Naturally Acquired Rabies in White-Eared Opossum, Brazil.

Opossums are considered resistant to rabies. Nonhematophagous bats are reservoirs of rabies in urban areas of South America. We analyzed bats and opossums tested for rabies during 2021 in a highly urbanized city in Brazil to understand spillover in an urban setting. Wildlife surveillance is necessary to prevent rabies in humans and domestic animals.

Morphophysiological alterations in fruit-eating bats after oral exposure to deltamethrin.

Deltamethrin (DTM) is a synthetic pyrethroid widely used in the cultivation and management of several crops due to its insecticidal action. Application to crops of pyrethroids such as DTM can result in the exposure of water and fruit consumed by fruit bats having a high pyrethroid content which may be harmful. Therefore the objective of this study was to evaluate the effects of short-term oral exposure of the fruit-eating bats (Artibeus lituratus) to two concentrations of DTM (0.02 and 0.04 mg/kg of papaya) on histopathology of the intestine, liver and kidney. The intestine of the animals exposed to both concentrations showed inflammatory infiltrate, degeneration, necrosis and goblet cell hyperplasia as the most frequent pathologies. Besides, the acid mucins showed an increase in the frequency of non-viable cells. The liver showed hepatocyte vacuolizatio and nuclear enlargement, as well as inflammatory infiltrate and steatosis. The kidneys of the exposed animals showed and inflammatory infiltrate, benign nephrosclerosis, vacuolization and necrosis. Also, DTM reduced nitric oxide synthesis, decreased glomerular diameter and increased glycogen percentage in the proximal tubules. Our results suggest that acute exposure to DTM at low concentrations has the potential to induce pronounced histopathological changes in vital organs, such as intestine, liver and kidney of fruit-eating bats.

Interaction between MHC diversity and constitution, gut microbiota and Astrovirus infections in a neotropical bat.

Astroviruses (AstVs) infect numerous mammalian species including reservoirs such as bats. Peptides encoded by the genes of the highly polymorphic Major Histocompatibility Complex (MHC) form the first line of host defence against pathogens. Aside from direct involvement in mounting adaptive immune responses, MHC class II genes are hypothesized to regulate gut commensal diversity and shape the production of immune-modulatory substances by microbes, indirectly affecting host susceptibility. Despite initial empirical evidence for the link between host MHC and the microbiota, associations among these factors remain largely unknown. To fill this gap, we examined MHC allelic diversity and constitution, the gut bacterial community and abundance pattern of a wild population of a neotropical bat (Artibeus jamaicensis) challenged by AstV infections. First, we show an age-dependent relationship between the host MHC class II diversity and constitution and the gut microbiota in AstV-uninfected bats. Crucially, these associations changed in AstV-infected bats. Additionally, we identify changes in the abundance of specific bacterial taxa linked to the presence of certain MHC supertypes and AstV infection. We suggest changes in the microbiota to be either a result of AstV infection or the MHC-mediated modulation of microbial communities. The latter could subsequently affect microbe-mediated immunity and resistance against AstV infection. Our results emphasize that the reciprocal nature of host immune genetics, gut microbial diversity and pathogen infection require attention, which are particularly important given their repercussions for disease susceptibility and severity in wild animal populations with a history of zoonotic spillover and frequent human contact.

Bats actively modulate membrane compliance to control camber and reduce drag.

Bat wing skin is exceptionally compliant and cambers significantly during flight. Plagiopatagiales proprii, arrays of small muscles embedded in the armwing membrane, are activated during flight and are hypothesized to modulate membrane tension. We examined the function of these muscles using Jamaican fruit bats, Artibeus jamaicensis. When these muscles were paralyzed using botulinum toxin, the bats preferred flight speed decreased and they were unable to fly at very low speeds. Paralysis of the plagiopatagiales also resulted in increased armwing camber consistent with a hypothesized role of modulating aeroelastic interactions. Other compensatory kinematics included increased downstroke angle and increased wingbeat amplitude. These results are consistent with the bats experiencing increased drag and flight power costs associated with the loss of wing-membrane control. Our results indicate that A. jamaicensis likely always employ their wing membrane muscles during sustained flight to control camber and to enhance flight efficiency over a wide flight envelope.

SURGICAL ORCHIECTOMY IN FRUIT BATS-DESCRIPTION OF TWO TECHNIQUES IN THE RUWENZORI LONG-HAIRED FRUIT BAT (ROUSETTUS LANOSUS) AND THE JAMAICAN FRUIT-EATING BAT (ARTIBEUS JAMAICENSIS).

Contraception is needed to prevent overpopulation and inbreeding in highly fecund captive bat colonies. Reports on surgical contraception in bats are limited. The objective of this study was to describe surgical castration techniques in a megachiropteran and a microchiropteran fruit bat species. Open orchiectomy by transfixing ligation of the spermatic cord was performed in 14 Ruwenzori long-haired fruit bats (Rousettus lanosus) (RL-LIG), and orchiectomy with radiosurgery alone was performed in 125 Jamaican fruit-eating bats (Artibeus jamaicensis) (AJ-RS) and one Ruwenzori bat (RL-RS). The surgical techniques were not compared in different species with the exception of the one Ruwenzori bat operated with RS. Anesthesia was induced and maintained with isoflurane in oxygen via facemask. Preoperatively, all bats received butorphanol and subcutaneous fluids. Meloxicam was administered postoperatively for the RL-LIG. For the RL-LIG, anesthesia lasted 49 ± 15 min (mean ± SD) with a total surgery time of 26 ± 12 min. In comparison, the RS was considerably shorter, lasting 10 ± 3 min for anesthesia and 5 ± 2 min for surgery. Complications were rare, with a morbidity rate of 6.7% with the RL-LIG (prolonged recovery [n = 1]) and of 4.8% with the RS (dyspnea [n = 3], hemorrhage [n = 2], and prolonged recovery [n = 1]). One of the cases of hemorrhage was in the single Ruwenzori bat castrated using the RS technique. Mortality rate was 1.6% (n = 2) with the RS. No mortality occurred with the RL-LIG. In conclusion, using radiosurgery alone appears to be a safe and rapid surgical technique in smaller species of fruit bats. For larger species, such as the Ruwenzori fruit bats, ligation or use of an advanced vessel sealing system is recommended.

Dietary Exposure to Tebuconazole Affects Testicular and Epididymal Histomorphometry in Frugivorous Bats.

This study evaluated the effects of a commercially recommended concentration (1 mL/L) of a fungicide tebuconazole (TBZ) on testicular and epididymal histomorphometry of Artibeus lituratus, following 7 and 30-day oral exposure. TBZ30 bats showed a reduction in the percentage of tubules and seminiferous epithelium, as well as a decrease in tubule and epithelium somatic indexes, and tubular diameter. Inversely, these animals showed increased percentage of intertubular compartment, Leydig cells and blood vessels. The volume of Leydig cells and their number per gram of testis also increased in TBZ30 bats. Alterations in epididymal morphometry were observed in all regions of the organ, with increase of ductal diameter in both exposure times. These results indicate that exposure to low concentration of TBZ resulted in testicular and epididymal morphometric changes in fruit bats, mainly at 30-day exposure, suggesting that functional alterations might be occurring in these organs and impacting reproductive capacity.

Characterization of the myoepithelial cells in the major salivary glands of the fruit bat Artibeus jamaicensis.

Bats constitute one of the most numerous mammalian species. Bats have a wide range of dietary habits and include carnivorous, haematophagous, insectivorous, frugivorous and nectivorous species. The salivary glands of these species have been of particular research interest due to their structural variability among chiropterans with different types of diets. Myoepithelial cells (MECs), which support and facilitate the expulsion of saliva from the secretory portions of salivary glands, are very important for their function; however, this cell type has not been extensively studied in the salivary glands of bats. In this study, we characterized the MECs in the major salivary glands of the fruit bat Artibeus jamaicensis. Herein, we describe the morphology of the parotid, submandibular and sublingual glands of A. jamaicensis at the light- and electro-microscopic level and the distribution of MECs in these glands, as defined by their expression of smooth-muscle markers such as α-smooth muscle actin (SMAα) and desmin, and of epithelial cell markers, such as KRT14. We found that the anatomical locations of the major salivary glands in this bat species are similar to those of humans, except that the bat sublingual gland appears to be unique, extending to join the contralateral homologous gland. Morphologically, the parotid gland has the characteristics of a mixed-secretory gland, whereas the submandibular and sublingual glands were identified as mucous-secretory glands. MECs positive for SMAα, KRT14 and desmin were found in all of the structural components of the three glands, except in their excretory ducts. Desmin is expressed at a lower level in the parotid gland than in the other glands. Our results suggest that the major salivary glands of A. jamaicensis, although anatomically and structurally similar to those of humans, play different physiological roles that can be attributed to the dietary habits of this species.

Low, Chronic Exposure to Endosulfan Induces Bioaccumulation and Decreased Carcass Total Fatty Acids in Neotropical Fruit Bats.

We investigated the effects of the insecticide endosulfan on energy metabolism and its possible accumulation in fruit bats. Adult male bats (Artibeus lituratus) were exposed for 35 days, when they were offered fruit treated with endosulfan (E) and adhesive spreader (AS) in the following concentrations (g/L): 0.0; 0.0 (Control), 0.0; 0.015 (AS), 1.05; 0.015 (E1), 2.1; 0.015 (E2). Concentrations used were those recommended by the manufacturer for fruit crop application (E1) or twice this value (E2). E1 bats showed decreased plasma glucose concentration. Carcass fatty acids were decreased in E1 and E2 bats. Endosulfan bioaccumulation was observed in both liver and adipose tissues from E1 and E2 bats. These results indicate that the chronic exposure of fruit bats to environmentally relevant concentrations of endosulfan can lead to significant bioaccumulation beyond control and also decreased fatty acid content, which may impair the health of this important seed disperser in neotropical forests.

Novel Waddlia Intracellular Bacterium in Artibeus intermedius Fruit Bats, Mexico.

An intracellular bacterium was isolated from fruit bats (Artibeus intermedius) in Cocoyoc, Mexico. The bacterium caused severe lesions in the lungs and spleens of bats and intracytoplasmic vacuoles in cell cultures. Sequence analyses showed it is related to Waddlia spp. (order Chlamydiales). We propose to call this bacterium Waddlia cocoyoc.

Postnatal ontogeny of the cochlea and flight ability in Jamaican fruit bats (Phyllostomidae) with implications for the evolution of echolocation.

Recent evidence has shown that the developmental emergence of echolocation calls in young bats follow an independent developmental pathway from other vocalizations and that adult-like echolocation call structure significantly precedes flight ability. These data in combination with new insights into the echolocation ability of some shrews suggest that the evolution of echolocation in bats may involve inheritance of a primitive sonar system that was modified to its current state, rather than the ad hoc evolution of echolocation in the earliest bats. Because the cochlea is crucial in the sensation of echoes returning from sonar pulses, we tracked changes in cochlear morphology during development that included the basilar membrane (BM) and secondary spiral lamina (SSL) along the length of the cochlea in relation to stages of flight ability in young bats. Our data show that the morphological prerequisite for sonar sensitivity of the cochlea significantly precedes the onset of flight in young bats and, in fact, development of this prerequisite is complete before parturition. In addition, there were no discernible changes in cochlear morphology with stages of flight development, demonstrating temporal asymmetry between the development of morphology associated with echo-pulse return sensitivity and volancy. These data further corroborate and support the hypothesis that adaptations for sonar and echolocation evolved before flight in mammals.

High paracellular nutrient absorption in intact bats is associated with high paracellular permeability in perfused intestinal segments.

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular mechanisms. Based on a few previous studies, the capacity for paracellular nutrient absorption seems greater in flying mammals than in nonflying mammals, but there has been little investigation of the mechanisms driving this difference. Therefore, we studied three species each of bats (Artibeus lituratus, Sturnira lilium and Carollia perspicillata) and nonflying mammals (Akodon montensis, Mus musculus and Rattus norvegicus). Using standard pharmacokinetic techniques in intact animals, we confirmed the greater paracellular nutrient absorption in the fliers, comparing one species in each group. Then we conducted in situ intestinal perfusions on individuals of all species. In both approaches, we measured the absorption of 3OMD-glucose, a nonmetabolizable glucose analog absorbed both paracellularly and transcellularly, as well as L-arabinose, which has no mediated transport. Fractional absorption of L-arabinose was three times higher in the bat (S. lilium: 1.2±0.24) than in the rodent (A. montensis: 0.35±0.04), whereas fractional absorption of 3OMD-glucose was complete in both species (1.46±0.4 and 0.97±0.12, respectively). In agreement, bats exhibited two to 12 times higher l-arabinose clearance per square centimeter nominal surface area than rodents in intestinal perfusions. Using L-arabinose, we estimated that the contribution of the paracellular pathway to total glucose absorption was higher in all three bats (109-137%) than in the rodents (13-39%). These findings contribute to an emerging picture that reliance on the paracellular pathway for nutrient absorption is much greater in bats relative to nonflying mammals and that this difference is driven by differences in intestinal permeability to nutrient-sized molecules.

Phylogeography of the dark fruit-eating bat Artibeus obscurus in the Brazilian Amazon.

Artibeus obscurus (Mammalia: Chiroptera) is endemic to South America, being found in at least 18 Brazilian states. Recent studies revealed that different populations of this genus present distinct phylogeographic patterns; however, very little is known on the population genetics structure of A. obscurus in the Amazon rainforest. Here, using a fragment (1010bp) of the mitochondrial gene cytochrome b from 87 samples, we investigated patterns of genetic divergence among populations of A. obscurus from different locations in the Brazilian Amazon rainforest and compared them with other Brazilian and South American regions. Analysis of molecular variance (AMOVA), fixation index (Fst) analysis, and phylogeographic patterns showed divergence between two major monophyletic groups, each one corresponding to a geographic region associated with the Atlantic and Amazon forest biomes. The Atlantic forest clusters formed a monophyletic group with a high bootstrap support and a fragmented distribution that follows the pattern predicted by the Refuge Theory. On the other hand, a different scenario was observed for the Amazon forest, where no fragmentation was identified. The AMOVA results revealed a significant geographic heterogeneity in the distribution of genetic variation, with 70% found within populations across the studied populations (Fst values ranging from 0.05864 to 0.09673; φST = 0.55). The intrapopulational analysis revealed that one population (Bragança) showed significant evidence of population expansion, with the formation of 2 distinct phylogroups, suggesting the occurrence of a subspecies or at least a different population in this region. These results also suggest considerable heterogeneity for A. obscurus in the Amazon region.

A Five-Year Study on Infestation and Abundance of Bat Flies (Hippoboscoidea: Streblidae) Under Severe Dry Season Conditions in the Tropical Dry Forest of Yucatan, Mexico.

In Mexico, few studies have explored how environmental conditions in tropical dry forests (TDF) influence bat fly load even though, according to climate change scenarios, this ecosystem will experience a drier and warmer climate. Such an extension of the dry season in these ecosystems could have dramatic consequences for biodiversity, particularly in regions with plains where animals do not have elevational climate shifts. The present study therefore evaluates the effect of prevailing environmental conditions during 2015-2019, as well as host body conditions, on the infestation and abundance of bat-specific ectoparasites and the composition and bat fly load in the dry season of a TDF in Yucatan. Since Yucatan has an essentially flat and low-lying topography, organisms cannot escape from the predicted extreme conditions with elevational shifts. This region is therefore an excellent location for assessment of the potential effects of warming. We collected 270 bat flies from 12 species. Three streblid species (Nycterophilia parnelli Wenzel, Trichobius johnsonae Wenzel, and Trichobius sparsus Kessel) are new records for Yucatan. Our overview of the dry season bat ectoparasite loads reveals low values of richness and prevalence, but high aggregation. Our models detected significant differences in ectoparasite infestation and abundance over the years, but the environmental and body host condition variables were unrelated to these. We report that pregnant females are parasitized to a greater extent by bat flies during the dry season, which generally represents the season of most significant nutritional stress.

Antigenic and molecular evidence of Brucella sp.-associated epididymo-orchitis in frugivorous (Artibeus lituratus) and nectarivorous (Glossophaga soricina) bats in Brazil.

This study included 47 free-ranging bats from the State of Minas Gerais, Brazil. Six bats (12.8%) had genital inflammatory lesions, and two of them (one Artibeus lituratus and one Glossophaga soricina, a frugivorous and a nectarivorous, respectively) were diagnosed with Brucella sp. infection through PCR, and antigens in intralesional macrophages were detected using immunohistochemistry.

Different but Not Unique: Deciphering the Immunity of the Jamaican Fruit Bat by Studying Its Viriome.

A specialized and fine-tuned immune response of bats upon infection with viruses is believed to provide the basis for a "friendly" coexistence with these pathogens, which are often lethal for humans and other mammals. First insights into the immunity of bats suggest that bats have evolved to possess their own strategies to cope with viral infections. Yet, the molecular details for this innocuous coexistence remain poorly described and bat infection models are the key to unveiling these secrets. In Jamaican fruit bats (Artibeus jamaicensis), a New World bat species, infection experiments with its (putative) natural viral pathogens Tacaribe virus (TCRV), rabies virus (RABV), and the bat influenza A virus (IAV) H18N11, have contributed to an accurate, though still incomplete, representation of the bat-imposed immunity. Surprisingly, though many aspects of their innate and adaptive immune responses differ from that of the human immune response, such as a contraction of the IFN locus and reduction in the number of immunoglobulin subclasses, variations could also be observed between Jamaican fruit bats and other bat species.

The Kidney-Associated Microbiome of Wild-Caught Artibeus spp. in Grenada, West Indies.

Bats are capable of asymptomatically carrying a diverse number of microorganisms, including human pathogens, due to their unique immune system. Because of the close contact between bats and humans, there is a possibility for interspecies transmission and consequential disease outbreaks. Herein, high-throughput sequencing was used to determine the kidney-associated microbiome of a bat species abundant in Grenada, West Indies, Artibeus spp. Results indicate that the kidney of these bats can carry potential human pathogens. An endogenous retrovirus, Desmodus rotundus endogenous retrovirus isolate 824, phylogenetically related to betaretroviruses from rodents and New World primates, was also identified.

Tebuconazole-induced toxicity and the protective effect of Ficus carica extract in Neotropical fruit-eating bats.

Tebuconazole (TEB) is a triazole fungicide widely used in agriculture known to cause metabolic and endocrine disorders in mammals. Several plant extracts have shown to be beneficial against pesticide effects due to their hepatoprotective, antioxidant and anti-inflammatory properties. As fruit bats play a critical role in rainforest regeneration and are constantly exposed to pesticides, we aimed at evaluating TEB-induced toxicity and the possible protective effect of the Ficus carica plant extract in Neotropical fruit-eating bats (Artibeus lituratus). Bats were captured and assigned to 4 experimental groups, offered: 1) CTL (n = 6): papaya; 2) DMSO (n = 6): papaya treated with 1.25% dimethyl sulfoxide (DMSO); 3) TEB (n = 6): papaya treated with tebuconazole (commercial formulation) 0.1%; and 4) TEBFC (n = 6): papaya treated with tebuconazole 0.1% and Ficus carica extract (20%) in DMSO (1.25%). After seven days of exposure, TEB bats showed increased lipid peroxidation, increased superoxide dismutase (SOD) and catalase (CAT) activities, vascular congestion and inflammatory infiltrate in the liver, and increased serum transaminase enzyme activities. We found the same alterations in oxidative stress parameters in the breast muscles of TEB-exposed bats. In the testes, all oxidative stress markers were increased in TEB bats and corroborate findings of histopathological and increased serum testosterone levels observed following TEB exposure. The co-administration of the fungicide with the F. carica plant extract attenuated most oxidative stress markers in exposed bats' liver and testes and decreased liver damage, but failed to revert the steroid imbalance caused by the fungicide exposure.

Aspects regarding renal morphophysiology of fruit-eating and vampire bats.

Bats have adapted to many different feeding habits, which are known to induce morphophysiological adaptations in several tissues, especially those particularly involved with absorption, metabolism and excretion. The common vampire bat (Desmodus rotundus) has a very unique diet (blood), which, among other challenges, seems to pose a risk to their kidneys, due to the increased nitrogen excretion imposed by their remarkably high protein meal. Fruit-eating bats (Artibeus lituratus) consume a high carbohydrate diet and may be taken as a suitable species for this dietary comparative study. Here we aimed at investigating the renal morphology and stereology, kidneys antioxidant capacity, and plasma antidiuretic hormone (ADH) concentrations in adult fruit-eating and vampire bats. Sixteen animals were captured and used in this study, being 8 adult males from each species. Our results showed higher morphological standards of glomerular area, volumetric density of glomeruli, and renal somatic index for vampire bats, as well as higher reactive species of oxygen (ROS) production, such as nitric oxide (NO), higher plasma iron reduction ability (FRAP), higher activity of the antioxidant enzyme glutathione-S-transferase (GST) and a higher malondialdehyde production (MDA) in vampires' kidneys, compared to the fruit-eating species. The activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) were higher in fruit-eating bats. Plasma ADH concentrations were not different between species. Taken together, the renal morphophysiology conditions presented by vampire bats might be associated with a high demand for nitrogenous products excretion imposed by protein and iron overload. These features may play an important role on preventing protein-overload nephropathy, allowing vampires to survive under such a unique diet.

Trypanosoma Found in Synanthropic Mammals from Urban Forests of Paraná, Southern Brazil.

Trypanosoma cruzi is a parasitic protozoan that infects a diversity of hosts constituting the cycle of enzootic transmission in wild environments and causing disease in humans (Chagas disease) and domestic animals. Wild mammals constitute natural reservoirs of this parasite, which is transmitted by hematophagous kissing bugs of the family Reduviidae. T. cruzi is genetically subdivided into six discrete typing units (DTUs), T. cruzi (Tc)I to TcVI. In Brazil, especially in the state of Paraná, TcI and TcII are widely distributed. However, TcII is less frequently found in wild reservoirs and triatomine, and more frequently found in patients. The goal of this study was to investigate the natural occurrence of T. cruzi in wild synanthropic mammals captured in urban forest fragments of the Atlantic Forest of Paraná, southern Brazil. In this way, 12 opossums and 35 bats belonging to five species were captured in urban forest parks of the city of Maringá, Paraná, an area considered endemic for Chagas disease. PCR-kinetoplast DNA molecular diagnostic reveals Trypanosoma sp. infection in 12 (100%) Didelphis albiventris and 10 (40%) Artibeus lituratus. In addition to demonstrating the presence of Trypanosoma in the two groups of mammals studied, we obtained an isolate of the parasite genotyped as TcII by amplification of the cytochrome oxidase II gene by PCR, followed by restriction fragment length polymorphism with AluI, and confirmed by PCR of rDNA 24Sα. This is the first record of the encounter in wild mammals of Trypanosoma DNA (in A. lituratus) and T. cruzi DTU TcII (in D. albiventris) in the state of Paraná.

Natural exposure of bats in Grenada to rabies virus.

Introduction: Grenada is a rabies endemic country, where terrestrial rabies is maintained in the small Indian mongoose (Herpestes auropunctatus). The role of bats in the epidemiology of rabies in Grenada is unknown. A 1974 report described one rabies virus positive Jamaican fruit bat (Artibeus jamaicensis), and a high seroprevalence in this species. In the current study, the natural exposure to rabies virus in Grenadian bats was re-evaluated. It is postulated that bats serve as a natural rabies reservoir, probably circulating a bat-specific rabies virus variant. Material and methods: Bats were trapped in 2015 in all six parishes of Grenada using mist- and hand nets. For the detection of rabies virus in brain tissue, the direct fluorescent antibody test (dFAT) and the reverse transcription polymerase chain reaction (RT-PCR) were used. Serum neutralizing antibodies were determined using the fluorescent antibody virus neutralization test (FAVN). Results and discussion: Brain tissue and sera from 111 insectivorous and frugivorous bats belonging to four species were tested (52 Artibeus jamaicensis, two Artibeus lituratus, 33 Glossophaga longirostris, 24 Molossus molossus). Rabies virus antigen and genomic RNA were not detected in brain tissues. Rabies virus neutralizing antibodies were detected in the sera of eight A. jamaicensis in four of the six parishes. Bats in Grenada continue to show natural exposure to rabies virus. As rabies virus was not isolated in this study, serology alone is not sufficient to determine the strain of rabies virus circulating in A. jamaicensis bats in Grenada. Conclusion: Artibeus jamaicensis appears to play a role as a reservoir bat species, which is of public health concern in Grenada. Dispersion of bats to neighboring islands is possible and serological bat surveys should be initiated in these neighboring states, especially in those areas that are free of rabies in terrestrial mammals.