Role of cholinergic, opioidergic and GABAergic neurotransmission of the dorsal hippocampus in the modulation of nociception in guinea pigs

Aims: Several physiological, pharmacological and behavioral lines of evidence suggest that the hippocampal formation is involved in nociception. The hippocampus is also believed to play an important role in the affective and motivational components of pain perception. Thus, Our aim was to investigate the participation of cholinergic, opioidergic and GABAergic systems of the dorsal hippocampus (DH) in the modulation of nociception in guinea pigs. Main methods: The test used consisted of the application of a peripheral noxious stimulus (electric shock) that provokes the emission of a vocalization response by the animal. Key findings: Our results showed that, in guinea pigs, microinjection of carbachol, morphine and bicuculline into the DH Promoted anti nociception, while muscimol promoted pronociception. These results were verified by a decrease and all increase, respectively, in the vocalization index in the vocalization test. This antinociceptive effect of carbachol (2.7 nmol) was blocked by previous administration of atropine (0.7 nmol) or naloxone (1.3 nmol) into the same site. In addition, the decrease in the vocalization index induced by the microinjection of morphine (2.2 nmol) into the DH was prevented by pretreatment with naloxone (1.3 nmol) or muscimol (0.5 nmol). At doses of 1.0 nmol, muscimol microinjection caused pronociception, while bicuculline promoted antinociception. Significance: These results indicate the involvement of the cholinergic, opioidergic and GABAergic systems of the DH in the modulation of antinociception in guinea pigs. In addition, the present study suggests that cholinergic transmission may activate the release of endorphins/enkephalin from interneurons of the DH, Which Would inhibit GABAergic neurons, resulting in antinociception. (C) 2008 Elsevier Inc. All rights reserved.

Histological effects of intratympanic gentamicin on the vestibular organ of guinea pigs

Background: Transtympanic administration of gentamicin may be suitable to achieve unilateral vestibular ablation, in order to control unilateral Meniere`s disease. In low doses, gentamicin appears to affect selectively the vestibular system, with relative sparing of the cochlea. An experimental study on guinea pigs was conducted to determine what single dose of gentamicin would produce a unilateral vestibular organ lesion when applied to the middle ear. Study design: Experimental and prospective. Methods: Four groups of guinea pigs received different gentamicin doses ( 1, 5, 10 and 25 mg) administered to the middle ear. The animals` vestibular organs were then assessed by scanning electron microscopy, in order to quantify the level of vestibular damage. Results: Study of the utricular macula and the ampullar crista of the lateral semicircular canal revealed vestibular neuroepithelial lesions in all infused ears. Conclusions: The severity of the vestibular neuroepithelial lesions was dose-dependent. Lower gentamicin doses were observed to damage vestibular structures more than cochlear structures.

Organophosphate-related ototoxicity: Description of the vestibulocochlear system ultrastructural aspects of guinea pigs

Organophosphate toxic agents are used in agriculture and are currently part of the group of toxic agents which can lead to hearing loss, in which we have solvents, metals and asphyxiation agents. Aim: to analyze the acute ototoxic action of a group of organophosphate agents in the vestibulocochlear system. This is a prospective experimental study. Materials and Methods: we used male albino guinea pigs, broken down into three groups, to which we provided distilled water (group 1 - control), agrotoxic agent - 0.3mg/Kg/day (group 2), agrotoxic - 3 mg/Kg/day (group 3), during 7 seven consecutive days. The most used agrotoxic agent was Tamaron BR (metamidophos). The anatomical evaluation of the cochlea, saccule and utricle was carried out by means of electronic scanning microscopy after the use of the agrotoxic agent. Results: the guinea pigs submitted to the organophosphate presented cochlear morphological alterations with lesions on the three turns analyzed, as well as cilia alterations in the saccule and utricle, intensified according to the agent dosage. Conclusion: the morphological alterations seen in the hair cells exposed to daily doses of organophosphate promote evidences of an acute deleterious effect of agrotoxic agents on the vestibulocochlear system.

IRRIGATION OF UROPIGIAN GLAND OF DOMESTIC FOWLS (Gallus gallus domesticus) OF COBB LINEAGE

In this study was evaluated the vascular system of uropigian gland in 34 fowls (Gallus gallus domesticus), of Cobb lineage. The material was donated by the firm Globo Aves, located in Uberlandia - MG. Was specially studied the origin of collaterals that are aimed to that gland. Animals had them arterial contingents injected with a colored solution of neoprene latex, by polyethylene canulas through the right isquiatic artery. Then was injected an aqueous solution of formol 10%, in subcutaneous and intracavity spaces, which made possible the material fixation. Finally was proceeded the individually dissection of circumscribed region of uropigian gland. Opportunely was used a monocular lens in observation of vascular profile and was prepared schematics models representing the irrigation of each fowl. Uropigian gland showed itself as irrigated by mediana caudal artery in all (100%) animals, through its intersegmentaries branches, which emit right and left lateral glandular branchs and, right and left medials. Was counted the sub-branches, and was verified bigger emission in the right by the M. D. 1,1 and 1,2; and in the left by the M. E. 1,1 and 1,2.

Evaluation of IFA, MAT, ELISAs and immunoblotting for the detection of anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs

The study evaluated the efficiency of diagnostic laboratory methods to detect anti-Toxoplasma gondii antibodies in paired serum and aqueous humour samples from experimentally infected pigs. 18-mixed breed pigs were used during the experiment; these were divided into two groups, G I (infected group, n = 10) and G2 (uninfected group, n = 8). Infection was performed with 4 x 10(4) VEG strain oocysts at day 0 by the oral route in G1 animals. All pigs were euthanized at day 60, when retina, aqueous humour, and blood samples were collected. Anti-T gondii antibody levels were assessed in serum (s) and aqueous humour (ah) by indirect immunofluorescence assay (IFA), modified agglutination test (MAT), m-ELISA (using crude membranes from T gondii tachyzoites as antigen) and r-ELISA (using rhoptries from T gondii tachyzoites as antigen). Polymerase chain reactions (PCR) of samples from the retina were performed by using Tox4 and Tox5 primers. Antibody titers of G1 animals ranged from 128 to 1024 and from 16 to 256 in serum and aqueous humour, respectively. There were differences in the correlation coefficients between IFA(s) x IFA (ah) (r = 0.62, P = 0.05), MAT(s) x MAT (ah) (r = 0.97, P < 0.0001); however, there was no significant difference between r-ELISA(s) x r-ELISA (ah) (r = 0. 14, P = 0.7). Antibodies present in serum and aqueous humour recognized similar antigens. Samples of retina were positive by PCR in 30% (3/10) of infected pigs. G2 animals remained without antibody levels and were PCR negative throughout the experiment. These results suggest that the use of a combination of tests and immunoblotting for paired aqueous humour and serum samples could improve the sensitivity and specificity for the diagnosis of ocular toxoplasmosis. (c) 2007 Elsevier Ltd. All rights reserved.

Rickettsial infection in domestic mammals and their ectoparasites in El Valle de Anton, Cocle, Panama

The present research evaluated the presence of Rickettsia spp. on ectoparasites of horses and dogs (using PCR techniques), and their sera (using immunofluorescence assay) in El Valle de Anton town in Panama. A total of 20 horses and 20 dogs were sampled, finding four species of ectoparasites on dogs (the ticks Rhipicephalus sanguineus, Amblyomma ovale, Amblyomma oblongoguttatum, and the flea Ctenocephalides felis), and two tick species on horses (Amblyomma cajennense and Dermacentor nitens). DNA of Rickettsia amblyommii was found in pools of A. cajennense, D. nitens, and R. sanguineus, while Rickettsia fells was detected in C. felis pools. Overall, 70% (14/20) and 65% (13/20) of the horses and dogs, respectively, were seroreactive (titer >= 64) to spotted fever group rickettsiae. Sera from six dogs and five horses reacted to R. amblyommii antigens with titers at least four-fold higher than those for the other antigens tested (Rickettsia bellii, Rickettsia parked, Rickettsia rhipicephali, R. felis, and R. rickettsii). These serological results, coupled with our molecular findings, suggest that these dogs and horses were infected by Rickettsia amblyommii. More studies need to be realized afford to identify the Rickettsia species responsible for other serological and molecular positive results, and their ecological importance. (C) 2010 Elsevier B.V. All rights reserved.

Rickettsia in Synanthropic and Domestic Animals and Their Hosts from Two Areas of Low Endemicity for Brazilian Spotted Fever in the Eastern Region of Minas Gerais, Brazil

The aim of this study was to understand the current epidemiology of rickettsial diseases in two rickettsial-endemic regions in Brazil. In the municipalities of Pingo D`Agua and Santa Cruz do Escalvado, among serum samples obtained from horses and dogs, reactivity by immunofluorescent assay against spotted fever group rickettsiae was verified. In some serum samples from opossums (Didelphis aurita) captured in Santa Cruz do Escalvado, serologic response against rickettsiae was also verified. Polymerase chain reaction identified rickettsiae only in ticks and fleas obtained in Santa Cruz do Escalvado. Rickettsiae in samples had 100% sequence homology with Rickettsia fells. These results highlight the importance of marsupials in maintenance of the sylvatic cycle of rickettsial disease and potential integration with the domestic cycle. Our data also support the importance of horses and dogs as sentinels in monitoring circulation of rickettsiae in an urban area.

Wild canids, domestic dogs and their pathogens in Southeast Brazil: disease threats for canid conservation

Wild canids are under many pressures, including habitat loss, fragmentation and disease. The current lack of information on the status of wildlife health may hamper conservation efforts in Brazil. In this paper, we examined the prevalence of canine pathogens in 21 free-ranging wild canids, comprising 12 Cerdocyon thous (crab-eating fox), 7 Chrysocyon brachyurus (maned wolf), 2 Lycalopex vetulus (hoary fox), and 70 non-vaccinated domestic dogs from the Serra do Cip National Park area, Southeast Brazil. For wild canids, seroprevalence of antibodies to canine parvovirus, canine adenovirus, canine coronavirus and Toxoplasma gondii was 100 (21/21), 33 (7/21), 5 (1/19) and 68 (13/19) percent, respectively. Antibodies against canine distemper virus, Neospora caninum or Babesia spp. were not found. We tested domestic dogs for antibodies to canine parvovirus, canine distemper virus and Babesia spp., and seroprevalences were 59 (41/70), 66 (46/70), and 42 (40/70) percent, respectively, with significantly higher prevalence in domestic dogs for CDV (P < 0.001) and Babesia spp. (P = 0.002), and in wild canids for CPV (P < 0.001). We report for the first time evidence of exposure to canine coronavirus in wild hoary foxes, and Platynossomun sp. infection in wild maned wolves. Maned wolves are more exposed to helminths than crab-eating foxes, with a higher prevalence of Trichuridae and Ancylostomidae in the area. The most common ectoparasites were Amblyomma cajennense, A. tigrinum, and Pulex irritans. Such data is useful information on infectious diseases of Brazilian wild canids, revealing pathogens as a threat to wild canids in the area. Control measures are discussed.

Interface between domestic and wild carnivores in an environmental protection area in the brazilian Amazon: indicators and aplications for conservation

The presence of domestic animals in protected areas has been a growing world concern, including in the Brazilian Amazon. Introduced domestic carnivores can put the conservation of carnivores and other wild mammals at risk in protected areas. These risks can be associated with direct factors, such as the dogs` hunting habit, and indirect factors, such as the domestic carnivores` potential for transmitting infectious agents to wild populations. The objective of this study was to analyze the potential implications of humans and domestic animals staying in fragmented and altered areas such as the Wildlife Protection Areas (WPA) in the Tucuruf Lake Environmental Protection Area (Para, Brazil), created for the full protection of its fauna and flora. This evaluation was made through interviews conducted with riparian inhabitants living in the WPA and surrounding area, involving issues related to the presence of dogs (number of animals per domicile, birth rate, mortality rate, vaccination status, hunting habit) and wild carnivores (occurrence and location) in the study area. The results indicated risks mainly due to the presence of on average three dogs per domicile, as well as the high canine birth rate, their hunting habits, low vaccination rates and evidence of direct and indirect contact with the human, canine and wild carnivore populations. These factors represent risks to the health and conservation of Tucurui EPA wild carnivores since they provide favorable conditions for the transmission of pathogens from domestic fauna to wild fauna, as well as the risks brought about by hunting.

Serum Starvation and Full Confluency for Cell Cycle Synchronization of Domestic Cat (Felis catus) Foetal Fibroblasts

Nuclear transfer of domestic cat can be used as a tool to develop reproductive biotechnologies in wild felids. The importance of cell cycle phase during the nuclear transfer has been a matter of debate since the first mammalian clone was produced. The cell cycle phase of donor cells interferes on maintenance of correct ploidy and genetic reprogramming of the reconstructed embryo. The use of G0/G1 arrested donor cells has been shown to improve nuclear transfer efficiency. The present study was conducted to test the hypothesis that domestic cat foetal fibroblasts cultured up to the fifth passage and submitted to full confluency provide a higher percentage of cells at G0/G1 stage than fibroblasts cultured in serum starved media. Results demonstrated that serum starvation increased (p < 0.05) the percentage of G0/G1 fibroblasts when compared with control. Moreover, the combined protocol using confluency and serum starvation was more efficient (p < 0.05) synchronizing cells at G0/G1 stage than serum starvation or confluency alone for the first 3 days of treatment. In conclusion, serum starvation and full confluency act in a synergistic manner to improve domestic cat foetal fibroblast cell cycle synchronization at the G0/G1 stage.

Human directed aggression in Brazilian domestic cats: owner reported prevalence, contexts and risk factors

Aggression by cats towards humans is a serious behavioural, welfare and public health problem, although owners may believe it is an inevitable part of cat ownership. There has been little scientific investigation of the risk factors associated with this problem. One hundred and seven owners in the Sao Paulo region of Brazil, took part in a survey aimed at investigating the perceived prevalence of the problem, defining the most common contexts of human directed aggression and identifying associated potential risk factors. Human directed aggression occurred in 49.5%, of cats and was most commonly associated with situations involving petting and play, followed by protection of a resource, when startled, when observing an unfamiliar animal and least commonly when unfamiliar people were present. Pedigree status, neuter status, a history of early trauma, sensitivity to being stroked, the absence of other cats in the home, relationship with other animals, level of background activity at home, access to the outside and tendency to be alone (meaning tendency to staying far from the family members) were all associated with an increased risk in one or more context. However, sex, age, age when acquired, source of pet, attachment to a specific household member, type of domestic accommodation, relationship with another cat if present and contact with other animals did not appear to increase the risk. The results suggest sensitivity to being stroked and background levels of stress in the home are the most pervasive risk factors, and future research should aim to investigate these factors further. These data are of relevance when advising owners about the risk and development of this problem. (C) 2009 ESFM and AAFP. Published by Elsevier Ltd. All rights reserved.

Healing of critical-size cranial defects in guinea pigs using a bovine bone-derived resorbable membrane

Purpose: To investigate the healing of critical-size cranial bone defects (9-mm-diameter) in guinea pigs treated with a bovine bone-derived resorbable membrane. Materials and Methods: A sample of 42 guinea pigs was divided into test (n = 20), control (n = 20), and standard (n = 2) groups. A full-thickness trephine defect was made in the fronto-parietal bone of each animal. In the test group, the internal and external openings of the defect were each closed with a separate membrane, and the space between them was filled with blood clot and a central spacer. In the control group, the defect was filled only with the blood clot and spacer. At 1, 3, 6, and 9 months later, the calvarias (5 per period) for both the test and control groups were collected, fixed, radiographed, and histologically processed. The Standard-group animals were sacrificed immediately after surgery and used to determine the initial size of defect radiographically. The areas of defects in the radiographs were measured with image-analysis software and were compared between groups and periods by multiple regression analysis with the Bonferroni correction. Results: At 1 and 3 months, newly formed woven bone was histologically observed in both test and control groups. Radiographically, this new bone occupied an average of 32% of the defect area at 1 month and 60% at 3 months in the test group. In the control group, 21% of the defect was filled at 1 month and 39% at 3 months. However, the differences between treatments were not statistically significant (P > .05). At 6 and 9 months, a significant increase in newly formed lamellar bone was seen histologically in both groups. Radiographically, for the test group, the new bone occupied an average of 82% of the defect area at 6 months and 96% at 9 months. For the control group, new bone composed an average of 45% of the defect area at 6 months and 40% at 9 months. The differences between the test and control groups were statistically significant at 6 and 9 months (P < .05). Complete or almost complete filling of the defect was observed in several cases. Conclusion: It was concluded that the bovine bone-derived membrane is highly biocompatible and is able to promote good healing of critical-size defects in calvaria of guinea pig.