Vaccination against infectious anemia of poultry (CAA)--results of field studies]

Infectious anemia of poultry is a disease of high economical significance. Connatal infection of chicks with the chicken anemia agent (CAA) via the embryonated egg causes anemia along with severe immunosuppression, thus rendering the chicken susceptible for secondary infections. In order to prevent infection of young chicks, it is necessary to induce immunity against CAA in parent flocks, with the aim to prevent connatal spread of the infection and provide maternal protection for baby chicks. In this publication, the efficacy and use of a live CAA vaccine is reported. From autumn 1986 until summer 1990, 3 experimental vaccine charges were applied in 85 broiler parent flocks with totally 3.1 million chickens. In addition, totally 293,000 broiler breeder and 171,000 layer breeder chicken were vaccinated in 1989/90. The vaccine was administered between the 13th and 19th week of life by drinking water without adverse effect to the birds. Chicken anemia symptoms were observed only at the begin of laying period in two parent flocks. These flocks had been vaccinated in the 17th and 19th week, respectively. The offsprings of all other vaccinated parent flocks remained free of chicken anemia. Day-old chicks derived from vaccinated parent flocks were protected against CAA challenge infection. It is emphasized, that vaccination should be performed within the 13th to 15th week of life, because according to our observations, this will lead to an immediate seroconversion.     

Increasing incidence of anemia among clinically ill Georgia broilers: 1988-89 vs. 1990.

The incidence of anemia in clinically ill Georgia broilers climbed from 66.4% (324/488) during 1988-89 to 80.9% (531/656) during 1990. The incidence of polycythemia fell from 1.6% (8/488) during 1988-89 to 1.5% (10/656) during 1990. Specifically, compared with 1988-89, the 1990 incidence of anemia increased significantly in chicks at age 7 days (P = 0.0002) and 28 days (P = 0.05). We have no certain explanation for this shifting incidence of anemia in clinically ill Georgia broilers. Anemic chicks have plasma that contains virus particles with morphologic characteristics consistent with a virus (chicken anemia agent [CAA]) known to cause anemia in chickens. If CAA is the predominant etiology for anemia in clinically ill Georgia broilers, then our observation could be easily explained. The increasing rate of anemia could indicate a decline in broiler health over time.     

Comparisons of packed cell volumes (PCVs) from so-called chicken anemia agent (CAA; a virus)-free broilers to PCVs from CAA-free specific-pathogen-free leghorns.

Packed cell volumes (PCVs) from 3-, 7-, 14-, 21-, 28-, and 35-day-old clinically healthy chicken anemia agent (CAA)-free and specific-pathogen-free (SPF) leghorn chicks were compared with PCVs from age-matched clinically healthy CAA-free broiler chicks. The PCVs of the SPF chicks regressed significantly (F = 20.6, df = 2/3, P < 0.025) on age in a linear fashion. The PCVs of the broiler chicks regressed significantly (F = 9.56, df = 2/3, P < 0.05) on age in a cubic parabola. The mean PCVs of the broiler chicks were significantly (P < 0.05) higher than PCVs of SPF chicks at every tested time interval. Results indicate that PCV values are higher in broiler chicks than in SPF leghorn chicks, and that PCVs increase as chicks age. Clinicians, diagnosticians, and investigators who intend to work with chicken blood must be aware of these differences.     

The effects of age, route of exposure, and coinfection with infectious bursal disease virus on the pathogenicity and transmissibility of chicken anemia agent (CAA)

Specific-pathogen-free (SPF) chickens were inoculated with several different concentrations of chicken anemia agent (CAA) by the intra-abdominal, intratracheal, or oral routes. Based on lowered hematocrit values, the birds were most susceptible to CAA introduced by the intra-abdominal route. When SPF chickens were infected with infectious bursal disease virus (IBDV) at 1 day of age, they remained susceptible to CAA up to at least 21 days, whereas birds inoculated with CAA alone were susceptible only at 1 day of age. Infectious bursal disease virus introduced at 1 day of age also increased the susceptibility of birds to contact infection with CAA and resulted in increased mortality rates in CAA inoculates. The response of SPF birds to CAA infection varied following exposure at 1 day of age to two different strains of IBDV (STC and Variant-E). Chicken anemia agent contacts and inoculates infected with the Variant-E strain were affected 1 week earlier by CAA than by STC inoculates, as evidenced by depressed hematocrits. However, the total number of birds affected was similar for both the Variant-E and STC-inoculated chickens. Commercial broiler chickens inoculated at 1, 7, 10, and 14 days of age by non-parenteral routes with CAA or a combination of CAA and IBDV had mean hematocrits that were lower than controls. Several CAA-inoculated birds were considered anemic, with hematocrit values of 25 or less, while uninoculated birds remained within normal ranges.     

Experimental studies on Yersinia enterocolitica infection in chickens exposed at 1-day old

1. Seventy 1-d-old broiler chicks were experimentally inoculated orally with Yersinia enterocolitica serotype 0:3 (1.4 x 10(11) cells/chick), 0:8 (1.6 x 10(11) cells) and 0:9 (8.0 x 10(10) cells) with or without sodium bicarbonate solution (10 g/l). 2. None of the chicks showed any overt clinical signs or pathological lesions although the organism was demonstrated in the ileum and shedding was observed up to 13 d after exposure. 3. The serotype, dose of Y. enterocolitica and administration of NaHCO3 solution had no significant effect on the weight gain of exposed broiler chicks. 4. Y. enterocolitica was isolated from the liver, spleen, heart and gall bladder of infected chicks 70 d after exposure. 5. Although broiler chicks appear resistant to high doses of Y. enterocolitica by the oral route, detection of the organism in the organs of infected chickens is of public health significance.     

The isolation and characterization of chicken anemia agent (CAA) from broilers in the United States

A chicken anemia agent (CAA) isolated from commercial broilers in the United States was characterized in vivo and in vitro. When inoculated into susceptible 1-day-old chickens, the agent induced a severe bone marrow aplasia, thymic atrophy, multiple subcutaneous and intramuscular hemorrhages, and anemia, as evidenced by reduced hematocrits. Chickens derived from different breeder flocks and inoculated in ovo or at 1 day of age varied in their susceptibility to the CAA, with some flocks being highly susceptible, while others were almost totally resistant. This was true for both specific-pathogen-free and commercial chickens. The isolate was able to pass through a 50-nm-pore-size filter and was resistant to inactivation at 56 C for 30 minutes. It failed to agglutinate avian and mammalian erythrocytes and could not be propagated in conventional cell cultures. The physical and biological characteristics of the agent and the disease it induces indicate that it is similar to the CAA found in Japan and Europe.     

Pathogenicity of CL-1 chicken anemia agent.

Twelve-day-old broiler-type chickens had hemorrhagic necrotic wing tips. After 10 blind subcultures in an MDCC-MSB1 cell line, a virus (so-called chick anemia agent [CAA]) was isolated and designated CL-1 CAA. Five-day-old specific-pathogen-free chicken embryos from a commercial breeder flock that were found not to possess antibody against CAA were infected with CL-1 virus via yolk-sac injection. Many (49%) infected embryos were small and apparently had died from severe systemic hemorrhage. Hatched chicks were small and had pale feathers, skin, skeletal muscles, bone marrow, and viscera. All infected chicks had small thymuses. These thymuses often were so small that they could not be found grossly (P = 0.002). Anemia occurred within 4 days post-hatch. Microscopically, all hematopoietic organs were markedly atrophic. Septic necrotizing lesions were seen only in organs from CL-1-injected chicks. Physicochemical and pathological characteristics of this virus indicate that it is similar to other isolates of CAA found in Europe and Japan.     

Stunting syndrome in broilers: effect of electrolytes in drinking water on performance and intestinal glucose transport

Posthatched naive or inoculated male broiler chicks were kept in separate rooms. An inoculum was prepared from intestines of stunting-syndrome affected broiler chicks. Tap water was supplied from 2 L cups, 1 cup per pen. In the Ist experiment, the naive chicks were provided with tap water only and the inoculated ones had free access to tap water or to an electrolyte solution. In the 2nd experiment, the naive and inoculated birds had free access to water in addition to an electrolyte solution. Supplementation was provided up to 3 weeks of age; thereafter all chicks had access to tap water only. Water or electrolyte consumption and body weight (BW) were determined. Total water intake of inoculated chicks was higher than that of naive counterparts (P<0.001). Electrolyte supplementation increased drinking (P<0.001) in inoculated birds more than in naive ones. At 1 week old the weight of the inoculated birds was about 64% of the weight of naive ones; at the age of 4 and 6 weeks it was about 74% and 86% respectively. Compensatory growth was most apparent in the inoculated chicks provided with electrolyte solution. At the age of 6 weeks, the latter exceeded the BW of the exclusively water supplied counterparts by 327 g. Electrolyte supplementation up to the age of 3 weeks had no effect on the naive counterparts. Osmolality was reduced slightly, but very significantly by inoculation; electrolyte supply had no effect on this variable. Sodium concentration in the plasma was higher in the inoculated birds. Plasma albumin was markedly reduced by inoculation on weeks 1 and 2. Whereas the inoculated chicks supplied with electrolytes resumed the level plasma albumin level of the naive chicks on week 3, an over-compensation occurred in the inoculated-water-supplied (IW) group, and they surpassed the naive chicks significantly. Blood hematocrit increased significantly with age; inoculation, age and/or electrolyte supplementation had no effect on this variable. Sodium-dependent glucose transport rates were enhanced in vesicles obtained from inoculated chicks as compared to naive ones. While electrolyte supplementation had no effect on glucose active transport in naive chicks, electrolyte supplementation decreased rates of glucose active transport in inoculated ones. These data demonstrate that electrolyte supplementation during the early age may be used to enhance the tolerance of broiler chicks to stunting-syndrome by improving food and water consumption, and subsequently growth rate during and after cessation of electrolyte supply.     

Diagnoses of infections by the so-called chick anemia agent: anemia and direct transmission electron microscopic detection of virus.

Blood from 48 chicks was examined for anemia (packed cell volume), and plasma was examined for virus particles by direct transmission electron microscopy (DTEM). There was agreement between the occurrence of anemia and the presence of CAA virus particles in plasma from anemic chicks (Kappa = 0.2425, Z = 2.096, P = 0.036). Although DTEM is a method that can be used to diagnose CAA in chicks, more sensitive, economical and less laborious diagnostic assays are needed.     

Isolation and identification of chicken infectious anemia virus in Brazil.

Seven chicken infectious anemia virus (CIAV) isolates were obtained from seven broiler flocks with poor performance in two states of Brazil. All isolates induced thymus atrophy, bone-marrow aplasia, and low hematocrit values when inoculated into 1-day-old susceptible chicks. The CIAV isolates were resistant to treatment with chloroform and were able to pass through 50-nm-pore-size filters. CIAV-specific antigens could be demonstrated in tissues of experimentally infected chicks using a monoclonal antibody specific for CIAV. These characteristics of the virus and the virus-induced lesions demonstrate that CIAV is present in Brazil and that the virus is associated with production problems.     

Experimental infection in specific-pathogen-free chicks with avian reovirus and avian nephritis virus isolated from broiler chicks showing runting syndrome

Avian reovirus (ARV) and avian nephritis virus (ANV) were individually isolated from runty 10-day-old broiler chicks. The ARV isolate, IR-R, the ANV isolate, IR-N, and the reference strain of ANV, G-4260, were inoculated orally into 1-day-old chicks of two specific-pathogen-free (SPF) chicken lines, 151 and PDL-1. Growth retardation without the presence of gross lesions was clearly observed at 7 and 14 days postinoculation (PI) in chicks of both lines inoculated with the IR-R strain. On the other hand, in chicks inoculated with IR-N strain, growth retardation was observed only in the chicks of line 151 at 7 and 14 days PI. Microscopically, nephritis was observed in both chicken lines at 7 and 14 days PI. When chicks that were inoculated with the IR-N strain at 1 day of age were inoculated with the IR-R strain at 3 days of age, growth retardation was observed in the chicks of line PDL-1 at 10 and 17 days PI. However, the growth retardation was less severe than in the group receiving a single inoculation of the IR-R strain.     

Effects of chicken anemia agent on lymphokine production and lymphocyte transformation in experimentally infected chickens.

One-day-old chicks with no maternal antibodies to chicken anemia agent (CAA) were inoculated intramuscularly with CAA grown in MDCC-MSB1 cells. A control group of birds from the same source was inoculated intramuscularly with a lysate from uninfected MSB1 cells. Birds were killed at 8, 15, 22, 29, and 43 days postinoculation (PI), and the spleens were removed. Spleen cells were dispersed and stimulated with various concentrations of Concanavalin A (Con A), and lymphocyte transformation responses were determined. Supernatants from Con A-stimulated cultures were assayed for T-cell growth factor (TCGF) and interferon. Decreased lymphocyte transformation and TCGF production were demonstrated at 8 and 15 days PI. This was followed by a stimulation in activities before a return to control levels at 43 days PI. Interferon levels were elevated 8 days after infection. This was followed by a significant decrease in activity compared with controls at 15, 22, and 29 days PI, and a return to control levels by 43 days PI. The results suggest that CAA infection in young chickens can produce a dramatic decrease in immune competence, which, although transitory, is likely to seriously compromise the ability of birds to mount a successful immune response to invading pathogens.     

Chicken infectious anemia in Mexico: virus identification and serology survey.

Three chicken infectious anemia (CIA) virus strains were isolated from 10 different sick broiler and replacement chicken flocks with the MDCC-MSB1 cell line. One-day-old specific-pathogen-free chicks were inoculated later, with the three original samples being positive in tissue culture; one induced signs and lesions, another only lesions typical for CIA. One isolate was selected for further trials and showed resistance to chloroform and heat (75 C for 5 min) and passed through a 45-nm filter membrane but did not pass through the 22-nm filter. These characteristics were similar to the Del Rose reference strain of chicken anemia virus. By electron microscopy, the diameter of particles obtained from the pellet of infected cell cultures was between 22 and 27 nm. Serology survey carried out with 580 serum samples from different poultry farms all over the country with a commercial enzyme-linked immunosorbent assay kit gave proof of widespread seroconversion, indicating that CIA should be considered endemic to Mexico.     

Effect of ochratoxin A on Escherichia coli-challenged broiler chicks

A study was conducted to evaluate the effects of ochratoxin A (OA) on Escherichia coli-challenged broiler chickens. Day-old broiler chicks were separated into two groups of 92 chicks each, with one group fed a control mash diet, and the other fed a mash diet containing 2 ppm OA. On day 14, each group was further separated into two groups, with one group inoculated with E. coli O78 (1 x 10(7) colony-forming units/0.5 ml), whereas the other group was not inoculated with E. coli. After E. coli inoculation on day 14, four birds from each group were euthanatized at 1, 2, 3, 5, 7, 10, 14, and 21 days postinoculation. Escherichia coli infection caused dullness, depression, huddling, and diarrhea. Mortality was 14.3% in chicks infected with E. coli but fed no OA. Mortality increased to 35.7% in chicks fed OA and infected with E. coli. Decreased body weight and reduced feed intake were observed in chicks fed OA, and the effects were more pronounced in chicks fed OA and infected with E. coli. Increased serum levels of aspartate aminotransferase, alanine aminotransferase, uric acid, and creatinine and decreased levels of total proteins, albumin, globulins, calcium, and phosphorus were observed in OA-fed birds. Escherichia coli infection did not cause significant alteration in any of the serum biochemical parameters. The presence of OA in poultry rations increased mortality and the severity of an E. coli infection.     

Biological characterisation of Australian isolates of chicken anaemia agent

Three Australian isolates of chicken anaemia agent (CAA) resisted treatment at 70 degrees C for 5 min and chloroform treatment. Although minor antigenic differences were detected using monoclonal antibodies to CAA, the Australian isolates were indistinguishable from the reference Cux-1 and Gifu-1 isolates in cross-immunofluorescence and cross-neutralisation tests employing polyclonal chicken antiserums. The Australian viruses were pathogenic for intramuscularly inoculated 1-day-old SPF chicks, but were less pathogenic for 7-day-old chicks. Thus the Australian isolates of CAA did not differ significantly in these properties from previously characterised CAA isolates from other continents.     

Economic effects of clinical chicken anemia agent infection on profitable broiler production.

An outbreak of anemia dermatitis syndrome caused by chicken anemia agent (CAA) occurred in 15 broiler flocks. An average of 29% of chickens in these flocks were derived from a common breeder flock. The breeder flock had no antibody to CAA at 20 weeks of age but had seroconverted by 31 weeks. Diseased broiler flocks were derived from eggs laid by the breeder flock between 25 and 30 weeks of age. CAA infection in the breeder flock was subclinical, with no apparent effects on mortality or performance. A strategic program of therapeutic and/or prophylactic antibiotic therapy was begun in affected broiler flocks as soon as the disease was diagnosed. Nevertheless, when the cost of therapy was taken into account, affected broiler flocks had a net income 17.3% to 19.6% lower than normal flocks. Average bird weights were 3.3% to 3.5% lower in affected flocks than in unaffected flocks, and affected flocks had a significantly greater proportion of lighter birds. Average mortality in affected flocks was 2.0% to 2.3% higher than in normal flocks, with peak mortality occurring in the third week of life. There was no apparent effect on feed-conversion ratio.     

A comparative study of the pathogenesis of malabsorption syndrome in broilers

Five malabsorption syndrome (MAS) homogenates from The Netherlands and Germany were used to reproduce MAS in broilers. We studied the histopathology after inoculation of 1-day-old broiler chicks and the agents that might be involved. Generally, the MAS homogenates induced signs that differed in severity and pathobiology. We could distinguish and classify the inoculated groups best by histopathology: proventriculitis, lesions in the small intestines in combination with proventriculitis, or lesions of the small intestines only. Lesions in the small intestine had more impact on weight gain depression than lesions in the proventriculus. In three out of five inoculated groups, microscopic lesions of the pancreas were found. Reovirus was detected in the inoculated groups by virus isolation and seroconversion, and reoviral antigen was detected by immunohistochemistry of the small intestine. Also, enteroviruslike particles were detected in three of the five inoculated groups, although not in the most affected group. Additionally, bacteriophages and bacteria (hemolytic Escherichia coli, Pasteurella hemolytica, and Enterococcus durans) were isolated from inoculated chicks. The role these agents play in pathogenesis of MAS is still unsolved.     

Incidence of anemia and polycythemia in clinically ill Georgia broilers.

The incidence of anemia and polycythemia was established in clinically ill Georgia broilers that were tested for packed cell volume (PCV) during 1988 and 1989. More than 66% (324/488 = 66.4%) of PCV-tested broiler chicks were anemic, and less than 2% (8/488 = 1.6%) of PCV-tested chicks were polycythemic. The incidence of anemia was significantly (P less than 0.001) higher than expected (2.5%) at age 7 days (56.9%), 14 days (83.9%), 21 days (74.7%), 28 days (58.7%), and 35 days (57.9%). The incidence of polycythemia was significantly (P less than 0.001) higher than expected (2.5%) in 35-day-old broilers (21.1%) but was not significantly different from the expected rate in broilers at age 7 days (0%), 14 days (1.8%), 21 days (0.3%), and 28 days (4.3%). The established rates for anemia were much higher than we would have hypothesized. This led us to believe that either 1) an etiology for anemia is present in epizootic proportions in Georgia broilers, or 2) the standard method for establishing reference intervals for anemia in animals does not apply to broiler chicks.     

Isolation of avian reovirus as a possible etiologic agent of osteoporosis ("brittle bone disease"; "femoral head necrosis") in broiler chickens

Avian reovirus was isolated from intestines of 3-to-7-day-old broiler chickens with enteritis from broiler houses where osteoporosis was a problem. The virus was purified in a cesium chloride gradient (buoyant density 1.37 gm/ml) and identified as a reovirus by electron microscopy. Specific-pathogen-free (SPF) chickens and commercial broiler chickens with anti-reovirus maternal antibodies inoculated at 1 day of age with the reovirus isolate developed lesions of femoral head fractures and/or osteoporosis; reovirus could be reisolated from the bone marrow and intestinal tracts of experimentally infected SPF birds. The reovirus isolate, although isolated from intestines, induced development fo tenosynovitis lesions in SPF and commercial broiler chickens.