Experimental Brucella ovis infection in pregnant ewes.

Forty yearling Brucella-free ewes were inoculated with Brucella ovis by the conjunctival route in mid or late first pregnancy. Only a few ewes excreted B ovis during pregnancy and gave birth to stillborn lambs, but most of them excreted the organism at lambing or during lactation. One of the 11 lambs which were born alive but died before they were weaned was found to be infected postmortem. In contrast, none of the 46 surviving lambs which were reared in isolation until adulthood, was found to be infected. At weaning, the 40 ewes were mated again with five Brucella-free rams. Although many of the ewes excreted B ovis, none of the rams was found to be infected when necropsied after mating. Most of the ewes that became pregnant, all having excreted B ovis during their first pregnancy, cleared the infection during the second pregnancy. However, three remained persistently infected and excreted B ovis in their milk throughout the second lactation. None of the lambs born to these three ewes was found to be infected when necropsied at weaning.     

Experimental mixed infection of rabbits with Yersinia enterocolitica and Listeria monocytogenes

Experimental mixed infection was reproduced in rabbits after per os infection with Yersinia enterocolitica serotype 0:3 cells. Four days later some of animals were re-infected orally with Listeria monocytogenes serotype 4b cells. A third group of healthy rabbits was also infected per os with Listeria monocytogenes. The infectious process was followed dynamically from days 1-28. The experimental animals were examined for clinical, paraclinical and morphological findings. Augmentation of body temperature and alveolar macrophage number, a decreased number of peritoneal macrophages, leucopenia as well as purulent meningoencephalitis, catarrhal pneumonia, lienitis, lymphadenitis and enteritis were detected after experimental mixed infection. Both types of macrophages demonstrated a weak bactericidal activity against Yersinia enterocolitica and a highly expressed killing effect against Listeria monocytogenes. Yersinia and Listeria cells were isolated from the viscera and brain. Both species of bacteria were established intracellularly in the macrophages by electron-microscopic examination. The data received showed that mixed Yersinia enterocolitica 0:3 and Listeria monocytogenes 4b infection of rabbits runs with transitory hyperthermia as a generalized infection and is similar to the Listeria mono-infection. The immunosuppressive effect induced by oral Yersinia enterocolitica infection of rabbits promotes the expression of listerious agents.     

Experimental infection of pregnant ewes with enteric and abortion-source Chlamydophila abortus

Two groups of pregnant ewes were experimentally infected oronasally in midpregnancy. A faecal and an abortion-source isolate of Chlamydophila abortus were used. They were derived from a healthy ewe from a flock with no history of abortion, and from an aborted foetus in a farm with enzootic abortion. As assessed by modified Ziehl-Neelsen (MZN) staining, egg culture, antigen ELISA, the Clearview test and immunohistochemistry, inoculation resulted in placental and/or foetal infection in all ewes. Histopathology revealed placentitis in two and four ewes inoculated with the enteric or abortion-source isolate, respectively, in addition, these samples were immunohistochemically positive for chlamydial antigen. All six ewes infected with the enteric isolate and five of seven ewes infected with the abortion-source isolate showed evidence for a serological response by an indirect ELISA or CFT. Neither chlamydiae nor lesions were detected in the placentae and lambs of the uninfected control ewes, which remained seronegative. Our results suggest that enteric C. abortus can be associated with placental and foetal lesions in sheep.     

Experimental intrauterine inoculation of pregnant ewes with ureaplasmas

The pregnancies of 13 ewes which were inoculated intrauterine with one of two strains of ovine ureaplasmas resulted in 9 normal and 3 abnormal births, and one ewe was found to be no longer pregnant on postmortem examination. Vaginal ureaplasma infection was detected in the majority of ewes only after lambing. Of the 12 ewes examined at postmortem, ureaplasmas were isolated from the uterus of 5 out of the 6 necropsied up to 21 days post-partum. The vulvar/preputial areas of the majority of lambs that survived were infected with ureaplasmas for the duration of the experiment, but infections of the nasal cavity and eye areas, detected at birth in 4 lambs, were resolved within 8 days post-partum.The only pathological effects detected that could possibly be attributed to ureaplasma infection were a placentitis in an ewe that aborted, and the resorption of the foetus in another.     

Respiratory infection of turkeys with Listeria monocytogenes Scott A

The pathogenesis of L. monocytogenes strain Scott A was studied by challenging day-old male turkey poults by air sac inoculation with tryptose phosphate broth containing 10(0) cfu (control), 10(4), 10(5), and 10(6) cfu (low challenge), or 10(7) and 10(8) cfu (high challenge) of the Scott A (serotype 4b) strain of L. monocytogenes. Mortality at 2 wk postinfection (PI) ranged from 25% for low challenge to 100% for high challenge (P= 0.0001). Gross and histopathological lesions were observed in heart, liver, spleen, lung, and bursa of Fabricius of mortalities at 4 days PI. Listeria monocytogenes challenge resulted in significantly decreased relative weight of the bursa of Fabricius and increased relative weight of the spleen, and L. monocytogenes was isolated by direct plating of liver, pericardium, brain, and both left and right stifle joint synovium (knee) cultures, as well as gall bladder, yolk sac, and cecal tonsil from transfer swabs onto Listeria-selective agar. Isolates were confirmed as positive using Gram stain, biochemical tests, and the Biolog system. High challenge resulted in confirmed L. monocytogenes isolation from 48% of left knee and 59% of right knee cultures. Low challenge resulted in isolation of L. monocytogenes from 11% of both left and right knee cultures. These results suggest that L. monocytogenes Scott A colonization of turkey knee synovial tissue can initiate in day-of-age poults and that L. monocytogenes Scott A can be invasive through air sac infection.     

Vaccination of pregnant ewes against infection with Salmonella Brandenburg

AIMS: To develop a challenge model for Salmonella Brandenburg infection in pregnant ewes. To compare efficacies of a live attenuated Salmonella Typhimurium mutant, a subunit preparation from a virulent S. Brandenburg isolate, and a commercial multivalent inactivated vaccine in their ability to prevent experimental S. Brandenburg infection. To assess the efficacy of the live attenuated S. Typhimurium mutant against natural S. Brandenburg infection in lambs.

METHODS: Two-year-old ewes were immunised with either a live attenuated vaccine (eye-drop; n=20), a subunit vaccine (n=20) or an inactivated bacterin vaccine (n=20), both administered subcutaneously, or served as unvaccinated controls (n=21). Four weeks later, the sensitising regime was repeated as a booster vaccination, and the ewes were challenged 6 weeks later with a virulent S. Brandenburg isolate, approximately 6 weeks prior to lambing. The presence of clinical signs, abortion or death was noted following challenge. The presence and number of Salmonella spp in faecal samples taken throughout the trial, and in organs collected post mortem, were determined using an enrichment selection procedure, and confirmed by serology and pulsed-field gel electrophoresis (PFGE). Half of the surviving lambs were vaccinated with the live attenuated vaccine and all (n=39) were exposed to natural infection from contaminated pasture.

RESULTS: There was no significant protection against mortality and abortion following vaccination with the live attenuated, subunit and inactivated vaccines following experimental challenge with S. Brandenburg. There was a significant but transient decrease in the number of ewes shedding S. Brandenburg (live attenuated, p=0.05; subunit, p=0.05; inactivated, p=0.01), and in the quantity of these bacteria in the sheep from the vaccinated groups (p<0.05) compared with controls, 6 weeks after challenge. Lambs from the challenged ewes did not shed Salmonella spp after being vaccinated with the live attenuated vaccine, in contrast to some of the controls, when grazed on pasture contaminated with S. Brandenburg.

CONCLUSIONS: The use of live attenuated, subunit and inactivated vaccines did not significantly protect sheep against lethal experimental challenge with S. Brandenburg.     

Vaccination of pregnant ewes against infection with Salmonella Brandenburg

AIMS: To develop a challenge model for Salmonella Brandenburg infection in pregnant ewes. To compare efficacies of a live attenuated Salmonella Typhimurium mutant, a subunit preparation from a virulent S. Brandenburg isolate, and a commercial multivalent inactivated vaccine in their ability to prevent experimental S. Brandenburg infection. To assess the efficacy of the live attenuated S. Typhimurium mutant against natural S. Brandenburg infection in lambs. METHODS: Two-year-old ewes were immunised with either a live attenuated vaccine (eye-drop; n=20), a subunit vaccine (n=20) or an inactivated bacterin vaccine (n=20), both administered subcutaneously, or served as unvaccinated controls (n=21). Four weeks later, the sensitising regime was repeated as a booster vaccination, and the ewes were challenged 6 weeks later with a virulent S. Brandenburg isolate, approximately 6 weeks prior to lambing. The presence of clinical signs, abortion or death was noted following challenge. The presence and number of Salmonella spp in faecal samples taken throughout the trial, and in organs collected post mortem, were determined using an enrichment selection procedure, and confirmed by serology and pulsed-field gel electrophoresis (PFGE). Half of the surviving lambs were vaccinated with the live attenuated vaccine and all (n=39) were exposed to natural infection from contaminated pasture. RESULTS: There was no significant protection against mortality and abortion following vaccination with the live attenuated, subunit and inactivated vaccines following experimental challenge with S. Brandenburg. There was a significant but transient decrease in the number of ewes shedding S. Brandenburg (live attenuated, p=0.05; subunit, p=0.05; inactivated, p=0.01), and in the quantity of these bacteria in the sheep from the vaccinated groups (p<0.05) compared with controls, 6 weeks after challenge. Lambs from the challenged ewes did not shed Salmonella spp after being vaccinated with the live attenuated vaccine, in contrast to some of the controls, when grazed on pasture contaminated with S. Brandenburg. CONCLUSIONS: The use of live attenuated, subunit and inactivated vaccines did not significantly protect sheep against lethal experimental challenge with S. Brandenburg.     

Light and electron microscopic study of the livers of pregnant mice infected with Listeria monocytogenes.

Listeria monocytogenes cells were observed in the hepatic cell cytoplasm or in the phagosome at 24 and 48 hours but not at 72 hours after inoculation in pregnant mice. The presence of bacteria initially in a membrane-bound vesicle indicates that the bacteria enter the hepatic cells by endocytosis, resulting in eventual destruction of hepatic cells. Characteristic lesions of the liver at 24 and 48 hours after inoculation consist of multiple focal areas of necrosis. The initial neutrophilic reaction seems to give way to a mononuclear reaction (listeriomas) at 72 hours after inoculation. Dilation of rough endoplasmic reticulum and release of many of the bound ribosomes with a relative increase in the number of free ribosomes was observed. Hepatic lesions were not observed in control (nonpregnant) mice.     

Clinical and histopathological aspects of naturally occurring mastitis caused by Listeria monocytogenes in cattle and ewes

Listeria monocytogenes was isolated from the milk of two cows and two sheep with mastitis in one quarter and one udder half. The animals were observed over a period of 2-12 months. Clinical examination of the udder, bacteriological examinations and determination of somatic cell counts of milk samples were performed monthly. All four cases suffered from a subclinical mastitis characterized by an elevated somatic cell count (0.8-10.1 x 10(6) cells/ml), a persistent shedding of Listeria and by a normal appearance of the milk. The animals did not show any systemic reaction, but all animals developed an atrophy of the infected mammary gland. Histological examinations revealed a chronic interstitial mastitis with diffuse infiltration of lymphocytes, plasma cells and macrophages. All internal organs showed no abnormalities, no Listeria could be isolated. Listeria could however be isolated from the affected mammary parenchyma and from the mammary lymph node. The results of the bacteriological examination could be confirmed by means of PCR. Using PFGE, all the isolates from the same animal were identical. Immunohistochemical examination of the ovine mammary glands achieved a very strong immunoreactivity for CD5 cells. The mode of infection and the reaction of the immune system's defense of the ovine udders are discussed.     

Prolonged persistence of Listeria monocytogenes after intragastric infection in corticosteroid-treated mice

In an attempt to obtain a model more closely resembling natural listeriosis, we studied the course of infection in mice inoculated by the intragastric route with Listeria monocytogenes. Corticosteroid-treated, and untreated mice both developed subclinical infection without mortality, but faecal shedding and peristence of bacteria in the liver and spleen of corticosteroid-treated mice were significantly more protracted than in untreated mice. Untreated mice cleared the bacteria from their livers and spleens by day 5 postinfection (PI), whereas treated mice did not clear the organisms until 8–9 days PI. In untreated mice faecal shedding lasted 5 days PI, whereas in treated mice the organisms were recovered at significantly higher levels until day 9 PI. The only intestinal lesions observed were mild pyogranulomatous changes in the dome area of some Peyer's patches in treated mice.     

Experimental infection of pregnant goats with swine fever virus

Thirteen pregnant goats were inoculated intravenously with the ALD strain of virulent swine fever (SF) virus on Days 64-84 of gestation. Dams showed transient and mild viremia, and produced high serum neutralizing (SN) antibody after inoculation. Six inoculated dams were reared until parturition occurred and bore six apparently normal, one apparently normal but dead, one mummified and three edematous kids. Neutralizing antibody was demonstrated in the pre-colostral sera obtained from all normal kids, but no SF virus was isolated from any of them. The other seven dams were killed on post-inoculation days (PID) 5-61, and fetuses, placenta and amnion were tested for the virus and SN antibody. All fetuses of five dams examined within PID 40 were positive for SF virus, but negative for SN antibody. SF virus was also isolated from one of three fetuses examined on PID 61. Conversely, the other two fetuses examined on PID 61 were negative for SF virus, but positive for SN antibody. Furthermore, SF virus was isolated from the placenta and amnion of all the dams.