Innate immunity response of zafirlukast treated-tilapia during foreign body inflammation.

There is limited knowledge regarding the blockade of cysteinyl leukotriene receptors (CysLTRs) and their effects in teleost fish. The present study investigated the effects of Zafirlukast, antagonist of CysLTR1 receptor, on the foreign body inflammatory reaction in Nile tilapia (Oreochromis niloticus). Zafirlukast-treated tilapia demonstrated a decrease in the formation of multinucleated foreign body giant cells and Langhans cells on the round glass coverslips implanted in the subcutaneous tissue, along with a significant reduction in white blood cell counts and decreased production of reactive oxygen species. There was an increase in serum levels of α2-macroglobulins, as well as a decrease in ceruloplasmin and haptoglobin. Zafirlukast treatment led to a significant decrease in the area of splenic melanomacrophage centers and a reduction in the presence of lipofuscin. These findings highlight the potential anti-inflammatory effects of zafirlukast treatment in tilapia and indicate its action on CysLTR1 receptor, modulating the innate immune response of tilapia during the foreign body reaction. The comprehension of chronic inflammation mechanisms in fish has become increasingly relevant, especially concerning the utilization of biomaterials for vaccine and drug delivery.

Device implant based on poly (lactic acid) with vitamin E for vaccine delivery system in Tilapia: Study for biocompatibility and biodegradation.

The use of Poly (lactic acid) (PLA) as a slow-release vehicle for vaccines has attracted the attention of researchers, since its insertion improves the uptake of them, and reduces side effects or by stimulating recruited defense cells, assisting immunity without the need for booster vaccine doses. Seeking to develop new strategies for the administration of drugs and vaccines in aquaculture, we evaluated the biocompatibility and biodegradation of polymeric PLA devices and PLA plus vitamin E devices, implanted through subcutaneous (SC) and intraperitoneal (IP) routes in Nile tilapia. To carry out this study, 84 male tilapia (initial 243.82 ± 56.74 g; final 400.71 ± 100.54 g) were randomly distributed in 3 tanks (n = 28 fish per treatment/tank). The devices were prepared in two formulations: neat PLA (containing 100% PLA) and PLAVE (PLA plus vitamin E) implanted using a commercial AnimalTag® applicator, and non-implanted fish (control). Fish were sampled 15, 30, 60, and 120 days post-implantation (DPI). Blood analysis was used to access blood cells and blood smear for differential leucocytes count. Serum biochemistry to evaluated changes in serum proteins and glycemia. Histopathological investigation using hematoxylin-eosin (H&E) was used to assess polymer-tissue interaction. Histochemistry and immunohistochemistry was used to detection immune cells and phagocytes in capsule, and analyses of melanomacrophage centers (MMCs) to morphometric evaluation and percentage amount of melanin, hemosiderin and lipofucsin pigments. Histopathological study revealed an increase of capsular formation and inflammatory cell infiltration in PLAVE-implanted tilapia through SC route (15 DPI). Tilapia implanted with PLAVE and PLA (SC) presented mast cells and eosinophilic granular cells during 15, 30, and 60 DPI, with a decrease in these cells in the fibrous capsule around the polymer at 120 DPI. PLAVE implanted tilapia SC at 60 DPI showed significantly phagocytosis points than other groups. Phagocytic cells (F4/80+) were observed near to biopolymers in phagocytosis sites. Lipofuscin at 120 DPI in spleen melanomacrophage centers were significantly high in PLAVE implanted tilapias when compared to fish with PLA implants and control. The serum biochemical study of tilapia did not reveal changes in cytotoxicity and liver function in implanted fish. The absence of side effects in hematological and biochemical findings, including the absence of mortality after device implantation, proves its clinical safety. PLA implants in tilapia have demonstrated biocompatibility, biodegradation, clinical safety, and excellent evolution of foreign body inflammatory responses.

Meningitis Caused by Streptococcus agalactiae in Nile Tilapia (Oreochromis niloticus): Infection and Inflammatory Response.

Streptococcus agalactiae (Sta) of Lancefield group B is the primary etiological agent of bacterial meningitis in Nile tilapia and newborn humans. Thus, the study of this disease is of fundamental importance for aquaculture and human medicine. Additionally, elucidation of the mechanisms involved in the host-pathogenic response is important for the success of new therapies. In the present study, we elucidated important aspects of the innate immune response in the brain tissue of Nile tilapia (Oreochromis niloticus) infected by Sta. The neuroinflammatory process in the meninges started with the migration of MHC class II and CD68 + cells, production of TNF-alpha, and the effective immune response to Sta was mediated by the increased iNOs+. In conclusion, the present study brings a partial understanding of the pathophysiological and neuroinflammatory mechanisms in meningitis in Sta infected tilapia, enabling important advances in the therapy of this disease as well as the possibility of using this biological model to understand human meningitis.

Potential of mucoadhesive nanocapsules in drug release and toxicology in zebrafish.

Mucoadhesive polymeric nanocapsules have attracted interest of researchers from different fields from natural sciences because of their ability to interact with the mucosa and increase drug permeation. Anesthesia by immersion causes absorption through the skin and gills of fish, so it is important to evaluate the exposure of these organs to drug nanosystems. Benzocaine (BENZ) is one of the most popular anesthetic agents used in fish anesthesia, but it has drawbacks because of its low bioavailability, resulting in weak absorption after immersion. Here we describe method developed for preparing and characterizing chitosan-coated PLGA mucoadhesive nanoparticles containing BENZ (NPMAs) for zebrafish immersion anesthesia. We determined the lowest effective concentration, characterized the interaction of the mucoadhesive system with fish, measured the anesthetic efficacy, and evaluated possible toxic effects in embryos and adults exposed to the nanoformulations. This study opens perspectives for using nanoformulations prepared with BENZ in aquaculture, allowing reduction of dosage as well as promoting more effective anesthesia and improved interaction with the mucoadhesive system of fish.

Meningitis caused by Streptococcus agalactiae in nile tilapia (Oreochromis niloticus): infection and inflammatory response

Streptococcus agalactiae (Sta) of Lancefield group B is the primary etiological agent of bacterial meningitis in Nile tilapia and newborn humans. Thus, the study of this disease is of fundamental importance for aquaculture and human medicine. Additionally, elucidation of the mechanisms involved in the host–pathogenic response is important for the success of new therapies. In the present study, we elucidated important aspects of the innate immune response in the brain tissue of Nile tilapia (Oreochromis niloticus) infected by Sta. The neuroinflammatory process in the meninges started with the migration of MHC class II and CD68 + cells, production of TNF-alpha, and the effective immune response to Sta was mediated by the increased iNOs+. In conclusion, the present study brings a partial understanding of the pathophysiological and neuroinflammatory mechanisms in meningitis in Sta infected tilapia, enabling important advances in the therapy of this disease as well as the possibility of using this biological model to understand human meningitis.

Induced aerocystitis and hemato-immunological parameters in Nile tilapia fed supplemented diet with essential oil of Lippia alba / Aerocistite induzida e parâmetros hemato-imunológicos em tilápia do Nilo suplementada com óleo essencial de Lippia alba

The present study evaluated the dietary supplementation with essential oil of Lippia alba on the hemato-immunological parameters of Nile tilapia (Oreochromis niloticus) submitted to acute inflammation induced by carrageenin injection in the swim bladder. For a period of 45 days, 96 fish were divided in four treatments in triplicate, as follows: fish fed supplemented diet with essential oil of L. alba (4 mL kg-1 dry ration) injected with carrageenin; fish fed supplemented diet with cereal alcohol injected with carrageenin; fish fed unsupplemented diet with essential oil injected with carrageenin; fish fed unsupplemented diet and noninjected. Cortisol levels, erythrogram, leukogram and the inflammatory infiltrate were analyzed 6 h after inflammatory stimulus. Carrageenin-injected fish showed acute inflammatory reaction in the swim bladder characterized by higher infiltrate of neutrophils and monocytes. The circulating neutrophils number was significantly higher in fish fed L. alba when compared to other treatments. No difference in cortisol levels was found. For dose, time and administration form tested, supplementation with essential oil of L. alba did not present anti-inflammatory activity. On the other hand, influence of dietary supplementation was observed on the neutrophils number after induced aerocystitis highlighting its immunomodulatory characteristic.(AU)

O presente estudo avaliou a suplementação dietária com óleo essencial de Lippia alba sobre os parâmetros hemato-imunológicos em tilápias do Nilo (Oreochromis niloticus) submetidas à inflamação aguda induzida por carragenina na bexiga natatória. Pelo período de 45 dias, 96 peixes foram divididos em quarto tratamentos em triplicata: a) peixes suplementados com óleo esencial de L. alba (4 mL kg-1 de ração) injetados com carragenina; b) peixes suplementados com álcool de cereais injetados com carragenina; peixes não suplementados com óleo essencial injetados com carragenina; c) peixes não suplementados não injetados. Os níveis de cortisol, eritrograma, leucograma e o infiltrado inflamatório foram analisados seis horas após o estímulo inflamatório. Peixes injetados com carragenina apresentaram reação inflamatória aguda na bexiga natatória caracterizada por maior infiltrado de neutrófilos e monócitos. O número de neutrófilos circulantes foi significativamente maior nos peixes suplementados com L. alba quando comparado aos outros tratamentos. Não houve diferença nos níveis de cortisol. Para a dose, o tempo e a forma de administração testada, a suplementação com óleo essencil de L. alba não apresentou atividade anti-inflamatória. Por outro lado, foi constatada influência da suplementação dietária no número de neutrófilos após a aerocistite enfatizando a sua característica imunomoduladora.(AU)

Influence of dexamethasone and levamisole on macrophage recruitment, giant cell formation and blood parameters in the tropical fish Piaractus mesopotamicus / Influência da dexametasona e levamisol no recrutamento de macrófagos, formação de células gigantes e parâmetros hematológicos no peixe tropical Piaractus mesopotamicus

The aim of this study was to evaluate the effect of parenteral administration of dexamethasone and levamisole on the accumulation of macrophages and formation of giant cells in chronic inflammation by foreign body and blood parameters in pacu (Piaractus mesopotamicus). It was used 50 mg/kg of levamisole and 2.0 mg/kg of dexamethasone and the combination of both drugs. Coverslips were implanted under the skin. After 2, 7, and 15 days postimplantation (DPI), fish were anesthetized for removal of coverslips and the number of macrophages and giant cells were count. It was also collected blood from the caudal vessel to evaluate: red blood cell count, hematocrit, hemoglobin concentration, leukocytes count, thrombocytes count, MCV, and MCHC. We observed that dexamethasone affects negatively the formation of giant cells in the chronic inflammation for foreign body. Levamisole, despite being immunostimulatory in several species, showed limited action. However, it was enough to counteract the effect of dexamethasone; the association of the drugs did not interfere significantly in erythrocyte and leukocyte number in most of the treatments and times studied. In dexamethasone group there was a reduction in the number of erythrocytes and hemoglobin associated with increased mean corpuscular volume, suggesting slight macrocytic anemia. At 15 DPI, most groups showed the recovery of hematologic response. As in mammals, dexamethasone affects negatively the inflammatory response. Levamisole showed little effect by itself. However, in some parameters the association of both drugs causes similar response to control and naïve groups, showing the antagonistic effect of these drugs.

O objetivo deste estudo foi avaliar o efeito da administração parenteral de dexametasona e levamisol na acumulação de macrófagos e formação de células gigantes na inflamação crônica por corpo estranho e avaliação de parâmetros sanguíneos em pacu (Piaractus mesopotamicus). Utilizou-se 50 mg/kg de levamisol e 2,0 mg / kg de dexametasona e a combinação de ambos fármacos. As lamínulas foram implantadas sob a pele. Depois de 2, 7 e 15 dias pós-implantação (DPI), os peixes foram anestesiados para a remoção das lamínulas e contagem do número de macrófagos e células gigantes. Foi coletado sangue da veia caudal para realizar contagem de células vermelhas, leucócitos, trombócitos, concentração de hemoglobina, MCV e MCHC. Observou-se que a dexametasona afeta negativamente a formação de células gigantes na inflamação crónica por corpo estranho. O levamisol, apesar de ser considerado imunoestimulante em várias espécies, mostrou ação limitada. No entanto, foi suficiente para neutralizar o efeito da dexametasona; a associação das drogas não interferiu significativamente no número de eritrócitos e de leucócitos na maioria dos tratamentos e períodos estudados. No grupo da dexametasona, houve redução do número de eritrócitos e concentração de hemoglobina associado ao aumento do volume corpuscular médio sugerindo leve anemia macrocítica. Aos 15 DPI, a maioria dos grupos mostrou recuperação na resposta hematológica. Como em mamíferos, a dexametasona afeta negativamente a resposta inflamatória. O levamisol mostrou pouco efeito por si só. No entanto, em alguns parâmetros a associação das duas drogas provoca resposta similar ao grupo controle e naive, mostrando o efeito antagonista de estas drogas.

Eficiência do programa de higienização de uniformes em frigorífico / Efficiency of the hygiene program in a refrigerator

A qualidade da higienização industrial deve ser avaliada considerando- -se, entre outros aspectos, o risco que a contaminação alimentar representa para a saúde pública. Este trabalho buscou avaliar as condições higienicossanitárias de superfícies de 25 uniformes de frigorífico, por meio de análise microbiológica para micro-organismos mesófilos aeróbios (UFC/ cm²), conforme a técnica recomendada swab test, avaliando a eficiência do processo de lavagem dos uniformes. Observou-se que 4% das superfícies dos uniformes apresentaram contagens abaixo ou igual 4,0 x100 UFC/ cm², demonstrando condições higiênicas satisfatórias e indicando eficiência nas técnicas de higienização dos uniformes, portanto, não representando riscos de contaminação cruzada através dos uniformes. Tais achados demonstraram que o processo de higienização dos uniformes realizado pelo frigorífico é eficiente e eficaz na redução da carga microbiana, comprovando as boas condições higienicossanitárias dos mesmos.

Na2EDTA anticoagulant impaired blood samples from the teleost piaractus mesopotamicus / Anticoagulante Na2EDTA danifica amostras de sangue do teleósteo piaractus mesopotamicus

The present study aimed to evaluate the effects of Na heparin and Na2EDTA on blood of Piaractus mesopotamicus (360.7±42.4g, 26.4±1.0cm). Twenty fishes were sampled in two experiment trials, ten for erythrocyte fragility analysis and ten for hematologic and plasma biochemical study. The blood collected by venous-caudal puncture was fractioned and stored in anticoagulants solution: Na2EDTA 10%, Na2EDTA 3%, Na heparin 5000 IU and Na heparin 100 IU. Plasmatic levels of calcium presented in the Na2EDTA stored samples were about 80% lower than both heparin groups. Blood samples of P. mesopotamicus stored with Na2EDTA demonstrated increase in the hematocrit and MCV, and decrease in MCHC. The dose-response effect was observed in this study. The results are reinforced by the higher levels of plasmatic protein and hemolysis presented in the Na2EDTA 10% stored blood, confirming the deleterious effect of this anticoagulant treatment on the quality of blood samples. Na2EDTA is not indicated to store P. mesopotamicus blood samples, but sodium heparin at 100 IU is the most recommended anticoagulant, since this treatment presented the lower rate of alterations in the stored blood.

O presente estudo teve por objetivo avaliar os efeitos da heparina sódica e Na2EDTA no sangue de Piaractus mesopotamicus (360,7±42,4g, 26,4±1,0cm). Foram amostrados vinte peixes em dois ensaios experimentais, sendo dez peixes utilizados para análise da fragilidade dos eritrócitos e dez peixes para análise dos parâmetros hematológicos e estudo bioquímico do plasma. O sangue coletado por punção veno-caudal foi aliquotado e armazenado em diferentes soluções anticoagulantes: Na2EDTA 10%, Na2EDTA 3%, heparina sódica 5.000 UI e heparina sódica 100 UI. Níveis plasmáticos de cálcio apresentados nas amostras armazenados em Na2EDTA diminuíram cerca de 80% em relação aos dois grupos armazenados com heparina. Amostras de sangue de pacus armazenados com Na2EDTA demonstraram aumento do hematócrito e VCM, e diminuição na CHCM. O efeito dose-resposta foi observado neste estudo. Estes resultados são reforçados pelos níveis mais elevados de proteína plasmática e hemólise apresentado no sangue armazenado com Na2EDTA 10%, o que confirma o efeito deletério desse tratamento anticoagulante na qualidade de amostras de sangue. Na2EDTA não é indicada para armazenar amostras de sangue de P. mesopotamicus, e heparina sódica a 100 UI é o anticoagulante mais recomendado, uma vez que este tratamento apresentou a menor taxa de alterações no sangue armazenado.

Expression of cellular components in granulomatous inflammatory response in Piaractus mesopotamicus model.

The present study aimed to describe and characterize the cellular components during the evolution of chronic granulomatous inflammation in the teleost fish pacus (P. mesopotamicus) induced by Bacillus Calmette-Guerin (BCG), using S-100, iNOS and cytokeratin antibodies. 50 fish (120±5.0 g) were anesthetized and 45 inoculated with 20 µL (40 mg/mL) (2.0 x 10(6) CFU/mg) and five inoculated with saline (0,65%) into muscle tissue in the laterodorsal region. To evaluate the inflammatory process, nine fish inoculated with BCG and one control were sampled in five periods: 3rd, 7th, 14th, 21st and 33rd days post-inoculation (DPI). Immunohistochemical examination showed that the marking with anti-S-100 protein and anti-iNOS antibodies was weak, with a diffuse pattern, between the third and seventh DPI. From the 14th to the 33rd day, the marking became stronger and marked the cytoplasm of the macrophages. Positivity for cytokeratin was initially observed in the 14th DPI, and the stronger immunostaining in the 33rd day, period in which the epithelioid cells were more evident and the granuloma was fully formed. Also after the 14th day, a certain degree of cellular organization was observed, due to the arrangement of the macrophages around the inoculated material, with little evidence of edema. The arrangement of the macrophages around the inoculum, the fibroblasts, the lymphocytes and, in most cases, the presence of melanomacrophages formed the granuloma and kept the inoculum isolated in the 33rd DPI. The present study suggested that the granulomatous experimental model using teleost fish P. mesopotamicus presented a similar response to those observed in mammals, confirming its importance for studies of chronic inflammatory reaction.

Acute toxicity by water containing hexavalent or trivalent chromium in native Brazilian fish, Piaractus mesopotamicus: anatomopathological alterations and mortality.

This study evaluated the toxicity of hexavalent and trivalent compounds of chromium to the pacu, Piaractus mesopotamicus, in acute exposures of 96 h through mortality and histopathological responses. Hexavalent potassium dichromate was more toxic than trivalent compounds of chromium chloride, chromium oxide and chromium carbochelate. Sufficient mortalities occurred only with potassium dichromate to yield an LC50 value at 124.2 mg L(-1). Hexavalent chromium caused reversible and irreversible lesions, which may affect organ functionality. Histopathological evaluation showed that trivalent chromium caused lesions of lower severity. Pacu subjected to different concentrations of chromium carbochelate showed no histopathological changes in the kidneys, liver, skin and gills, being similar to those of the control fish. Among the three sources of Cr(3+), only chromium chloride at 200 mg L(-1) resulted in mortality, which reached 100 % within the first 18 h. These findings confirm that trivalent chromium, when administered within recommended levels, may be used safely in aquaculture.

Eficácia de diferentes formulações no controle da mosca Haematobia irritans em bovinos naturalmente infestados / Efficacy of different formulations on the control of Haematobia irritans in cattle naturally infested

Este estudo avaliou a eficácia antiparasitária da associação de cipermetrina (5%) + diclorvos (45%) + butóxido de piperonila (25%) contra H. irritans em bovinos naturalmente infestados. Para tal, foram utilizados 60 bovinos, fêmeas da raça nelore, com idade média de 50 meses, distribuídos em quatro grupos com 15 repetições cada. Após a randomização dos animais pelo número de moscas, os grupos foram sorteados e constituíram os seguintes tratamentos: T1 = Controle; T2 e T4= Tratados com a associação em estudo nas concentrações de 1:400 e 1:800, respectivamente; T3 = tratados com cipermetrina 15% +clorpirivos 25% + citronela 1% (Controle Positivo) na concentração de 1:800. As contagens de moscas foram efetuadas 3, 7, 14 e 21 dias após o tratamento (DPT). No terceiro DPT, os tratamentos T2, T3 e T4 apresentaram eficácia acima de 98%. No sétimo DPT, verificou-se nos animais tratados com a associação contendo butóxido de piperonila eficácia de 95,61 e 97,3% no controle de moscas nas doses de 1:800 e 1:400, respectivamente. Enquanto, bovinos do grupo T3 demonstraram eficácia de 93,82%. A eficácia antiparasitária permaneceu acima de 95% no 14° DPT em ambos os grupos T2 e T4. Neste mesmo dia experimental, bovinos do grupo T3 apresentaram eficácia de 88,24%. No 21° DPT, apenas o grupo T2 apresentou diminuição significativa (P<0,05) no número de moscas quando comparado aos demais tratamentos. Portanto, verificou-se neste estudo, o efeito sinergista do butóxido de piperonila sobre a atividade antiparasitária de piretróides e organofosforados, justificando o emprego terapêutico desta associação como uma alternativa eficaz em programas sanitários dos bovinos para controle da mosca dos chifres.

This study evaluated the effectiveness of cypermethrin (5%) associated to Dichlorvos (45%) and piperonyl butoxide (25%) against Haematobia irritans in naturally infested cattle. 60 Nelore females, aging about 50 months, were separated into four groups with 15animals each. After randomization by the number of flies, the groups were divided in the following treatments: T1 = control, T2 and T4 = treated with the association under study at concentrations of 1:400 and 1:800, respectively, T3 = treated with cypermethrin 15% + clorpirivos 25% + citronella 1% (positive control) at a concentration of 1:800. Counst of flies were taken 3, 7, 14 and 21 days post treatment (DPT). In the third DPT, treatments T2, T3 and T4 showed efficacy over 98%. At the seventh day, it was found in animals treated with the compound containing piperonyl butoxide efficacy of 95.61 and 97.3% in the control of flies at dose of 1:800 and 1:400, respectively. While cattle in the T3 group demonstrated efficacy of 93.82%. The effectiveness of antiparasitic remained above 95% at 14 ° DPT in both groups T2 and T4. In the same experimental day, animals of group T3 showed effectiveness of 88.24%. At 21 ° DPT, only the T2 group, was significantly decreased (P <0.05) in the number of flies when compared to other treatments. Therefore, it was found in this study, the synergistic effect of piperonyl butoxide on the antiparasitic activity of synthetic pyrethroids and organophosphates, representing an effective alternative in the chemotherapy control by the parasitism for horn flies in cattle.

Prilocaína a 3% associada à felipressina na anestesia peribulbar de gatos / Prilocaine 3% associated with felipressin peribulbar anesthesia in cats

O objetivo deste trabalho foi avaliar os efeitos da prilocaina associada à felipressina na anestesia peribulbar em gatos. Foram utilizados oito gatos adultos, machos e fêmeas, com peso entre 2 e 3 kg. Cada animal recebeu clorpromazina (1,0 mg.kg-1 por via intramuscular) como medicação pré anestésica e propofol (3,0 mg.kg-1 por via intravenoso) para indução anestésica. Foi introduzida uma agulha hipodérmica de 0,38 x 13 mm moldada manualmente até atingir a forma de "L" na porção superior da cavidade orbitária em direção ao fórnice e administrada prilocaína a 3% com vasoconstritor na dose de 5 mg.kg-1. Foi observada proptose após uma média de 2,28 ± 1,52 minutos, e este efeito durou em média 75,22 ± 26,99 minutos. Também ocorreu perda do reflexo corneano após 3,00 ± 1,69 minutos e sua duração foi de 73,75 ± 17,98 minutos em média. A prilocaína 3% com vasoconstritor manteve o globo ocular concêntrico, sem nistagmo e aboliu os reflexos corneanos sem causar efeitos colaterais.

The purpose of this study was to verify the effects of prilocaine 3% associated to felypressin for peribulbar anesthesia in cats. Eight adult cats, male and female, weighing between 2 and 3 kg, received chlorpromazine (1.0 mg.kg-1, intramuscular) and propofol (3.0 mg.kg-1, intravenously). While, prilocaine at 3% (5.0 mg.kg-1) was administered by using an L-shape hypodermic needle (0.38 x 13 mm), which was introduced in the upper portion of the orbital cavity towards the fornix. The onset of proptosis was observed in 2.28 ± 1.52 minutes and lasted 75.22 ± 26.99 minutes. The loss of the corneal reflex occurred after 3.00 ± 1.69 minutes and lasted 73.75 ± 17.98 minutes. It can be concluded that 3% prilocaine with felypressin solution induces centralization of the eyeball without nistagmus and abolition the corneal reflexes without causing any collateral effects in cats.

Avaliação de método digital para análise do índice funcional do ciático em ratos / Evaluation of digital method for analysis of the sciatic functional index in rats

Este trabalho teve como objetivo o estudo da reprodução e descrição de técnica para colheita digital de imagens durante a marcha de ratos e determinação do Índice Funcional do Ciático (IFC), através de uma passarela de vidro e obtenção de imagens com uma câmera digital. Após lesão por estrangulamento controlado de nervo ciático, na extensão de 3mm, durante 30 segundos, utilizando pinça hemostática, um grupo de 32 ratos foram amostrados 24h antes da lesão que serviu de controle e 24h, 7, 14 e 21 dias após a compressão do nervo ciático. Estes ensaios consistiam na filmagem da marcha de cada animal em vista inferior (através de um espelho a 45º), e posteriormente analisadas através do programa IMAGE-J. As medidas retiradas foram os comprimentos das patas (direita e esquerda posteriores), bem como, a distância entre os artelhos. Na análise do IFC, valores próximos do "zero" (0) sugerem que a função do nervo Ciático mantém-se preservada e valores próximos de "menos cem" (-100) indicam perda total de função. Verificou-se neste estudo que 24horas antes da cirurgia causadora da lesão, a média do IFC foi de -7,07 ± 7,88 e 24 horas após a lesão estes valores passaram para uma média de -77,95 ± 13,81, sendo cerca de 10 vezes maiores, em que 78% dos animais apresentavam de 60 a 100% de perda funcional na atividade motora que demonstrou recuperação gradativa ao longo dos dias analisados, comprovando a acuidade e eficácia da metodologia proposta. Tais resultados sugerem que estudos possam ser conduzidos de forma mais simples e barata com a técnica de obtenção e manipulação de imagens digitais das pegadas, durante a marcha de ratos em laboratório.

This work aimed to study the reproduction and description of technique for digital sampling images during rats gait and determination of the sciatic functional index (SFI), through a glass walkway to obtain shoots with a digital camera. After controlled injury by strangulation of sciatic nerve, with 3mm of length, during 30 seconds, using hemostatic forceps, a group of 32 rats was performed 24 hours before the lesion which served as control and 24 hours, 7, 14 and 21 days after injury. The tests consisted in the filming and shooting each animal in order to observe the view from below (by a mirror to 45°) and subsequently analyzed using the IMAGE-J program. Measures were taken from the lengths of the legs (right and left), and the distance between the ankle. In the analysis of IFC, values close to zero (0) suggest that the function of the sciatic nerve is still preserved and values coming from "less one hundred" (-100) indicate total loss of function. It was verified in this study that 24 hours before surgery the average SFI was -7.07 ± 7.88 and 24 hours after injury these values rose to an average of -77.95 ± 13.81, being about 10 times larger, where 78% of the animals showed 60 to 100% of functional loss in motor activity, demonstrated by the gradual recovery over the days analyzed, confirming the accuracy and effectiveness of the proposed methodology. These results suggest that studies can be conducted to simplify and reduce costs using the technique for digital images of footprints during rats gait in the laboratory.

Uso de ropivacaína a 0.5% em anestesia peribulbar de gatos / Use of 0.5% ropivacaine for peribulbar anesthesia in cats

Objetivou-se verificar a eficiência da ropivacaína na anestesia peribulbar em 10 gatos adultos sem raça definida, machos e fêmeas com peso variando de 2 a 3 kg. Os animais foram anestesiados com clorpromazina (1,0 mg/kg VO) e propofol (3,0 mg/kg IV). Foi introduzida uma agulha hipodérmica de 13 x 3,8 mm em forma de “L” na porção dorsal da cavidade orbitária em direção ao fórnice, e administrado 1,0 mg/kg de ropivacaína a 0,5%. Decorridos 10 minutos e um segundo observou-se proptose com duração de 100 minutos e seis segundos e perda do reflexo corneano após 10 minutos e cinco segundos, em média, com permanência de 124 minutos e dois segundos. O globo ocular manteve-se concêntrico por 121 minutos e cinco segundos, sem nistagmo. A ropivacaína a 0,5% no bloqueio peribulbar em gatos, ocasiona centralização do globo ocular e abole os reflexos corneanos.

The peribulbar anesthesia is largely used in corrective surgeries, such as glaucoma, cataract, and keratoplasty. The purpose of this study was to verify the efficacy of 0.5% ropivacaine for peribulbar anesthesia in cats. Ten adult cats, male and female, weighing between 2 and 3 kg, were used. The animals received chlorpromazine (1,0 mg/kg, orally) and propofol (3,0 mg/kg, intravenously). Ropivacaine at 0.5% (1,0 mg/kg) was administered by using a L-shape hypodermic needle (13x3.8mm), which was introduced in the upper portion of the orbital cavity towards the fornix. The onset of proptosis was observed in 10.1min and lasted 100.6min. The loss of the corneal reflex occurred after 10.5min and lasted 124.2min. The ocular globemaintained concentric for 121.5min and did not present nistagmus. It can be concluded that 0.5% ropivacaine solution induces centralization of the ocular globe, abolishes the corneal reflexes without causing any collateral effects in cats.