Debora de Mello Gonçales Sant'Ana

Morphological and quantitative study of the myenteric plexus of the ascending colon of rats subjected to proteic desnutrition

We carried out this study with the purpose of contributing on the effects of the proteic desnutrition on the morphological aspects and quantitative analysis of the neurons in the myenteric plexus of the ascending colon of adult Rattus norvegicus. Twenty adult rats were divided into two groups: in one of them, we offered a normal ration with proteic level of 22% (control group) and in the other, a ration with a proteic level of 8% (experiment group) during 120 days. We did the whole-mount preparations for the ascending colon and stained them with the Giemsa technique and the histochemical technique of NADH-diaphorase. The rats with proteic desnutrition showed a body weight, on average, to be 35.1% less than those of the control group, and the colon was on average, 26.8% shorter and 6.7% narrower. Thus, it was to be expected that the colon of animals with proteic desnutrition had a neuronal density 31.62% greater than the rats of the control group. Nevertheless, the difference with the Giemsa technique was on average 18.4%, demonstrating a mean neuronal loss of 13.25%.

Effects of protein and vitamin B deficiency on blood parameters and myenteric neurons of the colon of rats

The aims of this work were to evaluate the effects of the deficient ingestion of protein and vitamin B on the biochemical and hematologic parameters and on the NADH- and NADPH-diaphorase positive myenteric neurons. The control animals (n=10) received commercial chow and the experimental rats (n=10) received chow with protein level reduced to 8% during 120 days. At the time of killing blood was collected for assessment of the blood and hematologic parameters and the ascending colon for quantitative analysis of the neurons of the myenteric plexus. It was observed that the reduction of the protein level to 8% coupled to the reduction of the levels of vitamin B in adult rats neither led to qualitative or quantitative changes on red or white blood cells, nor decreased globulin levels, induced the formation of edema or gave rise to clinical signs typical of protein or vitamin B deficiency. On the other hand, the experimental protocol led to less weight gain, change on the body composition with fat deposition; decrease of the values of serum total protein and albumin; reduction of the area of colon and density of nitrergic and NADH-diaphorase myenteric neurons inferior to the expected.

Number and size of myenteric neurons of the duodenum of adult rats with acute diabetes

This study had as its purpose to assess the effects of acute diabetes induced by streptozotocin (35 mg/kg body weight) on the number and size of the myenteric neurons of the duodenum of adult rats considering equally the antimesenteric and intermediate regions of the intestinal circumference. Experimental period extended for a week. Neuronal counts were carried out on the same number of fields of both regions of the duodenal circumference and measurements of neuronal and nuclear areas on equal numbers of cells. Number and size of the myenteric neurons stained with Giemsa were not significantly different between groups. On the other hand, the proportion of NADH-positive neurons increased from 18.54% on the controls to 39.33% on the diabetics. The authors discuss that this increased reactivity probably results from a greater NADH/NAD+ ratio, described in many tissues of diabetic animals, which has consequences on the modulation of the enzymes that use these cofactors and whose activity is detected by the NADH-diaphorase technique.

Alterations of the myenteric plexus of the ileum and the descending colon caused by Toxoplasma gondii (genotype III)

Alterations caused by a genotype III strain of Toxoplasma gondii were assessed with respect to the number and the morphometry of the myenteric neurons in the terminal ileum and the descending colon. Eighteen rats were divided into four groups: Acute Control Group (ACG, n=4); Acute Experimental Group (AEG, n=4); Chronic Control Group (CCG, n=5) and Chronic Experimental Group (CEG, n=5). NaCl solution was administered through gavage to the animals in the ACG and CCG. Toxoplasma gondii tachyzoites (10(4)) from a genotype III strain were orally administered to the AEG and CEG. Acute Groups were died after 24 hours, and the Chronic Groups after 30 days. Neuronal loss was not observed in both organs. The neurons atrophied in the terminal ileum as the opposite occurred with the neurons at the descending colon during the chronic phase of infection. In the terminal ileum, the neurons atrophied during the chronic phase of the infection as no alteration was found during the acute phase. For the descending colon, the neurons became hypertrophic during the chronic infection in opposition to the atrophy found during the acute phase.

Alterações do epitélio branquial e das lamelas de tilápias (Oreochromis niloticus) causadas por mudanças do ambiente aquático em tanques de cultivo intensivo

The behavior of the gill epithelium of tilapias cultured in tanks at different altitudes and interconnected with PVC pipes was analyzed. Gill filaments of four specimens from four tanks (T1, T2, T3 e T4) sequentially interconnected were submitted to histological routine to obtain 5-mm-thick cuts that were stained with HE or submitted to histochemistry reactions PAS + diastase solution or Alcian Blue pH 2.5 or Alcian Blue pH 1.0. Considering the intermediary, apical and basal regions of the filaments, the lamellar area was measured and the amount of mucous cells was counted. It was verified that oxygen, pH, and temperature decreased progressively as the water flew from one tank to another. Thus, an increase was realized of the amount of mucous cells and the lamellar area in T2, as well as a progressive decrease of these measures on the tanks which received water from T2. Moreover, detachment of the gill epithelium, cellular hyperplasia in the interlamellar space, and telangectasias were observed in the fishes from T2, T3 e T4. It was concluded that the aquatic environment in tanks sequentially interconnected with PVC pipes suffers alterations from one tank to another, as that physico-chemical fluctuations reflect on the behavior of the gill epithelium through variations of the lamellar area and the amount of mucous cells.

Neuron number in the myenteric plexus of the ascending colon of rats: a comparative study using two staining techniques

We carried out this study with the purpose of comparing the neuronal density in antimesocolic and intermediate regions of the colon of rats. We used the ascending colon of ten seven-months old Wistar rats. With the Giemsa method we found 29,046 neurons/cm2 on the antimesocolic region and 30,968 neurons/cm2 on the intermediate regions. With the NADH-diaphorase technique 12,308 neurons/cm2 on the antimesocolic regions and 8798 neurons/cm2 on the intermediate regions were evidenced. The number of NADH-diaphorase positive neurons is significantly less than the number of Giemsa-stained neurons and that this difference is enhanced on the intermediate regions of the intestinal circumference. Therefore, to compare the number of neurons of an intestinal segment of a same species at the same age, it is necessary to take into consideration the technique employed and the region of the intestinal circumference from where the sample was obtained.

Quantitative analysis of the neurons from the myenteric plexus in the ileum of rats submitted to severe protein deficiency

The effects of protein malnutrition on the quantitative aspects of the myenteric plexus in the ileum of adult Rattus norvegicus were assessed. Thirty 90-day-old rats were divided into two groups: Control Group (CG, n=15) and Experimental Group (EG, n=15). The CG received 26% protein chow and the EG received 4% protein chow for 90 days. At the end of the experiment, the animals from the CG weighed 369.63+/-26.33, and the ones from the EG 215.34+/-56.31. The ileum was submitted to Giemsa, NADH- and NADPH-diaphorase technique in order to evidence nervous cells in the whole-mount preparations. Animals from the EG presented a 41.75% body weight loss in relation to the CG as well as 17.6% length reduction for the ileum-jejunum. Moreover, the organ was 41% lighter for the EG. Giemsa-stained neurons were 17.02% more concentrated in the EG (p>0.05). NADH-diaphorase-stained neurons were 26.6% more concentrated in the EG (p<0.05), while the NADPH-diaphorase were 26.28% more concentrated in this group (p<0.05).

Probiotics prevent growth deficit of colon wall strata of malnourished rats post-lactation

The objective of this study was to analyze morphometrically the colon wall strata of malnourished rats supplemented with probiotics. Sixteen recently weaned Wistar rats (Rattus norvegicus) were distributed into four groups: animals that received commercial chow (G1, n = 4); animals that received the same feed as G1 and were supplemented with probiotics (G2, n = 4); animals that received chow with 4% of proteins (G3, n = 4); animals that received the same feed as G3 and were supplemented with probiotics (G4, n = 4). After 12 weeks, the proximal colon was collected and submitted to histological processing. Three-µm cuts were stained with H.E., Periodic Acid Schifff (P.A.S.) + diasthasis solution and Alcian Blue (A.B.) pH 2.5 and pH 1.0. The morphometric analysis of the intestinal wall showed that the supplementation with ABT-4 probiotic culture prevents the growth deficit of colon wall strata that normally occurs in malnourished rats right after lactation. Besides, no alteration was observed in the proportion of the number of globet cells in relation to the number of enterocytes in malnourished rats, regardless of the supplementation with probiotics.

Neurônios NADH-diaforase positivos do jejuno de ratos adultos (Rattus norvegicus) desnutridos: aspectos quantitativos

We aim at contributing with information on the quantitative aspects of the NADH-diaphorase positive myenteric neurons of the jejunum of adult rats subjected to protein desnutrition. Ten rats aging 90 days were divided into two groups: control (n=5, ±278 g) and disnurtured (n=5, ±280 g). In the following 120 days, the rats from the control group had chow with 22% protein level, and those from the disnurtured group, with 8% protein level. After this period, the control rats weighted ±394.4g and the disnurtured ±273.5g. The jejunum was subjected to the histochemical technique of the NADH-diaphorase to stain nerve cells in whole-mounts. The neurons found in 80 microscopic fields of both groups were counted. In the control ±674.6 neurons were observed, and ±1326.8 neurons were counted in the disnurtured group. The low-protein diet did not alter the organization of the neurons, but led to a smaller body growth in the disnurtured animals, preventing neuronal dispersal and leading to a greater density per mm2.

Oral dependent-dose toxoplasmic infection model induced by oocysts in rats: Myenteric plexus and jejunal wall changes

Toxoplasmosis is a widely distributed disease caused by the protozoan Toxoplasma gondii that is mainly transmitted orally. Once ingested, the parasite crosses the intestinal barrier to reach the blood and lymph systems to migrate to other regions of the host. The objective of this study was to evaluate the changes in the myenteric plexus and the jejunal wall of Wistar rats caused by oral infection with T. gondii oocysts (ME-49 strain). Inocula of 10, 100, 500 and 5000 oocysts were used. The total population of myenteric neurons and the most metabolically active subpopulation (NADH-diaphorase positive - NADH-dp) exhibited a decrease proportional to the dose of T. gondii. There was also a quantitative increase in the subpopulation of NADPH-diaphorase-positive (NADPH-dp) myenteric neurons, indicating greater expression of the NOS enzyme. Neuronal atrophy was observed, and morphological and morphometric alterations such as jejunal atrophy were found in the infected groups. Hypertrophy of the external muscle with the presence of inflammatory foci was observed in the group infected with 5000 oocysts. The changes observed in the infected groups were proportional to the number of oocysts inoculated.