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.     

Fish oil-induced yellow fat disease in rats. I. Histological changes

Yellow fat disease was induced in young rats given a vitamin E-deficient diet supplemented with 15% fish oil. The changes in adipose tissue of this oil-induced disorder were different from those of natural yellow fat disease in horse, pig and mink. In the natural disease all fat depots had the early stage of yellow fat disease with interstitial lipofuscin-laden macrophages exclusively. In the rat, however, this change was seen only in the subcutaneous fat depot. Moreover, affected adipose tissue of animals with natural disease had extensive fibrosis, but in the rat fibrosis was always absent. Rats with fish oil-induced yellow fat disease had degenerative changes in various fat depots that occurred at various times but in the horse, pig and mink fat depots were affected simultaneously. Lipofuscin accumulated in the reticuloendothelial system in rats. Accumulation in spleen and liver was dependent on vitamin E deficiency, but only the accumulation in the Kupffer cells was correlated with yellow fat disease. Lipofuscin accumulation in the mesenteric lymph node did not depend on vitamin E deficiency.     

Reduced in vivo nonspecific resistance to Listeria monocytogenes infection during avian retrovirus-induced immunosuppression

Ten-day-old chickens infected with an avian osteopetrosis virus [MAV-2(O)] were more susceptible to challenge with Listeria monocytogenes than virus-free chickens, as demonstrated by reduced bacterial clearance from their spleens. Reduced clearance of L. monocytogenes was observed throughout a 26-day period after MAV-2(O) infection.     

Influence of liver fat on experimental Escherichia coli mastitis in periparturient cows

Eleven cows with a wide range of liver fat (5.7 to 51.4 per cent) at seven days post partum were experimentally infected in a single quarter with a capsular Escherichia coli at 10 days post partum. The results suggested that a fatty liver in itself does not influence the severity of mastitis. All animals had clinical mastitis 10 hours after infection but no animals became severely ill and no treatment was given. Four out of five animals in the group with less than 20.2 per cent liver fat had bacteria in their milk at 10 hours after infection but these bacteria were eliminated by 12 hours. The six animals in the group with more than 28.3 per cent fat in their liver retained viable bacteria in the udder for much longer; with two animals bacteria were shed and abnormal milk was secreted for up to four months despite antibiotic therapy.     

Fish oil-induced yellow fat disease in rats. II. Enzyme histochemistry of adipose tissue

Adipose tissue in various stages of fish oil-induced yellow fat disease in the rat had the same acid phosphatase and 5-nucleotidase activity pattern as similar stages of the disorder in mink and pig. A weak acid phosphatase and 5-nucleotidase activity was seen in interstitial lipofuscin-laden macrophages in "stage M" yellow fat disease without fat cell degeneration. Activity of these macrophagic enzymes increased when there was fat cell degeneration ("stage S" and "stage E" yellow fat disease). This different phosphatase activity in the same cell type may result from phagocytosis of substrates with variable digestibility. Macrophages directly surrounding affected fat cells in steatitis areas ("stage S" and "stage E") had strong acid phosphatase and 5-nucleotidase activity. As in the pig, increased 5-nucleotidase activity was found in affected fat cells, which probably indicates plasma membrane damage. Increased nonspecific esterase activity occurred around affected fat cells. Only a small part of this esterase activity originated from inflammatory cells. This indicates that an increase of esterase activity in degenerating adipose tissue may be an endogeneous process in this tissue.     

产后奶牛子宫内微生物菌群动态变化及其与子宫黏液性状关系的研究

本文旨在研究奶牛产后子宫内优势菌群随时间的动态变化及细菌种类与子宫黏液性状之间的关系,以便于临床的快速诊断及治疗。选取分娩日期临近、产后子宫黏液性状不同的荷斯坦奶牛42头,于分娩后10、20、30和40d采集子宫黏液,体外培养法进行细菌分离、鉴定。结果显示,产后奶牛子宫内分离率最高的细菌分别为大肠杆菌(85.7%)、变形杆菌(64.3%)和葡萄球菌(61.9%);除大肠杆菌在整个试验期分离率均较高外,产后10、20、30和40d优势细菌分别为葡萄球菌、产气荚膜梭菌、沙门氏菌和葡萄球菌。白色或灰白色脓性黏液,黄色或土黄色黏液中细菌平均分离株数(5.6±0.4,5.5±1.4)高于清亮透明黏液(3.5±1.6),其中沙门氏菌、变形杆菌、链球菌和化脓隐秘杆菌产后30和40d的分离株数高于清亮黏液,说明其可能与白色或黄色脓性黏液的出现相关;产后40d黄色或土黄色化脓性黏液可能与产气荚膜梭菌的出现相关;而粪肠球菌和嗜酸乳杆菌产后20、30、40d在脓性黏液中分离数高于清亮黏液,说明其作为有益菌可能在分泌不正常黏液的子宫内具有抵抗病原菌的作用。结果表明,产后奶牛子宫内优势菌群呈动态变化,子宫黏液性状与特异病原菌的出现有关。     

A Disease Resembling Paratuberculosis (Johne’S Disease) in Roe Deer (Capreolus Capreolus L.) an Aetiological and Patho-Anatomical Study

A disease in wild living roe deer (Capreolus capreolus L.) caused by acid-fast bacteria is described.The morphological and cultural properties of these bacteria agree closely with corresponding properties of Mycobact. johnei.Enlarged lymph nodes, especially mesenteric lymph nodes with greyish-yellow necroses, were the most prominent macroscopic lesions. No intestinal lesions were present.The histopathological picture of the lymph nodes resembled mostly lesions which can occur in paratuberculous sheep and goats. The epithelioid cells contained masses of acid-fast bacteria. Such rods were also demonstrated in the intestinal villi.The acid-fast bacteria could be isolated from the mesenteric lymph nodes, spleens, bone marrow, mammary gland and also from the lymph nodes of other organs.The organism did not produce tuberculosis in guinea pigs. Intravenous injections into hens and rabbits resulted in miliary nodules resembling those of tuberculosis in the livers and spleens of the hens, and in joint and tendon sheath lesions in the rabbits. Microscopically the lesions mostly resembled those in paratuberculosis.One of the strains which was inoculated intravenously into two calves caused no lesions. The results of the allergy tests and serological blood tests in one animal indicate that infection with Mycobact. johnei cannot be excluded.A goat which was inoculated in the same manner and with the same strain as the calves died after six weeks. Miliary epithelioid cell granulomas in the liver, spleen, kidneys, bone marrow of long bones and in the submucosa of the ileum, as well as patchy infiltration of epithelioid cells in, among others, the mesenteric lymph nodes were observed on microscopic examinations.By intravenous infection the disease could be reproduced in a roe deer (fawn) (Gapreolus capreolus L.). The animal died nine weeks after the infection, and during the last two days before death it had a profuse diarrhoea. Masses of short acid-fast bacteria in clumps were present in the faeces. Nor in the experimental animal did macroscopic intestinal lesions occur. Enlarged villi infiltrated with epithelioid cells containing acid-fast bacteria were demonstrated by histological examination.The acid-fast bacteria could be recovered, but only on the Taylor/ Finlayson medium, from all experimental animals except the guinea pigs. Concerning the experimental calves, acid-fast bacteria were recovered from only one of them and then nearly three years after the infection.The acid-fast bacteria did not reduce nitrate. They showed positive neutral red reactions and were sensitive to isoniazid in a concentration of 2.5 µg per ml medium after three weeks’ incubation.The possibility that the isolated acid-fast bacteria and the lesions caused by them might be avian tubercle bacilli and avian tuberculosis has been discussed by the author. He does not, however, find any relevant reason for such an assumption. The author considers that the bacteria and the lesions exhibit the greatest similarity to Mycobact. johnei and paratuberculosis (Johne’s disease).On account of the organism’s pathogenicity for hens and rabbits, and necroses in the course of the disease, the author suggests that these bacteria possibly constitute a variety of the classic bovine Mycobact. johnei, different from the pigmented and the Icelandic varieties.     

The early pathogenesis in chicken inoculated with non-pathogenic serotype 2 Marek's disease virus.

A newly cloned serotype 2 Marek's disease virus (MDV), strain ML-6, was inoculated via the nasal cavity in specific-pathogen-free chicks to examine early virus replication and the expression of Marek's disease (MD)-related antigens. Following inoculation, viral intracellular antigens (VIAs) were detected in lymphoid organs (bursas and spleens) between 5 and 14 days post inoculation (PI), in feather follicles between 14 and 30 days PI, and in lungs at 3 days PI by the immunohistopathological staining of avidin-biotin-peroxidase complex method. But, very few VIAs were expressed in the thymuses between 5 and 14 days PI. However, MD tumor-associated surface antigens were not detected in any organs. Viruses were isolated from separated spleen cells at 14 and 30 days PI. Fluorescent antibodies of convalescent sera were also detected after 10 days PI. As most of the VIAs were detectable in B-cells in bursas and spleens. B-cells were considered to be the main first target cells for the serotype 2 MDV infection.     

The role of nitric oxide in Listeria encephalitis of ruminants and in rats intracisternally infected with Listeria monocytogenes

Listeria monocytogenes is a Gram-positive facultative intracellular bacteria which infects a wide range of hosts. In ruminants, infection with L. monocytogenes frequently causes encephalitis, which is usually fatal in sheep and goat, while cattle often recover with antibiotic therapy. Since the role of NO in the control of Listeria is controversial, we have studied the expression of iNOS in the brains of cattle, sheep and goats which had succumbed to listeria encephalitis. iNOS was demonstrated in decreasing intensity in the M phi of microabscesses from cattle, sheep and goat. iNOS expression was accompanied by NT in the microabscesses of cattle, but was only present to a low degree in sheep and was absent in goats. This is indirect evidence for differences in the ability to produce NO in the three species. Presence of iNOS and NT were inversely correlated with the numbers of bacteria. While microabscesses of goats contained high amounts of L. monocytogenes they occurred only rarely in cattle. To corroborate our hypothesis that NO is involved in the control of listeria encephalitis a new animal model was developed. Eleven day old infant rats were infected intracisternally with a low dose of L. monocytogenes. This resulted in a transient meningoencephalitis with moderate clinical signs and low mortality. Listeria proliferated strongly in the inflammatory lesions during the first days of infection, reached a peak at day 4 and were eliminated until day 7. The presence of bacteria was closely accompanied by high numbers of iNOS-expressing M phi and the formation of NT. Administration of the iNOS inhibitor L-NIL or the radical scavenger PBN resulted in rapid death of the treated animals. However, the increase in bacterial numbers was one order of magnitude higher for animals treated with PBN compared with L-NIL administration. This shows that NO plays an important role in the control of a brain infection with Listeria, but suggests that reactive oxidants other than NO are also involved. In conclusion, our findings point to a possible involvement of the differences in the ability to express iNOS and subsequent NO production in the different clinical outcome of listeria encephalitis in cattle and small ruminants.     

单增李斯特菌srtA基因缺失株的构建及srtA基因对其毒力的影响研究

为了研究srtA 基因对单增李斯特菌LM90SB2毒力的影响,本研究利用同源重组技术构建了LM90SB2 srtA 基因缺失株LM90SB2-ΔsrtA,比较分析了亲本株LM90SB2与缺失株LM90SB2-ΔsrtA对MBMEC、HBMEC、RAW264.7、SIEC 4种细胞系的粘附、侵袭、胞内增值能力及LM90SB2和LM90SB2-ΔsrtA 感染小鼠后,小鼠的LD50及肝脏、脾脏、脑载菌量变化差异。结果显示,本研究成功构建了srtA 基因缺失株;与亲本株LM90SB2比较,缺失株LM90SB2-ΔsrtA 对RAW264.7和SIEC的粘附率、侵袭率及胞内细菌数量均下降,且差异具有显著统计学意义(p <0.05);小鼠 LD50降低了21.38倍,肝脏、脾脏载菌量降低,差异具有显著统计学意义(p <0.05)。本研究结果表明,srtA 基因对单增李斯特菌LM90SB2的毒力具有关键作用,参与粘附、侵袭BMEC,该研究结果为进一步阐明单增李斯特菌毒力因子的致病机制提供理论依据。     

RNaseⅢ RncS对单核细胞增生李斯特菌毒力的调控作用研究

核糖核酸酶RnaseⅢ是一种调控nc RNA水平的重要酶系。为了解核糖核酸酶RnaseⅢRncS在单核细胞增生李斯特菌(LM)毒力中的调控作用,本研究在构建LM-ΔrncS基因缺失突变株的基础上,通过动物感染试验检测强毒株LM-SB5与缺失株LM-Δrnc S对昆明系小鼠的LD50、存活能力、脏器载菌量及病理组织学产生的影响;利用细胞侵染试验检测强毒株与缺失株对小鼠巨噬细胞RAW264.7的粘附率、侵袭率及其在胞内生存繁殖能力的影响,分析RnaseⅢRncS对LM毒力的影响。结果显示,LM-SB5强毒株和LM-Δrnc S缺失株对昆明系小鼠的LD50分别为105.60 CFU、106.90 CFU;与LM-SB5强毒株相比,LM-Δrnc S的LD50升高了1.30个对数数量级,小鼠的存活时间明显延长,表明毒力显著降低;第3~5 d肝脏、脾脏载菌量显著减少(P<0.05),其中第4 d差异极显著(P<0.01);LM-Δrnc S缺失株对肝脏、脾脏、肾脏的病理损伤降低;LM-Δrnc S缺失株对RAW264.7细胞的粘附率和侵袭率均显著低于LM-SB5强毒株(P<0.01),在2~6 h之间,LM-Δrnc S缺失株在细胞内的细菌量显著低于LM-SB5强毒株(P<0.05),证实LM-Δrnc S在细胞内的生存增殖能力显著降低,提示rnc S基因对LM毒力发挥有一定的调控作用。本研究为进一步揭示RnaseⅢ在LM毒力中的分子调控机制提供了科学依据。     

The effects of parenteral iron dextran and/or desferrioxamine on the development of experimental pseudotuberculosis in the domestic chicken (Gallus domesticus)

Abstract The development of disease following oral challenge with Yersinia pseudotuberculosis (serotype 11) was compared in four groups of five birds treated with a parenteral dose of 10 mg iron dextran (Imferon), 10 mg of iron dextran plus 10 mg of the chelating agent desferrioxamine (Desferal), 10 mg of desferrioxamine or 10 mg of dextran 2 days before the experiment. Four groups of two birds received the above treatment regimens but no bacterial challenge. In iron dextran treated birds, oral challenge resulted in faecal shedding for the 10 day duration of the experiment, whereas in those birds which received dextran or desferrioxamine alone, the duration of faecal shedding was significantly less. Serological titres to the lipopolysaccharide antigen of the challenge bacteria were also lower in the groups not pretreated with iron dextran. The birds pretreated with iron dextran had diarrhoea and were clinically unwell 2 days following the initial oral challenge. Birds not given iron dextran showed no clinical signs of disease. Histological examination of five selected areas in the liver, spleen and intestine of each bird indicated that birds in the groups treated with iron dextran prior to bacterial challenge had significantly more intestinal lesions than birds in the groups not treated with iron. In contrast, there were significantly more lesions in the spleens of birds not pretreated with iron dextran. There was no evidence of stainable iron in the livers of birds challenged with Y pseudotuberculosis 10 days after an injection of 10 mg of iron dextran. This is in contrast to birds given iron dextran and no bacteria. It was concluded that pretreatment of birds with iron dextran resulted in more severe clinical disease, prolonged faecal shedding with associated intestinal lesions and higher serological titres to bacterial antigen. The number of lesions in the spleen and liver was not necessarily correlated with the severity of clinical disease, and in all infected birds the hepatic iron levels were significantly lower than in the non-infected control birds 10 days after oral challenge. It seems probable that the chicken has a high requirement for iron during infection with Y pseudotuberculosis and mobilises stored and exogenously supplied iron for tissue repair and immunological function.     

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.     

Avirulence of viable but non-culturable Listeria monocytogenes cells demonstrated by in vitro and in vivo models

The virulence of Viable But Non-Culturable (VBNC) cells of 4 strains of Listeria monocytogenes was investigated in both a human adenocarcinoma cell line (HT-29) and a mouse model. LO 28, ATCC 19115 and CNL 895807 strains of Listeria monocytogenes became VBNC when incubated in microcosm water at 20 degrees C and Scott A strain at 4 degrees C. No culturable bacteria were detected in the VBNC state, although 104 active cells/mL were found by the Direct Viable Count (DVC) and CTC-DAPI double staining methods. A comparison of virulence in both human adenocarcinoma cell line HT-29 and the mouse model showed that culturable controls were more virulent than VBNC cells, which appeared to be avirulent regardless of the virulence methods applied. Pathogenicity was tested in each model and was lost concomitantly with culturability, whereas some cells were still metabolically active (determined by CTC and DVC). Moreover, amplification of a 388 bp fragment with Immunocapture-PCR revealed the presence of Listeria monocytogenes DNA in all mixed spleen samples after intravenous injection of VBNC cells. These results demonstrate that VBNC cells were present in the mouse spleens. The results of the study suggest that Listeria monocytogenes strains might remain in the aquatic environment for prolonged periods in the VBNC state but these cells were not pathogenic in the conditions tested. These findings demonstrate the value of VBNC studies and show the need to investigate the role of VBNC cells in environmental transmission of Listeria monocytogenes. Further studies are needed in order to investigate the virulence of VBNC cells of Listeria monocytogenes after recovery of a culturable state.     

Differential effect of T-2 toxin on murine host resistance to three facultative intracellular bacterial pathogens: Listeria monocytogenes, Salmonella typhimurium, and Mycobacterium bovis

The effect of T-2 toxin, a radiomimetic immunosuppressive agent, on resistance to the facultative intracellular bacterial pathogens Listeria monocytogenes (strain EGD), Mycobacterium bovis (BCG Copenhagen 1331), and Salmonella typhimurium was determined. Female Swiss ICR mice were given a single dose of T-2 toxin (4 mg/kg of body weight) by gastric gavage. On the seventh day after toxin administration, the mice were infected by intraperitoneal inoculation with L monocytogenes, S typhimurium, or M bovis. Mice given the toxin also were exposed to respirable droplet nuclei containing L monocytogenes or M bovis. The effect of the toxin on the course of infection was monitored by observing mortality or by enumeration of bacteria in the spleen or lungs of infected mice. The toxin increased resistance to infection with L monocytogenes initiated by intraperitoneal inoculation, but reduced resistance to M bovis infection initiated by intraperitoneal inoculation. The toxin had no appreciable effect on the course of salmonellosis or on resistance to infection initiated by inhalation of L monocytogenes or M bovis aerosols. Therefore, it was concluded that T-2 toxin does not necessarily reduce resistance to infection in mice. The toxin's effect on the course of in vivo bacterial infections depends on the nature of the infective agent and the route of inoculation.     

The effects of parenteral iron dextran and/or desferrioxamine on the development of experimental pseudotuberculosis in the domestic chicken (Gallus domesticus)

The development of disease following oral challenge with Yersinia pseudotuberculosis (serotype 11) was compared in four groups of five birds treated with a parenteral dose of 10 mg iron dextran (Imferon), 10 mg of iron dextran plus 10 mg of the chelating agent desferrioxamine (Desferal), 10 mg of desferrioxamine or 10 mg of dextran 2 days before the experiment. Four groups of two birds received the above treatment regimens but no bacterial challenge. In iron dextran treated birds, oral challenge resulted in faecal shedding for the 10 day duration of the experiment, whereas in those birds which received dextran or desferrioxamine alone, the duration of faecal shedding was significantly less. Serological titres to the lipopolysaccharide antigen of the challenge bacteria were also lower in the groups not pretreated with iron dextran. The birds pretreated with iron dextran had diarrhoea and were clinically unwell 2 days following the initial oral challenge. Birds not given iron dextran showed no clinical signs of disease. Histological examination of five selected areas in the liver, spleen and intestine of each bird indicated that birds in the groups treated with iron dextran prior to bacterial challenge had significantly more intestinal lesions than birds in the groups not treated with iron. In contrast, there were significantly more lesions in the spleens of birds not pretreated with iron dextran. There was no evidence of stainable iron in the livers of birds challenged with Y pseudotuberculosis 10 days after an injection of 10 mg of iron dextran. This is in contrast to birds given iron dextran and no bacteria. It was concluded that pretreatment of birds with iron dextran resulted in more severe clinical disease, prolonged faecal shedding with associated intestinal lesions and higher serological titres to bacterial antigen. The number of lesions in the spleen and liver was not necessarily correlated with the severity of clinical disease, and in all infected birds the hepatic iron levels were significantly lower than in the non-infected control birds 10 days after oral challenge. It seems probable that the chicken has a high requirement for iron during infection with Y pseudotuberculosis and mobilises stored and exogenously supplied iron for tissue repair and immunological function.     

Experimental infection of pregnant ewes with Listeria monocytogenes

Eight 18-month-old ewes were infected orally with Listeria monocytogenes between 77 and 91 days of pregnancy. Only one ewe aborted, 10 days after the first infecting dose, at 94 days of gestation; L monocytogenes was isolated from several sites in both its aborted fetuses. Two days after the first infecting dose all the ewes exhibited mild illness and pyrexia lasting for two to three days but the ewe which aborted was seriously ill until nine to 10 days after aborting. Agglutination tests carried out on 2-mercaptoethanol reduced sera revealed a strong immunological response in all the infected ewes but in the ewe which aborted this response was delayed. Four uninfected ewes which were kept as controls remained healthy throughout the experiment and showed no evidence of 2-mercaptoethanol resistant antibodies to L monocytogenes. Growth retardation lines, occurring at the time of and after experimental infection, were found in the bones of 14 of 17 newborn lambs in both the infected and control groups; in the aborted lambs these lines occurred before the infection.     

Lewis rats are not susceptible to oral infection with Mycobacterium avium subsp. paratuberculosis

Pathogenesis studies of Mycobacterium avium subsp. paratuberculosis infection in ruminants are hampered by the long incubation time of the disease. A laboratory animal model with a shorter incubation time would facilitate research in this field. Although small rodents are usually considered to be resistant to M.a. paratuberculosis infection, several susceptible murine strains have been found. To our knowledge, there are no detailed reports with regard to susceptibility in rats. The Lewis rat is a valuable model for inflammatory bowel disease studies as well as autoimmune diseases involving mycobacteria as inducing agents. In this study Lewis rats were used to investigate their potential as a small laboratory animal model for paratuberculosis.In total 28 female Lewis rats were orally inoculated with M.a. paratuberculosis. The rats were first inoculated at 3 weeks of age, and 12 more inoculations followed in increasing intervals during the 3 months to follow. Eight control rats received a sham inoculation. Over 9 months, two rats from each group were sacrificed at regular intervals and immunological and histopathological examinations were performed on the gastrointestinal tract, the liver and the spleen.None of the rats developed lesions which were indicative of mycobacterial infection as determined by histology with HE and Ziehl-Neelsen staining. The bacteria could not be recultured from samples taken from the gut, the liver or the spleen. The immunological tests however, showed that bacteria had entered via the intestinal tract. From this study it appears that Lewis rats are resistant to oral inoculation with M. a. paratuberculosis, and not suitable as a model to study the immunopathogenesis of paratuberculosis as it occurs in ruminants.     

Clinical and serum antibody responses to lambs to infection by Listeria monocytogenes.

Oral dosing of lambs with 1 x 10(10) colony forming units of Listeria monocytogenes daily for three days produced no clinical signs but protected the animals against bacteraemia following subsequent homologous subcutaneous challenge. Following the subcutaneous injections, comparison with controls revealed significantly lower rectal temperatures and a significant difference in positive blood cultures. In both groups signs of systemic illness were unremarkable. However, two and 10 days after the subcutaneous challenges neurological signs developed in two lambs. L monocytogenes was isolated from the brain of one lamb and histopathological lesions of listeric encephalomyelitis were demonstrated in both. After oral infection antibodies to L monocytogenes whole cell antigen were detectable in serum agglutination tests and by ELISA. Serological responses to flagellin were examined by ELISA and to listeriolysin O by immunoblotting. The responses of the animals to flagellin were weak and inconsistent, but antibodies to listeriolysin O were detectable after both oral and subcutaneous challenge. The subclass of antibody involved in this response was shown to be predominantly IgG1.     

Humoral and Delayed-type Hypersensitive Responses againstListeria monocytogenes Phosphatidylinositol-specific Phospholipase C in Experimentally Infected Buffaloes

The kinetics of antibody production against phosphatidylinositol-specific phospholipase C (PI-PLC) and the isolation pattern of Listeria monocytogenes from bacteriological samples were studied following oral infection of buffalo calves with 3 x 10(9) cells each of pathogenic L. monocytogenes. Antibodies to PI-PLC appeared by 4-8 days post infection (PI), with a peak between days 7 and 16 PI, when tested by indirect plate-ELISA. Subsequently, antibody titres in all the animals declined and became undetectable on days 26-35 PI onwards until the study concluded on day 211 PI. Dot-ELISA could detect the antibodies to PI-PLC 1-2 days earlier and at higher titres as compared to plate-ELISA. L. monocytogenes could be recovered from faeces, nasal swabs and haemocultures from days 2 to 33, days 2 to 21 and days 11 to 17 PI, respectively. Antibodies to PI-PLC were detected during the course of active infection but their titres declined sharply once animals became culturally negative. Sonicated antigen elicited the highest delayed-type hypersensitivity response, followed by PI-PLC and listeriolysin O.