Although earlier investigators experimented with anticoccidial vaccines, the world's first commercially successful product was developed by Prof. S. A. Edgar of Auburn University, Auburn, AL. This product contained live, nonattenuated Eimeria tenella oocysts and was first marketed by Dorn and Mitchell, Inc., in 1952. Under the trade names of DM® Cecal Coccidiosis Vaccine, Coxine®, NObiCOX®, and CocciVac®, it went through several formulations containing various Eimeria species that parasitize chickens, and a further product containing turkey Eimeria species was also developed. After many product and company changes, one turkey and two chicken formulations of CocciVac® are still marketed worldwide by Schering-Plough Animal Health, Inc. Chicken and turkey formulations of Immucox®, a similar type of vaccine, were developed by Dr. E.-H. Lee and first marketed in 1985 in Canada by Vetech Laboratories, Inc.

In 1974, Dr. T. K. Jeffers of Hess and Clark, Inc., Ashland, OH, published his discovery of precocious lines of coccidia, which facilitated the development of the first attenuated anticoccidial vaccine. For commercial reasons, Jeffers was unable to do this himself, but this first attenuated vaccine was designed by Dr. M. W. Shirley and colleagues at the Houghton Poultry Research Station (HPRS) in the United Kingdom. The vaccine was commercially developed under license in the United Kingdom by Glaxo Animal Health Ltd. and then Pitman-Moore, Inc., and launched in The Netherlands during 1989 under the trade name Paracox®. After further changes in company ownership, two formulations for chickens are now marketed worldwide by Schering-Plough Animal Health, Inc.

Attenuation of coccidia by embryo adaptation was reported in 1972 in the United Kingdom by Dr. P. L. Long, who originally worked at the HPRS and later became a professor at the University of Georgia, Athens, GA. An embryo-adapted line of E. tenella was included with precocious lines of other species in a series of three attenuated vaccines for chickens under the trade name Livacox®, developed by Dr. P. Bedrník and launched in the Czech Republic in 1992 by Biopharm. The formulations of all other commercially available live anticoccidial vaccines for poultry are currently based upon the scientific principles established for the CocciVac®, Paracox® or Livacox® vaccines.

Maternal immunity can interfere with infectious bursal disease virus (IBDV) vaccine efficacy. The effect of maternal antibody (MatAb) on the immune response and detection of two vaccine viruses, an IBDV immune complex (cx) and IBDV-2512, was investigated. Vaccines were administered in ovo at 100 mean embryo infectious dose to commerical broiler embryos derived from young (29 wk) or old (63 wk) flocks. Chickens with low MatAb were challenged with an antigenic standard IBDV strain at 21 and 35 days post in ovo vaccination (PIOV). At 28 and 42 days PIOV, birds were euthanatized and bursa weight:body weight ratios were determined. MatAb of progeny from the 29-wk-old flock was higher (P < 0.05) than that from progeny of the 63-wk-old flock. Progeny titers declined by day 21 PIOV. Birds with MatAb vaccinated with either the IBDV-2512 or IBDV-Icx vaccine did not mount an antibody response by 21 days PIOV. The IBDV-Icx vaccination protected chickens against bursal atrophy when they were challenged at 21 and 35 days PIOV (83% and 77%, respectively). Resistance to challenge in the absence of antibody implies that cellular immunity plays a role in IBDV protection. The IBDV-2512 vaccine caused atrophy of the bursae, and, therefore, we could not examine its protection by these criteria. Neither vaccine could be detected by antigen-capture chemiluminescent enzyme-linked immunosorbent assay. Therefore, total RNA was extracted and reverse transcription (RT)–polymerase chain reaction (PCR) and nested PCR were conducted to amplify the VP2 (hypervariable) region on viral RNA collected at 3, 6, 9, 15, and 21 days PIOV. Nested PCR detected the VP2 region of both IBDV vaccine viruses at day 3 PIOV. Only IBDV-2512 RNA was detected on day 6 PIOV. However, the IBDV-Icx RNA was detected on days 9 and 15 PIOV but not on day 21 PIOV. The IBDV-Icx RNA was evident in tissues and could induce protection against challenge. This work gives further insight into the mechanism of the IBDV-Icx vaccine.

The effect of two infectious bursal disease virus (IBDV) vaccines (IBDV-immune complex [Icx] and IBDV-2512), administered in ovo, on the cell-mediated immunity of specific-pathogen-free (SPF) broilers was examined. A decrease (P < 0.05) in the T-cell mitogenic response occurred in birds vaccinated with both vaccines on days 9 and 21 post in ovo vaccination (PIOV), but an increase (P < 0.05) occurred on day 15 PIOV. The T cells from birds given the IBDV-2512 were less responsive. There were no significant differences in proportions of lymphocytes expressing CT4 CT8⁻ and CT8 CT4⁻ except on day 21 PIOV, where an increase (P < 0.05) in IBDV-2512–vaccinated birds and a decrease (P < 0.05) in percentage of CT4 CT8⁻ in IBDV-Icx–vaccinated birds was observed. There was an increase (P < 0.05) in percentage of CT8 CT4⁻ T cells on day 21 PIOV in both vaccinated groups. A decrease (P < 0.05) in B-cell percentage was observed on day 21 PIOV in birds given both vaccines. Results indicated that although humoral immunosuppression is associated with destruction of B cells (bursal atrophy), cell-mediated immunosuppression induced by these two IBDV vaccines in SPF birds was not associated with altered helper (CT4 CT8⁻) or cytotoxic (CT8 CT4⁻) subpopulations of T lymphocytes.

Mycoplasma gallisepticum infects a wide variety of gallineaceous birds including chickens, turkeys, and pheasants. Infection occurs both horizontally and vertically. Thus, control of the spread of M. gallisepticum to noninfected flocks is difficult. Continual monitoring is necessary to identify infected flocks even under the most stringent infectious control practices. Monitoring, however, is usually performed by measuring hemagglutination activity (HA) in serum, an insensitive and variable test. Variability in the HA test arises from differences in agglutination antigen, changes in antigenic profiles of the M. gallisepticum strain, and variability in reading the agglutination reaction. Enzyme-linked immunosorbent assays (ELISAs) are the preferred method of testing because of the ease in obtaining sera and the sensitivity and reproducibility of the assays, but the ELISA suffers from a lack of standardization in the test antigen. The ELISA test will be more easily accepted once the test antigen has been standardized. To this end, we have identified, cloned, and characterized the gene for an antigen that has potential as a species-specific antigen for M. gallisepticum. The gene codes for a 75-kD protein, P75, that is recognized during natural infections. Recombinant P75 is not recognized in immunoblots by convalescent sera produced in chickens infected with Mycoplasma synoviae, Mycoplasma gallinarum, and Mycoplasma gallinaceum or in turkeys infected with Mycoplasma meleagridis.

Studies have indicated variations in the degree of efficacy of certain commercial disinfectants used in poultry production facilities. We used an adequate method of in vitro testing to compare the efficacy of disinfectants while testing them in conditions similar to those of the poultry facilities. BioSentry 904, ethylenediaminetetracetic acid (EDTA)-Tris, and a combination of the two were tested by this method against five field isolates of Pseudomonas aeruginosa at 10³, 10⁶, and 10⁹ colony-forming units (CFU)/ml. At the 10⁹ CFU/ml concentration, most compounds failed to achieve a total kill with a contact time of 15 min. When tested at bacterial concentrations of 10³ CFU/ml, the combination of EDTA-Tris mixed at a 1:1 ratio with BioSentry 904 killed the bacteria upon initial contact (≤0.05 min). This disinfectant mixture exhibited antagonistic, indifferent, or synergetic effects when exposed to different bacterial isolates at a concentration of 10⁶ CFU/ml.

The spike 1 (S1) surface glycoprotein of infectious bronchitis virus (IBV) is the major inducer of the generation of virus neutralizing antibodies, and the administration of purified S1 has been shown to elicit a protective immune response against virulent virus challenge. On the basis of these observations, recombinant fowl poxvirus (rFPV) containing a cDNA copy of the S1 gene of IBV Mass 41 (rFPV-S1) was constructed and its immunogenicity and vaccine potential were evaluated. Initially, rFPV-S1 was shown to express the S1 in vitro by indirect immunofluorescence staining and western blot analyses. Later, in vivo expression was demonstrated by the detection of IBV-specific serum immunoglobulin G and neutralization antibodies in the sera of chickens immunized with rFPV-S1. That the recombinant virus elicited anti-IBV protective immunity was indicated by the manifested, relatively mild clinical signs of disease, decreased titers of recovered challenge virus, and less severe histologic changes of the tracheas in virulent IBV Mass 41–challenged chickens previously receiving rFPV-S1 as compared with parental fowl poxvirus (FPV)-vaccinated control birds. In contrast, chickens immunized with either recombinant or parental FPV were resistant to a subsequent virulent FPV challenge. As to a preferred method of immunization, wing web administration appeared to be superior to the subcutaneous route because a greater percentage of birds vaccinated by the former protocol exhibited an anti-IBV humoral immune response. Thus, rFPV-S1 has potential as a poultry vaccine against both fowl pox and infectious bronchitis.

Two trials were conducted to define temporal changes in plasma d-α-tocopherol (AT) caused by infection with Eimeria maxima in chickens that consumed either low (25 ppm) or high (225 ppm) levels of dietary dl-α-tocopheryl acetate (VE-AC) from 1 day of age. In both trials, rates of weight gain were depressed between days 5 and 7 postinoculation (PI) and were not influenced by the level of dietary VE-AC. Plasma AT was consistently depressed at 5 and 7 days PI in chickens consuming either level of dietary VE-AC. The pattern and degree of plasma AT depression correlated with those of plasma carotenoids. Plasma levels of NO₂⁻ NO₃⁻ were significantly increased at 5 and 7 days PI. In trial 1, the average increase during that period was not as high in chicks consuming 225 ppm VE-AC, but in trial 2, diet had no effect on the degree of increase. Also, there were no consistent effects of dietary VE-AC on lesion scores or amount of oocysts shed. These results are in general accord with findings of earlier experiments, and we conclude that feeding high levels of VE-AC to broiler chicks from 1 day of age is not effective in mitigating the pathology, including weight gain depression and development of mucosal lesions, during E. maxima infections or in modifying immune response events associated with phagocytosis as indexed by plasma NO₂⁻ NO₃⁻. The likely basis for the ineffectiveness of feeding this fat-soluble form of vitamin E is that it is malabsorbed during E. maxima infection in the same manner as carotenoids and becomes less biologically available to infected tissues during the acute phase of infection.

Nephropathogenic infectious bronchitis (NIB) was diagnosed in 28 infectious bronchitis virus (IBV)–vaccinated commercial chicken flocks in Pennsylvania from December 1997 to July 2000. Early clinical signs were increased flock mortality and urinary water loss (polyuria and pollakiuria) leading to wet litter. Daily mortality ranged from 0.01% in layers to 2.45% in broilers, with total broiler mortality as high as 23%. Severe renal swelling and accumulation of urates in the tubules were commonly seen. Visceral gout and urolithiasis were less frequently observed. Histopathologic changes included characteristic tubular epithelial degeneration and sloughing with lymphoplasmacytic interstitial nephritis. Minimal respiratory disease signs were noted in broilers. Egg production and shell quality declined in layers.

Confirmatory diagnosis of NIB was made by IBV antigen–specific immunohistochemical staining of the renal tubular epithelium and virus isolation. Sequencing of the S1 subunit gene of 21 IBV isolates showed the NIB outbreak to be associated with two unique genotypes, PA/Wolgemuth/98 and PA/171/99. The cases from which the genotypes were isolated were clinically indistinguishable. The NIB viruses were unrelated to previously recognized endemic strains in Pennsylvania and were also dissimilar to each other. Genotype PA/Wolgemuth/98 was isolated almost exclusively during the first 14 mo of the outbreak, whereas PA/171/99 was recovered during the final 18 mo. The reason for the apparent replacement of PA/Wolgemuth/98 by PA/171/99 is not known.

Nine Spanish isolates of infectious bursal disease virus (IBDV) were characterized and classified after reverse transcriptase–polymerase chain reaction of a 248-bp fragment of the VP2 gene hypervariable region and restriction fragment length polymorphism (RFLP). The restriction endonucleases (REs) used were BstNI, SacI, SspI, TaqI, DraI, and StyI. Sequencing of the amplified product and further comparison of these sequences with published sequence data from other IBDV strains were also performed. Very virulent and classic strains were identified. None of the strains identified had molecular characteristics similar to that of the American variant strains. Four very virulent strains (VG-248, 5939, 6145, and 7333) were digested by the TaqI, SspI, and StyI enzymes. The sequences of these strains were closely related to other European and Japanese very virulent IBDV (vvIBDV) strains. Strains VG-311, VG-262, and VG-208 were digested by the BstNI and SacI REs and were classified as classic strains. Strains VG-276 and VG-313 had unique RFLP patterns. VG-276 exhibited the SspI RE site, which has been reported as a characteristic of vvIBDV strains, whereas the VG-313 strain exhibited a SacI and StyI RE site indicative of the classic IBDV Edgar and 52–70 strains. However, nucleotide sequence analysis of the amplified hypervariable region strain VG-276 revealed a higher identity with the classic strains STC, 52/70, and 9109 IBDV strains, whereas strain VG-313 exhibited a higher identity with the vvIBDV strains.

Although previous studies have demonstrated an association between interferon-gamma (IFN-γ) promoter genotype and antibody response kinetics in chickens, the protein levels that may mediate such a gene–trait association have not been determined. The objective of this study, therefore, was to determine the correlation of circulating IFN-γ levels with both the IFN-γ promoter polymorphisms and antibody response in order to evaluate the potential role of IFN-γ protein in mediating genetic control of antibody response in chickens. Antibody response after Salmonella enteritidis (SE) vaccination at day 10, antibody response to sheep red blood cells (SRBCs) and killed Brucella abortus after immunizations at 19 wk and 22 wk, and serum IFN-γ protein level were measured in an F₂ population derived from inbred lines. A single nucleotide polymorphism in the IFN-γ promoter region was associated with IFN-γ protein expression as measured by an enzyme-linked immunosorbent assay after both primary and secondary immunizations. Higher IFN-γ protein level was correlated with higher antibody level to SE and with increased maximum level and decreased time to reach the maximum secondary antibody response to SRBCs. These results suggest that one of the mechanisms by which promoter polymorphism of IFN-γ affects antibody production in chickens may involve the circulating level of IFN-γ protein.

This investigation detailed the clinical disease, gross and histologic lesions, and distribution of viral antigen in juvenile laughing gulls (Larus atricilla) intranasally inoculated with either the A/tern/South Africa/61 (H5N3) (tern/SA) influenza virus or the A/chicken/Hong Kong/220/97 (H5N1) (chicken/HK) influenza virus, which are both highly pathogenic for chickens. Neither morbidity nor mortality was observed in gulls inoculated with either virus within the 14-day investigative period. Gross lesions resultant from infection with either virus were only mild, with the tern/SA virus causing decreased lucency of the air sacs (2/6), splenomegaly (2/6), and pancreatic mottling (1/6) and the chicken/HK virus causing only decreased lucency of the air sacs (2/8) and conjunctival edema (2/8). Histologic lesions in the tern/SA-inoculated gulls included a mild to moderate heterophilic to lymphoplasmacytic airsacculitis (6/6), mild to moderate interstitial pneumonia (3/6), and moderate necrotizing pancreatitis and hepatitis at 14 days postinoculation (DPI) (2/6). Immunohistochemical demonstration of viral antigen occurred only in association with lesions in the liver and pancreas. In contrast, viral antigen was not demonstrated in any tissues from the chicken/HK-inoculated gulls, and inflammatory lesions were confined to the air sac (3/8) and lungs (3/8). Both viruses were isolated at low titers (<10^1.68 mean embryo lethal dose) from oropharyngeal and cloacal swabs up to 7 days postinoculation (DPI), from the lung and kidney of one of two tern/SA-inoculated gulls at 14 DPI, and from the lung of one of two chicken/HK-inoculated gulls at 7 DPI. Antibodies to influenza viruses as determined with the agar gel precipitin test at 14 DPI were detected only in the two tern/SA-inoculated gulls and not in the two chicken/HK-inoculated gulls.

Under simulated natural conditions of bird production and parasite challenge, the effects of ascaridiasis and the effectiveness of fenbendazole treatment (6-day regimes in the feed at 16 ppm) were documented. Birds were artificially challenged with ascarid larvae on a daily basis from day 35 to 112, with bird grow out ending on day 119. Experimental groups, on a per pen basis, were infected control, treated with fenbendazole at days 63–69, treated with fenbendazole at days 63–69 and days 91–97, and uninfected control. In the same order as above, and on an experimental group mean bird basis, final weights were 13.34, 13.47, 13.59, and 13.78 kg, average daily gains from day 7 to day 119 were 117.8, 118.9, 120.1, and 121.8 g, and units gained per unit of feed consumed from day 7 to day 119 were 0.337, 0.341, 0.347, and 0.362. Infected control bird mean Ascaridia dissimilis burdens, with all stages combined, ranged from 351.1 on day 63 to 117.2 on day 91, levels seen commonly with naturally infected commercial turkeys. Trial data clearly indicated that moderate A. dissimilis burdens negatively impacted animal performance (average daily gains and feed efficiencies) and that these parasite burdens are effectively removed by fenbendazole treatment.

The pathogenicity of serotype 1 group I avian adenovirus (GIAAV) strains isolated from gizzard erosions for 1-wk-old specific-pathogen-free (SPF) chickens was investigated. In Experiment 1, SPF chickens inoculated orally with a serotype 1 strain (Tokushima2000/GE) of GIAAV isolated from gizzard erosion in broilers were euthanatized and necropsied from 3 to 12 days after inoculation. Clinically, there were no significant signs in the chickens inoculated with Tokushima strain. The gizzards were found macroscopically to be dilated with liquefied contents. The koilin layer of the gizzard exhibited multifocal white lesions with occasional erosions. Gross lesions of the koilin layer (white foci and erosion) appeared at 3 days after inoculation and persisted until 12 days after inoculation. The gross lesions were most severe at 9 days after inoculation. Histologically, multifocal degeneration, necrosis, and loss of the glandular epithelial cells with intranuclear inclusions, liquefactive change of koilin substance, and erosive lesions of koilin layers and surface epithelium of the gizzards were observed. Macrophages increased in the focal areas of the lamina propria below the liquefactive koilin substances. Immunohistochemically, intranuclear inclusions were positive against GIAAV antigen. Histologic lesions of gizzards were most severe at 7 and 9 days after inoculation, but the frequency of intranuclear inclusions was greatest at 5 days after inoculation. In Experiment 2, the pathogenicities of four strains of serotype 1 GIAAV for 1-wk-old SPF chickens were compared. Three strains isolated from gizzard erosion had similar pathogenicities but one strain from a normal chicken was not pathogenic for chickens. These results suggest that gizzard erosions by serotype 1 GIAAV infection may occur in broiler chickens in Japan.

Floor pen studies were conducted, with broilers from 1 to 7 wk of age and with turkeys from 1 to 14 wk of age, to evaluate the chronic effects of moniliformin (M). Fusarium fujikuroi (M-1214) culture material was added to typical corn-soybean basal diets to supply 0, 25, or 50 mg M/kg diet (broilers) or 0, 12.5, 25, 37.5, or 50 mg M/kg diet (turkeys). Compared with controls, chicks fed diets containing 50 mg M/kg consumed more feed, had lower body weight gain, were less efficient in converting feed to body weight gain, and had increased relative heart and proventriculus weights. Chicks fed the diet containing 50 mg M/kg also had significantly higher mortality and decreased mean corpuscular volumes compared with controls. Broilers fed 25 and 50 mg M/kg also had increased serum gamma glutamyltransferase activities. Feed intake, body weight gain, and feed conversion of turkeys fed dietary M were not affected. At 6 and 14 wk, turkeys fed 25, 37.5, or 50 mg M/kg diet had increased (P < 0.05) relative heart weights when compared with controls. At week 14, turkeys fed diets containing 37.5 or 50 mg M/kg also had increased (P < 0.05) relative liver weights compared with turkeys fed 0, 12.5, or 25 mg M/kg diet. Lesions, observed only in the hearts of broilers and turkeys fed 50 mg M/kg, were loss of cardiomyocyte cross striations, increased cardiomyocyte nuclear size, and an increased number of cardiomyocyte mitotic figures (turkeys only). Results indicate that ≥37.5 mg M/kg is hepatoxic and ≥25 mg M/kg is cardiotoxic to turkeys and 50 mg M/kg diet is toxic to broilers fed to market age.

A statistical approach was used to establish a new classification system of Marek's disease virus (MDV) on the basis of neurologic responses. To develop the system, neurologic response data from 15×7 chickens inoculated with 30 strains of serotype 1 MDV were statistically analyzed by a cluster analysis. The goal was to identify a statistical system that would verify if three neurovirulence groups correlated with the three pathotypes previously described. The system was also validated in two additional strains of specific-pathogen-free (SPF) chickens, SPAFAS and line SC (Hy-Vac).

The proposed system is based on analysis of three variables: 1) frequency of birds showing transient paralysis between 9 and 11 days postinoculation (dpi), (2) mortality before 15 dpi, and (3) frequency of birds showing persistent neurologic disease between 21 and 23 dpi. By use of this system, a MDV may be classified in one of three groups, designated neuropathotypes A, B, and C, which roughly correspond to the virulent, very virulent, and very virulent plus pathotypes, respectively. However, correlation between neuropathotype and pathotype was not absolute, and neuropathotyping is more a complement to the current pathotyping system than a replacement for it. Our results showed that neuropathotyping studies can be conducted in two types of commercial SPF chickens by the use of the same variables, although the system would first have to be standardized by the use of prototype viruses. Neuropathotypes can also be estimated without statistical analysis with reasonable accuracy. By use of this analysis, we established that MDV strains within the very virulent pathotype may be subdivided into neuropathotypes B and C, thus establishing a previously unrecognized pathotypic classification. This finding illustrates how neuropathotyping may extend important information not identified by conventional pathotyping.

Three groups of >60-wk-old broiler breeder hens were assessed for the presence of Campylobacter within segments of the reproductive tract. In the first group, after stunned, the hens were bled, scalded, and defeathered, the reproductive tracts were aseptically excised from 18 hens, six from each of three adjacent floor pens that were feces positive for Campylobacter. The reproductive tract segments (infundibulum, magnum–isthmus, shell gland, vagina, and cloaca) were pooled by pen. In the second group, 10 individual hens were sampled from the pens; the reproductive tract was divided into the following segments: magnum, isthmus, shell gland, vagina, and cloaca. For the third group, hens were obtained from two commercial farms that had been determined to be feces positive for Campylobacter, and the reproductive tract was divided into five segments, as described for the second group. Segments of the reproductive tract were placed into sterile plastic bags and suspended 1:3 (w/v) in Bolton enrichment broth, and serial dilutions were plated (0.1 ml) onto Campy-Cefex agar. The agar plates were incubated at 42 C for 24 hr in a microaerobic atmosphere. In group 1, the pooled reproductive tract segments for hens from pen A were Campylobacter positive for the shell gland, vagina, and cloaca; hens from pen B were positive for the cloaca only; and hens from pen C were positive for the magnum–isthmus and cloaca. In the second group, 9 of 10 cloaca samples were Campylobacter positive. Commercial hens in group 3 had campylobacter-positive cloaca samples (12/12), vagina (10/12), shell gland (7/12), isthmus (2/12), and magnum (4/12). Campylobacter colonization of the reproductive tract of the hen could enable vertical transmission of Campylobacter from the hen to the chick.

Studies were conducted to better understand the relationship among Marek's disease (MD) vaccine strains between induction of protective immunity and the degree of attenuation (or virulence). To obtain viruses at different stages of attenuation, very virulent plus MD strains 584A and 648A and selected clones of these strains were serially passaged in chicken and duck cells. These viruses were considered fully attenuated after passage for 70–100 times in chicken embryo cell cultures until they no longer induced gross lesions in susceptible, maternal antibody–negative (ab−) chickens. Lower passages of the same strains were considered partially attenuated, provided their virulence was less than that of the parent strain. Four of five partially attenuated preparations derived from MD virus strains 584A and 648A or the previously attenuated Md11 strain induced 28%–62% higher levels of protection in maternal antibody–positive (ab ) chickens against virulent MD challenge than the fully attenuated counterpart viruses. The partially attenuated 584A/d2/3 strain replicated in chickens but was totally nonprotective. Data from two subsequent trials in ab chickens confirmed that protection induced by the partly attenuated (passage 80) preparations was 79% and 118% higher, respectively, than that induced by the fully attenuated (passage 100) preparations of strain 648A. However, in one trial with ab− chickens, no difference in protection between partially and fully attenuated virus was observed. Strong protection (up to 85%) against highly virulent challenge also was provided by preparations of 648A at passages 40–60, which were moderately oncogenic when used alone. Partially attenuated strains tended to replicate to higher titers in both ab and ab− chickens compared with fully attenuated vaccines. Also, ab and ab− chickens vaccinated with partially attenuated strains developed three- to nine fold more extensive microscopic lesions in peripheral nerves at 14 and 22 days after virulent challenge than chickens vaccinated with fully attenuated strains. When measured in ab chickens, loss of lesion induction by 648A was achieved 30 passages earlier (at passage 70) than when measured in ab− chickens. Thus, maternal antibodies appeared to abrogate the pathogenicity of some partially attenuated strains. These studies establish for MD the principle that at least some partially attenuated MD viruses may replicate better and induce stronger immunity against virulent challenge than fully attenuated preparations of the same strain, at least when tested in ab chickens. Moreover, depending on passage level, partially attenuated vaccine strains may be relatively innocuous for ab chickens, causing few or no lesions.

Protection provided by live and inactivated virus vaccination against challenge with the virulent nephropathogenic infectious bronchitis virus (NIBV) strain PA/Wolgemuth/98 was assessed. Vaccinations with combinations of live attenuated strains Massachusetts (Mass) Connecticut (Conn) or Mass Arkansas (Ark) were given by eyedrop to 2-wk-old specific-pathogen-free leghorn chickens. After live infectious bronchitis virus (IBV) vaccination, some chickens at 6 wk of age received an injection of either an oil emulsion vaccine containing inactivated IBV strains Mass Ark or an autogenous vaccine prepared from NIBV PA/Wolgemuth/98. Challenge with PA/Wolgemuth/98 was given via eyedrop at 10 wk of age.

Serum IBV enzyme-linked immunosorbent assay antibody geometric mean titers (GMTs) after vaccination with the combinations of live attenuated strains were low, ranging from 184 to 1354, prior to NIBV challenge at 10 wk of age. Both inactivated vaccines induced an anamnestic response of similar magnitudes with serum GMTs of 6232–12,241.

Assessment of protection following NIBV challenge was based on several criteria: virus reisolation from trachea and kidney and renal microscopic pathology and IBV-specific antigen immunohistochemistry (IHC). Live attenuated virus vaccination alone with combinations of strains Mass Conn or Mass Ark did not protect the respiratory tract and kidney of chickens after PA/Wolgemuth/98 challenge. Chickens given a live combination vaccination of Mass Conn and boosted with an inactivated Mass Ark vaccine were also susceptible to NIBV challenge on the basis of virus isolation from trachea and kidney but showed protection on the basis of renal microscopic pathology and IHC. Live IBV-primed chickens vaccinated with an autogenous inactivated PA/Wolgemuth/98 vaccine had the highest protection against homologous virulent NIBV challenge on the basis of virus isolation.

CIAV-7 is a virus with similar pathogenic and physicochemical characteristics to, but antigenically distinct from, chicken infectious anemia virus (CIAV). The pathogenesis of CIAV-7 was evaluated in a comparative study with a representative isolate of CIAV, the Del-Ros strain. The pathogenesis of CIAV-7 was similar to Del-Ros on the basis of the clinical disease induced and gross and microscopic lesions, although CIAV-7 produced fewer and less severe lesions overall. A second comparative pathogenesis study was performed with Del-Ros and CIAV-7, both alone and in combination with infectious bursal disease virus (IBDV). In this study, the pathogenesis of CIAV-7 was similar to Del-Ros in clinical, gross, and microscopic lesions in the bone marrow. However, thymic lesions were less severe in CIAV-7–inoculated birds. The interaction between Del-Ros and IBDV was synergistic, whereas there was no observed potentiation of CIAV-7–induced disease by IBDV. Progeny from breeder flocks from several geographic locations in the eastern United States were challenged with CIAV-7 or Del-Ros to assess protection by maternal antibodies. Some progeny from all flocks had protection against CIAV-7 challenge, providing evidence for the presence of CIAV-7 in the field. Additionally, the number of birds protected against CIAV-7 or Del-Ros challenge varied within flocks, demonstrating that the agents are serologically distinct.

A putative new serotype of chicken infectious anemia virus (CIAV) isolated from 17-wk-old broiler breeder pullets was compared with a known, previously characterized CIAV isolate, the Del-Ros strain. Physicochemical characteristics evaluated included thermal stability, size, pH, and chloroform sensitivity. Physicochemically, CIAV-7 was identical to CIAV. The virus isolates were compared antigenically by enzyme-linked immunosorbent assay, virus neutralization, immunofluorescence assay, and western blot. All four serologic assays demonstrated that CIAV-7 is antigenically distinct from the Del-Ros strain of CIAV. Additionally, polymerase chain reaction (PCR) and Southern blot were used to determine if there were similarities in genome sequence between the two viruses. CIAV-7 could not be detected with CIAV-specific PCR primers or a with CIAV-specific probe by Southern hybridization.

Formaldehyde administration in the hatchery can be very useful in decreasing microbial numbers. However, its use is controversial because of the adverse effects that can occur to chicks and people. This study was designed to look at alternative methods of application of formaldehyde in the hatchery. In addition, the study compared the effects of these methods of application on in ovo–and non-in ovo–injected eggs. All in ovo–injected eggs were given diluent only with no vaccine or antibiotic added. In hatchers containing both in ovo–injected eggs and non-in ovo–injected eggs, formaldehyde was administered two ways, dose (DOSE) and constant rate infusion (CRI). In the DOSE hatcher, 12 ml of formaldehyde was administered at one time every 12 hr, whereas in the CRI hatcher, the same volume was administered at a rate of 1 ml/hr over a 12-hr period. A control (CONT) hatcher received 12 ml of distilled water at the same time that the DOSE hatcher was given formaldehyde. In the DOSE hatcher, a peak concentration of formaldehyde of 102 ppm was reached. The CRI was maintained at approximately 20 ppm of formaldehyde. At pipping, the aerosol bacterial load in the hatchers receiving formaldehyde (DOSE, 130 colony-forming units [CFU]/m³; CRI, 82.5 CFU/m³) was significantly less than in the CONT hatcher (235 CFU/m³). At hatch, the CRI (337.5 CFU/m³) was not able to control bacterial levels and only the DOSE hatcher (150 CFU/m³) had a significantly lower aerosol bacterial count. The CRI non-in ovo–injected eggs (93.39%) had a significantly higher percentage of hatch of fertile compared with non-in ovo–injected eggs exposed to water (84.27%). In ovo–injected eggs in CONT and DOSE treatment groups contained significantly higher percentages of visual contamination than non-in ovo–injected eggs in the same hatchers. This difference had numerical significance only in the treatment groups within the CRI hatcher. The chicks were then placed into replicate treatment groups and grown for 14 days. Chicks from the CRI in ovo–injected eggs had a statistically significant improvement in feed conversion ratio (1.24) at 14 days when compared with chicks from CONT non-in ovo–injected eggs (1.29). All formaldehyde-exposed chicks had numerically lower feed conversion ratios compared with the CONT exposed chicks.

Feather pulp from experimentally infected chickens was used as a source of DNA for polymerase chain reaction (PCR) amplification of avian leukosis virus subgroup J (ALV-J) proviral DNA. A primer set that produces a large amplicon (∼2125) was used to detect ALV-J proviral DNA. This primer set was used in lieu of previously published primers because it allows for sequencing of the entire envelope gene and because it was able to detect diagnostically a number of North American ALV-J isolates that could not be detected with previously published primers and PCR conditions. ALV-J proviral DNA was detected in feather pulp at 7 days of age in more than 90% of birds infected as embryos and 7 days postinoculation in over 50% of chickens infected at 3 days of age. The results obtained with PCR on feather pulp were compared with those of virus isolation. In the embryo-inoculated birds, the percentages of agreement between PCR and virus isolation were 92.5% at 7 days of age and 100% at 28, 42, 49, and 56 days of age. However, the overall sensitivity of virus isolation in embryo-infected birds was higher, particularly at 7 and 56 days of age. In chickens inoculated at 3 days of age, the percentages of agreement of detection between PCR and virus isolation ranged from 75% at 10 days of age to 100% at 42 days of age. Agreement of negative results of ALV-J detection by PCR and virus isolation in chickens infected posthatch ranged between 66.6% and 100% between the ages of 10 and 42 days. Virus isolation requires chicken embryo fibroblasts of specific genetic lines, and the process takes on average 7–9 days. Aseptic collection of blood and tissues for virus isolation and molecular detection of ALV-J requires sterile necropsy instruments as well as syringes and needles for each individual chicken, whereas sterile microcentrifuge tubes and gloves are the only equipment necessary for aseptic feather pulp collection for ALV-J detection by PCR. PCR-based detection of ALV-J in feather pulp is especially suitable when ALV-J infection must be diagnosed rapidly and unequivocally without killing the chicken(s) and in situations where crucial reagents or suitable virus propagation substrates are not readily available for isolation and propagation of ALV-J in cell culture.

Unfounded field speculation has suggested that avian leukosis virus subgroup J (ALV-J) predisposes young meat-type chickens to inclusion body hepatitis caused by fowl adenovirus (FAV). To address this hypothesis, we infected 1-day-old grandparent meat-type chickens carrying maternal antibodies against FAV with a field isolate of FAV associated with inclusion body hepatitis in broilers, ALV-J, or both FAV and ALV-J. We examined the effects of FAV alone or in combination with ALV-J on the basis of clinical signs, overall mortality, growth rate, and gross and microscopic lesions. With such criteria for evaluating possible interactions, we found no significant differences in the dually infected birds in comparison with chickens that received a monovalent challenge with either FAV or ALV-J.

We have studied colonization of crops in newly hatched leghorn chicks (a layer breed) by wild-type and mutant strains of Campylobacter jejuni. We established that the wild-type parent strain forms a stable population level within the crop and that the mutant strains will do likewise. Concentrations of mutant strains in the crop were usually below that of the wild-type parent strain and ranged from 10³ to 10⁵ colony-forming units. These results differ from results we have previously reported concerning cecal colonization, where these same mutant strains lacked colonizing ability. The present results, therefore, indicate that bacterial factors necessary for colonization of the crop are not the same as those needed for colonization of the cecum.

Pulmonary hypertension syndrome (PHS), also known as ascites, in broiler chickens prevailed in the local area of Ibaraki prefecture, Japan, and was investigated epidemiologically, serologically, and pathologically. PHS developed in chickens older than 35 days of age when rapid increase of body weight started. Approximately 90% of affected birds were males, in which weight increase was greater than in females. Serologic test revealed that PHS broilers had an increase of hematocrit value. Pathologic studies indicated that the heart of affected birds had an obese-induced pressure and cold exposure triggered congestion in the right ventricle/cava and an increase in peritoneal fluid. These changes were consistent with the previous reports of PHS, so we designed the experiment of effects on cold-induced PHS birds in a temperature-controlled house. After the 10 PHS birds at 55 days were reared for 14 days in a temperature-controlled house at 20 ± 5 C, ascites disappeared in eight birds and hematocrit values decreased to normal range in nine birds. Our finding indicated that temperature-controlled environment may be one solution to reduce mortality in PHS birds.

Concern by consumers about food safety has resulted in increased pressure on poultry companies to develop effective sanitation programs. Salmonella isolates in hatcheries are often the same species isolated from processing plants. Resistance develops in bacteria after prolonged exposure to disinfectants. The methods available in published literature to detect the efficacy of disinfectants are labor intensive and do not consider how bacteria behave when adhered to a solid surface. We used a recently developed technique, which utilizes the actual surfaces on which the disinfectant is to be applied, to evaluate the degree of resistance to four commercially available disinfectants of 17 bacterial isolates from poultry hatcheries. We found that bacterial isolates within the same genus and species have different sensitivities to the same disinfectant. In addition, disinfectants with similar but not identical chemical formulations have different efficacies against the same bacteria.

The pathogenicity of serotype 2 OH strain of infectious bursal disease virus (IBDV) to specific-pathogen-free (SPF) chicken embryos and 2-wk-old SPF chickens and turkey poults was investigated. The virus was pathogenic for chicken embryos after five passages as evidenced by pathologic changes in inoculated embryos. The embryo-adapted virus was not pathogenic for 2-wk-old SPF chickens and turkey poults as indicated by lack of clinical signs, gross or microscopic lesions in the bursa of Fabricius of inoculated birds. Bursa-to-body-weight ratios of the inoculated chickens and turkey poults were not significantly different from those of uninoculated controls. Virus-neutralizing antibodies to serotype 2 IBDV were detected in inoculated chickens and turkeys. Results of this study indicated that the embryo-adapted serotype 2 OH IBDV isolate that is pathogenic for chicken embryos is infectious but not pathogenic in chickens and turkeys.

Lines of white leghorn chickens were selectively bred for either a high (H) or low (L) antibody response to sheep erythrocytes. The parental lines, HH and LL, and reciprocal crosses, HL (sire line cited first and dam line second) and LH, were compared for their responses to various diseases. High antibody titers were associated with reduced body weight. Lines and their crosses were challenged with infectious diseases. The LL line was most resistant to Mycobacterium avium, whereas the HH line was most susceptible. The HH line was most resistant to Mycoplasma gallisepticum, whereas the LL line was most susceptible. These findings indicate that defense against infectious diseases are resource expensive. In order to save resources, it is possible that different parts of a population might genetically devote high levels of resources against different types of diseases so that the entire population is not susceptible to a single infection.

Ornithobacterium rhinotracheale (ORT) has been associated with respiratory disease, increased mortality, retarded growth, and decreased egg production in chickens and turkeys. Surveillance of exposure to ORT infection in the field has shown that prevalence of the infection is higher during winter months. The ability of ORT to remain viable in the poultry litter was studied at different temperatures over time. Presterilized poultry litter was inoculated with 10¹¹ colony-forming units of ORT and kept at −12 C, 4 C, 22 C, 37 C, and 42 C. Reisolation and titration of ORT from litter was attempted at intervals. Results indicate that ORT survived for 1 day at 37 C, 6 days at 22 C, 40 days at 4 C, and at least 150 days at −12 C. ORT did not survive 24 hr at 42 C. The survival of ORT at lower temperatures may be associated with the higher incidence of ORT infection in poultry during winter months.

In this report, we show that chickens, infected with Salmonella enteritidis (SE) by oral gavage, produce secretory immunoglobulin As (sIgAs) that specifically bind to numerous SE antigens. Chickens infected with SE showed strong sIgA response against flagella in both bile and crop. The optical density values of enzyme-linked immunosorbent assay (ELISA) tests in positive bile and crop were 1.17 and 0.38, respectively, and were significantly different from those of negative samples. Western immunoblotting revealed that ∼13.5-, ∼56-, ∼62-, ∼80-, and ∼143-kD polypeptides were immunodominant proteins in bile, whereas ∼56-, ∼62-, and ∼80-kD polypeptides were found to be strong antigens in crop. These results indicate that the crop may function as another site for mucosal immunity, and the SE flagella-based ELISA of crop samples can provide a useful screening test of SE exposure in chickens.

Poult enteritis mortality syndrome (PEMS) has been an economically devastating disease in North Carolina since the early 1990s. Though much is known about the disease, many questions remain unanswered about the syndrome, including its cause, transmission of causative agent(s), and control methods. This study was designed to investigate the association between PEMS and farm management factors. A prospective longitudinal study was conducted by collecting farm data and monitoring weekly mortality in 54 commercial turkey flocks raised in PEMS-affected regions. Univariate and multivariate statistical analyses revealed that enhancing rodent control methods was negatively associated (P = 0.0228) with PEMS.

Choanal cleft swab samples from 770 wild Canada geese (Branta canadensis) and 358 blue-winged teal (Anas discors), captured for relocation or banding, were examined for the presence of avian pneumovirus (APV) RNA by reverse transcription (RT)–polymerase chain reaction (PCR) and for virus isolation. The swab samples were pooled into groups of 5 or 10.

Sixty eight of 102 (66.7%) pooled goose samples were RT-PCR positive for APV RNA. Thirteen of 52 (25.0%) pooled blue-winged teal samples were RT-PCR positive for APV RNA. APV RNA-positive samples were inoculated onto chick embryo fibroblasts (CEF) and QT-35 cells. Infectious APV was isolated from five Canada goose pooled samples in CEF and from one Canada goose pool in QT-35 cells but not from blue-winged teal.

Molecular techniques have not only made timely and accurate detection of infectious bursal disease viruses (IBDVs) possible but also have allowed the identification of viral strains. Previously, we identified a genetic marker that distinguished wild-type IBDV strains from vaccine strains of the virus. The marker was an NgoM IV restriction enzyme site in the VP2 gene that was present in 10 wild-type viruses but not 16 vaccine strains of IBDV. On the basis of that study, we concluded that the NgoM IV marker could be useful in the identification of wild-type potentially pathogenic strains of this virus. Because virulent (hot) vaccine strains of IBDV are used to vaccinate commercial poultry, it was important to determine if the NgoM IV marker was present in these virulent vaccines. The infectious bursal disease Blen and Bursa Vac virulent vaccines were examined and determined to contain the marker. We concluded that the presence of this marker was not unique to wild-type strains of the virus. The absence of the NgoM IV marker, however, was consistent with some level of attenuation, and its presence appears to be consistent with virulent IBDV strains.

In order to assess the effects of subgroup J avian leukosis virus (ALV-J) on semen quality, broiler breeder males were separated by ALV-J status (ALV-J positive = POS, ALV-J negative = NEG) at 44 wk of age. Of the 249 males originally placed at 1 day of age, 101 (40.6%) died by 43 wk of age. Observations of tumor expression and high mortality suggest that many of the males that died prior to 44 wk of age were infected with ALV-J.

From 47 to 56 wk of age, hens were inseminated every third week with 7.5 × 10⁷ sperm. Fertility and hatch data were collected by incubating eggs laid during the 2 wk postinsemination (WPI). The number of sperm that penetrated the perivitelline membrane of the ovum was determined from eggs laid on the eighth day postinsemination. Sperm mobility index (SMI) was determined at 58 and 60 wk of age from all males producing semen.

Whereas SMI and sperm hole penetration measurements indicated that the sperm quality from treatments POS and NEG were similar, fertility was significantly greater in the POS treatment during the first (89.0% vs. 79.0%) and second WPI (59.3% vs. 45.0%). However, because of numerically higher hatch of fertile from the NEG group, the percentage of hatch of eggs set was similar between groups. These data suggest that ALV-J status of caged males has no influence on sperm quality or hatchability of eggs.

Severe pododermatitis is a common lesion in turkeys, unlike in broiler chickens. Both dietary factors (soybean meal, biotin, methionine) and poor litter conditions (wet litter) are documented etiologies of footpad lesions. In the United States and United Kingdom both farm managers and processors monitor lesions. The United Kingdom reports an average footpad relative score higher than that of the United States. In both countries, incidence in toms is higher than in hens. There is no significant difference by country. Data support no consistent association with other lesions (i.e., breast blisters or condemnations), breed, or performance parameters (i.e., weight gain or age). The litter type used in the United Kingdom is either straw or wood shavings, whereas in the United States producers predominantly use wood shavings. In the United Kingdom, diets are formulated with a higher protein, which affects the excreta and subsequent litter conditions. British United Turkey is the predominate breed in the United Kingdom. In the United States, three breeds, British United Turkey of America, hybrid, and Nicholas, are common. Recent research has demonstrated the association between biotin levels and pododermatitis. Factors associated with pododermatitis (such as production systems, wet litter, and stocking density) are discussed. There are some indications that increased stocking density is associated with pododermatitis lesions. Further research is needed to identify what management factors are associated with pododermatitis.

In mid-2000, a broiler chicken company in Alabama experienced high early mortality rates in chicks from two different hatcheries. Five isolates of Pseudomonas aeruginosa, obtained from these contaminated hatcheries and resulting broiler chicks with omphalitis, were selected to determine virulence of the bacteria. One-day-old specific-pathogen-free white leghorn chicks were placed into positive pressure isolation units (10 chicks per unit); feed and water were provided ad libitum. The five isolates of P. aeruginosa (1 × 10¹ or 1 × 10² colony-forming units/bird) were used to challenge two replicates of 10 chicks via yolk sac inoculation. Two control groups were injected with 0.1 ml of phosphate-buffered saline, and two groups received no treatment. Mortality was recorded daily, and the chicks that died were necropsied and liver and yolk sacs were cultured. After 14 days, the remaining chickens were euthanatized and necropsied. Bacterial isolates retrieved from liver and yolk sacs were identified by the API 20 NE typing system to confirm that they were the same as the challenge isolate. Virulence varied greatly among the isolates, resulting in mortality rates from 0 to 90%. The challenge isolates produced different and often distinctive postmortem lesion patterns. Antibiotic sensitivity tests showed that all five isolates were resistant to sulfisoxazole, ceftiofur, penicillin, lincomycin, bacitracin, oxytetracycline, erythromycin, naladixic acid, and tetracycline. The isolates varied in sensitivity to other antibiotics, but all isolates were sensitive to gentamicin.

We report isolation of a strain of fermentative coryneform bacteria from an outbreak of polyarthritis in chickens. This strain could not be assigned to any recognized bacterial taxon because its peculiar phenotype is not yet reported. The strain possessed phenotypic characteristics and fatty acid profile similar to Erysipelothrix but, on the other hand, exhibited temperature-dependent motility like Listeria. We found no evidence of either Mycoplasma synoviae or Chlamydia infection. Details of the phenotype and fatty acid profile of the isolate and measures undertaken to contain the outbreak have been described.

This report describes an outbreak of tuberculosis in a flock of 38,500 48-wk-old layer hens. Clinical characteristics of the process included a gradual drop in egg production and feed intake, as well as an increased mortality rate. Two well-defined clinical features were observed. On the one hand, a number of birds displayed good body condition and continued to lay but presented granulomatous nodular lesions, particularly in the infraorbital sinus, liver, and intestine. In contrast, other hens were emaciated and presented granulomatous lesions of various sizes throughout the internal organs. The lesions observed in the various organs of all the affected birds consisted of granulomas containing acid/alcohol-fast bacilli. The presence of Mycobacterium avium was confirmed through polymerase chain reaction techniques. This report describes the epidemiologic and histopathologic characteristics of the outbreak.

A 17-yr-old pet female peach-fronted conure (Aratinga aurea) was presented with the chief complaints of mild lethargy and weight loss with increased appetite. Antemortem diagnostics included complete blood count, plasma biochemistry, and radiography. Abnormal findings included elevated inflammatory parameters (hyperfibrinogenemia) and a space-occupying mass in the region of the liver. Histologic examination of a liver biopsy sample indicated bile duct hyperplasia leading to a presumptive diagnosis of hepatoxicosis. The bird initially showed moderate improvement with supportive care, but its condition declined 9 days after the liver biopsy. Supportive care was attempted a second time, but the bird did not improve and euthanasia was elected. Abnormal gross necropsy findings were confined to the liver, which contained multiple tan nodules that exuded yellowish fluid on cut section. Histopathologic examination revealed multicentric bile duct hyperplasia and cholangiocarcinoma as well as segmental papillary hyperplasia and adenocarcinoma in the proventriculus, ventriculus, and throughout the intestinal tract. This is the first report of concurrent internal papillomatosis, gastrointestinal adenocarcinoma, and cholangiocarcinoma in a peach-fronted conure.

A 4.5-yr-old rooster was presented for a “lump on the neck.” The bird exhibited dyspnea immediately prior to death. Necropsy revealed a crateriform lesion in the crop. Histologically, the lesion was diagnosed as an invasive squamous cell carcinoma, extending almost the entire thickness of the crop. Polymerase chain reaction and ultrastructural examination failed to identify a viral component. Early vascular invasion was identified.