Quantifying animals' perception of environmental predictability using anticipatory behavior.

Animals under human care often experience predictable daily husbandry events, which can promote the development of anticipatory behavior. Previous research suggests even short delays in the arrival of a predictable, desired outcome can lead to negative welfare outcomes for animals. As such, providing reliable information to animals regarding the onset of important events may be a simple but useful method to support positive welfare. Here we evaluated the quantitative characteristics of anticipatory behavior of a California sea lion at the San Francisco Zoo in three situations (1) when the animal had accurate information about the occurrence of the event (temporally predictable training sessions), (2) when the information was semi-reliable (unscheduled training session), and (3) when a reliable signal was offered before unscheduled sessions. Results showed that providing a reliable cue resulted in a stronger temporal correlation of anticipation before the beginning of the unscheduled session, similar to the temporally predictable training session. However, providing a reliable cue did not reduce the intensity of the anticipatory behavior. We propose to take into account two aspects of the quantitative characteristics of anticipatory behavior: correlation with time until a desired event (correlation) and intensity, where the correlation indicates the ability of an individual animal to predict the occurrence of an event, and the intensity indicates the degree of sensitivity of the animal to reward. We discuss the implications for animal welfare and husbandry.

Keeper effect: Animals are more active in the presence of their caretakers.

Zookeepers are the primary caretakers of animals, providing daily care through frequent and close interactions. From the animal's perspective, most of these daily interactions are likely to have positive outcomes. With consistent and reliable interactions, a human-animal relationship is expected to develop. Our aim of this study was to investigate if the presence of the primary keeper in the public viewing area of zoo exhibits impacts the behavior of animals. We observed the behavior of 15 individuals of six species in the presence and absence of their primary keeper. Overall, we observed animals being more active in their keeper's presence than when the keeper was not present. When we considered if the keeper was nearby around times of offered opportunities to animals (e.g., feeding, enrichment, and training), our results showed that animals were equally as active when the keeper was present before an opportunity and when no opportunity was offered. These equal activity levels imply that the keeper is a cue for a forthcoming event to the animals, which reflects anticipatory behavior. Overall, we demonstrate that keeper presence is an environmental context in which animals behave differently than in keeper absence.

Dither: A unifying model of the effects of visitor numbers on zoo animal behavior.

Interest in the impact of human presence on the behavior and well-being of zoo and aquarium animals is increasing. Previous work has conceptualized the presence of zoo visitors as having one of three impacts on the behavior of animals in zoos: positive, negative, or neutral. Research suggests the same species may exhibit all three responses under different conditions, calling into question whether the positive/negative/neutral framework is the most useful way of considering visitor impact on animal behavior. Here we present a model of visitor effects that unifies these three predictions. Our model suggests that zoo-goers may provide a "dither effect" for some animals living in zoos. We posit animals may show nonlinear behavioral responses over a range of visitor densities, effectively exhibiting changes in both comfortable and anxiety-like behaviors under different levels of human presence. We tested this model during two COVID-19 related closures at the San Francisco Zoo, studying seven species for evidence of nonlinear relationships between visitor numbers and animal behavior. Our results support the dither effect acting in several species observed.

Host group formation decreases exposure to vector-borne disease: a field experiment in a 'hotspot' of West Nile virus transmission.

Animals can decrease their individual risk of predation by forming groups. The encounter-dilution hypothesis extends the potential benefits of gregariousness to biting insects and vector-borne disease by predicting that the per capita number of insect bites should decrease within larger host groups. Although vector-borne diseases are common and can exert strong selective pressures on hosts, there have been few tests of the encounter-dilution effect in natural systems. We conducted an experimental test of the encounter-dilution hypothesis using the American robin (Turdus migratorius), a common host species for the West Nile virus (WNV), a mosquito-borne pathogen. By using sentinel hosts (house sparrows, Passer domesticus) caged in naturally occurring communal roosts in the suburbs of Chicago, we assessed sentinel host risk of WNV exposure inside and outside of roosts. We also estimated per capita host exposure to infected vectors inside roosts and outside of roosts. Sentinel birds caged inside roosts seroconverted to WNV more slowly than those outside of roosts, suggesting that social groups decrease per capita exposure to infected mosquitoes. These results therefore support the encounter-dilution hypothesis in a vector-borne disease system. Our results suggest that disease-related selective pressures on sociality may depend on the mode of disease transmission.

A Quantitative Approach for Using Anticipatory Behavior as a Graded Welfare Assessment.

To perform quick assessments, welfare practitioners may focus on specific behavioral indicators of welfare, which can lead to challenges in interpretation. Anticipatory behavior has been suggested as a potentially graded indicator of well-being in animals. However, there are difficulties in assessing variations in this class of behavior quantitatively. Here, we propose an analytical approach for identifying and comparing the intensity of anticipatory behavior across different conditions. We evaluated the changes in the behavior of a sea lion at the San Francisco Zoo before and after daily training sessions, the start time of which had differing degrees of predictability. We show that anticipatory behavior is a complex suite of behaviors that can show multi-directional changes prior to an anticipated event. Additionally, we show that the methods utilized here can distinguish among differing intensities of anticipation directed toward daily husbandry events. We suggest that this approach may be broadly applicable for applying measures of anticipatory behavior as a graded welfare indicator.

Space, time, and context drive anticipatory behavior: Considerations for understanding the behavior of animals in human care.

Animal-based measures reflecting the welfare state of individuals are critical for ensuring the well-being of animals under human care. Anticipatory behavior is one potential animal-based measure that has gained traction in recent years, as it is theorized to relate to animals' reward sensitivity. It is of particular interest as an assessment for animals living under human care, as the predictability of the captive environment lends itself to the development of this class of behaviors. Animals are likely to exhibit anticipation in locations related to the anticipated event, often in temporally predictable time frames, and before specific contexts they experience in their day-to-day management. In this sense and under certain circumstances, anticipatory behaviors are likely to drive observed behavioral or space use patterns of animals under human care. Drawing conclusions from such data without identifying anticipation may result in misleading conclusions. Here we discuss how space, time, and context are related to patterns of anticipatory behaviors in animals under human care, how unidentified anticipation may alter conclusions regarding animal behavior or welfare under certain circumstances.

Assessing and Enhancing the Welfare of Animals with Equivocal and Reliable Cues.

The actions of human caretakers strongly influence animals living under human care. Here, we consider how intentional and unintentional signals provided by caretakers can inform our assessment of animals' well-being as well as help to support it. Our aim is to assist in further developing techniques to learn animals' affective state from their behavior and to provide simple suggestions for how animal caretakers' behavior can support animal welfare. We suggest that anticipatory behavior towards expected rewards is related to decision-making behavior as viewed through the cognitive bias lens. By considering the predictions of the theories associated with anticipatory behavior and cognitive bias, we propose to use specific cues to probe the cumulative affective state of animals. Additionally, our commentary draws on the logic of reward sensitivity and judgement bias theories to develop a framework that suggests how reliable and equivocal signals may influence animals' affective states. Application of this framework may be useful in supporting the welfare of animals in human care.

Managing Aged Animals in Zoos to Promote Positive Welfare: A Review and Future Directions.

Improvements in veterinary care, nutrition, and husbandry of animals living in zoos have led to an increase in the longevity of these animals over the past 30 years. In this same time period, the focus of animal welfare science has shifted from concerns over mitigating negative welfare impacts to promoting positive welfare experiences for animals. For instance, providing opportunities for animals to exert agency, solve problems, or acquire rewards are all associated with positive welfare outcomes. Many common age-related changes result in limitations to opportunities for positive welfare experiences, either due to pain or other physical, cognitive, or behavioral limitations. This review aggregates information regarding common age-related physical and behavioral changes across species, discusses how age-related changes may limit positive welfare opportunities of aged animals in human care, and suggests potential management methods to help promote positive welfare for animals at all life stages in zoos and aquariums.

Applying Behavioral Conditioning to Identify Anticipatory Behaviors.

The ability to predict regular events can be adaptive for nonhuman animals living in an otherwise unpredictable environment. Animals may exhibit behavioral changes preceding a predictable event; such changes reflect anticipatory behavior. Anticipatory behavior is broadly defined as a goal-directed increase in activity preceding a predictable event and can be useful for assessing well being in animals in captivity. Anticipation may look different in different animals, however, necessitating methods to generate and study anticipatory behaviors across species. This article includes a proposed method for generating and describing anticipatory behavior in zoos using behavioral conditioning. The article also includes discussion of case studies of the proposed method with 2 animals at the San Francisco Zoo: a silverback gorilla (Gorilla gorilla gorilla) and a red panda (Ailurus fulgens). The study evidence supports anticipation in both animals. As behavioral conditioning can be used with many animals, the proposed method provides a practical approach for using anticipatory behavior to assess animal well being in zoos.

Culex Flavivirus During West Nile Virus Epidemic and Interepidemic Years in Chicago, United States.

Culex flavivirus (CxFV) is an insect-specific flavivirus infecting Culex mosquitoes, which are important vectors of West Nile virus (WNV). CxFV and WNV cocirculate in nature and coinfect Culex mosquitoes, including in a WNV "hotspot" in suburban Chicago. We previously identified a positive association between CxFV and WNV in mosquito pools collected from suburban Chicago in 2006. To further investigate this phenomenon, we compared the spatial and temporal distribution of CxFV during an interepidemic year (2011) and an epidemic year (2012) for WNV. Both viruses were more prevalent in mosquito pools in 2012 compared to 2011. During both years, the CxFV infection status of mosquito pools was associated with environmental factors such as habitat type and precipitation frequency rather than coinfection with WNV. These results support the idea that WNV and CxFV are ecologically associated, perhaps because both viruses respond to similar environmental drivers of mosquito populations.

Crowdfunding Campaigns Help Researchers Launch Projects and Generate Outreach.

Organizers of participatory research (citizen science) projects can generate funds and outreach through crowdfunding. Here we provide insights from three successful science crowdfunding campaigns recently completed on Indiegogo, Experiment, and Kickstarter. Choosing a crowdfunding platform that fits the project is just the beginning; a successful campaign reflects its content, management, and marketing, and some researchers may need to acquire new skills. In addition, the growing trend of crowdfunding for science reinforces the importance of academic engagement with social media.

Effect of Trapping Methods, Weather, and Landscape on Estimates of the Culex Vector Mosquito Abundance.

The local abundance of Culex mosquitoes is a central factor adding to the risk of West Nile virus transmission, and vector abundance data influence public health decisions. This study evaluated differences in abundance estimates from mosquitoes trapped using two common methods: CO2-baited CDC light traps and infusion-baited gravid traps in suburban, Chicago, Illinois. On a weekly basis, the two methods were modestly correlated (r = 0.219) across 71 weeks over 4 years. Lagged weather conditions of up to four weeks were associated with the number of mosquitoes collected in light and gravid traps. Collections in light traps were higher with higher temperature in the same week, higher precipitation one, two, and four weeks before the week of trapping, and lower maximum average wind speed. Collections in gravid traps were higher with higher temperature in the same week and one week earlier, lower temperature four weeks earlier, and with higher precipitation two and four weeks earlier. Culex abundance estimates from light traps were significantly higher in semi-natural areas compared to residential areas, but abundance estimates from gravid traps did not vary by the landscape type. These results highlight the importance of the surveillance methods used in the assessment of local Culex abundance estimates. Measures of risk of exposure to West Nile virus should assess carefully how mosquito abundance has been estimated and integrated into assessments of transmission risk.

WEST NILE VIRUS ANTIBODY DECAY RATE IN FREE-RANGING BIRDS.

Antibody duration, following a humoral immune response to West Nile virus (WNV) infection, is poorly understood in free-ranging avian hosts. Quantifying antibody decay rate is important for interpreting serologic results and for understanding the potential for birds to serorevert and become susceptible again. We sampled free-ranging birds in Chicago, Illinois, US, from 2005 to 2011 and Atlanta, Georgia, US, from 2010 to 2012 to examine the dynamics of antibody decay following natural WNV infection. Using serial dilutions in a blocking enzyme-linked immunosorbent assay, we quantified WNV antibody titer in repeated blood samples from individual birds over time. We quantified a rate of antibody decay for 23 Northern Cardinals (Cardinalis cardinalis) of 0.198 natural log units per month and 24 individuals of other bird species of 0.178 natural log units per month. Our results suggest that juveniles had a higher rate of antibody decay than adults, which is consistent with nonlinear antibody decay at different times postexposure. Overall, most birds had undetectable titers 2 yr postexposure. Nonuniform WNV antibody decay rates in free-ranging birds underscore the need for cautious interpretation of avian serology results in the context of arbovirus surveillance and epidemiology.

Dispersal of adult culex mosquitoes in an urban west nile virus hotspot: a mark-capture study incorporating stable isotope enrichment of natural larval habitats.

Dispersal is a critical life history behavior for mosquitoes and is important for the spread of mosquito-borne disease. We implemented the first stable isotope mark-capture study to measure mosquito dispersal, focusing on Culex pipiens in southwest suburban Chicago, Illinois, a hotspot of West Nile virus (WNV) transmission. We enriched nine catch basins in 2010 and 2011 with 15N-potassium nitrate and detected dispersal of enriched adult females emerging from these catch basins using CDC light and gravid traps to distances as far as 3 km. We detected 12 isotopically enriched pools of mosquitoes out of 2,442 tested during the two years and calculated a mean dispersal distance of 1.15 km and maximum flight range of 2.48 km. According to a logistic distribution function, 90% of the female Culex mosquitoes stayed within 3 km of their larval habitat, which corresponds with the distance-limited genetic variation of WNV observed in this study region. This study provides new insights on the dispersal of the most important vector of WNV in the eastern United States and demonstrates the utility of stable isotope enrichment for studying the biology of mosquitoes in other disease systems.

An inverse association between West Nile virus serostatus and avian malaria infection status.

BACKGROUND: Various ecological and physiological mechanisms might influence the probability that two or more pathogens may simultaneously or sequentially infect a host individual. Concurrent infections can have important consequences for host condition and fitness, including elevated mortality risks. In addition, interactions between coinfecting pathogens may have important implications for transmission dynamics. METHODS: Here, we explore patterns of association between two common avian pathogens (West Nile virus and avian malaria parasites) among a suburban bird community in Chicago, IL, USA that share mosquito vectors. We surveyed 1714 individual birds across 13 species for both pathogens through established molecular protocols. RESULTS: Field investigations of haemosporidian and West Nile virus (WNV) infections among sampled birds yielded an inverse association between WNV serostatus and Plasmodium infection status. This relationship occurred in adult birds but not in juveniles. There was no evidence for a relationship between Haemoproteus infection and WNV serostatus. We detected similar prevalence of Plasmodium among birds captured with active WNV infections and spatiotemporally paired WNV-naïve individuals of the same species, demonstrating that the two pathogens can co-infect hosts. CONCLUSIONS: Mechanisms explaining the negative association between WNV serostatus and Plasmodium infection status remain unclear and must be resolved through experimental infection procedures. However, our results highlight potential interactions between two common avian pathogens that may influence their transmission among hosts. This is especially relevant considering that West Nile virus is a common zoonotic pathogen with public health implications. Moreover, both pathogens are instructive models in infectious disease ecology, and infection with either has fitness consequences for their avian hosts.

Prevalence of filarioid nematodes and trypanosomes in American robins and house sparrows, Chicago USA.

Hosts are commonly infected with a suite of parasites, and interactions among these parasites can affect the size, structure, and behavior of host-parasite communities. As an important step to understanding the significance of co-circulating parasites, we describe prevalence of co-circulating hemoparasites in two important avian amplification hosts for West Nile virus (WNV), the American robin (Turdus migratorius) and house sparrow (Passer domesticus), during the 2010-2011 in Chicago, Illinois, USA. Rates of nematode microfilariemia were 1.5% of the robins (n = 70) and 4.2% of the house sparrows (n = 72) collected during the day and 11.1% of the roosting robins (n = 63) and 0% of the house sparrows (n = 11) collected at night. Phylogenetic analysis of nucleotide sequences of the 18S rRNA and cytochrome oxidase subunit I (COI) genes from these parasites resolved two clades of filarioid nematodes. Microscopy revealed that 18.0% of American robins (n = 133) and 16.9% of house sparrows (n = 83) hosted trypanosomes in the blood. Phylogenetic analysis of nucleotide sequences from the 18s rRNA gene revealed that the trypanosomes fall within previously described avian trypanosome clades. These results document hemoparasites in the blood of WNV hosts in a center of endemic WNV transmission, suggesting a potential for direct or indirect interactions with the virus.