scholarly journals Avian intestinal ultrastructure changes provide insight into the pathogenesis of enteric diseases and probiotic mode of action

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Shaniko Shini ◽  
R. Claire Aland ◽  
Wayne L. Bryden
Keyword(s):  
Electron Microscopy ◽  
Villous Atrophy ◽  
Adherens Junction ◽  
Necrotic Enteritis ◽  
Intestinal Immunity ◽  
Gut Health ◽  
Ileal Mucosa ◽  
Enteric Diseases ◽  
Transmission Electron ◽  
Intestinal Infections

AbstractEpithelial damage and loss of barrier integrity occur following intestinal infections in humans and animals. Gut health was evaluated by electron microscopy in an avian model that exposed birds to subclinical necrotic enteritis (NE) and fed them a diet supplemented with the probiotic Bacillus amyloliquefaciens strain H57 (H57). Scanning electron microscopy of ileal mucosa revealed significant villus damage, including focal erosions of epithelial cells and villous atrophy, while transmission electron microscopy demonstrated severe enterocyte damage and loss of cellular integrity in NE-exposed birds. In particular, mitochondria were morphologically altered, appearing irregular in shape or swollen, and containing electron-lucent regions of matrix and damaged cristae. Apical junctional complexes between adjacent enterocytes were significantly shorter, and the adherens junction was saccular, suggesting loss of epithelial integrity in NE birds. Segmented filamentous bacteria attached to villi, which play an important role in intestinal immunity, were more numerous in birds exposed to NE. The results suggest that mitochondrial damage may be an important initiator of NE pathogenesis, while H57 maintains epithelium and improves the integrity of intestinal mucosa. Potential actions of H57 are discussed that further define the mechanisms responsible for probiotic bacteria’s role in maintaining gut health.

scholarly journals Intestinal Lesions in Specific-Pathogen-Free Lambs Associated with a Cryptosporidium from Calves with Diarrhea

1982 ◽  
Vol 19 (1) ◽  
pp. 67-78 ◽  
Author(s):  
K. W. Angus ◽  
S. Tzipori ◽  
E. W. Gray
Keyword(s):  
Electron Microscopy ◽  
Lamina Propria ◽  
Villous Atrophy ◽  
Post Inoculation ◽  
Specific Pathogen Free ◽  
Intestinal Lesions ◽  
Transmission Electron ◽  
Specific Pathogen ◽  
Small Intestinal ◽  
Neutrophil Leukocytes

Small and large intestines of seven specific pathogen-free lambs infected with cryptosporidia from calves with diarrhea were examined by scanning and transmission electron microscopy and by light microscopy. The small intestine was infected in all the lambs, and the cecum and colon in three. Small intestinal alterations were severe villous atrophy and dilatation of the crypts of Lieberkühn. Epithelial cross-bridging between contiguous villi caused much villous fusion. Epithelial cells constituting the bridges were connected by desmosomal junctions, and were continuous with the epithelial coverings of the associated villi. The lamina propria was heavily infiltrated with neutrophil leukocytes. Infected crypts in cecum and colon were dilated and devoid of mucus-secreting cells, while the ridges between crypts were hypertrophied, and the lamina propria was infiltrated by neutrophils. Cell vegetations with adherent bacteria were present in the surface intestinal epithelium of two lambs infected for 11 and 14 days, respectively. No adherent bacteria were seen in any site in lambs killed up to six days post-inoculation.

scholarly journals Outbreak of cryptosporidiosis in African penguins Spheniscus demersus

2020 ◽  
Vol 140 ◽  
pp. 143-149
Author(s):  
R Hurtado ◽  
NJ Parsons ◽  
TA Gous ◽  
Sv der Spuy ◽  
R Klusener ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Clinical Signs ◽  
Necrotic Enteritis ◽  
Rehabilitation Center ◽  
Transmission Electron ◽  
Petechial Hemorrhage ◽  
Spheniscus Demersus ◽  
Undigested Food ◽  
African Penguin

Cryptosporidium spp. are parasitic intracellular protozoa that infect the digestive, respiratory, and urinary tracts of vertebrates. The disease affects many different avian species across all continents, and >25 species and genotypes of Cryptosporidium have been documented infecting birds. We report on an outbreak of cryptosporidiosis in African penguin Spheniscus demersus chicks admitted to a rehabilitation center in South Africa from February 2012 to October 2013. Eighteen cases were confirmed through histopathology. The most frequent clinical signs were regurgitation (78%), dyspnea (72%), decreased weight gain or weight loss (72%), and lethargy (50%). Clinical signs began 8-46 d after hatching or admission (median: 13 d), and death followed 1-41 d after the onset of clinical signs (median: 13.5 d). The most frequent necropsy findings were stomach distended with undigested food or gas (78%), mildly reddened lungs (56%), spleen petechial hemorrhage (44%), and kidney congestion (39%). The most frequent histopathological findings were necrotic bursitis (89%), necrotic enteritis (83%), and bursal atrophy (67%). Small round or oval basophilic bodies (3-5 µm diameter) consistent with Cryptosporidium sp. were closely associated with the surface of the epithelial cells or in the lumen of the bursa (89%), large intestine (61%), small intestine (44%), trachea (22%), and ventriculus (6%). Transmission electron microscopy of 1 case confirmed that these organisms were Cryptosporidium sp. To our knowledge, this is the first report of cryptosporidiosis in penguins, raising concern of the potential implications for the conservation of these species.

scholarly journals Clostridium perfringens produces an adhesive pilus required for the pathogenesis of necrotic enteritis in poultry

2021 ◽  
Author(s):  
D. Lepp ◽  
Y. Zhou ◽  
S. Ojha ◽  
I. Mehdizadeh Gohari ◽  
J. Carere ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Cell Surface ◽  
Clostridium Perfringens ◽  
Direct Evidence ◽  
Necrotic Enteritis ◽  
Null Mutant ◽  
Transmission Electron ◽  
Mutant Strains ◽  
Important Disease

Clostridium perfringens Type G strains cause necrotic enteritis (NE) in poultry, an economically important disease that is a major target of in-feed antibiotics. NE is a multifactorial disease, involving not only the critically-important NetB toxin, but also additional virulence and virulence-associated factors. We previously identified a C. perfringens chromosomal locus (VR-10B) associated with disease-causing strains that is predicted to encode a sortase-dependant pilus. In the current study, we sought to provide direct evidence for the production of a pilus by C. perfringens and establish its role in NE pathogenesis. Pilus structures in virulent C. perfringens strain CP1 were visualized by transmission electron microscopy (TEM) of immuno-gold labelled cells. Filamentous structures were observed extending from the cell surface in wild-type CP1, but not from isogenic pilin-null mutant strains. In addition, immuno-blotting of cell surface proteins demonstrated that CP1, but not the null mutant strains, produced a high molecular weight ladder-like pattern characteristic of a pilus polymer. Binding to collagen types I, II and IV was significantly reduced (Tukey’s; p<0.01) in all three pilin mutants compared to CP1, and could be specifically blocked by CnaA and FimA antisera, indicating that these pilins participate in adherence. Furthermore, both fimA and fimB null mutants were both severely attenuated in their ability to cause disease in an in vivo chicken NE challenge model. Together, these results provide the first direct evidence for the production of a sortase-dependant pilus by C. perfringens, and confirm its critical role in NE pathogenesis and collagen-binding. Importance In necrotic enteritis (NE), an intestinal disease of chickens, Clostridium perfringens cells adhere tightly to damaged intestinal tissue, but the factors involved are not known. We previously discovered a cluster of C. perfringens genes predicted to encode a pilus, a hair-like bacterial surface structure commonly involved in adherence. In the current study, we have directly imaged this pilus using transmission electron microscopy (TEM). We also show that inactivation of the pilus genes stops pilus production, significantly reducing the bacterium's ability to bind collagen and cause disease. Importantly, this is the first direct evidence for the production of a sortase-dependant pilus by C. perfringens, revealing a promising new target for developing therapeutics to combat this economically important disease.

scholarly journals Spontaneous Cryptosporidiosis in an Adult Female Rabbit

1979 ◽  
Vol 16 (1) ◽  
pp. 89-95 ◽  
Author(s):  
L. R. Inman ◽  
A. Takeuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Adult Female ◽  
Ileal Mucosa ◽  
Double Membrane ◽  
Female Rabbit ◽  
Transmission Electron ◽  
Asymptomatic Adult ◽  
Scanning Electron

An asymptomatic adult female rabbit had intestinal cryptosporidiosis. The ileum had blunted villi, a decrease in villus-crypt ratio and a mild edema in the lamina propria. Transmission electron microscopy showed the parasite to be a Cryptosporidium similar to those reported in mouse, guinea pig, lamb, calf, horse, monkey and man. This organism is referred to as Cryptosporidium cuniculus. Scanning electron microscopy on ileal mucosa showed altered intestinal microvilli in the attachment of the cryptosporidia. It is postulated that the organism was enveloped by the microvilli of the ileal epithelial cells which then fused and formed a continuous double membrane around the parasite.

Techniques for Transmission Electron Microscopy of Fiber-Matrix Interfaces in Aluminum Alloy-Boron Composites by Ion Erosio

1971 ◽  
Vol 29 ◽  
pp. 204-205
Author(s):  
G. G. Shaw
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Aluminum Alloy ◽  
Electron Beam ◽  
Pure Aluminum ◽  
Ion Milling ◽  
Transmission Electron ◽  
Fiber Matrix ◽  
Ion Erosion ◽  
Row Of Fibers

The morphology and composition of the fiber-matrix interface can best be studied by transmission electron microscopy and electron diffraction. For some composites satisfactory samples can be prepared by electropolishing. For others such as aluminum alloy-boron composites ion erosion is necessary.When one wishes to examine a specimen with the electron beam perpendicular to the fiber, preparation is as follows: A 1/8 in. disk is cut from the sample with a cylindrical tool by spark machining. Thin slices, 5 mils thick, containing one row of fibers, are then, spark-machined from the disk. After spark machining, the slice is carefully polished with diamond paste until the row of fibers is exposed on each side, as shown in Figure 1.In the case where examination is desired with the electron beam parallel to the fiber, preparation is as follows: Experimental composites are usually 50 mils or less in thickness so an auxiliary holder is necessary during ion milling and for easy transfer to the electron microscope. This holder is pure aluminum sheet, 3 mils thick.

Direct Observation of Heat Exchanger Deposits by Cross Sectioning

1967 ◽  
Vol 25 ◽  
pp. 364-365
Author(s):  
R. W. Anderson ◽  
D. L. Senecal
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Heat Exchanger ◽  
Direct Observation ◽  
Cross Sections ◽  
Preparation Method ◽  
Specimen Preparation ◽  
Flowing Water ◽  
Transmission Electron ◽  
Deposit Layer

A problem was presented to observe the packing densities of deposits of sub-micron corrosion product particles. The deposits were 5-100 mils thick and had formed on the inside surfaces of 3/8 inch diameter Zircaloy-2 heat exchanger tubes. The particles were iron oxides deposited from flowing water and consequently were only weakly bonded. Particular care was required during handling to preserve the original formations of the deposits. The specimen preparation method described below allowed direct observation of cross sections of the deposit layers by transmission electron microscopy.The specimens were short sections of the tubes (about 3 inches long) that were carefully cut from the systems. The insides of the tube sections were first coated with a thin layer of a fluid epoxy resin by dipping. This coating served to impregnate the deposit layer as well as to protect the layer if subsequent handling were required.

Phase Transformations in Ti-7w/oMo-3w/oMe Alloy

1974 ◽  
Vol 32 ◽  
pp. 514-515
Author(s):  
S. Fujishiro
Keyword(s):  
Electron Microscopy ◽  
Mechanical Properties ◽  
Transmission Electron Microscopy ◽  
Tensile Strength ◽  
Mechanical Processing ◽  
Ray Method ◽  
X Ray ◽  
Transmission Electron ◽  
Water Quench ◽  
Alpha Beta

The mechanical properties of three titanium alloys (Ti-7Mo-3Al, Ti-7Mo- 3Cu and Ti-7Mo-3Ta) were evaluated as function of: 1) Solutionizing in the beta field and aging, 2) Thermal Mechanical Processing in the beta field and aging, 3) Solutionizing in the alpha + beta field and aging. The samples were isothermally aged in the temperature range 300° to 700*C for 4 to 24 hours, followed by a water quench. Transmission electron microscopy and X-ray method were used to identify the phase formed. All three alloys solutionized at 1050°C (beta field) transformed to martensitic alpha (alpha prime) upon being water quenched. Despite this heavily strained alpha prime, which is characterized by microtwins the tensile strength of the as-quenched alloys is relatively low and the elongation is as high as 30%.

Ultrastructural Analysis of the Salivary Glands of Gromphadorhina Portentosa (Schaum)

1977 ◽  
Vol 35 ◽  
pp. 638-639
Author(s):  
P.J. Dailey
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Salivary Glands ◽  
Secretory Vesicles ◽  
The Past ◽  
Transmission Electron ◽  
Means Of Transmission ◽  
Gromphadorhina Portentosa ◽  
Scanning Electron ◽  
Topographical Relief

The structure of insect salivary glands has been extensively investigated during the past decade; however, none have attempted scanning electron microscopy (SEM) in ultrastructural examinations of these secretory organs. This study correlates fine structure by means of SEM cryofractography with that of thin-sectioned epoxy embedded material observed by means of transmission electron microscopy (TEM).Salivary glands of Gromphadorhina portentosa were excised and immediately submerged in cold (4°C) paraformaldehyde-glutaraldehyde fixative1 for 2 hr, washed and post-fixed in 1 per cent 0s04 in phosphosphate buffer (4°C for 2 hr). After ethanolic dehydration half of the samples were embedded in Epon 812 for TEM and half cryofractured and subsequently critical point dried for SEM. Dried specimens were mounted on aluminum stubs and coated with approximately 150 Å of gold in a cold sputtering apparatus.Figure 1 shows a cryofractured plane through a salivary acinus revealing topographical relief of secretory vesicles.

Two- or three-way observations of the identical cell elements within the same sections by LM, TEM and/or SEM

1978 ◽  
Vol 36 (2) ◽  
pp. 82-83 ◽  
Author(s):  
Nakazo Watari ◽  
Yasuaki Hotta ◽  
Yoshio Mabuchi
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Light Microscopy ◽  
Thin Sections ◽  
Transmission Electron ◽  
Identical Cell ◽  
Electron Microscopes ◽  
Scanning Electron

It is very useful if we can observe the identical cell elements within the same sections by light microscopy (LM), transmission electron microscopy (TEM) and/or scanning electron microscopy (SEM) sequentially, because, the cell fine structure can not be indicated by LM, while the color is; on the other hand, the cell fine structure can be very easily observed by EM, although its color properties may not. However, there is one problem in that LM requires thick sections of over 1 μm, while EM needs very thin sections of under 100 nm. Recently, we have developed a new method to observe the same cell elements within the same plastic sections using both light and transmission (conventional or high-voltage) electron microscopes.In this paper, we have developed two new observation methods for the identical cell elements within the same sections, both plastic-embedded and paraffin-embedded, using light microscopy, transmission electron microscopy and/or scanning electron microscopy (Fig. 1).

Transmission Electron Microscopy Studies of Pore Cells in Limax Maximus

1977 ◽  
Vol 35 ◽  
pp. 656-657
Author(s):  
B. S. Beltz
Keyword(s):  
Electron Microscopy ◽  
Cell Periphery ◽  
Salivary Duct ◽  
Terrestrial Mollusc ◽  
Buccal Ganglia ◽  
Transmission Electron ◽  
Pore Cells ◽  
Gland Tissue ◽  
Storage Area ◽  
Limax Maximus

The cells which are described in this study surround the salivary nerve of the terrestrial mollusc, Limax maximus. The salivary system of Limax consists of bilateral glands, ducts, and nerves. The salivary nerves originate at the buccal ganglia, which are situated on the posterior face of the buccal mass, and run along the salivary duct to the gland. The salivary nerve branches several times near the gland, and eventually sends processes into the gland.The pore cells begin to appear at the first large branch point of the salivary nerve, near the gland (Figure 1). They follow the nerve distally and eventually accompany the nerve branches into the gland tissue. The cells are 20-50 microns in diameter and contain very small nuclei (1-5 microns) (Figure 2).The cytoplasm of the pore cells is segregated into a storage area of glycogen and an organelle region located in a band around the cell periphery (Figure 3).

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