scholarly journals Investigating the Suitability of a Laboratory Mouse Model to Study the Pathogenesis of Abortifacient Campylobacter jejuni

2019 ◽  
Vol 56 (4) ◽  
pp. 555-564
Author(s):  
Victoria D. Lashley ◽  
Michael J. Yaeger ◽  
Ju Ji ◽  
Orhan Sahin ◽  
Zuowei Wu
Keyword(s):  
Campylobacter Jejuni ◽  
Reproductive Tract ◽  
Laboratory Mouse ◽  
Intrauterine Fetal Death ◽  
Microbial Culture ◽  
Large Numbers ◽  
Colony Forming Units ◽  
Pregnant Mice ◽  
Neutral Mucin ◽  
Reproductive Pathology

The aim of this study was to assess whether pregnant mice represent a useful model to study the reproductive pathology of Campylobacter jejuni IA3902 using the end point of positive microbial culture of the organism from the fetoplacental unit. Pregnant BALB/c and CD-1 mice (14 days’ gestation) were inoculated orally and intraperitoneally (IP) with 1 × 109 colony-forming units/ml of C. jejuni IA3902. The organism was recovered by microbial culture from the fetoplacental unit in 10 of 10 BALB/c and 10 of 10 CD-1 IP-inoculated pregnant mice and in 29% (2/7) BALB/c and 38% (3/8) CD-1 orally inoculated pregnant mice. Gross reproductive lesions included necrosuppurative placentitis, fetal resorption, intrauterine fetal death, stillborn pups (dead neonates), and multifocal hepatitis. Histological changes consisted of locally extensive neutrophilic and necrotizing placentitis with intralesional bacterial colonies of C. jejuni, ulcerative endometritis, random multifocal hepatitis, and rare cholecystitis. Immunohistochemistry for the major outer membrane protein of C. jejuni revealed moderate to large numbers of the organism at the periphery of the placental discs, within trophoblasts and extracellularly, with invasion into the placental disc largely via the vascular network. The organism is trophic for neutral mucin, iron, and L-fucose within the murine placenta. C. jejuni IA3902 has affinity for the murine reproductive tract, specifically the fetoplacental unit, where it results in a necrotizing placentitis with positive microbial recovery after both IP and oral challenge in BALB/c and CD-1 pregnant mice.

201. Effects of relaxin deficiency on matrix metalloproteinase expression in the cervix and vagina of pregnant mice

2005 ◽  
Vol 17 (9) ◽  
pp. 76
Author(s):  
J. T. McGuane ◽  
H. M. Gehring ◽  
L. J. Parry
Keyword(s):  
Reproductive Tract ◽  
Late Gestation ◽  
Mrna Levels ◽  
Biochemical Processes ◽  
Fluorogenic Probes ◽  
Pregnant Mice ◽  
Ecm Degradation ◽  
Stromal Collagen

The major functions of relaxin are associated with female reproductive physiology, especially the regulation of biochemical processes involved in the remodelling of the reproductive tract at term. Studies in relaxin deficient mice (Rlx–/–) demonstrate that although females give birth to live young without apparent dystocia, they have abnormal cervices and vaginae. This phenotype is attributed to an increase in stromal collagen, but the mechanism(s) by which relaxin regulates extracellular matrix (ECM) production in reproductive tissues is poorly understood. In this study, we assessed the expression of matrix metalloproteinases (MMPs) in the cervix and vagina of pregnant wild-type (Rlx+/+) and Rlx–/– mice. Tissues were obtained from adult C57/Blk6J Rlx+/+ mice on days 7.5, 14.5, 17.5, 18.5 pc and Rlx–/– littermates on days 7.5, 14.5 and 18.5 pc. Real-time PCR using dual-labelled fluorogenic probes was performed in an Opticon 2 cycler (MJ Research) to quantify MMP-2, -3, -7, -9 and -13 gene expression. In the cervix and vagina of Rlx+/+ mice, only MMP-2 mRNA levels were significantly higher at term compared with earlier stages of gestation. There were significant decreases in MMP-7 and -13 expression at term, but no change in MMP-3 and -9. In contrast, MMP-3, -7, -9 and -13 mRNA levels were significantly higher in the cervix and vagina of late pregnant Rlx–/– mice. The expression of MMP-2 did not differ between Rlx+/+ and Rlx–/– mice at term. Despite the higher expression of the majority of MMPs we examined in Rlx–/– mice, there was no histological evidence of increased ECM degradation in the cervix and vagina in late gestation. Although previous in vitro studies suggest that relaxin positively regulates MMP activity, our data demonstrate that relaxin deficiency does not result in decreased MMP expression in the mouse cervix and vagina in vivo.

scholarly journals Potentials of Mouthwashes in Disinfecting Cariogenic Bacteria and Biofilms Leading to Inhibition of Caries

2012 ◽  
Vol 6 (1) ◽  
pp. 23-30 ◽  
Author(s):  
Takehiro Oyanagi ◽  
Junji Tagami ◽  
Khairul Matin
Keyword(s):  
Ex Vivo ◽  
Secondary Caries ◽  
Bacterial Viability ◽  
Cariogenic Bacteria ◽  
Benzethonium Chloride ◽  
Large Numbers ◽  
Carious Lesions ◽  
Colony Forming Units ◽  
Caries Model ◽  
Phosphate Buffered Saline

Objectives:The aim of this study was to compare the effects of certain commercially available mouthwashes on cariogenic bacteria and biofilms, following the acquisition of inhibition potentials of caries.Materials and Methods:Mouthwashes containing I) chlorhexidine gluconate (CHG; 0.0005% w/v), II) benzethonium chloride (BTC; 0.01% w/v), III) an essential oil (Listerine), and IV) povidone-iodine (PVP-I; 0.035% w/v) were tested on planktonic cariogenic bacteria, biofilms, and an ex vivo caries model. Bacterial aliquots were inoculated with each solution separately and vortexed for 10 seconds at room temperature. Bacterial viability was subsequently investigated by fluorescence microscopy (FM) after staining with a BacLight viability kit and the number of colony-forming units (CFUs) was counted. Similarly, mouthwash solutions were applied to artificial cariogenic biofilms, and bacterial viability of the biofilms was investigated as stated above. Inhibition potentials of two selected mouthwashes of carious lesions were investigated using biofilm-induced caries and a secondary caries model. In all steps, a phosphate-buffered saline (PBS) solution was included as a control.Results:Planktonic cariogenic bacteria and bacteria embedded in biofilms were killed in remarkably large numbers with Listerine and PVP-I treatment compared to PBS and other gargles. CFU counts also showed significant reduction after treatment with Listerine and PVP-I compared to other solutions (P<0.05). Listerine also displayed significant (P<0.05) inhibition effects in preventing the progression of demineralization.Conclusion:Bactericidal potencies of the mouthwashes varied significantly, suggesting that mouthwashes like Listerine can be useful for the prevention of caries and secondary caries.

Survival of Campylobacter jejuni in Fresh and Heated Red Meat

1983 ◽  
Vol 46 (9) ◽  
pp. 771-774 ◽  
Author(s):  
PAUL KOIDIS ◽  
MICHAEL P. DOYLE
Keyword(s):  
Heat Treatment ◽  
Campylobacter Jejuni ◽  
Ground Beef ◽  
Red Meat ◽  
Refrigerated Storage ◽  
Log10 Reduction ◽  
Salmonella Spp ◽  
Lamb Meat ◽  
Large Numbers ◽  
D Values

Studies were done to assess the ability of Campylobacter jejuni to survive in fresh ground beef during refrigerated storage and to identify time-temperature treatments needed to inactivate Campylobacter in ground and cubed red meat. The organism survived well in refrigerated ground beef containing large numbers of indigenous bacteria. Relatively little death (&lt; 1.2-log10 reduction) occurred for 7 of 8 strains during 14 d at 4°C. C. jejuni inoculated into ground beef and cubed lamb meat was quite sensitive to heat treatment. D-values for inactivation of campylobacters in ground beef ranged from 5.9 to 6.3 min at 50°C and from 12 to 21 s at 58°C. D-values were generally greater when campylobacters were heated in lamb meat, ranging from 5.9 to 13.3 min and 12.5 to 15.8 s at 50 and 60°C, respectively. All strains of C. jejuni were more sensitive to heat than salmonellae, hence meat heated to a temperature sufficient to inactivate Salmonella spp. should be free of viable campylobacters.

scholarly journals Collection of pluripotential hematopoietic stem cells by cytapheresis

Blood ◽  
1982 ◽  
Vol 59 (4) ◽  
pp. 822-827 ◽  
Author(s):  
LC Lasky ◽  
RC Ash ◽  
JH Kersey ◽  
ED Zanjani ◽  
J McCullough
Keyword(s):  
Stem Cells ◽  
Collection Efficiency ◽  
Hematopoietic Stem ◽  
Hematopoietic Reconstitution ◽  
Large Numbers ◽  
Colony Forming Units ◽  
Peripheral Stem Cells ◽  
And Storage ◽  
Marrow Ablation ◽  
Cell Colony

Abstract Successful complete hematopoietic reconstitution (CHR) using nonleukemic peripheral stem cells (PSC) after marrow ablation has been reported in animals but not man. Previous studies of cytapheresis products from humans, as a prelude to use for CHR, have documented the presence of committed myeloid (CFU-GM) and erythroid (BFU-E) precursors. We have examined mononuclear cell (MNC) products collected on the Fenwal CS3000 Blood Cell Separator for these plus the more primitive mixed (granulo-, erythro-, mono-, and megakaryocytic) cell colony-forming units (CFU-GEMM) and for various lymphocytic subpopulations (LSP). One to two-hour products contained 36 +/- 7 CFU- GEMM/10(6) MNC (mean +/- SE, n = 8) or 490 +/- 131/ml product. This compared favorably with blood (23 +/- .4/10(6) MNC or 46 +/- 8/ml, n = 14) and bone marrow (146 +/- 58/10(6) MNC, n = 12). Collection efficiency for E-rosette-positive cells approximated that for total lymphocytes and was variable for other LSP. Recovery of CFU-GEMM after freezing in 10% dimethylsulfoxide at a controlled rate and storage in liquid N2 was 54% +/- 8% (n = 8). Cytapheresis collection of large numbers of pluripotent hematopoietic precursors and demonstration of adequate recovery of these after cryopreservation, both previously unreported, are significant steps toward eventual CHR using nonleukemic PSC.

THE EFFECTS OF OESTRADIOL-3,17β ON TUBAL TRANSPORT IN THE LABORATORY MOUSE

1968 ◽  
Vol 42 (1) ◽  
pp. 17-26 ◽  
Author(s):  
K. W. HUMPHREY
Keyword(s):  
Laboratory Mouse ◽  
Concurrent Administration ◽  
Oestradiol Treatment ◽  
Pregnant Mice ◽  
Delayed Development

SUMMARY Pregnant mice received varying doses of oestradiol on days 1–3 of pregnancy, or on day 1 or day 2 only. The effects of oestradiol on tubal transport depend largely on the time of treatment rather than the dose of oestradiol. Treatment with 1·6 μg. on day 1 when the ova were still in the ampulla caused retention of all ova in the oviduct until day 4—many of these tube-locked ova were retained in the ampulla, apparently because of prolonged closure of the ampulla-isthmus junction. Treatment with 0·4 μg. on day 2 when all ova were in the isthmus caused premature entry of ova into the uteri and reduced the recovery of ova by over 50%. Ligation of various parts of the tract and ovum transfer studies showed that this loss of ova is due to accelerated transport from the isthmus to the uterus and vagina. These effects on tubal transport were not reversed by concurrent administration of progesterone. Tubal retention was associated with delayed development of the blastocoele but these ova implanted normally after ovum transfers.

' Bacillus piliformis ' (Tyzzer) and Tyzzer’s disease of the laboratory mouse I. Propagation of the organism in embryonated eggs

1966 ◽  
Vol 165 (998) ◽  
pp. 35-60 ◽  
Keyword(s):  
Epithelial Cells ◽  
Laboratory Mouse ◽  
Serial Passage ◽  
Yolk Sac ◽  
Liver Lesions ◽  
Hepatic Cells ◽  
Large Numbers ◽  
Significant Difference ◽  
Hepatic Lesions

A contagious disease of laboratory mice characterized by focal necrotic lesions of the liver was first described by Tyzzer (1917). This author described a pleomorphic sporing organism associated with the disease and called it ' Bacillus piliformis '. He found it intracellularly in hepatic cells bordering the necrotic foci and also in epithelial cells of the large intestine of mice showing liver lesions. Many cells contained large numbers of organisms, and Tyzzer concluded that multiplication was intracellular. B . piliformis has not been cultivated on bacteriological media and hitherto the only sources available for experimental study of Tyzzer’s disease have been infected mouse liver and brain. With such material it is difficult to produce liver lesions in mice by parenteral injection unless the animals are treated with cortisone. This communication describes the isolation and serial passage of two strains of B . piliformis in embryonated eggs. One strain was propagated for 30 serial passages, 26 of which were made in eggs by yolk sac injection and the others in mice. A non-sporing variant obtained from this strain produced hepatic lesions in embryos and mice similar to those produced by the parent sporing strain. No evidence was obtained of extracellular growth of the organism and intracellular growth occurred almost exclusively in epithelial cells of the yolk sac endoderm and hepatic cells of the embryo. Although pleomorphic, B . piliformis has a distinctive morphology which was described in detail by Tyzzer. All the forms described by this author were observed in embryonated eggs. The vegetating form of the organism rapidly lost its infectivity in vitro and also in the egg following death of the embryo. This loss of infectivity proved a limitation to certain in vitro studies and no method of halting it in the fluid state was discovered. However, it proved possible to obtain sufficient survival of the non-sporing strain for inoculation purposes by preserving crude tissue suspensions at – 75 °C. The spores survived heating at 56 °C but were usually inactivated at 65 °C. They survived well at room temperature and the organism was recovered by mouse and egg passage from inoculated eggs kept at this temperature for over a year. Penicillin was found to be highly effective in halting the progress of infection in eggs. In the course of the work there were indications of a cyclic variation in the susceptibility of embryonated eggs to infection, and on one occasion a significant difference was demonstrable between eggs obtained from different pens. The results obtained when yolk sac and chick embryo liver infected with B . piliformis were injected into mice are described in the following paper (Craigie 1966).

Sensitivity of Campylobacter jejuni to Drying

1982 ◽  
Vol 45 (6) ◽  
pp. 507-510 ◽  
Author(s):  
MICHAEL P. DOYLE ◽  
DEBRA J. ROMAN
Keyword(s):  
Campylobacter Jejuni ◽  
Room Temperature ◽  
Skim Milk ◽  
Initial Population ◽  
Log10 Reduction ◽  
Viable Cells ◽  
Large Numbers ◽  
Thermophilic Campylobacter ◽  
And Storage ◽  
Intermediate Rate

Several factors were shown to influence the rate of inactivation of Campylobacter sp. when dried on a glass surface. These included strain, temperature and humidity, and medium used to suspend the organism. Of the strains evaluated, all of three isolates of Campylobacter jejuni exhibited greater tolerance to drying than did a strain of nalidixic acid resistant, thermophilic Campylobacter. Inconsistent results were obtained when organisms were dried and maintained at 25 C. Viable cells from two of four strains having an initial population of &gt;107 were not recovered after 24 h in an anhydrous environment at 25 C. Under comparable conditions, drying C. jejuni FRI-CF8 in the presence of skim milk at 25 C resulted in a &gt;107 log10 reduction of cells within 1 day in one instance; a 5 log10 decline after 7 days in another; and inactivation at an intermediate rate on a third occasion. Rates of death were greatly reduced when cells were dried and held at 4 C. At this temperature and in the presence of skim milk and an anhydrous environment, a 5 log10 reduction of CF8 occurred after 6 weeks. In all instances, greater survival occurred when organisms were dried in the presence of Brucella broth than in skim milk. When held in environments of different relative humidities (RH), survival was greatest in the presence of 14% or less RH. Results suggest that C. jejuni is generally quite sensitive to drying and storage at room temperature, but, at refrigeration temperature and the appropriate humidity, large numbers may survive drying and remain viable for several weeks.

Improvement of capture efficacy of immunomagnetic beads forCampylobacter jejuniusing reagents that alter its motility

2013 ◽  
Vol 59 (7) ◽  
pp. 511-514 ◽  
Author(s):  
Hongsheng Huang ◽  
Beverley Phipps-Todd
Keyword(s):  
Campylobacter Jejuni ◽  
Contact Time ◽  
Detection Limits ◽  
Immunological Reactivity ◽  
Immunomagnetic Beads ◽  
Chain Reaction ◽  
Slowing Down ◽  
Colony Forming Units ◽  
High Detection ◽  
Polymerase Chain

Previous studies using the immunomagnetic beads separation (IMS) technique have shown high detection limits of live campylobacters but low detection limits of formalin-killed campylobacters. The present study investigated if the addition of various concentrations of reagents that alter the motility of live Campylobacter jejuni could enhance the recovery of the organisms by IMS. The addition of 5% glycerol, 0.001% formalin, 10% polyethylene glycol, or 0.001% agarose in a buffer slowed down the movement of C. jejuni and increased the recovery of live C. jejuni, using beads coated with specific monoclonal antibodies (mAbs). The highest recovery yielded was 5.2- ± 3.3-fold with 5% glycerol at 105colony-forming units (CFU)·mL−1. The addition of 5% glycerol also improved isolation at lower concentrations of C. jejuni (102to 104CFU·mL−1) in buffer. The recovery by IMS of C. jejuni killed by 1% formalin was increased up to as high as 17-fold compared with the recovery of live organisms, as detected using a real-time polymerase chain reaction assay. The reagents investigated did not enhance the immunological reactivity of the mAbs to this organism. These results indicate that the addition of several reagents enhanced the capture of C. jejuni by IMS, which could be partially due to the slowing down of the movement or the altering of the motility of C. jejuni and to the increasing of the contact time between C. jejuni and immunomagnetic beads.

Campylobacter jejuni 81-176 forms distinct microcolonies on in vitro-infected human small intestinal tissue prior to biofilm formation

Microbiology ◽  
2010 ◽  
Vol 156 (10) ◽  
pp. 3079-3084 ◽  
Author(s):  
Graham Haddock ◽  
Margaret Mullin ◽  
Amanda MacCallum ◽  
Aileen Sherry ◽  
Laurence Tetley ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Biofilm Formation ◽  
Target Tissue ◽  
Apical Surface ◽  
Intestinal Tissue ◽  
Time Points ◽  
Transmission Electron ◽  
Large Numbers ◽  
Ileal Tissue

Human small and large intestinal tissue was used to study the interaction of Campylobacter jejuni with its target tissue. The strain used for the study was 81-176 (+pVir). Tissue was processed for scanning and transmission electron microscopy, and by immunohistochemistry for light microscopy. Organisms adhered to the apical surface of ileal tissues at all time points in large numbers, in areas where mucus was present and in distinct groups. Microcolony formation was evident at 1–2 h, with bacteria adhering to mucus on the tissue surface and to each other by flagellar interaction. At later time points (3–4 h), biofilm formation on ileal tissue was evident. Flagellar mutants did not form microcolonies or biofilms in tissue. Few organisms were observed in colonic tissue, with organisms present but not as abundant as in the ileal tissue. This study shows that C. jejuni 81-176 can form microcolonies and biofilms on human intestinal tissue and that this may be an essential step in its ability to cause diarrhoea in man.

scholarly journals Antibiofilm Effects of Heated Scallop Shell Powder on Campylobacter jejuni Biofilms

Membranes ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 43
Author(s):  
Haruka Tsukuda ◽  
Taiki Akimoto ◽  
Nona Fukikoshi ◽  
Resei Wada ◽  
Jun Sawai
Keyword(s):  
Campylobacter Jejuni ◽  
Antibiofilm Activity ◽  
Scallop Shell ◽  
Predominant Species ◽  
Large Numbers ◽  
Scallop Shells ◽  
Naoh Treatment ◽  
Microaerobic Conditions ◽  
Shell Powder ◽  
Bactericidal Agent

Methods to reuse large numbers of scallop shells from the harvesting regions of Japan are being explored. The major component of scallop shells is calcium carbonate (CaCO3), which forms the powerful bactericidal agent, calcium oxide (CaO), when heated. Heated scallop shell powder (HSSP) exhibits strong and broad-spectrum antimicrobial activity against bacteria, fungi, and viruses. This study investigated the antibiofilm activity of HSSP against the biofilms of Campylobacter jejuni, which is the predominant species in campylobacteriosis. Biofilm samples of C. jejuni were prepared on 0.45 µm filter paper under microaerobic conditions. The HSSP treatment inactivated and eradicated C. jejuni biofilms. The resistance of C. jejuni biofilms to HSSP was significantly higher than that of the floating cells. Moreover, the antibiofilm activity of the HSSP treatment against C. jejuni biofilms was higher than that of NaOH treatment at the same pH. These results indicated that HSSP treatment is an effective method for controlling C. jejuni biofilms.

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