Análise da diversidade da microbiota intestinal de ratos submetidos à ressecção da valva ileocecal e criação de esfíncter artificial

OBJETIVO: analisar através de biologia molecular a diversidade da microbiota da junção ileocecal antes e após a ressecção da válvula ileocecal e reconstrução do trânsito com e sem a criação de "neoesfíncter". MÉTODOS: Os animais foram distribuídos em dois grupos: Grupo A (n=7) com ressecção da válvula ileocecal e anastomose ileocólica término-terminal em plano único, e Grupo B (n=7) com ressecção da válvula ileocecal e anastomose ileocólica término-terminal em plano único e confecção do esfíncter artificial. Reoperados com 20 dias coletou-se novamente conteúdo intraluminar do íleo e do cólon. Das amostras coletadas, extraiu-se DNA para reação de PCR-DGGE. Os padrões de bandas eletroforéticas , gerados na reação, foram submetidos ao programa Bionumerics para análise da similaridade e da diversidade da microbiota. RESULTADOS: a diversidade da microbiota foi maior e em mais amostras do íleo do que as do cólon. O grupo com a válvula apresentou os maiores valores e variações no cólon de 2,11 a 2,93. Em três animais de cada grupo estabeleceu-se comparação da similaridade e não se assemelharam ao controle. CONCLUSÃO: a ressecção da válvula ileocecal levou à mudanças da microbiota ileal e, com a criação de novo esfíncter, as variações foram maiores.

Pré-condicionamento isquêmico em diferentes tempos e seu efeito na translocação bacteriana induzida por isquemia e reperfusão intestinal em ratos

OBJETIVO: avaliar os efeitos de diferentes tempos de pré-condicionamento isquêmico(PCI) intestinal sobre a translocação bacteriana (TB). MÉTODOS: Trinta ratos Wistar pesando 280±27g foram alocados em cinco grupos. No grupo IR (n=6), foi realizada laparotomia e a artéria mesentérica superior foi ocluída por microclampe atraumático por 30 minutos. Nos quatro grupos com pré-condicionamento (n=6 cada), antes dos 30 minutos de isquemia-reperfusão (I/R) os ratos foram submetidos a PCI de dois, cinco, dez e 15 minutos e, em seguida, ao mesmo tempo de reperfusão. Vinte e quatro horas após, para avaliar se os tempos de pré-condicionamento influenciam o aparecimento de translocação bacteriana, amostras de linfonodos mesentéricos, fígado e baço foram coletadas em condições estéreis, para quantificação de unidades formadoras de colônias bacterianas por grama de tecido (UFC/g). Sangue foi coletado para dosagem de citocinas. RESULTADOS: No grupo I/R, o total de UFC/g em linfonodos mesentéricos, baço, fígado, bem como, a dosagem sérica de TNF-a, IL-1b e IL-6 foram significativamente maiores do que nos demais grupos (p<0,05). Pré-condicionamento de 15 minutos atenuou significativamente a BT e as citocinas séricas, comparando com os outros tempos de pré-condicionamento (p<0,05). CONCLUSÃO: Nossos dados sugerem o pré-condicionamento como fator-chave para reduzir translocação bacteriana em I/R intestinal. Numa escala de dois a 15 minutos, o melhor tempo de pré-condicionamento isquêmico para a atenuação da translocação bacteriana foi 15 minutos.

Modelo experimental para restrição do crescimento fetal em ratos: efeito sobre o glicogênio hepático e morfometria intestinal e renal

OBJETIVO: avaliar a eficácia do modelo de RCIU por ligadura da artéria uterina simulando insuficiência placentária em ratos. MÉTODOS: fetos de ratas prenhes Sprague-Dawley foram divididos em três grupos: RCIU (restrição de crescimento intrauterino), com fetos submetidos à ligadura da artéria uterina com 18,5 dias de gestação (termo = 22 dias), C-RCIU (controle da restrição), com fetos do corno contralateral à ligadura, CE (Controle Externo), com fetos de ratas sem manipulação. Com 21,5 dias de gestação, foi realizada cesárea, os fetos foram pesados e dissecados para análise morfométrica e histológica do fígado, intestino e rins. RESULTADOS: os dados morfométricos avaliados mostraram o peso corpóreo (PC), hepático (PH) e intestinal (PI) dos fetos com RCIU menor que C-RCIU e CE (p<0,001). O peso placentário (PP), renal (PR) e as relações PH/PC, PI/PC e PR/PC não se alteraram. A espessura renal foi menor nos fetos com RCIU (p<0,001) e houve diminuição da camada mucosa e submucosa intestinal (p<0,05). A avaliação histológica mostrou diminuição do glicogênio hepático nos fetos com RCIU em relação aos grupos C-RCIU e CE. CONCLUSÕES: o modelo descrito foi eficiente e causou RCIU fetal simétrica com diminuição da maioria dos órgãos, especialmente do peso hepático, e alteração nos depósitos de glicogênio.

Doença inflamatória intestinal - Doença de Crohn e gravidez: relato de caso

A Doença de Crohn, uma forma de doença inflamatória intestinal, é frequente em mulheres em idade reprodutiva. Seu acompanhamento requer maior atenção durante a gravidez. Apresentamos caso de doença de Crohn refratária em paciente grávida que evoluiu com ileocolectomia no puerpério. Foi revista a literatura sobre doença de Crohn na gravidez, incluindo aconselhamento das pacientes e investigação de doença ativa, além de sintetizados os dados existentes sobre a segurança dos medicamentos usados para tratá-la durante a gravidez e amamentação.

Hofbauer cells morphology and density in placentas from normal and pathological gestations

PURPOSE: In placentas from uncomplicated pregnancies, Hofbauer cells either disappear or become scanty after the fourth to fifth month of gestation. Immunohistochemistry though, reveals that a high percentage of stromal cells belong to Hofbauer cells. The aim of this study was to investigate the changes in morphology and density of Hofbauer cells in placentas from normal and pathological pregnancies. METHODS: Seventy placentas were examined: 16 specimens from normal term pregnancies, 10 from first trimester's miscarriages, 26 from cases diagnosed with chromosomal abnormality of the fetus, and placental tissue specimens complicated with intrauterine growth restriction (eight) or gestational diabetes mellitus (10). A histological study of hematoxylin-eosin (HE) sections was performed and immunohistochemical study was performed using the markers: CD 68, Lysozyme, A1 Antichymotrypsine, CK-7, vimentin, and Ki-67. RESULTS: In normal term pregnancies, HE study revealed Hofbauer cells in 37.5% of cases while immunohistochemistry revealed in 87.5% of cases. In first trimester's miscarriages and in cases with prenatal diagnosis of fetal chromosomal abnormalities, both basic and immunohistochemical study were positive for Hofbauer cells. In pregnancies complicated with intrauterine growth restriction or gestational diabetes mellitus, a positive immunoreaction was observed in 100 and 70% of cases, respectively. CONCLUSIONS: Hofbauer cells are present in placental villi during pregnancy, but with progressively reducing density. The most specific marker for their detection seems to be A1 Antichymotrypsine. It is remarkable that no mitotic activity of Hofbauer cells was noticed in our study, as the marker of cellular multiplication Ki-67 was negative in all examined specimens.

Relationship of intestinal histology with the resistance to Trichostrongylus colubriformis infection in three breeds of sheep

The study was carried out to evaluate the relationship of inflammatory intestinal cells with the resistance to Trichostrongylus colubriformis infections in three breeds of sheep (Santa Ines, Suffolk and Ile de France), naturally infected. Mast cells, eosinophils, and globule leucocytes were enumerated in intestinal mucosa. Histamine concentration was estimated in intestinal tissue samples and the length of male and female specimens were determined. The three breeds of sheep showed similar cellular response in the small intestine mucosa (P>0.05). There was extensive variation among sheep in the parasitological and inflammatory cell variables, even in lambs of the same breed. In general, animals presenting less inflammatory cells had a larger worm burden, higher fecal egg counts, and larger T. colubriformis worms. The inflammatory cells possibly impaired the parasite's establishment, development, and survival.

Intestinal lesions in pigs affected with postweaning multisystemic wasting syndrome

Samples of mesenteric lymph nodes and intestines from 79 unthrifty 3- to 5-month-old postweaning pigs, confirmed as naturally affected with postweaning multisystemic wasting syndrome (PMWS), were studied. Pigs originated from 12 farms in southern Brazil and were selected on the basis of clinical signs and/or gross lesions suggestive of enteric disorder. Lymphohistiocytic infiltrates of varying intensity were associated with anti-porcine circovirus type 2 (anti-PCV2) immunostaining (IS) in samples of intestines and mesenteric lymph nodes from all pigs. Although most findings were similar to those described in PCV2-associated enteritis, anti-PCV2 IS in association with depletion of the goblet cell mucin stores (24 pigs), diffuse ileal villous atrophy and fusion (18 pigs), and dilatation of the lymphatic vessels (11 pigs) combined or not with lymphangitis were also observed. PCV2 antigen was immunohistochemically demonstrated in the cytoplasm and nuclei from intralesional epithelial cells, histiocytes, and endothelial-like cells in intestinal tissues. Together these findings imply an association with PCV2. The presence of co-infections by Lawsonia intracellularis, Brachyspira spp., Mycobacterium spp., Salmonella spp., rotavirus, parvovirus, coronavirus and enteric calicivirus with PCV2 in the intestinal lesions was investigated.

Proliferative enteropathy (Lawsonia intracellularis) outbreak in rabbits in Brazil

An outbreak of Lawsonia intracellularis infection in rabbits, which occurred in 1988 in Rio de Janeiro state, Brazil, is reported. The disease had an acute course (24-48 hours) with clinical signs characterized by brownish or green diarrhea and dehydration. Occasionally, the animals died one day after the onset of diarrhea, without showing any other clinical signs. At necropsy, the ileum was prominent, firm and had a thickened wall; it was dilated in the caudal direction and had a somewhat reticulated appearance, perceptible through the serosa. The thickened mucous membrane had finely corrugated aspect and a shiny surface. The ileocecal valve and surrounding areas were slightly edematous and irregular. The Peyer's patches were sometimes more evident. There was moderate enlargement of the mesenteric lymph nodes. The histological examination revealed different degrees of hyperplasia of the epithelial cells of intestinal crypts consisting of poorly differentiated, hyperchromatic cells with high mitotic index, arranged in a pseudostratified layer which, in some cases, reached the apical portions of the villi. The inflammatory infiltrate between the hyperplastic epithelial cells was composed of lymphocytes, plasma cells, macrophages, some eosinophils and globular leukocytes. Silver impregnation revealed large numbers of bacteria with morphology of the genus Lawsonia in the apical pole of cryptal enterocytes. These bacteria reacted positively to a Lawsonia intracellularis polyclonal antibody by the avidin-biotin immunohistochemistry method.

Paratuberculosis in a dairy Gyr herd in the State of Paraíba, Brazil

This paper describes the clinical, pathological, and microbiologic aspects of paratuberculosis (Johne's disease) in a dairy Gyr herd in the State of Paraíba, northeastern Brazil. An eight years old cow with chronic unresponsive diarrhea was clinically examined and euthanized for pathological evaluation. Fecal samples from all 160 animals over 12 months of age from the herd were collected for isolation of Mycobacterium avium subsp. paratuberculosis. Clinically, the index case cow was severely dehydrated, cachectic, with profuse mucous diarrhea. The main post-mortem findings were emaciation and thickened intestinal wall. Microscopically, the intestinal lamina propria and submucosa were infiltrated by macrophages, epithelioid cells, and Langhans giant cells with numerous alcohol-acid resistant bacilli in the cytoplasm. Two fecal samples displayed growth in slants of Herrold's egg-yolk agar supplemented with mycobactin J, 150 days after incubation. No growth was noticed in slants without mycobactin J. Microscopic examination of the isolated microorganisms stained by Ziehl-Neelsen revealed considerable amounts of alcohol-acid resistant bacilli, morphologically compatible with Mycobacterium spp. Based on the clinical signs, gross and histological lesions, growth time, bacterial morphology in Ziehl-Neelsen staining, and dependence of mycobactin J, the first diagnosis of paratuberculosis in Zebu cattle was made.

Absence of intestinal colonization by vancomycin-resistant enterococci in nonhuman primates

The animal reservoirs of vancomycin-resistant enterococci (VRE) have important role in the epidemiology of the bacteria and resistant genes. The present work searched fecal samples taken off nonhuman primates for the presence of VRE. Resistance profiles, virulence traits, and genetic variability among enterococci isolates were also analyzed. The samples included Capuchin monkeys (Cebus apella, n=28) and Common marmoset (Callithrix penicillata, n=37) housed in the Primate Center of the University of Brasília, Brazil. Most individuals were captive monkeys from the Central-West and South-East regions of Brazil (n=48). We collected rectal swabs and carried out selective isolation followed by multiplex Polymerase Chain Reaction (PCR) to identify species and resistance genes. No vanA or vanB-containing enterococci were found. The carriage rates ranged from 1.5% for the VanC-type E. casseliflavus and E. gallinarum until 12.3% (n=8) for Enterococcus faecalis. All E. faecalis isolates showed susceptibility to vancomycin, teicoplanin, ampicillin, gentamicin, and streptomycin. The virulence genes ace and esp were prevalent (100.0%, 87.5%). Multilocus variable number of tandem repeats (MLVA) revealed diversity in the number of repeats among E. faecalis isolates and targets, which was higher for espC, efa5, and efa6. We identified six different MLVA genotypes that were divergent from those described in human beings. Also, they were clustered into two genogroups that showed host-specificity for the species Cebus apella or Callithrix penicillata. In conclusion, no vanA- or vanB-containing enterococci were found colonizing those primate individuals. This finding suggested that the primate individuals investigated in our study are not directly involved in the epidemiological chain of high-level vancomycin-resistant genes vanA or vanB in Brazil. Our study also showed that E. faecalis isolated from nonhuman primates carry virulence traits and have ability to spread their lineages among different individuals.

Effects of infection with Toxoplasma gondii oocysts on the intestinal wall and the myenteric plexus of chicken (Gallus gallus)

This paper aims to analyze the effects of the Toxoplasma gondii infection in the intestinal wall and myenteric plexus of chicken (Gallus gallus). Ten 36-day-old chickens were separated into two groups: control and experimental, orally inoculated with oocysts of the T. gondii strain M7741 genotype III. After 60 days the birds were submitted to euthanasia and had their duodenum removed. Part of the intestinal segments was submitted to histological routine, HE staining, PAS histochemical technique, and Alcian Blue. Qualitative analysis of the intestinal wall and comparative measurements among the groups with respect to total wall thickness, muscle tunic, mucosa, and tunica mucosa were carried out. Caliciform cells were quantified. The other part of the intestinal segments was fixed in formol acetic acid and dissected having the tunica mucosa and the tela submucosa removed. Neurons were stained with Giemsa, counted, and measured. Chickens from the experimental group presented diarrhea and inflammatory infiltrates in the tunica mucosa, thickness reduction of all the parameters assessed in the intestinal wall, and an increase of the number of caliciform cells. There was a ~70% reduction regarding the intensity of myenteric neurons; and the remaining cells presented a reduction of ~2.4% of the perikarion and ~40.5% of the nucleus (p<0.05). Chronic infection induced by T. gondii oocysts resulted in intestinal wall atrophy, mucin secretion increase, death and atrophy of chicken myenteric plexus neurons. Death and atrophy of myenteric plexus neurons may be related with the causes of diarrhea observed in chickens with toxoplasmosis.

Morphology of the male agouti accessory genital glands (Dasyprocta prymnolopha Wagler, 1831)

The morphology of the accessory genital glands of the male agouti was studied in twenty-three animals that were raised in captivity. Twenty animals had their genital glands dissected in situ for macroscopic description. The samples of each gland were recovered, embedded in paraffin, sliced and stained by Hematoxylin-Eosin technique. It was founded four pairs of glands: the vesicular glands, the coagulating glands, the prostate and the bulbourethral glands. Histological characteristics of the vesicular, coagulating and prostate glands showed similar morphology, within the pseudostratified columnar epithelium. The tubulo-alveolar type of the bulbourethral glands showed a lack of connective tissue among the tubules, a small amount of red stained presented it the cytoplasm, and the presence of vacuoles in the tissue. This study concluded that the agouti showed to have similar morphological aspect described in the others species of rodents.

Morphology and topographic anatomy of the spinal cord of the red-footed tortoise (Geochelone carbonaria Spix, 1824)

The aim of this study was to describe the topography of the spinal cord of the red-footed tortoise to establish a morphological basis for applied research in anesthesiology and morphology. Six tortoises from the state of Maranhão (Brazil) that had died of natural causes were used. The common carotid artery was used to perfuse the arterial system with saline solution (heated to 37ºC) and to fix the material with a 20% formaldehyde solution. The specimens were then placed in a modified decalcifying solution for 72 hours to allow dorsal opening of the carapace with a chisel and an orthopedic hammer. Dissection of the dorsal musculature and sectioning of the vertebral arches were performed to access the spinal cord. The results revealed the spinal cord of G. carbonaria to be an elongated, whitish mass that reached the articulation between the penultimate and last caudal vertebrae. The cervical intumescence (Intumescentia cervicalis) was located between vertebral segments C5 and T1, whereas the lumbosacral intumescence (Intumescentia lumbalis) was located between T6 and Ca1.

Saimiri sciureus' hard palate morphology

Primate order includes around 180 species. Morphological aspects of New World non-human primates (NHP) have been extensively investigated since last century. General commonsense describes oral cavity adaptations according to diet and feeding, dentition, tongue projection and head shape. Morphological appearance and dimension of the hard palate have been outstanding as interest in many species including man. Six young Saimiri sciureus hard palate were investigated. We measured the hard palate distance (HL), intercanine distance (ICD), intermolar distance (IMD), and interpremolar distance (IPD). Complete and incomplete palatine crests were quantified. We believe that better understanding of the mouth roof morphology will contribute to improve the management of captive animal's diet in order to re-introduce the animals in its habitat.

Período de absorção intestinal de macromoléculas em cabritos recém-nascidos após a ingestão de colostro bovino

Após o nascimento, os cabritos são dependentes das imunoglobulinas colostrais devido às características placentárias que não permitem a passagem de macromoléculas da circulação materna. De acordo com a literatura, os cabritos possuem capacidade absortiva por até quatro dias. Muitos aspectos fisiológicos de outras espécies são aceitos e utilizados para caprinos, mas aqueles relacionados à transferência de imunidade passiva precisam de investigação. Os objetivos do presente estudo foram determinar o período de passagem de macromoléculas da mucosa intestinal para a circulação e a duração da proteção humoral transferida passivamente pela ingestão de colostro bovino e caprino. Sessenta cabritos recém-nascidos foram distribuídos em seis tratamentos: T 0 (n=25), ingestão natural de colostro caprino à vontade; T 1 (n=7), colostro bovino entre o nascimento e duas horas pós-parto; T 2 (n=7), ingestão de colostro bovino entre quatro e seis horas pós-nascimento; T 3 (n=7), leite nas primeiras oito horas e colostro bovino entre 10 e 12 horas pós-parto; T 4 (n=7), ingestão de leite até 18 horas e colostro bovino entre 22 e 24 horas pós-nascimento; T 5 (n=7), leite até 30 horas e ingestão de colostro bovino entre 34 e 36 horas pós-parto. Determinaram-se as concentrações séricas de proteína total (PT), gamaglobulina, imunoglobulina G (IgG) e a atividade sérica de gama glutamiltransferase (GGT). Ao nascimento, todos os neonatos tiveram valores mais baixos das variáveis, com aumento significativo da PT e gamaglobulina, após dois dias, nos grupos T 0, T 1 e T 2; a IgG e GGT aumentaram em todos os grupos. Os tratamentos T 3, T 4 e T 5 foram considerados como indutores de falha de transferência de imunidade passiva. A absorção de macromoléculas pelo trato intestinal dos cabritos ocorreu até 36 horas pós-parto, sendo mais efetiva até 12 horas. Os níveis de anticorpos persistiram até 75 dias após a ingestão de colostro bovino, porém, com concentrações inadequadas.