Microbiota attenuates chicken transmission-exacerbated campylobacteriosis in Il10−/− mice

Abstract

Campylobacter jejuni is a prevalent foodborne pathogen mainly transmitting through poultry. It remains unknown how chicken-transmitted C. jejuni and microbiota impact on human campylobacteriosis. Campylobacter jejuni AR101 (Cj-P0) was introduced to chickens and isolated as passage 1 (Cj-P1). Campylobacter jejuni Cj-P1-DCA-Anaero was isolated from Cj-P0-infected birds transplanted with DCA-modulated anaerobic microbiota. Specific pathogen free Il10−/− mice were gavaged with antibiotic clindamycin and then infected with Cj-P0, Cj-P1, or Cj-P1-DCA-Anaero, respectively. After 8 days post infection, Il10−/− mice infected with Cj-P1 demonstrated severe morbidity and bloody diarrhea and the experiment had to be terminated. Cj-P1 induced more severe histopathology compared to Cj-P0, suggesting that chicken transmission increased C. jejuni virulence. Importantly, mice infected with Cj-P1-DCA-Anaero showed attenuation of intestinal inflammation compared to Cj-P1. At the cellular level, Cj-P1 induced more C. jejuni invasion and neutrophil infiltration into the Il10−/− mouse colon tissue compared to Cj-P0, which was attenuated with Cj-P1-DCA-Anaero. At the molecular level, Cj-P1 induced elevated inflammatory mediator mRNA accumulation of Il17a, Il1β, and Cxcl1 in the colon compared to Cj-P0, while Cj-P1-DCA-Anaero showed reduction of the inflammatory gene expression. In conclusion, our data suggest that DCA-modulated anaerobes attenuate chicken-transmitted campylobacteriosis in mice and it is important to control the elevation of C. jejuni virulence during chicken transmission process.

Recombinant Bile Salt Hydrolase Enhances the Inhibition Efficiency of Taurodeoxycholic Acid against Clostridium perfringens Virulence

“…After 4 h, the cells were collected, and total RNA was extracted using TRIzol (Thermo Fisher Scientific, St. Louis, MO, USA) as described earlier [ 4 , 30 ]. After cDNA was synthesized using M-MLV (NE Biolab, Ipswich, MA, USA) and a random hexamer (NE Biolab, Ipswich, MA, USA) [ 31 ], the virulence gene expression of asrA1 , netB , colA , and virT was determined using SYBR Green PCR Master Mix (Bio-Rad, Hercules, CA, USA) in a Bio-Rad 384-well real-time PCR (qPCR) system (Bio-Rad, Hercules, CA, USA) as described in the previous paper [ 28 ]. The primer sequences of gyrA , asrA1 , and virT genes were described previously [ 28 ] and here were the primer sequences: netB _forward: 5′-ggaaaaatgaaatggcctga-3′; netB _reverse: 5′-gcaccagcagtttttccttc-3′; colA _forward: 5′-taggaacaaaggcgcaagat-3′; colA _reverse: 5′-gaatactgcattccccttgc-3′.…”

“…The primer sequences of gyrA, asrA1, and virT genes were described previously [28] and here were the primer sequences: netB_forward: 5 ′ -ggaaaaatgaaatggcctga-3 ′ ; netB_reverse: 5 ′ -gcaccagcagtttttccttc-3 ′ ; colA_forward: 5 ′ -taggaacaaaggcgcaagat-3 ′ ; colA_reverse: 5 ′ -gaatactgcattccccttgc-3 ′ . The gene expression of the fold-change was calculated using the ∆∆Ct method [31] and gyrA as an internal control.…”

The Role of Immune Response and Microbiota on Campylobacteriosis

“…FMT of mouse anaerobic microbiota to germ free Il10 −/− mice prevents C. jejuni-induced intestinal inflammation with reduced inflammatory genes of Cxcl2, Il17 and Il1β as well as massive immune cell infiltration into gut lamina propria [85]. DCA-modulated anaerobes could attenuate chicken transmission exacerbated campylobacteriosis in mice by reduction of inflammatory genes, Il1 7a, Il1β, and Cxcl1 expression in cellular level and inhibiting mTOR signaling pathway [128].…”

Vaccines Using Clostridium perfringens Sporulation Proteins Reduce Necrotic Enteritis in Chickens

“…The samples were immediately cooled on ice and stored at −80 • C before RNA isolation. RNA was isolated as reported before [42]; briefly, the stored samples suspended in Trizol were thawed, and 200 µL of chloroform was added per 1 mL of Trizol Reagent. The mixture was vortexed thoroughly for 30 s, sat at room temperature for 10 min, and centrifuged at 12,500 rpm for 14 min at 4 • C. The top 500 µL colorless aqueous phase was then transferred to a fresh 1.5 mL microcentrifuge tube.…”

“…mRNA levels of proinflammatory genes of mouse Il1β, Tnfα, and Cxcl2, and chicken Ifnγ, Il1β, and Il8-1, were determined using the SYBR Green PCR Master mix (Bio-Rad Hercules, CA, USA) on a Bio-Rad 384-well Real-Time PCR System and normalized to the respective Gapdh. The primer sequences were reported before [10,14,42] and are in Supplementary Table S1. The PCR reactions were performed according to the manufacturer's recommendation.…”