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.     

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.     

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.     

Biochemical changes in blood and uterine fluid of fowl following experimental EDS'76 virus infection

The pH, pCO2, and pO2 values and the concentrations of sodium, potassium, calcium, magnesium, chloride, phosphorus, bicarbonate, base excess, protein, glucose, as well as the activity of alkaline phosphatase and malate dehydrogenase were determined in the venous blood and uterine fluid of control and EDS'76 virus-infected fowl. Moreover, the pH of the mucosa of different parts of the oviduct was measured. Hens were examined in the period from 10 to 24 days following infection; blood and uterine fluid samples were collected approximately 14 hours after oviposition, provided a plumped egg was present in the uterus. Examination of blood and pH measurement of oviduct mucosa did not yield significant differences between infected and noninfected hens. In comparison with noninfected control birds, the mean sodium concentration of the uterine fluid of infected hens producing soft shelled or shell-less eggs had evidently increased, while the mean concentration of potassium, calcium, magnesium and glucose had decreased. Similar differences were also observed between infected hens producing normally shelled eggs and infected hens producing abnormally shelled eggs. No significant differences between infected and not infected hens were observed concerning the other values determined in the uterine fluid. It is concluded that functional disturbances which account for shell aberrations following EDS'76 virus infection are located in the surface epithelial cells of the uterine mucosa. These disturbances are very probably initiated by a depressed function of the sodium pump. All changes observed in the uterine fluid of infected hens could be explained by this depressed function.     

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.     

Seroprevalence of egg drop syndrome - 76 virus as cause of poor egg productivity of poultry in Nsukka,South East Nigeria

To determine if egg drop syndrome 76 virus infection is among the causes of lowered egg productivity in commercial poultry farms in South Eastern Part of Nigeria and to know the prevalence of the infection, ten farms with history of lowered egg production in Nsukka local government area of Enugu State were randomly selected. Sera from ten hens in each of the selected farms were assayed for antibodies against EDS 76 virus by the haemagglutination-inhibition (HI) test. The mean HI titre of the ten hens in each of the farms was recorded as EDS - 76 antibody titre for the farm. Nine out of the 10 farms tested were positive for EDS - 76 antibodies with HI titres ranging between 16 and 256. Out of 10 flocks with production of 65% and above 9 were EDS-76 HI negative.     

Effects of infectious bronchitis virus (Arkansas strain) on laying chickens

Seventy-seven-week-old white leghorn layers were inoculated intraocularly with the Arkansas strain of infectious bronchitis virus (AIBV) to study the effects of the virus on egg production and on antibody response of the birds. Infected hens laid fewer eggs than the controls, and those eggs weighed less than eggs laid by controls. Further, the shell quality and internal quality of eggs laid by infected birds were inferior. The serum hemagglutination-inhibition (HI) titers of infected birds increased continuously through 4 weeks postinfection; serum HI titers of the controls were negligible.     

Involvement of spleen components of mature fowl in the primary and secondary humoral antibody response following experimental EDS'76 virus infection

Cellular changes in spleens of mature fowl in relation to both the primary and secondary humoral antibody response following experimental EDS'76 virus infection were studied. The influence of splenectomy on humoral antibody response was also examined. Experimental fowl had been naturally infected with fowl adenovirus (FAV) but did not possess precipitins to these viruses at the time of EDS'76 virus infection. Since EDS'76 infection provokes a recall of the group antibody to FAV, this infection simultaneously induces a primary response against EDS'76 virus and a secondary response due to the recall of the group antibody to FAV. HI and precipitating antibody to EDS'76 virus (primary response) were first detected at 6 and 8 days p.i. respectively. Curves of HI, precipitating and neutralising antibody titres were biphasic; the first peak (IgM peak) occurred at 10-11 days p.i., the second (IgG peak) at 16-28 days p.i. Precipitating antibodies to FAV (secondary response) were demonstrated from 4 days p.i. The curve of these antibody titres was also biphasic, with peaks at the same times as in the primary response. Based on HI and AGP testing of primary and secondary immune response in both splenectomised and non-splenectomised fowl it is concluded that in the primary response the spleen of the adult fowl is involved significantly in only IgM secretion, while in the secondary response it is likely that both IgM and IgG are secreted in considerable amounts. Clusters of lymphoblasts and plasmablasts were observed at 3 days p.i. in the red pulp. It is very likely that antigen-antibody complexes are formed from that time and circulate bound to the surface of lymphocytes. These antigen-loaded lymphocytes are 'picked up' from the blood stream by -red pulp macrophages, leading to enhanced formation of lymphoblasts in the red pulp. Great numbers of these cells (which are very probably IgM secreting cells) were present on days 6 and 7 p.i., but were no longer detectable after day 10 p.i. -macrophages of the macrophagal ellipsoidal corona (MEC), leading to significant enlargement of the periellipsoidal lymphoid tissue (PELT) by an increase of the number of lymphocytes observed from days 4-12 p.i. The MEC was significantly enlarged from 7-12 days p.i., very likely due to an increased number of macrophages. Following deposition of antigen in the white pulp, formation of follicles begins. The number of small, intact follicles including follicle precursors increased from 6 days p.i.(ABSTRACT TRUNCATED AT 400 WORDS)     

Resistance of chickens to challenge with the virulent Herts 33 strain of Newcastle disease virus induced by prior infection with serologically distinct avian paramyxoviruses.

Results indicate that some degree of protection from challenge by Newcastle disease virus (NDV)/Herts 33 was conferred on chickens by prior infection with PMV/turkey/Wisconsin/68, PMV/turkey/Ontario/6661/68, PMV/Netherlands/449/75 and PMV/parakeet/England/39/78 viruses, all of which are serologically related but distinguishable from NDV. Except for one bird which survived challenge three weeks after infection with Robin/Hiddensee/19/75, no protection was seen in chickens infected with other unrelated avian paramyxoviruses. In contrast to infection with NDV-B1, birds protected by infection with avian paramyxoviruses showed large increases in NDV haemagglutination inhibition (HI) titres after challenge. In these birds considerable increases in the homologous HI titres were also seen after challenge.     

Comparison of serological tests for the measurement of the primary immune response to avian infectious bronchitis virus vaccines

The immune response of individual chickens exposed in an aerosol apparatus to live commercial avian infectious bronchitis virus (IBV) vaccines was measured by serum neutralization (SN), haemagglutination inhibition (HI), complement fixation (CF) and agar gel precipitin (AGP) tests over a period of 14 weeks.The SN titres showed considerable variation for individual chickens and in a large number of birds were negative until 14 weeks after infection.Positive HI titres were recorded for most birds at one week after infection and persisted throughout the observation period. Some relationship was seen between HI and SN titres particularly in birds showing a high immune response.The results obtained with the AGP were transient, variable and did not compare well with results obtained by the other tests. The highest number of positive AGP reactors were seen two to three weeks after infection.Most birds showed positive titres by the CF test at some time after infection but titres were always low and did not correlate with results obtained by the other tests.Twenty-two weeks after vaccination 23 chickens were challenged by aerosol exposure to the Massachusetts 41 strain of IBV and four days later the trachea, kidneys and oviducts were removed from each bird for attempted virus isolation. Virus was recovered from only one kidney and 11 trachea samples. The mean pre-challenge HI and SN titres of birds from which no virus was recovered were significantly higher than the mash titres of vaccinated birds from which virus was isolated after challenge.     

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.     

OBSERVATIONS ON THE EPIDEMIOLOGY OF PORCINE PARVOVIRUS

Evidence presented suggests that porcine parvovirus is highly stable and infective. Introduction of virus to susceptible herds results in 100% infection rate within the following 3 months. Active immunity is associated with high persistent levels of haemagglutination-inhibitating (HI) antibody (> 256), piglets suckling immune sows acquiring HI titres between 10,000 and 40,000. Loss of passive immunity, measured by HI, occurs in a majority of pigs between 14 and 26 weeks of age (mean 21 weeks), whilst an average of 25% (2–47%) of pigs lose HI titres between 26 and 36 weeks of age. Susceptibility to challenge with virus does not occur until 3–5 weeks following loss of HI titres. In endemically infected herds 98–100% of adult pigs show serological evidence of active immunity.
A significant proportion of gilts may not be actively immune to porcine parvovirus at the time of first service, and subsequent infection may occur while these gilts are pregnant.     

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 Eimeria mitis on egg production of single-comb White Leghorn hens.

Single-comb white leghorn hens infected with either B4 or C2 strain of Eimeria mitis produced watery droppings as early as day 5 PI. E. mitis infection did not affect egg weight. However, specific gravities of the eggs produced by the hens infected with E. mitis were lower than those produced by the controls. Egg production was significantly reduced temporarily, but most birds returned to production within 14 days. Many of the birds that ceased to lay went through a complete-body molt (56% of the C2-infected hens and 20% of the B4-infected hens). Hens that ceased to lay regained considerable amounts of pigment in their skin, beaks, and shanks.     

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.     

Indirect method for prediction of hemagglutination inhibition antibody titers to Newcastle disease virus in chickens by titration of antibodies in egg yolk.

Attempts were made to establish methods for indirect prediction of hemagglutination inhibition (HI) antibody titers to Newcastle disease virus (NDV) in sera of laying hens and day-old chicks by determining if these are correlated to HI titers in egg yolks. For this purpose, geometric means of HI antibody titers in sera from 60 hens, yolks from 60 matched eggs, and sera from 180 day-old chicks of an identical vaccination program were measured and plotted. There was a significant correlation between HI antibody titers in yolks (X) and hens (Y), with a linear regression of Y = 23.24 + 0.47X and a correlation coefficient of r = 0.65. The linear regression between HI antibody titers in yolks (X) and chicks (Y) was Y = 6.33 + 0.36X (r = 0.58). Immunity to NDV in hens and their offspring can be maintained effectively, and the proper time for the vaccination or booster can be determined by reference to HI titers predicted from the linear regression in the present study. The approach of testing egg yolk for HI titers provides a feasible alternative to determining HI titers from blood samples and eliminates stress in birds during blood sampling.     

Review of induced molting by feed removal and contamination of eggs with Salmonella enterica serovar Enteritidis

As laying hens age, egg production and quality decreases. Egg producers can impose an induced molt on older hens that results in increased egg productivity and decreased hen mortality compared with non-molted hens of the same age. This review discusses the effect of induced molting by feed removal on immune parameters, Salmonella enterica serovar Enteritidis (SE) invasion and subsequent production of SE-contaminated eggs. Experimental oral infections with SE show molted hens are more susceptible to SE infection and produce more SE-contaminated eggs in the first few weeks post-molt compared with pre-molt egg production. In addition, it appears that molted hens are more likely to disseminate SE into their environment. Molted hens are more susceptible to SE infection by contact exposure to experimentally infected hens; thus, transmission of SE among molted hens could be more rapid than non-molted birds. Histological examination of the gastrointestinal tracts of molted SE-infected hens revealed more frequent and severe intestinal mucosal lesions compared with non-molted SE-infected hens. These data suggest that induced molting by feed deprivation alters the normal asymptomatic host-pathogen relationship. Published data suggest the highest proportion of SE-positive eggs is produced within 1-5 weeks post-molt and decreases sharply by 6-10 weeks and dissipates to the background level for non-molted hens by 11-20 weeks. Appropriate treatment measures of eggs produced in the fist 5 weeks post-molting may decrease the risk of foodborne infections to humans.     

Protection of laying hens against Salmonella Enteritidis by immunization with type 1 fimbriae

Eighteen chickens were immunized subcutaneously with purified type 1 fimbriae from Salmonella enterica serotype Enteritidis at 18 and 21 weeks of age. Evidence of IgG and IgA responses was found in the eggs and in the sera of the immunized hens. Three weeks later, immunized and non-immunized chickens (n=18) were challenged intravenously with 2x10(7) live Salmonella enterica serotype Enteritidis. There was no significant difference in the numbers of eggs laid by immunized and non-immunized birds. The percentage of Salmonella contaminated eggs was significantly higher in the non-immunized group than in the immunized group due to a higher percentage of contamination of the externally disinfected egg shells. There were no statistical differences in the percentages of contaminated yolks and egg whites between control and immunized birds. No differences in the number of colonizing bacteria could be found in the spleen nor in the liver between the immunized and the control groups throughout the experiment. Salmonella was cleared from the ovary of the immunized birds in the second week p.i., in contrast to the control birds where Salmonella was isolated till the third week after infection. Oviducts were significantly more infected in the control group than in the immunized group. Salmonella was cleared from the oviducts at 3 weeks p.i. in the immunized hens but not in the control hens. In conclusion, we demonstrated that the immunization of laying hens with type 1 fimbriae reduced the number of contaminated eggs and reduced the colonization of the reproductive organs.     

Comparison of duration of immunity in chickens infected with a live infectious bronchitis vaccine by three different routes.

Three groups of chicks were vaccinated by aerosol, intra-ocular and drinking water routes with a live infectious bronchitis (IB) vaccine. At one, two, six, 15 and 32 weeks after vaccination five birds from each group were sampled for testing for IB haemagglutination-inhibiting (HI) antibodies and challenged. Assessment of susceptibility to infection was measured by recovery of virus from individual tracheas and from kidney and gonad pools four days after challenge. Virus was isolated from all kidney and gonad pools of birds challenged one week after vaccination, the kidney and gonad pools of the drinking water vaccinates at two weeks, the kidney pool of the intra-ocular group at 15 weeks and all organ pools except the gonads of the intra-ocular group at 32 weeks. Tracheal resistance was found in most of the birds challenged one week after vaccination and in all the birds tested at two weeks but had begun to wane by six weeks after vaccination. No correlation was found between low HI antibody titres of individual birds and their susceptibility to challenge measured by reisolation of virus from the traches, but birds with titres over log2 6 were always resistant.     

Serosurvey of five viruses in chickens on smallholdings in Bangladesh

A serologic survey was undertaken in chickens in smallholdings in Bangladesh for avian influenza A virus (AIV), egg drop syndrome '76 virus (EDS'76V), infectious bronchitis virus (IBV), Newcastle disease virus (NDV) and reovirus (RV) in three phases: January 2002-May 2003, September 2003-August 2004, and August 2005-March 2006. Four hundred thirty-six sera collected in the 2nd phase, 295 in the first phase, 755 in the 1st plus 2nd phases and 295 in the 1st phase were investigated for AIV, EDS'76V, IBV and RV, respectively, using enzyme linked immunosorbent assays. All 854 sera collected in the three phases were screened for NDV using hemagglutination inhibition test. In chickens 20% were seropositive to AIV, 3% to EDS'76V, 74% to IBV, 88% to NDV, and 47% to RV. The seroprevalence in flocks was 23% to AIV, 6% to EDS'76V, 79% to IBV, 89% to NDV and 56% to RV. Twenty-five percent chickens had > or = 10log(2)HI titers to NDV.