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.     

Immune responses of breeding chickens to trivalent oil emulsion vaccines: responses to Newcastle disease and infectious bursal disease

Bivalent Newcastle disease (ND)/infectious bursal disease (IBD) and trivalent ND/IBD/infectious bronchitis (IB) inactivated oil emulsion vaccines were prepared in the laboratory and evaluated under field conditions. Broiler breeder parent chickens previously vaccinated with live vaccines were inoculated with commercial monovalent ND and experimental bivalent or trivalent oil emulsion vaccines. The commercial vaccine induced a higher initial ND haemagglutination inhibition (HI) response than the experimental vaccines but, by 34 weeks after vaccination, the mean ND HI levels were not significantly different in any of the three flocks. All three vaccines provided sufficient ND immunity to protect against the clinical disease and egg production losses. The IBD responses of both flocks vaccinated with oil emulsion vaccine were similar to each other and only slightly lower than those flocks vaccinated with monovalent IBD oil emulsion vaccine in earlier experiments. Six weeks after vaccination, sufficient immunity was transferred to protect all the progeny against IBD challenge up to 33 days of age and some of them up to 45 days of age. Thirty-four weeks after vaccination of the parents with oil emulsion vaccine, the progeny were totally immune up to 27 days of age and some of them were immune until 37 days. Application of oil emulsion vaccines in bivalent or trivalent form did not impair the responses of the chickens to the monovalent components.     

Efficacy of experimental Newcastle disease water-in-oil oil-emulsion vaccines formulated from squalane and squalene

Water-in-oil inactivated Newcastle disease oil-emulsion vaccines were formulated with the terpene oils squalane or squalene, or mixtures thereof, and injected into 4-week-old broilers. Vaccine efficacy based on hemagglutination-inhibition (HI) titers was comparable to that of control mineral oil vaccines. Tissue reaction to intramuscular injection of the terpene oil emulsion vaccines was greatly reduced 3 weeks post-vaccination compared with that of mineral oil-based vaccine. Viscosity of the terpene oil vaccines was satisfactory but increased three to four times that of mineral oil vaccine when the antigen phase volume increased from 5% to 20%.     

Molecular identification of the vaccine strain from the inactivated oil emulsion H9N2 low pathogenic avian influenza vaccine

In order to control the H9N2 subtype low pathogenic avian influenza (LPAI), an inactivated vaccine has been used in Korea since 2007. The Korean veterinary authority permitted the use of a single H9N2 LPAI vaccine strain to simplify the evolution of the circulating virus due to the immune pressure caused by the vaccine use. It is therefore important to determine the suitability of the vaccine strain in the final inactivated oil emulsion LPAI vaccine. In this study, we applied molecular rather than biological methods to verify the suitability of the vaccine strain used in commercial vaccines and successfully identified the strain by comparing the nucleotide sequences of the hemagglutinin and neuraminidase genes with that of the permitted Korean LPAI vaccine strain. It is thought that the method used in this study might be successfully applied to other viral genes of the LPAI vaccine strain and perhaps to other veterinary oil emulsion vaccines.     

Use of infectious bursal disease vaccines in chicks with maternally derived antibodies

Day-old chicks with maternally derived infectious bursal disease (IBD) antibodies were inoculated with IBD oil emulsion vaccine. The vaccine protected at least 85 per cent of the chicks when they were challenged at four weeks old with virulent IBD virus and at least 90 per cent of those challenged at seven weeks of age. There was no benefit in using a combination of the oil emulsion vaccine and a live IBD vaccine.     

Protection of laying hens against infectious bronchitis with inactivated emulsion vaccines

Commercially-reared laying chickens were challenged at 31 weeks of age with a virulent infectious bronchitis (IB) virus. They showed a sharp drop in egg production, despite having been vaccinated at four and eight weeks old with live attenuated IB vaccines to a recommended schedule. In contrast, similar birds that had been further immunised at point-of-lay with inactivated oil emulsion IB vaccine, or with a combined IB/Newcastle disease (ND) emulsion vaccine, showed no detectable fall in egg production after the same challenge. Unvaccinated susceptible specific pathogen-free birds challenged at the same time stopped laying almost completely. In the birds revaccinated with emulsion vaccine, measurement of haemagglutination inhibition antibody levels to IB showed their geometric mean titres to be raised from less than 5 log2 at the time of vaccination to over 10 log2 four weeks later. Their antibody levels did not rise further followining the IB challenge whereas in the birds that had not been revaccinated antibody rises to nearly 10 log2 were detected after the same challenge. For pullets vaccinated earlier with live IB vaccine, revaccination with inactivated IB or IB/ND oil emulsion vaccine at point-of-lay provides a safe and effective way of protecting their egg production against IB infection.     

鸭疫里默氏杆菌多价灭活苗研制的研究报告

用三个血清型的菌株,分别是JYL-1、JYL-2、JYL-7,用两种佐剂,分别是蜂胶和油乳剂共制备了两种多价菌苗,即JYL-1、JYL-2、JYL-7（1、2、7三个血清型）三个RA血清型混合多价蜂胶苗和油乳剂苗。试验结果表明。蜂胶复合佐剂多价苗具有产生免疫保护速度快,免疫持续时间长的优点,接种后第3天即产生部分免疫保护力,第120天时仍具有完全保护力;油乳剂多价苗产生保护力的速度较慢,接种后第10天时开始表现出部分免疫保护,其完全保护力也可持续到接种后第120天。免疫后13d时,免疫保护率可达90％左右。免疫后16d时保护率为100％。免疫后120d时,两种多价菌苗的免疫保护率均可达到100％,免疫后150～180d时,免疫保护率可达800左右。统计学分析证明,菌苗的安全性良好。用3个免疫剂量的菌苗所做的安全试验表明无明显不良反应,菌苗在4℃保存一年,18～22℃室温保存4个月不影响菌苗的免疫效力。田间试验表明,用菌苗免疫鸭群后,可有效地使鸭群抵抗鸭疫里默氏杆菌的感染。     

Guinea pig protection test as indicator of potency of oil emulsion foot-and-mouth disease vaccines

A vaccine potency test is described involving virus challenge to six groups of 10 guinea pigs at five weeks after vaccination. Sixteen oil emulsion foot-and-mouth disease vaccines were so tested and nine retested after storage at 4 degrees C for up to 28.3 months. The results were compared with those of the routinely used oil emulsion vaccine potency test (protection afforded to eight pigs challenged 21 days after vaccination). When guinea pig estimates of 3 log2 PD50 or more were obtained, then, with one exception, the batches protected all or almost all pigs from challenge, but when the guinea pig estimates were less than 1 log2 PD50, the vaccines failed to protect five out of eight pigs. The sensitivity and reproducibility of the guinea pig method, established by repeated tests on two vaccine batches, seemed acceptable. The results suggested that guinea pig estimates might provide a suitable substitute for pig challenge potency tests because they reflected the potency of the vaccines, were likely to involve smaller standard errors and caused less discomfort to animals.     

Comparison of alum-absorbed or non-alum-absorbed oil emulsion vaccines containing either pilate or non-pilate Bacteroides nodosus cells in inducing and maintaining resistance of sheep to experimental foot rot

Highly pilate (P) or non-pilate (NP) cells of Bacteroides nodosus were compounded into oil emulsion (O) either with or without prior absorption onto alum (A). The abilities of these four preparations (referred to as PAO, NPAO, PO and NPO vaccines) to stimulate antibody production and to protect sheep from foot rot were compared. Two injections of PAO vaccine protected sheep against homologous challenge 12 weeks after the second dose by PO, NPO and NPAO vaccines were less effective. Sheep were protected against homologous challenge for 14 weeks after a single dose of PAO vaccine and for 22 weeks after three doses; an ameliorative effect was still evident 40 weeks after the third dose. Protection against challenge with two heterologous strains was demonstrated at six weeks after three doses of vaccine. A numerical system of scoring the lesions also confirmed that foot rot in vaccinated sheep challenged outside the 'protective' period of the vaccine was somewhat less severe than in controls. PAO vaccine induced much higher and more persistent titres of agglutinins than the other vaccines tested. There was a relationship between agglutinin titres and resistance to homologous challenge.     

鸽新城疫油乳剂灭活苗的研制

本文报导了一种源自病鸽的鸽新城疫病毒ＱＬ株制备的灭活油乳剂疫苗。以此种疫苗接种后,接种鸽均正常无任何病症,接种疫苗获得保护能抵抗鸽新城疫病毒株攻击的鸽子数多于接种ＬａＳｏｔａ油乳剂疫苗的鸽子。已在野外以此种疫苗接种了大量的鸽子,效果满意     

THE SEROLOGICAL RESPONSE OF CHICKENS TO OIL EMULSION VACCINES CONTAINING THE V4 STRAIN OF NEWCASTLE DISEASE VIRUS

SUMMARY Experiments were conducted with vaccines containing the V₄ strain of Newcastle disease virus (NDV). Both living aqueous vaccines and vaccines consisting of virus incorporated in an oil emulsion were used. The calculated dose of virus contained in the oil emulsion vaccine was 10^8,7 50% embryo infectious doses (EID₅₀) per bird dose. Haemagglutinin inhibition (HI) antibody levels of 8 are presumed protective. One-day-old chicks with low levels of maternal antibody were vaccinated intraocularly with 10^6,3EID₅₀ of live vaccine, and concurrently with oil emulsion vaccine. Presumed protective levels of antibody were present at two weeks post vaccination and were maintained for at least seven weeks longer. When adult birds 15 weeks old with no previous exposure to NDV were vaccinated intraocularly with 10^6,7EID₅₀ per bird, protective levels of antibody were produced within a week. Unvaccinated birds put in contact with the vaccinated birds produced similar antibody levels within 14 days. Revaccination with oil emulsion vaccine after antibody levels had fallen resulted in a rapid response with high levels of antibody. When antibody-free adult commercial birds with an unknown history of exposure to NDV were vaccinated intramuscularly with oil emulsion vaccine, high antibody levels were produced for at least 21 weeks. Concurrent intraocular inoculation with 10^7,0EID₅₀ live virus did not enhance the response. Natural infection of unvaccinated birds occurred during the experiment. This was detected by the presence of HI antibody levels of short duration. When antibody-free commercial birds were inoculated intramuscularly with oil emulsion vaccine containing 10^6,0, 10^7,0, or 10^8,0EID₅₀ per bird dose, 100% of birds inoculated with the highest dose produced presumed protective levels of antibody within two weeks, as compared with a 5-week delay when using the 10^7,0EID₅₀ per bird dose.     

Efficacy of avridine as an adjuvant for Newcastle disease virus antigen in chickens

Avridine, a lipoidal amine with interferon-inducing and adjuvant properties, was an effective adjuvant for Newcastle disease antigen (NDA) in chickens. Eleven vaccine lots were evaluated: 2 commercial water-in-oil vaccines, 4 experimental oil emulsion vaccines, 4 avridine-containing vaccines, and a control lot of nonadjuvanted antigen. Avridine significantly enhanced the immunologic responses of chickens against NDA. Chickens vaccinated with the avridine-containing vaccines had significantly higher antibody titers (hemagglutination inhibition) than did chickens vaccinated with the commercial vaccines. Experimental oil emulsion vaccines prepared from the same antigens as avridine-adjuvanted vaccines induced higher hemagglutination inhibition antibody titers after primary but not after booster vaccination. Use of avridine as an adjuvant for NDA in vaccines for chickens induced immunologic protection rates similar to those induced by oil emulsion vaccines, without causing the reactogenic and tissue residue problems associated with the use of oil vaccines in chickens.     

新城疫病毒接种鸡血清中IgM、IgG的动态变化

将１６只ＳＰＦ鸡分成４组，每组４只，分别接种新城疫油乳剂苗，新城疫、传染性支气管炎、传染性法氏囊病三联油乳剂苗，ＬａＳｏｔａ弱毒苗和Ｍｕｋｔｅｓｗｅｒ苗。接种后每隔２ｄ采血１次，用ＥＬＩＳＡ测定新城疫特异性ＩｇＭ和ＩｇＧ抗体。结果：油乳剂灭活苗接种组均无特异性ＩｇＭ抗体出现，特异性ＩｇＧ抗体高峰期出现在接种后２２ｄ，ＬａＳｏｔａ弱毒苗和Ｍｕｋｔｅｓｗｅｒ苗接种组均有特异性ＩｇＭ抗体出现，高峰期为接种后第９ｄ。各接种组经强毒攻击后，均出现ＩｇＭ反应，ＩｂＧ呈典型的回忆反应。进一步试验用ＬａＳｏｔａ灭活苗经肌肉、静脉接种和加氢氧化铝胶佐剂后肌肉接种，经测定，接种鸡均无或仅有很低水平的特异性ＩｇＭ抗体产生。     

Protection of chickens against a lethal challenge of Escherichia coli by a vaccine containing CpG oligodeoxynucleotides as an adjuvant

Synthetic oligodeoxynucleotides (ODN) containing cytosine-phosphodiester-guanine (CpG) motifs (CpG-ODN) have been shown to be effective immunoprotective agents and vaccine adjuvants in a variety of bacterial, viral, and protozoan diseases in different animal species. The objective of this study was to compare the immune response of chickens to a killed Escherichia coli vaccine combined with oil in water emulsion or with CpG-ODN. Birds were vaccinated with killed E. coli antigens with either 10 or 50 microg of CpG-ODN on days 10 and 20 of age. At day 30, a virulent isolate of homologous E. coli was applied on a scratch site on the caudal abdominal region. Birds were examined for 10 days post-E. coli challenge, and pathologic and bacteriologic assessments were conducted on all birds that were either found dead or euthanized. The E. coli vaccine group that received no CpG-ODN had a survival rate of 65%. In contrast, groups that received the vaccine with CpG-ODN adjuvant had significantly higher survival rate of 92% (P < 0.01) with isolation of low numbers of E. coli from internal organs. Total IgG against E. coli antigens was highest in groups that received CpG-ODN as an adjuvant. Birds that received vaccine containing CpG-ODN had minimal inflammatory reaction without tissue necrosis at the injection site. Severe tissue necrosis was present in birds that received vaccine containing oil in water emulsion adjuvant. This study demonstrated that CpG-ODN is an effective vaccine adjuvant in chickens and results in minimal tissue destruction. This study is the first study in which CpG-ODN has been demonstrated to produce an adaptive immune response, at a significant level, against an extracellular bacterial infection in chickens.     

益生菌基因组DNA作为疫苗佐剂对鸡免疫效果的影响

一次试验选用1株益生菌的基因组作为禽流感病毒(AIV)H5N1亚型灭活油乳苗的佐剂,设不同剂量,分口服和同位点注射佐剂、疫苗的途径免疫鸡群;二次试验扩大菌株类型,以5株益生菌的基因组作为佐剂同位点注射鸡群,定期检测特异性抗体水平。一次试验显示在所有检测时间点上,疫苗对照组的抗体效价高于(P>0.05)或者显著高于(P<0.05)益生菌基因组的,且注射佐剂组的抗体效价高于口服佐剂的效价;二次试验显示疫苗对照组的抗体效价在前期高于(P>0.05)或者显著高于(P<0.05)益生菌基因组的效价,后期略低于某些益生菌组的(P>0.05),且各种益生菌基因组不同程度的推迟了抗体高峰期。     

Application of diethylstilbestrol dipropionate in bulls. I. Excretion of residues in urine and faeces and histological and immunohistochemical changes in the prostate

In this experiment 20 one year old bulls received a single intramuscular injection of the anabolic preparation diethylstilbestrol dipropionate (DES-DP) (an oil preparation or an emulsion). Four animals received a corresponding placebo. The application of DES-DP to bulls caused characteristic histological alterations in the peripheral glandular epithelium of the prostate, which could be observed until four weeks after treatment. The value of histological investigation as a screening method was, however, limited by the occurrence of only few metaplastic lesions and a rapid recovery. By contrast, immunohistochemistry using a polyclonal cytokeratin antiserum K40 appeared to be a specific and very sensitive method to detect oestrogen-induced lesions in the prostate. In only two animals, six weeks after injection with the DES-emulsion, false-negative results were obtained, demonstrating the potential value of this screening method. The excretion of DES in the urine and faeces was monitored using radioimmunoassay following chromatographic purification of the urine and faeces extracts. The excretion of DES in urine was faster for animals of the oil group. The DES content in urine decreased to the 1 microgram/l level after 42 days (emulsion group) or 70 days (oil group). The excretion in faeces was comparable to that in urine. After day 21 the excretion patterns of the two excreta were indistinguishable.     

Immunoadjuvant effects of bacterial genomic DNA and CpG oligodeoxynucleotides on avian influenza virus subtype H5N1 inactivated oil emulsion vaccine in chicken

This study investigated the immunoadjuvant effects of three types of bacterial genomic DNA and CpG oligonucleotides (CpG ODN) on the avian influenza virus (AIV) subtype H5N1 inactivated oil emulsion vaccine under two immunization strategies. The genomic DNA extracted from Escherichia coli O₂, Staphylococcus aureus, Streptococcus faecalis FQ68, and synthetic CpG ODN were used as adjuvants, and their effects on the AIV oil emulsion vaccine were examined in chickens. The results indicated that when administered separately from the vaccine, adjuvants induced lower haemagglutination inhibition (HI) titres and serum IgG titres but resulted in higher concentrations of IFN-γ and IL-10. In contrast, when combined with the oil emulsion vaccine prior to inoculation, CpG ODN induced higher HI, IgG titres and IFN-γ concentration but resulted in lower IL-10 concentration. These data suggest that, depending on the immunization approaches, adjuvants may exert distinct immune effects in chickens receiving AIV H5N1 oil emulsion vaccine: the prior incorporation of CpG ODN into the vaccine may augment both the humoral and Th1 type immune responses, while separate inoculation of adjuvants has not shown better adjuvanticity.     

鸡败血支原体接种鸡体液免疫的变化规律

SPF鸡接种鸡败血支原体R株、F株和油乳剂苗后，用ELISA检测IgM、IgG和IgA的变化规律，发现特异性IgM出现时间存在差异；强毒株感染后IgM始终存在，因此可作为临诊感染的指证。油乳剂苗能产生IgA，证明油乳剂苗能诱导产生局部免疫。     

Effect of adjuvants on antibody responses of sheep immunised with recombinant pili from Dichelobacter nodosus

Objective To compare the effects of two oil emulsion adjuvants (incomplete Freunds adjuvant and a proprietary oil adjuvant), DEAE-dextran, L-tyrosine particles and Quil A on the humoral immune responses of sheep immunised with recombinant pili of Dichelobacter nodosus (strain A).
Procedure Antibody titres were studied for up to 32 weeks and were measured by bacterial agglutination and ELISA. The relative avidity of antibodies for pili was determined and the incidence and severity of adverse reactions at the site of injection of vaccines were recorded.
Results The oil emulsion adjuvants and Quil A were more effective than either DEAE-dextran or L-tyrosine at stimulating antibodies in sheep. The incidence and severity of adverse reactions was lower in sheep which received vaccines containing either Quil A or DEAE-dextran than in sheep which received vaccines containing oil emulsion adjuvants. L-tyrosine had no adverse effects.
Conclusion Quil A was as effective as oil adjuvants at stimulating high levels of antibodies against recombinant pili in sheep and had the significant advantage of being less irritant after subcutaneous injection.     

Cell mediated and humoral immune response of chickens to inactivated oil-emulsion infectious bronchitis vaccine

A lymphocyte transformation microassay was used to study cell mediated immunity (CMI) in chickens following primary and secondary vaccination with inactivated oil emulsion infectious bronchitis (IB) vaccine and subsequent challenge with Massachusetts-41 (M-41). Humoral immunity was monitored for comparison, using the haemagglutination inhibition (HI) microassay. Positive stimulation indices (2 to 2.7 after primary and 2 to 4.8 after secondary vaccination) were lower and HI titres were higher than those previously reported following primary and secondary vaccination with live IB vaccines. The highest HI titres appeared in birds which had received the inactivated vaccine as a secondary vaccination. Challenge of vaccinated and revaccinated birds resulted in strong HI and weak CMI secondary responses. There was no correlation between CMI and HI antibody production. Monitoring egg production and clinical signs showed that a high level of protection against challenge resulted from revaccination with an inactivated oil adjuvant vaccine.