Antifungal Synergy: Mechanistic Insights into the R-1-R Peptide and Bidens pilosa Extract as Potent Therapeutics against Candida spp. through Proteomics.

Previous reports have demonstrated that the peptide derived from LfcinB, R-1-R, exhibits anti-Candida activity, which is enhanced when combined with an extract from the Bidens pilosa plant. However, the mechanism of action remains unexplored. In this research, a proteomic study was carried out, followed by a bioinformatic analysis and biological assays in both the SC5314 strain and a fluconazole-resistant isolate of Candida albicans after incubation with R-1-R. The proteomic data revealed that treatment with R-1-R led to the up-regulation of most differentially expressed proteins compared to the controls in both strains. These proteins are primarily involved in membrane and cell wall biosynthesis, membrane transport, oxidative stress response, the mitochondrial respiratory chain, and DNA damage response. Additionally, proteomic analysis of the C. albicans parental strain SC5314 treated with R-1-R combined with an ethanolic extract of B. pilosa was performed. The differentially expressed proteins following this combined treatment were involved in similar functional processes as those treated with the R-1-R peptide alone but were mostly down-regulated (data are available through ProteomeXchange with identifier PXD053558). Biological assays validated the proteomic results, evidencing cell surface damage, reactive oxygen species generation, and decreased mitochondrial membrane potential. These findings provide insights into the complex antifungal mechanisms of the R-1-R peptide and its combination with the B. pilosa extract, potentially informing future studies on natural product derivatives.

Femora nutrient foramina and aerobic capacity in giant extinct xenarthrans.

Nutrient foramina are small openings in the periosteal surface of the mid-shaft region of long bones that traverse the cortical layer and reach the medullary cavity. They are important for the delivery of nutrients and oxygen to bone tissue and are crucial for the repair and remodeling of bones over time. The nutrient foramina in the femur's diaphysis are related to the energetic needs of the femur and have been shown to be related to the maximum metabolic rate (MMR) of taxa. Here, we investigate the relationship between nutrient foramen size and body mass as a proxy to the aerobic capacity of taxa in living and extinct xenarthrans, including living sloths, anteaters, and armadillos, as well as extinct xenarthrans such as glyptodonts, pampatheres, and ground sloths. Seventy femora were sampled, including 20 from extant taxa and 50 from extinct taxa. We obtained the blood flow rate (Q̇) based on foramina area and performed PGLS and phylogenetic ANCOVA in order to explore differences among mammalian groups. Our results show that, among mammals, taxa commonly associated with lower metabolism like living xenarthrans showed relatively smaller foramina, while the foramina of giant extinct xenarthrans like ground sloths and glyptodonts overlapped with non-xenarthran placentals. Consequently, Q̇ estimations indicated aerobic capacities comparable to other placental giant taxa like elephants or some ungulates. Furthermore, the estimation of the MMR for fossil giant taxa showed similar results, with almost all taxa showing high values except for those for which strong semi-arboreal or fossorial habits have been proposed. Moreover, the results are compatible with the diets predicted for extinct taxa, which indicate a strong consumption of grass similar to ungulates and in contrast to the folivorous or insectivorous diets of extant xenarthrans. The ancestral reconstruction of the MMR values indicated a lack of a common pattern for all xenarthrans, strongly supporting the occurrence of low metabolic rates in extant forms due to their particular dietary preferences and arboreal or fossorial habits. Our results highlight the importance of considering different evidence beyond the phylogenetic position of extinct taxa, especially when extinct forms are exceptionally different from their extant relatives. Future studies evaluating the energetic needs of giant extinct xenarthrans should not assume lower metabolic rates for these extinct animals based solely on their phylogenetic position and the observations on their extant relatives.

Predicting the potential distribution and coexistence of Chagas disease vectors in the Americas.

Predicting the potential distribution and coexistence of suitable geographic areas for Chagas disease vectors in the Americas is a crucial task for understanding the eco-epidemiological dynamics of this disease. The potential distribution and coexistence of 3 species-Rhodnius prolixus (Hemiptera: Reduviidae), Cavernicola pilosa (Hemiptera: Reduviidae), and Rhodnius pictipes (Hemiptera: Reduviidae) were modeled. Presence records were obtained and environmental variables were selected based on correlation analysis, Jackknife analysis and knowledge of the biology and natural history of the species. The MaxEnt algorithm included in the kuenm package of R software was used for modeling the potential distribution, and various scenarios of the BAM diagram (Biotic, Abiotic, and Movement variables) were evaluated. The variables contributing to the final models were different for each species. Rhodnius pictipes showed a potential distribution in South America, particularly in Brazil, Bolivia, Peru, Colombia, Venezuela, Guyana, and Suriname. Areas with environmentally suitable conditions for R. prolixus were located in southern Brazil, Peru, Colombia, southern Mexico, Guatemala, El Salvador, and Honduras, whereas for C. pilosa they were in southeastern Brazil, southeastern Central America, Peru, Ecuador, Colombia, Venezuela, Guyana, Suriname, and French Guiana. Co-occurrence analysis revealed distinct patterns in the neotropical region, with some areas indicating the potential distribution of 1 or more species. In Brazil, occurrence and co-occurrence areas were concentrated in the northwest and southeast regions. Overall, this study provides valuable information on the potential distribution and coexistence of vectors, which can inform targeted vector control strategies and contribute to global efforts in combating Chagas disease.

A Bidens pilosa L. Non-Polar Extract Modulates the Polarization of Human Macrophages and Dendritic Cells into an Anti-Inflammatory Phenotype.

Different communities around the world traditionally use Bidens pilosa L. for medicinal purposes, mainly for its anti-inflammatory, antinociceptive, and antioxidant properties; it is used as an ingredient in teas or herbal medicines for the treatment of pain, inflammation, and immunological disorders. Several studies have been conducted that prove the immunomodulatory properties of this plant; however, it is not known whether the immunomodulatory properties of B. pilosa are mediated by its ability to modulate antigen-presenting cells (APCs) such as macrophages (MØs) and dendritic cells (DCs) (through polarization or the maturation state, respectively). Different polar and non-polar extracts and fractions were prepared from the aerial part of B. pilosa. Their cytotoxic and immunomodulatory effects were first tested on human peripheral blood mononuclear cells (PBMCs) and phytohemagglutinin (PHA)-stimulated PBMCs, respectively, via an MTT assay. Then, the non-cytotoxic plant extracts and fractions that showed the highest immunomodulatory activity were selected to evaluate their effects on human MØ polarization and DC maturation (cell surface phenotype and cytokine secretion) through multiparametric flow cytometry. Finally, the chemical compounds of the B. pilosa extract that showed the most significant immunomodulatory effects on human APCs were identified using gas chromatography coupled with mass spectrometry. The petroleum ether extract and the ethyl acetate and hydroalcoholic fractions obtained from B. pilosa showed low cytotoxicity and modulated the PHA-stimulated proliferation of PBMCs. Furthermore, the B. pilosa petroleum ether extract induced M2 polarization or a hybrid M1/M2 phenotype in MØs and a semi-mature status in DCs, regardless of exposure to a maturation stimulus. The immunomodulatory activity of the non-polar (petroleum ether) extract of B. pilosa on human PBMC proliferation, M2 polarization of MØs, and semi-mature status in DCs might be attributed to the low-medium polarity components in the extract, such as phytosterol terpenes and fatty acid esters.

Combining the Peptide RWQWRWQWR and an Ethanolic Extract of Bidens pilosa Enhances the Activity against Sensitive and Resistant Candida albicans and C. auris Strains.

The antifungal activity of palindromic peptide RWQWRWQWR and its derivatives was evaluated against clinical isolates of Candida albicans and C. auris. Also, Bidens pilosa ethanolic extracts of leaves and stem were evaluated. Furthermore, combinations of peptide, extract, and/or fluconazole (FLC) were evaluated. The cytotoxicity of peptides and extracts in erythrocytes and fibroblasts was determined. The original palindromic peptide, some derivative peptides, and the ethanolic extract of leaves of B. pilosa exhibited the highest activity in some of the strains evaluated. Synergy was obtained between the peptide and the FLC against C. auris 435. The combination of the extract and the original palindromic peptide against C. albicans SC5314, C. auris 435, and C. auris 537 decreased the minimal inhibitory concentrations (MICs) by a factor of between 4 and 16. These mixtures induced changes in cell morphology, such as deformations on the cell surface. The results suggest that the combination of RWQWRWQWR and B. pilosa extract is an alternative for enhancing antifungal activity and decreasing cytotoxicity and costs and should be considered to be a promising strategy for treating diseases caused by Candida spp.

Antispasmodic Effect of Valeriana pilosa Root Essential Oil and Potential Mechanisms of Action: Ex Vivo and In Silico Studies.

Infusions of Valeriana pilosa are commonly used in Peruvian folk medicine for treating gastrointestinal disorders. This study aimed to investigate the spasmolytic and antispasmodic effects of Valeriana pilosa essential oil (VPEO) on rat ileum. The basal tone of ileal sections decreased in response to accumulative concentrations of VPEO. Moreover, ileal sections precontracted with acetylcholine (ACh), potassium chloride (KCl), or barium chloride (BaCl2) were relaxed in response to VPEO by a mechanism that depended on atropine, hyoscine butylbromide, solifenacin, and verapamil, but not glibenclamide. The results showed that VPEO produced a relaxant effect by inhibiting muscarinic receptors and blocking calcium channels, with no apparent effect on the opening of potassium channels. In addition, molecular docking was employed to evaluate VPEO constituents that could inhibit intestinal contractile activity. The study showed that α-cubebene, ß-patchoulene, ß-bourbonene, ß-caryophyllene, α-guaiene, γ-muurolene, valencene, eremophyllene, and δ-cadinene displayed the highest docking scores on muscarinic acetylcholine receptors and voltage-gated calcium channels, which may antagonize M2 and/or M3 muscarinic acetylcholine receptors and block voltage-gated calcium channels. In summary, VPEO has both spasmolytic and antispasmodic effects. It may block muscarinic receptors and calcium channels, thus providing a scientific basis for its traditional use for gastrointestinal disorders.

Immunomodulatory Properties of Natural Extracts and Compounds Derived from Bidens pilosa L.: Literature Review.

Bidens pilosa L. has been used in different parts of the world mainly to treat diseases associated with immune response disorders, such as autoimmunity, cancer, allergies, and infectious diseases. The medicinal properties of this plant are attributed to its chemical components. Nevertheless, there is little conclusive evidence that describes the immunomodulatory activity of this plant. In this review, a systematic search was carried out in the PubMed-NLM, EBSCO Host and BVS databases focused on the pre-clinical scientific evidence of the immunomodulatory properties of B. pilosa. A total of 314 articles were found and only 23 were selected. The results show that the compounds or extracts of Bidens modulate the immune cells. This activity was associated with the presence of phenolic compounds and flavonoids that control proliferation, oxidative stress, phagocytosis, and the production of cytokines of different cells. Most of the scientific information analyzed in this paper supports the potential use of B. pilosa mainly as an anti-inflammatory, antioxidant, antitumoral, antidiabetic, and antimicrobial immune response modulator. It is necessary that this biological activity be corroborated through the design of specialized clinical trials that demonstrate the effectiveness in the treatment of autoimmune diseases, chronic inflammation, and infectious diseases. Until now there has only been one clinical trial in phase I and II associated with the anti-inflammatory activity of Bidens in mucositis.

Xenarthrans of the collection of Santiago Roth from the Pampean Region of Argentina (Pleistocene), in Zurich, Switzerland.

The present work concerns xenarthrans from the collection of Santiago (Kaspar Jakob) Roth (1850-1924) housed at the Palaeontological Institute and Museum of the University of Zurich, one of the most important collections of Pleistocene mammals from Argentina in Europe. Roth was a paleontologist originally from Switzerland who prospected and collected a large amount of Pleistocene megafauna of the Pampean Region of Argentina. The xenarthrans are the main representatives of this collection in Zurich, with 150 specimens. Since 1920, this material has not been revised and is under studied. The present investigation corresponds to a taxonomic revision resulting in 114 reassignments, leading to document xenarthran diversity and discuss their paleoecologies. The high diversity reflects the paleoecology of the Pampean Region during the Pleistocene, with the various abiotic events that impacted the paleoenvironment of this region. Within the Cingulata, the Pampean Region fauna was probably dominated by glyptodonts with a high representation of Glyptodontinae and Neosclerocalyptinae while within the sloths the highest diversity and abundance is found in the Mylodontinae and Scelidotheriinae. These four clades represent both species with high ecological tolerance (e.g., Glyptodon munizi; Catonyx tarijensis) and ecologically highly specialized species (e.g., Neosclerocalyptus paskoensis; Scelidotherium leptocephalum). The presence of such ecological diversity underlines the status of the Pampean Region as a major interest for paleoecological and paleoenvironmental reconstruction. Supplementary Information: The online version contains supplementary material available at 10.1186/s13358-023-00265-7.

El presente trabajo se refiere a los xenartros de la colección de Santiago (Kaspar Jakob) Roth (1850-1924) depositada en el Instituto y Museo Paleontológico de la Universidad de Zurich, una de las más importantes colecciones de mamíferos del Pleistoceno de Argentina en Europa. Roth fue un paleontólogo originario de Suiza que prospectó y coleccionó una gran cantidad de megafauna del Pleistoceno de la Región Pampeana de Argentina. Los xenartros son los principales representantes de esta colección en Zurich, con 150 ejemplares. Desde 1920, este material no ha sido revisado y está poco estudiado. La presente investigación corresponde a una revisión taxonómica que ha dado lugar a 110 reasignaciones, lo que permite documentar la diversidad de los xenartros y discutir aspectos paleoecológicos. La alta diversidad refleja la paleoecología de la Región Pampeana durante el Pleistoceno en asociación con los cambios climáticos que han impactado en el paleoambiente de esta región. Entre los Cingulata, la fauna de la Región Pampeana estuvo probablemente dominada por los gliptodontes con una alta representación de Glyptodontinae y Neosclerocalyptinae, mientras que entre los perezosos la mayor diversidad y abundancia se encuentra en los Mylodontinae y Scelidotherinae. Estos cuatro clados representan tanto especies con alta tolerancia ecológica (por ejemplo, Glyptodon munizi; Catonyx tarijensis) como otras muy especializadas ecológicamente (por ejemplo, Neosclerocalyptus paskoensis; Scelidotherium leptocephalum). La presencia de tal diversidad ecológica subraya el estatus de la Región Pampeana como de gran interés para la reconstrucción paleoecológica y paleoambiental.

Ex Situ Approaches for the Conservation of Genetic Resources in the Superorder Xenarthra.

Xenarthra-a superorder of placental mammals endemic to the Neotropics-is represented by armadillos, anteaters, and sloths. Considering their long history in the Americas, extant xenarthrans represent an important group for understanding the impact of past environmental changes on species diversification and serve key ecological functions as ecosystem engineers. Unfortunately, most wild xenarthran populations are at risk, due primarily to anthropogenic activities, necessitating urgent conservation efforts. Moreover, the paucity of information on some species has rendered population estimation and, consequently, conservation management challenging. In addition, relatively few groups are researching this superorder, perhaps because fieldwork with armadillos, anteaters, or sloths and their captive care are challenging tasks. Nevertheless, dedicated research and efforts to ensure the long-term conservation of these animals are deemed essential. In this context, cryobanks are a practical approach for breeding and maintaining genetic diversity in wildlife, and they are important tools for assisting and improving both ex situ and in situ conservation strategies. Therefore, cryopreservation of biological resources may be a promising strategy for conserving xenarthrans. Specifically, semen cryopreservation, which has already been applied in some species, may be the most effective strategy for this group. The present article provides an overview of ex situ conservation of xenarthrans, which will contribute to the development and implementation of additional strategies for protecting these unique mammals.

Threats to health and conservation of free-living sloths (Bradypus and Choloepus) under anthropic influence in the city of Manaus, Amazonas state, Brazil / Ameaças à saúde e conservação de preguiças (Bradypus e Choloepus) de vida livre sob influência antrópica em Manaus, Estado do Amazonas, Brasil

Abstract Anthropogenic disturbances affecting forest areas can increase disease prevalence and susceptibility in several species of arboreal mammals, such as sloths. Thus, this study aimed to evaluate the most common conditions in body systems of free-ranging sloths admitted at the Wildlife Triage and Rehabilitation Center of Amazonas of the Institute of Environment and Renewable Natural Resources. A total of 227 individuals (139 retrospective cases from 2015 to 2019, and 88 prospective cases from July 2020 to July 2021) were evaluated over a six-year period. Cases from the genus Bradypus showed involvement of the following body systems: 44% respiratory, 29% integumentary, 15% musculoskeletal, 5% digestive, 3% visual, 2% auditory, 1% circulatory and 1% genitourinary, while that distribution in the genus Choloepus was: 39% musculoskeletal, 27% integumentary, 19% respiratory, 9% digestive, 3% circulatory, 1% visual, 1% auditory and 1% genitourinary. The results reveal significant differences between the condition detected and the genus (Bradypus and Choloepus), age and case outcome. These results can provide data for future investigations of sloth diseases, confirming lesions, as well as motivating and suggesting adequate management methods.

Resumo Distúrbios antrópicos que afetam áreas florestais podem aumentar a prevalência de afecções e a suscetibilidade às doenças em diversas espécies de mamíferos arborícolas, como as preguiças. Desta forma, este estudo teve como objetivo avaliar as condições mais comuns nos sistemas orgânicos de preguiças de vida livre admitidas no Centro de Triagem e Reabilitação de Animais Silvestres do Amazonas. Um total de 227 indivíduos (139 casos retrospectivos de 2015 a 2019 e 88 casos prospectivos de julho de 2020 a julho de 2021) foram avaliados durante um período de seis anos. Para o gênero Bradypus foram diagnosticadas 44% de afecções do sistema respiratório, 29% tegumentar, 15% musculoesquelético, 5% digestório, 3% visual, 2% auditivo, 1% circulatório e 1% genitourinário. Por sua vez, para o gênero Choloepus foram detectadas 39% de afecções do sistema musculoesquelético, 27% tegumentar, 19% do sistema respiratório, 19% do sistema respiratório, 9% digestório, 3% circulatório, 1% visual, 1% auditivo e 1% genitourinário. Os resultados mostraram diferenças significativas na prevalência da afecção e gênero das preguiças (Bradypus e Choloepus), a faixa etária e desfecho do caso. Esses resultados podem fornecer subsídios para investigar as doenças de preguiças, confirmando afecções, motivo e sugerir métodos adequados de manejo.

Phytochemical study of the plant species Bidens pilosa L. (Asteraceae) and Croton floccosus (Euphorbiaceae).

Background: Given the chemical richness of medicinal plants ( Bidens pilosa L. and Croton floccosus) in Ecuador, they are considered the natural source of numerous medicines. Methods: The leaves were dried at 40°C and 50°C and the extracts were characterized by means of phytochemical screening, verifying the presence of secondary metabolites such as alkaloids, reducing sugars, phenols, flavonoids, tannins and saponins. Three extraction processes were carried out, with two solvents of different polarities: hexane and ethanol. The extraction methods that were applied to the leaves of the plants were Soxhlet, ultrasonic bath and maceration, the latter two at room temperature and Soxhlet at the boiling temperature of the solvent. Determination of the total content of phenols and flavonoids is carried out using the Follin-Ciocalteau colorimetric reaction, Quercetin standard, Aluminum Chloride solution measured with a UV-Vis spectrophotometer. The antioxidant activity was performed with the DPPH radical and measured with the same equipment. Results: The highest content of total phenols obtained by employing the Soxhlet method for extraction when the material was dried at 50°C was 48.609 ± 0.370 mg GAE/g of dry sample for Bidens pilosa L. while in the case of Croton floccosus it was 128.212 ± 0.601 mg GAE/g of dry sample obtained from the extraction by means of maceration. Finally, the antioxidant activity against the 1.1-diphenyl-2-picryl-hydrazyl radical was determined, and it was found that the Bidens pilosa L. species performed better and responded better to the test, with an IC 50 value of 239.33 µg/mL, than Croton floccosus (IC 50 of 644.125 µg/mL). Conclusions: The following preliminary phytochemical study of the Bidens pilosa L. and Croton floccosus plants provided important information on the content of secondary metabolites and response to the DPPH radical reported for the first time in Ecuador, which may be future use for medicinal application.

Video Validation of Tri-Axial Accelerometer for Monitoring Zoo-Housed Tamandua tetradactyla Activity Patterns in Response to Changes in Husbandry Conditions.

Accelerometers are a technology that is increasingly used in the evaluation of animal behaviour. A tri-axial accelerometer attached to a vest was used on Tamandua tetradactyla individuals (n = 10) at Biodiversity Park. First, the influence of using a vest on the animals' behaviour was evaluated (ABA-type: A1 and A2, without a vest; B, with a vest; each stage lasted 24 h), and no changes were detected. Second, their behaviour was monitored using videos and the accelerometer simultaneously (experimental room, 20 min). The observed behaviours were correlated with the accelerometer data, and summary measures (X, Y and Z axes) were obtained. Additionally, the overall dynamic body acceleration was calculated, determining a threshold to discriminate activity/inactivity events (variance = 0.0055). Then, based on a 24 h complementary test (video sampling every 5 min), the sensitivity (85.91%) and precision (100%) of the accelerometer were assessed. Animals were exposed to an ABA-type experimental design: A1 and A2: complex enclosure; B: decreased complexity (each stage lasted 24 h). An increase in total activity (%) was revealed using the accelerometer (26.15 ± 1.50, 29.29 ± 2.25, and 35.36 ± 3.15, respectively). Similar activity levels were detected using video analysis. The results demonstrate that the use of the accelerometer is reliable to determine the activity. Considering that the zoo-housed lesser anteaters exhibit a cathemeral activity pattern, this study contributes to easily monitoring their activities and responses to different management procedures supporting welfare programs, as well as ex situ conservation.

Hepatocellular Carcinoma in a Free-Ranging Three-Toed Sloth (Bradypus variegatus).

The increasing interest of tumors in wildlife is important for biodiversity conservation and for monitoring environmental agents and/or contaminants with potential impact on human health. Here we described the occurrence of hepatocellular carcinoma (HCC) in noncirrhotic liver of a free-ranging three-toed sloth (Bradypus variegatus) from the Atlantic Forest biome in Brazil. The HCC showed a moderate mononuclear inflammatory infiltrate within the tumor tissue but with no inflammation and fibrosis in the adjacent liver tissue. Upon immunohistochemistry, neoplastic cells were diffusely positive for HepPar-1 and glutamine-synthetase presenting an irregular and random immunostaining pattern; ß-catenin was positive in the cytoplasmic membrane of malignant hepatocytes; and cytokeratin 19 immunostaining was restricted to bile duct epithelial cells. The liver tissue was negative for HBV-like and HCV-like viruses assessed by molecular tests. The potential similarity of pathogenesis may reinforce the need for research on environmental and/or infectious agents associated with HCC that may contribute to the understanding of cancer in wildlife.

Valeriana pilosa Roots Essential Oil: Chemical Composition, Antioxidant Activities, and Molecular Docking Studies on Enzymes Involved in Redox Biological Processes.

Valeriana pilosa is usually employed in Peruvian folk medicine in the form of infusion to treat stomach pain, and has antispasmodic, relaxing, sleep-promoting, and sedative properties, as well as is an anti-inflammatory. In this study, Valeriana pilosa essential oil (VPEO) was obtained by hydrodistillation, analyzed by GC and GC/MS, and 47 compounds were identified. Major oil components were α-patchoulene (5.8%), α-humulene (6.1%), seychellene (7.6%), and patchoulol (20.8%). Furthermore, we assessed the in vitro antioxidant activities, molecular docking, and Ligand Efficiency studies on enzymes involved in cellular redox pathways such as CYP2C9, catalase, superoxide dismutase, and xanthine oxidase. Essential oil antioxidant activities were assessed by FRAP, ABTS•+, and DPPH• radical scavenging activity. VPEO displays high antioxidant activity as compared to essential oils of Valeriana jatamansi and Valeriana officinalis oil roots. In addition, molecular docking and ADMET prediction was employed to compare the absorption, metabolism, and toxicity properties of Valeriana pilosa compounds. In the molecular docking studies, limonene, p-cimene, carvone, α-cubebene, cyclosativene, α-guaiene, allo-aromadendrene, valencene, and eremophyllene were the compounds with the best docking score on CYP2C9 and xanthine oxidase. Thus, volatile components of Valeriana pilosa could be associated with the detected antioxidant activity, acting as putative inhibitors of CYP2C9 and xanthine oxidase.

Hematologic and biochemical reference intervals of brown-throated sloths (Bradypus variegatus).

The brown-throated sloth, Bradypus variegatus, is a common species endemic to South and Central America. Nonetheless, maintaining these animals in captivity can be challenging, and very few institutions manage to do so. The São Paulo Zoological Park Foundation is in a remnant of the Atlantic rainforest in the middle of São Paulo, the largest city in Brazil. This forest fragment has a population of B. variegatus that is geographically isolated and yet to be studied. Assessing the health status of individuals remains difficult due to the lack of reference intervals (RIs) for hematologic and biochemical variables for this species. We aimed to establish hematologic and biochemical RIs in a population of wild B. variegatus living in the largest remnant of Atlantic rainforest in São Paulo city, Brazil. Blood samples from 25 individuals of wild B. variegatus were collected and analyzed for 20 hematologic and 21 biochemical variables, using standard laboratory techniques. Each variable was statistically analyzed according to the American Society for Veterinary Clinical Pathology guidelines. The results obtained for each variable were statically analyzed, making it possible to obtain descriptive statistics for all hematologic and biochemical variables. RIs were determined for 16 hematologic variables. During the microscopic analysis, we observed anisocytosis, polychromatophils, Howell-Jolly bodies, macroplatelets, and reactive lymphocytes. The RIs and descriptive statistics described here establish important baseline numbers that could be essential for the management and treatment of both captive and wild B. variegatus sloths.

The eye of the giant anteater (Myrmecophaga tridactyla): Ophthalmic examination findings and diagnostic tests.

OBJECTIVE: To describe ophthalmic examination and diagnostic values for Schirmer tear test (STT), intraocular pressure (IOP), corneal horizontal diameter (CHD), palpebral fissure length (PFL), fundoscopy, and palpebral conjunctiva microbiota from healthy giant anteaters. ANIMALS STUDIED: Twelve giant anteaters (Myrmecophaga tridactyla), 11 adults and one juvenile, five males and seven females. PROCEDURES: The animals were submitted to general anesthesia and ophthalmic evaluation with portable slit-lamp biomicroscope, Finoff transilluminator, and fundoscopy, as well as STT, bacterial culture from palpebral conjunctiva, rebound tonometry IOP, and measurement of PFL and CHD. Data compiled were analyzed with ANOVA and Tukey tests. RESULTS: The results (mean ± standard deviation) were as follows: STT 8.04 ± 6.21 mm/min; IOP 10.92 ± 2.45 mmHg; PFL 0.75 ± 0.11 cm; CHD 0.96 ± 0.10 cm. Out of the 24 eyes swab samples for bacterial culture, 17 were positive, with three genera of Gram-positive bacteria identified Staphylococcus spp., Bacillus sp., and Corynebacterium sp. Gram-negative bacteria were not isolated from any of the samples. CONCLUSIONS: As conservation work in this vulnerable species continues, this report on basic ophthalmic examination and diagnostic parameters will be helpful improve their treatment and care. More ophthalmic studies are encouraged in animals within the Pilosa order.

A Novel Host of an Emerging Disease: SARS-CoV-2 Infection in a Giant Anteater (Myrmecophaga tridactyla) Kept Under Clinical Care in Brazil.

A young male free-ranging giant anteater (Myrmecophaga tridactyla) was found with paralysis of pelvic limbs on a highway and kept under human care. Radiographs confirmed multiple incomplete fractures in the thoracolumbar vertebrae. Due to the poor prognosis, euthanasia was chosen. The infection was established by viral SARS-CoV-2 RNA detection in the rectal swab, spleen and kidney samples. Immunohistochemistry detected the viral nucleocapsid protein in sections of the lungs, liver, spleen, lymph nodes, and large intestine sections, and spike protein antigen in the lung tissue. Pilosa order species should be included as potential hosts of natural infection of SARS-CoV-2.

Acarofauna present in organic strawberry fields and associated weed species in southern Brazil.

The presence of weeds in the margins of strawberry crops can enhance populations of predatory mites as a measure to support conservation biological control. The aims of this study were (i) to assess the composition of the acarofauna associated with strawberries and the accompanying herbaceous plants in an organic farming system, and (ii) to evaluate the possible relationships between phytophagous and predatory mites occurring in this system. Strawberry leaves and whole plants of weeds were sampled biweekly from August 2014 to February 2015 in Lapa (Paraná, Brazil). In total, 23 weed species belonging to 10 families were identified; 3768 mite individuals (from 15 families and 4 suborders) were recovered, 77% on strawberries and 23% on weeds. Abundance of predatory mites on weeds was greater than on strawberry cultivars. On strawberries, the most abundant family was Tetranychidae (84%) followed by Phytoseiidae (11.6%). In total, 16 predatory mite species from the Phytoseiidae family were identified, 13 of them occurring on strawberry leaflets. Typholodromalus aripo, Neoseiulus californicus and Typhlodromips mangleae were the most abundant mite species on strawberry leaves. On weeds, most individuals were predatory mites (59%), whereas phytophagous mites represented 17.2%. The most abundant family was Phytoseiidae (36.4%). On weeds, the phytoseiid mite T. aripo was the most abundant species, representing 34.7%. Besides being found on strawberry leaflets, T. aripo was associated with 15 weed species. Among the weeds, Bidens pilosa showed the highest values of the Shannon index (1.97), Margalef index (3.04), and Pielou's evenness index (0.95). This study emphasizes the importance of surrounding weeds as a shelter for beneficial mitefauna in strawberry fields, likely contributing to enhance conservation biological control.