Egg transmission of egg drop syndrome 1976 virus in fowl

Summary

Egg transmission of egg drop syndrome 1976 virus (BC14 virus) in fowl was demonstrated in the second and third week after experimental infection. Eggs of BC14 virus infected hens were incubated weekly after disinfection with formaline gas. After 18 days of incubation, eggs with live embryos were homogenized. This egg material was fed to adult hens, housed in isolators.

Seroconversion in these birds demonstrated egg transmission.

It is suggested that egg transmission occurs as a result of viremia.     

Serological examination and egg production of progeny of fowl experimentally infected with egg drop syndrome 1976 virus

Following EDS'76 virus (BC14 virus) infection of breeder chickens by the conjunctival route, vertical transmission occurred in the first week after infection. In the progeny which had been infected with EDS'76 virus by the vertical route, increasing haemagglutination inhibiting (HI) titres to BC14 virus and increasing numbers of birds with HI titres were observed from 3 weeks to 15 weeks of age. Sixty-one per cent of the hens and 77 per cent of the cocks had 2 log HI BC14 virus titres exceeding 4 at an age of 15 weeks. Some birds which han been serologically negative throughout the rearing period, seroconverted between 25 and 28 weeks of age. This phenomenon occurred in hens as well as in cocks. Simulation of stress twice during the laying period by injection of corticosteroid hormone did not increase the number of birds serologically positive to EDS'76 virus. EDS'76 was observed in the group of hens that was vertically infected, since egg production was significantly depressed between 28 and 34 weeks of age. Probably this was mainly the results of a production drop in the hens showing serconversion at 27 or 28 weeks of age. In this group of fowl vertically infected with EDs'76 virus, serologically positive birds appeared to be protected for the greater part to BC14 virus challenge at 50 weeks of age, while negative birds seemed to be fully susceptible. Chicks hatched from eggs collected in the third and fourth week after infection of the dams had maternal antibodies. Fertility and hatchability of apparently normally shelled eggs seemed not to be affected after BC14 virus infection of the dams. Intensive contact with contaminated faeces is probably an indispensable condition for lateral transmission of the virus.     

Serological examination and egg production of progeny of fowl experimentally infected with Egg Drop Syndrome 1976 virus

Summary

Following EDS'76 virus (BC14 virus) infection of breeder chickens by the conjunctival route, vertical transmission occurred in the first week after infection. In the progeny which had been infected with EDS'76 virus by the vertical route, increasing haemagglutination inhibiting (HI) litres to BC14 virus and increasing numbers of birds with HI litres were observed from 3 weeks to 15 weeks of age. Sixty‐one per cent of the hens and 77 per cent of the cocks had ²log HI BC14 virus litres exceeding 4 at an age of 15 weeks.

Some birds which had been serologically negative throughout the rearing period, seroconverted between 25 and 28 weeks of age. This phenomenon occurred in hens as well as in cocks. Simulation of stress twice during the laying period by injection of corticosteroid hormone did not increase the number of birds serologically positive to EDS'76 virus.

EDS'76 was observed in the group of hens that was vertically infected, since egg production was significantly depressed between 28 and 34 weeks of age. Probably this was mainly the result of a production drop in the hens showing seroconversion at 27 or 28 weeks of age.

In the group of fowl vertically infected with EDS'76 virus, serologically positive birds appeared to be protected for the greater part to BC14 virus challenge at 50 weeks of age, while negative birds seemed to be fully susceptible. Chicks hatched from eggs collected in the third and fourth week after infection of the dams had maternal antibodies. Fertility and hatchability of apparently normally shelled eggs seemed not to be affected after BC14 virus infection of the dams. Intensive contact with contaminated faeces is probably an indispensable condition for lateral transmission of the virus.     

Experimental infection of laying chickens with egg-drop syndrome 1976 virus

Brown layer hens (BC and HC strains) and white layer hens (WL strain) orally infected with the H-162 strain of the egg-drop syndrome 1976 virus developed few clinical signs except for abnormal egg production. Depressed and/or aberrant-egg production was observed for 3 days or longer in 17 of 18 BC hens, 13 of 15 HC hens, and 10 of 17 WL hens. On the average, abnormal egg production began 8.8, 10.3, and 12.2 days after infection of the BC, HC, and WL hens, respectively. Egg production was depressed in the WL hens, but little depression was observed in the BC and HC hens. Aberrant-egg production was much less frequent in the WL hens than in the BC and HC hens. Aberrant eggs were shell-less, soft-shelled, thin-shelled, and/or discolored. No eggs of abnormal internal quality or shape were observed. The virus spread from infected BC and WL hens to contact hens.     

Effects of experimental infection of fowl with EDS'76 virus,infectious bronchitis virus and/or fowl adenovirus on laying performance

Following BCI4 (EDS'76) virus infection of brown layer hens at 33 weeks of age, production of normally shelled eggs dropped from 87 per cent to 49 per cent within 3 weeks. The production of soft shelled and shell‐less eggs attained a maximum of 33 per cent 3 weeks after infection (p.i.). Shell quality recovered completely within 5 weeks p.i.

Egg production problems in White Leghorns infected with BCI4 virus were less severe and of shorter duration than in brown layers.

Both in brown layers and in White Leghorns total egg production, mean weight of normally shelled eggs, and internal egg quality were not affected following BCI 4 virus infection at 33 and 28 weeks of age, respectively. Besides shell abnormalities no clinical disease symptoms were observed. Vaccination with a commercial EDS'76 vaccine (Nobivac EDS'76^®) at 17 weeks of age had no adverse effects on laying performance and provoked adequate immunity against challenge at 33 weeks of age. The same observations were made following BCI4 virus infection at 17 weeks of age.

After infectious bronchitis virus (IBV) (H52 virus) infection of laying fowl the percentage of eggs with shell aberrations (rough, misshapen and/ or soft shells) increased to a maximum of 8 per cent, total egg production was depressed, mean egg weight was reduced I to 2 grams, and up to 10 Per cent of normally shelled eggs showed watery, not ropy thin albumen. This abnormally thin albumen was observed in a considerably higher proportion of eggs with shell defects than in normally shelled eggs. No turbidity of the thick albumen was observed and no symptoms of respiratory disease were noticed.

The severity and duration of adverse effects of IBV infection on laying performance depend very much on the stage of production in which the infection occurs. Following IBV infection at the onset of production a much severer drop in total production and in production of normally shelled eggs, a greater increase in the number of abnormally shelled eggs, and more lasting adverse effects on egg weight and internal egg quality were observed, in comparison with infection after peak production. Compared with single infections, a combined BC14 virus and IBV infection of brown layers at 33 weeks of age resulted in greatly potentiated adverse effects on total egg production, number of eggs with aberrant internal quality, and duration of production problems.

Following a combined BC14 virus and IBV infection, in a great proportion of eggs with shell defects and watery thin albumen, turbidity of the thick albumen was observed also, probably due to combined effects on the uterus of both IBV and BC14 virus.

BC14 virus infection did not reinforce the adverse influence of IBV infection on egg weight.

The same observations as described for the combined BC14 virus and IBV infection were made following BC14 virus infection of fowl previously infected with IBV.

It is concluded that changes of internal egg quality in field cases of EDS'76 are most likely due to subclinical IBV infections.

After infection of brown layers at 33 weeks of age with fowl adenovirus 66 (FA V 66) neither symptoms of clinical disease were observed nor effects on egg production, egg weight, and egg quality. Also, in a combined infection with FA V66, IBV, and BCI4 virus, no pathogenic significance could be attributed to the FAV.     

Mycoplasma gallisepticum vaccination: further studies on egg transmission and egg production

Leghorn hens vaccinated twice with an inactivated Mycoplasma gallisepticum (MG) bacterin before egg production and subsequently challenged with virulent MG were protected against transmission of MG through the egg. Unvaccinated control hens transmitted MG through the egg at a high rate. When unvaccinated hens were vaccinated with MG bacterin 2 weeks after challenge with MG, there was no significant decrease in egg transmission. Hens vaccinated twice before laying did not suffer as severe egg-production drops as unvaccinated hens did when challenged with virulent MG. In a natural MG outbreak in a leghorn breeder flock, 50 hens were separated from the remainder of the flock and vaccinated with MG bacterin approximately 3 weeks after initial exposure. The unvaccinated hens transmitted MG through the egg at a rate three times higher than the rate of transmission of the vaccinated hens.     

Sporadic congenital transmission of avian leukosis virus in hens discharging the virus into the oviducts.

The efficacy of the albumen test for infectious avian leukosis virus (ALV) was examined in detecting congenitally transmitting hens. Seventy-three White Leghorn non-viremic hens with antibody to ALV were used. Eleven of the hens shed infectious ALV into their egg albumen, whereas only 7 of the 11 ALV-positive hens shed ALV antigens. The egg albumen test for infectious ALV was shown to be more effective in detecting the congenitally transmitting hens than that for ALV antigens. Then, twenty of the 62 hens which shed no infectious ALV into the albumen were studied for transmission of ALV to their embryos and for discharging ALV into the oviduct and vagina. Six of the 50 embryos from 4 hens were found to be infected with ALV but all of the 227 embryos from remaining 16 hens were free from the infection. Discharge of the virus into the oviduct and vagina was found both in the 4 transmitting hens and in 6 of the 16 non-transmitting hens. These results suggest that the hens discharging ALV into the oviduct, even though they do not shed ALV into egg albumen, may transmit the virus sporadically to their embryos.     

Outbreaks of egg-drop syndrome-1976 in Japan and its etiological agent

A condition similar to egg-drop syndrome-1976 (EDS-76) occurred in 14 broiler breeding flocks in 2 farms in Japan from December 1978 to January 1980, and it was diagnosed as EDS-76 by serologic and virological investigations. Egg production fell suddenly when the hens were 30 to 55 weeks of age, and the depression lasted 3 to 7 weeks. Production fell 6 to 25%. Depressed egg production was accompanied by the laying of shell-less, soft-shelled, and thin-shelled eggs associated with loss of egg-shell pigment. Eleven isolates of hemagglutinating adenovirus were isolated from cloacal swabs (10 isolates) and a uterus (1 isolate) of hens in one farm. One isolate, cloned and named JPA-1, had the same antigenicity in serologic tests and the same biological and physicochemical properties as the BC14 strain of EDS-76 virus.     

Influenza in commercial broiler breeders.

Influenza was detected in a flock of broiler breeders during routine serological monitoring. Although there were no clinical signs, egg production may have been affected in hens on one story of a two-story breeder house. Intensive measures were taken to avoid transmission to other farms. Two months after the flock was found to be serologically positive, sentinel hens were placed in the flock, and they became serologically positive 1 month later. In spite of this evidence for virus being present in the flock, no detectable transmission to any other farm occurred.     

犬瘟热病毒免疫蛋鸡血清抗体与卵黄抗体消长规律的研究

试验旨在了解产蛋鸡对犬瘟热病毒(canine distemper virus,CDV)抗原的免疫反应及其血清抗体和卵黄抗体的消长规律,为制备卵黄抗体提供依据。将CDV接种Vero细胞和DF1细胞进行传代并测定其病毒滴度,选取细胞病变出现早、病毒滴度高的Vero细胞毒作为接种病毒抗原免疫蛋鸡。每10 d免疫蛋鸡1次,第4次免疫后每30 d加强免疫1次。免疫前及免疫后每10 d采血分离血清,每5 d收集鸡蛋提取卵黄抗体,用琼脂扩散法和间接ELISA法测定其抗体水平。结果显示,血清抗体比卵黄抗体的滴度高,两者呈正相关性,卵黄抗体出现较血清抗体迟3~5 d,卵黄抗体在第3次免疫后3~5 d就已达到较高水平,此时即可开始收集鸡蛋制备卵黄抗体。     

Congenital transmission of avian leukosis virus in the absence of detectable shedding of group specific antigen

Congenital transmission of avian leukosis virus (ALV) in the absence of detectable amounts of group specific (gs) antigen in egg albumen was found to occur in one commercial and one specific pathogen-free (SPF) flock. The prevalence of congenitally transmitting hens which did not excrete gs antigen was particularly high in a commercial flock where 26/27 hens transmitted ALV. Some of the ALV-transmitting hens in the commercial flock had virus in vaginal swabs thus enabling infection to be detected. The reasons for such a high proportion of congenitally transmitting hens which did not shed detectable amounts of gs antigen in the commercial flock was not determined. In the SPF flock, 2/15 hens congenitally transmitted ALV although virus could not be detected in vaginal swabs, whole blood or egg albumen and antibodies to subgroups A or B were not present. This form of ALV-infection persisted in two successive generations. These results indicate the necessity of testing for infectious ALV in embryos, in order to ascertain that a flock is genuinely free of ALV.     

Egg transmission of avian adenovirus-associated virus and CELO virus during a naturally occurring infection.

Both avian adenovirus-associated virus (A-AV) and CELO virus were isolated from the embryonating eggs of 25-week-old black sex-linked hens during a naturally occurring infection. In the first 7 days of egg collection, A-AV was isolated from 10 of 43 (23.2%) embryonating eggs, and CELO virus was isolated from 8 of 43 (18.6%) embryonating eggs. Both viruses were isolated from six eggs. In the next 16 days of egg collection, A-AV and CELO virus were coisolated from 1 of 127 (0.8%) eggs; all other samples were negative for both viruses. All six hens transmitting A-AV to eggs and 5 of 6 hens transmitting CELO virus showed seroconversions (fourfold increase in antibody concentrations). Viruses were not isolated from eggs after the hens showed a fourfold increase in antibody concentrations.     

蛋鸡首免乙型脑炎病毒后卵黄抗体变化规律的研究

本试验旨在研究疫苗、免疫剂量和注射方式对卵黄抗体效价规律的影响,探讨制备抗猪乙型脑炎病毒卵黄抗体蛋鸡的最佳免疫程序。选用无免疫褐羽蛋鸡180只,随机分成18组,每组10只。1、2组均为对照组,注射无菌生理盐水;3~10组采用皮下和肌肉两种注射方式,并依次注射灭活苗0.2、0.5、1.0和1.5 mL;11~18组同样采取两种注射方式,依次免疫相同剂量弱毒苗。各试验组分别于免疫前1 d、免疫后第3、7、10、14、18、21和28天采集当日鸡蛋6枚,提取卵黄抗体,测定效价。试验结果显示,1~6、11、12组卵黄抗体效价均为0,未产生明显免疫应答;7~10组注射灭活苗后7 d,平均卵黄抗体效价达到峰值,抗体持续时间为14 d;13~18组注射弱毒苗后14 d达到最高值,抗体持续时间为21 d;注射剂量相同但注射方式不同的两组之间比较,卵黄抗体效价差异不显著（P>0.05）;相同注射方式,相同疫苗的各试验组间,随着免疫剂量的增加卵黄抗体效价逐渐加强,且差异显著（P<0.05）。以上结果表明,肌肉或皮下两种注射方式,蛋鸡产生明显免疫应答至少需要免疫灭活苗1.0 mL或弱毒苗0.5 mL。比较弱毒苗与灭活苗,灭活苗刺激机体产生的抗体速度较快,但维持时间较短;较少剂量弱毒苗就可刺激机体产生抗体,但速度慢、维持时间长。     

CDV免疫蛋鸡血清抗体与卵黄抗体的消长规律及其相关性分析

为了解产蛋鸡对犬瘟热病毒杭原的免疫反应及其血清抗体和卵黄抗体的消长规律及相关性,为制备卵黄抗体提供依据,将犬瘟热病毒接种Vero细胞和DF1细胞进行传代并测定其病毒滴度,选取细胞病变出现早,病毒滴度高的Vero细胞毒作为接种病毒抗原免疫蛋鸡。通过每10d进行蛋鸡免疫一次,四免后每30d加强免疫一次的方法进行免疫。免疫前和免疫后每10d采血分离血清和每5d收集鸡蛋提取卵黄抗体,用琼脂扩散法和间接ELISA法测定其抗体水平。结果显示,血清抗体比卵黄抗体的滴度高,两者间呈正相关性,卵黄抗体出现较血清抗体迟3～5d,卵黄抗体在三免后3～5d就已达到较高水平,此时即可开始收集鸡蛋制备卵黄抗体。     

Study on the Change Rule of Egg Yolk Antibody in Laying Hens after First Immunization by Japanese Encephalitis Virus

The purpose of this experiment was to study the immunization rule of the egg yolk antibody affected by different vaccines,immunization dose and injection ways and further to discuss the optimal immunization procedures of the laying hens for the preparation of egg yolk antibody against swine Japanese encephalitis virus.180 brown laying hens without any vaccines were selected and divided into 18 groups randomly,each group of 10 hens.Groups 1,2 were the control groups,injected with the sterile saline;Groups 3 to 10 were injected with subcutaneous or intramuscular injection,and the vaccine was injected with 0.2,0.5,1.0 and 1.5 mL successively.Groups 11 to 18 were also adopted two kinds of injection,followed by the same dose of vaccine immunization.Six eggs of each experimental group were gathered before immune day and after 3,7,10,14,18,21 and 28 days,the egg yolk antibody was extracted and the titer was determined.As a result,the egg yolk antibody titers of groups 1 to 6,11 and 12 were all 0,and no significant immune response produced;The hens from 7 to 10 groups were injected with the inactivated vaccine.After 7 days,the average antibody titer reached the peak,and the duration of the antibody was 14 days.The hens from 13 to 18 groups were injected with the attenuated virus vaccine.After 14 days,the average antibody titer reached the highest value,and the duration of the antibody was 21 days.The egg yolk antibody titers were not significantly different in the two compared experiment groups with the same injection dose but with different injection ways (P>0.05).With the same injection way of each experiment group,and the difference was significant (P>0.05).Compared with some groups with the same injection and vaccine,the titer of yolk antibody was gradually increased with the increase of the immune dose,and the difference was significant (P<0.05).The results showed that,no matter intramuscular or subcutaneous injection,in order to produce a significant immune response to hens,the immune antigen dose was 1.0 mL inactivated vaccine or 0.5 mL attenuated vaccine at least.Compared with the attenuated and inactivated vaccine,inactivated vaccine stimulated the body to produce the antibody faster,but the maintenance time was shorter;The lower dose of attenuated vaccine could stimulate the body to produce antibodies,but the speed was slower,the maintenance time was longer.     

Seroepidemiology of marble spleen disease on a large pheasant farm in california

An agar gel precipitation (AGP) test was used to study the epidemiology of marble spleen disease (MSD) on a large pen-raised pheasant farm with a prior history of MSD. Tests conducted during early season egg production (March 1983) on previously infected breeders, poults from these breeders, and MSD-free hens introduced onto the farm, indicated absence of MSD antibodies (and clinical disease) during the breeding season (February–July 1983). Thus, a carrier state or egg transmission could not be demonstrated. The disease did not occur in late summer (September) in the same area of the ranch where it was diagnosed for the past 5 seasons. The AGP test was useful in confirming the pattern of the disease spread which moved erratically but within the general area of the index pen. Antibodies were detected in penmates approximately 10–15 days after the first observed mortality, but did not persist after the disease had run its course. Our observations suggest that the reservoir of the virus exists in the environment, and mechanical transmission (caretaker, equipment etc.) is important in sustaining the disease during an outbreak.     

Efficacy of infectious bronchitis virus vaccines against heterologous challenge

Twenty-four-week-old white Leghorn layers were inoculated subcutaneously with a killed Newcastle disease-infectious bronchitis (Massachusetts type) virus (MIBV) vaccine. Twenty-eight weeks after vaccination, the birds were challenged intraocularly with the Arkansas strain of infectious bronchitis virus (AIBV) to determine the effects of heterologous virus exposure on egg production, egg quality and serum antibody response of the birds. The challenged hens laid significantly (P less than 0.005) fewer eggs than the unchallenged layers. Eggs laid by the unchallenged groups weighed significantly more (P less than 0.005) than those laid by the challenged groups. Further, the internal quality (Haugh units) and shell quality of eggs laid by the AIBV-challenged hens was significantly (P less than 0.005) inferior to those from the unchallenged hens. In addition, the AIBV-challenged hens laid more soft-shell, misshapen and small eggs than the unchallenged hens. The Arkansas serum haemagglutination inhibition (AIBV-HI) titres of AIBV challenged birds increased up to four weeks after challenge. The corresponding MIBV haemagglutination-inhibition (MIBV-HI) titres decreased during the same period. The study indicates that killed MIBV vaccine offered no protection to birds exposed to heterologous AIBV.     

Pathogenicity and distribution of egg-drop syndrome-1976 virus (JPA-1) in inoculated laying hens

Rhode Island Red laying hens that lacked hemagglutination-inhibition antibody were inoculated orally with the JPA-1 strain of adenovirus isolated from a flock affected with egg-drop syndrome-1976 in Japan and observed for 80 days. Inoculated hens laid abnormal eggs, such as shell-less, soft-shelled, and cracked eggs and those with loss of pigmentation, from 8 days postinoculation (PI). Fifteen out of 16 hens laid abnormal eggs. Egg-production rte fell from 94% to 50% between 13 and 16 days PI. When the abnormal eggs were excluded, egg production was 17%; then it recoverd slowly, reaching 67% by the end of the experiment. The virus was recovered from various organs of inoculated hens from 3 to 7 days PI. Fluorescent antigens of the virus were observed in the epithelial cells and desquamated cells from the epithelium of the uterus and isthmus 10 and 14 days PI.     

Immune responses of breeding chickens to trivalent oil emulsion vaccines: responses to infectious bronchitis

Three similar flocks of broiler breeder parent chickens that had been given live infections bronchitis (IB) vaccines during rearing were injected at 20 weeks of age with three different oil emulsion vaccines: a commercial monovalent Newcastle disease (ND) vaccine (flock A); an experimental bivalent vaccine containing ND and infectious bursal disease (IBD) components (flock B); and an experimental trivalent vaccine containing ND, IBD and IB components (flock C). One week after vaccination 40 hens from flock A and 40 from flock C were taken to the laboratory and their egg yields individually recorded. At 37 weeks of age they were challenged by aerosol exposure to virulent IB virus. The egg production dropped significantly in the hens from flock A but not in the hens from flock C. On the farm, flock C showed a higher mean IB virus antibody titre four weeks after vaccination but titres rose in all three flocks indicating the presence of active IB virus infection. No differences in egg yields were found between the three farm flocks.     

The relationship between the magnitude of the specific antibody response to experimental salmonella enteritidis infection in laying hens and their production of contaminated eggs.

Detecting infected laying flocks is a vital part of many efforts to control egg-associated transmission of Salmonella enteritidis to humans. The relationship between the development of a specific antibody response in infected hens and the deposition of S. enteritidis in eggs is important for establishing the epidemiologic relevance of serologic testing methods. In two trials, laying hens were infected with large oral doses of phage types 13a and 14b isolates of S. enteritidis. Approximately 38% of all infected hens produced at least one contaminated egg, at an overall incidence of 5.2%, between 3 and 23 days postinoculation. As determined by enzyme-linked immunosorbent assay with an S. enteritidis flagellar antigen, 91.7% of inoculated hens produced specific serum antibodies. Although hens with very high antibody titers were associated with a significantly elevated frequency of egg contamination, a consistently direct relationship was not evident between the magnitude of the antibody responses of individual hens and the frequency at which they laid contaminated eggs. Accordingly, although serologic tests can be valuable screening tools for preliminary detection of S. enteritidis infections in poultry, the magnitude of the antibody responses detected in individual hens may not predict the overall risk of egg contamination associated with particular laying flocks.