Strain by the train: Patterns of toad fatalities on a Brazilian Amazonian railroad.

Transportation infrastructures are directly responsible for killing billions of animals worldwide. Although the understanding about road impacts have recently increased, the impact of railroads on wildlife has received less attention. The current knowledge concerning the impacts of railroads focuses mainly on large mammals although amphibians might be affected. Our study aims to unravel temporal and spatial patterns of Rhinella toad fatalities on a Brazilian Amazonian railroad, to comprehend how toads are killed and to estimate the magnitude of toad fatalities. Data collection was carried out on foot on an 871-km stretch of the Estrada de Ferro Carajás from 2013 to 2017. We identified different potential causes for fatalities: being run over, desiccated or with barotrauma signs. We estimated a surprisingly high carcass persistence probability of about 38 days. After correcting for the bias from carcass detection and removal, we estimated that approximately 10,000 toads are killed per year (≈ 11 fatalities/km/year). A GLM model showed that toads were more likely to be killed in the dry to wet transition. We identified critical zones of fatalities and prioritized them according to their intensity. The highly critical segments encompass >10% of all fatalities although they cover only 1.5% of the railroad. Our study is the first one to address carcass detection and persistence on railroads and to unravel patterns of fatalities of an amphibian species in a tropical climate. A better understanding of the patterns of animal fatality on railroads is of fundamental importance to manage and mitigate this impact.

BRAZIL ROAD-KILL: a data set of wildlife terrestrial vertebrate road-kills.

Mortality from collision with vehicles is the most visible impact of road traffic on wildlife. Mortality due to roads (hereafter road-kill) can affect the dynamic of populations of many species and can, therefore, increase the risk of local decline or extinction. This is especially true in Brazil, where plans for road network upgrading and expansion overlaps biodiversity hotspot areas, which are of high importance for global conservation. Researchers, conservationists and road planners face the challenge to define a national strategy for road mitigation and wildlife conservation. The main goal of this dataset is a compilation of geo-referenced road-kill data from published and unpublished road surveys. This is the first Data Paper in the BRAZIL series (see ATLANTIC, NEOTROPICAL, and BRAZIL collections of Data Papers published in Ecology), which aims make public road-kill data for species in the Brazilian Regions. The dataset encompasses road-kill records from 45 personal communications and 26 studies published in peer-reviewed journals, theses and reports. The road-kill dataset comprises 21,512 records, 83% of which are identified to the species level (n = 450 species). The dataset includes records of 31 amphibian species, 90 reptile species, 229 bird species, and 99 mammal species. One species is classified as Endangered, eight as Vulnerable and twelve as Near Threatened. The species with the highest number of records are: Didelphis albiventris (n = 1,549), Volatinia jacarina (n = 1,238), Cerdocyon thous (n = 1,135), Helicops infrataeniatus (n = 802), and Rhinella icterica (n = 692). Most of the records came from southern Brazil. However, observations of the road-kill incidence for non-Least Concern species are more spread across the country. This dataset can be used to identify which taxa seems to be vulnerable to traffic, analyze temporal and spatial patterns of road-kill at local, regional and national scales and also used to understand the effects of road-kill on population persistence. It may also contribute to studies that aims to understand the influence of landscape and environmental influences on road-kills, improve our knowledge on road-related strategies on biodiversity conservation and be used as complementary information on large-scale and macroecological studies. No copyright or proprietary restrictions are associated with the use of this data set other than citation of this Data Paper.

Roosters affected by epididymal lithiasis present local alteration in vitamin D3, testosterone and estradiol levels as well as estrogen receptor 2 (beta) expression.

Epididymal lithiasis is a reproductive dysfunction of roosters that is associated with loss of fertility and is characterized by the formation of calcium stones in the lumen of the efferent ductules of the epididymal region. The efferent ductules of birds are responsible for the reabsorption of the fluid coming from the testis as well as luminal calcium. It has been hypothesized that the epididymal stone formation may be related to the impairment of local fluid or calcium homeostasis, which depends on hormones such as estradiol (E(2)). Therefore, this study aimed to investigate possible alterations in the expression of ERα (ESR1) and ERß (ESR2) in the epididymal region of roosters affected by epididymal lithiasis. The study was performed by immunohistochemistry and western blotting assays. In addition, the concentrations of E(2), vitamin D3, and testosterone, which are also key hormones in maintenance of calcium homeostasis, were determined in the plasma and epididymal region, by ELISA. It was observed that ESR2 expression is increased in all segments of the epididymal region of affected roosters, whereas ESR1 levels are not altered. Moreover, the hormone concentration profiles were changed, as in the epididymal region of roosters with lithiasis the E(2) levels were increased and vitamin D3 as well as testosterone concentrations were significantly decreased. These results suggest that a hormonal imbalance may be involved with the origin and progression of the epididymal lithiasis, possibly by affecting the local fluid or calcium homeostasis.

Occurrence and cellular distribution of estrogen receptors ERα and ERß in the testis and epididymal region of roosters.

Estrogen signaling is required for the maintenance of male reproductive function and is mediated by the estrogen receptors ERα and ERß. These receptors are widely distributed in mammalian reproductive tissues, but information is limited in non-mammalian species including birds. The aim of this study was to investigate the occurrence and cellular distribution of ERα and ERß in the testis and epididymal region of roosters. The results showed for the first time that ERß was the predominant receptor detected in the testis, being expressed in the somatic and some germ cells. Within the epididymal region, ERß was strongly expressed in all segments, whereas the most intense reaction for ERα was found in the distal efferent ductules. The differential expression of ERα and ERß within the rooster testis and epididymal region suggests that these organs may be a target for different actions of estrogen.

Vitamin D3 and androgen receptors in testis and epididymal region of roosters (Gallus domesticus) as affected by epididymal lithiasis.

Epididymal lithiasis is a dysfunction characterized by formation of calcium-rich stones in the epididymal region of roosters, associated with decreased serum testosterone and loss of fertility. The segment most affected by the lithiasis is the efferent ductules, which, in birds, are responsible for reabsorption of calcium and luminal fluid. Therefore, we postulated that epididymal lithiasis could result from local impairment of calcium or fluid homeostasis, culminating in initiation of stone formation. Transepithelial calcium transport depends on vitamin D3 and vitamin D3 receptor (VDR). Based on the fact that VDR are present in efferent ductules, possible changes in the pattern of VDR in roosters affected by the epididymal lithiasis was investigated, to start to gain an understanding of the molecular mechanisms involved in the development of calcium stones. To evaluate the potential impact of androgen reduction, changes in androgen receptor (AR) were also investigated. Both VDR and AR were increased in specific segments of the epididymal region, whereas no alterations were found in the testes of affected animals. The increase in VDR was most likely due to an increase in the number of VDR-positive mononuclear leukocyte infiltrates found in the connective tissue followed by an increase in epithelial receptors. The AR were increased, however, mainly in the epididymal duct epithelium. These results suggest that the vitamin D3 and androgen responsive system may be directly/indirectly involved in the development of the disease.

Comparative expression of androgen receptor in the testis and epididymal region of roosters (Gallus domesticus) and drakes (Anas platyrhynchos).

The androgen receptor (AR) mediates the physiological actions of androgens, which play a crucial role in the maintenance of male reproductive function and fertility. Although the AR distribution pattern is well established in mammalian reproductive organs, information about the AR expression in the testes and epididymal region of birds is still scarce. To better clarify the pattern of AR expression in the avian male tract, we investigated the expression and precise cellular distribution of AR in the testis and epididymal region of roosters and drakes. AR expression was investigated by immunohistochemistry and Western blotting. In the testis, AR was found restricted to the nuclei of Sertoli cells, Leydig cells and some myoid cells in both species. Within the epididymal region, AR was widely expressed in the epithelia of all segments, although with segment specific differences in intensity and cellular distribution. Stronger positivity for AR was found in the principal cells of the epididymal duct, followed by the rete testis epithelium and non-ciliated cells of the distal efferent ductules. Non-ciliated cells of the proximal efferent ductules epithelium showed the lowest immunostaining. Ciliated cells of both segments of the efferent ductules were negative for AR. The connective tissue of roosters presented fewer AR-positive cells when compared with drakes; despite the similar total number of cells in both species. In conclusion, cellular and segment specific differences in AR expression suggest difference in sensitivity to androgens among the ducts composing the epididymal region of roosters and drakes.

Distribution of vitamin D3 receptor in the epididymal region of roosters (Gallus domesticus) is cell and segment specific.

Vitamin D3 is a steroid hormone well known by its role in maintaining calcium homeostasis, however this hormone may also participate in other biological functions, including control of reproductive processes. The vitamin D3 action is mediated by the vitamin D3 receptor (VDR). VDR is widely distributed in the rodent reproductive tract, however the occurrence of VDR and the role of the vitamin D3 in the avian reproductive tract remain unknown. The aim of the present study was to investigate the expression and cellular distribution of VDR in the epididymal region of roosters. VDR expression was investigated by Western blotting analysis and the tissue distribution of the receptor was determined by immunohistochemistry. The Western blotting assay revealed a major VDR protein band of 61kDa in the epididymal region of rooster. Nuclear VDR expression was found in all segments of the epididymal region, namely rete testis, efferent ductules, connecting ducts and epididymal ducts. Nonciliated cells of the distal efferent ductules showed the highest levels of VDR expression, followed by the proximal efferent ductules and rete testis. The connecting and epididymal ducts showed less intense VDR immunostaining. The differential VDR expression in the epididymal region segments reveals that several extratesticular ducts may be target for vitamin D3 action and suggests that vitamin D3 may have a regional-specific function, such as calcium transport, that is modulated through VDR activity.