PESQUISA VETERINÁRIA BRASILEIRA

Abstract in English:

There are no studies that characterize the enteric nervous system (ENS) bats. The organization and density of myenteric neurons may vary according to the animal species, as well as the segment of the digestive tube considered. The nitric oxide is one of the key neurotransmitters present in the myenteric neurons, acting as a mediator in the smooth muscle relaxation. These neurons are evidenced by immunohistochemistry of nitric oxide synthase (NOS) or by NADPH-diaphorase histochemistry. In this sense, this study aimed to characterize the total neuronal population and subpopulation NADPH-d+ of the myenteric plexus present in the jejunum of the insectivore species Molossus rufus quantitatively. Five specimens were collected of M. rufus in a buffer area of the “Reserva Biológica das Perobas” in the microregion of Cianorte/PR. After the euthanasia, in a chamber saturated with isoflurane, segments were collected from the small intestine corresponding to the jejunum intended for two techniques for neuronal marking, Giemsa and NADPH-diaphorase, and a fragment to the histological technique of hematoxylin-eosin and Masson’s trichrome. All the procedures were approved by the “Comitê de Ética no Uso de Animais Unipar” (CEUA - protocol No. 34347/2017) and the “Instituto Chico Mendes de Conservação da Biodiversidade” (ICMBio - protocol No. 60061-1) The histological sections allowed to highlight the location of the myenteric plexus between the longitudinal and circular layers of the muscular tunic. The myenteric plexus had an average of total neuronal population (neurons Giemsa+) of 279.23 neurons/mm2, being the nitrergic neurons (neurons NADPH-d+) represented 20.4% of this total population, with an average of 58.14 neuron/mm2. Therefore, the collected data are consistent with previous studies in other mammalian species concerning the location of the myenteric plexus, as well as the neural myenteric proportion NADPH-d+ compared with the population of neurons Giemsa+. The gaps in the knowledge of ENS of bats limits comparative intraspecific and interspecific studies.

• Neurons myenteric plexus bats Molossus rufus Molossidae digestive tube enteric nervous system NADPH-diaphorase morphology

Abstract in Portuguese:

Não há estudos que caracterizem o sistema nervoso entérico (SNE) destes animais, configurando uma lacuna no conhecimento quanto à biologia destes indivíduos. A organização e densidade dos neurônios mientéricos podem variar de acordo com a espécie animal bem como o segmento do tubo digestório considerado. O óxido nítrico é um dos principais neurotransmissores presentes nos neurônios mientéricos, atuando como mediador no relaxamento do músculo liso gastrointestinal, de modo que estes neurônios são evidenciados igualmente pela imunohistoquímica da óxido nítrico-sintase (NOS) ou pela histoquímica da NADPH-diaforase. Neste sentido, objetivou-se caracterizar quantitativamente a população neuronal total e subpopulação NADPH-d+ do plexo mientérico presente no jejuno da espécie Molossus rufus de hábito alimentar insetívoro. Foram coletados cinco espécimes de M. rufus em área de amortecimento da Reserva Biológica das Perobas na microrregião de Cianorte/PR. Após a eutanásia, em câmara saturada com isoflurano, foram coletados segmentos do intestino delgado correspondentes ao jejuno destinados a duas técnicas para marcação neuronal, Giemsa e NADPH-diaforase e, um fragmento para a técnica histológica de hematoxilina-eosina e tricômio de Masson. Todos os procedimentos realizados foram aprovados pelo Comitê de Ética no Uso de Animais da Unipar (CEUA - protocolo nº 34347/2017) e pelo Instituto Chico Mendes de Conservação da Biodiversidade (ICMBio - protocolo nº 60061-1) Os cortes histológicos possibilitaram evidenciar a localização do plexo mientérico entre os estratos longitudinal e circular da túnica muscular. Neurônios Giemsa+ apresentaram uma média de 279,23 neurônios/mm2, já os neurônios nitrérgicos apresentaram em média 20,4% da população neuronal mientérica total, sendo evidenciados 58,14 neurônios NADPH-d+/mm2. Portanto, os dados coletados mostram-se condizentes com estudos anteriores em outras espécies de mamíferos quanto à localização do plexo mientérico, bem como, a proporção neuronal mientérica NADPH-d+ comparada com a população de neurônios Giemsa+. As lacunas existentes quanto ao conhecimento do SNE de morcegos limita possíveis inferências em comparativo intraespecífico e interespecífico.

• Neurônios plexo mientérico morcegos Molossus rufus molossídeos tubo digestório sistema nervoso entérico NADPH-diaforase morfologia

Abstract in English:

The Phyllostomidae family is important among the bats found in Brazil, with several species and diverse eating habits, and is the only one to have frugivorous representatives. These bats can be found in urban and in wild life environments in search for the best reproductive and feeding conditions. The versatility of environments can be associated with the incidence and/or distribution of some diseases through pathogenic agents. The present paper has the purpose to identify the oral and perianal microbiota and to detect the bacterial resistance of frugivorous bats captured near communities inhabited by humans in the northwestern region of the state of Paraná. A total of 68 bats were captured, belonging to four species of the Phyllostomidae family, namely Artibeus lituratus, Artibeus planirostris, Carollia perspicillata and Sturnira lillium, originated from forest fragments in the micro region of Umuarama, state of Paraná. A total of 64 isolates from oral bacteria and 39 from perianal region were submitted to identification. They were later submitted to a susceptibility test to 22 human and veterinary antimicrobials. The most prevalent bacteria were Escherichia coli 33.3% in the oral region, and 35.90% in the perianal region, Enterobacter aerogenes 12.7% and 5.13%, Enterobacter agglomerans 7.9% and 10.25%, and Serratia liquefaciens 9.5% and 5.13% in the oral and perianal region respectively. All bat species studied had resistant strains, with a few of them presenting multi-resistance to antimicrobials. The species with the highest multi-resistance index to antimicrobials was Carollia perspicillata, with three strains of the oral region resistant to 15 antimicrobials; it also presented two strains in the perianal region, which were resistant to 13 and 10 antimicrobials respectively. Based on the results found, it is possible to conclude that the oral and perianal microbiota of bats is composed of several enterobacterial species resistant to one or several antimicrobials used in human and veterinarian medicine. This is an issue and a future warning for unique health, since high percentages of resistance were found against antimicrobials broadly used, such as ampicillin, amoxicillin and amoxicillin+clavulonate.

• Bacterial resistance bats Phyllostomidae frugivorous bats gram-negative bacilli wild animals bacterioses

Abstract in Portuguese:

A família Phyllostomidae se destaca entre as famílias de morcegos encontrados no Brasil, com diversificadas espécies e hábitos alimentares, sendo a única a apresentar representantes frugívoros, podendo ser encontrada tanto em meio urbano, como de vida livre, em busca de melhores condições reprodutivas e alimentares. Essa versatilidade de ambientes pode estar associada à incidência e/ou distribuição de determinadas doenças por agentes patogênicos. O presente trabalho objetivou identificar a microbiota oral e perianal e detectar a resistência bacteriana em morcegos frugívoros capturados próximos às comunidades habitadas pelo homem na região noroeste do estado do Paraná. Foram capturados 68 morcegos, de quatro espécies da família Phyllostomidae, são eles Artibeus lituratus, Artibeus planirostris, Carollia perspicillata e Sturnira lillium, oriundos de fragmentos de Mata da microrregião de Umuarama, estado do Paraná. Um total de 64 isolados de bactérias da região oral e 39 da região perianal foram submetidos, identificação e posteriormente teste de susceptibilidade a 22 antimicrobianos de uso humano e veterinário. As bactérias mais prevalentes foram Escherichia coli 33,3% na região da boca e 35,90% na região perianal, Enterobacter aerogenes 12,7% e 5,13%, Enterobacter agglomerans 7,9% e 10,25% e Serratia liquefaciens 9,5% e 5,13% na região da boca e perianal, respectivamente. Todas as espécies de morcegos estudadas apresentaram cepas que foram resistentes, e algumas multirresistência aos antimicrobianos. A espécie que apresentou maior índice de multirresistência aos antimicrobianos foi Carollia perspicillata, com três cepas na região oral resistente a 15 antimicrobianos, e duas na perianal, com resistência a 13 e 10 antimicrobianos respectivamente. Baseados nos resultados encontrados, é possível concluir que a microbiota oral e perianal de morcegos, é composta por diversas espécies de enterobactérias, resistentes a um, ou vários antimicrobianos utilizados na medicina humana e veterinária, tornando-se um problema, e um alerta futuro para a saúde única, uma vez que foram encontrados elevados percentuais de resistência contra antimicrobianos utilizados em larga escala tais como ampicilina, amoxicilina e amoxicilina+clavulonato.

• Resistência bacteriana morcegos Phyllostomidae animais selvagens bacilos Gram-negativos filostomídeos morcegos frugívoros bacterioses

Abstract in English:

ABSTRACT.- Silverio S.M., Mari R.B, Clebis N.K., Scoz J.R., Germano R.M., Major J.A.A., Bombonato P.P. & Stabille S.R. 2009. The effects of ascorbic acid supplementation in ileum myenteric neurons of streptozotocin-induced diabetic rats. Pesquisa Veterinária Brasileira 29(4):295-302. Departamento de Cirurgia, Setor de Anatomia dos Animais Domésticos e Silvestres, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: remari@usp.br The exacerbation of the oxidative stress and of the polyol pathway which impair damage myenteric plexus are metabolic characteristics of diabetes. The ascorbic acid (AA) is an antioxidant and an aldose reductase inhibitor, which may act as neuroprotector. The effects of AA supplementation on the density and cellular body profile area (CP) of myenteric neurons in STZ-induced diabetes in rats were assessed. Four groups with five animals each were formed: normoglycemic (C); diabetic (D); AA-treated diabetic (DS) and AA-treated normoglycemic (CS). Dosagen of 50mg of AA were given, three times a week, for each animal (group DS and CS). Ninety days later and after euthanasia, the ileum was collected and processed for the NADPH-diaphorase technique. There were no differences (P>0.05) in the neuronal density among the groups. The CP area was lower (P<0.05) in the DS and CS groups, with a higher incidence of neurons with a CP area exceeding 200mm2 for groups C and D. The AA had no influence on the neuronal density in the ileum but had a neuroprotective effect, preventing the increase in the CP area and allowing a higher number of neurons with a CP area with less than 200mm2.

• Vitamin C diabetes intestine myenteric plexus

Abstract in Portuguese:

ABSTRACT.- Silverio S.M., Mari R.B, Clebis N.K., Scoz J.R., Germano R.M., Major J.A.A., Bombonato P.P. & Stabille S.R. 2009. The effects of ascorbic acid supplementation in ileum myenteric neurons of streptozotocin-induced diabetic rats. Pesquisa Veterinária Brasileira 29(4):295-302. Departamento de Cirurgia, Setor de Anatomia dos Animais Domésticos e Silvestres, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, São Paulo, SP 05508-270, Brazil. E-mail: remari@usp.br The exacerbation of the oxidative stress and of the polyol pathway which impair damage myenteric plexus are metabolic characteristics of diabetes. The ascorbic acid (AA) is an antioxidant and an aldose reductase inhibitor, which may act as neuroprotector. The effects of AA supplementation on the density and cellular body profile area (CP) of myenteric neurons in STZ-induced diabetes in rats were assessed. Four groups with five animals each were formed: normoglycemic (C); diabetic (D); AA-treated diabetic (DS) and AA-treated normoglycemic (CS). Dosagen of 50mg of AA were given, three times a week, for each animal (group DS and CS). Ninety days later and after euthanasia, the ileum was collected and processed for the NADPH-diaphorase technique. There were no differences (P>0.05) in the neuronal density among the groups. The CP area was lower (P<0.05) in the DS and CS groups, with a higher incidence of neurons with a CP area exceeding 200mm2 for groups C and D. The AA had no influence on the neuronal density in the ileum but had a neuroprotective effect, preventing the increase in the CP area and allowing a higher number of neurons with a CP area with less than 200mm2.

Abstract in English:

ABSTRACT.- Silverio S.M., Mari R.B., Clebis N.K., Scoz J.R., Germano R.M., Agreste F., Bombonato P.P. & Stabille S.R. 2008. Assessment of NADPH-diaphorase stained myenteric neurons of the jejunum of diabetic rats supplemented with ascorbic acid. Pesquisa Veterinária Brasileira 28(2):95-102. Departamento de Ciências Biológicas, UNIPAR, Campus-Paranavaí, Av. Huberto Brüning 360, Jardim Santos Dumont, Paranavaí, PR 87706-490, Brazil. E-mail: srstabille@wnet.com.br The relation between hyperglycemia and diabetic neuropathy has already been demonstrated in some studies. Among the theories proposed for its etiology the oxidative stress stands out. The performance of nitric oxide as a link between the metabolic and vascular neuropathogenic factors that triggers the diabetic neuropathy has already been put forward. This study aimed to assess the quantification and measurements of the cell body profile area (CBPA) of NADPH-diaphorase reactive (NADPH-dp) myenteric neurons of the jejunum of diabetic rats (induced by streptozotocin) supplemented with Ascorbic Acid (AA). These changes in the myenteric neurons seem to be related to the gastrointestinal disturbances observed in diabetes mellitus (DM). Twenty male Wistar rats (Rattus norvegicus) were distributed in 4 groups (n=5): controls (C), control supplemented (CS), diabetic (D), and diabetic suplemented (DS). DM was induced by estreptozotocin (50mg/kg body wt). One week after the induction and confirmation of the DM (glycemia exam), animals of the groups CS and DS received 50mg of AA three times a week by gavage. After 90 days of experiment, the animals were anesthetized with lethal thiopental dose (40mg/kg) and the collected jejunum processed for the histochemistry NADPH-diaphorase technique. Whole-mount preparations were obtained for quantitative and morphometric analysis of the myenteric neurons. A quantity of jejunum neurons in the Group D (96±7.5) was not different (P>0.05) from Group DS (116±8.08), C (92±9.7), and CS (81±5.4), but in Group DS the quantity was higher (P<0.05) than in Group C and CS. The CBPA of neurons from Group D (189.50±2.68µm2) and DS (195.92±3.75µm2) were lower (P<0.05) than from Group C (225.13±4.37µm2) and CS (210.23±3.15µm2). The streptozotocin-induced DM did not change the jejunum-ileum area, the jejunum myenteric plexus space organization and the density of NADPH-dp neurons. The 50g AA-supplementation, three times a week, during 90 days, did not decrease hyperglycemia; however, it had a neuroprotective effect on the myenteric neurons, minimizing the increase on the CBPA of NADPH-dp neurons and increasing the amount of NADPD-dp neurons.

• Myenteric plexus intestine vitamin C antioxidants hyperglycemia

Abstract in Portuguese:

ABSTRACT.- Silverio S.M., Mari R.B., Clebis N.K., Scoz J.R., Germano R.M., Agreste F., Bombonato P.P. & Stabille S.R. 2008. Assessment of NADPH-diaphorase stained myenteric neurons of the jejunum of diabetic rats supplemented with ascorbic acid. Pesquisa Veterinária Brasileira 28(2):95-102. Departamento de Ciências Biológicas, UNIPAR, Campus-Paranavaí, Av. Huberto Brüning 360, Jardim Santos Dumont, Paranavaí, PR 87706-490, Brazil. E-mail: srstabille@wnet.com.br The relation between hyperglycemia and diabetic neuropathy has already been demonstrated in some studies. Among the theories proposed for its etiology the oxidative stress stands out. The performance of nitric oxide as a link between the metabolic and vascular neuropathogenic factors that triggers the diabetic neuropathy has already been put forward. This study aimed to assess the quantification and measurements of the cell body profile area (CBPA) of NADPH-diaphorase reactive (NADPH-dp) myenteric neurons of the jejunum of diabetic rats (induced by streptozotocin) supplemented with Ascorbic Acid (AA). These changes in the myenteric neurons seem to be related to the gastrointestinal disturbances observed in diabetes mellitus (DM). Twenty male Wistar rats (Rattus norvegicus) were distributed in 4 groups (n=5): controls (C), control supplemented (CS), diabetic (D), and diabetic suplemented (DS). DM was induced by estreptozotocin (50mg/kg body wt). One week after the induction and confirmation of the DM (glycemia exam), animals of the groups CS and DS received 50mg of AA three times a week by gavage. After 90 days of experiment, the animals were anesthetized with lethal thiopental dose (40mg/kg) and the collected jejunum processed for the histochemistry NADPH-diaphorase technique. Whole-mount preparations were obtained for quantitative and morphometric analysis of the myenteric neurons. A quantity of jejunum neurons in the Group D (96±7.5) was not different (P>0.05) from Group DS (116±8.08), C (92±9.7), and CS (81±5.4), but in Group DS the quantity was higher (P<0.05) than in Group C and CS. The CBPA of neurons from Group D (189.50±2.68µm2) and DS (195.92±3.75µm2) were lower (P<0.05) than from Group C (225.13±4.37µm2) and CS (210.23±3.15µm2). The streptozotocin-induced DM did not change the jejunum-ileum area, the jejunum myenteric plexus space organization and the density of NADPH-dp neurons. The 50g AA-supplementation, three times a week, during 90 days, did not decrease hyperglycemia; however, it had a neuroprotective effect on the myenteric neurons, minimizing the increase on the CBPA of NADPH-dp neurons and increasing the amount of NADPD-dp neurons.