Field trials with a bivalent vaccine (HVT and SB-1) against Marek's disease

White leghorn chickens on five farms were given a bivalent Marek's disease (MD) vaccine consisting of turkey herpesvirus (HVT) and SB-1 (a nononcogenic MD virus); other chickens received only HVT. The farms had histories of "vaccination failures," presumably owing to an exceptionally virulent challenge MD virus. The bivalent vaccine uniformly protected chickens better than HVT alone between 12 and 16-20 weeks of age, when serious MD losses occurred. During that period, total mortality in groups given both viruses ranged from 0.39 to 1.26% (mean 0.86%), whereas that in HVT-vaccinated groups not exposed to SB-1 varied from 1.92 to 7.44% (mean 3.43%). Chickens in pens or rows with close contact to those given bivalent vaccine also had low MD mortality rates (0.46-1.06%, mean 0.77%), probably from the spread of SB-1.     

Isolation of very virulent Marek''s disease viruses from vaccinated chickens in Australia

Very virulent Marek's disease viruses (vvMDV), defined as isolates against which the herpesvirus of turkey (HVT) vaccine provide poor protection, have been isolated from poultry flocks in both the United States and Europe. Twenty-one samples from vaccinated Australian flocks, experiencing problems with excessive Marek's disease (MD), were tested for the presence of transmissible MD viruses (MDV). Of the 16 samples which contained a transmissible agent, 14 were pathogenic in chickens, based on the development of MD lesions or depression of the bursa/body weight ratio. Of the pathogenic isolates which have been successfully typed 10 were serotype 1, and one was serotype 2 MDV. Pathogenicity of isolates varied. Several isolates caused tumours in 20-30% of both vaccinated and unvaccinated chickens. Two isolates, MPF6 and MPF23, caused tumours in more than 50% of chickens. When MPF6 and MPF23 were tested in vaccine trials bivalent vaccine gave no better protection against development of MD lesions than a monovalent vaccine. Isolate MPF23 was so pathogenic that lesions were produced in all chickens, regardless of the vaccine protocol used. Therefore vvMDV have been isolated in Australia, and unlike the vaccines tested overseas, bivalent Australian vaccines do not appear to provide greater protection against these vvMDV.     

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.     

Antibody response of chickens to serotype 1, 2, or 3 Marek's disease vaccines based on ELISA with infected cells as antigen

An enzyme-linked immunosorbent assay (ELISA) was applied to evaluate the antibody response of commercial White Leghorn chickens to vaccination against Marek's disease (MD) at hatch (day 0) with serotype-1 (Rispens), -2 (SB-1), or -3 (turkey herpesvirus, HVT) vaccine virus and to challenge on day 21 with MD virus. Antigens for the test were whole chicken embryo fibroblast cells infected with Rispens, SB-1, or HVT. The chickens were progeny of stock that had been vaccinated with HVT, and on day 21 the nonvaccinated group had higher levels of maternal antibodies to HVT than to other antigens (P < 0.05). Only SB-1 vaccine had induced antibodies by day 21, and this was detected only against homologous antigens. On day 49, all three vaccines had induced higher levels of antibodies to homologous than to heterologous antigens. Marek's Disease virus (MDV) induced antibodies to all three antigens, but challenging vaccinated chicks did not significantly increase levels of antibodies on day 81 to any of the three antigens. It was concluded that an ELISA using whole cells as antigens would have potential value for monitoring the antibody response induced by MD vaccines and virulent MDV.     

Recent increase of Marek's disease in Korea related to the virulence increase of the virus

The incidence of Marek's disease (MD), an important neoplastic disease of chickens, suddenly increased in 1997 in Korea. Most MD cases of this country were detected in chickens over 20 wk of age. Five MD viruses were isolated from field flocks in which severe MD losses had occurred, and one of the viruses was studied to compare its pathotype with the prototype JM strain. The isolate KOMD-IC induced severe depression not only in body weight but also in relative bursal weight, and the depression by KOMD-IC was more severe than that induced by JM strain. In addition, the incidence of MD tumor caused by KOMD-IC was higher than that caused by the JM strain. The protective capacity of several MD vaccines was studied against challenge with KOMD-IC. The protective levels of several MD vaccines such as herpesvirus of turkeys (HVT), HVT plus SB1, and Rispens were usually lower against challenge with KOMD-IC than those challenged with JM strain, even if the chickens vaccinated with serotype 1 were not completely protected against challenge with KOMD-IC. The above results indicate that the virulence of KOMD-IC isolated recently was increased, and the increase of MD outbreak in Korea may be related to the virulence increase of the virus. Various MD vaccine programs were applied to reduce MD loss to a broiler breeder farm where severe MD loss had occurred. Serotype 1 vaccine could dramatically decrease the mortality due to MD, and the best results were obtained from the flocks vaccinated with bivalent vaccine of Rispens and HVT.     

New serotype 2 and attenuated serotype 1 Marek's disease vaccine viruses: comparative efficacy

Attempts were made, through selection of optimum viral strains, to develop improved vaccines against Marek's disease (MD). Seven attenuated serotype 1 strains and 22 avirulent serotype 2 strains, both alone and in combination with the FC126 strain of serotype 3, were screened for protective efficacy against challenge with virulent and very virulent MD viral strains. The three viruses selected as most promising were evaluated alone and in various combinations and compared with commercially available vaccines, including FC126, bivalent (FC126 + SB-1), and CV1988/C, in 12 separate assays. Two of these new viruses--301B/1 (serotype 2) and Md11/75C/R2 (serotype 1)--were exceptionally protective compared with prototype vaccine strains. Four new monovalent and polyvalent vaccines based on these two isolates protected chickens better than FC126 alone or CV1988/C alone. Three of these new vaccines provided better protection than the bivalent (FC126 + SB-1) vaccine. Protective synergism was noted commonly between viruses of serotypes 2 and 3 but only sporadically between serotypes 1 and 2 or between serotypes 1 and 3. Strain CVI988/C was protective but was no better than FC126 alone, and it was less effective than bivalent (FC126 + SB-1) vaccine, even when used as a bivalent vaccine with FC126 or SB-1.     

Effect of diluting Marek's disease vaccines on the outcomes of Marek's disease virus infection when challenged with highly virulent Marek's disease viruses

Dilution of Marek's disease (MD) vaccines is a common practice in the field to reduce the cost associated with vaccination. In this study we have evaluated the effect of diluting MD vaccines on the protection against MD, vaccine and challenge MD virus (MDV) kinetics, and body weight when challenged with strains Md5 (very virulent MDV) and 648A (very virulent plus MDV) by contact at day of age. The following four vaccination protocols were evaluated in meat-type chickens: turkey herpesvirus (HVT) at manufacturer-recommended full dose; HVT diluted 1:10; HVT + SB-1 at the manufacturer-recommended full dose; and HVT + SB-1 diluted 1:10 for HVT and 1:5 for SB-1. Vaccine was administered at hatch subcutaneously. One-day-old chickens were placed in floor pens and housed together with ten 15-day-old chickens that had been previously inoculated with 500 PFU of either Md5 or 648A MDV strains. Chickens were individually identified with wing bands, and for each chicken samples of feather pulp and blood were collected at 1, 3, and 8 wk posthatch. Body weights were recorded at 8 wk for every chicken. Viral DNA load of wild-type MDV, SB-1, and HVT were evaluated by real time-PCR. Our results showed that dilution of MD vaccines can lead to reduced MD protection, reduced relative body weights, reduced vaccine DNA during the first 3 wk, and increased MDV DNA load. The detrimental effect of vaccine dilution was more evident in females than in males and was more evident when the challenge virus was 648A. However, lower relative body weights and higher MDV DNA load could be detected in chickens challenged with strain Md5, even in the absence of obvious differences in protection.     

Field trial in commercial broilers with a multivalent in ovo vaccine comprising a mixture of live viral vaccines against Marek's disease,infectious bursal disease,Newcastle disease,and fowl pox

A multivalent in ovo vaccine (MIV) was tested for safety and efficacy in a commercial broiler complex. The MIV comprised five replicating live viruses including serotypes 1, 2, and 3 of Marek's disease virus (MDV), an intermediate infectious bursal disease virus (IBDV) and a recombinant fowl poxvirus (FPV) vector vaccine containing HN and F genes of Newcastle disease virus (NDV). The performance of MIV-vaccinated broilers was compared with that of hatchmates that received turkey herpesvirus (HVT) alone (routinely used in ovo vaccine in the broiler complex). The chickens that hatched from the MIV-injected and HVT-injected eggs were raised under commercial conditions in six barns. Barn 1 housed 17,853 MIV-vaccinated chickens and each of the barns 2-6 housed 18,472-22,798 HVT-vaccinated chickens. The HVT-vaccinated chickens were given infectious bronchitis virus (IBV) and NDV vaccines at hatch and at 2 wk of age. The MIV-vaccinated chickens received IBV vaccine at hatch and IBV + NDV at 2 wk of age. The relative values of hatchability of eggs, livability and weight gain of chickens, and condemnation rates at processing were comparable between the MIV and the HVT groups (P > 0.05). Chickens from the MIV- and the HVT-vaccinated groups were challenged with virulent viruses under laboratory conditions. The resistance of vaccinated chickens against Marek's disease could not be assessed because of high natural resistance of unvaccinated commercial broilers to virulent MDV. The relative resistances of the MIV- and the HVT-vaccinated groups, respectively, against other virulent viruses were as follows: IBDV, 100% for both groups; NDV, 81% vs. 19%; FPV, 86% vs. 0%. The successful use of MIV under field conditions expands the usefulness of the in ovo technology for poultry.     

Embryo vaccination against Marek's disease with serotypes 1, 2 and 3 vaccines administered singly or in combination

Marek's disease virus (MDV) vaccines of serotypes 1 and 2 administered in 18-day-old embryonated eggs induced better protection against post-hatch challenge at 3 days with virulent MDV than vaccines given at hatch. Embryonal vaccination with a polyvalent vaccine containing equal quantities of serotypes 1 and 2 of MDV and serotype 3 virus (turkey herpesvirus, HVT) was also significantly more effective than post-hatch vaccination. These and earlier results indicate that protective efficacy of single or combined Marek's disease vaccine serotypes against post-hatch challenge at 3 days can be substantially improved if the vaccines are injected into 18-day embryos rather than at hatch. Injection of vaccines of serotypes 1 or 2 into embryonated eggs or hatched chicks did not cause detectable gross or microscopic lesions in chickens. Vaccine viruses of serotypes 1 and 2 could be isolated from spleen cells of chickens 1 week post-vaccination, and the titer of recoverable viruses was higher in chickens that received the vaccines at the 18th day of embryonation than in chickens vaccinated at hatch. Although embryo vaccination with HVT usually provided better protection than post-hatch vaccination against early post-hatch challenge with variant pathotypes of MDV, the protection was poor regardless of vaccination protocol. If challenge with variant pathotypes of MDV was delayed until embryonally or post-hatch HVT-vaccinated chickens were 21 days of age, protection of chickens by HVT was not enhanced. Thus, resistance induced by embryonal vaccination with HVT was qualitatively similar to that induced by post-hatch vaccination with this virus.     

鸡马立克氏病活疫苗免疫效力比较试验

用ＨＶＴ冻干苗、ＨＶＴ细胞结合苗、ＣＶＩ９８８细胞结合苗、ＳＢ１＋ＦＣ１２６双价活疫苗、３０１Ｂ／１＋ＦＣ１２６双价活疫苗和Ｚ４＋ＦＣ１２６双价活疫苗等６种鸡马立克氏病（ＭＤ）疫苗免疫ＳＰＦ白来航鸡或普通伊莎鸡，用鸡马立克氏病病毒（ＭＤＶ）强毒ＧＡ株、京－１血毒以及鸡马立克氏病超强毒ｖｖＭＤＶ－Ｍｄ５毒株分别攻击进行免疫效力比较试验。试验表明，ＭＤ单价苗的免疫效力强弱顺序依次是ＣＶＩ９８８、ＨＶＴ细胞结合苗和ＨＶＴ冻干苗，这３种ＭＤ单价苗均能给免疫鸡群提供有效的免疫保护力。ＳＢ１＋ＦＣ１２６、Ｚ４＋ＦＣ１２６和３０１Ｂ／１＋ＦＣ１２６等３种ＭＤ双价苗免疫效力显著高于ＭＤ单价苗，均能给免疫鸡群提供较强的免疫保护力，并能有效地抵抗ｖｖＭＤＶ－Ｍｄ５毒株的致瘤作用。Ｚ４＋ＦＣ１２６和３０１Ｂ／１＋ＦＣ１２６ＭＤ双价苗免疫效力无显著差异     

Protective efficacy of Marek's disease virus (MDV) CVI-988 CEF65 clone C against challenge infection with three very virulent MDV strains

Comparative 50% protective dose (PD50) assays were performed using a plaque-purified preparation of Marek's disease virus (MDV) strain CVI-988 at the 65th chicken embryo fibroblast (CEF) passage level (MDV CVI-988 CEF65 clone C) and three commercial MD vaccines: herpesvirus of turkeys (HVT) FC126, MDV CVI-988 CEF35, and a bivalent vaccine composed of HVT FC126 and MDV SB-1. In addition, comparative PD50 assays were performed in groups of chickens with maternal antibody to each of the three vaccines. Three representatives of the newly emerged biovariant very virulent (vv) MDV strains-RB/1B, Tun, and Md5-were employed as challenge virus. The experiments made feasible the differentiation between virulent MDV and vvMDV strains, within serotype 1. Vaccination with CVI-988 clone C vaccine resulted in PD50 estimates of about 5 plaque-forming units (PFUs) against challenge infection with each of the three vvMDV strains. The PD50 estimate of CVI-988 clone C vaccine was 12-fold below the PD50 of HVT FC126. The protective synergism of bivalent vaccine, composed of HVT and SB-1, was confirmed by groups given the lowest vaccine doses. The bivalent vaccine, however, resulted in incomplete protection in groups given the highest vaccine doses. Homologous maternal antibodies to serotype 1 caused a fivefold increase in the PD50 estimate of CVI-988 clone C. Heterologous maternal antibodies against HVT did not interfere with efficacy of CVI-988 clone C vaccination. However, the combination of maternal antibodies against both HVT and SB-1 (serotypes 2 and 3) showed a strong adverse effect on CVI-988 clone C vaccine.(ABSTRACT TRUNCATED AT 250 WORDS)     

鸡马立克氏病三价疫苗的安全性及免疫效力研究

将MD三价疫苗以25000PFU／只的剂量颈部皮下接种1日龄SPF雏鸡，结果表明，疫苗的接种不影响鸡体重的增加；不引起鸡法氏囊、脾脏等组织器官发生MD组织病理学变化，对鸡安全无毒性。用SPF鸡评价MD三价疫苗的免疫效力，三批疫苗对RB1B超强毒株攻击的平均保护效力为95．99％，而且无论是用RB1B超强毒株还是用BJMDV-1血毒攻击，MD三价疫苗的保护效力明显高于HVT＋SB1二价疫苗及HVT冻干苗，好于CV1988疫苗；接种MD三价疫苗的4个品系的商品鸡群，抗RB1B超强毒株攻毒的平均保护效力为94．60％；在模拟MD强毒自然传染的试验中，MD三价疫苗的保护效力达到95．65％。上述效力试验的结果说明：MD三价疫苗的免疫接种可使鸡形成抗MD强毒攻击的坚强免疫力。     

Acid alpha naphthyl acetate esterase reacting lymphocytes in the peripheral blood of chickens challenged with Marek's disease after vaccination with three different vaccines

The average percentage of acid alpha naphthyl acetate esterase reacting lymphocytes (APARL) was enumerated in the peripheral blood of chickens challenged with Marek's disease after vaccination with either turkey herpesvirus (HVT), inactivated Marek's disease virus (IMDV) or a mixture of the two (bivalent vaccine). A gradual increase in APARL value was noticed in the vaccinated chickens from day 7 to 70 after challenge with a virulent Marek's disease virus. The increase was consistent and significantly higher in bivalent (HVT plus IMDV) than in HVT-vaccinated chickens while the slight increase noticed in IMDV vaccinated-challenged birds was inconsistent.     

马立克氏病双价苗的研究Ⅲ.双价苗与单价苗免疫效应的比较病理学研究

选用SPF种蛋孵化的京白鸡60只,随机分为三组,即马立克氏病单价苗组(2000PFU HVT)、双价苗组(2000 PFU HVT 1200 PFU SB—1)以及攻毒对照组.应用组织学和透射电镜等手段对两种疫苗的免疫保护效应进行了比较病理学研究.结果证明,双价苗免疫鸡,MD发病数显著减少,发病时间推迟,病变程度明显减轻,法氏囊有大量具有分泌活性的浆细胞;单价苗免疫鸡发病数较对照组略有减少,但对延缓或减轻MD肿瘤的发生无明显作用,在法氏囊网状细胞胞浆中、核碎片附近和细胞间隙中存在大量无囊膜的马立克氏病病毒粒子,残存的细胞发生变性、坏死.由此认为,双价苗对实验性鸡马立克氏病的免疫效果明显优于HVT单价苗.     

The efficacy of recombinant fowlpox vaccine protection against Marek's disease: its dependence on chicken line and B haplotype

Earlier studies have shown that the B haplotype has a significant influence on the protective efficacy of vaccines against Marek's disease (MD) and that the level of protection varies dependent on the serotype of MD virus (MDV) used in the vaccine. To determine if the protective glycoprotein gene gB is a basis for this association, we compared recombinant fowlpox virus (rFPV) containing a single gB gene from three serotypes of MDV. The rFPV were used to vaccinate 15.B congenic lines. Nonvaccinated chickens from all three haplotypes had 84%-97% MD after challenge. The rFPV containing gB1 provides better protection than rFPV containing gB2 or gB3 in all three B genotypes. Moreover, the gB proteins were critical, since the B*21/*21 chickens had better protection than chickens with B*13/*13 or B*5/*5 using rFPV with gB1, gB2, or gB3. A newly described combined rFPV/gB1gEgIUL32 + HVT vaccine was analyzed in chickens of lines 15 x 7 (B*2/*15) and N (B*21/*21) challenged with two vv+ strains of MDV. There were line differences in protection by the vaccines and line N had better protection with the rFPV/gB1gEgIUL32 + HVT vaccines (92%-100%) following either MDV challenge, but protection was significantly lower in 15 X 7 chickens (35%) when compared with the vaccine CVI988/Rispens (94%) and 301B1 + HVT (65%). Another experiment used four lines of chickens receiving the new rFPV + HVT vaccine or CVI988/Rispens and challenge with 648A MDV. The CVI 988/Rispens generally provided better protection in lines P and 15 X 7 and in one replicate with line TK. The combined rFPV/gB1gEgIUL32 + HVT vaccines protected line N chickens (90%) better than did CVI988/Rispens (73%). These data indicate that rFPV + HVT vaccines may provide protection against MD that is equivalent to or superior to CVI988/ Rispens in some chicken strains. It is not clear whether the rFPV/gB1gEgIUL32 + HVT vaccine will offer high levels of protection to commercial strains, but this vaccine, when used in line N chickens, may be a useful model to study interactions between vaccines and chicken genotypes and may thereby improve future MD vaccines.     

不同鸡MD疫苗免疫鸡及强毒攻击后羽囊排毒规律

分别以７种鸡ＭＤ疫苗免疫ＳＰＦ鸡和狼山鸡,用琼脂扩散试验（ＡＧＰ）检查鸡群ＭＤＶ强毒攻击后不同时期的羽囊抗原,结果表明,免疫组鸡羽囊排毒高峰推迟,排毒率下降,排毒高峰维持时间短,不同疫苗免疫不同品种鸡后排毒情况有差异,ＣＶＩ９８８和两种二价苗效果优于ＨＶＴ苗。     

FIELD TRIALS WITH LYOPHILISED HERPESVIRUS OF TURKEYS VACCINE AGAINST MAREK''S DISEASE IN LAYER AND BROILER REARING FLOCKS

The safety and immunogenicity of commercial lyophilised herpesvirus of turkeys (HVT) vaccine against Marek's disease (MD) was evaluated in 19 field trials involving 76,848 vaccinated and 87,590 control chickens. In the first series of 11 trials involving one commercial vaccine at a dose of 536 plaque forming units (PFU) there was a significant reduction in the incidence of MD (P less than 0.001) and in total mortality (P less than 0.001) from 0 to 18 weeks due to vaccination and the overall protection was 85.2%. In the second series of eight trials involving another commercial vaccine at a dose range of 1360 to 1900 PFU there was a significant reduction in the incidence of MD (P less than 0.001) from 0 to 18 weeks of age due to vaccination. The overall protection was 77.8%. In both series there was a significant difference in MD mortality between meat breeders and crossbred pullets (P less than 0.005) in response to vaccination. There was also a significant difference in total mortality from 0-6 weeks of age between the two series suggesting either differences in response to the two vaccines or differences in the vaccination technique. In a number of trials the incidence of MD was low in the controls, however, where MD incidence was above 2% in the controls protection usually exceeded 80%. It was concluded the vaccines were safe and efficacious and the degree of protection was comparable to that obtained by HVT vaccines described in other countries.     

Compatibility of turkey herpesvirus-infectious bursal disease vector vaccine with Marek's disease rispens vaccine injected into day-old pullets

Mixtures of turkey herpesvirus (HVT) and Rispens poultry vaccines have been used worldwide for over 20 yr, mainly for vaccination of future layers and breeders. With increasing virulence of Marek's disease (MD) virus strains, vaccination strategies are evolving toward the use of vaccines combining HVT and Rispens. A single vaccination either in ovo or at 1 day of age with the HVT + infectious bursal disease (IBD) vector vaccine is efficient against IBD. However, with vaccination programs that include a hatchery administration of the HVT + IBD vaccine, additional protection against very virulent and very virulent-plus MD viruses is needed, especially for layers and breeders. This study looked at the combination of four commercially available Rispens vaccines with the HVT + IBD vector vaccine injected at 1 day of age. MD challenge tests that were superior to 90% in relative score in all the groups vaccinated with both vaccines showed that the mixture of HVT + IBD and Rispens vaccines had no effect on clinical protection against MD, and IBD challenge tests showed that the mixture of HVT + IBD and Rispens vaccines had no effect on clinical protection against IBD, which was equal to 100% protection in all the groups vaccinated with both vaccines.     

A comparative evaluation of the protective efficacy of rMd5deltaMeq and CVI988/ Rispens against a vv+ strain of Marek's disease virus infection in a series of recombinant congenic strains of White Leghorn chickens

Marek's disease (MD) is a lymphoproliferative disease of domestic chickens caused by a highly infectious, oncogenic alpha-herpesvirus known as Marek's disease virus (MDV). MD is presently controlled by vaccination. Current MD vaccines include attenuated serotype 1 strains (e.g., CVI988/Rispens), avirulent serotype 2 (SB-1), and serotype 3 (HVT) MDV strains. In addition, recombinant MDV strains have been developed as potential new and more efficient vaccines to sustain the success of MD control in poultry. One of the candidate recombinant MDV strains, named rMd5deltaMeq, was derived from Md5, a very virulent strain of MDV lacking the MDV oncogene Meq. Our earlier reports suggest that rMd5deltaMeq provided protection equally well or better than commonly used MD vaccines in experimental and commercial lines of chickens challenged with very virulent plus (vv+) strains of MDV. In this study, maternal antibody-positive (trial 1) and negative (trial 2) chickens from a series of relatively MD resistant lines were either vaccinated with the rMd5deltaMeq or CVI988/Rispens followed by infection of a vv+ strain of MDV, 648A, passage 10. This report presents experimental evidence that the rMd5deltaMeq protected significantly better than the CVI988/Rispens (P < 0.01) in the relatively resistant experimental lines of chickens challenged with the vv+ strain of MDV. Together with early reports, the rMd5deltaMeq appeared to provide better protection, comparing with the most efficacious commercially available vaccine, CVI988/Rispens, for control of MD in lines of chickens regardless of their genetic background.     

Comparison of blood and feather pulp samples for the diagnosis of Marek's disease and for monitoring Marek's disease vaccination by real time-PCR

Comparison of blood and feather pulp (FP) samples for the diagnosis of Marek's disease (MD) and for monitoring Marek's diseases vaccination in chickens (serotypes 2 and 3 vaccines) by real time-PCR was evaluated. For diagnosis of MD, quantification of serotype 1 Marek's disease virus (MDV) DNA load was evaluated in 21 chickens suffering from MD. For each chicken, samples of blood and FP were collected and MDV DNA load was quantified. Solid tumors are the sample of choice for MD diagnosis by real time-PCR and, hence, 14 solid tumors were included in the study as positive controls. Load of MDV DNA in FP was equivalent to that detected in solid tumors (threshold cycle [Ct] ratio above 1.7). MDV DNA load in blood samples was lower than in solid tumors and FP samples. Nonetheless, there was a statistically significant correlation of the results obtained from FP and blood (r = 0.92). Results of the Pearson correlation test showed that Ct ratio values of 1.7 in FP correspond to Ct ratio values of 1.2 in peripheral blood. For monitoring vaccines, serotypes 2 and 3 MDV DNA load was evaluated in blood and FP samples of vaccinated chickens. Serotype 2 MDV DNA load was evaluated in samples of blood and FP from 34 chickens vaccinated with SB-1 strain. Serotype 3 MDV DNA load was evaluated in blood and FP samples from 53 chickens vaccinated with HVT strain. For both serotypes, frequency of positive samples and load of vaccine DNA was higher in FP than in blood samples. There was not a statistically significant correlation between the load of SB-1 DNA (r = 0.17) or HVT DNA (r = -0.04) in FP and blood. Our results show that the load of serotypes 1, 2, and 3 DNA is higher in FP than in blood. Diagnosis of MD could be done using both FP and blood samples. Monitoring of MD vaccination by real time-PCR required the use of FP samples. There were a high percentage of false negative samples when using blood to detect serotypes 2 and 3 MDV by real time-PCR.