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Campylobacter fetus is Internalized by Bovine Endometrial Epithelial Cells

Polish Journal of Microbiology
Volume 68 (2019): Issue 2 (June 2019)
By:
Open Access
|Jun 2019

Figures & Tables

Fig. 1.

Epithelial cells from bovine endometrium maintained in DMEM supplemented with 10% fetal bovine serum.A) Normal appearance of the epithelial-like cells (20×). B) Immunofluorescence. Cytokeratin 18 of epithelial cells of bovine endometrium stained with Alexa 488 (green) (40×). C) The results of RT-PCR for keratin 8 on 2% agarose gel after staining with ethidium bromide. In left lane: DNA ladder (Thermo Fisher Scientific); right lane: the amplicon of keratin 8 (215 pb).
Epithelial cells from bovine endometrium maintained in DMEM supplemented with 10% fetal bovine serum.A) Normal appearance of the epithelial-like cells (20×). B) Immunofluorescence. Cytokeratin 18 of epithelial cells of bovine endometrium stained with Alexa 488 (green) (40×). C) The results of RT-PCR for keratin 8 on 2% agarose gel after staining with ethidium bromide. In left lane: DNA ladder (Thermo Fisher Scientific); right lane: the amplicon of keratin 8 (215 pb).

Fig. 2.

The adherence assay. Epithelial cells from bovine endometrium were challenged with C. fetus (A) or E. coli (B) for 1 h, fixed with methanol and stained with Giemsa (100×). The arrows show the adhered bacteria.
The adherence assay. Epithelial cells from bovine endometrium were challenged with C. fetus (A) or E. coli (B) for 1 h, fixed with methanol and stained with Giemsa (100×). The arrows show the adhered bacteria.

Fig. 3.

The intracellular survival assay. Epithelial cells of endometrium of bovine were infected with C. fetus ATCC 27374 (a MOI of 1000:1). The intracellular bacteria were recovered and plated on the Campylobacter selective agar supplemented with 5% of blood. Average log CFU are shown at 0, 2, 4, 10 and 24 h p.i.
The intracellular survival assay. Epithelial cells of endometrium of bovine were infected with C. fetus ATCC 27374 (a MOI of 1000:1). The intracellular bacteria were recovered and plated on the Campylobacter selective agar supplemented with 5% of blood. Average log CFU are shown at 0, 2, 4, 10 and 24 h p.i.

Fig. 4.

Confocal differential fluorescent staining of internal C. fetus ATCC 27374 on the infected epithelial cells of bovine endometrium. Epithelial cells were grown on coverslips and infected with C. fetus ATCC 27374 at a MOI of 1000:1. Cytoskeleton was stained with phalloidin-FITC (green), and the bacteria with Alexa 594 (red) 2 h p.i. White arrows show intracellular C. fetus (70×).
Confocal differential fluorescent staining of internal C. fetus ATCC 27374 on the infected epithelial cells of bovine endometrium. Epithelial cells were grown on coverslips and infected with C. fetus ATCC 27374 at a MOI of 1000:1. Cytoskeleton was stained with phalloidin-FITC (green), and the bacteria with Alexa 594 (red) 2 h p.i. White arrows show intracellular C. fetus (70×).

Fig. 5.

The cytoskeleton inhibition assay. Epithelial cells of endometrium of bovine were treated with cytocalasin D or nocodazole before and during infection. The cells were infected with C. fetus ATCC 27374 (a MOI of 1000:1). The intracellular bacteria were recovered and plated on the Campylobacter selective agar supplemented with 5% of blood. Average log CFU are shown at 0 and 2 p.i. T-test was performed. All treatments were compared to the not-treatment control, *(p < 0.001).
The cytoskeleton inhibition assay. Epithelial cells of endometrium of bovine were treated with cytocalasin D or nocodazole before and during infection. The cells were infected with C. fetus ATCC 27374 (a MOI of 1000:1). The intracellular bacteria were recovered and plated on the Campylobacter selective agar supplemented with 5% of blood. Average log CFU are shown at 0 and 2 p.i. T-test was performed. All treatments were compared to the not-treatment control, *(p < 0.001).
DOI: https://doi.org/10.33073/pjm-2019-022 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 217 - 224
Submitted on: Dec 5, 2018
Accepted on: Feb 14, 2019
Published on: Jun 28, 2019
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 times per year
Keywords:
bacterial infection,
pathogenicity,
virulence,
pathogen-host interaction,
infectivity
Related subjects:
Life sciences,
Microbiology and virology

© 2019 LIZETH GUADALUPE CAMPOS-MÚZQUIZ, ESTELA TERESITA MÉNDEZ-OLVERA, BEATRIZ ARELLANO-REYNOSO, DANIEL MARTÍNEZ-GÓMEZ, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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