Plasma and interstitial fluid antibiotic levels of subcutaneously implanted compounded florfenicol calcium sulfate beads in New Zealand White rabbits (Oryctolagus cuniculus).

OBJECTIVE: To determine antibiotic levels in plasma and interstitial fluid (ISF) after SC placement of compounded florfenicol (FF) calcium sulfate beads (CSBs) in New Zealand White rabbits (Oryctolagus cuniculus). ANIMALS: 6 juvenile female rabbits (n = 5 treatment and 1 control). METHODS: An ultrafiltration probe and CSBs were placed SC in 6 rabbits (n = 5 for FF CSBs and 1 for control CSBs). Plasma (3, 6, 12, 24, and 48 hours and 7, 14, and 21 days) and ISF (daily for 21 days) samples were collected, and FF was measured by HPLC for pharmacokinetic analysis. Hematology, biochemistry, and histopathology were assessed. RESULTS: Means ± SD for the area under the curve, maximum concentration, time of maximum concentration, terminal half-life, and mean residence time to the last data point for plasma and ISF were 16.63 ± 8.16 and 17,902 ± 7,564 h·µg/mL, 0.79 ± 0.38 and 245 ± 223 µg/mL, 2.90 ± 0.3 and 59 ± 40 hours, 30.81 ± 16.9 and 27.3 ± 9.39 hours, 23.4 ± 10 and 73.7 ± 13 hours, respectively. Plasma FF was < 2 µg/mL at all time points. The ISF FF remained > 8 µg/mL for 109.98 to 231.58 hours. One rabbit death occurred during treatment, but the cause of death was undetermined. Local tissue inflammation was present, but no clinically significant systemic adverse effects were found on hematology, biochemistry, or histopathology in the remaining rabbits. CLINICAL RELEVANCE: Florfenicol CSBs maintained antibiotic concentrations in ISF at levels likely to be effective against bacteria sensitive to > 8 µg/mL for 5 to 10 days while maintaining low (< 2 µg/mL) plasma levels. Florfenicol CSBs may be effective for local antibiotic treatment in rabbit abscesses.

First Report of Chagas Disease Vector Species Triatoma sanguisuga (Hemiptera: Reduviidae) Infected with Trypanosoma cruzi in Delaware.

In July and October 2023, two live triatomine bugs were found inside a home in New Castle County, Delaware. The bugs were identified as Triatoma sanguisuga, the most widespread triatomine bug species in the United States. Triatoma sanguisuga is a competent vector of Trypanosoma cruzi, the causative agent of Chagas disease. The two specimens were tested via real-time PCR (qPCR) for infection with T. cruzi, and one of the specimens was positive. Despite T. sanguisuga being endemic to the area, attainment of accurate species identification and T. cruzi testing of the bugs required multiple calls to federal, state, private, and academic institutions over several months. This constitutes the first report of T. sanguisuga infected with T. cruzi in Delaware. In addition, this is the first published report of T. sanguisuga in New Castle County, the northernmost and most densely populated county in Delaware. New Castle County still conforms to the described geographic range of T. sanguisuga, which spans from Texas to the East Coast of the United States. The T. cruzi infection prevalence of the species has not been studied in the northeastern United States, but collections in southern states have found prevalences as high as 60%. The Delaware homeowner's lengthy pursuit of accurate information about the vector highlights the need for more research on this important disease vector in Delaware.

Detection of skunk adenovirus-1 in domestic ferrets (Mustela putorius).

We describe gross pathology, histopathology, ultrastructure, and molecular characterization of skunk adenovirus 1 (SkAdV-1) in 3 ferrets (Mustela putorius) submitted to the Infectious Diseases Laboratory, Zoo and Exotic Animal Pathology Service at the University of Georgia. Case 1 came from a cohort with a 6-week history of multiple ferrets with respiratory disease and mortality. Cases 2 and 3 were submitted 19 days later; they died 3 days after arrival at the vendor's facility. Histopathological evaluation found evidence of suppurative bronchopneumonia, with intralesional gram-positive cocci in case 1. The others had evidence of subacute multifocal histiocytic and lymphoplasmacytic interstitial pneumonia. Viral isolation, polymerase chain reaction (PCR), and DNA sequencing identified SkAdV-1 in lung tissue. In situ hybridization confirmed adenovirus within the lung lesion, and adenovirus particles were visible under an electron microscope. This is the seventh species in which SkAdV-1 has been identified. Cross-species transmission events have important implications for wildlife management and multispecies facilities.

Angiostrongylus cantonensis Infection in Brown Rats (Rattus norvegicus), Atlanta, Georgia, USA, 2019-2022.

Rat lungworm (Angiostrongylus cantonensis), a zoonotic parasite invasive to the United States, causes eosinophilic meningoencephalitis. A. cantonensis harbors in rat reservoir hosts and is transmitted through gastropods and other paratenic hosts. We discuss the public health relevance of autochthonous A. cantonensis cases in brown rats (Rattus norvegicus) in Atlanta, Georgia, USA.

An Acute Mortality Event Associated with Novel Macrorhabdus ornithogaster Infection and Underlying Factors in a Newly-Established Captive Group of American White Ibis (Eudocimus albus) Nestlings.

Twenty-four American white ibis (Eudocimus albus) nestlings were collected in Florida (USA) on 17 April 2017 to establish a captive flock. On 7 May 2017, three birds died suddenly, following severe lethargy, hemorrhaging from the mouth and nares, anorexia, and production of bright-green colored feces. An additional ibis with delayed growth and pathological fractures was euthanized 18 May 2017. Severe ventriculitis associated with Macrorhabdus ornithogaster was noted in all four birds, bacterial sepsis was confirmed in one bird by culture and histologic examination, and bacterial endotoxemia was suspected in two birds based on gross and histologic examination, but no bacteria were isolated from these birds. Birds also had vitamin E liver levels consistent with coagulopathy previously described in pelicans. We sampled feces from 91 adult, free-living, healthy ibis in Florida in July 2017 and found 71% were shedding organisms with morphologic characteristics consistent with Macrorhabdus sp. Molecular characterization of the ibis-origin M. ornithogaster showed it was phylogenetically related to numerous M. ornithogaster sequences. It is unknown if M. ornithogaster infection resulted in clinical disease as a result of dietary or stress-related dysbiosis, or other factors. Macrorhabdus-associated disease has not previously been confirmed in wading birds. We discuss potential associations of gastric M. ornithogaster infection with morbidity and mortality in these cases and highlight the need for additional studies on this pathogen in free-living birds.

Drivers of African Filovirus (Ebola and Marburg) Outbreaks.

Outbreaks of African filoviruses often have high mortality, including more than 11,000 deaths among 28,562 cases during the West Africa Ebola outbreak of 2014-2016. Numerous studies have investigated the factors that contributed to individual filovirus outbreaks, but there has been little quantitative synthesis of this work. In addition, the ways in which the typical causes of filovirus outbreaks differ from other zoonoses remain poorly described. In this study, we quantify factors associated with 45 outbreaks of African filoviruses (ebolaviruses and Marburg virus) using a rubric of 48 candidate causal drivers. For filovirus outbreaks, we reviewed >700 peer-reviewed and gray literature sources and developed a list of the factors reported to contribute to each outbreak (i.e., a "driver profile" for each outbreak). We compare and contrast the profiles of filovirus outbreaks to 200 background outbreaks, randomly selected from a global database of 4463 outbreaks of bacterial and viral zoonotic diseases. We also test whether the quantitative patterns that we observed were robust to the influences of six covariates, country-level factors such as gross domestic product, population density, and latitude that have been shown to bias global outbreak data. We find that, regardless of whether covariates are included or excluded from models, the driver profile of filovirus outbreaks differs from that of background outbreaks. Socioeconomic factors such as trade and travel, wild game consumption, failures of medical procedures, and deficiencies in human health infrastructure were more frequently reported in filovirus outbreaks than in the comparison group. Based on our results, we also present a review of drivers reported in at least 10% of filovirus outbreaks, with examples of each provided.

Cat Scratch Disease: 9 Years of Experience at a Pediatric Center.

Background: A more complete understanding of the epidemiology, risk factors, and clinical features of cat scratch disease (CSD) in children could help guide patient care. Methods: We conducted a retrospective analysis of children presenting to a tertiary pediatric hospital system in Atlanta, Georgia between January 1, 2010 and December 31, 2018 who had serology, polymerase chain reaction, and/or cytopathological results consistent with a Bartonella henselae infection. We also retrospectively reviewed veterinary diagnostic results performed at the University of Georgia from 2018 to 2020 to ascertain the burden of bartonellosis in companion animals within the state. Results: We identified 304 children with CSD over 9 years with the largest proportion of diagnoses made during August (41 of 304, 13.5%) and September (47 of 304, 15.5%). The median age of child cases was 8.1 years (interquartile range [IQR], 5.4-12.1); 156 (51.3%) were female; 242 of 262 (92.4%) reported feline exposure; and 55 of 250 (22%) reported canine exposure of those with exposure histories documented in the medical record. Although lymphadenopathy was present on physical examination in the majority of cases (78.8%), atypical presentations lacking lymphadenopathy were also common (63 of 304, 20.7%). Among children with radiographic imaging, 20 of 55 (36.4%) had splenomegaly and 21 of 55 (38.1%) had splenic and/or hepatic microabscesses. Among veterinary data, Bartonella seroprevalence was 12 of 146 (8.2%), all among canines, with a geographic distribution that spanned the state of Georgia. Conclusions: Distinguishing clinical features of CSD included subacute regional lymphadenopathy in school-aged children in the late summer, almost all of whom had cat exposure. Atypical clinical manifestations of CSD were also commonly identified.

Land reversion and zoonotic spillover risk.

Deforestation alters wildlife communities and modifies human-wildlife interactions, often increasing zoonotic spillover potential. When deforested land reverts to forest, species composition differences between primary and regenerating (secondary) forest could alter spillover risk trajectory. We develop a mathematical model of land-use change, where habitats differ in their relative spillover risk, to understand how land reversion influences spillover risk. We apply this framework to scenarios where spillover risk is higher in deforested land than mature forest, reflecting higher relative abundance of highly competent species and/or increased human-wildlife encounters, and where regenerating forest has either very low or high spillover risk. We find the forest regeneration rate, the spillover risk of regenerating forest relative to deforested land, and how rapidly regenerating forest regains attributes of mature forest determine landscape-level spillover risk. When regenerating forest has a much lower spillover risk than deforested land, reversion lowers cumulative spillover risk, but instaneous spillover risk peaks earlier. However, when spillover risk is high in regenerating and cleared habitats, landscape-level spillover risk remains high, especially when cleared land is rapidly abandoned then slowly regenerates to mature forest. These results suggest that proactive wildlife management and awareness of human exposure risk in regenerating forests could be important tools for spillover mitigation.

Pathology in Practice.

In collaboration with the American College of Veterinary Pathologists.

Both consumptive and non-consumptive effects of predators impact mosquito populations and have implications for disease transmission.

Predator-prey interactions influence prey traits through both consumptive and non-consumptive effects, and variation in these traits can shape vector-borne disease dynamics. Meta-analysis methods were employed to generate predation effect sizes by different categories of predators and mosquito prey. This analysis showed that multiple families of aquatic predators are effective in consumptively reducing mosquito survival, and that the survival of Aedes, Anopheles, and Culex mosquitoes is negatively impacted by consumptive effects of predators. Mosquito larval size was found to play a more important role in explaining the heterogeneity of consumptive effects from predators than mosquito genus. Mosquito survival and body size were reduced by non-consumptive effects of predators, but development time was not significantly impacted. In addition, Culex vectors demonstrated predator avoidance behavior during oviposition. The results of this meta-analysis suggest that predators limit disease transmission by reducing both vector survival and vector size, and that associations between drought and human West Nile virus cases could be driven by the vector behavior of predator avoidance during oviposition. These findings are likely to be useful to infectious disease modelers who rely on vector traits as predictors of transmission.

Mosquitoes are often referred to as the deadliest animals on earth because some species spread malaria, West Nile virus or other dangerous diseases when they bite humans and other animals. Adult mosquitoes fly to streams, ponds and other freshwater environments to lay their eggs. When the eggs hatch, the young mosquitoes live in the water until they are ready to grow wings and transform into adults. In the water, the young mosquitoes are particularly vulnerable to being eaten by dragonfly larvae, fish and other predators. When adult females are choosing where to lay their eggs, they can use their sense of smell to detect these predators and attempt to avoid them. Along with eating the mosquitoes, the predators may also reduce mosquito populations in other ways. For example, predators can disrupt feeding among young mosquitoes, which may affect the time that it takes for them to grow into adults or the size of their bodies once they reach the adult stage. Although the impacts of different predators have been tested separately in multiple settings, the overall effects of predators on the ability of mosquitoes to spread diseases to humans remain unclear. To address this question, Russell, Herzog et al. used an approach called meta-analysis on data from previous studies. The analysis found that along with increasing the death rates of mosquitoes, the presence of predators also leads to a reduction in the body size of those mosquitoes that survive, causing them to have shorter lifespans and fewer offspring. Russell, Herzog et al. found that one type of mosquito known as Culex ­ which carries West Nile virus ­ avoided laying its eggs near predators. During droughts, increased predation in streams, ponds and other aquatic environments may lead adult female Culex mosquitoes to lay their eggs closer to residential areas with fewer predators. Russell, Herzog et al. propose that this may be one reason why outbreaks of West Nile virus in humans are more likely to occur during droughts. In the future, these findings may help researchers to predict outbreaks of West Nile virus, malaria and other diseases carried by mosquitoes more accurately. Furthermore, the work of Russell, Herzog et al. provides examples of mosquito predators that could be used as biocontrol agents to decrease numbers of mosquitoes in certain regions.

Surveillance and genotype characterization of zoonotic trypanosomatidae in Didelphis marsupialis in two endemic sites of rural Panama.

Didelphis marsupialis has been reported as a competent reservoir for trypanosomatid parasites infections. The aim of this study was to measure Trypanosoma cruzi, T. rangeli, and Leishmania spp. infection rates and to characterize discrete typing units (DTUs) of T. cruzi in D. marsupialis from two Chagas disease endemic sites in Panama. Blood from 57 wild-caught D. marsupialis were examined from two rural communities, Las Pavas (N = 18) and Trinidad de las Minas (N = 39). Twenty-two (38.60%) opossums were positive for flagellates by general hemoculture. T. cruzi infection was confirmed by positive hemoculture and/or kDNA based PCR performed in 31/57 (54.39%) blood samples from opossums. T. rangeli infection was confirmed by hemoculture and/or TrF/R2-Primer PCR assay applied on 12/57 (21.05%) blood samples. Nine (15.79%) D. marsupialis harbored T. cruzi/T. rangeli coinfections. All opossums tested negative for Leishmania spp. by PCR assays based on kDNA and HSP70 gene amplification. There was a significant association between T. cruzi infection and site (Fisher exact test, p = 0.02), with a higher proportion of T. cruzi infected opossums in Las Pavas (77.78%, n = 14/18) compared to Trinidad de las Minas (43.59%, n = 17/39). A significant association was found between habitat type and T. cruzi infection in opossums across both communities, (X2 = 6.91, p = 0.01, df = 1), with a higher proportion of T. cruzi infection in opossums captured in forest remnants (76%, 19/25) compared to peridomestic areas (37.5%, 12/32). T. rangeli detection, but not T. cruzi detection, may be improved by culture followed by PCR. TcI was the only DTU detected in 22 T. cruzi samples using conventional and real-time PCR. Eight T. rangeli positive samples were characterized as KP1(-)/lineage C. Trypanosome infection data from this common synanthropic mammal provides important information for improved surveillance and management of Chagas disease in endemic regions of Panama.

TEMPORAL AND SPATIAL PATTERNS IN CANINE DISTEMPER VIRUS CASES IN WILDLIFE DIAGNOSED AT THE SOUTHEASTERN COOPERATIVE WILDLIFE DISEASE STUDY, 1975-2019.

Canine distemper is a high-impact disease of many mammal species and has caused substantial carnivore population declines. Analysis was conducted on passive surveillance data of canine distemper (CDV)-positive wild mammal cases submitted to the Southeastern Cooperative Wildlife Disease Study, Athens, Georgia, US, between January 1975 and December 2019. Overall, 964 cases from 17 states were CDV positive, including 646 raccoons (Procyon lotor), 254 gray foxes (Urocyon cinereoargenteus), 33 striped skunks (Mephitis mephitis), 18 coyotes (Canis latrans), four red foxes (Vulpes vulpes), three gray wolves (Canis lupus), three American black bears (Ursus americanus), two American mink (Mustela vison), and one long-tailed weasel (Mustela frenata). Raccoon and gray fox case data from the state of Georgia (n=441) were selected for further analysis. Autoregressive integrated moving average models were developed predicting raccoon and gray fox case numbers. The best-performing model for gray foxes used numbers of gray fox CDV cases from the previous 2 mo and of raccoon cases in the present month to predict the numbers of gray fox cases in the present month. The best-performing model for raccoon prediction used numbers of raccoon CDV cases from the previous month and of gray fox cases in the present month and previous 2 mo to predict numbers of raccoon cases in the present month. Temporal trends existed in CDV cases for both species, with cases more likely to occur during the breeding season. Spatial clustering of cases was more likely to occur in areas of medium to high human population density; fewer cases occurred in both the most densely populated and sparsely populated areas. This pattern was most prominent for raccoons, which may correspond to high transmission rates in suburban areas, where raccoon population densities are probably highest, possibly because of a combination of suitable habitat and supplemental resources.

Reproductive tract neoplasia in adult female Asian elephants (Elephas maximus).

Recent reports have highlighted a lower-than-expected prevalence of neoplasia in elephants and suggested mechanisms for cancer resistance. But despite infrequent reports in the literature, uterine neoplasia is common in managed Asian elephants (Elephas maximus). This study is an archival review of reproductive tract neoplasia in 80 adult female Asian elephant mortalities in managed care facilities in the United States from 1988 to 2019. Neoplasms occurred in 64/80 (80%) of cases. Most were in the uterus (63/64; 98%) with only a single case of ovarian neoplasia. Myometrial leiomyomas were present in 57/63 (90%) cases with uterine neoplasia. Uterine adenocarcinoma was present in 8/63 (13%) cases. Remaining cases included endometrial adenoma (2), focal carcinoma in situ in endometrial polyps (1), anaplastic carcinoma (1), endometrial hemangioma (1), primitive neuroectodermal tumor (PNET; 1), and angiosarcoma (1). One case with uterine adenocarcinoma had a separate pelvic mass histologically characterized as an anaplastic sarcoma. Distant metastases were documented in 5/8 (63%) cases of uterine adenocarcinoma, and in the uterine anaplastic carcinoma, PNET, and angiosarcoma. Four uterine adenocarcinomas and one carcinoma in situ were examined immunohistochemically for pan-cytokeratin, vimentin, and estrogen receptor. In all, neoplastic cells were pan-cytokeratin positive and vimentin negative, and in 2 cases were immunoreactive for estrogen receptor. Results show that female reproductive tract neoplasia, particularly of the uterus, is common in Asian elephants and is not limited to leiomyomas. Importantly, uterine neoplasms have the potential to impact fecundity and may represent obstacles to conservation in managed care.

A Retrospective Study of Pathology in Bats Submitted to an Exotic and Zoo Animal Diagnostic Service in Georgia, USA (2008-2019).

Pathology records of bats submitted to the University of Georgia from managed care settings were reviewed to identify naturally occurring diseases. Fifty-nine cases were evaluated during an 11-year period (2008-2019), including representatives from four families: Pteropodidae (Yinpterochiroptera), Phyllostomidae, Vespertilionidae and Molossidae (Yangochiroptera). Pathology reports were reviewed to determine the primary pathological process resulting in death or the decision to euthanize. Cases were categorized as non-infectious (34/59; 58%), infectious/inflammatory (17/59; 29%) or undetermined due to advanced autolysis (8/59; 14%). Musculoskeletal diseases and reproductive losses were the most frequent pathological processes. Among the infectious processes identified, bacterial infections of the reproductive and haemolymphatic systems were most frequently observed. The first two reports of neoplasia in small flying foxes (Pteropus hypomelanus) are described. Bats under managed care present with a wide range of histopathological lesions. In this cohort, non-infectious disease processes were common.

Landscape disturbance impacts on Attalea butyracea palm distribution in central Panama.

BACKGROUND: Increased Attalea butyracea palm propagation, notable for its role as key habitat for the primary Chagas disease vector in Panama, has been linked to landscape disturbance in single-palm observations in this region. Close proximity of these palms to human dwellings is proposed to increase risk of Chagas disease transmission from sylvatic transmission cycles to domestic transmission involving human populations. This study examines the relationship between landscape disturbance and mature A. butyracea spatial distribution, density, and proximity to human populations and vector and reservoir species' movement corridors at a regional scale in a 300 km2 heterogeneous tropical landscape in central Panama. METHODS: We remotely identified the locations of over 50,000 mature A. butyracea palms using high-resolution WorldView2 satellite imagery. A local Getis-Ord Gi* spatial analysis identified significant clusters of aggregated palms. Associations between palm and cluster abundance and a landscape disturbance gradient, derived from official Panama land cover data, were tested using Chi-square tests for Homogeneity and Z-test for proportions. Kruskall-Wallis non-parametric analysis of variance tests were run to assess whether palm cluster area varied by disturbance level, or whether disturbance was associated with proximity of palms and palm clusters to susceptible populations or vector movement corridors. RESULTS: Our findings indicate a regional relationship between landscape disturbance and A. butyracea occurrence. We observe a significant increase in both individual and clustered A. butyracea in secondary forest, but a reduction of palms in agricultural settings. We do not detect evidence of any reduction in abundance of palms in residential settings. The majority of residential and commercial buildings in our study area are within vector flight distance of potential vector habitat in palm crowns. CONCLUSIONS: We observe probable anthropogenic elimination of A. butyracea palms in agricultural, but not residential, settings. Even in heavily deforested regions, significant concentrations of mature palms remain in close proximity to human establishments.

Granulomatous Inflammation of the Muzzle in White-Tailed Deer (Odocoileus virginianus) and Mule Deer (Odocoileus hemionus) Associated With Mannheimia granulomatis.

Since 2002, reports of deer with swollen muzzles from throughout the United States have resulted in significant interest by wildlife biologists and wildlife enthusiasts. The condition was identified in 25 white-tailed deer (Odocoileus virginianus) and 2 mule deer (O. hemionus). Microscopic lesions consisted of severe, granulomatous or pyogranulomatous inflammation of the muzzle, nasal planum, and upper lip, as well as similar but less severe inflammation of the hard palate. Lymphadenitis of regional lymph nodes was common and granulomatous pneumonia was present in one individual. Splendore-Hoeppli material was typical in the center of inflammatory foci. Other than the single instance of pneumonia, systemic disease was not evident. Various bacterial species were isolated in culture, most of which were not morphologically consistent with the colonies of small, gram-negative bacteria observed in the center of the granulomas. Amplification and sequencing of the bacterial 16S rRNA gene from tissues of affected deer resulted in the identification of Mannheimia granulomatis. Laser capture microdissection was used to confirm that the colonies in the inflammatory foci were M. granulomatis. The cases described here are reminiscent of a bovine disease in Brazil and Argentina, locally called lechiguana. Although the inflammation of lechiguana is mostly truncal, the microscopic lesions are very similar and are also attributed to M. granulomatis. It is unclear if this is an emerging infectious disease of deer, or if it is a sporadic, uncommon condition that has only recently been recognized.

How microclimatic variables and blood meal sources influence Rhodnius prolixus abundance and Trypanosoma cruzi infection in Attalea butyracea and Elaeis guineensis palms?

Chagas disease is a zoonosis that affects several million people and is caused by the parasite Trypanosoma cruzi, which is mainly transmitted through the feces of triatomine bugs. Within triatomines, several Rhodnius species have been found inhabiting palms, and certain factors such as palm species and location have been related to the abundance and T. cruzi infection of those insects in palms. In this study, the main goal was to determine if R. prolixus abundances and infection rates in Attalea butyracea and Elaeis guineensis palms are related to ecological factors such as palm species, crown microclimate, and available blood meal sources. Triatomine sampling was performed in two municipalities of Casanare, Colombia, specifically in the intersection of riparian forests and oil palm plantations. For R. prolixus abundance per palm, the predictors showing more relationship were palm species and blood meal species identified in the palm, and for T. cruzi infection per triatomine, they were palm species and nymphal stage. Palm microclimate was very similar in both palm species and did not show a relationship with triatomine abundance. Comparing palm species, A. butyracea showed more blood meal species, including more refractory host species, than E. guineensis, but lower T. cruzi infection rate and parasitaemia. Interestingly, non-arboreal blood meal species were frequently found in the analyzed nymphs, indicating that the blood source for R. prolixus in palms corresponded to all the fauna located in the surrounded landscape and not only in the palm. These results could expose a new ecological scenario to interpret the T. cruzi transmission in sylvatic environments.