Biomonitoring Organochlorine Pesticides in Didelphis virginiana from Yucatan, Mexico by GC-ECD.

The aim of the present work was to apply a validated methodology for the detection of organochlorine pesticides in Didelphis virginiana (Virginia opossum) serum samples collected in Yucatan, Mexico. Recent studies performed to investigate the presence of Organochlorines (OCLs) in water, human blood and milk, and animal tissues from Yucatan have shown that the OCLs concentrations are high and can be associated with potential human health risk. Since opossum is considered an important synanthropic species in Yucatan, 40 opossum serum samples were analyzed by gas chromatography with electron capture detector. The most common OCLs found in opossum sera were lindanes, chlordanes, drines, and endosulfan. Heptachlor, heptachlor epoxide, and lindanes were found at the highest concentrations, while dichlorodiphenyl trichloroethane and its metabolites were found at the lowest concentrations in the samples. The good linearity, precision, and accuracy obtained in the evaluated parameters in the extraction and chromatographic methods support its application for the monitoring of OCLs pesticides in populations of opossums and other wild species in Yucatan.

PHYSIOLOGICAL BASAL PARAMETERS OF FREE-RANGING OPOSSUMS ( DIDELPHIS VIRGINIANA) IN THE YUCATAN PENINSULA, MEXICO.

The aim of the present study is to determine the basal parameters in opossums ( Didelphis virginiana) during physical examination in Yucatan, Mexico. Sixty-six opossums were captured and manually handled for physical examinations. Sex, age, cardiac and respiratory rate, body temperature, pulse, mucous membranes, capillary refill time, hydration, size of superficial lymph nodes, mental status, and body condition were evaluated. Also, comparisons between rectal and auricular temperatures were performed. The means of physiological parameters obtained were: cardiac frequency 146 beats per minute (95% confidence interval [CI]: 138.91-155.24), respiratory frequency 27.33 breaths per minute (95% CI: 25.15-29.64), and body temperature 34.01°C (95% CI: 33.71-34.31). In 54% of animals, a sinus arrhythmia was present. A significant difference was found between rectal and auricular temperature measurements. A variation of -1.64 to 1.14°C was obtained via the auricular thermometer compared with the rectal device.

Prevalence, Abundance and Intensity of Eggs and Oocysts of Gastrointestinal Parasites in the Opossum Didelphis Virginiana Kerr, 1792 in Yucatan, Mexico.

Virginia opossum, Didelphis virginiana, is a synanthropic mammal associated with peridomestic areas of Yucatán, However, little is known about the gastrointestinal parasite infections of this species. The infection prevalence, mean abundance and mean intensity of eggs and oocysts of gastrointestinal parasites, in opossums captured in the peridomestic areas were estimated in six rural localities of Yucatán, Mexico. Eighty-four faecal samples were processed by flotation technique. McMaster test was used to estimate the number of helminth eggs and protozoa oocysts per gram of feces. Seven genera of gastrointestinal parasites were identified, and then infection prevalence was estimated as follows: Protozoa Eimeria sp. (51.9 %) and Sarcocystis sp. (1 %); nematodes Ancylostoma sp. (80.56 %), Cruzia sp. (62.04 %), Trichuris sp. (60.19 %), Capillaria sp. (29.63 %), Turgida sp. (23.15 %), Toxocara sp. (11.11 %), and Ascaris sp. (1.85 %); and one acanthocephalan: Oligacanthorhynchus sp. (14.81 %). This is the first study on the diversity of gastrointestinal parasites in Virginia opossums, and first evidence about the potential role of opossums in the transmission of zoonotic gastrointestinal parasites in peridomestic areas of Yucatán, Mexico.

PULMONARY LESIONS CAUSED BY THE LUNGWORM (DIDELPHOSTRONGYLUS HAYESI) IN THE OPOSSUM (DIDELPHIS VIRGINIANA) IN COLIMA, MEXICO.

Didelphostrongylus hayesi is an important and prevalent pulmonary nematode in the opossum ( Didelphis virginiana ). An in-depth description of the pulmonary lesions caused by this nematode is lacking. The objective of this investigation was to make a detailed account of the gross, subgross, and microscopic changes that occur in the lungs of opossums naturally infected with D. hayesi. Forty-four opossums trapped in the state of Colima, Mexico, were euthanized by an overdose of barbiturates. Following a postmortem examination, the right lung was cut from the main bronchi and placed in a Petri dish containing a saline solution for the detection and identification of live parasites. The left lung was fixed and cut serially for subgross microscopic examination and sections of lung were cut and stained for histopathologic examination. The most remarkable gross change in parasitized lungs was a poorly collapsible pulmonary parenchyma and mild emphysema. The right lung tested positive for lungworms on gross examination in 20/44, and 11/44 (25%) of the left lungs showed tan nodules on the pleural surface. Microscopically, the bronchi of 20/44 animals harbored adult and larval stages of D. hayesi (left lung), the same 20 opossums from which nematodes were grossly evident at necropsy (right lung). Adults and larvae were present in bronchi, bronchioles, and alveoli mixed with desquamated cells and many eosinophils, and to a lesser extent neutrophils, alveolar macrophages, and giant cells. Bronchi and bronchioles exhibited goblet cell hyperplasia and metaplasia respectively, and infiltration of lymphoplasmacytic cells in the interstitium and lamina propria. The tan nodules consisted of focal alveolar endogenous lipidosis, which likely resulted from parasitic airway obstruction. The lungs of 3/20 parasitized opossums also showed alveolar bronchiolization (Lambertosis). The absence of Eucoleus aerophilus or bacterial pneumonia incriminates D. hayesi as the putative cause of pulmonary lesions in these opossums.

Metal accumulation in wild-caught opossum.

The Virginia opossum (Didelphis virginiana) is widespread in the USA, ranging south through Latin America. The ecology of opossums is such that they are in frequent contact with soils, suggesting that they may function as a valuable bioindicator for chemical contamination in terrestrial environments. Surprisingly, there have been virtually no toxicology studies on opossums. Here, we provide the first analysis of metal contaminants in opossum liver tissues. Liver samples were obtained from 471 opossums, collected from 2003 to 2006, at four sites in North Florida and South Georgia, USA, and concentrations of copper, lead, nickel, selenium, and zinc were measured. We found little evidence of age differences in the concentration of any of the metals. However, there were at least some significant differences between years, males and females, and between sites for each metal, although the pattern of these differences was not always consistent across metals. Concentrations of metals in liver tissue were positively correlated with one another, primarily of each metal (except Pb) with zinc. Reference levels of metal contaminants are not available for opossums, but concentrations of Cu, Ni, Pb, and Zn in our samples were for the most part significantly higher than those reported from liver tissues of nine-banded armadillos (Dasypus novemcinctus) collected at the same sites and in the same years. Data from other small mammals studied elsewhere further indicate that metal concentrations in opossums were high, but at this time, it is not possible to determine if these elevated levels generated toxicity. The substantial temporal and spatial variation we found in metal concentrations suggests that determination of baseline levels for opossums may not be straightforward. Nonetheless, this is the first study quantifying metal accumulation in the livers of Didelphis virginiana and, as such, provides an important starting point for future research.

Molecular characterization and development of Sarcocystis speeri sarcocysts in gamma interferon gene knockout mice.

The North American opossum (Didelphis virginiana) is the definitive host for at least three named species of Sarcocystis: Sarcocystis falcatula, Sarcocystis neurona and Sarcocystis speeri. The South American opossums (Didelphis albiventris, Didelphis marsupialis and Didelphis aurita) are definitive hosts for S. falcatula and S. lindsayi. The sporocysts of these Sarcocystis species are similar morphologically. They are also not easily distinguished genetically because of the difficulties of DNA extraction from sporocysts and availability of distinguishing genetic markers. Some of these species can be distinguished by bioassay; S. neurona and S. speeri are infective to gamma interferon gene knockout (KO) mice, but not to budgerigars (Melopsittacus undulatus); whereas S. falcatula and S. lindsayi are infective to budgerigars but not to KO mice. The natural intermediate host of S. speeri is unknown. In the present study, development of sarcocysts of S. speeri in the KO mice is described. Sarcocysts were first seen at 12 days post-inoculation (p.i.), and they became macroscopic (up to 4 mm long) by 25 days p.i. The structure of the sarcocyst wall did not change from the time bradyzoites had formed at 50-220 days p.i. Sarcocysts contained unique villar protrusions, 'type 38'. The polymerase chain reaction amplifications and sequences analysis of three nuclear loci (18S rRNA, 28S rRNA and ITS1) and two mitochondrial loci (cox1 and cytb) of S. speeri isolate from an Argentinean opossum (D. albiventris) confirmed its membership among species of Sarcocystis and indicated an especially close relationship to another parasite in this genus that employs opossums as its definitive host, S. neurona. These results should be useful in finding natural intermediate host of S. speeri.

CUTANEOUS EPITHELIOTROPIC T-CELL LYMPHOMA WITH METASTASES IN A VIRGINIA OPOSSUM (DIDELPHIS VIRGINIANA).

A 2-yr-old, captive, intact female Virginia opossum ( Didelphis virginiana ) with a 7-mo history of ulcerative dermatitis and weight loss was euthanatized for progressive worsening of clinical signs. Initially the opossum was treated with several courses of antibiotics, both topically and systemically; systemic nonsteroidal anti-inflammatory medication; and, later, systemic glucocorticoids, with no improvement in clinical signs. Histopathologic samples of skin lesions taken 3 mo into the course of disease revealed no evidence of neoplasia; however, cytologic samples of a skin lesion taken 5 mo into the course of disease revealed mature lymphocytes, and were suggestive of cutaneous lymphoma. Postmortem histopathology revealed neoplastic cells consistent with lymphoma; these were found in the haired skin of the forearm, axilla, hind limb, face, and lateral body wall, as well as the liver, kidney, axillary lymph node, heart, and spleen. Multifocal neutrophilic and eosinophilic ulcerative and necrotizing dermatitis and folliculitis of the haired skin were also present. To the authors' knowledge, this is the first documented case of cutaneous lymphoma in a Virginia opossum and the first documented case with visceral metastases in a marsupial.

Venipuncture and Blood Collection on Conscious Virginia Opossums (Didelphis virginiana).

We developed a venipuncture technique of the ventral caudal vein in conscious Virginia opossums (Didelphis virginiana) in cage traps, enabling blood sampling without anesthesia. Blood samples were successfully collected from all 28 opossums in this study by this technique. Draw volume of <0.1 mL occurred in only four opossums (14%).

Seasonal Variation in Mammalian Mesopredator Spatiotemporal Overlap on a Barrier Island Complex.

Due to lack of apex predators in human-dominated landscapes, mesopredator relationships are complex and spatiotemporal niche partitioning strategies can vary, especially when seasonal shifts in resource availability occur. Our objective was to understand spatiotemporal niche overlap across seasons among mesopredators inhabiting a barrier island complex. We placed 19 unbaited cameras throughout Fort De Soto County Park, Florida, USA between February 2021 and July 2023. Of six mesopredator species detected, three species had >75 detections during both the wet and dry seasons (coyote, Canis latrans; Virginia opossum, Didelphis virginiana; and raccoon, Procyon lotor). Using general linear mixed models, we determined that during the wet season coyote-raccoon and raccoon-opossum detections were positively associated with each other (p < 0.05). During the dry season, raccoon-opossum detections were positively associated, and opossums were more likely to be detected around mangroves. After calculating coefficients of overlap, we found all three species varied their temporal activity between seasons. During the dry season exclusively, all three mesopredators occupied different temporal niches. The park's isolated but developed nature has potentially led to a destabilized mesopredator community. Understanding seasonal mesopredator dynamics of Fort De Soto is particularly important because this park supports a high number of nesting shorebirds and sea turtles, which are known food sources for mesopredators.

Widespread exposure to anticoagulant rodenticides among common urban mesopredators in Chicago.

Anticoagulant rodenticides (ARs) are currently the most common method to control rats in cities, but these compounds also cause morbidity and mortality in non-target wildlife. Little attention has been focused on AR exposure among mesopredators despite their ecological role as scavengers and prey for larger carnivores, thus serving as an important bridge in the biomagnification of rodenticides in food webs. In this study, we sampled liver tissue from raccoons (Procyon lotor; n = 37), skunks (Mephitis mephitis; n = 15), and Virginia opossums (Didelphis virginiana; n = 45) euthanized by pest professionals and brown rats (Rattus norvegicus; n = 101) trapped in alleys in Chicago, USA to evaluate how often these species are exposed to ARs. We tested whether mesopredators had a higher prevalence of ARs and to more AR compounds compared to rats and calculated biomagnification factors (mean concentration in mesopredators/rats) as indicators of biomagnification. Of 93 sampled mesopredators, 100 % were exposed to at least one AR compound, mainly brodifacoum (≥80 %), and 79 % were exposed to multiple AR compounds. We also documented teal stomach contents consistent with the consumption of rat bait and altricial young tested positive to the same AR as their mother, suggesting mammary transfer. Of the 101 rats, 74 % tested positive to at least one AR compound and 32 % were exposed to multiple AR compounds. All mesopredator species had biomagnification factors exceeding 1.00 for brodifacoum (6.57-29.07) and bromadiolone (1.08-4.31). Our results suggest widespread exposure to ARs among urban mesopredators and biomagnification of ARs in mesopredators compared to rats. Policies that limit AR availability to non-target species, such as restricting the sale and use of ARs to licensed professionals in indoor settings, education on alternatives, and more emphasis on waste management may reduce health risks for urban wildlife and people in cities around the world.

Head posture impacts mammalian hyoid position and suprahyoid muscle length: implication for swallowing biomechanics.

Instantaneous head posture (IHP) can extensively alter resting hyoid position in humans, yet postural effects on resting hyoid position remain poorly documented among mammals in general. Clarifying this relationship is essential for evaluating interspecific variation in hyoid posture across evolution, and understanding its implications for hyolingual soft tissue function and swallowing motor control. Using Didelphis virginiana as a model, we conducted static manipulation experiments to show that head flexion shifts hyoid position rostrally relative to the cranium across different gapes. IHP-induced shifts in hyoid position along the anteroposterior axis are comparable to in vivo hyoid protraction distance during swallowing. IHP also has opposite effects on passive genio- and stylohyoid muscle lengths. High-speed biplanar videoradiography suggests Didelphis consistently swallows at neutral to flexed posture, with stereotyped hyoid kinematics across different head postures. IHP change can affect suprahyoid muscle force production by shifting their positions on the length-tension curve, and redirecting lines of action and the resultant force from supra- and infrahyoid muscles. We hypothesize that demands on muscle performance may constrain the range of swallowing head postures in mammals. This article is part of the theme issue 'Food processing and nutritional assimilation in animals'.

Molecular phylogeny supports invalidation of Didelphodiplostomum and Pharyngostomoides (Digenea: Diplostomidae) and reveals a Tylodelphys from mammals.

Alaria, Didelphodiplostomum and Pharyngostomoides are among genera of diplostomid digeneans known to parasitize mammalian definitive hosts. Despite numerous recent molecular phylogenetic studies of diplostomids, limited DNA sequence data is available from diplostomids parasitic in mammals. Herein, we provide the first 28S rDNA and cox1 mtDNA sequences from morphologically identified, adult specimens of Didelphodiplostomum and Pharyngostomoides. Newly generated 28S sequences were used to infer the phylogenetic interrelationships of these two genera among other major lineages of diplostomoideans. The phylogeny based on 28S and a review of morphology clearly suggests that Pharyngostomoides should be considered a junior synonym of Alaria, while Didelphodiplostomum should be considered a junior synonym of Tylodelphys. Pharyngostomoides procyonis (type species), Pharyngostomoides adenocephala and Pharyngostomoides dasyuri were transferred into Alaria as Alaria procyonis comb. nov., Alaria adenocephala comb. nov. and Alaria dasyuri comb. nov.; Didelphodiplostomum variabile (type species) and Didelphodiplostomum nunezae were transferred into Tylodelphys as Tylodelphys variabilis comb. nov. and Tylodelphys nunezae comb. nov. In addition, Alaria ovalis comb. nov. (formerly included in Pharyngostomoides) was restored and transferred into Alaria based on a morphological study of well-fixed, adult specimens and the comparison of cox1 DNA sequences among Alaria spp. The diplostomid genus Parallelorchis was restored based on review of morphology.

Urban foci of murine typhus involving cat fleas (Ctenocephalides felis felis) collected from opossums in Mexico City.

Murine typhus, a neglected rickettsiosis caused by Rickettsia typhi, is a common disease in several Latin-American countries. The sylvatic life cycle of R. typhi encompasses the presence of several wild mammals, particularly opossums of the genus Didelphis and their associated fleas. Due to the colonization of wild environments by human populations, the increase in contact with opossum fleas has generated the presence of urban outbreaks of typhus. For this reason, the aim of our study was to identify the presence and diversity of Rickettsia sp. in fleas collected from opossums of an urban reserve in Mexico City. Opossums were captured from February to September 2017. For the detection of Rickettsia DNA, fragments of 800 bp of the citrate synthase (gltA) and the outer membrane protein B (ompB) were amplified. A total of 141 fleas (111 ♀, 30 ♂) of a single species (Ctenocephalides felis felis) were recovered from 31 Didelphis virginiana. Rickettsia DNA was detected in 17.7% (25/141) of the analysed fleas, recovered from seven infested opossums. The Maximum likelihood of sequences exhibited an identity of 99%-100% with sequences of R. typhi from southern United States. This work represents the first record of R. typhi in fleas from opossums in Mexico.

Association of the invasive Haemaphysalis longicornis tick with vertebrate hosts, other native tick vectors, and tick-borne pathogens in New York City, USA.

Haemaphysalis longicornis, the Asian longhorned tick, is an invasive ixodid tick that has rapidly spread across the northeastern and southeastern regions of the United States since first reported in 2017. The emergence of H. longicornis presents a potential threat for livestock, wildlife, and human health as the host associations and vector potential of this invasive pest in the United States are poorly understood. Previous field data from the United States has shown that H. longicornis was not associated with natural populations of small mammals or birds, but they show a preference for medium sized mammals in laboratory experiments. Therefore, medium and large sized mammals were sampled on Staten Island, New York, United States, to determine H. longicornis host associations and vector potential for a range of human and veterinary pathogens. A total of 97 hosts were sampled and five species of tick (Amblyomma americanum, Dermacentor variabilis, H. longicornis, Ixodes scapularis, Ixodes cookei) were found feeding concurrently on these hosts. Haemaphysalis longicornis was found in the highest proportions compared with other native tick species on raccoons (55.4%), Virginia opossums (28.9%), and white-tailed deer (11.5%). Tissue, blood, and engorged larvae were tested for 17 different pathogens using a nanoscale PCR platform. Infection with five pathogens (Borrelia burgdorferi, Anaplasma phagocytophilum, Rickettsia spp., Mycoplasma haemocanis, and Bartonella spp.) was detected in host samples, but no pathogens were found in any larval samples. These results suggest that although large and medium sized mammals feed large numbers of H. longicornis ticks in the environment, there is presently a low potential for H. longicornis to acquire pathogens from these wildlife hosts.

Bacterial expression of a snake venom metalloproteinase inhibitory protein from the North American opossum (D.virginiana).

A variety of opossum species are resistant to snake venoms due to the presence of antihemorrhagic and antimyotoxic acidic serum glycoproteins that inhibit several toxic venom components. Two virtually identical antihemorrhagic proteins isolated from either the North American opossum (D. virginiana) or the South American big-eared opossum (D. aurita), termed oprin or DM43 respectively, inhibit specific snake venom metalloproteinases (SVMPs). A better understanding of the structure of these proteins may provide useful insight to determine their mechanism of action and for the development of therapeutics against the global health concern of snake-bite envenomation. The aim of this work is to produce a recombinant snake venom metalloproteinase inhibitor (SVMPI) similar to the above opossum proteins in Escherichia coli and determine if this bacterially produced protein inhibits the proteolytic properties of Western Diamondback rattlesnake (C. atrox) venom. The resulting heterologous SVMPI was produced with either a 6-Histidine or maltose binding protein (MBP) affinity tag on either the C-terminus or N-terminus of the protein, respectively. The presence of the solubility enhancing MBP affinity tag resulted in significantly more soluble protein expression. The inhibitory activity was measured using two complementary assays and the MBP labeled SVMPI showed 7-fold less activity as compared to the 6-Histidine labeled SVMPI. Thus, the bacterially derived SVMPI with an unlabeled N-terminus showed high inhibitory activity (IC50 = 4.5 µM). The use of a solubility enhancing MBP fusion protein construct appears to be a productive way to express sufficient quantities of this mammalian protein in E. coli for further study.

Are Virginia opossums really ecological traps for ticks? Groundtruthing laboratory observations.

Virginia opossums (Didelphis virginiana) are a common synanthrope in North America, and serve as host to many species of ectoparasites. Research on captive Virginia opossums estimated that opossums eat, on average, 5500 larval ticks (Acari: Ixodidae) per week. To investigate this apparent preference exhibited by opossums for ingesting ticks, we comprehensively analyzed stomach contents of 32 Virginia opossums from central Illinois. Using a dissecting microscope, we searched the contents exhaustively for ticks and tick body parts, without sieving or pre-rinsing the stomach contents. We did not locate any ticks or tick parts in the stomach contents of Virginia opossums. We also performed a vigorous literature search for corroborating evidence of tick ingestion. Our search revealed 23 manuscripts that describe diet analyses of Virginia opossums, 19 of which were conducted on stomach or digestive tract contents and four of which were scat-based analyses. None of the studies identified ticks in their analyses of diet items. We conclude that ticks are not a preferred diet item for Virginia opossums. Considering that wildlife unconditioned to laboratory conditions may exhibit non-typical behaviors, we recommend that lab-based studies of wildlife behavior be groundtruthed with studies based in natural conditions.

Mesocarnivore landscape use along a gradient of urban, rural, and forest cover.

Mesocarnivores fill a vital role in ecosystems through effects on community health and structure. Anthropogenic-altered landscapes can benefit some species and adversely affect others. For some carnivores, prey availability increases with urbanization, but landscape use can be complicated by interactions among carnivores as well as differing human tolerance of some species. We used camera traps to survey along a gradient of urban, rural, and forest cover to quantify how carnivore landscape use varies among guild members and determine if a species was a human exploiter, adapter, or avoider. Our study was conducted in and around Corvallis, Oregon from April 2018 to February 2019 (11,914 trap nights) using 47 camera trap locations on a gradient from urban to rural. Our focal species were bobcat (Lynx rufus), coyote (Canis latrans), gray fox (Urocyon cinereoargenteus), opossum (Didelphis virginiana), raccoon (Procyon lotor), and striped skunk (Mephitis mephitis). Raccoon and opossum were human exploiters with low use of forest cover and positive association with urban and rural developed areas likely due to human-derived resources as well as some refugia from larger predators. Coyote and gray fox were human adapters with high use of natural habitats while the effects of urbanization ranged from weak to indiscernible. Bobcat and striped skunk appeared to be human avoiders with negative relationship with urban cover and higher landscape use of forest cover. We conducted a diel temporal activity analysis and found mostly nocturnal activity within the guild, but more diurnal activity by larger-bodied predators compared to the smaller species. Although these species coexist as a community in human-dominated landscapes throughout much of North America, the effects of urbanization were not equal across species. Our results, especially for gray fox and striped skunk, are counter to research in other regions, suggesting that mesopredator use of urbanized landscapes can vary depending on the environmental conditions of the study area and management actions are likely to be most effective when decisions are based on locally derived data.

EVIDENCE OF LEPTOSPIRA SEROVARS IN WILDLIFE AND LEPTOSPIRAL DNA IN WATER SOURCES IN A NATURAL AREA IN EAST-CENTRAL ILLINOIS, USA.

We identified seven Leptospira serovars in wildlife and the presence of leptospiral DNA in water sources at a natural area within a fragmented habitat in Illinois, US. These serovars have been implicated in domestic animal and human leptospirosis, a reemerging zoonotic disease, whose reservoirs include wildlife and domestic animals. We live trapped medium-sized mammals (n=351) near building (H-sites) or forest sites (F-sites). Using serology, we evaluated exposure to Leptospira (L. interrogans serovars Autumnalis, Bratislava, Canicola, Icterohaemorrhagiae, Pomona; L. kirschneri serovar Grippotyphosa; L. borgpetersenii serovar Hardjo). Using PCR, we tested for the presence of leptospires in eight water samples (ponds, creeks, and rainwater runoff) collected near trapping sites. We identified antibody titers in raccoons (Procyon lotor; 121/221) and Virginia opossums (Didelphis virginiana; 60/112), but not in feral cats (Felis catus; 0/18). We found significant differences in overall Leptospira seroprevalence between years (P=0.043) and animal's age in 2008 (P=0.005) and 2009 (P=0.003). Serovars Autumnalis, Bratislava, and Grippotyphosa showed significant differences among age groups with the highest seroprevalence in adults. Females had a higher seroprevalence for Icterohaemorragiae in 2008 (P=0.003) and Hardjo in 2009 (P=0.041). Risk of exposure to Leptospira was higher at F-sites compared to H-sites (odds ratio 2.3, 95% confidence interval 1.3-3.9, P=0.002). We captured more animals with titers >1:800 at H-sites, but there was no association between titer levels and capture site. Six of eight water sources were Leptospira-positive; however, there was no correlation between trapping locations of seropositive animals and positive water sources. Natural areas create opportunities for interspecies interactions, favoring leptospires transmission across species. Understanding that Leptospira serovars are present in natural areas is an integral part of the safe human and pet recreational use of these areas. Our study should raise awareness and build on public education designed to prevent disease transmission between species.

Spontaneous neoplasms in captive Virginia opossums ( Didelphis virginiana): a retrospective case series (1989-2014) and review of the literature.

This retrospective project summarizes the types of neoplasms identified in Virginia opossums ( Didelphis virginiana) presented to the University of Tennessee, College of Veterinary Medicine (UTCVM) postmortem service in 1989-2014 and serves as a review of the literature. Of the 85 Virginia opossums identified from the UTCVM case database, there were 17 diagnoses of neoplasia from 12 cases (14%). These cases included 8 females, 2 males, and 2 neutered males. All opossums with known ages (11 of 12) were >2 y old. Pulmonary tumors, specifically minimally invasive or lepidic-predominant adenocarcinomas, were the most common diagnosis and accounted for 53% (9 of 17) of the neoplasms. Additional tumors included acute myeloid leukemia with eosinophil maturation, hepatic hemangiosarcoma, sarcoma (unknown origin), squamous cell carcinoma, disseminated mast cell tumor, trichoblastoma, thyroid adenoma, and an osteoma. These findings serve as a reference for the types of spontaneous neoplasms in Virginia opossums; based on these findings, neoplasia should be considered as a differential in mature captive Virginia opossums.

First molecular evidence of Toxoplasma gondii in opossums (Didelphis virginiana) from Yucatan, Mexico.

Toxoplasma gondii is an obligate intracellular parasite recognized as a causal agent of toxoplasmosis; zoonotic disease endemic in many countries worldwide, including Mexico. Different species of animals participate in the wild cycle infection, including opossums of the species Didelphis virginiana. Thirteen D. virginiana were captured in Yucatan, Mexico. Detection of T. gondii was achieved by Polymerase Chain Reaction, which determined an infection of 76.9% (10/13) in brains. Positive amplicons were sequenced for analysis, this produced results similar to T. gondii with identity and coverage values of 98% and 96-100%, respectively. This study presents the first molecular evidence of the circulation of T. gondii in D. virginiana from Mexico.