Management of urethral obstruction in a cat with epidural sacrococcygeal anesthesia guided by electrical neurostimulation

Background: The treatment for urethral obstruction in cats consists of catheterization, and for this, the cat must be sedated or anesthetized. Sacococcygeal epidural block has the advantage of being close to receptors related to nociception located in the spinal cord and it is safer because it represents lower risk of spinal cord injury or inadvertent application in the subarachnoid space, when compared to the lumbosacral epidural. Nerve stimulation through the neurolocator to identify the epidural space increases the accuracy of this technique. Thus, the objective is to report a case of epidural anesthesia with a sacrococcygeal approach guided by neurostimulation in a cat with urethral obstruction. Case: A 4-year-old male Siamese cat, weighing 4 kg, was referred to the veterinary care with a history of apathy and anorexia for 2 days. From the physical exam, the clinical diagnosis of urethral obstruction was made, and to desobstruction, we chose to perform sacrococcygeal epidural block. Initially, the patient was anesthetized with propofol (4 mg/kg) and midazolam (0.3 mg/kg). To perform the anesthetic block, the cat was placed in sternal decubitus with the hind limbs extended cranially to perform sacrococcygeal epidural block. The positive pole (cathode) was connected to the skin of the right inguinal region at the caudal aspect of the thigh and the neurostimulator was turned on and adjusted to 0.7 mA of stimulating current intensity, 0.1 ms duration and 1 Hz frequency. The needle for electrical neurolocation was introduced in the dorsal midline, perpendicular to the skin surface, between the spinous processes of S3-Cd1 in the skin. The exact injection point was obtained observed by the muscular response of the middle and distal third of the animal's tail with the neurostimulator adjusted to 0.3 mA of intensity, in the same duration and frequency as before. The total volume of 0.9 mL (0.22 mL/kg) of solution containing the combination of 0.6 mL of 0.75% levobupivacaine and 0.3 mL of 2% lidocaine was injected. The success of the block was confirmed by the loss of reflexes of the pelvic limbs and anal sphincter 10 min after the administration of the anesthetic solution. Discussion: In this case, the use of the neurolocator helped to perform an effective sacrococcygeal block, allowing urethral catheterization without the addition of other analgesic agents. This technique desensitizes and relaxes muscles in the regions of the perineum, anus, distal colon and penis, being useful for performing urethral catheterization. The use of smaller anaesthetic volumes to perform sacrococcygeal block makes it possible to achieve a more localized anesthesia, without affecting the motor function of the pelvic limbs. However, in our report, using a combination of levobupivacaine and lidocaine, the pelvic limb block was also verified despite the low volume applied. A hypothesis that could justify the different responses in relation to the pelvic limb block compared to other studies would be due to the different physicalchemical properties of the drugs used. Lidocaine is known to be less fat-soluble than bupivacaine, so it tends to spread more through the epidural space, in order to result in more extensive blocks. The use of a neurostimulator using a fixed electric current of 0.7 mA, pulse 0.1 ms and a frequency of 1 Hz allowed the correct identification of the needle position for the application of the anesthetic.

Thermal biology in river buffalo in the humid tropics: neurophysiological and behavioral responses assessed by infrared thermography

The present review aims to analyze the effect of extremely hot climates on the neurophysiological responses of thermal control and behavior in the river buffalo. Understanding thermal neuromodulation and its effects on the buffalo's behavior is of central importance, for this will allow us to make better decisions in terms of improving the level of welfare of buffaloes living in environments characterized by extreme heat, such as the humid tropics. The thermoregulation process involves a complex mechanism that begins with the integration of peripheral signals that are sent to the lateral parabrachial nucleus of the brainstem and then to the preoptic nucleus of the hypothalamus to generate physiological variations such as vasodilatation to dissipate heat under conditions of thermal stress, or vasoconstriction to conserve heat upon the perception of cold stimuli. The thermal biology of the river buffalo is based on several different adaptation mechanisms. The infrared thermography (IRT) technique will be of great help in this area because it permits the detection of vascular microcirculation changes in different thermal windows under adverse climatic conditions. Although buffaloes are classified as rustic animals, it is important to take into account their morpho-physiology and thermoregulation mechanisms to prevent thermal stress and the resulting poor welfare and reduced productivity. However, if appropriate thermoregulation facilities are provided (i.e., ponds, pools, potholes, or swampy areas), buffaloes can properly thermoregulate and tolerate high ambient temperatures. Therefore, they may represent a good option and an appropriate animal-based enterprise under climate change and global warming conditions.(AU)

Thermal biology in river buffalo in the humid tropics: neurophysiological and behavioral responses assessed by infrared thermography

The present review aims to analyze the effect of extremely hot climates on the neurophysiological responses of thermal control and behavior in the river buffalo. Understanding thermal neuromodulation and its effects on the buffalo's behavior is of central importance, for this will allow us to make better decisions in terms of improving the level of welfare of buffaloes living in environments characterized by extreme heat, such as the humid tropics. The thermoregulation process involves a complex mechanism that begins with the integration of peripheral signals that are sent to the lateral parabrachial nucleus of the brainstem and then to the preoptic nucleus of the hypothalamus to generate physiological variations such as vasodilatation to dissipate heat under conditions of thermal stress, or vasoconstriction to conserve heat upon the perception of cold stimuli. The thermal biology of the river buffalo is based on several different adaptation mechanisms. The infrared thermography (IRT) technique will be of great help in this area because it permits the detection of vascular microcirculation changes in different thermal windows under adverse climatic conditions. Although buffaloes are classified as rustic animals, it is important to take into account their morpho-physiology and thermoregulation mechanisms to prevent thermal stress and the resulting poor welfare and reduced productivity. However, if appropriate thermoregulation facilities are provided (i.e., ponds, pools, potholes, or swampy areas), buffaloes can properly thermoregulate and tolerate high ambient temperatures. Therefore, they may represent a good option and an appropriate animal-based enterprise under climate change and global warming conditions.

Eletroacupuntura na analgesia trans e pós-operatória de cadelas submetidas à ovariosalpingohisterectomia / Electroacupuncture in trans and postoperative analgesia in bitches submitted to ovariosalpingohisterectomy

Objetivou-se com este estudo avaliar o efeito analgésico trans e pós-operatório da eletroacupuntura em onda denso-dispersa e frequências 3 e 200Hz, nos pontos E44, R3 e BP4, compará-lo com a analgesia promovida pelos pontos BP6, E36 e VB 34, e pela morfina, em cadelas submetidas à ovariosalpingohisterectomia (OSH). Teve-se como hipótese que a eletroacupuntura nos pontos E44, R3 e BP4 resultaria em controle da dor trans e pós-operatória melhor ou igual àquele promovido pela eletroacupuntura nos pontos BP6, E36 e VB34 e pela morfina. Trinta e seis cadelas submetidas à cirurgia eletiva de OSH foram distribuídas em blocos ao acaso em três grupos com 12 animais. Em dois grupos foi realizada eletroacupuntura denso-dispersa, com frequência de 3 e 200Hz, sendo que, no primeiro grupo (GEA), foram estimulados os pontos BP4, E44, R3 e, no segundo grupo (GEB), os pontos BP6, E36, VB34. Nesses dois grupos, foram administrados 1,5mL de solução fisiológica por via IM. No terceiro grupo (GF), foi realizada acupuntura em três pontos falsos e o aparelho de eletroacupuntura foi acoplado, mas não foi ligado, e administrou-se morfina na dose de 0,5mg/kg, diluída em solução fisiológica de modo que o volume total foi de 1,5mL por via IM. Foram avaliados os parâmetros cardiovasculares e respiratórios durante o período pré e transoperatório. A avaliação de dor pós-operatória foi realizada durante 12 horas, e a analgesia suplementar com morfina foi realizada quando o escore de dor alcançou valores iguais ou superiores a nove pontos (total de 27) de acordo com a "Escala de dor da Universidade de Melbourne" (EDUM). O grupo GEB apresentou pressão arterial estável, menor variação da Etiso e menor requerimento analgésico no pós-operatório. Nos grupos GEA e GF houve aumento da Etiso e da pressão arterial nos momentos de maior manipulação cirúrgica. Os maiores escores de dor foram obtidos em MP0 (momento da extubação) no GEA (8) e em MP1(1h após a extubação) no GF (9)[...](AU)

Thirty-six dogs undergoing elective ovariohysterectomy surgery were randomly distributed into 3 groups of 12 animals each. In the first group, dense-dispersed electroacupuncture was performed with a frequency of 3-200 Hz in SP4, ST44, KID3 points and 1.5mL of saline was given intramuscularly (GEA group). For the second group, dense-dispersed electroacupuncture was performed with a frequency of 3-200 Hz in SP6, ST36, GB34 points and 1.5mL of saline given intramuscularly (GEB group). In the third group acupuncture was performed in 3 false points being the electroacupuncture device attached to the needles but not connected, and morphine was also administered at a dose of 0.5mg/kg diluted in saline so that the total volume was 1.5mL intramuscularly (GF group). Cardiovascular and respiratory parameters were evaluated during the pre and perioperative period. The evaluation of pain and postoperative sedation was performed for 12 hours and analgesic supplementation was performed whenever the pain score reached values above or equal to 9 points (in a total of 27) in accordance with the Scale of Pain of the University of Melbourne (EDUM). The GEB group had stable blood pressure and lower Etiso value as well as less need for additional analgesia postoperatively. The GEA group showed the highest values of Etiso and showed the greatest need for additional analgesia in the immediate postoperative period (at the moment of extubation). The GF group required more supplementary analgesia postoperatively and showed higher sedation scores up to 4 hours after surgery. The KID3, ST44 and SP4 acupoints used in electroacupuncture with frequency 3-200 Hz are not able to promote satisfactory antinociceptive action in the perioperative period. Postoperative analgesic effect, promoted by electrical stimulation in acupoints KID3, ST44 and SP4, is inferior to that provided by acupoints ST36, SP6 and GB34 and superior to that obtained with morphine(AU)

Perspectivas da aplicação das conotoxinas bloqueadoras de canais para cálcio dependentes de voltagem no trauma medular / Perspectives for the application of voltage-dependent calcium channels blockers conotoxin in spinal cord injury

O bloqueio dos canais para cálcio dependentes de voltagem é uma estratégia importante no tratamento do trauma medular, pois previne o influxo exacerbado do cálcio que participa ativamente em processos neurodegenerativos agudos, resultando em neuroproteção com melhora das funções neurológica. Dentre esses bloqueadores, as toxinas de caramujos marinhos são peptídeos com adequada estabilidade estrutural, estudadas pelas ações específicas em canais iônicos e receptores que interferem diretamente na liberação de neurotransmissores e na neuromodulação dos neurônios motores e sensitivos da medula espinal. Elas já são utilizadas no tratamento de desordens neurológicas e mostram-se promissoras no desenvolvimento de novas terapias para o trauma medular. Portanto, objetivou-se discorrer sobre a fisiopatologia do trauma medular e a possível utilização terapêutica das toxinas de caramujo marinho, atuantes nos principais canais para cálcio dependentes de voltagem.(AU)

Blocking voltage dependent calcium channels is an important strategy in acute spinal trauma treatment, because it prevents the exacerbated calcium influx which participates actively in acute neurodegenerative processes, resulting in neuroprotection with improvement of neurological and electrophysiological functions. The cone snail toxins are peptides with adequate structural stability, which have been studied by specific actions on ion channels and receptors that directly interfering in the release of neurotransmitters and neuromodulation of sensory and motor neurons of the spinal cord. They are already used in the treatment of neurological disorders and appear to be promising in the development of new therapies for spinal trauma. Therefore, it was aimed to discuss the pathophysiology of spinal cord trauma, and possible therapeutic use of marine snail toxins that acts in voltage-dependent calcium channels.(AU)

Electroanalgesia for the postoperative control pain in dogs / Eletroanalgesia para o controle da dor pós-operatória em cães

PURPOSE: To evaluate the analgesic and neuroendocrine effects of electroanalgesia in dogs undergoing ovariohysterectomy. METHODS: Eighteen dogs were randomly distributed to three groups of six animals each and received either electrical stimuli at acupuncture points (EA), at peri-incisional dermatomes (DER) and at both acupuncture points and peri-incisional dermatomes (EAD). Pre-anesthetic medication was acepromazine (0.05mg kg-1, IV). Anesthesia was induced with propofol (4 to 5mg kg-1, IV) and maintained with isoflurane. Postoperatively pain degree was measured using a numerical rating scale. Dogs were scored at 1, 3, 6, 12 and 24 hours postoperative. If the pain score was ≥6, supplemental morphine (0.5mg kg-1, IM) was administered. Serum cortisol concentration was measured before pre-anesthetic medication (basal), and at 1, 12 and 24 hours postoperative. RESULTS: EA and EAD- treated dogs had lower pain scores than DER treated dogs one hour postoperatively. Fewer EA and EAD-treated dogs required rescue analgesia. Serum cortisol did not differ among treatments. CONCLUSION: Preoperative application of electrical stimuli to acupuncture points isolated or in combination with peri-incisional dermatomes provides a reduced postoperative opioid requirement and promotes an effective analgesia in dogs undergoing ovariohyterectomy.(AU)

OBJETIVO: Avaliar os efeitos analgésicos e neuroendócrinos da eletroanalgesia em cadelas encaminhadas para ovariossalpingohisterectomia. MÉTODOS: Foram avaliadas 18 cadelas, distribuídas aleatoriamente em três tratamentos de seis animais cada: aplicação de estímulo elétrico em pontos de acupuntura (EA), em dermátomos periincisionais (DER) e em pontos de acupuntura associados aos dermátomos periincisionais (EAD). Todos os animais foram tranquilizados com acepromazina (0,05mg kg-1, IV), seguindo-se a indução com propofol (4 a 5mg kg-1, IV) e manutenção anestésica com isofluorano. O grau de analgesia foi avaliado 1, 3, 6, 12 e 24 horas após a cirurgia mediante escala descritiva numérica. Animais com escores ≥6 receberam analgesia de resgate com morfina (0,5mg kg-1, IM). A concentração sérica de cortisol foi avaliada antes da tranquilização, 1, 12 e 24 horas após a cirurgia. RESULTADOS: Escores inferiores de dor foram observados nos tratamentos EA e EAD em relação ao DER na primeira hora pós-operatória. A analgesia de resgate foi menos requerida nos animais dos tratamentos EA e EAD. O cortisol não diferiu entre os tratamentos. CONCLUSÃO: A eletroestimulação de acupontos isolados ou associados aos dermátomos periincisionais reduz o requerimento pós-operatório de opioides, bem como confere efetiva analgesia para cadelas submetidas à ovariossalpingohisterectomia.(AU)

Disfunção Sacrolíaca: Diagnóstico e reabilitação / Sacroiliac dysfunction: diagnosis and rehabilitation / Disfunción sacroilíaca: diagnóstico y rehabilitación

As afecções sacroilíacas são disfunções pélvicas bastante frequentes entre os cavalos de esporte,apesar de pouco diagnosticadas. Podem resultar em perda de desempenho, má postura, claudicações e relutância em executar exercícios específicos, impactando diretamente na vida atlética destes animais. Podes er causada por desmites dos ligamentos sacroilíacos ou atroses, com presença ou não de fragmentos ósseos.Seu diagnóstico é dependente de minunciosa inspeção estática, dinâmica, manobras específicas de mobilização pélvica e auxílios de imagem, dos quais os mais acessíves certamente são a termografia e o ultrassom. Seu tratamento deve ser multidisciplinar, com objetivos de resolução da dor, inflamação e reestabilização da musculatura pélvica intrínseca. Pode abranger estratégias como infiltrações com anti-inflamatórios esteroidais,ozônio, acupuntura, quiropraxia, eletroestimulação e exercícios de fortalecimento muscular. Relatou-se um caso de disfunção sacroilíaca causado por lascidão dos ligamentos sacroilíacos ventrais e atrose, tratado com sucesso durante um período de 60 dias com ozônio periarticular, quiropraxia, acupuntura e fortalecimento de músculos pélvicos e abdominais.

Sacroiliac disorders are quite frequent pelvic dysfunctions among sport horses, althoug hunder diagnosed. Can result in loss of performance, poor posture, lameness and reluctance to perform specific exercises, directly impacting the athletic life ofthese animais. It can be caused by desmites the sacroiliac ligamentsor atroses with presence or absence of bone fragments. Diagnosis is dependent on static and dynamic inspection,specific maneuvers of pelvic mobilization and imaging, of which the most frequently used are thermographyand ultrasound. The treatment should be multidisciplinary, with objectives of resolution of pain, inflammationand stabilization of intrinsic pelvic muscles. It may include strategies such as infiltrations with steroids, ozone,acupuncture, chiropractic, electrical stimulation and muscle strengthening exercises. We reported a case ofsacroiliac dysfunction caused by lascity of the ventral sacroiliac ligaments and athrosis successfully treated overa period of 60 days with periarticular ozone, chiropractic, acupuncture and strengthening of pelvic and abdominal muscles.

Trastornos sacroilíacas son disfunciones pélvicas bastante frecuentes entre los caballos de deporte, aunque infradiagnosticada. Puede resultar en pérdida de rendimiento, mala postura, Ia cojera y Ia renuencia arealizar ejercicios específicos, 10 que afecta directamente a Ia vida atlética de estos animales. Puede ser causadapor desmites los ligamentos sacroilíacas o atroses con presencia o ausencia de fragmentos de hueso. EI diagnóstico depende minunciosa inspección estática, maniobras específicas de movilización de Ia pelvis y Iaimagen dinámica de Ia ayuda, de los cuales Ia mayoría de los acessíves son ciertamente termografía y ultrasonido. Su tratamiento debe ser multi disciplinario, con los objetivos de Ia resolución dei dolor, Ia inflamación y Ia distensión músculos pélvicos intrínsecas. Puede incluir estrategias tales infiltraciones con fármacos anti inflamatorios noesteroideos, el ozono, Ia acupuntura, Ia quiropráctica, Ia estimulación eléctrica y ejercicios de fortalecimiento muscular. Reportamos un caso de disfunción sacroilíaca causada por lascidão los ligamentos sacroilíacasventral y Atrose tratado con éxito durante un período de 60 días con ozono periarticular, quiropráctica, acupunturay el fortalecimiento de los músculos pélvicos y abdominales.

Disfunção Sacrolíaca: Diagnóstico e reabilitação / Sacroiliac dysfunction: diagnosis and rehabilitation / Disfunción sacroilíaca: diagnóstico y rehabilitación

As afecções sacroilíacas são disfunções pélvicas bastante frequentes entre os cavalos de esporte,apesar de pouco diagnosticadas. Podem resultar em perda de desempenho, má postura, claudicações e relutância em executar exercícios específicos, impactando diretamente na vida atlética destes animais. Podes er causada por desmites dos ligamentos sacroilíacos ou atroses, com presença ou não de fragmentos ósseos.Seu diagnóstico é dependente de minunciosa inspeção estática, dinâmica, manobras específicas de mobilização pélvica e auxílios de imagem, dos quais os mais acessíves certamente são a termografia e o ultrassom. Seu tratamento deve ser multidisciplinar, com objetivos de resolução da dor, inflamação e reestabilização da musculatura pélvica intrínseca. Pode abranger estratégias como infiltrações com anti-inflamatórios esteroidais,ozônio, acupuntura, quiropraxia, eletroestimulação e exercícios de fortalecimento muscular. Relatou-se um caso de disfunção sacroilíaca causado por lascidão dos ligamentos sacroilíacos ventrais e atrose, tratado com sucesso durante um período de 60 dias com ozônio periarticular, quiropraxia, acupuntura e fortalecimento de músculos pélvicos e abdominais.(AU)

Sacroiliac disorders are quite frequent pelvic dysfunctions among sport horses, althoug hunder diagnosed. Can result in loss of performance, poor posture, lameness and reluctance to perform specific exercises, directly impacting the athletic life ofthese animais. It can be caused by desmites the sacroiliac ligamentsor atroses with presence or absence of bone fragments. Diagnosis is dependent on static and dynamic inspection,specific maneuvers of pelvic mobilization and imaging, of which the most frequently used are thermographyand ultrasound. The treatment should be multidisciplinary, with objectives of resolution of pain, inflammationand stabilization of intrinsic pelvic muscles. It may include strategies such as infiltrations with steroids, ozone,acupuncture, chiropractic, electrical stimulation and muscle strengthening exercises. We reported a case ofsacroiliac dysfunction caused by lascity of the ventral sacroiliac ligaments and athrosis successfully treated overa period of 60 days with periarticular ozone, chiropractic, acupuncture and strengthening of pelvic and abdominal muscles.(AU)

Trastornos sacroilíacas son disfunciones pélvicas bastante frecuentes entre los caballos de deporte, aunque infradiagnosticada. Puede resultar en pérdida de rendimiento, mala postura, Ia cojera y Ia renuencia arealizar ejercicios específicos, 10 que afecta directamente a Ia vida atlética de estos animales. Puede ser causadapor desmites los ligamentos sacroilíacas o atroses con presencia o ausencia de fragmentos de hueso. EI diagnóstico depende minunciosa inspección estática, maniobras específicas de movilización de Ia pelvis y Iaimagen dinámica de Ia ayuda, de los cuales Ia mayoría de los acessíves son ciertamente termografía y ultrasonido. Su tratamiento debe ser multi disciplinario, con los objetivos de Ia resolución dei dolor, Ia inflamación y Ia distensión músculos pélvicos intrínsecas. Puede incluir estrategias tales infiltraciones con fármacos anti inflamatorios noesteroideos, el ozono, Ia acupuntura, Ia quiropráctica, Ia estimulación eléctrica y ejercicios de fortalecimiento muscular. Reportamos un caso de disfunción sacroilíaca causada por lascidão los ligamentos sacroilíacasventral y Atrose tratado con éxito durante un período de 60 días con ozono periarticular, quiropráctica, acupunturay el fortalecimiento de los músculos pélvicos y abdominales.(AU)

Eletrophysiological study of the caudal nerve on developing rats / Estudo eletrofisiológico do nervo caudal em ratos em desenvolvimento

Purpose: To non-invasively study the sensory nerve conduction of the caudal nerve of normal developing rats. Methods: Twenty normal Wistar male rats served as subjects. Caudal nerve conduction studies were performed at 60 days from birth and weekly at end of six consecutive weeks. The caudal nerve was stimulated distally and nerve potentials were recorded proximally on the animal's tail using common "alligator" clips as surface electrodes. Results: The amplitude and the conduction velocity of the caudal nerve sensory action potential increased linearly from 29±6 µV to 85±13 µV and from 34±3 m/s to 44±4 m/s, respectively, between the 8th and the 15th week of life. The equations of linear regressions were as followed: Amplitude (µV) = 8.1 × weeks - 34 (R² = 0.99) and NCV (m/s) = 1.2 × weeks + 25 (R² = 0.86). Conclusions: It was possible to study non-invasively the sensitive conduction of the caudal nerve of normal developing rats and describe reference values. The technique and data may be used as animal model in physiological and pathological studies.(AU)

Objetivo: Estudar de forma não invasiva a condução nervosa sensitiva do nervo caudal em ratos normais em desenvolvimento. Métodos: Vinte ratos machos Wistar foram utilizados. A conducão nervosa foi realizada a partir dos 60 dias, semanal durante seis semanas consecutivas. O nervo caudal foi estimulado distalmente, os potenciais foram registrados proximalmente na cauda do animal usando garras comum do tipo "jacaré" como eletrodos de superficie. Resultados: A amplitude do potencial de ação de nervo sensitivo a velocidade de condução nervosa do nervo caudal cresceu linearmente de 29 ± 6 µV para 85 ± 13 µV e de 34 ± 3 m/s para 44 ± 4 m/s, respectivamente, entre a 8º e a 15º semana da vida do animal. As equações de regressão linear foram as seguintes: Amplitude (µV) = 8,1 × semanas - 34 (R² = 0,99) e velocidade de conducao nervosa (m/s) = 1,2 × semanas + 25 (R² = 0,86). Conclusões: Foi possível estudar de forma não invasiva a condução nervosa sensitiva do nervo caudal de ratos normais em desenvolvimento e descrever valores de referência. A técnica e os dados podem ser usados como modelo animal em estudos fisiológicos e patológicos.(AU)

Biomedical and pharmacological potential of tetrodotoxin-producing bacteria isolated from marine pufferfish Arothron hispidus (Muller, 1841)

Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55%, in Cellulomonas fimi with 620, 300, 10 and 128.42%, and in Bacillus species with 40, 200, 849.98 and 158.69%, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43% whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16% at the lowest dilution.Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55 percent, in Cellulomonas fimi with 620, 300, 10 and 128.42 percent, and in Bacillus species with 40, 200, 849.98 and 158.69 percent, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43 percent whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16 percent at the lowest dilution.(AU)

Biomedical and pharmacological potential of tetrodotoxin-producing bacteria isolated from marine pufferfish Arothron hispidus (Muller, 1841)

Specimens of the pufferfish Arothron hispidus collected at Parangipettai, on the southeast coast of India, were subjected to bacterial isolation and identification. Three species were identified, namely Bacillus sp., Kytococcus sedentarius and Cellulomonas fimi. Partially-purified microbial filtrates exhibited hemolytic activity on chicken and human erythrocytes of O, B and AB blood groups, with maximum activity of 32 HU. The microbial filtrates also presented ATPase, Mg2+-ATPase, Na+K+-ATPase and AchE enzymatic activities of positive neuromodulation in Kytococcus sedentarius with 1300, 300.1, 1549.98 and 140.55%, in Cellulomonas fimi with 620, 300, 10 and 128.42%, and in Bacillus species with 40, 200, 849.98 and 158.69%, respectively. Toxicity symptoms were observed when the bacterial filtrate was intraperitoneally injected into mice. The bacterial filtrate caused adverse effects on viability of the mouse muscle cell line (L929) and leukemia cell line (P388). Maximum level of inhibition was observed on the growth of L929 cell line. Bacillus lentimorbus inhibited the cell line from 84.03 to 94.43% whereas Bacillus species inhibited the growth in a range between 77.25 and 86.16% at the lowest dilution.(AU)

GnRH/GnIH e seus receptores no sistema olfatoretinal de zebrafish.

O hormônio liberador de gonadotropina (GnRH) é um dos fatores chaves na regulação neuroendócrina da reprodução dos vertebrados. Alguns peixes apresentam três variantes do GnRH: o GnRH1 envolvido na secreção de gonadotropinas, o GnRH2 que regula o comportamento alimentar e sexual e o GnRH3 expresso no bulbo olfatório e o nervo terminal cujas fibras nervosas inervam a retina e o epitélio olfatório. O zebrafish possui duas variantes do GnRH (GnRH2 e GnRH3), sendo o GnRH3 a variante hipofisiotrófica. Estudos mostram possível envolvimento do GnRH no sistema olfato-retinal. No sistema olfatório o GnRH regula a sensibilidade na detecção de alimento, o reconhecimento intra e interespecífico, entre outros. Na retina, o GnRH3 pode estar envolvido na acuidade visual e do processamento de informação da retina. Existem estudos que reportam a presença de receptores de GnRH em diferentes camadas da retina, no entanto ainda não é clara a presença de receptores no epitélio olfatório. Neste contexto, no presente estudo analisamos a localização do gnrh2, gnrh3 e seus receptores (gnrhr1,2,3 e 4) e do gnih (hormônio inibidor de gonadotropinas) no epitélio olfatório, a retina e o bulbo olfatório de machos e fêmeas adultos e comparamos a expressão destes genes em fêmeas em diferentes estágios de maturação gonadal. Para tanto, o RNA total do epitélio olfatório, retina, bulbo olfatório, cérebro e gônadas foi extraído. Com base na sequência dos genes gnrh2, gnrh3, gnrhr1, gnrhr2, gnrhr3 e gnrhr4, primers forward e reverse foram desenhados para RT-PCR e qPCR. Sondas para a hibridização in situ também foram construídas para verificar os sítios de expressão destas moléculas no epitélio olfatório, retina e gônadas. Imunohistoquímica com os anticorpos anti-GnRH3 (BB8 e GF6) foram realizadas para localizar a proteína do GnRH3 nos tecidos analisados. O presente estudo apresenta um panorama da expressão do sistema GnRH/GnIH e seus receptores no eixo epitélio olfatório-bulbo e olfatório-retina ao longo dos estágios de maturação gonadal em fêmeas de zebrafish. Os resultados mostram que no epitélio olfatório há um aumento na expressão dos receptores de GnRH nos estágios finais de maturação ovariana. O epitélio olfatório apresenta GnRH3 no citoplasma dos neurônios do receptor olfatório, assim como nos cílios destas células e também nas fibras do nervo olfatório. No bulbo olfatório, interessantemente, encontramos uma relação inversa entre a expressão de gnih e gnrh3 em todos os estágios de maturação gonadal. Na retina, os sítios de expressão do receptor gnrhr4 ocorrem nos núcleos das células amácrinas, bipolares e ganglionares, onde também foram observadas fibras positivas ao GnRH3. Nas gônadas femininas, o GnRH3 e seus receptores (gnrhr2 e gnrhr4) ocorrem em oócitos perinucleolares, enquanto que o gnih em oócitos em estágio de alvéolo cortical. Nas gônadas masculinas, o GnRH3 e seus receptores (gnrhr1 e gnrhr4) ocorrem em células germinativas (principalmente na gerações espermatogoniais), células de Sertoli e células de Leydig, enquanto que o gnih é amplamente expresso nas células germinativas e células intersticiais. O presente trabalho, de forma inédita, apresenta as variações na expressão gênica do sistema gnrh2, gnrh3, gnih, e das diferentes isoformas do receptor de GnRH (gnrhr1, gnrhr2, gnrhr3 e gnrhr4) nos três componentes do sistema olfato-retinal assim como nos ovários, sugerindo que o sistema GnRH/GnIH possa estar envolvido na neuromodulação dos sistemas sensoriais (olfatório e visual) especialmente nos estágios finais da maturação assim como a possível regulação dos processos de esteroidogênese e gametogênese nas gônadas do zebrafish.

The gonadotropin releasing hormone (GnRH) is one of the key factors involved in the neuroendocrine regulation of vertebrate reproduction. Some fish species have three GnRH variants: GnRH1 involved in gonadotropin secretion, GnRH2 regulating food and sexual behaviors and the GnRH3 which is expressed in the olfactory bulb and terminal nerve whose fibers innervate the retina and the olfactory epithelium. Two GnRH variants (GnRH2 and GnRH3) are present in the zebrafish, in which GnRH3 acts as the hypophisiotrophic variant. Recent studies have been showing the role of GnRH in the olfactory-retinal system. In the olfactory system, GnRH regulates food detection, and intra and interspecific recognition. In retina, GnRH3 may be involved in visual acuity modulation and retinal processing information. Moreover, studies have reported the presence of GnRH receptors in the retina, but not yet in the zebrafish olfactory epithelium. Therefore, the current study analyzed the presence of GnRH2, GnRH3 and its receptors (GnRH-R1,2,3 and 4) and GnIH (gonadotropin inhibitory hormone) in the olfactory epithelium, olfactory bulb, retina and in gonads of adult zebrafish. We also compared the expression of these genes during the different stages of ovarian maturation in zebrafish. For that, total RNA of the olfactory epithelium, olfactory bulb, retina and gonads was extracted with the PureLink® RNA Mini Kit(Ambion®). RT-PCR and qPCR analysis were performed using forward and reverse primers for gnrh2, gnrh3, gnrhr1, gnrhr2, gnrhr3, gnrhr4 for . Probes for in situ hybridization were constructed to verify the expression sites of these molecules in the olfactory epithelium, retina, and gonads. Immunohistochemistry usingantiGnRH3 antibodies (BB8 and GF6) were performed to identify the GnRH3 protein in these tissues. The current study presents a general expression view of GnRH/GnIH and their receptors in the olfactory epithelium-olfactory bulb-retinal axis during the female gonadal maturation in zebrafish. Our results showed an increase of GnRH receptor transcripts in the olfactory epithelium at the final stages of ovarian maturation. The olfactory epithelium exhibited GnRH3 in the cytoplasm of the olfactory receptor neurons, as well as the cilia of these cells and also in the olfactory nerve fibers. Interestingly, we found an inverse expression pattern for gnih and gnrh3 in the olfactory bulb, in all maturation stages. In the retina, gnrhr4 expression was found in the nuclei of amacrine, bipolar, and ganglion cells, where GnRH3 positive fibers could also be observed nearby to gnrhr4 expressing cells. In the female gonads, GnRH3 and their receptors (gnrhr2 and gnrhr4) were found in perinucleolar oocytes, whereas gnih in the cortical alveolus oocytes. In testis, GnRH3 and their receptors (gnrhr1 and gnrhr4) occur in germ cells (mainly in spermatogonial generations), Sertoli cells and Leydig cells, whereas gnih is widely expressed in germ cells and interstitial cells. This work describes for the first time the changes of gnrh2, gnrh3, gnih and the different GnRH receptor isoforms (gnrhr1, gnrhr2, gnrhr3, gnrhr4) expression in the three components of olfactory-retinal system, and in the ovary as well. Our results suggest that GnRH / GnIH system is involved in the neuromodulation of the sensorial system particularly at the final stages of maturation. We also address a possible role of GnRH/GnIH in steroidogenesis and gametogenesis processes in zebrafish gonads.

Bioactivities of extracts from the marine sponge Halichondria panicea

In the present study, we screened the biological activity of extracts from the marine sponge Halichondria panicea collected in the Arabian Sea. Crude toxin was obtained by methanol, chloroform-methanol (2:1) and aqueous extraction. Subsequently, the protein concentration of each crude extract was determined. The impact of both sponge methanolic and aqueous extracts was found to increase activities of Na+-K+ ATP-ase and Mg++ ATP-ase. In the case of chloroform-methanol extract, higher concentrations increased acetylcholine esterase (AchE) activity. The methanolic and chloroform-methanol extracts exhibited hemolytic activity on chicken and human erythrocytes, whereas the aqueous extract failed to do so. Methanol and aqueous extracts produced an immunostimulating effect and all extracts revealed angiogenic activity. The aqueous extract yielded nine bands by SDS-PAGE on 12% gel.

Bioactivities of extracts from the marine sponge Halichondria panicea

In the present study, we screened the biological activity of extracts from the marine sponge Halichondria panicea collected in the Arabian Sea. Crude toxin was obtained by methanol, chloroform-methanol (2:1) and aqueous extraction. Subsequently, the protein concentration of each crude extract was determined. The impact of both sponge methanolic and aqueous extracts was found to increase activities of Na+-K+ ATP-ase and Mg++ ATP-ase. In the case of chloroform-methanol extract, higher concentrations increased acetylcholine esterase (AchE) activity. The methanolic and chloroform-methanol extracts exhibited hemolytic activity on chicken and human erythrocytes, whereas the aqueous extract failed to do so. Methanol and aqueous extracts produced an immunostimulating effect and all extracts revealed angiogenic activity. The aqueous extract yielded nine bands by SDS-PAGE on 12% gel.(AU)

Effect of high frequency transcutaneous electrical nerve stimulation on viability of random skin flap in rats / Efeito da estimulação elétrica nervosa transcutânea em alta freqüência na viabilidade do retalho cutâneo randômico em ratos

OBJETIVO: Avaliar o efeito da Estimulação Elétrica Nervosa Transcutânea (TENS) em alta freqüência na viabilidade do retalho cutâneo randômico em ratos. MÉTODOS: Foram utilizados 75 ratos da linhagem Wistar. O retalho cutâneo apresentava 10 x 4 cm, sendo que entre o mesmo e a área doadora foi realizada a interposição de uma barreira plástica. Após o procedimento operatório todos os animais permaneceram anestesiados por mais uma hora com os eletrodos posicionados na base do retalho e submetidos ao tratamento de acordo com seus respectivos grupos. Este procedimento se repetiu nos outros dois dias subseqüentes. G1: simulação da TENS, G2: TENS (f = 80 Hz e I = 5 mA), G3: TENS (f = 80 Hz e I = 10 mA), G4: TENS (f = 80 Hz e I = 15 mA), G5: TENS (f = 80 Hz e I = 20 mA). RESULTADOS: A porcentagem média de área de necrose foi de 43,11, 34,65, 49,44, 23,52, 45,10 nos grupos 1, 2, 3, 4 e 5 respectivamente. CONCLUSÃO: Os animais estimulados com amplitude de 15 mA apresentaram menor área de necrose quando comparados ao grupo controle e a Estimulação Elétrica Nervosa Transcutânea foi eficiente no aumento de viabilidade do retalho cutâneo randômico em ratos.(AU)

PURPOSE: To determine the effect of high frequency Transcutaneous Electrical Nerve Stimulation (TENS) on viability of random skin flap in rats. METHODS: The sample of this study was 75 Wistar rats. The skin flap measured 10 x 4 cm and a plastic barrier was interposed between the flap and donor site. After the operative procedure, animals of all groups were maintained anesthetized one more hour with electrodes positioned in the base of the flap and submitted to treatment according of their respective group. This procedure was repeated on the two subsequent days. G1: sham stimulation (control), G2: TENS (f = 80 Hz and I = 5 mA), G3: TENS (f = 80 Hz and I = 10 mA), G4: TENS (f = 80 Hz and I = 15 mA), G5: TENS (f = 80 Hz and I = 20 mA). RESULTS: The average percentage of necrotic area was 43,11, 34,65, 49,44, 23,52, 45,10 in groups 1, 2, 3, 4 and 5 respectively. CONCLUSION: The amplitude of 15 mA presented a lower necrotic area than control group and Transcutaneous Electrical Nerve Stimulation was efficient in increasing the random skin flap viability.(AU)