Sex hormones change visceral pigmentation in Eupemphix nattereri (Anura): effects in testicular melanocytes and hepatic melanomacrophages

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Valores hematológicos de morcegos hematófagos Desmodus rotundus (E. Geoffroy, 1810) mantidos em cativeiro

Hematological values from captive vampire bats Desmodus rotundus (E. Geoffroy, 1810). Desmodus rotundus is one of the hematophagous bat species that are responsible for significant losses to livestock and has important involvement on public health. The great interest on this bat species made it becomes the target of many investigations a required its maintenance in laboratories. Similarly to others mammals, hematological evaluation has been utilized to assess the health and morbidity state of bats, however there are scarce studies with captive bats. The aim of this study was to investigate the hypothesis that is possible to feed the vampire bat D. rotundus with frozen blood treated with citrate during a long captivity period without hematological dyscrasias. Therefore, complete blood count was performed monthly from 15 adult females kept for 345 days in cages and fed with frozen blood added to citrate. The erythrocyte concentration (9.02 ± 1.43 x1012/L), PCV (0.47 ± 0.08 L/L) and hemoglobin (163.9 ± 31.5 g/L) obtained from free-living bats (immediately after capture) were lower or similar to those obtained after 345 days of captivity, presenting erythrocytes’ count of 11.01 ± 0.82 x1012/L, packed cell volume of 0.50 ± 0.05 L/L and hemoglobin level of 158 ± 10.1 g/L. The total white blood cell (11.09 ± 6.07 x109 /L) and segmented neutrophil counts (9.85 ± 3.5 x109 /L) of free living D. rotundus decreased significantly after 345 days of captivity, with values of 3.98 ± 1.98 and 1.87 ± 978.6 x109 /L respectively, which are similar to bats from temperate regions in hibernation period. This study proved that is possible to feed D. rotundus for long periods of captivity with citrated blood without the occurrence of anemia, erythrocyte or other hematologic dyscrasia

Seasonal Carbon Dioxide and Oxygen Concentrations in the Dens of Hibernating Mammals (Sciuridae)

This paper reports seasonal changes in respiratory gases in artificial dens of two species of hibernators indigenous to central and northern Alaska: Citellus parryi ablusus(Osgood) and Marmota broweri Hall and Gilmore (Sciuridae).

Räumliche und zeitliche Einbindung einer strukturierten Population der Westlichen Smaragdeidechse (Lacerta bilineata, DAUDIN 1802) im Mittelrheintal

In der vorliegenden Arbeit wurde die räumliche und zeitliche Dynamik einer strukturierten Population der Westlichen Smaragdeidechse (DAUDIN, 1802) im Mittelrheintal zwischen 1997 und 2000 untersucht. Dazu wurden die Teilaspekte der Hibernation, der Thermoregulation und der Raumnutzung der Art analysiert. Die Ergebnisse der Hibernation deuten auf eine hohe Relevanz dieses Jahresabschnitts für die Art in Bezug auf die Verbreitung in Südexponierten Steilhanglagen am Nordrand ihrer Verbreitung hin. Die Hibernationsstandorte unterscheiden sich im Mikroklimaprofil von Trockenmauern und Sommerlöchern. Die Standorte der Hibernation liegen in Zonen höchster Insolation im Untersuchungsgebiet. Die Analyse der Thermoregulation zeigte geschlechtsspezifische und saisonale Unterschiede, die mit den unterschiedlichen Jahresaktivitätszyklen erklärt werden können. Die Weibchen zeigen eine engere Bindung an die klimatischen Entwicklungen von Mirkohabitatstrukturen als die Männchen, was mit endogenen Faktoren zusammenhängen könnte. Die Parallelen und Unterschiede zu anderen Arten werden diskutiert. In der Raumnutzung zeigen sich auch Unterschiede zwischen Männchen und Weibchen, die auch mit den unterschiedlichen Jahresaktivitätszyklen erklärt werden können. Im Frühjahr und im Herbst zeigt die Art eine Bindung an Standorte hoher Insolation im Untersuchungsgebiet, im Sommer dagegen keine. In der Gesamtbetrachtung werden die Ergebnisse in Bezug auf das Verbreitungsbild der Art, Schutzaspekte und Schlüsselfaktoren der Habitatqualität diskutiert.

Aspetti ipnici e vegetativi dell'inibizione del controllo nervoso centrale della termogenesi nel ratto

La possibilità di indurre stati ipotermici ed ipometabolici come il torpore o l’ibernazione in animali non ibernanti può avere dei risvolti utili nella pratica medica, in quanto permetterebbe di trarre vantaggio dagli effetti benefici dell’ipotermia senza gli effetti compensatori negativi causati dalla risposta omeostatica dell’organismo. Con questo lavoro vogliamo proporre un nuovo approccio, che coinvolge il blocco farmacologico dell’attività dei neuroni nel bulbo rostroventromediale (RVMM), un nucleo troncoencefalico che si è rivelato essere uno snodo chiave nella regolazione della termogenesi attraverso il controllo dell’attività del tessuto adiposo bruno, della vasomozione cutanea e del cuore. Nel nostro esperimento, sei iniezioni consecutive del agonista GABAA muscimolo nel RVMM, inducono uno stato reversibile di profonda ipotermia (21°C al Nadir) in ratti esposti ad una temperatura ambientale di 15°C. Lo stato ipotermico/ipomentabolico prodotto dall’inibizione dei neuroni del RVMM mostra forti similitudini col torpore naturale, anche per quanto concerne le modificazioni elettroencefalografiche osservate durante e dopo la procedura. Come negli ibernati naturali, nei ratti cui viene inibito il controllo della termogenesi si osserva uno spostamento verso le regioni lente delle spettro di tutte le frequenze dello spettro EEG durante l’ipotermia, ed un forte incremento dello spettro EEG dopo il ritorno alla normotermia, in particolare della banda Delta (0,5-4Hz) durante il sonno NREM. Per concludere, questi risultati dimostrano che l’inibizione farmacologica selettiva di un nucleo troncoencefalico chiave nel controllo della termogenesi è sufficiente per indurre uno stato di psuedo-torpore nel ratto, una specie che non presenta stati di torpore spontaneo. Un approccio di questo tipo può aprire nuove prospettive per l’utilizzo in ambito medico dell’ipotermia.

Electrolyte Depletion in White-nose Syndrome Bats

The emerging wildlife disease white-nose syndrome is causing widespread mortality in hibernating North American bats. White-nose syndrome occurs when the fungus Geomyces destructans infects the living skin of bats during hibernation, but links between infection and mortality are underexplored. We analyzed blood from hibernating bats and compared blood electrolyte levels to wing damage caused by the fungus. Sodium and chloride tended to decrease as wing damage increased in severity. Depletion of these electrolytes suggests that infected bats may become hypotonically dehydrated during winter. Although bats regularly arouse from hibernation to drink during winter, water available in hibernacula may not contain sufficient electrolytes to offset winter losses caused by disease. Damage to bat wings from G. destructans may cause life-threatening electrolyte imbalances.

Altered Behavior in Bats Affected by White-Nose Syndrome

WNS-affected bats did so over similar time frames as WNSunaffected bats. The behaviors of bats with WNS did not change as drastically as expected. Thereseems to be little to no effect on their ability to fly/forage until much later stages of the disease when they are likely near death. WNS-affected bats are grooming more which could be altering the way they use energy reserves during hibernation possibly leading tostarvation and eventually death. The decreased likelihood of arousals in response to external cues may be the result of spending more energy during previous and increasingly frequent arousals. While it is clear that WNS does result in changes in behavior whether these changes are directly in response to fungal skin infection or to some other component of the syndrome such as decreased energy availability or loss of homeostasis is unknown. bat behavior, white-nose syndrome, behavior, video surveillance, arousal patterns White-Nose Syndrome (WNS) is a disease of hibernating bats caused by the fungal pathogen Geomyces destructans. The fungus, which was first noted in 2006, invades bats wings and other exposed membranes, eventually resulting in death. Researchers have yet to understand many aspects of this disease, including basic etiology and epidemiology. There is also a lack of information on how fungal infection may change the behavior of healthy bats during hibernation or how changes in behavior may influence disease progression. Based upon the physiological changes that are known to occur in affected bats, and upon anecdotal observations of aberrant behavior in these bats, I hypothesized that WNS would significantly change the behavior of the little brown myotis (Myotis lucifugus). My research examined the behavior of hibernating bats during arousals from torpor. I compared WNS-affected and unaffected bats, in the field and incaptivity, using motion-sensitive infrared cameras. Flight maneuverability and echolocation were also tested between WNS-affected and unaffected bats during arousalsfrom hibernation to detect changes in the bats' ability to perform basic locomotion or potentially catch insect prey. Lastly, hibernating bats were artificially disturbed and theirarousal patterns were monitored to examine changes in the response to external stimuli between WNS-affected and unaffected bats.Bats with WNS groomed for longer periods of time after arousing from torpor, both in the field and in captivity. They also engaged in longer periods of any sort of activity during these arousals. There were no changes in acoustical signaling during flight tests and changes in flight maneuverability were only found in bats were seen staging" near the entrance of the mine which is itself a unique behavior exhibited by affected bats. At this point these bats were likely near death and could barely fly at all. In response toexternal stimuli bats with WNS were less likely to arouse than unaffected bats. However when they did arouse WNS-affected bats did so over similar time frames as WNSunaffected bats. The behaviors of bats with WNS did not change as drastically as expected. Thereseems to be little to no effect on their ability to fly/forage until much later stages of the disease when they are likely near death. WNS-affected bats are grooming more which could be altering the way they use energy reserves during hibernation possibly leading tostarvation and eventually death. The decreased likelihood of arousals in response to external cues may be the result of spending more energy during previous and increasingly frequent arousals. While it is clear that WNS does result in changes in behavior whetherthese changes are directly in response to fungal skin infection or to some other component of the syndrome such as decreased energy availability or loss of homeostasis is unknown."

Physiological Consequences Of White-Nose Syndrome

White-nose syndrome (WNS) is a disease that has caused the mass mortality of hibernating bat species. Since its first discovery in the winter of 2006-2007, an estimated five million bats or more have been killed. Although infection with Pseudogymnoascus destructans (Pd, the causative agent of WNS) does not always result in death, bats that survive Pd infection may experience fitness consequences. To understand the physiological consequences of WNS, I measured reproductive rates of free-ranging hibernating bat species of the Northeastern United States. In addition, captive little brown myotis (Myotis lucifugus) bats that were infected by Pd but survived (¿WNS survivors¿) and uninfected bats were studied in order to understand the potential consequences (e.g., lower reproductive rates, decreased ability to heal wounds, degradation of wing tissue, and altered metabolic rates) of surviving WNS. No differences in reproductive rates were found between WNS-survivors and uninfected bats in either the field or in captivity. In addition, wound healing was not affected by Pd infection. However, wing tissue degradation was worse for little brown myotis 19 days post-hibernation, and mass specific metabolic rate (MSMR) was significantly higher for those infected with Pd 22 days post-hibernation. While it is clear that these consequences are a direct result of Pd infection, further research investigating the long-term consequences for both mothers and pups is necessary.

Exploration Of Survival Traits In The White-Nose Syndrome-Affected Big Brown Bat (Eptesicus Fuscus)

The widespread mortality of hibernating bats is associated with the emerging infectious disease white-nose syndrome (WNS), and has provoked a strong interest in understanding which bats will survive, and why? The ability of infected bats to resist WNS may depend upon variation in the expression of different characteristics. In a captive colony of big brown bats, I sought to characterize the phenotypic variability, repeatability, and survivability for several key ¿survival¿ traits, including: torpor patterns, microclimate preferences, and wound healing capacity. Torpor patterns were profiled using temperature sensitive dataloggers throughout the hibernation season, while microclimate preferences were quantified by using temperature-graded boxes and thermal imaging. In order to assess wound healing capacity, small wing biopsies were obtained from each bat and healing progress was tracked for one month. Individuals exhibited a wide range of phenotypes that were significantly influenced by sex and body condition. Repeatability estimates suggest that there is not a strong genetic basis for the observed variation in torpor patterns or microclimate preferences. Certain phenotypes (e.g., BMI) were associated with an increased probability of overwinter survivorship, which suggests a basis for intra-species differences in WNS susceptibility. The results from this project provide novel insight into what we know about ¿who will survive,¿ and will influence the direction and implementation of future conservation and mitigation strategies.

Perinatal adaptation in mammals: the impact of metabolic rate

Mammalian birth is accompanied by profound changes in metabolic rate that can be described in terms of body size relationship (Kleiber's rule). Whereas the fetus, probably as an adaptation to the low intrauterine pO2, exhibits an "inappropriately" low, adult-like specific metabolic rate, the term neonate undergoes a rapid metabolic increase up to the level to be expected from body size. A similar, albeit slowed, "switching-on" of metabolic size allometry is found in human preterm neonates whereas animals that are normally born in a very immature state are able to retard or even suppress the postnatal metabolic increase in favor of weight gain and O2 supply. Moreover, small immature mammalian neonates exhibit a temporary oxyconforming behavior which enhances their hypoxia tolerance, yet is lost to the extent by which the size-adjusted metabolic rate is "locked" by increasing mitochondrial density. Beyond the perinatal period, there are no other deviations from metabolic size allometry among mammals except in hibernation where the temporary "switching-off" of Kleiber's rule is accompanied by a deep reduction in tissue pO2. This gives support to the hypothesis that the postnatal metabolic increase represents an "escape from oxygen" similar to the evolutionary roots of mitochondrial respiration, and that the overall increase in specific metabolic rate with decreasing size might contribute to prevent tissues from O2 toxicity.

Treatment of osteoporosis in a mouse model of Duchenne muscular dystrophy using black bear parathyroid hormone

Duchenne muscular dystrophy (DMD) is a progressive disease affecting skeletal and cardiac muscle, as well as bone. Long term disuse and glucocorticoid treatments cause progressive osteoporosis in DMD patients, leading to an increase in fracture incidence. Treatments for osteoporosis in these patients have not been widely explored. Parathyroid hormone (PTH), an anabolic treatment for post-menopausal osteoporosis, could benefit DMD patients by improving skeletal properties and reducing fracture risk. Other PTH analogues are not currently FDA approved to treat osteoporosis, but may have improved osteogenic effects compared to the human analogue. Black bear PTH is especially promising as an osteoporosis treatment for the DMD population. Black bears are unique models of bone maintenance during disuse, since during six months of inactivity (hibernation), they maintain skeletal properties, unlike other hibernators. Additionally, black bear PTH has been correlated to bone formation markers during hibernation, indicating it may be, at least in part, the mechanism by which bears maintain bone during disuse. Employing black bear PTH as a treatment for osteoporosis in DMD patients could greatly improve quality of life for these individuals, and reduce the pain and expense associated with frequent fractures.

Fatal combined infection with canine distemper virus and orthopoxvirus in a group of Asian marmots (Marmota caudata)

A fatal combined infection with canine distemper virus (CDV) and orthopoxvirus (OPXV) in Asian marmots (Marmota caudata) is reported in this article. A total of 7 Asian marmots from a small zoological garden in Switzerland were found dead in hibernation during a routine check in the winter of 2011. The marmots died in February 2011. No clinical signs of disease were observed at any time. The viruses were detected in all individuals for which the tissues were available (n = 3). Detection of the viruses was performed by reverse transcription polymerase chain reaction. The most consistent gross lesion was a neck and thorax edema. A necrotizing pharyngitis and a multifocal necrotizing pneumonia were observed histologically. Numerous large intracytoplasmic eosinophilic inclusions were seen in the epithelial cells of the pharynx, of the airways, and in the skin keratinocytes. Brain lesions were limited to mild multifocal gliosis. Phylogenetic analysis revealed that the marmot CDV strain was closely related to the clusters of CDVs detected in Switzerland in wild carnivores during a local outbreak in 2002 and the 2009-2010 nationwide epidemic, suggesting a spillover of this virus from wildlife. The OPXV was most closely related to a strain of cowpoxvirus, a poxvirus species considered endemic in Europe. This is the first reported instance of CDV infection in a rodent species and of a combined CDV and OPXV infection.

Detection of Chlamydia pneumoniae in a collection of captive snakes and response to treatment with marbofloxacin.

In a collection of 58 snakes comprising predominantly Eurasian vipers in Switzerland, five snakes died unexpectedly during hibernation from 2009 to 2012. In one snake, organisms resembling chlamydiae were detected by immunohistochemistry in multiple histiocytic granulomas. Real-time quantitative PCR and microarray analysis were used to determine the presence of Chlamydia pneumoniae in tissue samples and cloacal/choanal swabs from snakes in the collection; 8/53 (15.1%) of the remaining snakes were positive. Although one infected snake had suppurative periglossitis, infection with C. pneumoniae did not appear to be associated with specific clinical signs in snakes. Of seven snakes treated with 5 mg/kg marbofloxacin IM once daily, five became PCR negative for C. pneumoniae following treatment, whereas one animal remained positive and one snake was lost to follow-up.

Torpor in mice is induced by both leptin-dependent and -independent mechanisms

We tested the effect of chronic leptin treatment on fasting-induced torpor in leptin-deficient A-ZIP/F-1 and ob/ob mice. A-ZIP/F-1 mice have virtually no white adipose tissue and low leptin levels, whereas ob/ob mice have an abundance of fat but no leptin. These two models allowed us to examine the roles of adipose tissue and leptin in the regulation of entry into torpor. Torpor is a short-term hibernation-like state that allows conservation of metabolic fuels. We first characterized the A-ZIP/F-1 animals, which have a 10-fold reduction in total body triglyceride stores. Upon fasting, A-ZIP/F-1 mice develop a lower metabolic rate and decreased plasma glucose, insulin, and triglyceride levels, with no increase in free fatty acids or β-hydroxybutyrate. Unlike control mice, by 24 hr of fasting, they have nearly exhausted their triglycerides and are catabolizing protein. To conserve energy supplies during fasting, A-ZIP/F-1 (but not control) mice entered deep torpor, with a minimum core body temperature of 24°C, 2°C above ambient. In ob/ob mice, fasting-induced torpor was completely reversed by leptin treatment. In contrast, neither leptin nor thyroid hormone prevented torpor in A-ZIP/F-1 mice. These data suggest that there are at least two signals for entry into torpor in mice, a low leptin level and another signal that is independent of leptin and thyroid hormone levels. Studying rodent torpor provides insight into human torpor-like states such as near drowning in cold water and induced hypothermia for surgery.

Long-term suppression of Leydig cell steroidogenesis prevents Leydig cell aging

Male aging is accompanied by reduced testosterone production by the Leydig cells, the testosterone-producing cells of the testis. The mechanism by which this occurs is unknown. Based on the observations that reactive oxygen is capable of damaging components of the steroidogenic pathway and that reactive oxygen is produced during steroidogenesis itself, we hypothesized that long-term suppression of steroidogenesis might inhibit or prevent age-related deficits in Leydig cell testosterone production. To test this, we administered contraceptive doses of testosterone to groups of young (3 months old) and middle-aged (13 months old) Brown Norway rats via Silastic implants to suppress endogenous Leydig cell testosterone production. After 8 months, the implants were removed, which rapidly (days) restores the ability of the previously suppressed Leydig cells to produce testosterone. Two months after removing the implants, when the rats of the two groups were 13 and 23 months of age, respectively, the Leydig cells in both cases were found to produce testosterone at the high levels of young Leydig cells, whereas significantly lower levels were produced by the 23-month-old controls. Thus, by placing the Leydig cells in a state of steroidogenic “hibernation,” the reductions in Leydig cell testosterone production that invariably accompany aging did not occur. If hormonal contraception in the human functions the same way, the adverse consequences of reduced testosterone in later life (osteoporosis, reduced muscle mass, reduced libido, mood swings, etc.) might be delayed or prevented.