-Q fever is a widespread zoonosis caused by Coxiella burnetii. Infected animals, shedding bacteria by different routes, constitute contamination sources for humans and the environment. To study Coxiella excretion, pregnant goats were inoculated by the subcutaneous route in a site localized just in front of the shoulder at 90 days of gestation with 3 doses of bacteria
Two abortions associated with Coxiella burnetii occurred in a group of 34 pregnant ewes. The seroprevalence of C. burnetii infection was studied by using an ELISA and the immunofluorescence (IF) assay was applied to the contents of vaginal swabs. In addition, a PCR assay, with primers based on a transposon-like repetitive region of the C. burnetii genome (trans-PcR), was used for the highly sensitive and specific detection of C. burnetii in vaginal swabs, milk and faeces. Of the 34 animals tested at parturition, eight (24 per cent) were positive by ELISA, 11 (32 per cent) were positive by IF, and 15 (44 per cent) were positive when the vaginal swab extract was subjected to the trans-PCR assay. C. burnetii was therefore detected by PCR in the vaginal swabs of seven seronegative ewes. However, five weeks after lambing, 16 (47 per cent) of the animals tested were ELISA positive but only two animals (6 per cent) were positive by PCR. Among the ELISA- and PCR-positive animals, eight (25 per cent) shed coxiella in their milk and six (18 per cent) did so in their faeces.
Four chicken lines, L2, B13, PA12 (egg-type), and Y11 (meat-type), were tested for experimental carrier state of Salmonella enteritidis (SE) in two identical trials. After oral inoculation of SE at 1 wk of age with 5 x 10(4) SE colony-forming units (CFU), 10 chickens per line were necropsied weekly for 6 wk and then every 8 or 15 days until the 12th week postinoculation (PI). Liver, spleen, ovary, and ceca were examined for level of SE colonization. Numbers of positive livers and spleens and levels of the challenge strain in these organs differed little between the four chicken lines. Only three positive ovaries were detected. According to the chicken line, ceca exhibited generally significant (P < 0.05) differences in the number of positive organs during weeks 5-11 PI, in the SE CFU levels (P < 0.05) in the first 5 wk PI and during weeks 8 and 10 PI, and in the duration of colonization. L2 and B13 chickens generally carried SE in their ceca at higher levels, in more animals, and for a longer time than PA12 and Y11 chickens. Y11 chickens were the most resistant to SE cecal colonization.
Quantification of the carrier state of Salmonella enteritidis in chicks (i.e., persistent asymptomatic association of S. enteritidis with the host), should provide an optimized means for further investigations into this problem. We therefore developed an experimental carrier state model by oral inoculation of low doses (10(2)-10(4)) of S. enteritidis in B13 chicks at different ages. Liver, spleen, and ceca colonizations by the challenge strains were measured weekly by enumeration of S. enteritidis colony-forming units (CFU) for 7-12 weeks. High mortality rates, incompatible with the carrier state, were observed in chicks inoculated with 10(2) organisms of either a parental strain of S. enteritidis (5556) or a mutant resistant to streptomycin (Smr) and nalidixic acid (Nalr) (strain 1009) at 1 day old. Both strains colonized organs similarly, allowing us to use subsequently the SmrNalr mutant strain. The selected low doses of S. enteritidis induced no deaths in chicks inoculated at 1 or 3 weeks of age. However, inoculation of 3-week-old chicks did not induce a satisfactory carrier state; organ colonization by S. enteritidis was weak and transient, even after inoculation of 10(8) SE. In contrast, some birds infected at 1 week of age presented the challenge strain in the liver and spleen for 3 weeks after inoculation and in the ceca for 12 weeks postchallenge. Most of these birds were colonized by S. enteritidis in the liver and in the ceca for 3 weeks and 10 weeks, respectively, following inoculation. Generally, CFU levels were highest during the first week(s) after inoculation and then decreased progressively. Levels of S. enteritidis were lower in the liver and spleen than in the ceca. Oral inoculation of 1-week-old birds with 5 x 10(4) S. enteritidis provided the required model, allowing quantification of the carrier state of S. enteritidis in chicks.
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