猪伪狂犬病灭活疫苗(HB-J株)对兔与猪免疫攻毒保护平行关系的研究试验

用研制的连续3批猪伪狂犬病灭活疫苗(HB-J株)分别以不同剂量免疫健康兔和断奶仔猪,检测其免疫28 d后的血清抗体,采用血清中和试验法测定血清中和指数,并对免疫28 d后的兔和断奶仔猪分别进行攻毒,统计各组兔和断奶仔猪免疫后攻毒保护率。结果表明,兔免疫28 d后的血清抗体中和指数为794～1258时保护率为60%～80%,血清抗体中和指数≥1479时可获得100%保护;仔猪免疫28 d后的血清抗体中和指数为229～269时保护率为60%～80%,血清抗体中和指数≥331时可获得100%保护。兔与仔猪对猪伪狂犬病灭活疫苗(HB-J株)均产生良好的免疫应答反应,抗体值较高者获得保护率相对较高,兔与猪免疫与攻毒保护呈现平行关系。     

Avian paramyxovirus type 1 infections of racing pigeons: 4 laboratory assessment of vaccination

The immune response and protection from challenge afforded to adult pigeons by four different vaccination schedules were assessed. Intravenous challenge with a field pigeon isolate was done four weeks after the second of two doses of vaccine given four weeks apart. Little difference in protection was seen between two 0.25 ml and two 0.5 ml doses of oil emulsion vaccine, although the latter produced a slightly higher immune response. In both cases one of 10 challenged pigeons became sick and died. One dose of Newcastle disease virus B1 live vaccine followed four weeks later by 0.5 ml oil emulsion vaccine gave a comparable immune response to two 0.25 ml doses of oil emulsion but only six birds survived challenge. Two doses of Newcastle disease virus B1 vaccine gave a poor immune response and little protection from challenge; all 10 birds became sick and eight died. Assessment of the onset of protection following one dose of either 0.5 ml oil emulsion vaccine or Newcastle disease virus B1 indicated some partial protection in the latter group as early as five days after vaccination. Both groups showed protection at 10 days but by 21 days, although protection was sustained in the oil emulsion group, birds receiving live vaccine were fully susceptible. Measurement of the duration of protection in pigeons given two 0.5 ml doses of oil emulsion vaccine indicated that protection had begun to wane by 40 weeks after the first dose.     

猪圆环病毒2型感染对猪瘟疫苗体液免疫应答的影响

采用ELISA方法对单独接种猪瘟疫苗组(CSFV组,n=3)、PCV2感染且出现病毒血症后接种猪瘟疫苗组(PCV2/CSFV组,n=3)及PCV2感染同时接种猪瘟疫苗组(CSFV/PCV2组,n=3)不同时相血清中的猪瘟抗体进行检测;并对PCV2感染对照组(PCV2组)及PCV2/CSFV和CSFV/PCV2组血清中PCV2特异的抗体和核酸分别进行ELISA和PCR检测.结果表明,在接种后52 d CSFV组血清中抗体的阻断值显著高于CSFV/PCV2组(P<0.05);接种后42 d和52 d CSFV组平均抗体效价明显高于PCV2/CSFV和CSFV/PCV2组,其中在52 d CSFV组抗体阳性率这100%(3/3)而PCV2/CSFV和CSFV/PCV2在相应时相抗体阳性率仅为67%(2/3).结果提示PCV2感染可在一定程度上抑制猪瘟疫苗特异性的抗体反应.     

Conjunctivitis caused by a swine Chlamydia trachomatis-like organism in gnotobiotic pigs.

The objective of this study was to determine whether a chlamydial strain recovered from growing and finishing swine with conjunctivitis or keratoconjunctivitis could cause the same infections in gnotobiotic pigs. The strain shares biological characteristics with Chlamydia trachomatis. After propagation in Vero cells and preparation of the inoculum (10(7) inclusion-forming units/ml), chlamydial strain H7 was instilled into the ventral conjunctival sac (0.15 ml/sac) of 12 anesthetized 3-day-old gnotobiotic piglets. Four age-matched gnotobiotic piglets were anesthetized and sham infected with uninfected cell culture lysates. None of the principal piglets developed clinical symptoms of conjunctivitis or keratoconjunctivitis. Principal piglets necropsied 7 days postinfection (DPI) had histologic lesions of mild or moderate conjunctivitis; immunohistochemical evaluation revealed chlamydial antigen in conjunctival epithelium. A majority of principal piglets necropsied at 14-28 DPI had histologic lesions of mild conjunctivitis, but chlamydial antigen was not detected by immunohistochemistry. The results indicated that chlamydial strain H7 can cause mild or occasionally moderate conjunctivitis in gnotobiotic pigs, but the conjunctival infection is asymptomatic.     

Protection from challenge following administration of a canarypox virus-vectored recombinant feline leukemia virus vaccine in cats previously vaccinated with a killed virus vaccine

OBJECTIVE: To compare protection against FeLV challenge obtained following administration of 2 doses of an adjuvanted, chemically inactivated, whole FeLV (FeLV-k) vaccine with protection obtained following administration of 1 dose of an FeLV-k vaccine followed by 1 dose of a canarypox virus-vectored recombinant FeLV (rCP-FeLV) vaccine. DESIGN: Prospective study. ANIMALS: Thirty-two 9-week-old domestic shorthair cats. PROCEDURE: Cats received 2 doses of the FeLV-k vaccine SC, 21 days apart (n = 11); 1 dose of the FeLV-k vaccine SC and, 21 days later, 1 dose of the rCP-FeLV vaccine transdermally (11); or 2 doses of physiologic saline (0.9% NaCl) solution (control; 10). Four weeks after the second vaccine dose, all cats were challenged with FeLV by means of oronasal administration. Blood samples were collected at weekly intervals beginning 21 days after challenge, and serum was tested for FeLV antigen. RESULTS: All 10 control cats became persistently infected (ie, FeLV antigen detected in > or = 3 consecutive serum samples) following FeLV challenge, whereas only 1 of 11 cats that received 2 doses of the FeLV-k vaccine and none of the 11 cats that received 1 dose of the FeLV-k vaccine and 1 dose of the rCP-FeLV vaccine did. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that protection against FeLV challenge obtained following SC administration of a single dose of an FeLV-k vaccine followed, 21 days later, by transdermal administration of a single dose of an rCP-FeLV vaccine was similar to that obtained following SC administration of 2 doses of the FeLV-k vaccine 21 days apart.     

Inactivated suckling mouse brain rabies vaccine provides short-term immunity in capuchin monkeys (Cebus apella).

Eight capuchin monkeys (Cebus apella) were vaccinated against rabies with an inactivated suckling mouse brain vaccine (SMBV). Three 1-ml doses of 2% brain tissue suspension were given by i.m. injection at 0, 30, and 60 days. Blood samples were collected at 0, 30, 60, 90, 150, 210, 240, 300, and 365 days and were tested by simplified fluorescence inhibition to titer-neutralizing antibodies. All of the animals developed neutralizing antibodies with titers >0.5 IU/ml after vaccination, but the immune response persisted for only 122.3 +/- 32.6 days. The SMBV was able to induce immune response in the capuchin monkeys, but protection was short-lived.     

以小鼠为模型评价重组伪狂犬病病毒rPRV-HA的免疫效力

以小鼠为动物模型，对此前构建的表达H3N2亚型猪流感病毒（SIV）血凝素（HA）基因的重组伪狂犬病病毒（rPRV-HA）进行了免疫效力评价。按每只10^5.0 TCID50 rPRV-HA的剂量通过滴鼻接种8周龄雌性BALB／c小鼠（n=60），同时设Bartha-K61免疫对照组（n=60）、非免疫攻毒对照组（n=20）和非免疫不攻毒对照组（n=10）。于免疫后不同时间分别从rPRV-HA免疫组和Bartha—K61免疫对照组随机剖杀一定数量的小鼠，其余小鼠于免疫后第28天用10^5.0 TCID50同亚型SIV毒株A／Swine／Heilongjiang／74／2000（H3N2）进行强毒攻击。攻毒后第4、7、14天分别剖杀小鼠，进行间接免疫荧光、病毒分离、血清学和病理组织学检测。结果表明，重组病毒主要分布于肺脏；免疫后14d起，从rPRV—HA免疫组及Bartha—K61免疫对照组均可检测到针对PRV的荧光抗体；从rPRV—HA免疫组可以检测到针对SIV的荧光抗体和血凝抑制抗体，而各对照组均呈阴性。攻毒后从rPRV—HA免疫组小鼠未分离到攻击病毒，血凝抑制抗体显著升高，病理变化显著轻于对照组，表明rPRV—HA免疫小鼠可以抵抗同亚型SIV的攻击，可以作为rPRV—HA免疫效力评价模型。     

Experimental enteric infection of gnotobiotic piglets with Chlamydia suis strain S45

Enteric chlamydial infections of pigs with Chlamydia (C.) suis are frequent and often subclinical. The enteric pathogenicity of C. suis strain S45 was investigated in gnotobiotic piglets. Piglets from three litters (n=31) were inoculated with egg-grown chlamydiae at 2-3 days of age (n=17) or used as controls (n=14). They were observed for clinical signs, killed and necropsied sequentially at 2-13 days postinoculation (DPI). Feces were collected daily and investigated with an ELISA for chlamydial antigen. At necropsy, specimens were collected for histopathology and for immunohistochemical, PCR-based, and serological (complement fixation test, ELISA) detection of chlamydiae. Chlamydial replication and associated symptoms and lesions were observed from 2 to 13 DPI and were particularly pronounced within the first week PI. Clinical symptoms consisted of moderate-to-severe diarrhea, slight and transient anorexia, weakness and body weight loss. Immunohistochemistry and ELISA revealed that chlamydial replication was particularly marked at 2-4 DPI and primarily located in the small intestinal villus enterocytes. Further sites of replication included large intestinal enterocytes, the lamina propria and Tunica submucosa, and the mesenteric lymphnodes. Histopathological changes included moderate-to-severe villus atrophy with flattened enterocytes and focal villus tip erosions, and moderate mucosal inflammatory cell infiltrates and lymphangitis in the small intestine. PCR of spleen tissue and blood was mostly negative for chlamydiae, indicating that they did not substantially disseminate into the host up to 13 DPI. All sera were negative for anti-chlamydial antibodies. In conclusion, C. suis strain S45 elicited significant enteric disease and lesions in gnotobiotic piglets indicating its pathogenic potential for swine.     

Effect of vaccination with a modified-live porcine reproductive and respiratory syndrome virus vaccine on dynamics of homologous viral infection in pigs

OBJECTIVE: To determine effects of vaccination protocols with modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine on persistence and transmission of virus in pigs infected with a homologous isolate and determine clinical and virologic responses following heterologous viral challenge. ANIMALS: Four hundred forty 6- to 8-week-old PRRSV-na?ve pigs. PROCEDURES: Pigs were allocated into 5 groups. Groups A to D were inoculated with wild-type PRRSV VR2332. Group A (positive control pigs) received PRRSV only. Groups B, C, and D received modified-live PRRSV vaccine (1, 2, or 3 doses). Group E served as a negative control group. To evaluate viral transmission, sentinel pigs were introduced into each group at intervals from 37 to 67, 67 to 97, and 97 to 127 days postinoculation (DPI). To evaluate persistence, pigs were euthanized at 37, 67, 97, or 127 DPI. To assess clinical and virologic response after challenge, selected pigs from each group were inoculated at 98 DPI with a heterologous isolate (PRRSV MN-184). RESULTS: Mass vaccination significantly reduced the number of persistently infected pigs at 127 DPI. Vaccination did not eliminate wild-type PRRSV; administration of 2 or 3 doses of modified-live virus vaccine reduced viral shedding after 97 DPI. Previous exposure to wild-type and vaccine virus reduced clinical signs and enhanced growth following heterologous challenge but did not prevent infection. CONCLUSIONS AND CLINICAL RELEVANCE: Findings suggest that therapeutic vaccination may help to reduce economic losses of PRRSV caused by infection; further studies to define the role of modified-live virus vaccines in control-eradication programs are needed.     

Effect of levamisole on parenteral vaccines for swine dysentery

Leucocyte migration-inhibition and humoral antibody responses (HAR) were demonstrated in swine immunized with particulate or soluble antigen of Treponema hyodysenteriae. Levamisole (at the recommended deworming level), given simultaneously with particulate vaccine did not significantly (P greater than 0.05) enhance nor suppress the leucocyte migration-inhibition response. However, an enhancing leucocyte migration-inhibition of the drug was seen in pigs given soluble vaccine in combination with levamisole compared with those receiving soluble antigen alone. Levamisole generally suppressed the HAR throughout the immunization schedule of pigs given soluble or particulate vaccine. There was no significant suppression (P greater than 0.05) of clinical signs of swine dysentery (SD) in animals given particulate vaccine nor in those receiving this vaccine plus levamisole. However, pigs receiving soluble vaccine plus levamisole had fewer clinical signs of SD as well as significantly (P less than 0.05) fewer shedding episodes of T. hyodysenteriae than those given soluble antigen alone. When compared with the control pigs, swine vaccinated with soluble antigen had fewer days of total diarrhoea and shedding episodes of T. hyodysenteriae. However, the diarrhoea of vaccinated pigs was not significantly (P greater than 0.05) delayed when compared to the unvaccinated swine.     

猪囊虫病基因工程疫苗的研制及应用

从猪带绦虫六钩蚴cDNA文库中筛选目的基因并进行克隆表达，重组抗原用血清学方法和猪体免疫试验进行鉴定，筛选保护性抗原基因。将具有免疫保护作用的重组抗原纯化，并与免疫刺激复合物佐剂结合，制成猪囊虫病基因工程疫苗。对工程菌的培养发酵、重组抗原的下游纯化及疫苗的配制、疫苗的中间试制进行了研究，确定了一套比较完善的生产工艺。应用昆明小鼠建立了猪囊虫病的实验动物模型，应用该模型进行上述疫苗的免疫预防试验，免疫1次和2次的免疫保护率分别为96．9％和98．4％。本动物的免疫预防试验显示，疫苗安全性好，免疫保护率达92．2％，且免疫组发现的囊虫多数已死亡。在流行区进行了田间和区域试验。免疫猪未有不良反应，囊虫感染率分别由20％降为1．1％和5．4％降为0．21％。用该疫苗对人工感染的囊虫病猪进行免疫治疗，发现在感染早期的治疗效果较好。免疫动物的体液免疫和细胞免疫的检测结果显示，该疫苗刺激机体产生抗体的时间早、持续时间长、效价高；可显著提高淋巴细胞转化率及E－RFC和Ea-RFC细胞、ANAE^ 细胞和粗粒型ANAE^ 细胞、抑制／杀伤性T细胞亚群的数量。以上结果表明，用大肠杆菌表达的重组抗原制备的猪囊虫病基因工程疫苗安全、高效，且成本低廉，可规模化生产，有成为预防猪囊虫病的一种新型生物制剂。     

Prevalence of Arkansas-type infectious bronchitis virus in Delmarva peninsula chickens

The prevalence of Arkansas (Ark)-type infectious bronchitis virus (IBV) in Delmarva peninsula broiler-type chickens was determined. The immunity of 5-to-11-week-old commercial broilers was evaluated by intraocular inoculation with Ark-type DPI strain (Ark DPI) challenge virus and collection of tracheal swabbings 5 days later. Serum Ark-type antibody titers were obtained using the virus-neutralization test. Eighty-five flocks were tested from January to August 1981. Nearly 60% of the flocks had substantial (greater than or equal to 70%) local immunity of the upper respiratory tract. Twenty-two percent had intermediate (50-69%) and 19% of the flocks had low (less than or equal to 40%) levels of local immunity. Serum antibody titers generally agreed with challenge results. In addition, high Ark-type IBV neutralizing-antibody titers were found in 16 Delmarva broiler breeder flocks. Seven current IBV field isolates were characterized for antigenic similarity to Ark DPI. Four isolates contained Ark antigen(s) based on significant neutralization in virus-neutralization tests and on substantial immunity to challenge afforded by Ark DPI virus immunization. Three isolates did not appear to contain Ark antigen(s). Immunization of chickens with Ark DPI virus afforded substantial protection against Connecticut- and homologous-type virus challenge, partial immunity (63%) against JMK, and no protection against the Massachusetts 41 strain of IBV.     

Tissue reaction and immunity in swine immunized with Actinobacillus pleuropneumoniae vaccines.

These studies were done to develop a subunit vaccine for swine that would protect against disease, but not create unacceptable tissue reactions at the immunization site. Swine were used to evaluate the local effects of subunit vaccines prepared from extracts of Actinobacillus pleuropneumoniae serotype 1 containing one of a wide variety of adjuvants. The antigen was an anionic fraction of a saline extract of A. pleuropneumoniae (ANEX). The adjuvants used were vegetable oils (peanut, sesame, canola, or corn oils, vitamin E, or Lipposyn II emulsion); mineral oil (Marcol-52) and other materials (aluminum hydroxide, polyethylene glycol, Quil-A, Amphigen, or Emulsigen-Plus). Two types of experiments were done. In the 1st set of experiments, pigs were given multiple simultaneous injections in different sites and euthanized on days 1, 3, 7, 14, 21, or 28. Tissues were examined for gross and histopathological lesions. In the 2nd set of experiments, 48 pigs were allocated to 6 groups and vaccinated twice with a vaccine containing ANEX antigen combined with one of various adjuvants. Antibody responses and protection from challenge were evaluated. Among the adjuvants that were tested, mineral oils induced protective immunity, although the mineral oil Marcol-52 resulted in severe tissue reactions. The vegetable oils induced little protective immunity, and some of them were quite irritating. The response to the other materials ranged from little irritation or protection induced by the vaccine containing aluminum hydroxide to effective protection without irritation after vaccination with ANEX/Amphigen or ANEX/Emulsigen-Plus combinations. In conclusion, swine were protected against disease by a subunit vaccine that did not create unacceptable tissue reaction at the immunization site.     

Immunization of cattle against Aujeszky's disease with inactivated adjuvant vaccine

The experiments with sheep and young cattle were carried out to test the immunizing efficacy of inactivated adjuvant vaccine against Aujeszky's disease. The vaccine application at doses of 1 ml and 2 ml to lambs at the age of eight to ten months caused the neutralizing antibody production with a significant rise of titres after revaccination. A survival of infection induced with a dose of 10(5.5) TKID50 of virulent virus was recorded in 62.5% of once vaccinated animals and in 87.5% of twice vaccinated animals. When applying different doses of vaccines (from 1 to 10 ml) to young cattle, the antibody reaction level was directly dependent on the inoculum quantity. The double inoculation of animals with vaccines of 2 ml and 5 ml caused the neutralizing antibody production at titres of 1:35, or 1:46. The animals, immunized with the live or inactivated IBR-vaccine possessing high antibody titres against IBR-virus, reacted upon the vaccination with inactivated Aujeszky's vaccine anamnestically, by early production of antibodies in high titres. Metaphylactic vaccination (2 ml of vaccine) of cattle in herds with an acute course days, however earlier during five days from the revaccination when it was carried out in seven days following the first vaccination.     

Comparison of two swine Mycoplasma hyopneumoniae enzyme-linked immunosorbent assays for detection of antibodies from vaccinated pigs and field serum samples.

Mycoplasma hyopneumoniae (Mhyo) causes mycoplasmal pneumonia, an economically important disease of swine. Serodiagnosis of Mhyo is based on the current available commercial enzyme immunoassays for detection of swine antibodies against Mhyo, which are the indirect enzyme-linked immunosorbent assay (ELISA) and the blocking ELISA (B-ELISA). Because of the limited information available for these ELISAs, these 2 assays were compared by testing 347 serum samples collected from vaccinated pigs at 0, 13, 28, 43, and 62 days postimmunization (DPI), 50 samples from nonvaccinated pigs, and 1,013 field serum samples. The results of comparison study showed that the specificity for both ELISAs was 99.2% generated from 139 non-vaccinated negative samples. The sensitivities for indirect ELISA generated from samples collected from animals that received the vaccine at DPI 13, 28, 43, and 62 were 0%, 95.7%, 88.4%, and 92.6%, respectively, whereas the sensitivities for B-ELISA were 0%, 98%, 100%, and 97%, respectively. The overall agreement of 96.7% and 80.3% was generated between 2 ELISAs from negative and vaccinated pigs and from field samples, respectively.     

Ascaris suum--vaccination of mice with liposome encapsulated antigen.

Vaccination with liposome encapsulated adult crude antigen with and without coencapsulated immunomodulator (levamisole) in a mice/larval Ascaris suum model provided protection against a challenge infection (2000 eggs) in mice immunised by immobilised antigen. The best results (88.9% protection) were obtained with a combination of two doses of liposome entrapped antigen with leamisole. Vaccination with liposome vaccine without modulator was slightly less effective (78.7% protection). A single dose of vaccine was ineffective (14.3% protection). Application of the soluble antigen without any adjuvants led to the enhancement of worm yield in lungs and liver.     

Comparison of the safety and efficacy of a recombinant feline leukemia virus (FeLV) vaccine delivered transdermally and an inactivated FeLV vaccine delivered subcutaneously

The efficacy of a new recombinant FeLV vaccine (rFeLV), delivered transdermally via a needle-free delivery device was compared to that of an inactivated FeLV vaccine (FeLV-k), administered subcutaneously, with a conventional needle and syringe. Kittens were immunized with either rFeLV (0.25 ml, transdermal) or FeLV-k (1 ml, subcutaneous); or they were sham-vaccinated with physiologic saline (0.25 ml, transdermal). Two vaccinations were administered 21 days apart. Injection sites were monitored for any acute or subacute reactions relative to vaccine administration. Four weeks following the final vaccination, all cats were subject to oro-nasal FeLV challenge. Blood was collected for determination of FeLV antigenemia (p27) at weekly intervals beginning three weeks post-challenge. All of the vaccinated cats from both groups resisted FeLV challenge; and 90% of the control cats developed persistent FeLV antigenemia in response to challenge. No acute or persistent injection site reactions were observed. The rFeLV, delivered transdermally, provides protection against persistent FeLV antigenemia following a robust challenge that is equivalent to that of FeLV-k.     

Evaluation of a protective immunity induced by an inactivated influenza H3N2 vaccine after an intratracheal challenge of pigs.

A challenge study was conducted to evaluate the safety and efficacy of an inactivated influenza H3N2 virus vaccine combined with Quil A/Alhydrogel mixture under controlled conditions in piglets. Twenty-four piglets from 12 sows were allocated to 2 groups; injected intramuscularly with 2 doses of the tested vaccine or with PBS at 2 wk intervals and challenged intratracheally with 105TCID50 of the H3N2 swine influenza virus 6 d after the 2nd immunization. Clinical and virological parameters were recorded for 4 d after the challenge. The use of the tested vaccine produced high serum hemagglutination-inhibition titers against the swine H3N2 strain virus. This strong immune response suppressed all clinical signs and viral shedding and reduced pulmonary lesions due to the challenge in the vaccinated group, without causing any secondary effects. Our results suggest that the serum HI titers correlated with the degree of protection induced by an inactivated swine influenza H3N2 vaccine.