T cell-mediated immunity to Cowdria ruminantium in mice: the protective role of Lyt-2+ T cells

The inability of athymic nude mice to make a drug-aided recovery from infection with either the Kümm or the Welgevonden stocks of Cowdria ruminantium and their inability to mount an immune response, suggest that immunity in heartwater is cell-mediated. The adoptive transfer of immunity with the spleen cells of mice immune to the Welgevonden stock is supportive evidence. Immune spleen cells depleted of Lyt-2+ T cells are unable to confer resistance to challenge to recipient mice, whereas the depletion of L3T4+ T cells had no effect on the protection conferred by immune spleen cells. This is conclusive evidence that immunity in heartwater is largely cell-mediated. Immune serum, C. ruminantium and complement incubated in the presence of mouse peritoneal macrophages, inhibits the infectivity of the heartwater agent, but not in the absence of macrophages. The decreased resistance to challenge of immune mice treated with gloxazone adds further support to the concept that in heartwater persistence of C. ruminantium in the host is associated with immunity.     

An attempt to improve the immunization of sheep against heart-water by using different combinations of 3 stocks of Cowdria ruminantium

Neither sheep immune to the Ball 3, the Kwanyanga or the Mara stocks of Cowdria ruminantium nor those immunized with combinations of these 3 stocks were protected against challenge with the Mali stock. Against challenge with the Welgevonden stock, however, immunization with each of the 3 combinations and with single stocks effected a protective immunity.     

Serum antibody response to Mycoplasma hyopneumoniae measured by enzyme-linked immunosorbent assay after experimental and natural infection of pigs

Studies were conducted under experimental and field conditions to determine the effect of infection with M. hyopneumoniae on the immune response in serum as measured by ELISA. Following intratracheal challenge or contact exposure, serologically negative pigs derived from mycoplasma-free piggeries developed an immune response within 10 days. This response continued to rise for a further 50 days. In a field study in a commercial piggery, no animals (0/44) were observed to have M. hyopneumoniae antibodies at day 86 of life. However between day 86 and day 144, 97.7% (42/43) animals sero-converted. These results are discussed in terms of infection spread, particularly in the grower/finisher shed.     

Antigenic differences between stocks of Cowdria ruminantium

Stocks of Cowdria ruminantium from Senegal, Zambia and South Africa were compared in cross immunity tests in goats. The Senegal stock caused fatal heartwater in three of 10 goats immune to the South African reference stock Ball 3, and five others showed significant febrile reactions and recovered spontaneously. Four goats immune to the Senegal stock did not show any reaction on challenge with Ball 3. The stock from Zambia was fully cross-protective with Ball 3 in experiments with three goats, but these three goats, immune to the Zambia stock and to Ball 3, showed severe febrile responses upon further challenge with the Senegal stock. The Senegal stock was highly virulent for Dutch goats and there were exceptionally large numbers of rickettsiae in brain capillaries after death. This stock has been passaged eight times in mice, without causing disease; the presence of the organism in the mice was shown by subinoculating goats. The Senegalese stock of C ruminantium is the first stock outside South Africa against which the reference Ball 3 stock does not fully immunise.     

Immunosuppression of carp (Cyprinus carpio L.) to ichthyophthiriasis using the corticosteroid triamcinolone acetonide

The immune response of juvenile mirror carp (Cyprinus carpio L.) has been investigated in relation to protective immunity and immunosuppression to the disease ichthyophthiriasis. Protective immunity was induced by exposing juvenile carp to approximately 2,000 theronts per fish for 3 hours at a concentration of approximately 80 theronts cm-3 in dechlorinated water pH 7.0-7.2, 20 +/- 2 degrees C in the dark, on three separate occasions of 14 day intervals. Following each exposure, re-infection was prevented by transferring fish to clean aquaria twice daily from day 5-10. Fish were challenged 4 weeks following exposure to the third immunising infection with a potentially lethal dose of approximately 8,000 theronts per fish for 3 hours at a concentration of approximately 320 theronts cm-3. All immunised fish survived although 19% showed initial signs of invasion, the parasites in these instances being confined to periphery of fins. Mortalities of 100% were recorded in non-immunised controls, first deaths occurring 5 days after challenge. Fourteen days following challenge, immune fish received one of the following treatments; intraperitoneal injection of the corticosteroid triamcinolone acetonide at a dose of 200 micrograms g-1 body weight, intraperitoneal injection of 0.85% saline or left untreated; non-immunised fish were subjected to similar procedures. On challenge 6 days later, all fish developed heavy infection with up to 100% mortalities with the exception of immunised fish administered saline or left untreated. Repeat experiments gave comparable results. The results indicate that immunosuppression can be induced by administration of high levels of synthetic corticosteroid.(ABSTRACT TRUNCATED AT 250 WORDS)     

Testing the immunity to orthopoxviruses in the white mouse with vaccinia virus

An infection model was developed, which allows the study of humoral and cellular immune response mechanisms induced by Orthopox viruses in mice. The optimal infection route for neurovirulent vaccinia virus strains was investigated and resulted in well-defined clinical symptoms in non-immunized susceptible mice. Signs of disease were taken as a basis for comparison. Challenge infections by intracutaneous (i.c.) and intraperitoneal (i.p.) application of vaccinia strain Munich 1 (M1) appeared to be most suited to testing immunities of different status in vivo. Mice passively immunized with an anti-vaccinia immune serum survived intraperitoneal challenge infection with 4LD50/mouse. After an intracutaneous challenge infection with 10(4) TCID50/animal, however, they were fully susceptible. Mice immunized with live vaccinia virus showed a solid immunity to both intracutaneous and intraperitoneal challenge.     

Recombinant bovine interleukin-12 stimulates a gut immune response but does not provide resistance to Cryptosporidium parvum infection in neonatal calves

This study was undertaken to determine if administration of recombinant bovine interleukin-12 (rBoIL-12) could stimulate a cellular immune response that protected calves from an oral challenge inoculation with Cryptosporidium parvum oocysts. In a first experiment, rBoIL-12 intraperitoneally administered as a single dose 1 day before challenge inoculation, did not alter the course of infection. The percentage of immune competent cells and levels of cytokine gene expression in the ileo-cecal mucosa and in the draining lymph nodes of treated calves were similar to those of untreated control calves. However, when rBoIL-12 was subcutaneously administered daily from 2 days before infection to 2 days after infection, a consistent increase of T lymphocytes and an higher expression of interferon-gamma (IFN-gamma) was detected. Again, treatment did not alter the course of infection. Similar results were obtained when rBoIL-12 was administered daily for 4 days beginning 2 days after oral inoculation. These data indicate that although rBoIL-12 stimulated a strong immune response in the gut of neonatal calves, the response was not able to provide protection from challenge inoculation with C. parvum oocysts.     

Comparison of the kinetics of the specific cellular immune response to duck hepatitis B virus in infected and immune ducks.

The kinetics of the cell mediated immune response by ducks acutely and chronically infected with, or immune to infection by duck hepatitis B virus (DHBV) was determined. This was measured by an antigen specific blastogenesis assay to duck hepatitis B surface antigen (DHBsAg) and duck hepatitis B core antigen (DHBcAg) using peripheral blood mononuclear cells (PBMC). The three outcomes of acute infection by DHBV were either clearance from both serum and liver, clearance from serum but not liver, or the development of persistent viraemia. Acutely infected ducks that failed to clear the infection also failed to develop a significant cellular immune response to both antigens. Ducks with chronic infection acquired as neonates or as the result of the failure to clear acute infection had an increasing cellular immune response over time. Two groups of immune ducks were examined. These were either ducks that had become immune following infection or that had been vaccinated. Both groups of ducks demonstrated significant cellular responses following challenge with DHBV irrespective of the level of their responses before challenge. However, there was a reduction in the response of their PBMC over a 4-week-period postchallenge. The range of cellular immune responses to DHBV antigens observed in this study has a number of counterparts in hepatitis B infection of humans. Coupled with the defined clinical outcomes that can be established in the duck/DHBV model, further study of the cellular immune response to DHBV is warranted.     

Anticoccidial Vaccination of Broiler Chickens in Various Management Programmes: Relationship between Oocyst Accumulation in Litter and the Development of Protective Immunity

Paracox anticoccidial vaccine was administered to a 7-day-old flock of commercial broiler breeder stock subsequently reared to point-of-lay in the same house. For comparison, three subgroups of another flock of broiler breeders were also vaccinated with Paracox at 7 days of age, reared to 42 days and then transferred to new litter on another farm until point-of-lay. The first subgroup received no further treatment, but the second and third each received a second vaccination with Paracox, either immediately after transfer to the new litter or 42 days after transfer. Using an Eimeria necatrix model, protective immunity was demonstrated by virulent challenge of samples of birds from all groups by the age of 37–40 days (30–33 days after the first vaccination), and was maintained to at least 122–125 days of age, whether the birds remained on the same litter or were transferred to another farm, and whether they received one or two anticoccidial vaccinations. Therefore, there is no disadvantage in transferring birds onto new litter 35 days after a single Paracox vaccination, nor is there any advantage in giving a second vaccination after such a transfer. Vaccinated birds seeded the new litter with oocysts, despite being clinically immune to coccidiosis. A supplementary laboratory experiment showed that birds vaccinated at 8 days of age passed almost no oocysts after a second vaccination at 43 days of age. This indicated that they were not only protected against clinical coccidiosis, but were almost solidly immune to a homologous infection 5 weeks after a single vaccination. Nevertheless, oocysts appeared in the litter of all four groups of commercial breeders throughout the trial, showing that wild-type heterologous infections occurred whether the birds were transferred to new litter or not, but these did not overwhelm the acquired protective immunity and cause clinical coccidiosis.     

Establishment of Ascaris suum in the pig: development of immunity following a single primary infection

Development of immunity after a single primary infection of Ascaris suum in pigs was investigated with regard to the worm population dynamics of a superimposed A. suum infection, host immune response and gross liver pathological changes. Group A was given a primary infection of 60,000 infective A. suum eggs and group B was left uninfected. Four weeks later both groups A and B were inoculated with 1,000 A. suum eggs, and subgroups were slaughtered 7, 14 and 21 days post challenge infection (p.c.i.). An uninfected control group C was slaughtered on day 21 p.c.i. The challenge worm recovery in group A was reduced compared to group B by 12%, 50% and 75% on day 7, 14 and 21 days p.c.i., respectively. In both groups was the expulsion of worms initiated between day 14 and 21 p.c.i. However, in group A the worms were recovered more posteriorly in the small intestine and 21 days p.c.i. the mean worm length was significantly shorter than in group B (p = 0.01). The results above were associated with significantly higher (p < 0.05) antibody response and higher eosinophil counts in group A compared to group B. The present results suggest that the larval growth and survival of a challenge infection are decreased, probably due to higher antibody and eosinophil attack during the migratory phase.     

牛病毒性腹泻病毒对新西兰白兔致病性分析及E2蛋白免疫效果的评价

【目的】 研究牛病毒性腹泻病毒（BVDV）感染对新西兰白兔的致病性以及BVDV E2重组蛋白的免疫效果。【方法】 将实验室培养保存的BVDV病毒纯化并按照Reed-Muench法测定其病毒滴度。在致病性试验中,将10只新西兰白兔随机分为感染组和对照组,每组5只。感染组用1 mL纯化的BVDV病毒攻毒（滴鼻500 μL、耳缘静脉注射500 μL）,对照组用等体积的生理盐水处理,连续3 d,每天1次,每天观察各组兔的临床症状并测量体温;分别于接种病毒后第6、9、12、15、17天通过耳缘静脉采集血液检测血常规;感染病毒第17天采集鼻拭子进行RT-PCR鉴定,采集后剖杀并采集气管、肺脏、脾脏和小肠组织,制备病理切片观察病理变化。在免疫效果评价试验中,将10只新西兰白兔随机分为免疫组和对照组,每组5只,免疫组用E2重组蛋白（1 mg/只）与佐剂混合后经肌内多点注射免疫新西兰白兔,对照组接种等体积生理盐水;共免疫2次,2次免疫间隔为14 d。在一免后0、7、14、21、28 d采集血清,通过间接ELISA方法检测血清中抗重组蛋白特异性抗体水平;在一免后第28天按致病性试验中方法攻毒,在攻毒第17天采集鼻拭子进行RT-PCR鉴定,采集气管、肺脏、脾脏和小肠组织制备病理切片观察病理变化及免疫组织化学检测。【结果】 纯化后BVDV的病毒滴度为4.16×10⁶ TCID₅₀/mL。与对照组相比,感染组部分新西兰白兔6 d内活动减少,采食略微减少,6 d后逐渐恢复正常,在感染第13天出现腹泻症状,从第5天开始体温略微升高,但均在正常范围内波动。与对照组相比,在攻毒第6和9天,感染组白细胞和血小板分别显著和极显著降低（P<0.05;P<0.01）;在攻毒第12、15和17天,感染组白细胞、血小板和淋巴细胞均极显著降低（P<0.01）。鼻拭子RT-PCR检测为阳性,气管、肺脏、脾脏及小肠组织表现出轻度至重度的组织病理学变化。间接ELISA检测结果表明,在一免后7 d时,血清抗体滴度为1：16~1：32;在一免后28 d时,血清抗体滴度为1：256~1：512;免疫攻毒组新西兰白兔鼻拭子经RT-PCR检测为阴性;组织病理学观察显示,免疫攻毒组气管及肺脏表现出轻微的组织病理学变化。免疫组化检测结果显示,免疫组结果均呈阴性,对照组结果均为阳性。【结论】 通过滴鼻及耳缘静脉注射BVDV的方式可以构建新西兰白兔致病模型,BVDV E2亚单位疫苗能够刺激机体产生特异性抗体,起到免疫防御的作用。     

Pre-exposure of cattle to drug-abbreviated Fasciola hepatica infections: the effect upon subsequent challenge infection and the early immune response

In this study we examined whether juvenile liver flukes are capable of stimulating protective immune responses in cattle. Four experimental groups of cattle were studied as follows: group A, a positive control, received a primary infection on day 0 and a secondary infection 28 days later; group B also received two infections but the primary infection was terminated by drug treatment on day 5; group C, received infections on days 0, 5 and 10 which were terminated by drug treatments on days 1, 6 and 11 and then a secondary infection on day 28; group D received an infection only on day 28. Juvenile flukes appear to induce protective responses because: (a) group B animals had significantly lower levels of gamma-GT (P<0.05) than group D; (b) both groups B and C exhibited lower parenchymal phase GLDH levels (P=0.006 and 0.041, respectively); and (c) both groups B and C had lower secondary phase eosinophilia (P=0.002 and 0.02, respectively) than those in group D. Sera taken from groups A-C contained antibodies reacting to a variety of proteins in adult fluke somatic antigen and excretory-secretory preparations, particularly to proteins of 52-60, 68-72 and 82-96 kDa. After secondary challenge the antibody responses of group A to these proteins declined while reactivity to proteins of 28-30 kDa increased. Antibody responses to the 28-30 kDa proteins were not detected in groups B-D until 3 weeks later than those observed in group A. Antibody responses to Fasciola hepatica cathepsin L proteases, which are known to induce protection, were monophasic, of the IgG1 isotype only and were not observed prior to secondary challenge in any of the four groups. In contrast, the response to another protective antigen fraction, a high molecular sized haem protein, was of a mixed IgG1/IgG2 nature and was detected within 14 days of primary infection. However, no significant difference in antibody titres to either protein preparation was observed after the secondary infection when groups B and C were compared to group D.     

Intestinal permeability to macromolecules in piglets infected with transmissible gastroenteritis virus

The permeability of the intestine of specific pathogen free piglets was investigated by measuring the concentration of 125-I in the blood after oral administration of 125-I polyvinylpyrrolidone (125-I PVP, MW=40 000 Da) and the concentration of 131-I in the faeces after intravenous administration of 131-I porcine albumin (131-I PA, MW=68 000 Da). The tests were performed one day before and up to two days after the piglets were infected with the Miller strain of transmissible gastroenteritis (TGE) virus. Biopsies of the jejunum were taken at the end of the experiment and blood samples were taken six-hourly. The piglets became anorexic and had diarrhoea 12 hours after infection; the packed cell volume decreased and the concentrations of urea and total serum proteins increased slightly after infection. However, the marked villous atrophy was not accompanied by an increased permeability of the intestine to PVP or PA.     

Assessment of the immune response to duck plague vaccinations

An experiment was conducted to assess the immune responses of ducks to duck plague (DP) vaccinations employing one commercial and one laboratory-adapted (LA) DP vaccines. Virus neutralisation and leucocyte migration-inhibition tests were conducted at regular intervals before and after vaccinations. Similarly, ducks in vaccinated and control groups were subjected to challenge infection with virulent DP virus.The commercial vaccine yielded a poor immune response and partial protection on challenge whereas satisfactory responses were obtained in ducks receiving two doses of LA vaccine. The humoral as well as cellular factors were stimulated indicating possible involvement of both the immune responses in the protection from duck plague.     

Antibody response and viral shedding profile of Egyptian geese (Alopochen aegyptiacus) infected with low pathogenicity H7N1 and H6N8 avian influenza viruses

Egyptian geese (Alopochen aegypticus), a duck species endemic to sub-Saharan Africa and occasionally implicated in the transmission of avian influenza viruses (AIV) to farmed ostriches, were experimentally infected with low pathogenicity H7N1 and H6N8 viruses to assess viral shedding and immune profiles. Following the first infection with H7N1 virus, high titers of virus were shed from both the tracheae and cloacae for at least 7 days postinfection, and tracheal shedding lasting until day 14. All detectable shedding from both tracheae and cloacae had ceased within 28 days of infection. Antibody titers peaked at day 7 postinfection, but the initial immune response was short-lived. Birds that received a second challenge with the homologous H7N1 virus mounted a more robust response that lasted beyond 66 days postchallenge, and H7N1 virus was detected, albeit at much lower levels, until day 28 post secondary infection (psi) in the cloaca and beyond day 28 psi in the trachea. Birds that received an initial infection with H7N1 virus were also challenged with H6N8 virus, and because a comparable shedding pattern to the H7N1 challenge group was observed, we concluded that the effect of any nonspecific immunity was negligible.     

Effects of chloramphenicol on the development of immune responses to canine distemper virus in Beagle pups

The effect of oral chloramphenicol (CHPC) on the development of immune responses to canine distemper virus (CDV) in Beagle pups was studied. Dogs were treated with CHPC for 14 days at a dose of 50 mg/kg, three times a day. Hematologic changes in CHPC-treated dogs included: polychromasia, anisocytosis, and target cell formation of red blood cells concurrent with vacuolation of lymphocytes and basophilic granule formation in neutrophils. Dogs given this therapy showed normal in vivo and in vitro immune responses after CDV vaccination and survived a virulent CDV challenge, whereas untreated, unvaccinated dogs became ill or died after challenge exposure. The results of this study indicate that CHPC therapy does not interfere with either the prechallenge immune response to attenuated viral antigen or the efficient immune mechanisms invoked during virulent virus challenge.     

Immunisation of cattle against theileriosis in Coast Province, Kenya: laboratory evaluation of a Theileria parva parva stabilate for use in 'infection and treatment' immunisation in the field

Theileria parva parva Marikebuni stock, previously shown to give good protection to immunised cattle in Kilifi District, Coast Province of Kenya, was chosen for large scale immunisation in the district. A large sporozoite stabilate was prepared and evaluated for efficacy and safety in the 'infection and treatment' method, using a long or short acting formulation of oxytetracycline. Susceptible cattle were infected with selected doses of stabilate (10(0), 10(-1), 10(-1.7) and left either as untreated controls, or treated with one of the two oxytetracycline formulations. It was concluded that stabilate dilution at 10(-0.7) or 10(-1) in combination with either formulation of oxytetracycline would effect satisfactory immunisation. The short acting oxytetracycline treatment was judged to be the most efficacious in protecting cattle against homologous challenge. On heterologous challenge it was found that T p parva Marikebuni immune cattle were protected against seven T p parva stocks from Kilifi District and also against four stocks of T p parva from other areas of Kenya. In addition, the Marikebuni stock provided partial protection against challenge by T p lawrencei stocks. Furthermore, cattle immune to T p parva and T p lawrencei were protected against lethal challenge of T p parva Marikebuni stock. Thus, it appears that large scale immunisation of cattle against theileriosis in Kilifi District could be undertaken using the Marikebuni stock. With continued assessment, this stock could provide a master theilerial stock for immunisation against cattle theileriosis in areas free of buffaloes elsewhere in Kenya.     

Resistance to velogenic Newcastle disease virus in leghorn chickens by use of prophylactic lymphokines

A group of 1-day-old commercial leghorn chickens was prophylactically treated with lymphokines obtained from lymphocyte cultures of chickens previously infected with Salmonella enteritidis (S. enteritidis-immune lymphokines [SE-ILK]) with the objective to investigate the effect of SE-ILK on development of Newcastle disease (ND) infection caused by Chimalguacan strain, a Mexican velogenic ND virus (vNDV). Clinical signs, histologic lesions, and hemagglutination-inhibition (HI) serum titers were compared with four other groups, namely, chickens without SE-ILK treatment with virus challenge; with SE-ILK without virus challenge; with nonimmune lymphokine (NILK) treatment and virus challenge; with lymphokine treatment and no virus challenge. SE-ILK was administered intraperitoneally in a dose of 0.5 ml/chicken and was followed 30 min later with the challenge of vNDV in a dose of 10(7.6) 50% embryo lethal dose/ml per bird. Birds were observed during 21 days of postchallenge. Detection of histologic changes and virus isolation procedures were carried out on the third, seventh, 14th, and 21st postinoculation days. HI tests were performed first before treatment and later on the days of histologic sample collection except on the third postinoculation day. Results showed that SE-ILK administration conferred resistance to the chickens because: 1) it significantly diminished the severity of ND infection by inhibiting appearance of clinical signs (P < 0.001), lesions (P < 0.005), and histopathologic changes (P < 0.005); 2) it decreased vNDV isolation rate from the organs (P < 0.001), and 3) it potentialized and even accelerated (P < 0.005) primary immune response by antibodies in the presence of vNDV.