Early detection of wildlife morbidity and mortality through an event-based surveillance system.

The ability to rapidly detect and respond to wildlife morbidity and mortality events is critical for reducing threats to wildlife populations. Surveillance systems that use pre-diagnostic clinical data can contribute to the early detection of wildlife morbidities caused by a multitude of threats, including disease and anthropogenic disturbances. Here, we demonstrate proof of concept for use of a wildlife disease surveillance system, the 'Wildlife Morbidity and Mortality Event Alert System', that integrates pre-diagnostic clinical data in near real-time from a network of wildlife rehabilitation organizations, for early and enhanced detection of unusual wildlife morbidity and mortality events. The system classifies clinical pre-diagnostic data into relevant clinical classifications based on a natural language processing algorithm, generating alerts when more than the expected number of cases is recorded across the rehabilitation network. We demonstrated the effectiveness and efficiency of the system in alerting to events associated with both common and emerging diseases. Tapping into this readily available unconventional general surveillance data stream offers added value to existing wildlife disease surveillance programmes through a relatively efficient, low-cost strategy for the early detection of threats.

Evolutionary trajectory of SARS-CoV-2 and emerging variants.

The emergence of a novel coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and more recently, the independent evolution of multiple SARS-CoV-2 variants has generated renewed interest in virus evolution and cross-species transmission. While all known human coronaviruses (HCoVs) are speculated to have originated in animals, very little is known about their evolutionary history and factors that enable some CoVs to co-exist with humans as low pathogenic and endemic infections (HCoV-229E, HCoV-NL63, HCoV-OC43, HCoV-HKU1), while others, such as SARS-CoV, MERS-CoV and SARS-CoV-2 have evolved to cause severe disease. In this review, we highlight the origins of all known HCoVs and map positively selected for mutations within HCoV proteins to discuss the evolutionary trajectory of SARS-CoV-2. Furthermore, we discuss emerging mutations within SARS-CoV-2 and variants of concern (VOC), along with highlighting the demonstrated or speculated impact of these mutations on virus transmission, pathogenicity, and neutralization by natural or vaccine-mediated immunity.

Global shifts in mammalian population trends reveal key predictors of virus spillover risk.

Emerging infectious diseases in humans are frequently caused by pathogens originating from animal hosts, and zoonotic disease outbreaks present a major challenge to global health. To investigate drivers of virus spillover, we evaluated the number of viruses mammalian species have shared with humans. We discovered that the number of zoonotic viruses detected in mammalian species scales positively with global species abundance, suggesting that virus transmission risk has been highest from animal species that have increased in abundance and even expanded their range by adapting to human-dominated landscapes. Domesticated species, primates and bats were identified as having more zoonotic viruses than other species. Among threatened wildlife species, those with population reductions owing to exploitation and loss of habitat shared more viruses with humans. Exploitation of wildlife through hunting and trade facilitates close contact between wildlife and humans, and our findings provide further evidence that exploitation, as well as anthropogenic activities that have caused losses in wildlife habitat quality, have increased opportunities for animal-human interactions and facilitated zoonotic disease transmission. Our study provides new evidence for assessing spillover risk from mammalian species and highlights convergent processes whereby the causes of wildlife population declines have facilitated the transmission of animal viruses to humans.

Spread of Coxiella burnetii between dairy cattle herds in an enzootic region: modelling contributions of airborne transmission and trade.

Q fever, a worldwide zoonotic disease caused by Coxiella burnetii, is a looming concern for livestock and public health. Epidemiological features of inter-herd transmission of C. burnetii in cattle herds by wind and trade of cows are poorly understood. We present a novel dynamic spatial model describing the inter-herd regional spread of C. burnetii in dairy cattle herds, quantifying the ability of airborne transmission and animal trade in C. burnetii propagation in an enzootic region. Among all the new herd infections, 92% were attributed to airborne transmission and the rest to cattle trade. Infections acquired following airborne transmission were shown to cause relatively small and ephemeral intra-herd outbreaks. On the contrary, disease-free herds purchasing an infectious cow experienced significantly higher intra-herd prevalence. The results also indicated that, for short duration, both transmission routes were independent from each other without any synergistic effect. The model outputs applied to the Finistère department in western France showed satisfactory sensitivity (0.71) and specificity (0.80) in predicting herd infection statuses at the end of one year in a neighbourhood of 3 km around expected incident herds, when compared with data. The model developed here thus provides important insights into the spread of C. burnetii between dairy cattle herds and paves the way for implementation and assessment of control strategies.

A fresh look at the SarcoFluor antibody test for the detection of specific antibodies to Sarcocystis neurona for the diagnosis of equine protozoal myeloencephalitis.

Equine protozoal myeloencephalitis (EPM) is a challenging disease to diagnose in horses with neurological signs. To optimize contemporary diagnostic testing, including the use of serum:CSF antibody ratios, the SarcoFluor antibody test for Sarcocystis neurona requires revalidation. The SarcoFluor, a previously validated immunofluorescent antibody test (IFAT) for the detection of antibodies specific to S. neurona in serum and cerebrospinal fluid (CSF) of naturally infected horses was analyzed using recent data and considering a serum:CSF antibody ratio threshold. Utilization of serum and CSF phosphorylated neurofilament heavy protein (pNfH) concentrations in support of an EPM diagnosis was also evaluated. 172 horses were divided into three groups: EPM-positive horses (EPM+, n=42), neurological non-EPM horses (n=74) confirmed with non-EPM neurological diseases (cervical vertebral compressive myelopathy, equine neuroaxonal dystrophy/equine degenerative myeloencephalopathy), and control horses (control, n=56) without neurological signs and neurological abnormalities on histology. Logistic regression was used to compare EPM diagnostic regimens. Specifically, EPM+ horses were compared with neurological non-EPM horses showing neurological signs. To consider diagnostic utility, post-test probabilities were calculated by titer. When differentiating between EPM and other neurological diseases, the combination of serum and CSF SarcoFluor testing added more information to the model accuracy than either test alone. Using serum and CSF for pNfH in support of an EPM diagnosis did not identify cutoffs with statistically significant odds ratios but increased the overall model accuracy when used with the IFAT. Utilization of IFAT titers against S. neurona in serum and CSF result in a high post-test probability of detecting EPM+ horses in a clinical setting.

Conventional and frugal methods of estimating COVID-19-related excess deaths and undercount factors.

Across the world, the officially reported number of COVID-19 deaths is likely an undercount. Establishing true mortality is key to improving data transparency and strengthening public health systems to tackle future disease outbreaks. In this study, we estimated excess deaths during the COVID-19 pandemic in the Pune region of India. Excess deaths are defined as the number of additional deaths relative to those expected from pre-COVID-19-pandemic trends. We integrated data from: (a) epidemiological modeling using pre-pandemic all-cause mortality data, (b) discrepancies between media-reported death compensation claims and official reported mortality, and (c) the "wisdom of crowds" public surveying. Our results point to an estimated 14,770 excess deaths [95% CI 9820-22,790] in Pune from March 2020 to December 2021, of which 9093 were officially counted as COVID-19 deaths. We further calculated the undercount factor-the ratio of excess deaths to officially reported COVID-19 deaths. Our results point to an estimated undercount factor of 1.6 [95% CI 1.1-2.5]. Besides providing similar conclusions about excess deaths estimates across different methods, our study demonstrates the utility of frugal methods such as the analysis of death compensation claims and the wisdom of crowds in estimating excess mortality.

Red blood cell distribution width to platelet ratio in neonatal foals with sepsis.

BACKGROUND: Rapid and accurate markers to aid diagnosis of sepsis are needed in neonatal foals. The CBC variable red blood cell distribution width (RDW) to platelet ratio (RPR) is associated with inflammatory response and linked to poor outcomes of sepsis in human patients. HYPOTHESIS: Explore the correlation of RPR with sepsis in neonatal foals and evaluate RPR predictive and prognostic value. ANIMALS: Three hundred seventeen hospitalized neonatal foals ≤7 days of age that had a CBC and physical exam performed at admission between 2012 and 2021. METHODS: Retrospective case-control study. Clinical records were used to calculate sepsis scores and define groups. Red blood cell distribution width to platelet ratio was calculated and compared between groups (septic vs nonseptic) based on Kruskal-Wallis and Wilcoxon signed-rank tests. A multivariate logistic regression model to predict sepsis was created. The cutoff for RPR was obtained based on the maximal Youden Index. The Kaplan-Meier method and the log-rank test were used to estimate survival curves and compare survival rates based on RPR. RESULTS: Red blood cell distribution width to platelet ratio was significantly higher in septic foals (Median = 0.099, confidence interval [CI] [0.093; 0.108]) than in sick nonseptic (0.085, CI [0.083; 0.089]) and healthy foals (0.081, CI [0.077; 0.086]; P < .0001). Red blood cell distribution width to platelet ratio was able to predict sepsis with high accuracy (AUC = 82.1%). The optimal RPR cutoff for sepsis was 0.09. CONCLUSIONS AND CLINICAL IMPORTANCE: Red blood cell distribution width to platelet ratio calculation is practical, inexpensive, and based on CBC-derived data. Calculation of RPR along with CBC can aid in the diagnosis of sepsis and estimation of outcome.

Projecting the impact of an ebola virus outbreak on endangered mountain gorillas.

Ebola virus is highly lethal for great apes. Estimated mortality rates up to 98% have reduced the global gorilla population by approximately one-third. As mountain gorillas (Gorilla beringei beringei) are endangered, with just over 1000 individuals remaining in the world, an outbreak could decimate the population. Simulation modeling was used to evaluate the potential impact of an Ebola virus outbreak on the mountain gorilla population of the Virunga Massif. Findings indicate that estimated contact rates among gorilla groups are high enough to allow rapid spread of Ebola, with less than 20% of the population projected to survive at 100 days post-infection of just one gorilla. Despite increasing survival with vaccination, no modeled vaccination strategy prevented widespread infection. However, the model projected that survival rates greater than 50% could be achieved by vaccinating at least half the habituated gorillas within 3 weeks of the first infectious individual.

Evaluation of minimally invasive sampling methods for detecting Avipoxvirus: Hummingbirds as a case example.

Avian pox is a common avian virus that in its cutaneous form can cause characteristic lesions on a bird's dermal surfaces. Detection of avian pox in free-ranging birds historically relied on observations of visual lesions and/or histopathology, both which can underestimate avian pox prevalence. We compared traditional visual observation methods for avian pox with molecular methods that utilize minimally invasive samples (blood, toenail clipping, feathers, and dermal swabs) in an ecologically important group of birds, hummingbirds. Specifically, avian pox prevalence in several species of hummingbirds were examined across multiple locations using three different methods: (1) visual inspection of hummingbirds for pox-like lesions from a long-term banding data set, (2) qPCR assay of samples from hummingbird carcasses from wildlife rehabilitation centers, and (3) qPCR assay of samples from live-caught hummingbirds. A stark difference in prevalences among these three methods was identified, with an avian pox prevalence of 1.5% from banding data, 20.4% from hummingbird carcasses, and 32.5% from live-caught hummingbirds in California. This difference in detection rates underlines the necessity of a molecular method to survey for avian pox, and this study establishes one such method that could be applied to other wild bird species. Across all three methods, Anna's hummingbirds harbored significantly higher avian pox prevalence than other species examined, as did males compared with females and birds caught in Southern California compared with Northern California. After hatch-year hummingbirds also harbored higher avian pox prevalences than hatch-year hummingbirds in the California banding data set and the carcass data set. This is the first study to estimate the prevalence of avian pox in hummingbirds and address the ecology of this hummingbird-specific strain of avian pox virus, providing vital information to inform future studies on this charismatic and ecologically important group of birds.

Pathogen surveillance and epidemiology in endangered Peninsular bighorn sheep (Ovis canadensis nelsoni).

Peninsular bighorn sheep (Ovis canadensis nelsoni) are found exclusively in Southern California and Baja Mexico. They are federally endangered due to multiple threats, including introduced infectious disease. From 1981 - 2017, we conducted surveillance for 16 pathogens and estimated population sizes, adult survival, and lamb survival. We used mixed effects regression models to assess disease patterns at the individual and population levels. Pathogen infection/exposure prevalence varied both spatially and temporally. Our findings indicate that the primary predictor of individual pathogen infection/exposure was the region in which an animal was captured, implying that transmission is driven by local ecological or behavioral factors. Higher Mycoplasma ovipneumoniae seropositivity was associated with lower lamb survival, consistent with lambs having high rates of pneumonia-associated mortality, which may be slowing population recovery. There was no association between M. ovipneumoniae and adult survival. Adult survival was positively associated with population size and parainfluenza-3 virus seroprevalence in the same year, and orf virus seroprevalence in the previous year. Peninsular bighorn sheep are recovering from small population sizes in a habitat of environmental extremes, compounded by infectious disease. Our research can help inform future pathogen surveillance and population monitoring for the long-term conservation of this population.

Predicting the potential for zoonotic transmission and host associations for novel viruses.

Host-virus associations have co-evolved under ecological and evolutionary selection pressures that shape cross-species transmission and spillover to humans. Observed virus-host associations provide relevant context for newly discovered wildlife viruses to assess knowledge gaps in host-range and estimate pathways for potential human infection. Using models to predict virus-host networks, we predicted the likelihood of humans as hosts for 513 newly discovered viruses detected by large-scale wildlife surveillance at high-risk animal-human interfaces in Africa, Asia, and Latin America. Predictions indicated that novel coronaviruses are likely to infect a greater number of host species than viruses from other families. Our models further characterize novel viruses through prioritization scores and directly inform surveillance targets to identify host ranges for newly discovered viruses.

Tick infestation on wild snakes in northern part of western Ghats of India.

In total, 167 individuals of 30 species of snakes belonging to 22 genera and five families were examined for tick infestation from November 2008 to March 2010. Only two species of snakes, Ptyas mucosa (L., 1758) (Indian rat snake) and Naja naja (L., 1758) (spectacled cobra), were found infested by ticks. All ticks collected were identified to be Amblyomma gervaisi [previously Aponomma gervaisi (Lucas, 1847) 1. The average prevalence of these ticks on Indian rat snakes (n=48) was 29.16%, with abundance of 7.02 ticks per individual; on spectacled cobras (n=20), average prevalence was 30.00%, with abundance of 6.9 ticks per individual. The nymphs and males were predominant. All the ticks were found on the dorsal aspect of the body of the snake, and no ticks were recorded on the head, tail, or ventral body. The rate of tick infestation was highest in scrubland and was lowest in evergreen forests. Female Indian rat snakes showed higher tick infestation rates than male Indian rat snakes. Using Mann-Whitney U test, we found that longer snakes of both species had significantly higher rate of tick infestation in both the species of snakes.

Dairy management practices associated with multi-drug resistant fecal commensals and Salmonella in cull cows: a machine learning approach.

BACKGROUND: Understanding the effects of herd management practices on the prevalence of multidrug-resistant pathogenic Salmonella and commensals Enterococcus spp. and Escherichia coli in dairy cattle is key in reducing antibacterial resistant infections in humans originating from food animals. Our objective was to explore the herd and cow level features associated with the multi-drug resistant, and resistance phenotypes shared between Salmonella, E. coli and Enterococcus spp. using machine learning algorithms. METHODS: Randomly collected fecal samples from cull dairy cows from six dairy farms in central California were tested for multi-drug resistance phenotypes of Salmonella, E. coli and Enterococcus spp. Using data on herd management practices collected from a questionnaire, we built three machine learning algorithms (decision tree classifier, random forest, and gradient boosting decision trees) to predict the cows shedding multidrug-resistant Salmonella and commensal bacteria. RESULTS: The decision tree classifier identified rolling herd average milk production as an important feature for predicting fecal shedding of multi-drug resistance in Salmonella or commensal bacteria. The number of culled animals, monthly culling frequency and percentage, herd size, and proportion of Holstein cows in the herd were found to be influential herd characteristics predicting fecal shedding of multidrug-resistant phenotypes based on random forest models for Salmonella and commensal bacteria. Gradient boosting models showed that higher culling frequency and monthly culling percentages were associated with fecal shedding of multidrug resistant Salmonella or commensal bacteria. In contrast, an overall increase in the number of culled animals on a culling day showed a negative trend with classifying a cow as shedding multidrug-resistant bacteria. Increasing rolling herd average milk production and spring season were positively associated with fecal shedding of multidrug- resistant Salmonella. Only six individual cows were detected sharing tetracycline resistance phenotypes between Salmonella and either of the commensal bacteria. DISCUSSION: Percent culled and culling rate reflect the increase in culling over time adjusting for herd size and were associated with shedding multidrug resistant bacteria. In contrast, number culled was negatively associated with shedding multidrug resistant bacteria which may reflect producer decisions to prioritize the culling of otherwise healthy but low-producing cows based on milk or beef prices (with respect to dairy beef), amongst other factors. Using a data-driven suite of machine learning algorithms we identified generalizable and distant associations between antimicrobial resistance in Salmonella and fecal commensal bacteria, that can help develop a producer-friendly and data-informed risk assessment tool to reduce shedding of multidrug-resistant bacteria in cull dairy cows.

Retrospective study on admission trends of Californian hummingbirds found in urban habitats (1991-2016).

BACKGROUND: Hummingbirds are frequently presented to California wildlife rehabilitation centers for medical care, accounting for approximately 5% of overall admissions. Age, sex, and reason for admission could impact hummingbird survivability, therefore identification of these factors could help maximize rehabilitation efforts. METHODS: Mixed-effects logistic regression models were used to identify specific threats to the survival of 6908 hummingbirds (1645 nestlings and 5263 non-nestlings) consisting of five species (Calypte anna, Calypte costa, Selasphorus rufus, Selasphorus sasin, Archilochus alexandri), found in urban settings, and admitted to California wildlife rehabilitation centers over 26 years. RESULTS: In total, 36% of birds survived and were transferred to flight cage facilities for further rehabilitation and/or release. Nestlings were more likely to be transferred and/or released compared to adult hummingbirds. After accounting for age, birds rescued in spring and summer were twice as likely to be released compared to birds rescued in the fall. A high number of nestlings were presented to the rehabilitation centers during spring, which coincides with the nesting season for hummingbirds in California, with the lowest number of nestlings presented in fall. Reasons for presentation to rehabilitation centers included several anthropogenic factors such as window collisions (9.6%) and interactions with domesticated animals (12.9%). Survival odds were lower if a hummingbird was rescued in a "torpor-like state" and were higher if rescued for "nest-related" reasons. Evaluation of treatment regimens administered at wildlife rehabilitation centers identified supportive care, including providing commercial nutrient-rich nectar plus solution, to significantly increase hummingbird survivability. DISCUSSION: Our results provide evidence of threats to hummingbirds in urban habitats, based on reasons for rescue and presentation to rehabilitation centers. Reasons for hummingbird admissions to three California wildlife rehabilitation centers were anthropogenic in nature (i.e., being associated with domestic animals, window collisions, and found inside a man-made structure) and constituted 25% of total admissions. There was a clear indication that supportive care, such as feeding a commercial nectar solution, and medical treatment significantly increased the odds of survival for rescued hummingbirds.

TaqMan quantitative real-time PCR for detecting Avipoxvirus DNA in various sample types from hummingbirds.

BACKGROUND: Avian pox is a viral disease documented in a wide range of bird species. Disease-related detrimental effects can cause dyspnea and dysphagia, and birds with high metabolic requirements, such as hummingbirds, are thus especially vulnerable to the pathogen. Hummingbirds have a strong presence in California, especially in urban environments. However, little is understood regarding the impact of pox virus on hummingbird populations. Currently, diagnosing a pox infection relies on obtaining a tissue biopsy, which poses significant risks to birds and challenges in the field. Understanding the ecology of hummingbird pox viral infections could be advanced by a minimally invasive ante-mortem diagnostic method. Our aim was to address whether pox infections can be diagnosed using integumentary system samples besides tissue biopsies. To meet this goal, we tested multiple integumentary sample types using a quantitative real-time PCR assay. A secondary study goal was to determine which sample types (ranging from minimally to highly invasive sampling) were optimal for identifying infected birds. METHODOLOGY AND PRINCIPAL FINDINGS: Pox-like lesion tissue, pectoral muscle, feathers, toenail clippings, blood, and swabs (both pox-like lesion tissue and non pox-like lesion tissue) were taken from live birds and carcasses of two species of hummingbirds found in California. To maximize successful diagnosis, especially for samples with low viral load, a real-time quantitative PCR assay was developed for detecting the hummingbird-specific Avipoxvirus 4b core protein gene. Avipoxvirus DNA was successfully amplified from all sample types obtained from 27 individuals. These results were compared to those of conventional PCR and comparisons were also made among sample types, utilizing lesion tissue samples as the gold standard. CONCLUSIONS AND SIGNIFICANCE: Hummingbird avian pox can be diagnosed without relying on tissue biopsies. We identify that feather samples, of which contour feathers yielded the best results, can be used for diagnosing infected birds, thus reducing sampling risk. In sum, the real-time PCR assay detected viral DNA in various integumentary system sample types and will be useful in future studies of hummingbird disease ecology.

Vulnerabilities for Exposure to Emerging Infectious Disease at Urban Settlements in Nepal.

In Nepal, rapid urbanization and rural-to-urban migration especially due to internal civil conflict have catalyzed the development of temporary settlements, often along rivers on undeveloped land. This study conducted surveillance for viruses in small mammals and assessed potential risks for virus transmission to people in urban settlements along rivers in Kathmandu, Nepal. We collected samples from 411 small mammals (100 rodents and 311 shrews) at four riverside settlement sites and detected six viruses from four virus families including Thottapalayam virus; a strain of murine coronavirus; two new paramyxoviruses; and two new rhabdoviruses. Additionally, we conducted surveys of 264 residents to characterize animal-human contact. Forty-eight percent of individuals reported contact with wildlife, primarily with rodents and shrews (91%). Our findings confirm that rodents and shrews should be considered a health threat for residents of temporary settlements, and that assessment of disease transmission risk coupled with targeted surveillance for emerging pathogens could lead to improved disease control and health security for urban populations. Additionally, interventions focused on disease prevention should consider the unique urban ecology and social dynamics in temporary settlements, along with the importance of community engagement for identifying solutions that address specific multi-dimensional challenges that life on the urban river margins presents.

Correction: Use of RFID technology to characterize feeder visitations and contact network of hummingbirds in urban habitats.

[This corrects the article DOI: 10.1371/journal.pone.0208057.].

Predicting wildlife reservoirs and global vulnerability to zoonotic Flaviviruses.

Flaviviruses continue to cause globally relevant epidemics and have emerged or re-emerged in regions that were previously unaffected. Factors determining emergence of flaviviruses and continuing circulation in sylvatic cycles are incompletely understood. Here we identify potential sylvatic reservoirs of flaviviruses and characterize the macro-ecological traits common to known wildlife hosts to predict the risk of sylvatic flavivirus transmission among wildlife and identify regions that could be vulnerable to outbreaks. We evaluate variability in wildlife hosts for zoonotic flaviviruses and find that flaviviruses group together in distinct clusters with similar hosts. Models incorporating ecological and climatic variables as well as life history traits shared by flaviviruses predict new host species with similar host characteristics. The combination of vector distribution data with models for flavivirus hosts allows for prediction of  global vulnerability to flaviviruses and provides potential targets for disease surveillance in animals and humans.

Use of RFID technology to characterize feeder visitations and contact network of hummingbirds in urban habitats.

Despite the popular use of hummingbird feeders, there are limited studies evaluating the effects of congregation, sharing food resources and increased contact when hummingbirds visit feeders in urban landscapes. To evaluate behavioral interactions occurring at feeders, we tagged 230 individuals of two species, Anna's and Allen's Hummingbirds, with passive integrated transponder tags and recorded their visits with RFID transceivers at feeders. For detecting the presence of tagged birds, we developed an RFID equipped feeding station using a commercially available antenna and RFID transceiver. Data recorded included the number of feeder visits, time spent at the feeder, simultaneous feeder visitation by different individuals, and identifying which feeders were most commonly visited by tagged birds. For the study period (September 2016 to March 2018), 118,017 detections were recorded at seven feeding stations located at three California sites. The rate of tagged birds returning to RFID equipped feeders at least once was 61.3% (141/230 birds). Females stayed at feeders longer than males per visit. We identified primary, secondary and tertiary feeders at Sites 2 and 3, according to the frequency of visitation to them, with a mean percentage of 86.9% (SD±19.13) visits to a primary feeder for each tagged hummingbird. During spring and summer, hummingbirds visited feeders most often in morning and evening hours. Feeder visits by males overlapped in time with other males more frequently than other females. The analysis of the contact network at the feeders did not distinguish any significant differences between age or sex. Although most hummingbirds visited the feeders during the daytime, our system recorded night feeder visitations (n = 7 hummingbirds) at one site. This efficient use of RFID technology to characterize feeder visitations and contact networks of hummingbirds in urban habitats could be used in the future to elucidate behaviors, population dynamics and community structure of hummingbirds visiting feeders.