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Campylobacter jejuni is a zoonotic pathogen, and a hypervirulent clone, named clone SA, has recently emerged as the predominant cause of ovine abortion in the United States. To induce abortion, orally ingested Campylobacter must translocate across the intestinal epithelium, spread systemically in the circulation, and reach the fetoplacental tissue. Bacterial factors involved in these steps are not well understood. C. jejuni is known to produce capsular polysaccharide (CPS), but the specific role that CPS plays in systemic infection and particularly abortion in animals remains to be determined. In this study, we evaluated the role of CPS in bacteremia using a mouse model and in abortion using a pregnant guinea pig model following oral challenge. Compared with C. jejuni NCTC 11168 and 81-176, a clone SA isolate (IA3902) resulted in significantly higher bacterial counts and a significantly longer duration of bacteremia in mice. The loss of capsule production via gene-specific mutagenesis in IA3902 led to the complete abolishment of bacteremia in mice and abortion in pregnant guinea pigs, while complementation of capsule expression almost fully restored these phenotypes. The capsule mutant strain was also impaired for survival in guinea pig sera and sheep blood. Sequence-based analyses revealed that clone SA possesses a unique CPS locus with a mosaic structure, which has been stably maintained in all clone SA isolates derived from various hosts and times. These findings establish CPS as a key virulence factor for the induction of systemic infection and abortion in pregnant animals and provide a viable candidate for the development of vaccines against hypervirulent C. jejuni ., (Copyright © 2017 American Society for Microbiology.)

Although Campylobacter jejuni is a common cause of ruminant abortion with great economic impact, the organism has rarely been implicated in canine pregnancy loss, with only 2 documented cases to date. In the current report, 2 cases of perinatal death in adult female Bulldogs associated with C. jejuni infection of fetoplacental organs are described. Fetuses and placentas were received at the Iowa State University Veterinary Diagnostic Laboratory (Ames, Iowa) from 3 puppies that died soon after the birth (case 1) and from an aborted fetus (case 2). Microscopic examination of tissues was generally unremarkable; however, multifocal hemorrhage and infiltrates of macrophages and neutrophils were observed in placental sections from the first case (case 1), and low to moderate numbers of degenerate neutrophils were apparent within multifocal alveoli in the fetal lung in the second case (case 2). Ancillary diagnostics for common infectious causes of reproductive failure in dogs were negative. However, C. jejuni was isolated from the submitted placentas in high numbers in both cases as well as from the fetal lungs and livers. Genotyping of the abortion isolates indicated that the isolates were distinct from each other as well as from selected canine enteric C. jejuni isolates included herein for comparison. Both abortion strains were sensitive to all 9 antimicrobials tested, except the isolate from case 2, which displayed resistance to tetracycline. These findings provide convincing evidence for the inclusion of C. jejuni culture in routine diagnostic testing for causes of canine pregnancy loss., (© 2014 The Author(s).)

The pregnant guinea pig is an effective model for studying abortifacient Campylobacter spp, and previous experiments have demonstrated that C. jejuni IA3902 has a marked predilection for the subplacenta while sparing the placental disc in this species. In the study described here, the growth and chemotaxis of IA3902 and a reference strain (NCTC 11168) are compared in the presence of subplacental and placental factors, as well as bile and plasma, from pregnant and nonpregnant guinea pigs. Both strains grew better in subplacental versus placental disc tissue extracts at 24 hours; however, only IA3902 maintained this enhancement at 48 hours. Histochemistry and lectin histochemistry were used to localize mucin, iron, and l-fucose within the placental unit. Mucin was most abundant in subplacental lacunae, the junctional zone, and visceral yolk sac placenta, while iron was most abundant in the placental disc, and L-fucose-containing surface glycans were limited to the visceral yolk sac placenta. These 3 individual factors, along with progesterone and estradiol, were evaluated for effects on growth and chemotaxis of C. jejuni. Mucin, iron, and L-fucose were growth promoting, while l-fucose was also chemoattractive for both strains. Progesterone, estradiol, and pregnant guinea pig plasma did not affect growth or chemotaxis, and no difference was observed when bile from pregnant and nonpregnant animals was compared. These findings demonstrate the presence of specific factors within the guinea pig placental unit that drive chemotaxis and enhance growth of C. jejuni, shedding light on potential mechanisms underlying the fetoplacental tropism observed with this strain.

Objective: To compare efficacy of 2 commercial ovine Campylobacter vaccines and an experimental bacterin in guinea pigs following IP inoculation with Campylobacter jejuni IA3902., Animals: 51 female guinea pigs., Procedures: Pregnant and nonpregnant animals were randomly assigned to 1 of 4 treatment groups and administered a commercial Campylobacter vaccine labeled for prevention of campylobacteriosis in sheep via two 5-mL doses 14 days apart (vaccine A; n = 13), another labeled for prevention of campylobacteriosis via two 2-mL doses (vaccine B; 12), an experimental bacterin prepared from the challenge strain (12), or a sham vaccine (14). Ten days later, animals were challenged IP with C jejuni IA3902; 48 hours later, animals were euthanized, complete necropsy was performed, and blood and tissue samples were obtained for bacteriologic culture., Results: Administration of vaccine B or the experimental bacterin, but not vaccine A, significantly reduced 48-hour infection rates versus administration of the sham vaccine. A significantly reduced 48-hour infection rate was associated with administration of vaccine B independent of pregnancy status., Conclusions and Clinical Relevance: Administration of vaccine B significantly reduced infection in guinea pigs challenged with C jejuni IA3902, similar to a homologous bacterin. Results suggested that vaccine B or an autogenous product may be effective in controlling ovine campylobacteriosis caused by this emergent abortifacient strain. Bacteriologic culture of blood, liver, bile, and uterus in nonpregnant guinea pigs 48 hours after inoculation may be a useful screening tool for comparing efficacy of C jejuni vaccines.

Toll-like receptors 2 and 4 (TLR2 and TLR4) are well-characterized cell surface receptors that recognize specific pathogen-associated molecular patterns and play an important role in pathogen recognition and activation of the innate immune system. Variable expression of TLR2 and TLR4 has been described in trophoblasts from normal and diseased placentas; yet, there are limited data regarding trophoblast TLR expression in response to specific placental pathogens, and TLR expression in the guinea pig placenta has not been described. The guinea pig is an effective model for Campylobacter-induced abortion of small ruminants, and the authors have shown by immunohistochemistry that C jejuni localizes within syncytiotrophoblasts of the guinea pig subplacenta. The present study was designed to determine if the expression of either TLR2 or TLR4 would be affected in subplacental trophoblasts following infection with C jejuni. Immunohistochemistry for TLR2 and TLR4 was performed on placenta from guinea pigs that aborted following inoculation with C jejuni and from sham-inoculated controls. Quantitative assessment of TLR expression was performed, and mean immunoreactivity for TLR2 was significantly higher in subplacental trophoblasts from animals that aborted compared with uninfected controls (P = .0283), whereas TLR4 expression was not statistically different (P = .5909). These results suggest that abortion in guinea pigs following infection with C jejuni is associated with increased TLR2 expression in subplacental trophoblasts and may reveal a possible role for TLR2 in the pathogenesis of Campylobacter-induced abortion.

Objective: To compare pathogenicity of an emergent abortifacient Campylobacter jejuni (IA 3902) with that of reference strains after oral inoculation in pregnant guinea pigs., Animals: 58 pregnant guinea pigs., Procedures: 12 animals were challenged IP with C jejuni IA 3902 along with 5 sham-inoculated control animals to confirm abortifacient potential. Once pathogenicity was confirmed, challenge via oral inoculation was performed whereby 12 guinea pigs received IA 3902, 12 received C jejuni isolated from ovine feces (OF48), 12 received a fully sequenced human C jejuni isolate (NCTC 11168), and 5 were sham-inoculated control animals. After abortions, guinea pigs were euthanized; samples were collected for microbial culture, histologic examination, and immunohistochemical analysis., Results: C jejuni IA 3902 induced abortion in all 12 animals following IP inoculation and 6 of 10 animals challenged orally. All 3 isolates colonized the intestines after oral inoculation, but only IA 3902 induced abortion. Evidence of infection existed for both IA 3902 and NCTC 11168; however, C jejuni was only recovered from fetoplacental units of animals inoculated with IA 3902. Immunohistochemical analysis localized C jejuni IA 3902 infection to subplacental trophoblasts, perivascular tissues, and phagocytes in the placental transitional zone., Conclusions and Clinical Relevance: This study revealed that C jejuni IA 3902 was a unique, highly abortifacient strain with the ability to colonize the intestines, induce systemic infection, and cause abortion because of its affinity for the fetoplacental unit. Guinea pigs could be effectively used in the study of septic abortion after oral inoculation with this Campylobacter strain.