Serological evidence of hantavirus infection in neotropical bats in an urban area of São Paulo State, Brazil.

BACKGROUND: Although hantaviruses have long been associated with rodents, they are also described in other mammalian hosts, such as shrews, moles and bats. Hantaviruses associated with bats have been described in Asian, European and Brazilian species of bats. As these mammals represent the second major mammalian order, and they are the major mammals that inhabit urban areas, it is extremely important to maintain a viral surveillance in these animals. Our aim was to conduct serosurveillance in bats in an urban area in the city of Ribeirão Preto, São Paulo State, Brazil, to contribute to the information about hantaviruses circulation in bats. METHODS: We analyzed samples from 778 neotropical bat specimens classified into 21 bat species and four different families collected in the urban area of Ribeirão Preto city, from 2014 to 2019 by an ELISA for the detection of IgG antibodies against orthohantavirus. RESULTS: We detected IgG-specific antibodies against the nucleoprotein of orthohantavirus in 0.9% (7/778) bats tested, including four Molossus molossus (Pallas' Free-tailed Bat), two Glossophaga soricina (Pallas's Long-tongued Bat) and one Eumops glaucinus (Wagner's mastiff bat). CONCLUSIONS: Overall, our results show the first serological evidence of hantavirus infection in three common bat species in urban areas.

Quantification of Urinary Sex Steroids in the Big Brown Bat (Eptesicus fuscus).

AbstractBats (order Chiroptera) are the second largest group of mammals, diverging ~52.5 million years ago. Many species exhibit an unusual reproductive cycle and extreme longevity without reproductive senescence, yet steroid profiles exist for few bats. Big brown bats (Eptesicus fuscus) are temperate insectivores found throughout North America. They mate promiscuously in fall, store sperm during winter hibernation, and have delayed ovulation and fertilization in spring. Here, we report the first urinary steroid profile in bats by quantifying 17ß-estradiol (E2) in captive male and female E. fuscus across their reproductive cycle. Male bats had higher urinary E2 levels than females, and adults had higher levels than yearlings following creatinine adjustment for hydration. In nonpregnant females, several seasonal differences in creatinine-adjusted and unadjusted urinary E2 levels were observed. Urinary E2 was higher in males than females in winter for both conditions and in autumn for creatinine-adjusted levels. We quantified progesterone (P4) in a subset of females. In nonpregnant females, urinary P4 was constant across seasons except for unadjusted levels, which were highest in the summer. In pregnant females, urinary E2 and P4 levels peaked beginning ~20 d before parturition, with both steroids returning to baseline in the following weeks. Knowing how urinary steroid levels fluctuate with age and sex and across the annual season is key to understanding reproductive cycling in bats. Our research furthers the potential for bats as a model for medical reproductive research. Moreover, it complements previous studies on the potential role of steroids in primer pheromonal effects in bats.

An Outbreak in Pigeons Caused by the Subgenotype VI.2.1.2 of Newcastle Disease Virus in Brazil.

Newcastle disease virus (NDV) can infect over 250 bird species with variable pathogenicity; it can also infect humans in rare cases. The present study investigated an outbreak in feral pigeons in São Paulo city, Brazil, in 2019. Affected birds displayed neurological signs, and hemorrhages were observed in different tissues. Histopathology changes with infiltration of mononuclear inflammatory cells were also found in the brain, kidney, proventriculus, heart, and spleen. NDV staining was detected by immunohistochemistry. Twenty-seven out of thirty-four tested samples (swabs and tissues) were positive for Newcastle disease virus by RT-qPCR test, targeting the M gene. One isolate, obtained from a pool of positive swab samples, was characterized by the intracerebral pathogenicity index (ICPI) and the hemagglutination inhibition (HI) tests. This isolate had an ICPI of 0.99, confirming a virulent NDV strain. The monoclonal antibody 617/161, which recognizes a distinct epitope in pigeon NDV strains, inhibited the isolate with an HI titer of 512. A complete genome of NDV was obtained using next-generation sequencing. Phylogenetic analysis based on the complete CDS F gene grouped the detected isolate with other viruses from subgenotype VI.2.1.2, class II, including one previously reported in Southern Brazil in 2014. This study reports a comprehensive characterization of the subgenotype VI.2.1.2, which seems to have been circulating in Brazilian urban areas since 2014. Due to the zoonotic risk of NDV, virus surveillance in feral pigeons should also be systematically performed in urban areas.

Histomorphology of the glans penis in Vespertilionidae and Phyllostomidae species (Chiroptera, Mammalia).

The penises of bats are taxonomically distinctive in size and shape. In addition, they are variable in microscopic anatomy, indicating that histomorphological studies of copulatory organs of bats may help understanding their successful reproductive strategies. We studied adult males of 13 species of vespertilionid and phyllostomid bats. Both families exhibited the basic structure of the vascular penis of mammals: the hydrostatic elements of the corpora cavernosa and the corpus spongiosum surrounding the urethra, as well as accessory cavernous tissue. Variation in the position and amount of the tissues were observed in these families. Vespertilionid bats have a small glans penis with abundant accessory cavernous tissue on the prepuce and a highly variable baculum. The baculum varied in size and morphology, even among congeneric species, such as the three Lasiurus species and the two Myotis species. Phyllostomid species possess no bacula, but vascular structures are present to produce penile stiffening, particularly on the glans. Variation in the microscopic anatomy of the phyllostomid prepuce was observed, for example, Artibeus species had accessory cavernous tissue surrounded by a tunica albuginea, but Carollia perspicillata had two bundles of striated musculature and some adipose tissue; abundant pigments were present in the prepuce of most species.

Lack of serological and molecular evidence of arbovirus infections in bats from Brazil.

Viruses are important agents of emerging zoonoses and are a substantial public health issue. Among emerging viruses, an important group are arboviruses, which are characterized by being maintained in nature in cycles involving hematophagous arthropod vectors and a wide range of vertebrate hosts. Recently, bats have received increasing attention as an important source for the emergence of zoonoses and as possible viral reservoirs. Among the arboviruses, there are many representatives of the genera Flavivirus and Alphavirus, which are responsible for important epidemics such as Dengue virus, Zika virus and Chikungunya virus. Due to the importance of analyzing potential viral reservoirs for zoonosis control and expanding our knowledge of bat viruses, this study aimed to investigate the presence of viruses of the Alphavirus and Flavivirus genera in bats. We analyzed serum, liver, lungs and intestine from 103 bats sampled in northeast and southern Brazil via Nested-PCR and the hemagglutination inhibition test. All samples tested in this study were negative for arboviruses, suggesting that no active or past infection was present in the captured bats. These data indicate that the bats examined herein probably do not constitute a reservoir for these viruses in the studied areas. Further studies are needed to clarify the role of bats as reservoirs and sources of infection of these viral zoonoses.

Morphological and histological characters of penile organization in eleven species of molossid bats.

The penis is the reproductive organ that ensures efficient copulation and success of internal fertilization in all species of mammals, with special challenges for bats, where copulation can occur during flight. Comparative anatomical analyses of different species of bats can contribute to a better understanding of morphological diversity of this organ, concerning organization and function. In this study, we describe the external morphology and histomorphology of the penis and baculum in eleven species of molossid bats. The present study showed that penile organization in these species displayed the basic vascular mammalian pattern and had a similar pattern concerning the presence of the tissues constituting the penis, exhibiting three types of erectile tissue (the corpus cavernosum, accessory cavernous tissue, and corpus spongiosum) around the urethra. However, certain features varied among the species, demonstrating that most species are distinguishable by glans and baculum morphology and glans histological organization. Major variations in glans morphology were genus-specific, and the greatest similarities were shared by Eumops species and N. laticaudatus. The greatest interspecific similarities occurred between M. molossus and M. rufus and between Eumops species. Save for M. molossus and M. rufus, morphology of the baculum was species-specific; and in E. perotis, it did not occur in all specimens, indicating that it is probably under selection. In the histological organization, the most evident differences were number of septa and localization of the corpora cavernosa. In species with a baculum (Molossus, Eumops and Nyctinomops species), the corpora cavernosa predominantly occupied the dorsal region of the penile glans and is associated with the proximal (basal) portion of the baculum. In species that do not have a baculum (Cynomops, Molossops and Neoplatymops species), the corpora cavernosa predominantly occupied the ventro-lateral region of the glans.

Structure, histochemistry, ultrastructure and seasonal variations of the male prostatic complex in the black Myotis bat, Myotis nigricans (Chiroptera: Vespertilionidae).

Chiroptera are one of the most diverse orders of mammals and a unique group within Mammalia that posses a wide geographic distribution and considerable variability in reproductive strategies. The aims of the present study were to characterise the male prostatic complex of the bat Myotis nigricans (Vespertilionidae) and evaluate seasonal variations in the prostatic complex of M. nigricans specifically. Twenty-three sexually mature specimens (four sample groups: winter, spring, summer and autumn) were subjected to macroscopic, microscopic, morphometric and ultrastructural analyses. The reproductive accessory glands of M. nigricans were found to be composed of a multilobed complex associated with the urethra and a pair of inguinal bulbourethral glands. The complex was composed of three bilobed prostatic regions (ventral, dorsolateral and dorsal) with no ampullary gland and seminal vesicles. This pattern of lobulation is very similar to that described for the prostate of rodents; however, it differs from that of other mammals and even other families of bats (e.g. Phyllostomidae and Molossidae). Each prostatic region in M. nigricans has unique and distinctive characteristics, which synchronise to establish the main reproductive peak of the species in summer. The data also indicated an asynchrony in the activity of primary and secondary reproductive organs in the annual reproductive cycle of M. nigricans in São Paulo State, Brazil.

Ultrastructure of spermatogenesis, spermatozoon and processes of testicular regression and recrudescence in Eptesicus furinalis (Chiroptera: Vespertilionidae).

Studies have shown that the annual reproductive cycle of Eptesicus furinalis includes at least one period of total testicular regression. Thus, this study aimed to evaluate their reproductive cycle ultrastructurally. The annual reproductive cycle was divided into four periods: active, regressing, regressed and recrudescence. The active period was similar to that of other bats, including the completion of spermatogenesis with three main types of spermatogonia (Ad, Ap and B) and 12 steps in the process of spermatid differentiation. However, its spermatozoa differed in that outer dense fibers 1, 5, 6 and 9 are larger than the others and due to the presence of what is likely a probably genera-specific bulging in the anterior portion. In the regressing period, Sertoli cell nuclei migrate to the basal compartment with the nuclei close to the basal lamina. The basal compartment had a more compact appearance than the adluminal compartment, with relaxed cellular connections. In the regressed period, spermatogenesis ceased; the seminiferous epithelium was composed only of Sertoli cells and three types of spermatogonia: types Ad, 1 and 2. In the recrudescence period, spermatogenesis restarted, with the process of reactivation divided into three phases: early, medial and late recrudescence. In conclusion, our study described the process of spermatogenesis and the ultrastructure of the spermatozoa and confirmed the presence of a process of total testicular regression in the annual cycle of E. furinalis. We characterize distinct morphologic variations in the ultrastructure of the testicular cells during the four different periods of the annual reproductive cycle.

Ultrastructure of spermatogenesis in the short-tailed fruit bat, Carollia perspicillata (Chiroptera: Phyllostomidae: Carollinae).

Among species of the Chiroptera, spermatogenesis and the fully differentiated spermatozoa differ in morphological and ultrastructural detail. This study therefore aimed to ultrastructurally characterize the spermatogenesis and the spermatozoa of Carollia perspicillata (Phyllostomidae) and compare the process with other species of bats and mammals. The differentiation of spermatogonia is similar to other bats and to Primates, with three main spermatogonia types: Ad, Ap, and B. Meiotic divisions proceed similarly to those of most mammals and spermiogenesis is clearly divided into 12 steps, in the middle of the range of developmental steps for bats (9-16 steps). The process of acrosome formation is similar to that found in Platyrrhinus lineatus, with the acrosome formed by two different types of proacrosomal vesicles. The ultrastructure of the spermatozoon is similar to other bats already described and resembles the typical mammalian sperm model; however, its morphology differs from other mammals such as marsupials and rodents, on account of a simpler spermatozoon head morphology, which indicates a pattern that is more closely related to the sperm cells of humans and other primates. Our data demonstrated that spermatogenesis in C. perspicillata presents great ultrastructural similarities to P. lineatus. This pattern is not surprising, because both species belong to the same family (Phyllostomidae); however, it is observed that C. perspicillata presents some characteristics that are more closely related to phylogenetically distant species, such as Myotis nigricans (Vespertilionidae), which is a fact that deserves attention.