Marked spatiotemporal variabilities in mosquito infection of arboviruses, exemplified by the transmission of West Nile virus (WNV) in America, require adaptive strategies for mosquito sampling, pool screening and data analyses. Currently there is a lack of reliable and consistent measures of risk exposure, which may compromise comparison of surveillance data. Based on quantitative reasoning, we critically examined fundamental issues regarding mosquito sampling design and estimation of transmission intensity. Two surveillance strategies were proposed, each with a distinct focus, i.e. targeted surveillance for detection of low rates of mosquito infection and extensive surveillance for evaluation of risk exposure with high levels of mosquito infection. We strongly recommend the use of indicators embodying both mosquito abundance and infection rates as measures of risk exposure. Aggregation of surveillance data over long periods of time and across broad areas obscures patterns of focal arboviral transmission. We believe that these quantitative issues, once addressed by mosquito surveillance programs, can improve the epidemiological intelligence of arbovirus transmission.

The identification of the members of the Culex pipiens L. complex in arbovirus surveillance programs relies heavily on the use of morphology. In this work, we studied Cx. pipiens complex male mosquitoes collected from nine different locations, from northern, southern, and the hybrid zone sites in North America; Cairo, Egypt; and Nairobi, Kenya. Specimens were identified using DV/D ratio and also using amplification of the acetylcholinesterase (ACE.2) gene by both conventional and real-time polymerase chain reaction, and examination of the CQ11 locus. Consistent with previous findings, the morphological and molecular identifications did not always agree, particularly in regions of extensive introgression. There was an increased frequency of hybrid forms in late summer and early fall in Champaign Co., IL, that is north of the previously described Cx. pipiens complex hybrid zone. This represents an expansion of the North American hybrid zone. The biological and epidemiological relevance of the high degree of introgression and the late season increase in the proportion of intermediate forms is discussed.

In North America, West Nile and St. Louis encephalitis viruses have been detected in a wide range of vector species, but the majority of isolations continue to be from pools of mixed mosquitoes in the Culex subgenus Culex. Unfortunately, the morphologic identification of these important disease vectors is often difficult, particularly in regions of sympatry. We developed a sensitive real-time TaqMan polymerase chain reaction assay that allows reliable identification of Culex mosquitoes including Culex pipiens pipiens, Cx. p. quinquefasciatus, Cx. restuans, Cx. salinarius, Cx. nigripalpus, and Cx. tarsalis. Primers and fluorogenic probes specific to each species were designed based on sequences of the acetylcholinesterase gene (Ace2). Both immature and adult mosquitoes were successfully identified as individuals and as mixed species pools. This identification technique provides the basis for a rapid, sensitive, and high-throughput method for expounding the species-specific contribution of vectors to various phases of arbovirus transmission.

Of the 5,236 birds sampled for antibodies to West Nile virus (WNV) in Illinois from 2001 through 2004, 348 (6.6%) birds were seropositive. Our multiple year surveillance identified several avian species that had particularly high percentages of seropositive individuals. The importance of these species in the enzootic and/or epizootic transmission of WNV is discussed relative to their regional abundance and literature on host competency. The species with the highest exposure rates to WNV differed both temporally and regionally. In general, birds that bred or were born in Illinois were more likely to have antibodies than transient birds. There was also a significant difference in the seroprevalence between adults (12.1%) and juveniles (5.5%), indicating that the acquired antibody response from previous years is a critical concern when interpreting seroprevalence rates in wild-caught birds. The most common hosts for St. Louis encephalitis virus were also the most common hosts for WNV, which strongly supports the role of similar vectors for both flaviviruses. Avian species with high WNV seroprevalence rates tended to be those that bred throughout the year, have open cup nests, and live in close proximity to humans.

The relative efficacy of gravid and under-house CO2 traps for monitoring mosquito species of public health importance within the Houston metroplex area was assessed. Gravid and under-house traps were colocated at 10 sites and monitored weekly between 1 March to 31 May 2007. The most numerous species caught was Culex pipiens quinquefasciatus Say. Other species of public health importance caught in gravid and under-house traps included Culex restuans Theobald, Aedes aegypti (L.), and Aedes albopictus Skuse. Adjusting for the week of collection, gravid traps caught significantly more mosquitoes (mean 23.1 per trap) in the study area than under-house traps (mean 3.6 per trap). However, under-house traps caught a greater variety of mosquito species (13) than gravid traps (11). Gravid and under-house traps only caught nine of 15 of the same mosquito species during the study period. In this study area, gravid traps should be used as the primary method of surveillance for mosquito-borne diseases of public health importance during the early part of the season, because of greater catch numbers of mosquitoes that pose a public health risk.

We assessed the structure and latitudinal selection that might result in sensitivities to critical day-lengths that trigger diapause between Culex pipiens populations distributed along North-South and East-West axes in eastern North America. Strong population structure between Cx. p. pipiens and Cx. p. quinquefasciatus existed. Among Cx. p. pipiens, a 100-km increase in the latitudinal change resulted in an increased square root of F(ST) by 0.002. A 100-km increase in the longitudinal change caused an increased square root of F(ST) by 0.035. A lack of latitudinal influence on the structure between Cx. p. pipiens populations suggests a uniform signal using the 12 microsatellite markers, which might increase the risk of West Nile virus (WNV) transmission toward northern areas because of longer breeding season, extend host-seeking period, and larger population size. Northern Cx. p. pipiens may have undergone additional generations before diapause is triggered, magnifying population size when WNV amplification is peaking.

Australia experienced a large outbreak of equine influenza in August 2007. Nearly 10000 premises were infected during the epidemic. We used spatial and temporal analytical techniques to describe the epidemic, to quantify important descriptors of the epidemic, and to generate hypotheses about how the epidemic progressed and which control tools assisted in eradication. Spatio-temporal epidemic curves revealed three phases in the epidemic: dispersal, local spread and disease fade out. Spatial dispersal of infection rapidly declined immediately after national movement restrictions were introduced. The epidemic peak had passed before emergency vaccination could have induced substantial immunity in the equine population. Thirty seven clusters of epidemiologically linked premises were delineated using an interpolated surface of date of onset of clinical signs, geographic data and location of infected premises. These clusters were analysed individually to parameterise key epidemic measures: cumulative incidence, incidence rate, effective reproduction rate, nearest neighbour spread distances, epidemic length and the number of infected premises. These measures were summarised by landscape (rural versus peri-urban location) and standardised by disease management zone. Compared to rural areas, peri-urban areas appeared to have a higher density of equine premises (4.66 premises km(-2) versus 0.80 premises km(-2)), longer epidemics (95 days versus 87 days), more infected premises (393 versus 339) and a shorter spread distance (1.27km versus 2.38km). However, effective reproduction rates (2.04 versus 1.99), cumulative incidence (27.4 versus 26.9) and incidence rates (1.36 versus 1.54) were similar. The relative impact of vaccination and national movement restrictions in controlling this epidemic needs further investigation.

We investigated how the size and distribution of wild deer and feral pigs - species that might act as potential foot-and-mouth disease (FMD) virus maintenance hosts - might affect the persistence and spread of FMD. We used a susceptible-latent-infected-recovered geographic-automata model and spatially referenced data from southern Texas, USA. Within this study area, 100 locations were randomly selected and FMD virus spread was simulated (50 simulations each) at each location. As expected, the predicted sizes (km(2)) of the wild deer outbreaks were highly correlated (r(SP)>0.95) with the number of deer at incursion locations, the total number of deer within 2km of incursion locations, and the minimum and maximum deer herd size within 2km of incursion locations. However, the predicted sizes of the feral pig outbreaks were only moderately correlated (r(SP) 0.63-0.67) with the total, maximum and variance of the number of feral pigs within 2km of incursion locations. Lack of continuity within the feral pig herd distribution across the landscape makes predicting disease spread more difficult than for deer, a more homogenously distributed species. When assessing the potential of wild and feral animal species at a locality to act as maintenance hosts of FMD virus, estimates of the population size and distribution might serve as a useful indicator of potential outbreaks in some circumstances.

Soon after the appearance of West Nile virus (WNV) in North America, a number of public health authorities designated the American Crow (Corvus brachyrhynchos) a sentinel for WNV detection. Although preliminary studies have suggested a positive association between American Crow mortality and increased risk of WNV infection in humans, we still know little about dynamic variation in American Crow mortality, both baseline levels and mortality associated with WNV. We hypothesized that the complex social behavior of American Crows, which is shaped by age and seasonal factors, influences both baseline mortality and WNV mortality in American Crow populations. We examined American Crow mortality data from Quebec for the 2005 WNV surveillance year, which lasted from 5 June to 17 September 2005. The variables of interest were age, gender, body condition index, time of year, and land cover. We used a log-linear model to examine baseline mortality. Logistic regression and general linear regression models were constructed to examine variables associated with mortality due to WNV. We found that both age and time of year were key variables in explaining baseline mortality. These two variables were also risk factors for WNV mortality. The probability that a carcass tested positive for WNV increased with the age of the dead bird and as summer progressed. WNV-positive carcasses also had a lower body condition index than WNV-negative carcasses. We believe that the first major wave of American Crow mortality observed in the early summer of 2005 was the result of natural mortality among young American Crows. Because this mortality was not linked to WNV, it appears that American Crow may not be a good species for early detection of WNV activity. Our data also suggest that second-year American Crows play a major role in propagating WNV during their movements to urban land covers during midsummer.

New Caledonian crows Corvus moneduloides use tools made from sticks or leaf stems to 'fish' woodboring beetle larvae from their burrows in decaying wood. Previous research on this behaviour has been confined to baited sites, leaving its ecological context and significance virtually unexplored. To obtain detailed observations of natural, undisturbed tool use, we deployed motion-triggered video cameras at seven larva-fishing sites. From 1797 camera hours of surveillance over 111 days, we recorded 317 site visits by at least 14 individual crows. Tool use was observed during 150 site visits. Our video footage revealed notable variation in foraging success among identifiable crows. Two nutritionally independent, immature crows spent considerable time using tools, but were much less successful than local adults, highlighting the potential role of individual and social learning in the acquisition of tool-use proficiency. During systematic surveys of larva-fishing sites, we collected 193 tools that crows had left inserted in larva burrows. Comparing these tools with the holes in which they were found, and with raw materials available around logs, provides evidence for tool selectivity by New Caledonian crows under natural conditions. Taken together, these two complementary lines of investigation provide, to our knowledge, the first quantitative description of larva fishing by wild crows in its full ecological context.

Summary After 10 years, West Nile virus (WNV) re-emerged in Italy in August 2008. As on 31 December 2008, the infection affected eight Provinces in three Regions (Emilia Romagna, Veneto, Lombardy), where a total of 794 cases of WNV infection in 251 equine stables were detected on the basis of the clinical signs and as a result of a serological screening in horses living in the area. Only 4.0%(32/794) of the serologically positive animals showed clinical signs, and the 32 clinical cases were reported in 18 different farms. The observed case-fatality rate was 15.6%(5/32). The confirmed clinical cases were detected from end August to mid October. Significant levels of positivity by RT-PCR were also observed in magpies (Pica pica)(9.1%, 95% confidence levels: 6.1-13.4%), carrion crows (Corvus corone)(7.4%, 95% confidence levels: 3.6-14.4%) and rock pigeons (Columba livia)(12.9%, 95% confidence levels: 7.6-21.2%).

Creative problem solving and innovative tool use in animals are often seen as indicators of advanced intelligence because they seem to imply causal reasoning abilities [1]. However, complex behavior can also arise from relatively simple mechanisms [2, 3], and the cognitive operations underlying seemingly "insightful" behavior are rarely examined [4]. By controlling and varying prior experience, it is possible to determine the minimum information animals require to solve a given problem [5]. We investigated how pretesting experience affects the performance of New Caledonian crows (Corvus moneduloides) when facing a novel problem. The task (developed by Bird and Emery [6]) required dropping stones into a vertical tube to collapse an out-of-reach platform in a transparent box and release a food reward. After establishing that the birds had no preexisting tendency to drop stones into holes, subjects were assigned to two experimental groups that were given different kinds of experience with the affordances of the apparatus. Crows that had learned about the mechanism (collapsibility) of the platform without the use of stones passed the task, just like the subjects that had previously been trained to drop stones. This demonstrates that successful innovation was also possible after acquaintance with just the functional properties of the task.

The American crow (Corvus brachyrhynchos) is known to suffer 100% mortality from infection with the New York 1999 strain of West Nile virus (WNV). Following the initial detection of WNV in North America in 1999, we measured prevalence of WNV-reactive antibodies ("seroprevalence") in free-ranging American and fish crows (Corvus ossifragus) of central New Jersey after each transmission season through 2005. In 2002, seroprevalence in American crow juveniles increased to 14% from the 5% of the previous year, potentially indicating increased survival in this species. Using the annual seroprevalence measurements and the number of human West Nile neuroinvasive disease cases as a surrogate for WNV transmission intensity, we developed a model to estimate the annual WNV-associated mortality rates among both of these crow species. Our model supports the hypothesis that mortality is changing over time; the WNV-associated mortality rate declined over time by 1.5% for American crow and by 1.1% for fish crow. The probability that the trend in mortality was negative was 90% for the American crow and 60% for the fish crow.

Summary West Nile virus (WNV) infection, a zoonotic disease for which birds act as a reservoir, first appeared in North America in August 1999. It was first reported in Quebec in 2002. The Quebec surveillance system for WNV has several components, including the surveillance of mortality in corvid populations, which includes the American crow (Corvus brachyrhynchos). The main objectives of this study are to better understand the population dynamics of this species in Quebec and to evaluate the impact of WNV on these dynamics. We obtained observation data for living crows in this province for the period of 1990-2005 and then conducted a spectral analysis of these data. To study changes in crow population dynamics, the analysis was carried out before and after the appearance of WNV and space was divided in two different areas (urban and non-urban). Our results show the importance of cycles with periods of less than 1 year in non-urban areas and cycles with periods of greater than 1 year in urban areas in the normal population dynamics of the species. We obtained expected fluctuations in bird densities using an algorithm derived from spectral decomposition. When we compared these predictions with data observed after 2002, we found marked perturbations in population dynamics beginning in 2003 and lasting up to 2005. In the discussion, we present various hypotheses based on the behaviour of the American crow to explain the normal population dynamics observed in this species and the effect of type of area (urban versus non-urban). We also discuss how the predictive algorithm could be used as a disease surveillance tool and as a measure of the impact of a disease on wild fauna.

Abstract Birds are usually considered beneficial partners for plants, acting as predators on herbivorous insects, pollinators and seed dispersal agents. However, in an urban area of central Japan, birds break off large quantities of live camphor tree (Cinnamomum camphora) twigs in winter. This loss of vegetative parts was examined quantitatively to estimate the impact on the trees. I also observed bird foraging behaviour to determine the species involved and the possible reasons underlying this destructive activity. Broken twigs on the forest floor were found to have numerous leaves and spring buds. The densities of leaves and buds in the litter were 288.5 and 54.4 m(-2), respectively. The jungle crow (Corvus macrorhynchos) may have broken off the twigs either to peck the fruits while perching on stable branches, or possibly to remove twigs obstructing access to fruit. In contrast, brown-eared bulbuls (Hypsipetes amaurotis), oriental turtle doves (Streptopelia orientalis) and rove doves (Columba livia) ate fruits without breaking twigs. The interaction between C. camphora and C. macrorhynchos only extends back for about 20 years in urban Japan, indicating that this is unlikely to be a stable, co-evolved relationship.

A new original method to investigate some cognitive abilities of wild passerine birds was developed. In the course of experiments with five crossbills (Loxia curvirostra), it was shown that the method can be used to evaluate the ability of birds to form concepts. Crossbills were presented with pairs of stimuli each consisting of 1 to 10 equal elements. All the five crossbills learned to choose, within a pair, a stimulus with a larger array of elements. The birds successfully transferred this performance to novel stimuli considerably different from the training pairs comparable with the latter only in the number or area. One of the birds showed a complete transfer of the acquired performance of choice to stimuli consisting of 10 to 20 equal elements. Thus, the ability of forest passerine birds to form concept "larger than" was shown.

After the 2001 occurrence of West Nile virus (WNV) in Wisconsin (WI), we collected sera, during 2003-2006, from south-central WI mesopredators. We tested these sera to determine WNV antibody prevalence and geometric mean antibody titer (GMAT). Four-fold higher antibody prevalence and 2-fold higher GMAT in 2003-2004 indicated greater exposure of mesopredators to WNV during the apparent epizootic phase. The period 2005-2006 was likely the enzootic phase because WNV antibody prevalence fell to a level similar to other flaviviruses. Our results suggest that, in mesopredators, vector-borne transmission is the primary route of infection and WNV antibodies persist for < 1 year. Mesopredators may be sensitive indicators of West Nile virus spill-over into humans and horses. Mesopredator sero-surveys may complement dead crow surveillance by providing additional data for the timing of public health interventions. Research is needed to clarify the dynamics of WNV infection in these mammals and their role as potential WNV amplifiers.

The use of roosting sites by animal societies is important in conservation biology, animal behavior, and epidemiology. The giant noctule bat (Nyctalus lasiopterus) constitutes fission-fusion societies whose members spread every day in multiple trees for shelter. To assess how the pattern of roosting use determines the potential for information exchange or disease spreading, we applied the framework of complex networks. We found a social and spatial segregation of the population in well-defined modules or compartments, formed by groups of bats sharing the same trees. Inside each module, we revealed an asymmetric use of trees by bats representative of a nested pattern. By applying a simple epidemiological model, we show that there is a strong correlation between network structure and the rate and shape of infection dynamics. This modular structure slows down the spread of diseases and the exchange of information through the entire network. The implication for management is complex, affecting differently the cohesion inside and among colonies and the transmission of parasites and diseases. Network analysis can hence be applied to quantifying the conservation status of individual trees used by species depending on hollows for shelter.