Regional differences in the ultrastructure of mucosal macrophages in the rat large intestine.

We previously clarified the histological characteristics of macrophages in the rat small intestine using serial block-face scanning electron microscopy (SBF-SEM). However, the regional differences in the characteristics of macrophages throughout the large intestine remain unknown. Here, we performed a pilot study to explore the regional differences in the ultrastructure of mucosal macrophages in the large intestine by using SBF-SEM analysis. SBF-SEM analysis conducted on the luminal side of the cecum and descending colon revealed macrophages as amorphous cells possessing abundant lysosomes and vacuoles. Macrophages in the cecum exhibited a higher abundance of lysosomes and a lower abundance of vacuoles than those in the descending colon. Macrophages with many intraepithelial cellular processes were observed beneath the intestinal superficial epithelium in the descending colon. Moreover, macrophages in contact with nerve fibers were more prevalent in the cecum than in the descending colon, and a subset of them surrounded a nerve bundle only in the cecum. In conclusion, the present pilot study suggested that the quantity of some organelles (lysosomes and vacuoles) in macrophages differed between the cecum and the descending colon and that there were some region-specific subsets of macrophages like nerve-associated macrophages in the cecum.

Zearalenone Exposure Triggered Cecal Physical Barrier Injury through the TGF-ß1/Smads Signaling Pathway in Weaned Piglets.

This study aims to investigate the effects of exposure to different dosages of zearalenone (ZEA) on cecal physical barrier functions and its mechanisms based on the TGF-ß1/Smads signaling pathway in weaned piglets. Thirty-two weaned piglets were allotted to four groups and fed a basal diet supplemented with ZEA at 0, 0.15, 1.5, and 3.0 mg/kg, respectively. The results showed that 1.5 and 3.0 mg/kg ZEA damaged cecum morphology and microvilli, and changed distribution and shape of M cells. Moreover, 1.5 and 3.0 mg/kg ZEA decreased numbers of goblet cells, the expressions of TFF3 and tight junction proteins, and inhibited the TGF-ß1/Smads signaling pathway. Interestingly, the 0.15 mg/kg ZEA had no significant effect on cecal physical barrier functions but decreased the expressions of Smad3, p-Smad3 and Smad7. Our study suggests that high-dose ZEA exposure impairs cecal physical barrier functions through inhibiting the TGF-ß1/Smads signaling pathway, but low-dose ZEA had no significant effect on cecum morphology and integrity through inhibiting the expression of smad7. These findings provide a scientific basis for helping people explore how to reduce the toxicity of ZEA in feeds.

Examination of intestinal ultrastructure, bowel wall apoptosis and tight junctions in the early phase of sepsis.

Gut hyperpermeability can be caused by either apoptosis of the intestinal epithelium or altered status, permeability or porosity of tight junctions. This project aims to elucidate these mechanisms in the early phase of sepsis. Eighteen male wild type mice were randomized to two groups. All mice received one single gavage of fluorescein isothiocyanate (FITC) dextran 30 min before intervention. One group (n = 10) underwent cecal ligation and puncture to induce sepsis. The other group (n = 8) was sham operated. Septic animals exhibited significantly increased permeability for FITC 8 h post-operatively. Significantly increased serum interleukin-6, tumor-necrosis-factor-alpha and interleukin-1-beta confirmed sepsis. Septic animals showed significant bowel wall inflammation of ileum and colon samples. PCR revealed significantly increased expression of claudin-2 and decreased expressions of claudin-4, tight-junction-protein-1 and occludin-1 resembling increased permeability of tight junctions. However, these alterations could not be confirmed at the protein level. Light microscopy revealed significant dilatation of intercellular spaces at the basal sections of intestinal epithelial cells (IEC) in septic animals confirmed by increased intercellular spaces at the level of tight junctions and adherens junctions in electron microscopy (TEM). In small angle X-ray scattering no increase in number or size of nanopores could be shown in the bowel wall. HOECHST staining and PCR of ileum samples for apoptosis markers proofed no relevant differences in intestinal epithelial cell apoptosis between the groups. Intestinal hyperpermeability in septic animals was most likely caused by alterations of the intercellular contacts and not by apoptosis or increased size/number of nanopores of intestinal epithelial cells in this murine model of early sepsis.

Paternal weight of ducks may have an influence on offspring' small intestinal function and cecal microorganisms.

BACKGROUND: In animals, many factors affect the small intestinal function and cecal microorganisms, including body weight and genetic background. However, whether paternal weight impacts the small intestinal function and cecal microorganisms remains unknown to date. The current study used Nonghua sheldrake to estimate the effect of paternal weight on the intestine of the offspring by evaluating differences in small intestinal morphology, digestive enzyme activity, and cecal microorganisms between the offspring of male parents with high body weight (group H) and that of male parents with low body weight (group L). RESULTS: The results of the analysis of small intestinal morphology showed that the villus height of the jejunum of group H ducks was higher than that of group L ducks, and the difference was significant for ducks at 10 weeks of age. Moreover, the villus height/crypt depth of the duodenum in group H significantly exceeded that of group L at a duck age of 2 weeks. The amylase activity in the jejunum content of group H exceeded that of group L at 5 and 10 weeks of age. Furthermore, the proportion of the Firmicutes to Bacteroidetes was significantly higher in group H (duck age of 2 weeks). Among the genera with a relative abundance exceeding 1%, the relative abundances of genera Desulfovibrio, Megamonas, Alistipes, Faecalibacterium, and Streptococcus observed in group H were significantly different between group H and group L. CONCLUSIONS: For the first time, this study identifies the effect of paternal weight on offspring small intestinal function and cecal microorganisms. Consequently, this lays a foundation for further research on the relationship between male parents and offspring intestinal function.

Cellular elements in the developing caecum of Japanese quail (Coturnix coturnix japonica): morphological, morphometrical, immunohistochemical and electron-microscopic studies.

The present study aims to investigate the histological, histochemical and electron microscopic changes of the caecal proximal part of Japanese quail during both pre- and post-hatching periods starting from the 2nd embryonic day (ED) until four weeks post-hatching. On the 2nd and 3rd ED, the primordia of caeca appeared as bilateral swelling on the wall of the hindgut. On the 7th ED, the lamina propria/submucosa contained the primordia of glands. On the 8th ED, rodlet cells could be observed amongst the epithelial cells. On the 9th ED, the caeca began to divide into three parts with more developed layers. With age, the height and number of villi increased. On the 13th ED, immature microfold cells (M-cells) could be identified between the surface epithelium of the villi. The caecal tonsils (CTs) appeared in the form of aggregations of lymphocytes, macrophages, dendritic cells and different types of leukocytes. Telocytes and crypts of Lieberkuhn were observed at this age. On hatching day, the crypts of Lieberkuhn were well-defined and formed of low columnar epithelium, goblet cells, and enteroendocrine cells. Post-hatching, the lumen was filled with villi that exhibited two forms: (1) tongue-shaped villi with tonsils and (2) finger-shaped ones without tonsils. The villi lining epithelium contained simple columnar cells with microvilli that were dispersed with many goblet cells, in addition to the presence of a high number of intra-epithelial lymphocytes and basophils. Moreover, the submucosa was infiltrated by numerous immune cells. CD3 immunomarker was expressed in intraepithelial lymphocytes, while CD20 immunomarker showed focal positivity in CTs. In conclusion, the caecal immune structures of quails at post-hatching were more developed than those in pre-hatching life. The high frequency of immune cells suggests that this proximal part may be a site for immunological surveillance in the quail caecum. The cellular organisation of the caecum and its relation to the immunity was discussed.

Mapping the intermediate digestion phases of human healthy intestinal contents from distal ileum and caecum at fasted and fed state conditions.

OBJECTIVES: To investigate at the ultrastructural level, the colloidal phases formed in the lumen of the distal ileum and caecum of healthy adults. METHODS: Cryogenic transmission electron microscopy (Cryo-TEM) was employed to image the intermediate colloidal phases of human intestinal contents collected from distal ileum and caecum of two healthy volunteers under fasted and fed state conditions. KEY FINDINGS: In samples collected both in the fasted and fed states, Cryo-TEM study revealed the presence of large spherical unilamellar and occasionally bi-lamellar and oligolamellar vesicles with diameters ranging from 50 to 200 nm for both volunteers in distal ileum and caecum. Bilayer fragments were frequently observed in caecal samples. Plate-like structures resembling the morphology of cholesterol plates were visualised in all samples. Elongated structures were observed in the fed state in distal ileum and caecum for both volunteers, whereas no micellar structures could be detected for all samples. CONCLUSIONS: This study provides a framework for understanding the structure of colloidal phases, and it may assist in elucidating the role of dosing conditions on drug absorption from the distal ileum and caecum.

Critical role of microbiota within cecal crypts on the regenerative capacity of the intestinal epithelium following surgical stress.

Cecal crypts represent a unique niche that are normally occupied by the commensal microbiota. Due to their density and close proximity to stem cells, microbiota within cecal crypts may modulate epithelial regeneration. Here we demonstrate that surgical stress, a process that invariably involves a short period of starvation, antibiotic exposure, and tissue injury, results in cecal crypt evacuation of their microbiota. Crypts devoid of their microbiota display pathophysiological features characterized by abnormal stem cell activation as judged by leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) staining, expansion of the proliferative zone toward the tips of the crypts, and an increase in apoptosis. In addition, crypts devoid of their microbiota display loss of their regenerative capacity as assessed by their ability to form organoids ex vivo. When a four-member human pathogen community isolated from the stool of a critically ill patient is introduced into the cecum of mice with empty crypts, crypts become occupied by the pathogens and further disruption of crypt homeostasis is observed. Fecal microbiota transplantation restores the cecal crypts' microbiota, normalizes homeostasis within crypts, and reestablishes crypt regenerative capacity. Taken together, these findings define an emerging role for the microbiota within cecal crypts to maintain epithelial cell homeostasis in a manner that may enhance recovery in response to the physiological stress imposed by the process of surgery. NEW & NOTEWORTHY: This study provides novel insight into the process by which surgical injury places the intestinal epithelium at risk for colonization by pathogenic microbes and impairment of its regenerative capacity via loss of its microbiota. We show that fecal transplant restores crypt homeostasis in association with repopulation of the microbiota within cecal crypts.

Molecular Characterization and Immune Protection of a New Conserved Hypothetical Protein of Eimeria tenella.

The genome sequences of Eimeria tenella have been sequenced, but >70% of these genes are currently categorized as having an unknown function or annotated as conserved hypothetical proteins, and few of them have been studied. In the present study, a conserved hypothetical protein gene of E. tenella, designated EtCHP559, was cloned using rapid amplification of cDNA 5'-ends (5'RACE) based on the expressed sequence tag (EST). The 1746-bp full-length cDNA of EtCHP559 contained a 1224-bp open reading frame (ORF) that encoded a 407-amino acid polypeptide with the predicted molecular weight of 46.04 kDa. Real-time quantitative PCR analysis revealed that EtCHP559 was expressed at higher levels in sporozoites than in the other developmental stages (unsporulated oocysts, sporulated oocysts and second generation merozoites). The ORF was inserted into pCold-TF to produce recombinant EtCHP559. Using western blotting, the recombinant protein was successfully recognized by rabbit serum against E. tenella sporozoites. Immunolocalization by using EtCHP559 antibody showed that EtCHP559 was mainly distributed on the parasite surface in free sporozoites and became concentrated in the anterior region after sporozoites were incubated in complete medium. The EtCHP559 became uniformly dispersed in immature and mature schizonts. Inhibition of EtCHP559 function using anti-rEtCHP559 polyclonal antibody reduced the ability of E. tenella sporozoites to invade host cells by >70%. Animal challenge experiments demonstrated that the recombinant EtCHP559 significantly increased the average body weight gain, reduced the oocyst outputs, alleviated cecal lesions of the infected chickens, and resulted in anticoccidial index >160 against E. tenella. These results suggest that EtCHP559 plays an important role in sporozoite invasion and could be an effective candidate for the development of a new vaccine against E. tenella.

The several elements of intestinal innate immune system at the beginning of the life of broiler chicks.

Functional capacity of digestive system and intestinal adaptive immunity are immature at hatching of broiler chicks. Therefore, intestinal innate immunity after hatching is vital to young chicks. The purpose of this study was to investigate expression and tissue distributions of several elements of the innate immune system (i.e., TLR2, TLR4, CD83, and MHC class II expressing cells) in the intestine of one-day-old chicks. For this purpose, ileum and cecum were examined the under different conditions, which included the control and 1, 3, 6, 12, or 24 h after injection of lipopolysaccharide (LPS) and phosphate buffered saline. The findings indicated that regardless of the antigenic stimulation, Toll-like receptor (TLR) 2 and TLR4 expressing cells were present in the intestinal tissues of one-day-old chicks. We noticed that the intestinal segments have different TLR expression levels after LPS stimulation. Dendritic cells were identified, and they left the intestinal tissue after LPS treatment. MHC class II molecules were diffusely present in both the ileum and cecum. This study demonstrates that the intestinal tissue of one-day-old chicks has remarkable defensive material, including histological properties and several elements of the innate immune system. Microsc. Res. Tech. 79:604-614, 2016. © 2016 Wiley Periodicals, Inc.

Ultrastructural effects of acetamizuril on endogenous phases of Eimeria tenella.

To explore the primary stage or site of action of acetamizuril (AZL), a novel triazine anticoccidial compound, the ultrastructural development of Eimeria tenella at different endogenous stages was studied in experimentally infected chickens treated with a single oral dose of 15 mg/kg AZL. As a result of drug action, the differentiations of second-generation schizonts and microgamonts were largely inhibited and merozoites became irregular in shape. Meanwhile, the outer membrane blistering and perinuclear space enlargement were obvious in the second-generation schizonts and microgamonts, which were never observed in the classic triazine anticoccidiosis drug diclazuril-treated E. tenella. The chromatin aggregation, anachromasis, and marginalization were visible in merozoites and microgamonts. Furthermore, the abnormal evagination of microgametes finally resulted in the degeneration of microgamonts and the failure of subsequent fertilization. The most marked micromorphological alteration occurring in the macrogamonts was the WFB2. Even if the fertilization occurred, the formation of oocyst wall became malformed and the zygote proceeded to the obvious degeneration. In addition, mitochondria swelling and cytoplasm vacuolization were discerned in respective intracellular stages, while endoplasmic reticulum and Golgi body swelling was less seen. These alterations may be the causes leading to the final death of E. tenella and also provide some information for further exploring the mechanism of action of AZL at the molecular level.

Ultrastructural study on the morphological changes in indigenous bacteria of mucous layer and chyme throughout the rat intestine.

Indigenous bacteria in the alimentary tract are exposed to various bactericidal peptides and digestive enzymes, but the viability status and morphological changes of indigenous bacteria are unclear. Therefore, the present study aimed to ultrastructurally clarify the degeneration and viability status of indigenous bacteria in the rat intestine. The majority of indigenous bacteria in the ileal mucous layer possessed intact cytoplasm, but the cytoplasm of a few bacteria contained vacuoles. The vacuoles were more frequently found in bacteria of ileal chyme than in those of ileal mucous layer and were found in a large majority of bacteria in both the mucous layer and chyme throughout the large intestine. In the dividing bacteria of the mucous layer and chyme throughout the intestine, the ratio of area occupied by vacuoles was almost always less than 10%. Lysis or detachment of the cell wall in the indigenous bacteria was more frequently found in the large intestine than in the ileum, whereas bacterial remnants, such as cell walls, were distributed almost evenly throughout the intestine. In an experimental control of long-time-cultured Staphylococcus epidermidis on agar, similar vacuoles were also found, but cell-wall degeneration was never observed. From these findings, indigenous bacteria in the mucous layer were ultrastructurally confirmed to be the source of indigenous bacteria in the chyme. Furthermore, the results suggested that indigenous bacteria were more severely degenerated toward the large intestine and were probably degraded in the intestine.

Ultrastructural characteristics of the caeca of basal polyopisthocotylean monogeneans of the families Chimaericolidae and Hexabothriidae parasitic on cartilaginous fishes.

Ultrastructural differences are shown between the caecal organization in three blood-feeding polyopisthocotylean monogeneans, i.e., the chimaericolid Chimaericola leptogaster from the holocephalan Chimaera monstrosa and two hexabothriids, Callorhynchocotyle callorhynchi from the holocephalan Callorhynchus capensis and Rajonchocotyle emarginata from the elasmobranch Amblyraja radiata. In C. leptogaster, digestive cells and connecting syncytium, joined close to the luminal surface by septate junctions, are arranged alternately along the caecal epithelial wall; the nuclear regions of both cell types are sunk below the general level of the caecal epithelium; a concave depression on the apical margin of the digestive cells bears lamellae; and this depression is covered by a lamellate bubble formed by thin projections emanating from the connecting syncytium. The luminal surface of the connecting syncytium is covered with outgrowths terminating in the form of long, narrow processes. In R. emarginata and C. callorhynchi, the predominant digestive cells are at different stages of development and occur in groups, developing digestive cells bulge into the caecal lumen from the connecting syncytium with contact sites present close to the luminal surface, and the luminal surface structures of both the connecting syncytium and the digestive cells are short lamellae. In these two hexabothriids, a holocrine (or apocrine) process for the elimination of digestive product is assumed via the detachment of fully differentiated, bulging digestive cells. Free, apparently sloughed digestive cells and residual bodies are present within the caecal lumen, and replacement digestive cells are numerous in the connecting syncytium. In the chimaericolid, free bubbles containing residual bodies and portions of digestive cells filled with degenerating digestive vesicles occur in the caecal lumen along with large amounts of male and female reproductive material. The usefulness of characteristics of the caecal ultrastructure as taxonomic traits at the family level is discussed.

Saccharomyces boulardii prevention of the hepatic injury induced by Salmonella Enteritidis infection.

Salmonella enterica subsp. enterica serovar Enteritidis (Salmonella Enteritidis) is the predominant cause of serovar-associated food-borne outbreaks in many countries and causes significant clinical symptoms of liver injury, enteritis, and diarrheal diseases. Saccharomyces boulardii is used in clinical application for prophylaxis and the treatment of a variety of diseases caused by bacterial infection. We used a mouse model of Salmonella Enteritidis infection, which included pretreatment with S. boulardii, to reveal the protection mechanisms of S. boulardii against Salmonella Enteritidis infection, including the translocation of Salmonella Enteritidis to the liver 10 days after Salmonella Enteritidis challenge, and the colonisation of Salmonella Enteritidis and the formation of hepatic tissue lesions in mice after Salmonella Enteritidis challenge on the 10th day. Compared with Salmonella Enteritidis infection in mice, S. boulardii decreased Salmonella Enteritidis translocation to the liver by 96%, and 99% of Salmonella Enteritidis colonised the cecum on the 10th day. Saccharomyces boulardii also abated hepatic tissue injury caused by the infiltration of neutrophilic granulocytes, lymphocytes, and plasmocytes by decreasing the translocation of Salmonella to the liver. These findings demonstrated that S. boulardii is an effective agent in the prevention of the hepatic injury induced by Salmonella Enteritidis infection in a mouse model.

[The structure of the epithelial surface of the gastrointestinal tract of pikas (Ochotona pallasi and O. dauurica, Lagomorpha, Ochotonidae): functional and species specificity].

The macro- and microrelief of the surface of the digestive tract mucosa of two pika species--Pallas's (Ochotona pallasi) and Daurian (O. dauurica)--were studied in detail using whole-mount preparations and scanning electron microscopy. The structural features of the intestinal mucosal surface specific of mammals, such as the formation of projections on the crest of the cecal spiral fold and microcells in the colonic ampulla, were studied. It was found that the colonic mucosa forms sparse large conical villi in pocket cavities and on the surface of muscle bands. Significant differences in the cecal mucosal relief were found between the species studied. The possible functional significance of the identified morphological features is discussed.

Cecorraphy in single layer using polypropylene and poliglecaprone 25 threads: comparative study in rats.

PURPOSE: To compare sutures with polypropylene and poliglecaprone 25 after partial cecotomy in rats. METHODS: Thirty six rats divided into two groups, A and B, of 18 animals; each group was also divided into three subgroups of six animals sacrificed at 4(th), 7(th) and 14(th) days after surgery. Were studied the mortality, morbidity, complications attributable to sutures, macroscopy, optical microscopy and measurement of hydroxyproline at the level of the suture. RESULTS: There were no deaths or wound complications such as hematoma, seroma, abscess, evisceration or eventration. On microscopic evaluation reepithelization, coaptation and inflammation in both groups did not differ significantly. The average rate of tissue hydroxyproline found in the samples on the 4(th) day after surgery, was 21.38 mg/g tissue for group A and 16.68 mg/g for group B; on day 7 after surgery, the average was 15.64 mg/g tissue for group A and 26.53 mg/g for group B; on day 14, the average was 8.09 mg/g tissue for group A and 25.07 mg/g for group B. CONCLUSION: There were no differences on clinical evolution, macroscopic aspect, microscopic data and hydroxyproline concentration on both sutures.

Cecorraphy in single layer using polypropylene and poliglecaprone 25 threads: comparative study in rats / Cecorrafia em plano único com fios de polipropileno e poliglecaprone 25: estudo comparativo em ratos

PURPOSE: To compare sutures with polypropylene and poliglecaprone 25 after partial cecotomy in rats. METHODS: Thirty six rats divided into two groups, A and B, of 18 animals; each group was also divided into three subgroups of six animals sacrificed at 4th, 7th and 14th days after surgery. Were studied the mortality, morbidity, complications attributable to sutures, macroscopy, optical microscopy and measurement of hydroxyproline at the level of the suture. RESULTS: There were no deaths or wound complications such as hematoma, seroma, abscess, evisceration or eventration. On microscopic evaluation reepithelization, coaptation and inflammation in both groups did not differ significantly. The average rate of tissue hydroxyproline found in the samples on the 4th day after surgery, was 21.38 mg/g tissue for group A and 16.68 mg/g for group B; on day 7 after surgery, the average was 15.64 mg/g tissue for group A and 26.53 mg/g for group B; on day 14, the average was 8.09 mg/g tissue for group A and 25.07 mg/g for group B. CONCLUSION: There were no differences on clinical evolution, macroscopic aspect, microscopic data and hydroxyproline concentration on both sutures.

OBJETIVO: Comparar a sutura com fio de polipropileno e poliglecaprone 25 após cecotomia parcial em ratos. MÉTODOS: Trinta e seis ratos foram distribuídos em dois grupos A e B de 18 animais, e cada grupo foi dividido em três subgrupos de seis, sacrificados no 4º, 7º e 14º dias do pós-operatório. Estudou-se a mortalidade, morbidade, complicações atribuíveis às suturas, macroscopia, microscopia ótica e dosagem de hidroxiprolina no nível da sutura. RESULTADOS: Não houve mortalidade ou complicações da ferida operatória como hematoma, seroma, abscesso, evisceração ou eventração. Na avaliação microscópica os critérios de re-epitelização, coaptação e processo inflamatório ambos os grupos não apresentaram diferença significativa. A taxa tecidual média da hidroxiprolina encontrada nas amostras no 4º dia de pós-operatório foi de 21,38 mg/g de tecido para o grupo A e 16,68 mg/g para o grupo B; no 7º dia a média foi de 15,64 mg/g de tecido para o grupo A e 26,53 mg/g para o grupo B; no 14º dia ela foi de 8,09 mg/g de tecido para o grupo A e 25,07 mg/g para o grupo B. CONCLUSÃO: Não houve diferença estatística entre a evolução clínica, avaliação macroscópica, microscopia e dosagem de hidroxiprolina entre as suturas realizadas com os fios estudados.

Three-dimensional demonstration of the interstitial cells of Cajal associated with the submucosal plexus in guinea-pig caecum.

The guinea-pig caecum was studied by using immunohistochemistry for Kit receptors and nerves to clarify whether interstitial cells of Cajal (ICC) were localized in association with the submucosal plexus (ICC-SP). A large area of the guinea-pig caecum was nearly devoid of longitudinal muscles, because they were concentrated into three bundles of the taenia caeci (coli) and this allowed clear observation of the myenteric and submucosal plexus as separate networks in whole-mount stretch preparations. The myenteric plexus was observed as a loose polygonal network consisting in elongated ganglia and long connecting nerve strands, whereas the submucosal plexus was identified as smaller ovoid ganglia connected to much thinner nerve strands in different tissue layers. Three-dimensional reconstruction of confocal images revealed multipolar-shaped ICC-SP located around the submucosal ganglion in a basket formation. Bipolar ICC-SP were also observed along the connecting nerve strands of the submucosal plexus. The functional involvement of ICC-SP in mucosal activity is discussed in relation to fluid transportation. This three-dimensional study of ICC-SP thus provides a candidate for the most suitable material available for functional experiments examining the physiological significance of ICC-SP.

Eimeria tenella: effects of diclazuril treatment on microneme genes expression in second-generation merozoites and pathological changes of caeca in parasitized chickens.

The effects of diclazuril on mRNA expression levels of invasion-related microneme genes were examined in second-generation merozoites of Eimeria tenella (E. tenella) by quantitative real-time (QRT) PCR. Diclazruil treatment of infected chickens significantly decreased the number of second-generation merozoites by 65.13%, and resulted in downregulation of EtMIC genes: EtMIC1 by 65.63%, EtMIC2 by 64.12%, EtMIC3 by 56.82%, EtMIC4 by 73.48%, and EtMIC5 by 78.17%. SEM images of caecum tissue from uninfected chickens showed regular intestinal villus structure. In infected chickens, a distinct loss of the superficial epithelium, with a flattened mucosa and large-area necrosis and anabrosis, was evident. In diclazruil-treated chickens, a decrease in merozoite number and a visibly improved appearance of the caeca were noted. These improvements appeared to be mediated in part by downregulation of the expression of invasion-related EtMIC genes in response to diclazuril.

The ultrastructural investigation of the midgut in the quill mite Syringophilopsis fringilla (Acari, Trombidiformes: Syringophilidae).

The midgut of the females of Syringophilopsis fringilla (Fritsch) composed of anterior midgut and excretory organ (=posterior midgut) was investigated by means of light and transmission electron microscopy. The anterior midgut includes the ventriculus and two pairs of midgut caeca. These organs are lined by a similar epithelium except for the region adjacent to the coxal glands. Four cell subtypes were distinguished in the epithelium of the anterior midgut. All of them evidently represent physiological states of a single cell type. The digestive cells are most abundant. These cells are rich in rough endoplasmic reticulum and participate both in secretion and intracellular digestion. They form macropinocytotic vesicles in the apical region and a lot of secondary lysosomes in the central cytoplasm. After accumulating various residual bodies and spherites, the digestive cells transform into the excretory cells. The latter can be either extruded into the gut lumen or bud off their apical region and enter a new digestive cycle. The secretory cells were not found in all specimens examined. They are characterized by the presence of dense membrane-bounded granules, 2-4 microm in diameter, as well as by an extensive rough endoplasmic reticulum and Golgi bodies. The ventricular wall adjacent to the coxal glands demonstrates features of transporting epithelia. The cells are characterized by irregularly branched apical processes and a high concentration of mitochondria. The main function of the excretory organ (posterior midgut) is the elimination of nitrogenous waste. Formation of guanine-containing granules in the cytoplasm of the epithelial cells was shown to be associated with Golgi activity. The excretory granules are released into the gut lumen by means of eccrine or apocrine secretion. Evacuation of the fecal masses occurs periodically. Mitotic figures have been observed occasionally in the epithelial cells of the anterior midgut.

Analysis of myenteric neurons of the cecum of diabetic rats after supplementation with ascorbic acid / Análisis de neuronas mientéricas del ciego de ratas diabéticas después de suplementación con ácido ascórbico

The objective of this work was to investigate the neuroprotective action of the ascorbic acid over the myenteric neurons in the cecum of Wistar rats, four months after induction of the diabetes mellitus experimental with streptozotocin. Three groups with five rats each were used: C- controls, D- diabetic, DA- diabetic treated with ascorbic acid. For evidentiation of the myenteric neurons was carried out to Giemsa's technique. Were evaluated the areas of cell bodies of 500 neurons in each group studied. The quantitative analysis was carried out in an area of 16.6 mm2 in each cecum studied. In the animals diabetic observed elevation of the glycemia and glycated hemoglobin. The supplementation with ascorbic acid was effective under the myenteric neurons of the cecum of diabetics rays, since was presented the effect neuroprotective and neurotrofic.

El objetivo de este trabajo fue verificar el efecto neuroprotector del ácido ascórbico sobre las neuronas mientéricas en el ciego de Rattus Wistar, cuatro meses después de la inducción de diabetes mellitus experimental con estreptozotocina. Utilizamos tres grupos de animales: C- control, D- diabético, DA- diabético tratado con ácido ascórbico. Para la observación de las neuronas mientéricas fue llevado a cabo la técnica de Giemsa. Fueron evaluadas las áreas del soma de 500 neuronas, en cada grupo estudiado. El análisis cuantitativo fue llevado a cabo, en cada ciego, en un área de 16,6 mm². En los animales diabéticos, se observó la elevación de la glicemia y de la hemoglobina glicosilada. La suplementación con ácido ascórbico fue efectiva en las neuronas mientéricas del ciego de animales diabéticos, ya que se produjeron los efectos neuroprotetor y neurotrófico.