The genome sequence of Tadarida brasiliensis I. Geoffroy Saint-Hilaire, 1824 [Molossidae; Tadarida].

We present a genome assembly from an individual male Tadarida brasiliensis (The Brazilian free-tailed bat; Chordata; Mammalia; Chiroptera; Molossidae). The genome sequence is 2.28 Gb in span. The majority of the assembly is scaffolded into 25 chromosomal pseudomolecules, with the X and Y sex chromosomes assembled.

A paired analysis of mercury among non-invasive tissues to inform bat conservation monitoring.

Contaminant exposure can harm wildlife. However, measuring contaminant exposure in wildlife can be challenging due to accessibility of species and/or sampling tissue matrices needed to answer research questions regarding exposure. For example, in bats and other taxa that roost, it may be best to collect pooled feces from colonies for minimal disturbance to species of conservation concern, but fecal contaminant concentrations do not provide contaminant bioaccumulation estimates. Thus, there is a need for quantifying relationships between sample matrices for measuring contaminant exposure to answer research questions pertaining to wildlife health and addressing conservation needs. Our goal was to determine relationships between fecal and fur total mercury (THg). To do so, we collected paired feces and fur from Mexican free-tailed bats (Tadarida brasiliensis) in summer 2023 in western Oklahoma at a maternity roost with no known Hg point source. We analyzed THg in each sample matrix for each individual (n = 48). We found no relationship between individual fecal and fur THg. However, when averaged, fur THg was 6.11 times greater than fecal THg. This factor can be used as a screening-level risk assessment of under-roost feces, which could then be followed by direct assessments of fur THg concentrations and health impacts. We encourage the use of this conversion factor across other insectivorous bat species and sites for estimating initial risks of contaminant exposure with minimal disturbance to species of conservation concern, when timely research for conservation actions are needed, and when a contaminant point source is not yet known.

Living with voracious roommates: Factors that explain isotopic niche variation in a mixed colony of insectivorous bats.

Theory predicts that in resource-limited environments, coexisting species may overlap their niche dimensions but must differ in at least one to avoid competitive exclusion. Specifically, it has been suggested that the coexistence of competing species within a guild, could be sustained with mechanisms of resource partitioning, such as segregation along a trophic dimension. Among the most gregarious mammals are bats, which present diversification in their diet based on habitat choice and body size. Despite differences that could explain specialization in prey selection, there are insufficient studies that explore food overlap in mixed bat colonies and the factors that determine the selection of prey, both at intra- and inter-specific levels. To fill this gap, we analyzed the isotope signal (δ13C and δ15N) in feces collected in a mixed colony of Tadarida brasiliensis and Myotis chiloensis. To understand how several factors could influence these isotopic signals, intrinsic explanatory variables were analyzed, including body mass, body length, age, and sex. Also, extrinsic variables were analyzed, including monthly temporality and moonlight intensity. Our findings support age-dependent specialization in M. chiloensis, with a significant role of moonlight intensity and sex on δ15N. In T. brasiliensis, we identified a significant effect of size, sex, and ear length on δ15N. Our analysis indicates that both species of bats experience diverse degrees of overlap through austral summer months, affected by several factors that explain the variability in their fecal isotopic signals.

Geographically widespread and novel hemotropic mycoplasmas and bartonellae in Mexican free-tailed bats and sympatric North American bat species.

Bacterial pathogens remain poorly characterized in bats, especially in North America. We describe novel (and in some cases panmictic) hemoplasmas (12.9% positivity) and bartonellae (16.7% positivity) across three colonies of Mexican free-tailed bats (Tadarida brasiliensis), a partially migratory species that can seasonally travel hundreds of kilometers. Molecular analyses identified three novel Candidatus hemoplasma species most similar to another novel Candidatus species in Neotropical molossid bats. We also detected novel hemoplasmas in sympatric cave myotis (Myotis velifer) and pallid bats (Antrozous pallidus), with sequences in the latter 96.5% related to C. Mycoplasma haemohominis. We identified eight Bartonella genotypes, including those in cave myotis, with 96.7% similarity to C. Bartonella mayotimonensis. We also detected Bartonella rochalimae in migratory Tadarida brasiliensis, representing the first report of this human pathogen in bats. The seasonality and diversity of these bacteria observed here suggest that additional longitudinal, genomic, and immunological studies in bats are warranted.

Understanding fatality patterns and sex ratios of Brazilian free-tailed bats (Tadarida brasiliensis) at wind energy facilities in western California and Texas.

Background: Operation of wind turbines has resulted in collision fatalities for several bat species, and one proven method to reduce these fatalities is to limit wind turbine blade rotation (i.e., curtail turbines) when fatalities are expected to be highest. Implementation of curtailment can potentially be optimized by targeting times when females are most at risk, as the proportion of females limits the growth and stability of many bat populations. The Brazilian free-tailed bat (Tadarida brasiliensis) is the most common bat fatality at wind energy facilities in California and Texas, and yet there are few available data on the sex ratios of the carcasses that are found. Understanding the sex ratios of fatalities in California and Texas could aid in planning population conservation strategies such as informed curtailment. Methods: We used PCR to determine the sex of bat carcasses collected from wind energy facilities during post-construction monitoring (PCM) studies in California and Texas. In California, we received samples from two locations within the Altamont Pass Wind Resource Area in Alameda County: Golden Hills (GH) (n = 212) and Golden Hills North (GHN) (n = 312). In Texas, we received samples from three wind energy facilities: Los Mirasoles (LM) (Hidalgo County and Starr County) (n = 252), Los Vientos (LV) (Starr County) (n = 568), and Wind Farm A (WFA) (San Patricio County and Bee County) (n = 393). Results: In California, the sex ratios of fatalities did not differ from 50:50, and the sex ratio remained stable over the survey years, but the seasonal timing of peak fatalities was inconsistent. In 2017 and 2018, fatalities peaked between September and October, whereas in 2019 and 2020 fatalities peaked between May and June. In Texas, sex ratios of fatalities varied between locations, with Los Vientos being female-skewed and Wind Farm A being male-skewed. The sex ratio of fatalities was also inconsistent over time. Lastly, for each location in Texas with multiple years studied, we observed a decrease in the proportion of female fatalities over time. Discussion: We observed unexpected variation in the seasonal timing of peak fatalities in California and differences in the sex ratio of fatalities across time and facility location in Texas. In Texas, proximity to different roost types (bridge or cave) likely influenced the sex ratio of fatalities at wind energy facilities. Due to the inconsistencies in the timing of peak female fatalities, we were unable to determine an optimum curtailment period; however, there may be location-specific trends that warrant future investigation. More research should be done over the entirety of the bat active season to better understand these trends in Texas. In addition, standardization of PCM studies could assist future research efforts, enhance current monitoring efforts, and facilitate research on post-construction monitoring studies.

Detection of antimicrobial-resistant Enterobacterales in insectivorous bats from Chile.

Enterobacterales of clinical importance for humans and domestic animals are now commonly detected among wildlife worldwide. However, few studies have investigated their prevalence among bats, particularly in bat species living near humans. In this study, we assessed the occurrence of Extended-spectrum beta-lactamase-producing (ESBL) and carbapenemase-resistant (CR) Enterobacterales in rectal swabs of bats submitted to the Chilean national rabies surveillance program from 2021 to 2022. From the 307 swabs screened, 47 (15%) harboured cefotaxime-resistant Enterobacterales. Bats carrying these bacteria originated from 9 out of the 14 Chilean regions. Most positive samples were obtained from Tadarida brasiliensis (n = 42), but also Lasiurus varius, L. cinereus and Histiotus macrotus. No Enterobacterales were resistant to imipenem. All ESBL-Enterobacterales were confirmed as Rahnella aquatilis by MALDI-TOF. No other ESBL or CR Enterobacterales were detected. To our knowledge, this is the first screening of antibiotic-resistant bacteria in wild bats of Chile, showing the bat faecal carriage of R. aquatilis naturally resistant to cephalosporins, but also including acquired resistance to important antibiotics for public health such as amoxicillin with clavulanic acid. Our results suggest unknown selective pressures on R. aquatilis, but low or no carriage of ESBL or CR Escherichia coli and Klebsiella spp. Future studies should assess the zoonotic and environmental implications of R. aquatilis, which are likely present in the guano left by bats roosting in human infrastructures.

Diverse DNA virus genomes identified in fecal samples of Mexican free-tailed bats (Tadarida brasiliensis) captured in Chiricahua Mountains of southeast Arizona (USA).

Bats (order Chiroptera) are some of the most abundant mammals on earth and their species ecology strongly influences zoonotic potential. While substantial research has been conducted on bat-associated viruses, particularly on those that can cause disease in humans and/or livestock, globally, limited research has focused on endemic bats in the USA. The southwest region of the US is of particular interest because of its high diversity of bat species. We identified 39 single-stranded DNA virus genomes in the feces of Mexican free-tailed bats (Tadarida brasiliensis) sampled in the Rucker Canyon (Chiricahua Mountains) of southeast Arizona (USA). Twenty-eight of these belong to the virus families Circoviridae (n = 6), Genomoviridae (n = 17), and Microviridae (n = 5). Eleven viruses cluster with other unclassified cressdnaviruses. Most of the viruses identified represent new species. Further research on identification of novel bat-associated cressdnaviruses and microviruses is needed to provide greater insights regarding their co-evolution and ecology relative to bats.

Functional characterization of bat IRF1 in IFN induction.

Bats are natural hosts for various zoonotic viral diseases. However, they rarely show signs of disease infection with such viruses. During viral infection, members of the IRFs family induce the production of IFNß and exert antiviral effects. However, the functions of bat interferon regulatory factors (IRFs) remain unclear. In this study, the Tadarida brasiliensis IRF1 (TbIRF1) gene was first cloned and a series of bioinformatics studies were conducted. Results showed that bat IRF1 protein sequence showed a low similarity with IRF1s from other species. RNA virus such as Newcastle disease virus (NDV-GFP), avian influenza virus (AIV) and vesicular stomatitis virus (VSV-GFP) infection of Tadarida brasiliensis 1 lung (TB 1 Lu) cells significantly promotes the expressions of IFNß, PKR, and OAS1, and up-regulates the expression of TbIRF1. Overexpression of TbIRF1 markedly activates IFNß promoter activity in a dose-dependent manner. Next, we constructed the TbIRF1 functional domain deletion plasmids and found that the DNA binding domain (DBD) is necessary for TbIRF1 to induce IFNß expresison. In conclusion, the first bat IRF1 gene was cloned, and its functions in IFN induction were preliminarily identified.

Experimental Infection of Brazilian Free-Tailed Bats (Tadarida brasiliensis) with Two Strains of SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is presumed to have originated from wildlife and shares homology with other bat coronaviruses. Determining the susceptibility of North American bat species to SARS-CoV-2 is of utmost importance for making decisions regarding wildlife management, public health, and conservation. In this study, Brazilian free-tailed bats (Tadarida brasiliensis) were experimentally infected with two strains of SARS-CoV-2 (parental WA01 and Delta variant), evaluated for clinical disease, sampled for viral shedding and antibody production, and analyzed for pathology. None of the bats (n = 18) developed clinical disease associated with infection, shed infectious virus, or developed histopathological lesions associated with SARS-CoV-2 infection. All bats had low levels of viral RNA in oral swabs, six bats had low levels of viral RNA present in the lungs during acute infection, and one of the four bats that were maintained until 28 days post-infection developed a neutralizing antibody response. These findings suggest that Brazilian free-tailed bats are permissive to infection by SARS-CoV-2, but they are unlikely to contribute to environmental maintenance or transmission.

Occurrence of persistent organic pollutants and chlorpyrifos in Tadarida brasiliensis tissues from an agricultural production area in Argentina.

Organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) are lipophilic and persistent compounds, with chronic toxicity that cause negative effects on the environment and organisms. The chlorpyrifos is an organophosphate insecticide and represent the main insecticide currently used in many countries. The concentrations of OCPs, PCBs, and chlorpyrifos were evaluated in liver, muscle, and gonad of females and males of Tadarida brasiliensis in an agricultural production area in Argentina. Pollutants were quantified by gas chromatography with electron capture detection. The general total concentration pattern among tissues was gonad > muscle > liver, with a higher ratio of pesticides (OCPs + chlorpyrifos) over PCBs in muscle and liver, while gonads showed higher concentrations of PCBs. The predominance of DDTs and endosulfans evidences the bioavailability of these compounds despite their use has ceased. Chlorpyrifos was the main pesticide detected in almost all samples of both sexes, probably due to its current intensive use. More researches about ecology and activity patterns is paramount in order to understand more accurately the accumulation and possible negative effects of these pollutants.

Aerial attack strategies of hawks hunting bats, and the adaptive benefits of swarming.

Aggregation can reduce an individual's predation risk, by decreasing predator hunting efficiency or displacing predation onto others. Here, we explore how the behaviors of predator and prey influence catch success and predation risk in Swainson's hawks Buteo swainsoni attacking swarming Brazilian free-tailed bats Tadarida brasiliensis on emergence. Lone bats including stragglers have a high relative risk of predation, representing ~5% of the catch but ~0.2% of the population. Attacks on the column were no less successful than attacks on lone bats, so hunting efficiency is not decreased by group vigilance or confusion. Instead, lone bats were attacked disproportionately often, representing ~10% of all attacks. Swarming therefore displaces the burden of predation onto bats outside the column-whether as isolated wanderers not benefitting from dilution through attack abatement, or as peripheral stragglers suffering marginal predation and possible selfish herd effects. In contrast, the hawks' catch success depended only on the attack maneuvers that they employed, with the odds of success being more than trebled in attacks involving a high-speed stoop or rolling grab. Most attacks involved one of these two maneuvers, which therefore represent alternative rather than complementary tactics. Hence, whereas a bat's survival depends on maintaining column formation, a hawk's success does not depend on attacking lone bats-even though their tendency to do so is sufficient to explain the adaptive benefits of their prey's aggregation behavior. A hawk's success instead depends on the flight maneuvers it deploys, including the high-speed stoop that is characteristic of many raptors. Swarming bats emerging from a massive desert roost reduce their predation risk by maintaining tight column formation, because the hawks that predate them attack peripheral stragglers and isolated wanderers disproportionately. Whereas a bat's predation risk depends on maintaining its position within the column, the catch success of a hawk depends on how it maneuvers itself to attack, and is maximized by executing a high-speed dive or rolling grab maneuver.

Climatic drivers of (changes in) bat migration phenology at Bracken Cave (USA).

Climate change is drastically changing the timing of biological events across the globe. Changes in the phenology of seasonal migrations between the breeding and wintering grounds have been observed across biological taxa, including birds, mammals, and insects. For birds, strong links have been shown between changes in migration phenology and changes in weather conditions at the wintering, stopover, and breeding areas. For other animal taxa, the current understanding of, and evidence for, climate (change) influences on migration still remains rather limited, mainly due to the lack of long-term phenology datasets. Bracken Cave in Texas (USA) holds one of the largest bat colonies of the world. Using weather radar data, a unique 23-year (1995-2017) long time series was recently produced of the spring and autumn migration phenology of Brazilian free-tailed bats (Tadarida brasiliensis) at Bracken Cave. Here, we analyse these migration phenology time series in combination with gridded temperature, precipitation, and wind data across Mexico and southern USA, to identify the climatic drivers of (changes in) bat migration phenology. Perhaps surprisingly, our extensive spatiotemporal search did not find temperature to influence either spring or autumn migration. Instead, spring migration phenology seems to be predominantly driven by wind conditions at likely wintering or spring stopover areas during the migration period. Autumn migration phenology, on the other hand, seems to be dominated by precipitation to the east and north-east of Bracken Cave. Long-term changes towards more frequent migration and favourable wind conditions have, furthermore, allowed spring migration to occur 16 days earlier. Our results illustrate how some of the remaining knowledge gaps on the influence of climate (change) on bat migration and abundance can be addressed using weather radar analyses.

A Preliminary Study of the Virome of the South American Free-Tailed Bats (Tadarida brasiliensis) and Identification of Two Novel Mammalian Viruses.

Bats provide important ecosystem services as pollinators, seed dispersers, and/or insect controllers, but they have also been found harboring different viruses with zoonotic potential. Virome studies in bats distributed in Asia, Africa, Europe, and North America have increased dramatically over the past decade, whereas information on viruses infecting South American species is scarce. We explored the virome of Tadarida brasiliensis, an insectivorous New World bat species inhabiting a maternity colony in Rosario (Argentina), by a metagenomic approach. The analysis of five pooled oral/anal swab samples indicated the presence of 43 different taxonomic viral families infecting a wide range of hosts. By conventional nucleic acid detection techniques and/or bioinformatics approaches, the genomes of two novel viruses were completely covered clustering into the Papillomaviridae (Tadarida brasiliensis papillomavirus type 1, TbraPV1) and Genomoviridae (Tadarida brasiliensis gemykibivirus 1, TbGkyV1) families. TbraPV1 is the first papillomavirus type identified in this host and the prototype of a novel genus. TbGkyV1 is the first genomovirus reported in New World bats and constitutes a new species within the genus Gemykibivirus. Our findings extend the knowledge about oral/anal viromes of a South American bat species and contribute to understand the evolution and genetic diversity of the novel characterized viruses.

Experimental Infection of Tadarida brasiliensis with Pseudogymnoascus destructans, the Fungus That Causes White-Nose Syndrome.

White-nose syndrome (WNS) is causing significant declines in populations of North American hibernating bats, and recent western and southern expansions of the disease have placed additional species at risk. Understanding differences in species susceptibility and identifying management actions to reduce mortality of bats from WNS are top research priorities. However, the use of wild-caught susceptible bats, such as Myotis lucifugus, as model species for WNS research is problematic and places additional pressure on remnant populations. We investigated the feasibility of using Tadarida brasiliensis, a highly abundant species of bat that tolerates captivity, as the basis for an experimental animal model for WNS. Using methods previously established to confirm the etiology of WNS in M. lucifugus, we experimentally infected 11 T. brasiliensis bats with Pseudogymnoascus destructans in the laboratory under conditions that induced hibernation. We detected P. destructans on all 11 experimentally infected bats, 7 of which exhibited localized proliferation of hyphae within the epidermis, dermis, and subcutaneous tissue, similar to invasive cutaneous ascomycosis observed in M. lucifugus bats with WNS. However, the distribution of lesions across wing membranes of T. brasiliensis bats was limited, and only one discrete "cupping erosion," diagnostic for WNS, was identified. Thus, the rarity of lesions definitive for WNS suggests that T. brasiliensis does not likely represent an appropriate model for studying the pathophysiology of this disease. Nonetheless, the results of this study prompt questions concerning the potential for free-ranging, migratory T. brasiliensis bats to become infected with P. destructans and move the fungal pathogen between roost sites used by species susceptible to WNS.IMPORTANCE White-nose syndrome (WNS) is a fungal disease that is causing severe declines of bat populations in North America. Identifying ways to reduce the impacts of this disease is a priority but is inhibited by the lack of an experimental animal model that does not require the use of wild-caught bat species already impacted by WNS. We tested whether Tadarida brasiliensis, one of the most abundant species of bats in the Americas, could serve as a suitable animal model for WNS research. While T. brasiliensis bats were susceptible to experimental infection with the fungus under conditions that induced hibernation, the species exhibited limited pathology diagnostic for WNS. These results indicate that T. brasiliensis is not likely a suitable experimental model for WNS research. However, the recovery of viable WNS-causing fungus from experimentally infected bats indicates a potential for this species to contribute to the spread of the pathogen where it coexists with other species of bats affected by WNS.

Active acoustic interference elicits echolocation changes in heterospecific bats.

Echolocating bats often forage in the presence of both conspecific and heterospecific individuals, which have the potential to produce acoustic interference. Recent studies have shown that at least one bat species, the Brazilian free-tailed bat (Tadarida brasiliensis), produces specialized social signals that disrupt the sonar of conspecific competitors. We herein discuss the differences between passive and active jamming signals and test whether heterospecific jamming occurs in species overlapping spatiotemporally, as well as whether such interference elicits a jamming avoidance response. We compare the capture rates of tethered moths and the echolocation parameters of big brown bats (Eptesicus fuscus) challenged with the playback of the jamming signal normally produced by Brazilian free-tailed bats and playback of deconstructed versions of this signal. There were no differences in the capture rates of targets with and without the jamming signal, although significant changes in both spectral and temporal features of the bats' echolocation were observed. These changes are consistent with improvements of the signal-to-noise ratio in the presence of acoustic interference. Accordingly, we propose to expand the traditional definition of the jamming avoidance response, stating that echolocation changes in response to interference should decrease similarity between the two signals, to include any change that increases the ability to separate returning echoes from active jamming stimuli originating from conspecific and heterospecific organisms. Flexibility in echolocation is an important characteristic for overcoming various forms of acoustic interference and may serve a purpose in interspecific interactions as well as intraspecific ones.

Variation in Bat Guano Bacterial Community Composition With Depth.

Bats are known to be reservoirs for a variety of mammalian pathogens, including viruses, fungi, and bacteria. Many of the studies examining the microbial community inhabiting bats have investigated bacterial taxa found within specific bat tissues and isolated bat guano pellets, but relatively few studies have explored bacterial diversity within bat guano piles. In large bat caves, bat guano can accumulate over time, creating piles several meters deep and forming complex interactions with coprophagous organisms in a habitat with low light and oxygen. As the guano decays, the nutrient composition changes, but the bacterial communities deep within the pile have not been characterized. Here, we assess the bacterial communities across varying depths within the guano pile using both culture-independent and culture-dependent methods. We found that although similar taxa are found throughout the guano pile, the relative abundances of taxa within the pile shift, allowing certain taxa to dominate the bacterial community at varying depths. We also identified potential bacterial functions being performed within the bat guano as various depths within the pile and found little variation in terms of the dominant predicted functions, suggesting that although the relative abundances of bacterial taxa are changing, the functions being performed are similar. Additionally, we cultured 15 different bacterial species, including 2 not present in our culture-independent analysis, and discuss the pathogenicity potential of these taxa. This study represents the first characterization of the bacterial community from the extreme environment within a bat guano pile and demonstrates the potential for bat caves as resources for identifying new bacterial species.

Morphometric differences in testicular tissue of tadarida brasiliensis bats from the urban area of Mexico City during summer, autumn, and winter / Diferencias morfométricas en parenquima testicular de murcielagos tadarida brasiliensis del área urbana de la Ciudad de México en verano, otoño e invierno

Most of the neotropical bats reproduce in a seasonal fashion. The purpose of this study was to assess changes in testicular tissue by morphometry, in Tadarida brasiliensis bat, in a south urban zone of Mexico City during summer, autumn, and winter. Three sample collections (February, June, and September) from T. brasiliensis were carried out (n=6). Testicle fragments were obtained for histological studies. Diameter of the seminiferous tubules and interstitial space were measured, cellular populations were identified and counted. June samples showed smaller diameter of seminiferous tubules and larger interstitial space between tubules; also there were less number of germinal epithelium cells and spermatids were absent. Tissues from September and February showed a significant increase (p 0.05) in tubule diameter, germinal epithelium thickness, and number of germinal epithelium cells when compared to June samples. Only February samples showed presence of spermatids. Our results suggest the existence of seasonal variations in the reproductive activity of T. brasiliensis, under conditions in which the study was conducted.

La mayoría de los murciélagos neotropicales se reproducen de manera estacional. El objetivo de este estudio fue determinar los cambios morfométricos en el parénquima testicular del murciélago Tadarida brasiliensis que habita en un área urbana del sur de la Ciudad de México durante las estaciones de verano, otoño e invierno. Se obtuvieron fragmentos de tejido testicular para su estudio histológico. Se midió el diámetro de los túbulos seminíferos y el espacio interstisial y se identificaron y contaron distintos tipos celulares. En las muestras de junio se encontró un menor diámetro de los túbulos seminíferos y un mayor espacio intersticial entre los túbulos; también hubo un menor número de células del epitelio germinal y no hubo presencia de espermátides. En las muestras de tejido obtenidas en el mes de septiembre y febrero se observó un incremento significativo en el diámetro del túbulo, grosor del epitelio germinal y número de células del epitelio germinal cuando fueron comparadas con las muestras de junio (p 0.05). Solo en las muestras de febrero hubo presencia de espermátides. En conjunto, nuestros resultados sugieren la existencia de variaciones estacionales en la actividad reproductiva de T. brasiliensis, bajo las condiciones en que se realizó el estudio.

Dietary energetics of the insectivorous Mexican free-tailed bat (Tadarida brasiliensis) during pregnancy and lactation.

Stomach content analysis of 20 pregnant (x body mass=13.4 g) and 18 lactating (x body mass=11.5 g) female Tadarida brasiliensis revealed that the diet, expressed as percent volume, consists largely of lepidopterans, coleopterans, hymenopterans, and dipterans, in decreasing order of importance. We found no significant difference in the diet of pregnant and lactating females when expressed as percent volume. However, when expressed as percent frequency, proportionately more pregnant females fed on lepidopterans, coleopterans, and dipterans than did lactating bats, and proportionately more lactating females fed on hymenopterans. We found no significant differences in the percentages of water, lean dry mass, fat, and energy density in the stomach contents of pregnant and lactating females. Water in stomach contents averaged 62.7% and fat and lean dry mass averaged 22.2% and 15.2%, respectively (expressed as percentage of wet mass); energy density averaged 31.2 kJ g-1 dry mass. This relatively high energy density of stomach contents, as compared to whole insects, can be attributed to the consumption of insects high in fat (especially flying ants) and the abdomens only of moths and beetles (other body parts being discarded). Estimates of nightly food intake increased markedly from mid- to late pregnancy, stabilized or decreased during late pregnancy, and increased again during early to mid-lactation. Average nightly feeding rate doubled from pregnancy to lactation and increased threefold during the first half of lactation. By adjusting our gravimetric estimates of nightly feeding rate upwards by 40% (based on estimates of field metabolic rate), we estimate that the average nightly energy intake of female T. brasiliensis ranges from 57 kJ day-1 in early lactation to 104 kJ day-1 in mid-lactation. These estimates represent nightly feeding rates averaging 39% and 73% of a female's body mass in the period from early to mid-lactation, respectively.