Deer Populations Research Articles

Possibility of Vertical Transmission of Sarcocystis Spp. in Sika Deer in Japan.

In recent years, the wild deer population in Japan has grown exponentially, causing severe feeding damage to the agricultural and forestry industries. Therefore, the game meat industry is being promoted for effective utilization of hunted animals. Wild animals are not hygienically controlled and can serve as reservoirs for pathogenic microorganisms. However, epidemiological information on wild animals in Japan remains insufficient. Recently, food poisoning-like cases have occurred because of raw venison infection with Sarcocystis spp. As the prevalence of Sarcocystis spp. in sika deer is very high in Japan and even fawns are infected, this study attempted to verify the vertical infection of Sarcocystis spp. in sika deer in Japan. Genetic detection of Sarcocystis 18S ribosomal RNA in fetal and maternal tissues from early to late gestation in sika deer revealed Sarcocystis Types 1-5 and Sarcocystis fayeri in the mother and fetus. Types 1, 2, 4, and 5 were detected in the maternal tissues of Ezo sika deer (Cervus nippon yesoensis) in Hokkaido, whereas Types 1 and 2 and S. fayeri were detected in fetuses. Types 1-5 were detected in Honshu sika deer (Cervus nippon centralis) in Mie Prefecture but not in the fetuses. Types 1, 2, and 4 were detected in the udder and milk samples. This indicates that Sarcocystis Types 1 and 2 and S. fayeri have the ability to pass through the placenta of sika deer and invade fetal tissues and Types 1, 2, and 4 may be transmitted orally via milk. These findings suggest that there is transplacental and transmammary transmission of Sarcocystis spp. in sika deer.

Identification of Factors Associated with Both Frequency and Severity of Deer-Related Motorcycle Crashes.

Motorcyclists are particularly vulnerable to injury when encountering large animals such as deer in the roadway. The frequency of these events, factors associated with their occurrence, and the extent of injuries sustained from such crashes are not yet well described. A review of 10 years (2012-2021) of statewide motorcycle crash reports from a single state with a large deer population was performed to ascertain the frequency of deer-associated motorcycle crashes. Environmental and roadway conditions, as well as motorcyclist age, gender, and helmet usage, were identified to determine if associations exist with the severity of injuries sustained. Statewide, 8.5% of reported motorcycle crashes involved deer. Excluding the two most densely populated counties, this rose to 10.7%, with 24.1% of crashes occurring during non-daylight hours being deer related. Factors associated with increased frequency of more severe injury and death included not wearing a helmet, female gender, and rural county crash location. Death and severe injuries were reported in more than twice as many riders who were unhelmeted compared to those wearing helmets (36.1 vs 16.6%; p < 0.01). In a single midwestern state, deer represent a significant danger to motorcyclists, especially in rural settings during non-daylight hours. A cautious approach to riding during this time, utilizing available safety precautions to minimize risk of severe injury and death, and further exploration of viable interventions to reduce the likelihood of these occurrences should be considered.

Cascading effects driven by population recovery of sika deer on habitat use of sympatric mammals under heavy snow conditions

AbstractUnlike the top‐down cascading effects driven by apex predators, the bottom‐up effects on biological interactions originating from herbivory by large mammals, especially within mammalian assemblages, have received less attention. This study aimed to identify the multifaceted impacts driven by sika deer (Cervus nippon) during midwinters, when their herbivory impacts were expected to be strong due to deficient dietary resources. We focused on interference and exploitative competition caused by deer on the habitat use of four sympatric mammal species, namely two herbivores (Japanese serow, Capricornis crispus, and Japanese hare, Lepus brachyurus angustidens) and two mesocarnivores (red fox, Vulpes vulpes, and Japanese marten, Martes melampus melampus). To estimate the wintering behaviors of these mammals and deer herbivory pressure, we performed snow tracking and counted the fresh feeding marks of deer during midwinter in 2022 and 2023. We conducted surveys on 201‐km transects with different stages of deer population recovery (i.e., different deer population densities) in heavy snowfall regions of Japan. We then systematically quantified the deer‐driven effects on sympatric mammals using piecewise structural equation modeling (SEM). We recorded 1700 tracks of target mammals and 1327 trees with deer feeding marks. Based on these records, we constructed an SEM with acceptable performance (Fisher's C = 27.1, P = 0.30). The SEM suggested some possibilities that exploitative competition mediated by deer herbivory constrained the habitat use of mammals with dietary habits similar to that of deer, i.e., serow (effect size, −0.13) and hares (−0.14). In addition, through constricting the habitat use of hares, the deer‐driven cascading effects reduced the occurrence of their key predators, i.e., foxes (−0.12) and martens (−0.10). Thus, our observations provide novel evidence that food webs are regulated by herbivore‐driven bottom‐up cascading effects in cases where plant primary productivity is limited by heavy snowfall.

Ecosystem transition due to deer overabundance: Insights from long‐term studies and future considerations

AbstractThe overabundance of certain deer species is emerging as a critical issue in many forested regions across the Northern Hemisphere, including those in Japan. In the field of deer impact studies, the prevailing notion has been that overabundant deer populations could cause drastic changes of ecosystem states. This study comprises a review of the historical discussion surrounding ecosystem changes caused by deer overabundance, from theoretical frameworks to in situ observations and experiments. The synthesis highlights the potential for state transitions, shifting ecosystems from forest to nonforest states, and in some scenarios to a so‐called alternative stable states. However, detecting these transitions poses challenges due to the enduring impacts of past deer activity and the nonequilibrium nature of forest dynamics. Furthermore, this study reveals additional multifield complexities arising from the interactive effects of deer overabundance and global changes on future forest dynamics. To address these challenges, new avenues for research are proposed, emphasizing the importance of sustained efforts in conducting valuable long‐term studies.

Wolf Predation on White‐tailed Deer Before, During, and After a Historically Mild Winter in Northern Minnesota

ABSTRACTIn many southern boreal ecosystems of North America, wolves are the primary predators of white‐tailed deer, and white‐tailed deer are the primary prey of wolves. Furthermore, wolf–deer systems have and will continue to become more common as white‐tailed deer range continues expanding northward in North America. Despite this, there is little information on kill rates of wolves on deer (i.e., the number of deer killed per wolf per unit of time)—a fundamental metric of wolf predation on deer—and how kill rates vary with deer density, wolf density, and environmental conditions. We estimated kill rates of wolves on deer before, during, and after a historically mild winter in the Greater Voyageurs Ecosystem, Minnesota, USA. Kill rates of wolves on deer were low (0.009–0.018 deer/wolf/day) in fall, peaked in February (0.050 deer/wolf/day), and quickly declined to 0 deer/wolf/day by April. The kill rates of wolves on deer we observed in winter were some of the lowest kill rates of wolves on deer that have been documented. Wolves in the Greater Voyageurs Ecosystem appeared unable to catch and kill a sufficient number of deer to meet their daily energetic requirements during Winter 2023–2024, and thus most wolves likely lost weight during winter, a period when wolves are typically in peak physical condition. The rates of wolf predation we observed appeared to be well below those needed to decrease deer population density in the GVE. Thus, our work, in combination with numerous other studies, indicates winter conditions are the primary driver of deer population change in northern climates.

Molecular characteristics of CTX-M β-lactamase-producing and quinolone-resistant Escherichia coli among deer in a popular tourist spot in Japan.

Introduction. Antimicrobial resistance (AMR) is a growing global concern. Clonal lineages of CTX-M β-lactamase-producing Escherichia coli (CTXE) and quinolone-resistant E. coli (QREC) were disseminated among the deer population in a famous tourist destination (Nara Park; NP) in Japan. Hypothesis/gap statement. The molecular characteristics of CTXE or QREC isolates, which could pose a threat to public health, have not been elucidated. Aim. This study aimed to characterize the genetic traits of CTXE and QREC isolates derived from NP deer and compare them with lineages prevalent worldwide. Methodology. Sixteen CTXE and three QREC isolates recovered from NP deer faeces between 2018 and 2020 were analysed using whole-genome sequencing (WGS). For endemic lineages, phylogenetic trees were constructed against the isolates registered in the EnteroBase database using the core genome SNP scheme. Results. The most prevalent lineage in NP deer was ST3580. Several pandemic lineages, such as sequence type (ST) 38, ST58 and ST117, were included. The QREC lineages prevalent among deer were designated as extra-intestinal pathogenic E. coli or uropathogenic E. coli (UPEC). Thirteen of the 24 antimicrobial resistance genes (ARGs) were considered high-risk ARG families. Chromosomal integration of bla CTX-M-15 was observed in all plasmid-negative isolates. Phylogenetic analysis suggested relationships between NP isolates and isolates sourced from the environment or poultry. Conclusion. ST3580 has a high potential for clonal dissemination. Furthermore, multiple clinically relevant lineages of CTXE and QREC are endemic in NP deer; however, they could be less virulent than isolates belonging to the same lineages, which could cause severe infectious diseases. Further studies are required to investigate the relationship between chromosomal integration of plasmid-encoded genes and the stable propagation of AMR bacteria in wildlife and the environment.

An Updated Framework for Modeling White-Tailed Deer (Odocoileus virginianus) Habitat Quality in Illinois, USA.

White-tailed deer (Odocoileus virginianus) are a cervid species found mostly in the Americas. Managing white-tailed deer requires understanding their relationship with the environment, which was characterized by Roseberry and Woolf (Wildlife Society Bulletin 1, 1998, 252) for all counties in Illinois, USA, who incorporated habitat quantity and quality in a deer habitat suitability index. However, this index was based on satellite imagery from 1996 and did not explore the smaller spatial scales used by deer. Our study addressed these gaps by developing a deer land cover utility (LCU) score for each TRS (township, range, and section), township, and county in Illinois based on the methodology outlined in Roseberry and Woolf (Wildlife Society Bulletin 1, 1998, 252) but using data from the National Land Cover Database (2001-2021). These deer LCU scores were validated against minimum deer population data using Bayesian regression with additional covariates relevant to hunting and deer density. These models performed well with Bayesian R 2 values of 0.501 (TRS), 0.5 (township), and 0.969 (county). The regression coefficients for the deer LCU scores were statistically significant (95% credibility interval not containing 0) and positive at the TRS, township, and county levels, reflecting the expected relationship between minimum deer density and deer LCU. Predictions made by these regression models on new data were accurate, with the median absolute difference between the true and predicted values being 0.398 deer/km2 for TRS', 0.085 deer/km2 for townships, and 0.066 deer/km2 for counties. This deer LCU could be used in other studies about deer in Illinois or studies in which deer are a relevant factor such as investigations about deer disease or tick distribution. This modeling approach could also be adapted to different wild species, locations, and/or time periods for which land cover data is available.

Immunocastration caused dysfunction of Père David’s deer (Elaphurus davidianus)

Abstract The population density of Père David’s deer (Elaphurus davidianus) in Jiangsu Dafeng Milu Nature Reserve was excessively high due to an overemphasis on population recovery, while ignoring the crucial relationship between population size and habitats. This study used immunocastration to capture four Père David’s deer and observed their behavior. Their blood was collected for sex hormone testing, transcriptomics, and metabolomics analysis. In addition, six uncastrated Père David’s deer were included as control group. The results showed that reproductive behavior was absent in castrated Père David’s deer, while castrated individuals exhibited a significant decrease in blood hormone testosterone concentration compared to uncastrated Père David’s deer. Transcriptome and metabolome analyses revealed 450 differentially expressed genes (DEGs) and five differentially expressed metabolites (DEMs), Combined analysis of transcriptome and metabolic data revealed a significant correlation between DEGs and DEMs. It shows that using immune castration to control the population of Père David’s deer may disrupt their cell apoptosis and disturb the cell cycle, leading to abnormal pathways and promoting the onset of many diseases. Therefore, a cautious approach should be adopted toward immune castration as a means of population control. This study can provide valuable references for the standardized management of Père David’s deer populations.

Facts about Wildlife Diseases: Warts in White-tailed deer (Cutaneous Fibromas).

Deer warts, or cutaneous fibromas, are skin tumors caused by papillomavirus infection that affect white-tailed deer and other deer. These growths are generally benign, but in rare cases may proliferate and impede functions like sight, movement, or feeding. Deer immune systems usually eradicate the infection over time. Found exclusively on the skin, usually around the eyes, neck, face, and forelegs, warts may be diverse in appearance and size, ranging from small fleshy nodules to larger, pendulous masses or clusters. Transmission mechanisms are suspected to involve skin contact or possibly insect vectors but remain unclear. Diagnosis is by visual examination and histopathological analysis of the wart tissue. Wild deer populations usually do not require treatment, but surgical removal is an option for captive herds. Vaccine development remains plausible but has not yet been pursued extensively.

Variation in white‐tailed and mule deer dental metrics is associated with precipitation gradients in the Americas

AbstractInvestigating the relationship between the physical environment and organismal adaptation is crucial to understanding the selective pressures that drive mammalian evolution. Of the varied approaches to exploring this relationship, macroscopic tooth wear analyses have become widely used due to their economic and simple methodologies. However, more work is needed to understand how assessments of wear patterns operate within finer levels of dietary and environmental contexts. It is specifically unclear if and how tooth wear differs inter‐ and intraspecifically between geographically widespread, large‐bodied mammalian populations. This project characterizes molar hypsodonty indices and mesowear scores of white‐tailed deer (Odocoileus virginianus) and mule deer (Odocoileus hemionus) populations across North and Central America, investigating if and how molar wear and morphology vary over the deer's large and ecologically variable geographic distribution. While mesowear scores generally do not strongly track environmental signals within and between Odocoileus species, hypsodonty indices reveal a strong correlation between crown height and aridity, which likely represents an adaptive response to abrasive diets in areas with low precipitation.

Roe Deer Reproduction in Western Poland: The Late Autumn Rut Phenomenon.

Roe deer (Capreolus capreolus L.) populations in Poland are characterized by low productivity, which is why their reproductive potential was investigated. The presence of corpora lutea (CL) on the ovaries of females hunted in autumn and winter in the years 2015 and 2016 was assessed. Most animals were post-rut and most often had multiple ovulations. However, in early autumn 2015, 60% of the females had no CL. Therefore, they did not participate in mating at the turn of July/August. Those that did ovulate were found to have one CL. In late autumn, 97% of the females were post-rut, mostly with twin ovulations. This confirmed the occurrence of a late autumn rut. This phenomenon had been suggested in studies but not confirmed. In contrast, almost 100% of roe deer ovulated in summer in 2016. It was hypothesized that the reason for the low mating activity of roe deer in the summer of 2015 was heat stress and limited food resources. The summer was exceptionally hot, with many days in July and August when temperatures exceeded 30 °C. The heat combined with low rainfall led to extreme drought. Meanwhile, July and August are the months of mating activity for roe deer. The late autumn rut allowed the roe deer-a monoestrous species-to limit the consequences of a decrease in mating activity or fertility during the hot summer. Global warming may affect roe deer reproduction, so climatic conditions should be considered in population studies, not only in terms of food availability.

Insights into archaeological and modern neotropical biomes: Examining diet and shape variation through white-tailed deer lower third molar

The white-tailed deer (Odocoileus virginianus Zimmermann 1780) holds significant ecological importance across the Americas, both historically and in modern times. This species ranges from southern Canada to Brazil and exhibits polytypic characteristics, adapting well to diverse habitats including temperate, subtropical, semi-arid, rainforest, and savanna environments. In paleontology and archaeology, the comparison of dental characteristics between extant mammal species with known diets is commonly employed to infer the feeding behaviors of their ancient counterparts. This method assumes that extant and fossil species share similar dietary preferences, aiding in the identification of past environmental contexts. Consequently, we employed a multiproxy approach, combining the study of dental wear and 2D geometric morphometrics, to investigate potential relationships between molar shape, diet, and biomes among extant white-tailed deer populations across the Americas. Our analysis included a comparison with archaeological data from Panama. We sampled 274 extant lower second molar specimens for micro- and mesowear analysis, along with 105 lower third molar specimens from natural science museums for 2D geometric morphometric analysis. These were compared with a sample of 65 archaeological specimens from Panama. Our findings revealed distinct variations in the shape of lower m3 molars among extant white-tailed deer populations across different biomes, with notable differences observed in the archaeological samples as well. Micro- and mesowear analyses also indicated biome-related differences, suggesting a general browsing diet for white-tailed deer with nuanced variations across biomes. Mesowear analysis further suggested a dietary spectrum ranging from pure browsers to browser-mixed feeders. These findings offer valuable insights for the interpretation of fossil deer specimens recovered from archaeological sites.

Demographic Risk Factors Vary in the Invasion Front of Chronic Wasting Disease in West Virginia, USA.

After detecting chronic wasting disease (CWD) in white-tailed deer (Odocoileus virginianus) in Hampshire County, West Virginia, USA, in 2005, we investigated the change of CWD apparent prevalence and potential factors influencing infection risk during the invasion front. Over eight sampling years (2006-2012 and 2017) during a 12-yr period within a 101-km2-area monitoring zone, we sampled and tested a total of 853 deer for CWD by ELISA and immunohistochemistry. Bayesian logistic regression of risk factors included collection year, age class, sex, and adjusted body weight (weight after accounting for sex, age, kidney fat index, and number of fetuses). In the whole-herd model (n=634), collection year, age, and adjusted body weight were associated with increased odds of CWD, whereas an age-weight interaction had a negative relationship. We found that males drove the positive associations with age and adjusted body weight, whereas females were responsible for the negative interaction effect. These findings suggest potential behavioral and physiological mechanisms related to sex that may influence CWD exposure. Older males exhibited higher CWD prevalence, aligning with previous studies. Notably, the novel finding of adjusted body weight as a risk factor in males warrants further investigation, and this study highlights the need for future research on social behavior and its role in CWD transmission within white-tailed deer populations.

Montane willow scrub restoration in Scotland: reviewing 30 years of progress to reestablish the altitudinal treeline

Many mountain woodlands across the globe have undergone anthropogenic degradation with negative implications for the ecological and socioeconomic systems dependent on them. Montane willows are arctic‐alpine species of high nature conservation concern within the altitudinal treeline. Scotland has been pioneering the restoration of montane willow scrub for three decades following widespread loss linked particularly to overgrazing by sheep and deer since the eighteenth century. In the 1990s, the total area of montane willows throughout the country was less than 10 ha, with most remaining populations highly fragmented, in rapid decline, and restricted to inaccessible cliffs. Subsequently, montane willow scrub restoration has been part of an overall 2659 ha area managed for the recovery of mountain woodlands in Scotland. By May 2023, 396,868 montane willows were planted in projects encompassing large‐scale habitat creation, relict population reinforcement by genetic rescue, fencing, and landscape‐scale management for low‐density deer populations. This review summarizes restoration methods and outcomes with case study examples to promote conservation evidence and research on plant growth and survival, planting sites, associated biodiversity, molecular ecology, genomics, and long‐term population sustainability. With the potential for montane scrub restoration to be widely expanded, improved connectivity to revive a treeline mosaic shaped by regeneration will tackle biodiversity loss and help mitigate climate change impacts on a national scale. However, caution must be applied to creating definitions of distinct habitats for restoration that are based on non‐natural or ecologically degraded systems. These experiences are particularly relevant to montane and circumpolar regions with strongly modified landscape histories.

Body mass change over winter is consistently sex-specific across roe deer (Capreolus capreolus) populations

Body mass change over winter is consistently sex-specific across roe deer (Capreolus capreolus) populations

Annual variation in recruitment rates for the sika deer (Cervus nippon) population in eastern Hokkaido, Japan

Annual variation in recruitment rates for the sika deer (<i>Cervus nippon</i>) population in eastern Hokkaido, Japan

One Health collaboration is more effective than single-sector actions at mitigating SARS-CoV-2 in deer

One Health aims to achieve optimal health outcomes for people, animals, plants, and shared environments. We describe a multisector effort to understand and mitigate SARS-CoV-2 transmission risk to humans via the spread among and between captive and wild white-tailed deer. We first framed a One Health problem with three governance sectors that manage captive deer, wild deer populations, and public health. The problem framing included identifying fundamental objectives, causal chains for transmission, and management actions. We then developed a dynamic model that linked deer herds and simulated SARS-CoV-2. Next, we evaluated management alternatives for their ability to reduce SARS-CoV-2 spread in white-tailed deer. We found that single-sector alternatives reduced transmission, but that the best-performing alternative required collaborative actions among wildlife management, agricultural management, and public health agencies. Here, we show quantitative support that One Health actions outperform single-sector responses, but may depend on coordination to track changes in this evolving system.

Using Unmanned Aerial Vehicles Equipped with Thermal Cameras to Survey a Known Population of White-Tailed Deer

Abstract New technologies are emerging that may have potential for yielding more reliable population estimates for white-tailed deer Odocoileus virginianus than traditional manned aerial surveys. In a study conducted in a 116.55-ha enclosure in the Edwards Plateau ecoregion of Texas, USA, we tested the accuracy and precision of population counts by using an unmanned aerial vehicle equipped with a dual thermal–red green blue (heat detection and color video) camera to conduct a complete area count of a known, closed population of white-tailed deer. We flew surveys during 2–4 December 2019 in preprogrammed transects and replicated them three times, in both the morning and afternoon. Detection was higher during morning surveys, when we detected almost all deer that were present, compared with afternoon surveys. The accuracy and precision of morning surveys reported herein justify further research into the development of a method using this equipment for surveying large mammal populations in certain habitats.

Comparison of thermal cameras and human observers to estimate population density of fallow deer (Dama dama) from aerial surveys in Tasmania, Australia

Context The population of introduced fallow deer (Dama dama) is thought to have increased exponentially across much of the island of Tasmania, Australia, since 2000. Historically, deer management decisions have relied on population trend data from vehicular spotlight surveys. Renewed focus on the contemporary management of the species requires development of more robust and precise population estimation methodology. Aims This study demonstrates two aerial survey methods – conventional counts by trained human observers, and thermal imaging footage recorded during the same flights – to inform future survey practices. Methods Conventional counts were carried out by three observers, two seated on the left side of the helicopter, and one on the right. A high-resolution thermal camera was fitted to the helicopter and was orientated to meet the assumptions of distance sampling methodologies. Both survey methods were used to generate deer population density estimates. Spatial distribution of deer was also analysed in relation to patches of remnant native vegetation across an agricultural landscape. Mark–recapture distance sampling was used to estimate density from human observer counts and provide a comparison to the distance sampling estimates derived from the thermal camera. Key results Human observer counts gave a density estimate of 2.7 deer per km2, while thermal camera counts provided an estimate of 2.8 deer per km2. Deer population density estimates calculated via both methods were similar, but variability of the thermal camera estimate (coefficient of variation (CV) of 36%) was unacceptably high. Human observer data was within acceptable bounds of variability (CV, 19%). The estimated population size in central and north-eastern Tasmania for 2019 approximated 53,000 deer. Deer were primarily congregated within 200 metres of the interface between canopy cover and open pasture. Conclusions The population density estimate provides a baseline for monitoring and managing the Tasmanian deer population. Human observer data was more precise than thermal camera data in this study, but thermal counts could be improved by reducing sources of variability. Implications Improvements for the collection of thermal imagery are recommended. Future control efforts may be more efficient if they preferentially target habitat edges at this time of year, paired with random or grid-based searches where population density is lower.

Positive impact of large wild herbivore exclusion on silver fir regeneration: A case study from the Poľana Mountains, Central Slovakia

Abstract In forest ecosystems, the browsing activity of large wild herbivores (LWH) often leads to reduced tree and plant diversity, diminished biomass production, and challenges in achieving forest management objectives. Our case study focuses on assessing the impact of LWH browsing during the initial stages of forest growth by comparing fenced plots (F plots) with excluded LWH and control plots (C plots) with the presence of LWH. The experiment took place at the Hukavský Grúň research site in the Poľana Mts., characterized by a high red deer (Cervus elaphus L.) population. Fifteen F plots and fifteen C plots, all situated within mixed maternal forests, were established in the 2023 growing season. Thus, circular plots with a radius of 2.5 m were utilized, and comprehensive data were collected on young trees (excluding those under 10 cm in height), covering tree species, positions, heights, and stem diameters. Species-specific allometric relations were employed to calculate the aboveground biomass of each tree, contributing to the overall biomass stock on a plot basis. A comparison between F and C plots revealed a prevalence of silver fir (Abies alba Mill.) in F plots, while European beech (Fagus sylvatica L.) dominated C plots. F plots exhibited higher tree species diversity (4.5 species), contrasting with the lower diversity (2.0 species) and absence of silver fir in C plots. The F plots also demonstrated greater tree density and sizes, resulting in substantial differences in aboveground biomass stocks. Browsing in C plots predominantly affected tree height rather than stem diameter, leading to a bigger height-to-diameter ratio in F plots compared to C plots. We suggest that fencing as a method to exclude LWH might be economically expensive and provide a temporary solution limited by the functionality of the fence. Therefore, the primary strategy for safeguarding the future of silver fir may lie in regulating LWH populations to a reasonable threshold.