexel University, Philadelphia, PA 19103, USA
VINCENZO A. ELLIS
Departamento de Biologia Geral, Universidade Federal de Minas Gerais, Belo Horizonte, MG 31270-901, Brazil
JEFFREY A. BELL
Department of Biology, University of North Dakota, Grand Forks, ND 58201, USA
CHRISTIAN B. ANDRETTI
Curso de Pós-graduação em Zoologia, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, RS, Brazil
FERNANDO M. D'HORTA
Instituto Nacional de Pesquisas da Amazônia, Manaus, AM 69060-001, Brazil
ALLAN M. SILVA
Seção de Entomologia, Laboratório Central, Secretaria de Estado da Saúde do Paraná, São José dos Pinhais, PR 83060-500, Brazil
VASYL V. TKACH
Department of Ornithology, Academy of Natural Sciences and Department of Biodiversity, Earth, and Environmental Science, Drexel University, Philadelphia, PA 19103, USA
alanfecchio@gmail.com
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Avian malaria is a vector transmitted disease caused by Plasmodium and recent studies suggest that variation in its prevalence across avian hosts is correlated with a variety of ecological traits. Here we examine the relationship between prevalence and diversity of Plasmodium lineages in southeastern Amazonia and: (1) host ecological traits (nest location, nest type, flocking behaviour and diet); (2) density and diversity of avian hosts; (3) abundance and diversity of mosquitoes; and (4) season. We used molecular methods to detect Plasmodium in blood samples from 675 individual birds of 120 species. Based on cytochrome b sequences, we recovered 89 lineages of Plasmodium from 136 infected individuals sampled across seven localities. Plasmodium prevalence was homogeneous over time (dry season and flooding season) and space, but heterogeneous among 51 avian host species. Variation in prevalence among bird species was not explained by avian ecological traits, density of avian hosts, or mosquito abundance. However, Plasmodium lineage diversity was positively correlated with mosquito abundance. Interestingly, our results suggest that avian host traits are less important determinants of Plasmodium prevalence and diversity in southeastern Amazonia than in other regions in which they have been investigated.
Culicidae Haemosporidian parasites mosquito diversity parasite diversity Plasmodium vectors
Parasitology , Volume 144 , Issue 8 , July 2017 , pp. 1117 - 1132
DOI: https://doi.org/10.1017/S003118201700035X[Opens in a new window]
Copyright © Cambridge University Press 2017
Many factors have been proposed to explain parasite diversity (Poulin, Reference Poulin1997). For example, previous studies have shown that species richness increases towards the equator for some groups of parasitic organisms (Rohde and Heap, Reference Rohde and Heap1998; Guernier et al. Reference Guernier, Hochberg and Guégan2004; Nunn et al. Reference Nunn, Altizer, Sechrest and Cunningham2005). However, a recent meta-analysis of 62 studies involving animal, plant and fungal hosts showed that there was no strong evidence for an effect of latitude on parasite species richness (Kamiya et al. Reference Kamiya, O'Dwyer, Nakagawa and Poulin2014). Parasite diversity might be determined by characteristics of hosts rather than those of the environment. For example, host body size, population density and geographic range have all been suggested as universal predictors of variation in parasite species richness (Kamiya et al. Reference Kamiya, O'Dwyer, Nakagawa and Poulin2014). Nevertheless, the meta-analysis of Kamiya et al. (Reference Kamiya, O'Dwyer, Nakagawa and Poulin2014) did not include vector-transmitted parasites. Host density can be especially important for parasites, which depend on hematophagous insects for reproduction because the concentration of hosts in a given area potentially affects the prevalence and transmission of vector-borne pathogens by influencing encounter rates between vectors and susceptible hosts (Nunn and Heymann, Reference Nunn and Heymann2005).
Avian malaria is a worldwide, vector-transmitted disease caused by haemosporidian parasites in the genus Plasmodium (Valkiūnas, Reference Valkiūnas2005). These parasites reproduce sexually in female mosquito vectors from the genera Culex, Aedes, Culiseta, Anopheles, Mansonia, Aedeomyia and Coquillettidia (Diptera: Culicidae) (Valkiūnas, Reference Valkiūnas2005; Njabo et al. Reference Njabo, Cornel, Sehgal, Loiseau, Buermann, Harrigan, Pollinger, Valkiūnas and Smith2009; Santiago-Alarcon et al. Reference Santiago-Alarcon, Palinauskas and Schaefer2012). Environmental factors, especially temperature, can play a role in the distribution, prevalence and transmission of these parasites (Gonzalez-Quevedo et al. Reference Gonzalez-Quevedo, Davies and Richardson2014; Oakgrove et al. Reference Oakgrove, Harrigan, Loiseau, Guers, Seppi and Sehgal2014). Temperature constrains not only parasite sporogonic development (LaPointe et al. Reference LaPointe, Goff and Atkinson2010), but also influences the activity and development of the mosquito vectors, which are important determinants of the prevalence and transmission of avian malaria. However, temperature, among other environmental variables, did not explain avian malaria prevalence in avian species across several sites in South Africa (Okanga et al. Reference Okanga, Cumming and Hockey2013). As Okanga et al. (Reference Okanga, Cumming and Hockey2013) pointed out, this suggests that the prevalence of avian malaria parasites may also be determined by factors related to their avian hosts and mosquito vectors (see also Ellis et al. Reference Ellis, Collins, Medeiros, Sari, Coffey, Dickerson, Lugarini, Stratford, Henry, Merrill, Matthews, Hanson, Roberts, Joyce, Kunkel and Ricklefs2015).
Several studies have investigated the effect of avian ecological traits on the probability of infection by Plasmodium and other related haemosporidians (Ricklefs et al. Reference Ricklefs, Swanson, Fallon, Martinez-Abrain, Scheuerlein, Gray and Latta2005; Fecchio et al. Reference Fecchio, Lima, Silveira, Braga and Marini2011, Reference Fecchio, Lima, Svensson-Coelho, Marini and Ricklefs2013; Svensson-Coelho et al. Reference Svensson-Coelho, Blake, Loiselle, Penrose, Parker and Ricklefs2013; González et al. Reference González, Matta, Ellis, Miller, Ricklefs and Gutiérrez2014; Lutz et al. Reference Lutz, Hochachka, Engel, Bell, Tkach, Bates, Hackett and Weckstein2015; Matthews et al. Reference Matthews, Ellis, Hanson, Roberts, Ricklefs and Collins2016). In these studies, variation in haemosporidian prevalence is thought to be a result of the host's capability to resist and control infection or the result of differential exposure to parasites. However, most studies have not considered the role of vectors in explaining these patterns (but see Medeiros et al. Reference Medeiros, Ricklefs, Brawn and Hamer2015).
At a finer spatial scale, infection risk for Plasmodium in blue tit populations increased with increasing proximity to a large water source, possibly as result of increased vector abundance (Wood et al. Reference Wood, Cosgrove, Wilkin, Knowles, Day and Sheldon2007). Haemosporidian prevalence was also higher in the wettest of two western Amazonian 100 ha forest plots that were otherwise similar with respect to forest type, altitude, human disturbance and flooding (Svensson-Coelho et al. Reference Svensson-Coelho, Blake, Loiselle, Penrose, Parker and Ricklefs2013). These studies corroborate the idea that prevalence of blood parasites is higher at sites where vectors are more abundant.
Besides vector abundance, avian host density could also play a role in determining the prevalence of avian malaria parasites. For example, host population density can influence the spread and distribution of parasites by increasing the probability that the vectors and thus the parasites can come into contact with hosts (Anderson and May, Reference Anderson and May1978; Ellis et al. Reference Ellis, Medeiros, Collins, Sari, Coffey, Dickerson, Lugarini, Stratford, Henry, Merrill, Matthews, Hanson, Roberts, Joyce, Kunkel and Ricklefs2017). Furthermore, Drovetski et al. (Reference Drovetski, Aghayan, Mata, Lopes, Mode, Harvey and Voelker2014) found that haemosporidian lineages infected abundant bird species more frequently than less common host species in four avian communities in Africa, Asia and Europe.
To improve our understanding of avian malaria transmission, we sought to determine which biological factors (mosquito abundance and diversity; host ecological traits and density) explain the prevalence and diversity of Plasmodium lineages across seven locations along the Tapajós and Jamanxim rivers in Brazilian Amazonia. Specifically, we predicted that: (1) Plasmodium prevalence in a given host species would be positively correlated with the host species' density; (2) Plasmodium lineage diversity would be positively correlated with avian host diversity; (3) avian ecological traits would explain some of the variation in Plasmodium prevalence; and (4) the prevalence and diversity of avian Plasmodium lineages would be positively correlated with the abundance and diversity of mosquitoes. A secondary motivation of our study was to provide information on the diversity and distribution of ornithophilic mosquitoes in a region of Amazonia that has not previously been explored with respect to these insects.
Sampling sites
The study area was located midway down the Tapajós River, a major south bank tributary of the Amazon River. Sampling was carried out along both banks of the Tapajós River covering both the Rondônia and Tapajós areas of endemism (Silva et al. Reference Silva, Rylands and Fonseca2005). We also sampled the right bank of its most important tributary, the Jamanxim River (Fig. 1). The area comprises a wide variety of microhabitats, with the terra firme and igapó forests being the most broadly distributed. Sampling was conducted along six transects of five km in length, beginning in the seasonally flooded forest (igapó) and crossing the interior of terra firme forest. We also sampled a 250 m transect on a river island. We named these seven sites as follows: the first letter of the label is the first letter of the river name ('T' for Tapajós and 'J' for Jamanxim); the second letter is the margin of the river ('L' for left bank, 'R' for right bank and 'I' for the island); site names also include a number to identify unique sampling sites on the same bank of the same river.
Fig. 1. Map showing the seven sampling sites along the Tapajós and Jamanxim rivers. Background shading corresponds to elevation with lower areas represented by darker shading. The inset shows the state of Pará, Brazil, and the study area (SA) is represented as a dark grey rectangle within the state. Am, Amazon River; Jam, Jamanxim River; and Tap, Tapajós River.
Bird sampling
We placed five mist net lines within each transect. These five mist net lines were each separated by 1 km and contained ten nets. The 50 mist nets (12 m length × 3 m height) remained open for three consecutive mornings at each of the seven transect sites. Owing to the small size of the river island, the mist nets were arranged in a single 250 m line and were sampled with the same effort as the others sites. Blood sampling of captured birds took place during two distinct seasons, the dry season for the sites TL1, TL2, TL4, TR1 and JR1 (18 July–3 August 2012) and flooding period for the sites TL1, TL2, TL3 and TI (1–17 October 2012), thus only two sites, TL1 and TL2, were sampled during both the dry and flooding seasons. At each site, to allow sufficient time for transmission of Plasmodium to avian hosts, avian blood samples were collected from individual birds shortly after (~2–3 weeks) mosquito vectors were also sampled. Netted birds were bled by brachial venipuncture using heparinized capillary tubes. Blood samples were stored in 95% ethanol until DNA extraction. After blood collection, birds were ringed and released, or euthanized and prepared for museum specimens. All tissue samples and birds were collected or ringed under appropriate permits from Brazil (IBAMA no 22/2012 and ICMBio no 004/2012). Tissue samples and voucher specimens were deposited in the Bird Collection at the Instituto Nacional de Pesquisas da Amazônia – INPA, Manaus, Brazil. Species nomenclature follows Piacentini et al. (Reference Piacentini, Aleixo, Agne, Maurício, Pacheco, Bravo, Brito, Naka, Olmos, Posso, Silveira, Betini, Carrano, Franz, Lees, Lima, Pioli, Schunck, Amaral, Bencke, Cohn-Haft, Figueiredo, Straube and Cesari2015).
Besides mist net sampling, we conducted point count surveys (10 min each), spread along transects at every 500 m, totalling 11 points per transect. Each point was sampled for four consecutive days at each sampling period. We estimated density of individual species in each area using MCDS (Multiple Covariates Distance Sampling) as implemented in the program Distance 6.0 (Thomas et al. Reference Thomas, Laake, Rexstad, Strindberg, Marques, Buckland, Borchers, Anderson, Burnham, Burt, Hedley, Pollard, Bishop and Marques2009). The analyses were stratified to obtain density estimates for each area. We truncated 10% of data with larger distances within each species to avoid double counting the same individual, as recommended by Buckland et al. (Reference Buckland, Anderson, Burnham, Laake, Borchers and Thomas2001). For each species, we compared the following models: (1) half normal and hazard rate set as functions of expansion adjusted by cosine; (2) simple polynomial; and (3) polynomial hermite. We then calculated the Akaike's Information Criterion (AIC) for each model and chose the one with the lowest AIC score.
Mosquito sampling
Mosquitoes were collected using light traps powered by 12 V batteries as designed by Falcão (Reference Falcão1981). The traps were installed in the same sampling transects used for mist netting birds, and were distributed in two vertical layers, at ground level (suspended approximately 1 m above the ground) and in the forest canopy (suspended between 10 and 15 m above the ground). A total of 14 traps were installed simultaneously in each transect and these were located at equidistant points starting at the river bank and ending at 4 km from the river bank in the interior of terra firme forest. Mosquito collections were made during three consecutive nights, between 18:00 and 21:00 h during both the dry season for the sites TL1, TL2, TL4, TR1 and JR1 (2–13 July 2012) and the flooding season for the sites TL1, TL2, TL3 and TI (12–23 September 2012), thus only two sites, TL1 and TL2, were sampled during both the dry and flooding seasons. Mosquitoes were stored in small plastic vials labelled with the corresponding data for each sample. In the laboratory, the female mosquitoes were separated and identified taxonomically using external morphological characters observed using a stereoscope. We only used female mosquitoes for abundance estimation because only females are potential vectors for Plasmodium. For species determination, we used keys published by Galindo et al. (Reference Galindo, Blanton and Peyton1954) and Forattini (Reference Forattini1962, Reference Forattini1965a , Reference Forattini b , Reference Forattini2002). We used the infrageneric classification scheme of the genus Anopheles from McKeon et al. (Reference McKeon, Moreno, Sallum, Povoa and Conn2013), Moreno et al. (Reference Moreno, Bickersmith, Harlow, Hildebrandt, McKeon, Silva-do-Nascimento, Loaiza, Ruiz, Lourenço-de-Oliveira, Sallum, Bergo, Fritz, Wilkerson, Linton, Juri, Rangel, Póvoa, Gutiérrez-Builes, Correa and Conn2013) and Ruiz-Lopez et al. (Reference Ruiz-Lopez, Wilkerson, Ponsonby, Herrera, Sallum, Velez, Quiñones, Flores-Mendoza, Chadee, Alarcon, Alarcon-Ormasa and Linton2013), which differentiates among groupings of cryptic but presently unnamed lineages. The abbreviation of genera and subgenera follows the guidelines suggested by Reinert (Reference Reinert2001), according to the newly proposed mosquito nomenclature of the Walter Reed Biosystematics Unit, Smithsonian Institution (catalogue available at http://www.mosquitocatalog.org/taxon_table.aspx) and Harbach (Reference Harbach2013) in the Mosquito Taxonomic Inventory (www.mosquito-taxonomic-inventory.info/).
Parasite detection
DNA was extracted from avian blood samples using the Qiagen DNeasy 96 Blood and Tissue kit (Qiagen, Valencia, CA), following the Qiagen protocol for blood in 95% ethanol. Total DNA was screened by real-time PCR to detect haemosporidian DNA following the protocol of Bell et al. (Reference Bell, Weckstein, Fecchio and Tkach2015). Positive and negative controls were included in all real-time PCR runs. Samples identified as positive by real-time PCR underwent subsequent nested PCR, as outlined in Bell et al. (Reference Bell, Weckstein, Fecchio and Tkach2015), to amplify a 477 bp fragment of the cytochrome b gene.
Positive nested PCR products were purified using ExoSAP-IT (Affymetrix, Santa Clara, CA), sequenced using BigDye terminator v3.1 cycle sequencing kit (Applied Bio systems, Foster City, CA), and run on an ABI 3100 DNA sequencer (Applied Bio systems, Foster City, CA). For sequencing protocol and primers see Bell et al. (Reference Bell, Weckstein, Fecchio and Tkach2015). Forward and reverse sequences were visualized and assembled using Sequencher v.5.0.1 (Gene Codes Corp., Ann Arbor, MI). Chromatograms that showed the presence of multiple infections were scored as co-infections. Co-infections were separated using the program PHASE 2.1.1 (Stephens et al. Reference Stephens, Smith and Donnelly2001; Stephens and Donnelly, Reference Stephens and Donnelly2003) following the protocol of Harrigan et al. (Reference Harrigan, Sedano, Chasar, Nguyen, Whitaker and Smith2014). Assembled sequences were aligned using BioEdit v7.2.0 (Hall, Reference Hall1999) and collapsed to unique haplotypes using the FaBox haplotype collapser and converter tool (Villesen, Reference Villesen2007). Sequence identities were verified with a local BLAST against the MalAvi database (Bensch et al. Reference Bensch, Hellgren and Pérez-Tris2009) using BioEdit v7.2.0 (Hall, Reference Hall1999). New lineages were named after the host of origin following standard protocol (Bensch et al. Reference Bensch, Hellgren and Pérez-Tris2009), using a six-letter code produced by using the first three letters of both the host genus and specific epithet followed by a number to denote multiple lineages from a single host species. For example, lineage WILPOE01 represents the first lineage obtained from Willisornis poecilinotus. All sequences were deposited in GenBank (Accession No KU562250–KU562512) and the MalAvi database.
Assembled sequences of unique lineages were used to reconstruct a molecular phylogeny using Bayesian inference (BI) as implemented in MrBayes v. 3.2.2 (Huelsenbeck and Ronquist, Reference Huelsenbeck and Ronquist2001; Ronquist and Huelsenbeck, Reference Ronquist and Huelsenbeck2003) and the GTR + I + G model of nucleotide substitution as determined by jModelTest (Darriba et al. Reference Darriba, Taboada, Doallo and Posada2012); Leucocytozoon fringillarum (FJ168564) served as the outgroup. The BI analysis was run until the s.d. of split frequencies stabilized below 0·01. Twenty-five percent of resulting trees were discarded as burn in. The resulting consensus tree was visualized in FigTree (Rambaut, Reference Rambaut2009).
Statistical analyses and modelling
We were interested in testing whether the prevalence of Plasmodium parasites was related to the density of avian hosts within the different sites sampled. To do this we first restricted our analysis to species sampled at least five times within a site; this left us with three sites (TL1, TL2 and TL3) each including more than three species that met our sample size criterion (Table A2). We then calculated prevalence for each species at each site separately as the number of Plasmodium-infected individuals divided by the total number of individuals sampled of a particular species at a particular site. Individuals infected with only Haemoproteus parasites were considered uninfected for this analysis since we did not sample the arthropod vectors for this parasite genus and because birds can be found with mixed infections of both genera (Valkiūnas, Reference Valkiūnas2005). Individual birds infected with both Plasmodium and Haemoproteus were considered infected. We then ran three generalized linear models, one for each site, with prevalence as the response variable and host density as the explanatory variable with a quasibinomial error structure to account for overdispersion (Bolker et al. Reference Bolker, Brooks, Clark, Geange, Poulsen, Stevens and White2009). We entered two vectors of infected and uninfected individuals into the model to weight prevalence by sample size as is standard when running such models in R; see Crawley, Reference Crawley2012). Since sampling took place in two seasons for sites TL1 and TL2, we compared prevalence of Plasmodium parasites between seasons (within sites; prevalence was calculated for all individuals, i.e. not separated by host species, sampled within each season for each of the two sites) using chi-squared tests to confirm that seasons could be grouped for this analysis.
We tested the hypothesis that Plasmodium lineage diversity was related to the diversity of avian hosts across all seven sampling sites. For this we calculated a Simpson's index of diversity for all avian hosts that were sampled (117 species) and one for all parasite lineages within each sampling site using the 'diversity' function in the R package vegan (Oksanen et al. Reference Oksanen, Blanchet, Kindt, Legendre, Minchin, O'Hara, Simpson, Solymos, Stevens and Wagner2015). We then used Spearman's rank correlation tests (correlation statistic is ρ) to determine whether the variables were correlated across sampling sites.
We were also interested in testing whether the prevalence of Plasmodium parasites across avian host species was related to the following host ecological variables: nest location (ground, understory, sub canopy, canopy and cliff/bank), nest type (open cup, closed cup and cavity), flocking behaviour (solitary/family, single species and mixed species), and diet (insectivore, frugivore/granivore and omnivore). We scored these traits for all species sampled using a combination of The Birds of South America Volumes I and II (Ridgely and Tudor, Reference Ridgely and Tudor1989a , Reference Ridgely and Tudor b ), Neotropical Birds: Ecology and Conservation (Stotz et al. Reference Stotz, Fitzpatrick, Parker and Moskovits1996), The Cornell Lab of Ornithology: Neotropical Birds (http://www.neotropical.birds.cornell.edu/portal/home) and WikiAves (http://www.wikiaves.com.br). We pooled our site data for this analysis because: (1) we did not expect site to influence the relationship of prevalence and ecological host traits (e.g. Matthews et al. Reference Matthews, Ellis, Hanson, Roberts, Ricklefs and Collins2016); and (2) because we did not find any significant differences in the prevalences of individual species sampled at multiple sites in our study (results not shown). We therefore constructed a generalized linear model with Plasmodium prevalence (weighted by sample size) as the response variable and each of the ecological variables as explanatory variables. We ran the model with a quasibinomial error structure to account for overdispersion and only included species with at least five individuals sampled (n = 44 avian species). Initially we included the taxonomic family of host species as an explanatory variable in the model to account for potential differences in prevalence among families, but sparse sampling led to uninterpretable estimates of family-level prevalences and so we dropped family from our final model. We also ran Wald chi-squared tests on each of the explanatory variables in the model using the function 'wald.test' in the R package aod (Lesnoff and Lancelot, Reference Lesnoff and Lancelot2012).
Finally, we tested the hypothesis that the prevalence and diversity of Plasmodium lineages were positively related to the abundance and diversity of mosquitoes across all seven of our sampling sites using Spearman's rank correlation tests. Here we calculated parasite prevalence for sites rather than for species by dividing the total number of infected individuals by the total number of individuals sampled in a site irrespective of host species. We again calculated a Simpson's index of diversity for Plasmodium lineages and one for mosquitoes within each sampling site.
All statistical analyses were performed in R version 3.2.3 (R Core Team, 2015).
Prevalence and diversity of Plasmodium lineages
We analysed 675 birds of 120 species sampled in seven communities along the Tapajós and Jamanxim rivers (Fig. 1, Table A1). Plasmodium infections were detected in 136 individuals from 51 host species with a prevalence of 20% (Table A1). Infection prevalence varied among well sampled host species (>10 individuals screened), ranging from 0 to 64% (Table A1). Plasmodium prevalence was homogeneous across sites, ranging from 12 to 31% (χ 2 = 11·282, d.f. = 6, P = 0·080; Table 1). Based on cytochrome b divergence, we recovered 89 haemosporidian lineages within the genus Plasmodium, of which 81 (91%) were reported for the first time (Fig. 2). Although there are several well resolved and supported clades within the phylogeny, the general pattern is one of many polytomies with low node support (Fig. 2). When mapped onto the phylogeny, host family shows little perceivable pattern within the phylogeny, dominated by the highly sampled family Thamnophilidae. In several cases, individual Plasmodium lineages were recovered from more than one host family, with three Plasmodium lineages found in individual hosts from four different host families.
Fig. 2. BI phylogenetic reconstruction of Plasmodium lineages recovered from sites along the Tapajόs and Jamanxim rivers. Posterior probability support above 0·9 is noted at the base of nodes and host family is noted next to terminal taxon labels.
Table 1. Prevalence of Plasmodium per site along the Tapajós and Jamanxin rivers, southeastern Amazonia, Brazil. Specific site location information can be found in Fig. 1
We sampled birds at sites TL1 and TL2 during the dry and flooding periods, allowing us to test for seasonal differences in prevalence. We found no differences in overall prevalence of Plasmodium parasites between the two seasons within each site (site TL1, χ 2 = 0, d.f. = 1, P = 1; site TL2, χ 2 = 0·703, d.f. = 1, P = 0·402). For these tests we used ten host species from site TL1 and nine host species from the site TL2 for which we had sampled at least five individuals. We also found no differences in prevalence between seasons for individual host species (results not reported).
Avian host ecology and Plasmodium prevalence
Prevalence of Plasmodium (calculated for each host species within sites) was positively related to host density at site TL3 (GLM coefficient = 0·186 ± 0·05 s.e., P = 0·037, n = 5), but not at sites TL1 (0·027 ± 0·09 s.e., P = 0·769, n = 8) and TL2 (−0·096 ± 0·20 s.e., P = 0·661, n = 6). However, the relationship at site TL3 was based on only five host species, and would not be considered significant after a Bonferroni correction (after three tests, alpha = 0·05/3 or 0·017; Table A2). The diversity of Plasmodium parasites was not related to the diversity of avian hosts (ρ = 0·49, P = 0·268, n = 7).
We found no relationship between ecological traits of the hosts and the prevalence of Plasmodium parasites. We report coefficients and significance of each category of the explanatory ecological variables in Table 2 (results of each host trait modelled separately can be found in Table A3). We also ran Wald chi-squared tests for each ecological variable across all coefficients and none were significant (nest location: χ 2 = 1·4, d.f. = 4, P = 0·85; nest type: χ 2 = 4·6, d.f. = 2, P = 0·10; flocking behaviour: χ 2 = 1·6, d.f. = 2, P = 0·45; diet: χ 2 = 2·1, d.f. = 2, P = 0·35). For this analysis, we used 504 birds of 44 species with a minimum of five individuals sampled per species. Plasmodium infections were detected in 101 individuals from 23 host species with a prevalence of 20% (Table A4).
Table 2. The results of a generalized linear model relating four avian ecological traits to the prevalence of Plasmodium parasites in avian hosts
All of the explanatory ecological variables were categorical. We therefore report the estimate of the coefficient of each of the levels of those variables in relation to a base level. The base levels are as follows: nest location is ground, nest type is open cup, flocking is solitary/family, diet is insectivore. We report the estimate of the coefficient for each variable, its s.e., t value and P value; the null deviance of the model is 198·01 on 43 d.f. and the residual deviance is 128·35 on 33 d.f.
Mosquitos, prevalence and diversity of Plasmodium lineages
We collected 511 female mosquitoes from 56 species and morpho-species belonging to 11 genera (Tables A5 and A6). We ran paired Mann–Whitney U tests to compare mosquito abundance between seasons in sites TL1 and TL2. In site TL1, mosquito abundance was higher in the flooding period than in the dry season (U = 21·5, P < 0·001), but in site TL2, there was no difference in mosquito abundance between seasons (U = 169, P = 0·441). In the other five sites, mosquito collection took place in only one season, preventing further seasonal comparisons (Tables A5 and A6).
The prevalence of Plasmodium parasites in avian hosts (calculated for an entire site and not by host species) was not related to the diversity (ρ = −0·46, P = 0·302) or the abundance (ρ = −0·22, P = 0·641) of mosquitoes across the seven sampling sites. The diversity of Plasmodium parasites was also not related to the diversity of mosquitoes (ρ = 0·67, P = 0·102), but was positively related to the abundance of mosquitoes (ρ = 0·79, P = 0·034) across sampling sites (Fig. 3). Since each of these tests represents a separate hypothesis, we did not apply a Bonferroni correction to the resulting P values.
Fig. 3. Relationship between the diversity of Plasmodium parasites (calculated using Simpson's index of diversity) and mosquito abundance at each of the sampled sites; point size is scaled to the number of individual birds sampled at each site. The two variables are positively correlated (ρ = 0·79, P = 0·034).
We investigated biotic factors that may determine the prevalence and diversity of Plasmodium parasites in Amazonian birds. We found that the diversity of Plasmodium lineages was positively correlated with mosquito abundance across the seven bird communities we sampled. Neither bird density nor bird diversity explained prevalence or lineage diversity of Plasmodium parasites among avian hosts. None of the four ecological traits of avian hosts explained Plasmodium prevalence. The lack of seasonal differences in prevalence of Plasmodium found along the Tapajós River needs to be considered with caution since we tested this with samples collected from only two communities.
Our finding that Plasmodium lineage diversity is correlated with mosquito abundance and not with any avian host traits suggests that diversity and distribution of these parasites might be constrained by the final host (Culicidae) in our bird–parasite–mosquito system. Ishtiaq et al. (Reference Ishtiaq, Guillaumot, Clegg, Phillimore, Black, Owens, Mundy and Sheldon2008) demonstrated that the movement of Plasmodium lineages among southwest Pacific Islands might be restricted by the lack of overlap in the distributions of competent vector species. One possibility is that the mosquito community in our study region only has a few competent mosquito vectors and their signal was washed out by the huge mosquito diversity found in southeastern Amazonia. Alternatively, regional processes at the level of host populations, such as immunity to particular parasite lineages or differential exposure to certain parasite lineages, might mask any density-dependent influence of avian hosts and mosquitoes vectors on prevalence. For instance, the distribution and diversity of many parasites among host populations are known to be highly variable and one of the main reasons behind this is the inequality of individual host immune responses in defending themselves against particular parasites (Poulin, Reference Poulin2007).
Our results demonstrate that Plasmodium lineage diversity and prevalence in Amazonian birds does not vary with host ecological traits and avian host density. We expected that host population density would be positively correlated to avian malaria prevalence since it affects vector–host-encounter rates (Dobson, Reference Dobson2004). Although several studies have found evidence for higher haemosporidian prevalence in denser host populations (Matthews et al. Reference Matthews, Ellis, Hanson, Roberts, Ricklefs and Collins2016; Ricklefs et al. Reference Ricklefs, Soares, Ellis and Latta2016; Ellis et al. Reference Ellis, Medeiros, Collins, Sari, Coffey, Dickerson, Lugarini, Stratford, Henry, Merrill, Matthews, Hanson, Roberts, Joyce, Kunkel and Ricklefs2017), many others failed to find such an association (Svensson-Coelho et al. Reference Svensson-Coelho, Blake, Loiselle, Penrose, Parker and Ricklefs2013; Gonzalez-Quevedo et al. Reference Gonzalez-Quevedo, Davies and Richardson2014). Kilpatrick et al. (Reference Kilpatrick, Daszak, Jones, Marra and Kramer2006) showed that West Nile virus transmission within a local host community was influenced by extreme heterogeneity in mosquito feeding patterns. At least in some sites, transmission of multi-host pathogens such as avian malaria, may be influenced by heterogeneity in host–vector compatibility more than by bird density (Kilpatrick et al. Reference Kilpatrick, Daszak, Jones, Marra and Kramer2006; Medeiros et al. Reference Medeiros, Hamer and Ricklefs2013). Specifically, for avian malaria, higher abundance of vectors does not lead to a higher host–vector-encounter rate. For example, Medeiros et al. (Reference Medeiros, Ricklefs, Brawn and Hamer2015) showed that vectors overutilized some bird species regardless of their abundance and Plasmodium prevalence may be associated with vector utilization rather than vector abundance.
Nest characteristics among many other ecological traits of host individuals might be associated with variation in haemosporidian prevalence (Ricklefs et al. Reference Ricklefs, Swanson, Fallon, Martinez-Abrain, Scheuerlein, Gray and Latta2005; Fecchio et al. Reference Fecchio, Lima, Silveira, Braga and Marini2011, Reference Fecchio, Lima, Svensson-Coelho, Marini and Ricklefs2013; Svensson-Coelho et al. Reference Svensson-Coelho, Blake, Loiselle, Penrose, Parker and Ricklefs2013; González et al. Reference González, Matta, Ellis, Miller, Ricklefs and Gutiérrez2014; Lutz et al. Reference Lutz, Hochachka, Engel, Bell, Tkach, Bates, Hackett and Weckstein2015; Matthews et al. Reference Matthews, Ellis, Hanson, Roberts, Ricklefs and Collins2016). However, based on simple correlations, these studies relied on the premise that nestlings or adults are more exposed to the vectors in the nest according to its architecture. Moreover, mixed results found in these studies suggest that the relationship between nest type and risk of infection by haemosporidian parasites might be location dependent. An analysis of the identity of host blood found in engorged female mosquitoes could provide a general test for this pattern and confirm whether host nest type can predict haemosporidian prevalence in birds.
Lack of seasonal variation in Plasmodium prevalence found in two communities sampled along the Tapajós River confirms the temporal stability of these parasites in tropical birds. There are three main possible explanations for this: (1) the abundance of mosquito vectors might differ between seasons (i.e. site TL1), but even the lower number of actively feeding mosquitoes is sufficient to ensure transmission especially if these few mosquitoes are the most competent vector for more prevalent Plasmodium lineages; (2) infections in birds last long enough to span more than a single season and thus mask the changes in mosquito abundance between seasons; and (3) vector abundance might be stable over seasons (i.e. site TL2), allowing Plasmodium transmission throughout the year. If Plasmodium parasites have a dynamic aspect in tropical bird communities, it may vary with years or decades and not between seasons within the same year.
Several species of mosquito belonging to the genera Aedeomyia, Anopheles, Coquillettidia, Culex, Culiseta, Mansonia, Aedes (Ochlerotatus and Stegomyia) have been implicated in the transmission of Plasmodium spp. in birds (Valkiūnas, Reference Valkiūnas2005; Njabo et al. Reference Njabo, Cornel, Sehgal, Loiseau, Buermann, Harrigan, Pollinger, Valkiūnas and Smith2009). Among the species of mosquito inhabiting our study area in Tapajos, Ad. (Ady.) squamipennis is known as a natural vector of avian malaria in Venezuela (Gabaldon et al. Reference Gabaldon, Ulloa and Pulido1981). Another Neotropical mosquito Culex (Melanoconion) ocossa, together with Ad. (Ady.) squamipennis could be responsible for the transmission of avian malaria in some regions of Panama (Loaiza and Miller, Reference Loaiza and Miller2013). Unfortunately, we were not able to analyse the engorged females of these mosquitoes to study vectorial capacity or host specificity and thus future research on this is necessary as a first step to fully understand the transmission risk and high diversity of avian malaria parasites in the Neotropical region.
The absence of any perceivable pattern of host family within the phylogeny can be attributed to both the high level of sampling from the host family Thamnophilidae and the low level of host specificity known for Plasmodium (Beadell et al. Reference Beadell, Gering, Austin, Dumbacher, Peirce, Pratt, Atkinson and Fleischer2004, Reference Beadell, Covas, Gebhard, Ishtiaq, Melo, Schmidt, Perkins, Graves and Fleischer2009; Valkiūnas, Reference Valkiūnas2005; Dimitrov et al. Reference Dimitrov, Zehtindjiev and Bensch2010; Ishtiaq et al. Reference Ishtiaq, Clegg, Phillimore, Black, Owens and Sheldon2010). This low level of host specificity is shown in those lineages recovered from three and four different host families, spanning different host orders. Host switching is an important evolutionary mechanism in avian haemosporidian parasites with closely related haemosporidian lineages conserved within higher host taxa (Waldenström et al. Reference Waldenström, Bensch, Kiboi, Hasselquist and Ottoson2002; Križanauskiené et al. Reference Križanauskiené, Hellgren, Kosarev, Sokolov, Bensch and Valkiūnas2006; Ricklefs et al. Reference Ricklefs, Outlaw, Svensson-Coelho, Medeiros, Ellis and Latta2014). Due to high levels of host switching and subsequent dispersal, cospeciation is not thought to have played a large role in the evolutionary history of Plasmodium (Ricklefs et al. Reference Ricklefs, Outlaw, Svensson-Coelho, Medeiros, Ellis and Latta2014; Lauron et al. Reference Lauron, Loiseau, Bowie, Spicer, Smith, Melo and Sehgal2015). However, the high host diversity in Amazonia would be an ideal system to explore the existence of coevolutionary links between Plasmodium parasites and avian hosts in future studies.
The heterogeneous prevalence of Plasmodium across bird species in southeastern Amazonia, regardless of their ecological traits, suggests that constraints on the distribution of these parasites are related to vectors within these assemblages. This highlights the importance of exposure to vectors in explaining avian malaria prevalence (Medeiros et al. Reference Medeiros, Ricklefs, Brawn and Hamer2015). Nonetheless, the results have to be interpreted cautiously, because of low sample sizes within sites and the low number of sites in general. Therefore, the next step in understanding the factors promoting the high diversity and heterogeneity of Plasmodium lineages in this region of Amazonia, as well as the mechanisms that produce variation in the prevalence of these vector-borne parasites across avian hosts, must include studies that integrate rates of vector exposure, feeding preference and vectorial capacity of the mosquitoes in the same area.
We thank the ornithologists: Marcelo Barreiros, Gabriel Leite, Thiago Bicudo, Thiago Moura, Daniel Gressler, Camila Duarte and several Munduruku Indians who helped collect the blood samples. We also thank Camila Ribas and Mario Cohn-Haft from the Bird and Genetic Resources Collections of Instituto Nacional de Pesquisas da Amazônia who loaned the samples used in this study. We also thank three anonymous referees for their comments on earlier versions of the manuscript and Vitor Piacentini who kindly reviewed the bird nomenclature.
This work was funded by the Fundação de Amparo à Pesquisa do Estado do Amazonas – FAPEAM and Conselho Nacional de Desenvolvimento Científico e Tecnológico – CNPq (Process number 61.0012/2008.7-DCR/CNPq/FAPEAM) to A.F. and US National Science Foundation grants DEB-1503804 to J.D.W. and DEB-1120734 to V.V.T. During the project, A.F. was supported by a postdoctoral fellowship from CNPq (Process numbers 350140/20120 and 201275/2014-7) and he is currently maintained by a PNPD scholarship from Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES).
Table A1. Plasmodium prevalence from seven bird communities in southeastern Amazonia, Brazil. Species of birds are organized taxonomically
Table A2. The 11 bird species used to test the effect of host density (individuals per 100 ha) on prevalence of Plasmodium from three transects along the Tapajós River. The geographic locations of sites can be found in Fig. 1
Table A3. In the main text of our paper we report the results of a multiple regression (GLM) modelling prevalence as a function of four explanatory variables (nest location, nest type, flocking and diet). Here we report the results of four separate models of each explanatory variable by itself, for comparison. Each model is a GLM with a 'quasibinomial' error structure (to account for overdispersion as in the model presented in the main text); coefficients are reported as in Table 2 of the main text
Table A4. The 44 species used in the analysis of an effect of avian ecological traits on Plasmodium prevalence in southeastern Amazonia
Table A5. Mosquito species caught across five sites during the dry season. The geographic locations of sites can be found in Fig. 1
Table A6. Mosquito species caught across four sites during the flooding season. The geographic locations of sites can be found in Fig. 1
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ALAN FECCHIO (a1), VINCENZO A. ELLIS (a2), JEFFREY A. BELL (a3), CHRISTIAN B. ANDRETTI (a4), FERNANDO M. D'HORTA (a5), ALLAN M. SILVA (a6), VASYL V. TKACH (a3) and JASON D. WECKSTEIN (a7)
DOI: https://doi.org/10.1017/S003118201700035X
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Action resumes onmatch day 6 of the 2019 AFCON qualifiers with teams vying for places in the competition proper.
Nigeria Super Eagles will not be left out as they face Pirates of Seychelles in a dead rubber match.
Despite the nature of the game, Nigeria's Coach, Gernot Rohr doesn't want to leave any unturn as he invited first team players for the match amidst noise that the fringe and home based players should have been made use of.
Seychelles on their own are coming to Nigeria with their first team players simply because large chunks of them ply their trade at home.
The Three time African champions started their campaign on a losing note as they were beaten 0-2 by Bafana Bafana of South Africa at the Godswill Akpabio stadium in Uyo.
They rallied back on match day 2 as they walloped Seychelles 3-0 and continued to grow in strength since then.
Eyes will be on newly invited players such as FC Midtjyland striker Paul Onuachu whom the coach has confirmed that he will be giving an opportunity in the game.
Some home lads like Ndifreke Effiong and Flying Eagles duo of Valentine Ozornwaifor and skipper Ikouwem Udoh might also get playing time.
Nigeria sit a point above South Africa on top Group E, heading into this weekend's round of fixtures, and will be expected to retain the position with victory over Seychelles.


This was the first terrorist explosion in Phnom penh for nearly two years It could signal the beginning of terrorist tactics.
Background: This was the first terrorist explosion in Phnom penh for nearly two years It could signal the beginning of terrorist tactics. Bomb, believed to be plastic explosive, went off shortly after 7 am this morning, killing one early shopper in the central market place.
Police cleared the main building, fearing another possible explosion. Market went on as usual outside. Doctors arrived after 8 am and examined the body. Other officials checked the scene for bomb clues.


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She held a hand on a hilt of one of her throwing knives.
The house in front of us looked like a place where demons would reside. The whole place was black. The shutters looked like cat toys shredded and bent. Part of the roof had caved in. Bricks were cracked and jagged. Ivy covered the chimney. The top of the chimney had been blasted apart. And it was night.
I grabbed my sword and kicked open the door.
A demon stood, leering at me. Imagine a goat with red eyes and a forked tongue. Oh, and they spit acid too.
I swung my silver steel alloy sword and cut through the demon. The leering thing spit a stream of the acid they're famous for. The door frame melted, and Sabrina jumped in.
Right as the door frame collapsed, Sab made it. Our exit didn't.
Sabrina said a word not suitable for me to repeat here and then proceeded to state the obvious.
I whistled and then lit the lantern.
I heard a thunk, and a dead demon fell in front.
"Little beasts are worse than rats," said Sabrina.
I cracked the top of the bottle, pulled up the stick inside, and threw it. The stick's friction caused a spark. And this stuff is highly flammable.
The bottle exploded, sending bits of flame everywhere. The old table inside blew apart, sending old wood into the gloom. Spider webs caught on fire, and an old armchair burst into flame. The room had been a sitting room before. The fireplace mantel collapsed, bringing down a box and a small painting. A larger painting was ablaze and a fine couch on which the likes of noblewoman would have sat on would now have set their dresses on fire.
A plate of old, stale pastries was aflame. The demons were screeching as they each kicked the bucket.
I ran into the room and ran for the kitchen, located in back of the house. Several demons jumped at me, screaming with hatred. I kicked the flaming door down, ran in, grabbed an old bottle of wine, spun around, and slammed it into a demon's face.
My sword bit into demon after demon.
I started swinging, but my arm was tired. The demons understood this; trying to pull a fast one n us.
I dropped my defense and a demon jumped.
I held up my hand in a futile attempt to ward it off.
Then, my hand started glowing blue. The demon got a face full of blue orb. Several more lunged, and they each got a face full of blue light. Sabrina's wound had closed up, and the demons were gone.
"Agreed, but for good measure …" I pulled the second flame grenade off my belt.


text:
OklahombreS Online! oklahombres.org oklahombres.org General Oklahombres "Old West" Oklahombres (pre 1907) Who killed Belle Starr?
Who killed Belle Starr?
Wed September 08 2004 03:33 PM
From Jack Moseley's column in the Fort Smith Times Record (September 8, 2004):
It's true that Belle ran with the bad boys of Indian Territory, packed an ivory-handled pistol and bought her clothes at (the) Fort Smith Boston store . . . .
(Jerry) Akins, who has read hundreds of newspaper accounts, depositions from Judge Parker's court and other documents, including one that reveals that the man who some wrongly claim murdered Belle was issued the warrant to arrest the man who was actually accused of the crime.. . .
Some have argued that Blue Duck, rumored to be Belle's lover, killed her. "Impossible," says Akins.
"Blue Duck was in prison for murder at the time. He did not get out until 1895 . . . ".
Others contend that Belle was killed by her son, Eddie Reed, supposedly because she had recently whipped him for mistreating one of her horses. Highly unlikely, says Akins. "The son lived in the Cherokee Territory. Belle lived in Choctaw Territory miles away. Besides, they had not seen each other in months."
A lot of folks claim Belle was murdered by her husband whom sometimes went by Jim July and sometimes by Jim Starr. Belle had ridden halfway to Fort Smith with Jim July, camped overnight and started back on the day she was murdered. . . .
Days later, Jim was given the arrest warrent to serve on Edgar A. Watson, the man Akins believed most likely killed Belle. Watson was arrested by Jim July right after Belle was laid to rest wearing a black dress and holding her favorite pistol that was interred with her body. . . .
Watson was acquitted of killing Belle. There were no witnesses to the actual murder. He steadfastly contended he was an innocent man.
--meursault
This message has been edited. Last edited by: meursault,
Thu September 09 2004 08:05 AM
Posts: 215 | Registered: Thu December 11 2003 IP
<M. Koch>
The murder mystery of who killed Belle Starr has been going on for years. I have asked many people who lived in the area where she was killed in nearby Hoyt, Oklahoma, who they felt murdered Belle. Most have stated it was her son. The actual location of the shooting is not too hard to locate, but has probably changed through the years, as it is near the river bottoms.
Crosstimbers Okie
Sun August 05 2007 06:06 PM
Do you happen to know where the location of her murder is exactly? Is it on private land? What about Younger's Bend and her grave?
www.PrisonOfficer.Org
Posts: 26 | Location: Kansas City, MO USA | Registered: Mon October 02 2006 IP
Ignored post by Crosstimbers Okie posted
<MKoch>
Mon August 06 2007 06:18 AM
She was shot and killed by the town of Hoyt, Oklahoma, near the Canadian River in present day Haskell County. Obviously, the river has change its location since she was killed, so oneone knows exactly where she was killed, but old timers from years ago have identified a present day dirt road, which has a fork to it near the river, just north of the town of Hoyt, where she was killed. It is not on private property that I am aware of. Her spot of burial is very hard to get to during the summer since the grass is not mowed and with all the trees, you are not able to see the dam as Dee has stated previously. I do not know who presently owns the property. Good luck.
I think I've located the road on which she was shot. I don't think that the river there has moved enough to interefere with the spot of ambush as it was a respectable distance from the river and the county boundaries on that part of the river coincide pretty closely to the location of the river. Those boundaries are the same as those that separated the Cherokee & Choctaw Nations.
Perhaps this Fall I can get down and try and locate the grave site.
R Bell
I have been to the grave site many times. Let me know if I can help you on this. It is harder to find than it used to be due to an ice storm that damaged the natural trails a few years ago. I would not suggest going to it until the winter or early spring. There are many other related sites in the area also that might intereest you.
Posts: 26 | Registered: Sat February 04 2006 IP
Ignored post by R Bell posted
Great! I may be fishing Eufaula this fall. If so I would like to locate it. Can you give me directions from the dam?
What are the other sites?
I've noticed the creek that comes off of the mountain and empties into the Canadian about a mile east of the dam. I've always wondered if that creek would take one up to Younger's Bend. My biggest concern is that I don't want to upset a landowner.
PFF
Tue October 28 2008 04:36 AM
Blueford "Blue" Duck's story is at http://www.westernoutlaw.com/gravesites/bluford_duck.html {Copyrighted-reference only}
Posts: 41 | Registered: Tue October 28 2008 IP
Ignored post by PFF posted
Sat May 01 2010 04:22 PM
Belle lived in Choctaw Territory miles away. Besides, they had not seen each other in months."
Actually, Belle's cabin at Youngers Bend was in the Cherokee Nation--Just barely...
Quick Reply to: Who killed Belle Starr?

redpajama_set_name: RedPajamaCommonCrawl


text:
Q: Rails Strong Params, Accept Multiple Data Types Is there any way to allow Rails strong params to permit different data types? For example, I'm using react-bootstrap-typeahead and redux-form to create a form that allows users to select from provided values or create their own value. The provided values come from the database and are handed to the controller as an object with name and id whereas the user created values are passed as a string.

A: Permitting both a String and Array/Object in Rails 4+
TL;DR: Simply list parameter multiple times to support different types.

*

*String or Array: params.require(:my_model).permit(:my_attr, my_attr: [])

*String or Object: params.require(:my_model).permit(:my_attr, my_attr: [:key1, :key2])
Full Example
Suppose we have a model MyModel which has an attribute my_attr.
my_attr can be a string or array:
# example requests:
#   - {"my_model": {"my_attr": "hello" }
#   - {"my_model": {"my_attr": ["hello", "world"] }
params.require(:my_model).permit(:my_attr, my_attr: [])

my_attr can be a string or object:
# my_attr can be a string or an object with keys key1, key2
# example requests:
#   - {"my_model": {"my_attr": "hello" }
#   - {"my_model": {"my_attr": { "key1": "val1", "key2": "val2" }
params.require(:my_model).permit(:my_attr, my_attr: [:key1, :key2])

The first argument to permit allows my_attr to be a string, the second allows it to be an array or object (see documentation for Rails parameters).
Testing In Rails Console
Copy/Paste the following code to experiment with params directly:
# Testing with String
params = ActionController::Parameters.new(my_model: { my_attr: "My String" })
permitted = params.require(:my_model).permit(:my_attr, my_attr: [])
permitted.permitted?      # => true

# Testing with Array
params = ActionController::Parameters.new(my_model: { my_attr: ["Hello", "World"] })
permitted = params.require(:my_model).permit(:my_attr, my_attr: [])
permitted.permitted?      # => true

# Testing with Object
params = ActionController::Parameters.new(my_model: { my_attr: {key1: 'attr1', key2: 'attr2' }})
permitted = params.require(:my_model).permit(:my_attr, my_attr: [:key1, :key2])
permitted.permitted?      # => true

A: I recently did something like
params.require(:note).permit(
  fields: [:value, value: []]
)

which allowed me to submit string and array params.
Sorry I don't have the docs to back me up on that, I'm just learning ruby myself.  Hope this helps.

A: A form's submitted values will be stored in params[:object] regardless.  I'm still not sure what you're asking - can you post your create controller actions if that's what being called on submit?

redpajama_set_name: RedPajamaStackExchange


text:
Adventure, Family, Game, Gaming, Indie, Indie Dev, Music, Playstation, PlayStation 4, PS4, Rapid Reviews, Reviews, Rhythm
Mutazione Rapid Review
Title: Mutazione
Developer: Die Gute Fabrik
Publisher: Akupara Games
Website: https://mutazionegame.com/
Genre: Music, Rhythm, Adventure, Family
Price: £14.99 – Rapid Reviews was very kindly provided with a review code for this title.
For a game all about mutants, Mutazione might be one of the most human Indies I've played.
Mutazione places you in the running shoes of Kai, a girl who is tasked with sailing out to an island and looking after her ailing grandfather. Her grandfather is the shaman of an island full of mutated villagers, and he hopes to bond with his granddaughter and pass on some of his knowledge. Although the mutated villagers may seem like it'd be the focus of the story, the fact that they're mutated is secondary to their actual personalities which is a refreshing take.
It'll be pretty clear from the get-go that the presentation is a strong point of Mutazione. The art-style may not be complex, but it's unique and can be quite beautiful at times. Get your screenshot button ready. Although some may be put off by the lack of spoken dialogue, the game's soundtrack will be enough to guide you through. It's very peaceful and matches perfectly with the visuals and overall vibe of Mutazione.
Even with great presentation value, it's the characters that are going to keep you playing Mutazione. They are insanely well written and enjoyable to interact with, and it never feels like they only exist when Kai is around. You can tell all of the characters have their own relationship with one another, and it becomes pretty addicting checking in on them each day to see what's new.
Mutazione's writing surprised me on more than one occasion. One cool moment came when Kai insincerely gave another character a nickname, leading to them being open about how they use nicknames to try and form relationships with people more easily. It felt like the game had seen right through me. Another cool moment came when Kai was questioned about her love life, and she matter-of-factly talked about a girl she had a crush on back home. Once again, it felt like this was a world I was merely looking into, rather than one that had been created for me.
Once you arrive on the island, you'll be tasked with learning how to create and tend gardens, which is the game's primary gameplay mechanic besides talking to the other inhabitants of the island. It's not very complicated, but it serves as a way of driving the plot forward and giving the player more to do than walk and press a few buttons.
There are a lot of quality of life changes that would make Mutazione more enjoyable to play, although it's hard to fault the game since it's not always clear whether that's by design. Since you'll be spending most of your time running between areas to talk to people, it would be nice to have a way of getting around faster, but since the game is all about taking it slow, it seems like a purposeful choice to have you explore every time.
Mutazione offers a compelling story, full of interesting, likeable and incredibly real-feeling characters. Its central gameplay hook may not have grabbed me as its characters did, but it's still a fantastic Indie that's well worth a try.
You can purchase Mutazione from the PlayStation Store on the following link, https://store.playstation.com/en-gb/product/EP3245-CUSA14175_00-0000DGFMUTAZIONE
AdventureAkupara Games.Die Gute FabrikfamilyGameGamingIndieIndie DevMusicMutazionePlayStationPlayStation 4PS4Rapid ReviewsReviewsRhythmRRRRUK
By George Foster
Rush Rover Rapid Review
Gears Tactics Rapid Review
Cursed Castilla
Devious Dungeon 2
Dark Devotion Review


$463,829 VA Refinance in Canoga Park, CA. This was a first position loan with a Loan-to-Value of over 116%. James Lunnen was able to assist the borrower in lowering the interest rate on their mortgage. Their new interest rate is 3.750% with an APR of 3.742%. This loan was funded July 2012.


text:
Q: What extension is this? I was watching a programming video on YouTube and the instructor had the extension below installed. It looks very useful – instead of just showing open apps, it shows a wide range of default and customizable items.  Does anyone know what it is?

A: Looks like SuperTab ($20, free trial).


Of the 149,410 business administration professionals employed in New York, 3,700 work in Oswego. Between 2006 and 2010, in Oswego, there has been a 4% decline in the number of business administration professionals. Overall employment in Oswego has increased.
In Oswego, salaries for business administration professionals have increased. In 2010 Oswego business administration professionals earned an average yearly salary of $98,410. Business administration professionals in Oswego made a yearly average salary of $89,460, four years earlier in 2006. This growth is slower than the salary trend for all careers in Oswego.
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text:
+Lynda Kiejko
Lynda's Story
Dorothy Ludwig
Lynda Kiejko
Dorothy's Story
Dorothy started off her sports career in a variety areas such as basketball, badminton and even track and field prior to picking up a pistol for the first time. She always had athletic tendency and ability. Somewhere mixed in with a natural tendency to pick technique up quickly, and a passionate father, she found a love of shooting sports.
Dorothy was always the girl to beat in almost every competition. She always loved to compete. The 1995 Canada Winter Games was her first taste of a multi-sport games, and the sweet taste of victory. She not only was first place in the trials for these games, but she came home with a gold medal around her neck. From then on, nothing but the podium would due.
Even though training during her university years was difficult, she continued to train and stay on top of her game while achieving her Bachelor of Arts.
In 2002 Dorothy attended her first international multi-sport games, the Commonwealth Games in Manchester, England. Dorothy, ever mindful of walking in her father's footsteps was never one to be outdone, and not only matched her father, Bill Hare's 1978 Commonwealth Games Gold medal, but added a Silver medal to her tally as well. Gold and Silver medals in the Team, and Individual matches in Women's Air Pistol at the games. With her father Bill as her coach, and her brother John cheering her in the stands.
Dorothy continued to compete and strive for her very best. In 2005 her coach and father, passed away. She would have to spend the next years soul searching as to whether she would continue competing in her fathers memory, or if she was truly competing because she loved the sport.
Dorothy found that she couldn't walk away from the sport. And she was in it for her. She found her confidence and went into the 2007 Pan American games as Canada's stongest competitor and best chance for a medal. Through the finals in her event she went from first, to fourth. A Canadian Teammate captured the gold medal, and the trip to the Beijing 2008 Olympics. A devestating blow to any athlete to come so close, yet be so far away.
In 2010 Dorothy teamed up with her little sister Lynda at the Commonwealth games. The first time the two had been teammates internationally. Bringing home a bronze medal between them was a great reward. Even sweeter, was that Dorothy was pregnant and continued to compete during her pregnancy at two international competitions, both bringing home medals.
Facing motherhood, Dorothy was starting to think her sports career would be slowing down, however it was only just taking off.
Dorothy won the 2011 Canadian National Pistol Championships which named her to the 2011 Pan American Games team. Facing the Pan Am games a second time while also leaving her 6 month old son at home to compete was nothing less than a testement to her strength and resolve as a fierce competitor. Dorothy came home with the Gold Medal from the Pan American Games, and a spot on for the 2012 Olympics in London. All that stood between her and the Olympics now, was the Canadian team trials, where she would face off with her own sister.
Dorothy prepared herself, and came out victorious from the Canadian Trials! Without her sister pushing her to be her best, she was not sure she would have made it, so asked Lynda to join her Olympic Support Staff. A surreal experience to participate in the Olympics with her sister at her side. Her father would have been so proud!
Placing 34th in her first Olympics, and a strong performance, the sights are now set forward to what comes next. A quick turnaround brought Dorothy back to Canada to compete in the National Pistol Championships, solidifying once again her title as National Champion.
Through a whirlwind of post Olympic activity Dorothy welcomed her second child into her life in 2013. Training became much more challenging with two little ones to take care of, but she continued to find time to go to the range and put in the training of her much loved sport. True to herself and dedication Dorothy once again traveled abroad for the 2014 Commonwealth Games. Leaving two little boys behind, Dorothy and sister Lynda (with her 2 week old new baby in tow) over came many obstacles and barriers in order to remain a force to be reckoned with. Once again both sisters went into finals together and came out in 3rd and 5th. Dorothy was bringing home another Commonwealth medal (Bronze) to add to her collection.
That Dorothy's focus in University was for Drama and that for many years could be seen both on stage and in the technical booth of her local theater?
That one of Dorothy's favorite memories, is sitting beside her mom at a cabin, watching the sun rise over the mountains?
That standing next to her sister Lynda on the podium in 2010, was one of the best moments in her shooting career?
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