Microbiota oral e retal de calitriquídeos (Callithrix sp) em área antropizada de Mata Atlântica, Rio de Janeiro, Brasil / [Oral and rectal microbiota of callitrichids (Callithrix sp) in an anthropized area of the Atlantic Forest in Rio de Janeiro, Brazil]

A proximidade dos primatas não humanos (PNH) com o ser humano pode ser considerada um fator de risco para transmissão de bactérias entre essas duas populações. Neste estudo, foi investigada a microbiota anfibiôntica aeróbica oral e retal de calitriquídeos em um fragmento de Mata Atlântica localizado no Rio de Janeiro, Brasil, e foram realizados testes fenotípicos para detecção de bactérias multirresistentes nos isolados encontrados. Foram capturados 14 calitriquídeos e coletadas 21 amostras (14 de cavidade oral e sete de cavidade retal) em dois pontos da mata próximos às habitações humanas. As espécies mais frequentes, na cavidade oral, foram Klebsiella oxytoca (50,0%), K. pneumoniae (28,6%), Kluyvera ascorbata (21,4%) e Stenotrophomonas maltophilia (21,4%) e, na cavidade retal, K. pneumoniae (85,7%), Escherichia coli (28,6%) e Enterobacter spp. (42,9%). Todos os 48 isolados da família Enterobacteriaceae foram negativos para ESBL (betalactamase de espectro ampliado), mostrando-se não produtores da enzima nos dois métodos utilizados: disco-aproximação e método de detecção automatizado. Na pesquisa de ERC (enterobactérias resistentes a carbapenêmicos), esses mesmos isolados não apresentaram resistência aos antibióticos imipenem, meropenem e ertapenem. Todas as bactérias isoladas apresentam um potencial zoonótico, o que representa um risco à saúde pública e à conservação das espécies.(AU)

Proximity of nonhuman primates (NHP) to humans can be considered a risk factor for transmission of pathogens between these two populations. This study investigated the oral and rectal aerobic bacterial microbiota of marmosets in an anthropized area of the Atlantic Forest located in Rio de Janeiro, Brazil, and performed phenotypic tests for detection of multidrug-resistant bacteria. Twenty-one samples (14 from the oral cavity and seven from the rectum) were collected from 14 Callithrix sp. captured in two sites of the forest near human dwellings. The most frequent species identified from the oral cavity swabs were Klebsiella oxytoca (50.0%), K. pneumoniae (28.6%), Kluyvera ascorbata (21.4%) and Stenotrophomonas maltophilia (21.4%), whereas the species most commonly identified from the rectum swabs were K. pneumoniae (85.7%), Enterobacter spp. (42.9%) and Escherichia coli (28.6%). All isolates of family Enterobacteriaceae showed no extended spectrum ß-lactamase production by disk-diffusion and automated detection tests. In the search for carbapenem-resistant enterobacteriaceae these isolates presented no resistance to the imipenem, meropenem and ertapenem antibiotics. The isolate of Staphylococcus aureus was susceptible to oxacillin and the isolate of Enterococcus was susceptible to vancomycin. All isolated bacteria showed zoonotic potential, thus posing a risk to species conservation and public health.(AU)

Involvement of midbrain tectum neurokinin-mediated mechanisms in fear and anxiety

Electrical stimulation of midbrain tectum structures, particularly the dorsal periaqueductal gray (dPAG) and inferior colliculus (IC), produces defensive responses, such as freezing and escape behavior. Freezing also ensues after termination of dPAG stimulation (post-stimulation freezing). These defensive reaction responses are critically mediated by γ-aminobutyric acid and 5-hydroxytryptamine mechanisms in the midbrain tectum. Neurokinins (NKs) also play a role in the mediation of dPAG stimulation-evoked fear, but how NK receptors are involved in the global processing and expression of fear at the level of the midbrain tectum is yet unclear. The present study investigated the role of NK-1 receptors in unconditioned defensive behavior induced by electrical stimulation of the dPAG and IC of male Wistar rats. Spantide (100 pmol/0.2 μL), a selective NK-1 antagonist, injected into these midbrain structures had anti-aversive effects on defensive responses and distress ultrasonic vocalizations induced by stimulation of the dPAG but not of the IC. Moreover, intra-dPAG injections of spantide did not influence post-stimulation freezing or alter exploratory behavior in rats subjected to the elevated plus maze. These results suggest that NK-1 receptors are mainly involved in the mediation of defensive behavior organized in the dPAG. Dorsal periaqueductal gray-evoked post-stimulation freezing was not affected by intra-dPAG injections of spantide, suggesting that NK-1-mediated mechanisms are only involved in the output mechanisms of defensive behavior and not involved in the processing of ascending aversive information from the dPAG.

Changes in the biogenic amine content of the prefrontal cortex, amygdala, dorsal hippocampus, and nucleus accumbens of rats submitted to single and repeated sessions of the elevated plus-maze test

It has been demonstrated that exposure to a variety of stressful experiences enhances fearful reactions when behavior is tested in current animal models of anxiety. Until now, no study has examined the neurochemical changes during the test and retest sessions of rats submitted to the elevated plus maze (EPM). The present study uses a new approach (HPLC) by looking at the changes in dopamine and serotonin levels in the prefrontal cortex, amygdala, dorsal hippocampus, and nucleus accumbens in animals upon single or double exposure to the EPM (one-trial tolerance). The study involved two experiments: i) saline or midazolam (0.5 mg/kg) before the first trial, and ii) saline or midazolam before the second trial. For the biochemical analysis a control group injected with saline and not tested in the EPM was included. Stressful stimuli in the EPM were able to elicit one-trial tolerance to midazolam on re-exposure (61.01 percent). Significant decreases in serotonin contents occurred in the prefrontal cortex (38.74 percent), amygdala (78.96 percent), dorsal hippocampus (70.33 percent), and nucleus accumbens (73.58 percent) of the animals tested in the EPM (P < 0.05 in all cases in relation to controls not exposed to the EPM). A significant decrease in dopamine content was also observed in the amygdala (54.74 percent, P < 0.05). These changes were maintained across trials. There was no change in the turnover rates of these monoamines. We suggest that exposure to the EPM causes reduced monoaminergic neurotransmission activity in limbic structures, which appears to underlie the "one-trial tolerance" phenomenon.

Routine post-weaning handling of rats prevents isolation rearing-induced deficit in prepulse inhibition

Rats reared under isolation conditions from weaning present a number of behavioral changes compared to animals reared under social conditions (group housing). These changes include deficits in prepulse inhibition (PPI) of the startle reflex to a loud sound. PPI refers to the reduction of the magnitude of the startle reflex when a relatively weak stimulus (the prepulse) precedes by an appropriate time interval the intense startle-elicing stimulus (the pulse). PPI is useful for studying sensorimotor integration. The present study evaluated the effect of handling on the impairment of PPI induced by isolation-rearing. Male Wistar rats (N = 11-15/group) were housed in groups (5 per cage and handled three times a week) or isolated (housed individually) since weaning (21 days) for 10 weeks when they reach approximately 150 g. The isolated rats were divided into "minimally handled" animals (handled once a week for cleaning purposes only) or "handled" animals (handled three times a week). This handling consisted of grasping the rat by the tail and moving it to a clean cage (approximately 5 s). A statistically significant reduction (52 percent) in the PPI test was found only in the isolated group with minimal handling while no difference was seen between grouped animals and isolated handled animals. These results indicate that isolation rearing causes disruption in the PPI at adult age, which serves as an index of attention deficit. This change in the sensory processing of information induced by post-weaning isolation can be prevented by handling during the development of the animal.

Anatomical connections of the periaqueductal gray: specific neural substrates for different kinds of fear

The periaqueductal gray (PAG) has been traditionally considered to be an exit relay for defensive responses. Functional mapping of its subdivisions has advanced our knowledge of this structure, but synthesis remains difficult mainly because results from lesion and stimulation studies have not correlated perfectly. After using a strategy that combined both techniques and a reevaluation of the available literature on PAG function and connections, we propose here that freezing could be mediated by different PAG subdivisions depending on the presence of immediate danger or exposure to related signaling cues. These subdivisions are separate functional entities with distinct descending and ascending connections that are likely to play a role in different defensive responses. The existence of ascending connections also suggests that the PAG is not simply a final common path for defensive responses. For example, the possibility that indirect ascending connections to the cingulate cortex could play a role in the expression of freezing evoked by activation of the neural substrate of fear in the dorsal PAG has been considered

The brain decade in debate: II. Panic or anxiety? From animal models to a neurobiological basis

This article is a transcription of an electronic symposium sponsored by the Brazilian Society of Neuroscience and Behavior (SBNeC). Invited researchers from the European Union, North America and Brazil discussed two issues on anxiety, namely whether panic is a very intense anxiety or something else, and what aspects of clinical anxiety are reproduced by animal models. Concerning the first issue, most participants agreed that generalized anxiety and panic disorder are different on the basis of clinical manifestations, drug response and animal models. Also, underlying brain structures, neurotransmitter modulation and hormonal changes seem to involve important differences. It is also common knowledge that existing animal models generate different types of fear/anxiety. A challenge for future research is to establish a good correlation between animal models and nosological classification

Signaled two-way avoidance learning using eletrical stimulation of the inferior colliculus as negative reinforcement: effects of visual and auditory cues as warning stimuli

The inferior colliculus is a primary relay for the processing of auditory information in the brainstem. The inferior colluculus is also part of the so-called brain aversion system as animals learn to switch off the electrical stimulation of this structure. The purpose of the present study was to determine whether associative learning occurs between aversion induced by electrical stimulation of the inferior colliculus and visual and auditory warning stimuli. Rats implanted with electrodes into the central nucleus of the inferior colliculus were placed inside an open-field and thresholds for the escape response to electrical stimulation of the inferior colliculus were determined. The rats were then placed inside a shuttle-box and submitted to a two-way avoidance pardigm. Electrical stimulation of the inferior colliculus at the escape threshold (98.12 + 6.15 (A, peak-to-peak) was used as negative reinforcement and light or tone as the warning stimulus. Each session consisted of 50 trials and was divided into two segments of 25 trials in order to determine the learning rate of the animals during the sessions. The rats learned to avoid the inferior colliculus stimulation when light was used as the warning stimulus (13.25 + 0.60 s and 8.63 + 0.93 for lactencies and 12.5 + 2.04 and 19.62 + 1.65 frequencies in the first and second halves of the sessions, respectively, P<0.01 in both cases). No significant changes in latencies (14.75 + 1.63 and 12.75 + 1.44 s) or frequencies of responses (8.75 + 1.20 and 11.25 + 1.13) were seen when tone was used as the warning stimulus (P>0.05 in both cases). Taken together, the present results suggest that rats learn to avoid the inferior colliculus stimulation when light is used as the warning stimulus. However, this learning process does not occur when the neutral stimulus used is an acoustic one. Electrical stimulation of the inferior colliculus may disturb the signal transmission of the stimulus to be conditioned from the inferior colliculus to higher brain structures such as amygdala.

Paradoxical increase of exploratory behavior in the elevated plus-maze by rats exposed to two kinds of aversive stimuli

Albino rats were submitted to a 24-h period of social isolation (individual housing) combined with 0, 1, 2 or 3 twenty-four-hour periods of exposure to different vivaria (novelty) and tested in the elevated plus-maze. Results, reported as mean + SEM for N = 12, show that the time (in seconds) spent in the open arms by rats exposed to novelty for 0, 1,2 and 3 days was 28.3 + 4.4, 31.6 + 3.2, 29.1 + 3.5 and 25.0 + 3.3, respectively, when grouped in the same vivarium; 29.6 + 2.7, 7.6 + 2.1, 9.6 + 4.4 and 28.5 + 3.7 when grouped in different vivaria; 2.9 + 1.1, 1.8 + 1.0, 2.7 + 1.1 and 0 + 0 when isolated in the same vivarium, and 2.6 + 1.1, 31.5 + 8.2, 24.8 + 4.2 and 0 + 0 when isolated in different vivaria. The number of entries into the open and closed arms followed a similar trend. This indicates that, separately, both exposure to novelty and isolation are aversive manipulations. Paradoxically, when novelty was combined with a concomitant 24-h period of social isolation prior to testing, the decrease in exploratory behavior caused by either of the two aversive manipulations alone was reverted. These results are indicative that less intense anxiety triggers mechanisms mediating less energetic behavior such as freezing, while higher levels trigger mechanisms mediating more vigorous action, such as flight/fight behavior, since the combination of two aversive situations resulted in more exploratory behavior than with either alone. They are also suggestive of habituation to the effects of novelty, since exposure to it for 3 days produced exploratory behavior similar to that of controls.

Behavioral and pharmacological validation of the elevated plus maze contructed with transparent walls

In this study we compared the performance of male Wistar rats, weighing 250-300g, submitted to the standard plus maze (vertical surfaces of the closed arms with opaque walls) to their performance in a modified maze with raised Plexiglas edges in the closed arms (transparent walls). The animals (N = 12 for each group) continued to show a clear preference for the closed arms with transparent walls of the modified elevated plus maze. In addition, exploratory activity was higher in the open arms of the modified plus maze (4.25 ñ 0.42 entries and 53.50 ñ 5.10s) as compared to that of the standard plus maze (2.10 ñ 0.25 entries and 24.00 ñ 4.91 s). Intraperitoneal injection of midazolam produced an increase in the number of entries (6.40 ñ 1.21 and 8.50 ñ 1.15 for 1.0 and 2.0 mg/Kg, rspectively) and in the time spent in the open arms (85.32 ñ 14.56 and 125.50 ñ 22.16 s for 1.0 and 2.0 mg/Kg, respectively) while pentylenetetrazole caused a decrease in the number of entries (3.68 ñ 0.54 and 2.33 ñ 0.62 for 5.0 and 10 mg/Kg, respectively) and in the time spent in the open arms of the modified maze (39.60 ñ 6.67 and 23.60 ñ 6.40 s for 5.0 and 10 mg/Kg, respectively). The anxiolytic effect of midazolam and the anxiogenic effect of pentylenetetrazole were similar to those usually reported in the literature by authors using the standard test. The4se results behaviorally and pharmacologically validate the elevated plus maze with transparebnt walls and suggest that this test could be a useful tool for the study of anxiolytic drugs and the neurobiology of anxiety

Behavioral effects of neurotensin applied to periventricular structures of rats

Neurotensin (NT), n active neuropeptide, and bicuculline, a GABA-A receptor antagonist, were microinjected into the rat hypothalaamus (MH) or the dorsal periaqueductal gray matter (DPAG). Bicuculline (80 pmol) produced behavioral activation which included jumping and NT (1-20 nmol) caused a dose-dependent behavioral activation accompanied by catalepsy rather than jumping. These results suggest that the behavioral activation produced by NT may be due to an interaction of the neuropeptide with specific receptors while its cataleptic effect may be attributed to the blockade of dopamine receptors

Increased susceptibility of detelencephalated rats to audiogenic seizures induced by microinjection of bicuculline into the inferior colliculus

The participation of telencephalic forebrain structures in the induction of audiogenic seizure (AGS) susceptibility and in the behavioral expression of AGS was investigated in rats. Rats that were initially susceptible (N = 12) or non-susceptible (N = 28) to audiogenic seizure were surgically detelencephalated. A unilateral microinjection of a low dose (30 pmol) of the GABA antagonist bicuculline methiodide (BM) was applied to the inferior colliculus (IC) before the animals were exposed to a 120-dB acoustic stimulus. All susceptible rats still exhibited all components of audiogenic seizure after removal of the telencephalon. After BM microinjection, a higher incidence (66% vs 41%) and shorter latencies (6-20 s vs 9-55) s) of occurrence of tonic seizures were observed in the detelencephalated non-susceptible rats when compared to non-operated non-susceptible rats(N = 12). These results suggest that the induction of the behavioral expression of audiogenic seizures issubserved by brain stem neuronal networks but does not require the telencephalon and that telencephalic structures may exert control over audiogenic seizures by inhibiting IC cells through GABAergic neurons

Gabaergic fibers from substantia nigra para reticulata modulate escape bahavior induced by midbrain central gray stimulation

Eletrical stimulation or microinjection of GABA antagonists into the dorsal periaqueductal gray (DPAG) produces escape behavior. In order to determine whether the nigrocollicular gabaergic fibers exert some control over this behavior, rats bearing kainic acid lesion of the substantia nigra pars reticulata were submitted to microinjections of bicuculline or electrical stimulation of the DPAG at the escape threshold. Rats thus treated exhibited a significant decrease in the escape threshold while bicuculline increased the expression of flight behavior. These results suggest an inhibitory control of gabaergic fibers from the substantia nigra pars reticulata on aversive behavior induced by DPAG stimulation

Independence of aversive and pain mechanisms in the dorsal periaqueductal gray matter of the rat

Electrical stimulation of the dorsal periaqueductal gray (DPAG) elicits autonomic responses similar to those following peripheral pain stimulation. The present study analyzes the effects of morphine and midazolan applied to the same DPAG sites on the autonomic responses induced by both types of stimulation. Both drugs attenuated the increase in heart rate and blood pressure induced by DPAG stimulation while attenuating only the increase in heart rat induced by pain stimulation. These results suggest that the neural substrates of the autonomic expression of the DPAG and pain stimulation are different although they may partially overlap