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1.
Pregnant goats were given Brucella abortus intravenously or in uterine arteries, and tissues from the uterus and placentae were examined at various post-inoculation intervals to study mechanisms of placental infection. Placentitis was present by 5 days post-inoculation and abortions occurred within 11 days. B. abortus was identified in placentae by light microscopy and immunoperoxidase techniques. B. abortus was first seen in erythrophagocytic trophoblasts of the placentome. Subsequently, high numbers of B. abortus were seen in periplacentomal chorioallantoic trophoblasts. Trophoblast necrosis, chorioallantoic ulceration, and large numbers of B. abortus in chorionic villi were present in later stages of infection. These results suggest that entry and replication of B. abortus in trophoblasts precede placentome and fetal infection and that trophoblasts are the source of B. abortus for these tissues. Experimental caprine brucellosis closely resembles bovine and ovine brucellosis and it provides a model to study the intracellular development of B. abortus in trophoblasts.  相似文献   

2.
Uptake, transfer to rough endoplasmic reticulum, and intracellular growth of Brucella abortus were studied in Vero cells treated with endocytic and metabolic inhibitors. Infection of Vero cells was suppressed when inhibitors of energy metabolism (iodoacetate, dinitrophenol), receptor-mediated endocytosis (monodansylcadaverine, amantadine, methylamine), or endosomal acidification (chloroquine, ammonium chloride, monensin) were added to the inoculum. Inhibition was not observed when these drugs were added after the inoculation period. Infection of Vero cells by B abortus was inhibited by dibutyryl-cyclic adenosine monophosphate and Vibrio cholerae enterotoxin, but was stimulated by dibutyryl-cyclic guanosine monophosphate and escherichia coli heat-stable enterotoxin a. Uptake of B abortus by Vero cells was not prevented by colchicine, but was abolished by cytochalasin B. Uptake of heat-killed B abortus and noninvasive E coli was similar to that of viable brucellae. Intracellular growth of B abortus was not affected by cycloheximide. Results indicate that: B abortus may be internalized by a receptor-mediated phagocytic process; transfer of B abortus from phagosomes to rough endoplasmic reticulum may require endosomal acidification; and replication of B abortus within the rough endoplasmic reticulum may not depend on protein synthesis by the host cell.  相似文献   

3.
Pathogenesis of Brucella abortus in chicken embryos   总被引：2，自引：0，他引：2  
Chicken embryos inoculated with Brucella abortus at 6, 10, and 12 days of incubation were examined by light and electron microscopy. B. abortus was identified by avidin-biotin immunoperoxidase and immunogold techniques. Death occurred from 2 to 5 days post-inoculation, depending on age of the embryo and route of inoculation. B. abortus was recovered from all infected eggs. Brucellae had spread throughout all tissues and localized preferentially within cells of mesodermal derivation. Organ distribution and degree of bacterial replication varied with age of the embryo at time of inoculation. In 6-day-old embryos, B. abortus localized preferentially in endoderm and mesoderm of yolk sac wall, extra- and intraembryonic serosal epithelia, and glomeruli of the mesonephros. In 10- and 12-day-old embryos, B. abortus spread to all tissues; renal glomeruli, liver, spleen, and heart were most severely infected. Intracellular B. abortus was within the rough endoplasmic reticulum of mesenchymal, mesothelial, yolk endodermal, and hepatic cells. In mononuclear phagocytes, endothelial cells, and granulocytes, bacteria were within membrane-bound vacuoles. Intracellular replication of B. abortus in embryonic tissues, especially yolk endoderm, closely resembled that in experimental infections of trophoblasts.  相似文献   

4.
Vero cells were inoculated with the six species of Brucella (B. abortus, B. melitensis, B. suis, B. neotomae, B. canis, and B. ovis) and examined by fluorescence and electron microscopy. All Brucella spp. were internalized by Vero cells. In all cells except those inoculated with B. canis, the numbers of intracellular brucellae increased with time after inoculation. Intracellular brucellae were first seen within phagosomes and phagolysosomes. Subsequent localization within cisternae of the rough endoplasmic reticulum was seen with all species of Brucella, except B. canis, which was restricted to phagolysosomes. Although rough brucellae were more adherent and entered a greater number of Vero cells, intracellular replication occurred in a larger percentage of cells with smooth rather than with rough brucellae. These results suggest that phagocytosed Brucella spp. are transferred 1) to cisternae of the rough endoplasmic reticulum, where unrestricted bacterial replication takes place; or 2) to phagolysosomes in which Brucella spp. fail to replicate. The various strains of Brucella spp. differ in their ability to induce their own transfer to the rough endoplasmic reticulum.  相似文献   

5.
We describe here the gross and microscopic lesions in 18 experimentally induced and 120 natural Campylobacter abortions. In natural Campylobacter abortions, gross lesions were reported infrequently; placentitis was recorded in 6% and hepatic lesions in 4% of our field cases. Placentitis was the microscopic lesion identified most consistently in natural abortions (93%) and was often observed in association with abundant bacterial colonies in chorionic villi (54%) and less often with placental vasculitis (13%). In natural abortions, suppurative fetal pneumonia (48%), necrosuppurative hepatitis (16%), and purulent meningitis (7%) were also observed. The better-preserved specimens from experimentally induced abortions were utilized to define placental changes more precisely. Placentitis was identified in all 18 experimentally induced abortions and was observed most consistently in the chorionic villus stroma (100%), often accompanied by suppurative surface exudate (89%). An inflammatory infiltrate was less commonly identified in the cotyledonary hilus (39%) and intercotyledonary placenta (22%). Bacteria were visualized in H&E-stained sections in 89% of placentas from experimentally infected ewes, primarily as well-demarcated bacterial colonies within subtrophoblastic, sinusoidal capillaries (89%), in the cotyledonary villus stroma (89%), and within the cytoplasm of trophoblasts (22%). Transmission electron microscopy and immunohistochemistry confirmed that the vast majority of the well-demarcated bacterial colonies characteristic of Campylobacter abortion were within subtrophoblastic sinusoidal capillaries. The most characteristic microscopic lesions identified in cases of Campylobacter abortion in sheep were placentitis with placental bacterial colonies, placental vasculitis, and fetal pneumonia.  相似文献   

6.
OBJECTIVE: To determine the virulence of a Brucella abortus mutant, BA25, lacking a major 25 kd outer membrane protein (Omp25) in cattle. ANIMALS: 20 mixed-breed heifers in late gestation. PROCEDURE: 10 heifers were inoculated with 1 x 10(7) colony-forming units of the Omp25 mutant via the conjunctival sac, and an equal number were infected with the virulent parental strain B. abortus 2308. The delivery status of the dams was recorded, and colonization was assessed following necropsy. The ability of BA25 to replicate inside bovine phagocytes and chorionic trophoblasts was also evaluated in vitro because of the propensity of virulent brucellae to replicate inside these cells in vivo. RESULTS: The parental strain induced abortions in 5 of 10 inoculated cattle, whereas only 1 of 10 dams exposed to BA25 aborted. Brucella abortus strain 2308 colonized all of the cow-calf pairs and induced Brucella-specific antibodies in 100% of the dams. In contrast, BA25 was isolated by bacteriologic cultural technique from 30% of the calves and 50% of the inoculated dams (n = 10). Of the 10 heifers inoculated with BA25, 4 did not develop Brucella-specific antibodies nor were they colonized by the mutant strain. In bovine macrophages and chorionic trophoblasts, BA25 replicated in significantly lower numbers than the virulent parental strain (n = 3). CONCLUSIONS AND CLINICAL RELEVANCE: The 25 kd outer membrane protein may be an important virulence factor for B. abortus in cattle. The attenuation of the Omp25 mutant in cattle may involve the inability of BA25 to replicate efficiently in bovine phagocytes and chorionic trophoblasts.  相似文献   

7.
It was shown in this study that complement-resistant Brucella abortus used were unable to activate complement in the absence of specific antibody. Complement-resistant isolates possessed O-antigen, but complement-sensitive organisms used are O-antigen deficient. Since B. abortus LPS does not activate the alternative pathway of complement, we concluded that activation of bovine complement must be due to some other mechanism. In this study, it was shown that bovine C1 binds to the outer membrane proteins of B. abortus. Isolated outer membrane proteins of both smooth (O-antigen positive) and rough (O-antigen negative) B. abortus used bind to C1q. However, only rough isolates were killed by complement. All of the O-antigen positive B. abortus isolates were complement-resistant. We propose that O-antigen shields outer membrane proteins and blocks C1q binding.  相似文献   

8.
Brucella abortus infection has not been documented in llamas. This report describes the abortion of the only pregnant animal in a group of 12. The llama was infected by inoculating 1 x 10(8) viable B. abortus organisms into the conjunctival sac. Forty-three days postinfection, the llama aborted a fetus of approximately 8 months gestational age. Brucella organisms were isolated from the placenta and all fetal specimens examined. These organisms were also isolated from the dam's mammary gland and numerous lymph nodes when the llama was necropsied 42 days later. Microscopically, there was a moderate, multifocal, lymphocytic and histiocytic, subacute placentitis with marked loss of trophoblastic epithelial cells. The superficial chorioallantoic stroma contained abundant necrotic and mineralized debris as well as numerous swollen capillaries protruding multifocally from the denuded surface. Immunohistochemistry revealed that these capillaries, as well as sloughed and intact trophoblasts, were expanded by large numbers of Brucella organisms. Brucellar antigen was also detected in occasional macrophages in the fetal kidney and lung. Ultrastructurally, bacteria labeled by an antibody-based colloidal gold procedure were located within degenerate capillaries, within necrotic leukocytes, and extracellularly in the placental stroma.  相似文献   

9.
The structure and functions of placentas were examined in 3 species of rorqual whales, common minke (Balaenoptera acutorostrata), Bryde’s (B. brydei) and sei (B. borealis) whales, with the aim of confirming the structural characteristics of the chorion, including the presence of the areolar part, and clarifying steroidogenic activities and fetomaternal interactions in the placentas of these whales. Placentas were collected from the second phase of the Japanese Whale Research Program under Special Permit in the North Pacific (JARPN II). Histological and ultrastructural examinations revealed that these whale placentas were epitheliochorial placentas with the interdigitation of chorionic villi lined by monolayer uninucleate cells (trophoblast cells) and endometrial crypts as well as folded placentation by fold-like chorionic villi. Moreover, well-developed pouch-like areolae were observed in the placentas, and active absorption was suggested in the chorionic epithelial cells of the areolar part (areolar trophoblast cells). Berlin blue staining showed the presence of ferric ions (Fe3+) in the uterine glandular epithelial cells and within the stroma of chorionic villi in the areolar part. An immunohistochemical examination revealed tartrate-resistant acid phosphatase (TRAP; known as uteroferrin in uteri) in the cytoplasm of glandular cells and areolar trophoblast cells. This result suggested that, in cetaceans, uteroferrin is used to supply iron to the fetus. Furthermore, immunoreactivity for P450scc and P450arom was detected in trophoblast cells, but not in areolar trophoblast cells, suggesting that trophoblast cells synthesize estrogen in whale placentas. Therefore, we herein immunohistochemically revealed the localization of aromatase and uteroferrin in cetacean placentas during pregnancy for the first time.  相似文献   

10.
In this study, we examined the existence and structure of areolae and the steroidogenesis of areolar trophoblast cells in the Antarctic minke whale placenta morphologically and immunohistochemically. Placentas were collected from the 15th, 16th and 18th Japanese Whale Research Program under Special Permit in the Antarctic (JARPA) and 1st JARPA II organized by the Institute of Cetacean Research in Tokyo, Japan. The opening and cavity of fetal areolae formed by taller columnar trophoblast cells (areolar trophoblast cells) with long microvilli and a bright cytoplasm, as compared with the trophoblast cells of the chorionic villi interdigitating with the endometrial crypts, were recognized in observations of serial sections. The opening of the areolar cavity was hidden by chorionic villi with areolar trophoblast cells. Furthermore, a closed pouch-like structure lined by tall columnar cells similar to areolar trophoblast cells within the stroma of chorionic villi was noticed and continued to the areolar cavity, with the opening seen on serial sections. In a surface investigation of the chorion and endometrium by SEM, maternal (endometrial) areolae irregularly surrounded by endometrial folds were obvious. Moreover, we distinguished areolar trophoblast cells with long microvilli attached with many blebs from trophoblast cells. In our immunohistochemical observations, a steroidogenic enzyme, cytochrome P450 side chain cleavage enzyme (P450scc), was detected with strong immunoreactivity in trophoblast cells. However, areolar trophoblast cells showed weak or no immunoreactivity for P450scc.  相似文献   

11.
Brucella is responsible for one of the major worldwide zoonoses. Over the last century, several vaccines have been used against brucellosis. Among these, the rough vaccine Brucella abortus RB51 was introduced with the idea that it would not interfere with the diagnosis of brucellosis. Recently, RB51 has been isolated from milk and vaginal exudates from vaccinated cows, thus raising the possibility of extensive bacterial replication in these animals. We hypothesized that shedding of RB51 might be related to a change in its intracellular cell cycle. Therefore, we have compared the intracellular trafficking in CHO cells of the virulent B. abortus 2308 and two RB51 strains, the vaccinal strain and the one isolated from cow milk. Both RB51 strains were transiently observed in phagosomes characterized by the presence of the early endosomal marker EEA1 and then were found in cathepsin D-enriched lysosomal compartments, in which they eventually underwent degradation at later post-infection times. In contrast, the virulent 2308 strain replicated within the endoplasmic reticulum. These results suggest that a change in intracellular trafficking cannot account for Brucella shedding in adult vaccinated cows.  相似文献   

12.
The present study represents the first comprehensive investigation of the glandular chambers of the placenta of the bitch. We examined the glandular chambers by SEM, TEM, ultrahistochemical and lectinhistochemical methods. The glandular epithelium is a high columnar epithelium with club-shaped apical protrusions bearing short microvilli. The epithelium forms extensive folds which fill up most of the lumen of the glandular chambers. Proceeding to the placental labyrinth, the glandular chambers are covered by the tips of the chorionic tufts. The trophoblast and the glandular epithelium are separated by a thick layer of secretions. The ultrastructure cytology of the columnar epithelium is characterized by several Golgi complexes and abundant apical-located mucus vesicles with a positive dialysed iron reaction. Lectin histochemistry reveals in general a strong reaction of these mucus vesicles with all lectins used. The mucus in the lumen of the glandular chamber reacts strongly with WGA. NeuWGA, LPA and ConA. The trophoblastic villi projecting into the lumen of the glandular chambers are covered by a pseudostratified epithelium consisting of a flat basal layer and a superficial columnar one. Its cytoplasm is filled with large fusing vacuoles containing longish screw-shaped structures. No morphological equivalent of absorption is found. According to the ultrastructure of the trophoblast, the significance of the mucus in the glandular chambers in stopping the invasive growth of the trophoblast is discussed.  相似文献   

13.
Chlamydophila abortus infects the placental trophoblast in sheep, humans and mice, causing cell damage and inflammation that culminates in abortion. Host control of C. abortus appears to be heavily dependant on interferon (IFN)-gamma production. IFN-gamma induces expression of the enzyme indoleamine 2,3-dioxygenase (IDO), resulting in the degradation of intracellular pools of tryptophan, thereby depriving the organism of this essential growth nutrient. The anti-chlamydial effects of IFN-gamma can be reversed by the addition of exogenous tryptophan. This finding is consistent with studies of the C. abortus genome sequence that have revealed that the organism lacks the capability to synthesise tryptophan from host cell substrates and is therefore dependant on host tryptophan. This raises an interesting paradox since the placental trophoblast in humans and mice is known to constitutively express IDO and degrade tryptophan, a phenomenon that has been linked to maternal immunological tolerance of the semi-allogeneic fetus. This paradox is discussed in the context of immune modulation during pregnancy, tryptophan biosynthesis by Chlamydiaceae and differences in placental structures between sheep, humans and mice.  相似文献   

14.
Placentitis, premature birth, and perinatal death were associated with Encephalitozoon cuniculi infection in an alpaca. Histologically, chorionic trophoblasts contained many Gram-positive, period acid-Schiff positive, variably acid-fast spores. Multifocal necrosis and infiltration by lymphocytes, eosinophils, and neutrophils were scattered throughout the chorionic membrane. Spores in trophoblasts were approximately 1 microm x 2 microm, thick-walled, and contained polar filaments and polar vacuoles consistent with microsporidia. The presence of E. cuniculi DNA was confirmed by sequencing the polymerase chain reaction amplicon from frozen placental tissue. A few glial nodules were scattered throughout the cerebrum, and mild lymphocytic inflammation was present in the heart, liver, and lung. No organisms were detected in tissues other than the placenta. This is the first reported case of E. cuniculi infection in an alpaca.  相似文献   

15.
The fine structure of plasma cells from the hemal lymph nodes of five healthy sheep was examined. The plasma cells showed many cytoplasmic processes, indicating motility and phagocytic activity. They were randomly located in the medullary cords of the hemal lymph nodes. Their perinuclear spaces were irregular and contained granular material similar in density to that of rough endoplasmic reticulum. Some of the plasma cells provided morphological evidence of erythrophagocytosis at different levels. Such a phenomenon was previously reported only in pathological conditions. The red blood cell being engulfed by the plasma cell appeared smaller than normal. Most of the examined plasma cells showed Russell's Bodies inside the cisternae of the rough endoplasmic reticulum of the healthy hemal lymph nodes. Some of these Russell's Bodies had fuzzy borders. They were separated from the rough endoplasmic reticulum by evenly spaced halos.
Many plasma cells showed physical contacts with mast cells in the form of cell junctions, and some showed cytoplasmic continuity between the two cells. Such relations have been discussed in this article.  相似文献   

16.
Ko KY  Kim JW  Her M  Kang SI  Jung SC  Cho DH  Kim JY 《Veterinary microbiology》2012,156(3-4):374-380
To overcome the limitations of serological diagnosis, including false positive reactions caused by other pathogens, specific antigens for diagnosis of brucellosis other than LPS have been required. The present study was conducted to separate and identify immuno-dominant insoluble proteins of Brucella abortus against the antisera of cattle infected with B. abortus, or/and Yersinia enterocolitica, or the sera of non-infected cattle. After separating insoluble proteins of B. abortus by two dimensional electrophoresis (2-DE), their immuno-reactivity was determined by western blotting. A portion of the immunogenic spots against the positive antisera of B. abortus that have the potential for use as specific antigens were identified by MS/MS analysis. Overall, 18 immunogenic insoluble proteins of B. abortus 1119-3 showed immuno-reactivity against only the positive antisera of B. abortus, but failed to have immunogenicity toward both the positive sera of Y. enterocolitica and the negative sera of B. abortus. Identification of these proteins revealed the following: F0F1 ATP synthase subunit β, solute-binding family 5 protein, 28 kDa OMP, Leu/Ile/Val-binding family protein, Histidinol dehyddrogenase, Hypothetical protein, Twin-arginine translocation pathway signal sequence domain-containing protein, Dihydroorotase, Serine protease family protein, β-hydroxyacyl-(acyl-carrier-protein) dehydratase FabA, Short-chain dehydrogenase-/reductase carbonic anhydrase, Orinithine carbamoyltransferase, Leucyl aminopeptidase, Cold shock DNA-binding domain-containing protein, Cu/Zn superoxide dismutase, and Methionine aminopeptidase. The 18 immunogenic proteins separated in the present study can be considered candidate antigens to minimize cross reaction in the diagnosis of brucellosis and useful sources for Brucella vaccine development.  相似文献   

17.
Brucella abortus (B. abortus) is a facultative intracellular pathogen that can survive inside macrophages and trophoblast giant cells, and the causative agent of brucellosis. In the present study, we found that production of regulated upon activation normal T-cell expressed and secreted (RANTES) due to B. abortus infection contributes to abortion in pregnant mice. B. abortus infected pregnant interferon-gamma (IFN-gamma) knockout mice died within 15 days of infection, but non-pregnant IFN-gamma knockout mice were still alive. With infection by wild type B. abortus, a large amount of RANTES production was observed in pregnant IFN-gamma knockout mice, and induction of RANTES was also observed in normal pregnant mice infected with the wild type, but not in those infected with the intracellular replication-defective mutant. Production of RANTES and IFN-gamma were inhibited in mice inoculated with the respective RANTES or IFN-gamma antibody. Neutralization of RANTES, induced by B. abortus infection, served to prevent abortion. These results indicate that the production and function of RANTES are correlated with IFN-gamma in pregnant mice infected with B. abortus.  相似文献   

18.
A Neospora-like protozoon found in an aborted bovine placenta   总被引：1，自引：0，他引：1  
Portions of the placenta and kidneys from a 5-month-old bovine fetus were examined histologically. The placental cotyledonary villi were necrotic and protozoa were found in lesions. The organisms were in trophoblasts, periodic acid Schiff-negative and did not react in an immunoperoxidase test using anti-Toxoplasma gondii serum. Individual zoites were approximately 5 x 2 microns and the biggest collection was 45 x 35 microns. The organism in the present case was structurally distinct from Sarcocystis spp. and Toxoplasma gondii, but resembled Neospora caninum.  相似文献   

19.
A subcutaneous extraskeletal giant cell tumor (malignant fibrous histiocytoma) was excised repeatedly from a 9-year-old Domestic Shorthair cat. Ultrastructurally, the mass was composed of fibroblast-like, histiocyte-like, and multinucleated giant cells, and some undifferentiated cells and mononuclear cells intermediate between the fibroblast-like and histiocyte-like cells. Fibroblast-like cells were characterized by abundant well-developed rough endoplasmic reticulum, relatively smooth cytoplasmic membranes, few lysosomal structures, and finely granular chromatin. Histiocyte-like cells resembled immature macrophages. The cell membranes had many villous projections. Rough endoplasmic reticulum varied in quantity. Lysosomes were numerous. Multinucleated giant cells had characteristics of both the fibroblast-like and histiocyte-like cells. No viral particles were seen.  相似文献   

20.
为确定鸡轮状病毒感染的病理变化特点，对自然发病和人工发病轮状病毒感染鸡进行了病理学研究。结果表明，该病特征性的病理剖检变化为肠黏膜脱落出血，呈卡他性炎症表现。特征性组织学变化为小肠绒毛显著缩短，肠腺深度增加；除肠绒毛缩短外，肠上皮还变为扁平或立方形，并伴发坏死、脱落，黏膜充血、水肿与淋巴细胞浸润。电镜观察，肠黏膜上皮细胞的线粒体肿胀，嵴变粗，排列紊乱；粗面内质网及滑面内质网均扩张呈圆形，粗面内质网表面的核糖体部分脱落，胞浆内游离的核糖体减少。  相似文献   

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