Publications by authors named "Rafael Neodini Remedio"

15 Publications

  • Page 1 of 1

Repellent activity of acetylcarvacrol and its effects on salivary gland morphology in unfed Rhipicephalus sanguineus sensu lato ticks (Acari: Ixodidae).

Ticks Tick Borne Dis 2021 09 4;12(5):101760. Epub 2021 Jun 4.

Department of Health Sciences, Federal University of Lavras (UFLA), Av. Dr. Sylvio, Menicucci, 1001 Lavras, MG, Brazil. Electronic address:

Rhipicephalus sanguineus sensu lato (s.l.), commonly known as brown dog tick, is a widely distributed tick species that is substantially important for human and veterinary medicine. Therefore, it is the target of different control methods. Carvacrol and its semisynthetic derivative, acetylcarvacrol, are promising chemical compounds for alternative tick control. Thus, this study aimed to compare the repellent activities of carvacrol and acetylcarvacrol at different concentrations and drying times. Additionally, morphological alterations found in salivary glands were evaluated through histological techniques after exposure to acetylcarvacrol. The impact of the morphological changes on the development and survival of acini/cells in salivary glands was measured by a semiquantitative analysis. The repellent action of both compounds did not differ when evaluated at different concentrations, although acetylcarvacrol increased its effects as the concentration raised. Regarding the different drying times, acetylcarvacrol maintained its effects after 3 hours of exposure, while the efficacy of carvacrol decreased during this time period. Salivary glands of unfed R. sanguineus s.l. females showed dose-dependent alterations in the size and shape of acini as well as cytoplasmic vacuolization. Loss of the acinar cell limit, rupture of secretory granules and nuclear changes in the cells were also observed in the treated groups. Thus, our results demonstrated the potential of acetylcarvacrol to act as repellent against R. sanguineus s.l. Additionally, the morphological alterations found in salivary glands may interfere with the feeding process of ticks, which contributes to mitigate infestation by this species.
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http://dx.doi.org/10.1016/j.ttbdis.2021.101760DOI Listing
September 2021

Cytotoxic effects of Satureja montana L. essential oil on oocytes of engorged Rhipicephalus microplus female ticks (Acari: Ixodidae).

Microsc Res Tech 2021 Jul 6;84(7):1375-1388. Epub 2021 Jan 6.

Department of Health Sciences, Federal University of Lavras (UFLA), Lavras, Minas Gerais, Brazil.

In addition to pesticidal activity, the capacity of natural compounds to inhibit the reproduction of parasites emerge as an important alternative tick control method. In this context, Satureja spp. stand out due to their recognized pesticidal properties. Among parasites of veterinary importance, the cattle tick, Rhipicephalus microplus, is responsible for great economic losses in livestock and transmission of relevant pathogens. Thus, the aim of this study was to evaluate the effects of different concentrations of the essential oil of Satureja montana L. on the ovary morphology of R. microplus engorged females through histological and histochemical techniques. The most remarkable morphological changes found were: cytoplasmic vacuolation of germ cells, irregular and thicker chorion, irregular oocyte shape, ring-shaped nucleolus, decrease in protein and carbohydrate content in oocytes, in addition to cellular changes in the oviduct and pedicel. All morphological changes were assessed using a semiquantitative method already established in the literature. Ticks exposed to 5.0 μl/ml of essential oil showed the most significant changes when compared to control groups. Thus, the essential oil of S. montana L. damaged the reproductive system of R. microplus, which may impair ticks' offspring production and promote a long-term control of this species. HIGHLIGHTS: The essential oil of Satureja montana L. affects the ovary morphology of the cattle tick. The main morphological alterations found were cytoplasmic vacuolation, irregular and thicker chorion and irregular oocyte shape. These alterations may impair the development of eggs.
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http://dx.doi.org/10.1002/jemt.23693DOI Listing
July 2021

Sublethal concentrations of acetylcarvacrol affect reproduction and integument morphology in the brown dog tick Rhipicephalus sanguineus sensu lato (Acari: Ixodidae).

Exp Appl Acarol 2020 Oct 28;82(2):265-279. Epub 2020 Aug 28.

Department of Health Sciences, Federal University of Lavras (UFLA), Av. Dr. Sylvio Menicucci 1001, Lavras, MG, 37.200-900, Brazil.

Rhipicephalus sanguineus sensu lato (s.l.), also known as the brown dog tick, is among the main tick species involved in the transmission of pathogens to humans and other animals and, therefore, the target of numerous control methods. However, due to the disadvantages of synthetic acaricides, the use of alternative products such as plant derivatives has been encouraged. This study aimed to evaluate the acaricidal potential of acetylcarvacrol and to determine its efficacy at sublethal doses for the control of R. sanguineus s.l. female ticks. In addition, as acetylcarvacrol was applied topically, morphological alterations in the integument were assessed. Acetylation of carvacrol was performed by reaction with acetic anhydride in a sodium hydroxide solution, being confirmed by infrared spectroscopy. The lethal concentration for 50 and 90% (LC and LC) of unfed ticks and the efficacy of acetylcarvacrol in engorged females were determined after the Adult Immersion Test (AIT). For the evaluation of effects of acetylcarvacrol in the integument, routine histological techniques were employed after the AIT. The LC and LC in unfed females were 2.8 and 7.2 μL/mL, respectively. Regarding reproductive performance, after treatment with 8.0 μL/mL acetylcarvacrol 90.9% control was achieved, as ticks showed the lowest egg production index (EPI), hatching rate (HR), and fecundity rate (FR). In the integument, considerable morphological alterations were observed both in cuticle and epithelium. Thus, acetylcarvacrol affected R. sanguineus s.l. external coating and reproduction when applied at sublethal concentrations, probably contributing to a long-term control.
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http://dx.doi.org/10.1007/s10493-020-00538-7DOI Listing
October 2020

Acetylation of carvacrol raises its efficacy against engorged cattle ticks (Acari: Ixodidae).

Nat Prod Res 2020 Jun 27:1-5. Epub 2020 Jun 27.

Department of Health Sciences, Federal University of Lavras (UFLA), Lavras, MG, Brazil.

The effects of acetylcarvacrol on the reproductive performance of engorged female were evaluated. Carvacrol was acetylated by a reaction with acetic anhydride, confirmed by the identification of its melting point and by infrared spectroscopy. Based on the median lethal concentration, females were exposed to sublethal concentrations (3.7, 4.6 and 5.0 μL/mL) of acetylcarvacrol by means of the adult immersion test. The following parameters were evaluated: female weight before oviposition, pre-oviposition period, egg mass weight, egg production index, incubation period, hatching rate, fecundity rate, percentage of reduction in oviposition, percentage of reduction in hatching and product efficacy. The greatest efficacy was observed in the group treated with the highest concentration (91.69%) due to the greater weight reduction in egg mass (34.91 ± 0.02 mg) and the lower hatching rate (7.23 ± 15.50%). Therefore, this compound is a promising alternative for the control of infestations.
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http://dx.doi.org/10.1080/14786419.2020.1784169DOI Listing
June 2020

Low concentrations of acetylcarvacrol induce drastic morphological damages in ovaries of surviving Rhipicephalus sanguineus sensu lato ticks (Acari: Ixodidae).

Micron 2020 02 8;129:102780. Epub 2019 Nov 8.

Department of Health Sciences, Federal University of Lavras (UFLA), Lavras, MG, Brazil. Electronic address:

Rhipicephalus sanguineus sensu lato (s. l.) ticks are targets of acaricidal treatments due to their great veterinary and medical importance. Acetylcarvacrol stands out as a promising acaricidal substance for its increased biocidal activity and stability. Additionally, its toxicity to mammals is reduced when compared to the parent molecule, carvacrol. The present study aimed to evaluate the effects of acetylcarvacrol on the morphology of ovaries of engorged R. sanguineus s. l. ticks. The animals were subjected to the Adult Immersion Test to calculate the lethal concentrations (LC and LC) of acetylcarvacrol. Subsequently, the surviving ticks were dissected for collection of the ovaries. The samples were processed through routine histological techniques. The histological sections were stained with hematoxylin and eosin, bromophenol blue (protein detection) and PAS (polysaccharide detection). The alterations found in the ovaries were evaluated by morphometric and semiquantitative analysis. The LC and LC were 17.805 and 26.164 μL/mL, respectively. The most severe morphological alterations were disappearance of the female germ cells (oocytes), nucleolus vacuolization, thicker and irregular chorion, and decrease in size of the oocytes. Also, the content of proteins and carbohydrates in the oocytes were heavily affected by the chemical, as evidenced by a non-homogeneous staining pattern. The group exposed to the highest concentration of acetylcarvacrol (20 μL/mL) exhibited a statistically greater score in the semiquantitative analysis when compared to the other groups. The morphological changes in the ovaries may reduce the ticks' offspring production or generate descendants that will struggle to carry out essential biochemical processes during their lives. Thus, acetylcarvacrol may be a promising alternative to control tick infestation by impairing the reproduction of this parasite.
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http://dx.doi.org/10.1016/j.micron.2019.102780DOI Listing
February 2020

Effects of carvacrol on oocyte development in semi-engorged Rhipicephalus sanguineus sensu lato females ticks (Acari: Ixodidae).

Micron 2019 01 29;116:66-72. Epub 2018 Sep 29.

Department of Biology, Institute of Biosciences, São Paulo State University - UNESP, Rio Claro, SP, Brazil. Electronic address:

Currently, the most commonly used method to control ticks is the use of synthetic acaricides. However, these compounds are potentially harmful to hosts and the environment, in addition to causing the selection of resistant individuals. Therefore, several studies have been conducted to find sustainable methods to control ticks, such as Rhipicephalus sanguineus sensu lato, the most important vectors of pathogens for dogs. There has been increasing research on natural products with acaricidal action, especially with regard to plant-derived compounds as carvacrol, an aromatic monoterpene with several biological properties, including repellent and acaricidal activities, besides exerting cytotoxic effects on the exposed ticks. The objective of the present study was to evaluate the action of different carvacrol concentrations on the ovaries morphophysiology in semi-engorged R. sanguineus s.l. females to determine its effect on oocyte development. The results showed the occurrence of significant morphological alterations in the shape of oocytes (from round-shaped to irregular) and in the germinal vesicles, in addition to extensive cytoplasmic vacuolation.These effects were observed after the application of carvacrol at a concentration of 20 μL/mL. The most significant alterations were observed at the highest concentration (100 μL/mL), at which the oocytes could not develop further than stage II (total absence of oocytes III, IV and V). These data showed that even though carvacrol was unable to kill all ticks at these concentrations, surviving females could have had an altered reproduction, which would hinder the generation of new individuals, resulting in a long-term control. Data regarding the inhibition of oocyte development are unprecedented and indicate the use of carvacrol as a natural product with the potential to control R. sanguineus s.l. ticks.
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http://dx.doi.org/10.1016/j.micron.2018.09.015DOI Listing
January 2019

The effects of neem oil (Azadirachta indica A. JUSS) enriched with different concentrations of azadirachtin on the integument of semi-engorged Rhipicephalus sanguineus sensu lato (Acari: Ixodidae) females.

Microsc Res Tech 2017 Aug 29;80(8):838-844. Epub 2017 Mar 29.

Department of Biology, Institute of Biosciences, São Paulo State University, UNESP, Av. 24 A, no 1515, CP 199, Rio Claro, SP, 13506-900, Brazil.

Several studies searching for methods to control Rhipicephalus sanguineus s.l., (dog tick) infestations have been developed aiming to minimize the damages caused by these ectoparasites to the hosts and the environment, which is harmed by the indiscriminate use of toxic acaricide products. In this scenario, neem oil has been used as a natural alternative against ticks, once this chemical has repellent properties and interferes in the growth regulation of these ectoparasites, inhibiting ecdysis. The present study evaluated the effects of azadirachtin-enriched neem oil on the integument of semi-engorged R.sanguineus s.l., females through morphohistological techniques. The results showed the occurrence of significant morphological and histochemical alterations, mainly in the females exposed to higher concentrations, which demonstrates the dose-dependent action of the chemical. A decrease in the cuticle thickness was observed, as well as a modification in the distribution of the epithelial cells, which displayed pyknotic and fragmented nuclei, and intensely vacuolated cytoplasm, indicating that these cells would be undergoing death processes. These morphological alterations observed in the integument of the females exposed to the azadirachtin-enriched neem oil encourage the use of this chemical as a strategy to control these ectoparasites.
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http://dx.doi.org/10.1002/jemt.22871DOI Listing
August 2017

Dinotefuran-induced morphophysiological changes in the ovaries and midgut of semi-engorged females Rhipicephalus sanguineus Latreille, 1806 (Acari: Ixodidae) ticks.

Parasitol Res 2016 Feb 28;115(2):829-49. Epub 2015 Nov 28.

Department of Biology, Institute of Biosciences, São Paulo State University-UNESP, Av. 24 A, n° 1515, Postal Code 199, Zip Code: 13506-900, Rio Claro, SP, Brazil.

The present study demonstrated the effects of dinotefuran (active compound of the Protetor Pet® acaricide) in germ cells and the digestive processes of semi-engorged females of R. sanguineus exposed to different concentrations of the chemical. For this purpose, 120 semi-engorged females were divided into four treatment groups with 30 individuals each: group I or control (distilled water), group II (5000 ppm), group III (6250 ppm), and group IV (8334 ppm of dinotefuran). All ticks were immersed in different concentrations of dinotefuran or in distilled water for 5 min and then were dried and stored in biological oxygen demand (BOD) incubator for 7 days. The results show the action of this compound, exhibiting morphohistologic and histochemical changes in the oocytes and the midgut cells of individuals of different groups, which were compared with those of group I (control). The alterations occurred mainly in relation to the size of the germ cells and yolk granules; presence, quantity, size, and location of vacuoles found in the cytoplasm of these germ cells; the damage occurred in the generative cells of the midgut; the size of the digestive cells; the quantity of blood elements captured, accumulated digestive wastes and digestive vacuoles found in the cytoplasm of the digestive cells of the midgut, as well as the amount and distribution of proteins, polysaccharides, lipids of all cells in both organs. So, it has demonstrated the effectiveness of dinotefuran in the reduction of fertility and digestive processes of semi-engorged females of R. sanguineus, data that points the possibility of employing this chemical to control these ectoparasites.
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http://dx.doi.org/10.1007/s00436-015-4814-3DOI Listing
February 2016

Potential of the chemical dinotefuran in the control of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae) semi-engorged female ticks.

Exp Parasitol 2015 Aug 5;155:82-8. Epub 2015 May 5.

Department of Biology, Institute of Biosciences, São Paulo State University-UNESP, Av. 24 A, n° 1515, 199, 13506-900, Rio Claro, SP, Brazil.

Ticks are vectors of several pathogens to vertebrates, including the human being. They produce lesions on the hosts during the blood feeding and great economic losses. Several chemical acaricides have been used in an attempt to control tick infestations; however these substances are harmful to both the human being and non-target organisms, and to the environment. Therefore, there is a need to fight these ectoparasites through less harmful methods, less aggressive to the environment, non-target organisms and to the human health. The present study examined the efficacy of dinotefuran on the susceptibility of Rhipicephalus sanguineus semi-engorged females exposed to different concentrations of the product. Its lethal concentration of 50% (LC50) at 95% confidence interval was determined. The ticks were immersed in Petri dishes containing different concentrations of dinotefuran or distilled water for 5 minutes and then dried and maintained in an incubator for 7 days. The results showed the daily number of dead R. sanguineus semi-engorged females after being treated with different concentrations of dinotefuran. The mortality data in bioassay 2 were subjected to Probit analysis, where a LC50 of 10,182.253 ppm (8725.987-13,440.084) and 95% confidence interval were estimated. The susceptibility of R. sanguineus semi-engorged females to dinotefuran in higher concentrations of the acaricide was demonstrated, indicating that its effect is probably dose-dependent. In addition, the action of dinotefuran was slow and gradual, interfering in the development and growth of the individuals throughout the observation period (7 days).
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http://dx.doi.org/10.1016/j.exppara.2015.04.021DOI Listing
August 2015

Histopathological study of ovaries of Rhipicephalus sanguineus (Acari: Ixodidae) exposed to different thymol concentrations.

Parasitol Res 2014 Dec 10;113(12):4555-65. Epub 2014 Oct 10.

Programa de Pós-graduação em Comportamento e Biologia Animal, Universidade Federal de Juiz de Fora, Rua José Lourenço Kelmer, s/n - Campus Universitário Bairro Martelos, 36036-330, Juiz de Fora, MG, Brazil,

Thymol is a monoterpene with proven acaricide action for several tick species. In addition to killing these ectoparasites, thymol can also reduce oviposition and egg hatch rate. However, the effects of thymol on the morphophysiology of tick ovaries are still unknown. Thus, the aim of this study was to evaluate the morphophysiological changes caused by this active principle in ovaries of Rhipicephalus sanguineus after a 6-day feeding period, through the application of morphohistochemical techniques. After the feeding period, a total of 50 females were divided into five groups and immersed in the following solutions: (I) distilled water (control), (II) 30% ethanol (control), (III) 1.25 mg/mL thymol, (IV) 2.5 mg/mL thymol, and (V) 5.0 mg/mL thymol. The experimental groups were kept in a climatic chamber (27 ± 1 °C; RH 80 ± 10%) for 5 days. After this period, morphological (hematoxylin/eosin) and histochemical (von Kossa) techniques were applied after remotion of the ovaries. The morphological results revealed large vacuoles in germ cells at different developmental stages and invaginations that represent deformations in the chorionic membrane. From the results obtained in this study, it was concluded that thymol interfered with the development of oocytes, which showed degeneration signs. The treatment containing 5.0 mg/mL thymol affected more accentuately the morphological development. Moreover, thymol also altered the calcium content of yolk granules, which generally showed an intense staining for this element.
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http://dx.doi.org/10.1007/s00436-014-4145-9DOI Listing
December 2014

Effects of andiroba (Carapa guianensis) oil in ticks: Ultrastructural analysis of the synganglion of Rhipicephalus sanguineus (Latreille, 1806) (Acari: Ixodidae).

Acta Trop 2015 Jan 26;141(Pt A):7-15. Epub 2014 Sep 26.

UNESP-Universidade Estadual Paulista, Instituto de Biociências, Departamento de Biologia, Avenida 24 A, 1515, Bela Vista, Rio Claro CEP 13506-900, SP, Brazil.

The present study performed an analysis of the ultrastructural changes induced by andiroba seed oil in the synganglion of Rhipicephalus sanguineus female ticks, aiming to provide scientific grounds to help in the creation of more specific and efficient methods of control. The synganglion consists of a mass of fused nerves externally covered by the neural lamella, a uniform and acellular layer. Just below, the perineurium is found, formed by glial cells. Internally, the synganglion is subdivided into an outer cortical region (cortex), which contains the cellular bodies of the neural cells and an inner region, the neuropile, formed by a set of nerve fibers (extensions of the neural cells). The results showed that the synganglion of females exposed to andiroba oil showed structural changes, such as: irregular and apparently thinner neural lamella, perineurium glial cells presenting large cytoplasmic vacuoles, decrease in the extensions of glial cells, separation of cortex cells, which were formerly attached through their membranes, neural cells presenting irregular plasma membranes and cytoplasm with autophagic vacuoles and mitochondria with disorganized cristae and in process of degeneration. This study confirmed the neurotoxic action of the andiroba oil, which would probably be able to impair the neural functions. Thus, it is suggested that this product has the potential to be used as an alternative method to control ticks.
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http://dx.doi.org/10.1016/j.actatropica.2014.06.018DOI Listing
January 2015

Central nervous system of Rhipicephalus sanguineus ticks (Acari: Ixodidae): an ultrastructural study.

Parasitol Res 2012 Sep 19;111(3):1277-85. Epub 2012 May 19.

Departamento de Biologia, Instituto de Biociências, Universidade Estadual Paulista Júlio de Mesquita Filho, UNESP, Avenida 24 A, 1515, 13506-900, Rio Claro, SP, Brazil.

This study performed the ultrastructural description of the synganglion of Rhipicephalus sanguineus males and females, aiming to contribute to the understanding of the cellular organization of this organ. The results show that the central nervous system of these individuals consists of a mass of fused nerves, named synganglion, from where nerves emerge towards several parts of the body. It is surrounded by the neural lamella, a uniform and acellular layer, constituted by repeated layers of homogeneous and finely granular material. The perineurium is just below, composed of glial cells, which extensions invaginate throughout the nervous tissue. The synganglion is internally divided into an outer cortex, which contains the cellular bodies of the neural cells and an inner neuropile. The neural cells can be classified into two types according to cell size, cytoplasm-nucleus relation, and neurosecretory activity. Type I cells are oval or spherical and present a large nucleus occupying most part of the cytoplasm, which contains few organelles. Type 2 cells are polygonal, present a great cytoplasm volume, and their nuclei are located in the cell periphery. The cytoplasm of these cells contains a well-developed rough endoplasmic reticulum, Golgi regions, mitochondria, and several neurosecretory granules. The subperineurium and the tracheal ramifications are found between the cortex and the neuropile. The latter is formed mainly by neural fibers, tracheal elements, and glial cells. The results obtained show that R. sanguineus males' and females' nervous tissue present an ultrastructural organization similar to the one described in the literature for other tick species.
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http://dx.doi.org/10.1007/s00436-012-2962-2DOI Listing
September 2012

Synganglion histology in different stages of Rhipicephalus sanguineus ticks (Acari: Ixodidae).

Parasitol Res 2012 Jun 5;110(6):2455-63. Epub 2012 Jan 5.

Departamento de Biologia, Instituto de Biociências, Universidade Estadual Paulista "Júlio de Mesquita Filho", UNESP, Avenida 24 A, 1515, 13506-900, Rio Claro, São Paulo, Brazil.

The present study performs a morpho-histological description of the central nervous system of Rhipicephalus sanguineus ticks in order to analyze if there are differences in this tissue among larvae, nymphs, and adults. The results showed that the central nervous system in all the life stages of R. sanguineus consists of a mass of fused nerves named synganglion. Externally, this organ does not present segmentation, consisting of a single structure located ventromedially in the third anterior of the body. This organ is externally covered by the neural lamella or neurilemma, a uniform and acellular layer. Below, there is the periganglionic membrane or perineurium, formed by glial cells, which are characterized by their elongated nuclei. The esophagus penetrates the synganglion dividing it in two regions: supraesophageal, which is the smaller of the two and consists of a protocerebrum, a single dorsal ganglion located anteriordorsally to the esophagus; and the subesophageal, which is the largest part of the synganglion, located in the posterior region and ventral to the esophagus. Internally, the synganglion is subdivided in an external cortical region-which contains the cellular body of the nervous cells-and an internal neuropile formed by a set of neural fibers and it is constituted by bilaterally symmetric ganglia, from where nerves emerge towards several parts of the body. The results here obtained showed that there are no differences in the morpho-histology of this tissue in different life stages of R. sanguineus, suggesting that the initial structure is maintained during the whole life cycle of the tick, i.e., from larval to adult stage.
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http://dx.doi.org/10.1007/s00436-011-2785-6DOI Listing
June 2012

Morphological analysis of colon goblet cells and submucosa in type I diabetic rats submitted to physical training.

Microsc Res Tech 2012 Jun 30;75(6):821-8. Epub 2011 Dec 30.

Department of Biology, São Paulo State University (UNESP), Rio Claro, SP, Brazil.

Colon layers, especially the submucosa, as well as the secretion of goblet cells are extremely important for the functioning and transit of substances in this organ. However, the damages arising from type I diabetes and the effects of physical training, which plays crucial role in the treatment of this disease, are not yet known in these regions. To analyze the changes in colon submucosa and goblet cells of diabetic rats, as well as the effects of physical training, Wistar rats were divided into four groups: sedentary control, trained control, sedentary diabetic (SD), and trained diabetic (TD). The training protocol consisted of swimming for 60 min a day, 5 days per week, during 8 weeks. Colon samples were collected, processed, and evaluated by histochemical and ultrastructural techniques. Goblet and submucosa cells did not show alterations in shape, size, protein and carbohydrate content, in all treatment groups. Decreased amount of collagen fibers, however, was observed in the submucosa and lamina propria of SD rats, but this alteration was recovered in TDs. The ultrastructural analysis, in turn, revealed greater quantity of Golgi apparatus cisterns in SDs, distinctly than TDs, which showed improvement in this diabetic condition. Thus, physical training was responsible for the recovery of some important diabetic alterations, possibly improving the motility of substances in the large intestine. Nevertheless, it cannot be considered alone in the treatment of this disease, requiring the combined practice of other methods.
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http://dx.doi.org/10.1002/jemt.22000DOI Listing
June 2012
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