The ways in which antibiotics are used in poultry production have changed considerably during the past decade, mainly because of concerns about potential negative human health consequences caused by these uses. Human health improvements directly attributable to these antibiotic-use changes are difficult to demonstrate. Given that some antibiotics will continue to be used in the poultry industry, methods are needed for estimating the causal relationship between these antibiotic uses and actual animal and human health impacts. This is a challenging task because of the numerous factors that are able to select for the emergence, dissemination, and persistence of antibiotic resistance. Managing the potential impacts of antibiotic use in poultry requires more than a simple estimation of the risks that can be attributed to the use of antibiotics in poultry. Risk models and empirical studies that evaluate interventions that are capable of minimizing the negative consequences associated with specific antibiotic uses are desperately needed.

A total of 114 male chickens from three sire families of a commercial cross of White Leghorn chickens were infected with RB-1B Marek's disease (MD) virus at 21 days of age by exposing them to chickens previously inoculated with MD virus. The presence of virus in feather tips, feather pulp, and MD viral antibodies indicated all chickens became infected. The first virus-positive chickens were observed at 12 days postexposure (dpe). The frequency reached a maximum at 27 dpe and then decreased. At 80 dpe, when the experiment was terminated, no viral DNA was detected in the feather pulp of the surviving chickens (82%). Death from MD was first observed at 38 dpe and reached 18% by the end of the experiment, with spleen lesions being the major MD lesion. The viral genome titers in spleen extracts of chickens with MD lesions was negatively correlated with the time of death, and, similar to feather pulp, none of the surviving chickens was virus positive at the end of the experiment. Quantization of the viral genome titers in feather tip extracts at 27 and 38 dpe revealed a positive correlation with the presence of MD lesions, but only in the declining phase (38 dpe) and not at the peak (27 dpe) of the viral titer. Sire effects were significant, indicating the presence of genetic factors that affect viral proliferation. Again, significance was only observed at 38 dpe and not at 27 dpe. The results indicate that, in this commercial line, 1) all chickens were susceptible to infection via contact exposure, 2) all surviving chickens recovered from the viral infection, and 3) it is not sufficient to measure viral titers at a single time point when using viral titers as an endpoint for MD susceptibility.

Campylobacter jejuni is the leading cause of food-borne bacterial gastroenteritis in humans in the United States. Infectious bursal disease virus (IBDV) causes an immunosuppressive disease in young chickens. To analyze a possible role of IBDV-induced immunosuppression in colonization and shedding of C. jejuni, two experiments were conducted. In both experiments, group 1 consisted of noninoculated control chickens, groups 2 and 3 were inoculated with varying doses of C. jejuni, and groups 4 and 5 were inoculated initially with IBDV followed by doses of C. jejuni similar to groups 2 and 3. Campylobacter jejuni was recovered from the cloaca and cecum, but not the small intestines, from all chickens in groups 2 and 3. In groups 4 and 5, C. jejuni was recovered from the small intestines, cecum, and cloaca from all chickens. The amount (colony-forming units/sample) of C. jejuni recovered from chickens in groups 4 and 5 was significantly greater (P < 0.05) than the amount recovered from chickens in groups 2 and 3; and C. jejuni was also present sooner in these groups than in groups 2 and 3. Bursa samples from chickens in groups 4 and 5 were significantly smaller (P < 0.05) than in the other groups. Additionally, real-time polymerase chain reaction results for IBDV were positive in groups 4 and 5 and negative in all other groups. This study indicated that IBDV infection exacerbated colonization and shedding of C. jejuni, presumably through the immune suppression this virus causes in chickens. It highlights the need for further investigation into the role of immunosuppression in preharvest control strategies for food-borne disease-causing agents.

In the last 2 decades, the prevalence of Salmonella enterica serotype Enteritidis (Salmonella Enteritidis) has dramatically increased worldwide, becoming the leading cause of food-borne illnesses and an important public health issue. Many studies have suggested the role of the SEF14 fimbrial protein in the adhesion of Salmonella Enteritidis to the host. In the present study, the sefA gene, which encodes the main subunit of the SEF14 fimbrial protein, was cloned into a temperature-sensitive expression vector and transformed into a nonpathogenic, avirulent strain of Escherichia coli. The recombinant strain was used as a vaccine to elicit specific immune response against the SefA protein of Salmonella Enteritidis in 1-day-old chickens. The recombinant strain was reisolated from the intestines of treated birds for up to 21 days posttreatment, demonstrating its ability to colonize the intestinal tracts of 1-day-old chickens. In addition, immunoglobulin A (IgA) against the SefA protein was detected in intestinal secretions from treated birds at 7 days posttreatment and in bile samples from 14 to 21 days posttreatment by enzyme-linked immunosorbent assay. Nontreated birds did not show any evidence of intestinal colonization by the recombinant strain or anti-SefA IgA response in their bile or intestinal secretions. Preliminary evaluation of the recombinant strain showed a potential use of this strain to elicit protection against Salmonella Enteritidis infection in chickens. Further experiments are needed to study the ability of the recombinant strain to protect birds against Salmonella Enteritidis colonization.

In this study, a Chinese field strain of subgroup J avian leukosis virus (ALV-J), NX0101, was studied for its immunosuppressive effects in both commercial broilers and SPF white Leghorn chickens infected at 1 day of age. Our data demonstrated that NX0101 induced much more significant body and immune organ weight loss in the infected commercial broiler chickens in an earlier age than that in the SPF white Leghorn chickens. At the same time antibody responses to vaccinations of Newcastle disease virus (NDV) and infectious bursa disease virus (IBDV) in the NX0101-infected chickens were also evaluated and compared between the commercial broiler chickens and the SPF white Leghorn chickens. Compared with the control group of chickens, the hemagglutination inhibition (HI) antibody response to NDV vaccines was significantly reduced in the NX0101-infected commercial broiler chickens from as early as 20 days after vaccination. However, no significant difference in HI antibody response was seen when HI titers reached their peaks in the NX0101-inoculated and control SPF white Leghorn chickens, except it declined significantly faster in infected birds. Neither of these two types of chickens showed significant decrease of antibody response to IBDV vaccination. Herein, we conclude that this NX0101 strain of ALV-J could selectively suppress humoral immune reactions to NDV, especially in broilers. But challenge experiments were not conducted and, therefore, it cannot be known if decreased antibody levels correlated with decreased protection against NDV in this case.

The purpose of the present cross-sectional study was to evaluate the health status of organic broiler chickens and the contamination rate with Salmonella and Campylobacter in organic broiler production in Belgium. The broilers were screened for antibodies against routinely monitored poultry diseases at 1 day old and at slaughter. Fecal examination for the presence of worm eggs was done at slaughter. Bacteriological examination for the detection of Salmonella and Campylobacter was performed at day 1, week 2, week 4, week 7, week 10, and slaughter. Conventional broilers of the same poultry integration and reared in the same geographic area were also screened and served as reference. Serologic data indicated lower antibody titers against infectious bronchitis and Newcastle disease in organic flocks. No significant differences could be found in prevalence of Salmonella between organic and conventional broilers at slaughter. In contrast, Campylobacter infections at slaughter were significantly higher in organic flocks. Organic flocks most probably become infected with Campylobacter between week 7 and week 10. Worm eggs were found in neither the organic flocks nor the conventional flocks. In conclusion, there are indications that the respiratory health status is better in organic broilers but that organic flocks are more often infected with Campylobacter than are conventional flocks.

An outbreak of subcutaneous sarcomas in commercial White Leghorn egg layers was observed in the northeastern United States during late 2004. Subcutaneous tumors were confined to three flocks distributed in two locations and belonging to the same company. The tumors were first observed grossly by farm personnel at approximately 7 wk of age and persisted throughout the economic life of the flocks. Most of the tumors observed during the growing period were present on the facial region or around the head, wings, and legs. There was no gross evidence of bursal or visceral involvement. Microscopically, most tumors were undifferentiated sarcomas and myxomas. There was no microscopic evidence of Marek's disease or lymphoid leukosis. Reticuloendotheliosis virus proviral DNA was not detected by polymerase chain reaction either in tumors or in cell cultures. Egg production and mortality rates were within normal limits in the affected flocks and many of the chickens exhibiting tumors seemed healthy otherwise, albeit approximately one-half of the daily mortality exhibited tumors. Avian myeloblastosis-associated virus type 1 (MAV-1) was isolated from tumors, plasma, and serum. Upon initial virus neutralization, the viruses isolated seemed at least partially related antigenically to avian leukosis virus (ALV) subgroups A and B but not to subgroup J (ALV-J). Sequencing of the variable and hypervariable regions of gp85 in the envelope gene revealed that the viruses involved are closely related to MAV-1. Attempts to reproduce subcutaneous sarcomas with MAV-1 isolated from White Leghorn chickens in the case herein reported produced exclusively myelocytomas indistinguishable histologically from those induced by ALV-J in meat type chickens.

Avian leukosis virus (ALV) infection in chickens is known to induce increased mortality, tumors, delayed growth, and suboptimal egg production. Countries importing specified pathogen-free eggs, vaccines, and poultry breeding stock require freedom of infection or contamination with ALV in such products among other avian pathogens. Recently, ALV was found as a contaminant in a limited number of commercial poultry vaccines, even after routine quality assurance procedures cleared the vaccines for commercialization. The contaminated vaccines were promptly withdrawn from the market, and no direct detrimental effects were reported in poultry vaccinated with such vaccines. We describe herein the characterization in vitro of the contaminant viruses. All exogenous viruses detected in four vaccine lots belong to subgroup A of ALV based on cell receptor interaction, subgroup-specific polymerase chain reaction (PCR), envelope gene sequencing, and virus neutralization. A combination of thermal treatment and serial dilutions of the contaminated vaccines facilitated detection of contaminating ALVs in cell culture coupled with antigen-capture enzyme-linked immunosorbent assay. Subgroup-specific PCR readily detected ALV-A directly in the contaminated vaccines but not in naive vaccines or cell controls. Our methods are proposed as complementary procedures to the currently required complement fixation for avian leukosis test for detection of ALV in commercial poultry vaccines.

Recombinant avian adeno-associated viruses coding for the LacZ gene were used to inoculate embryonating chicken eggs, to assess the usefulness of the system for the expression of a transgene in vivo. The results obtained indicate significantly higher levels of expression of the reporter gene at various time intervals in the embryos inoculated with the recombinant virus in comparison with the mock-inoculated controls. At the embryo level, significant differences were evident at 120 hr postinoculation; hatched chicks showed transgene expression up to 14 days of age. In a second experiment, different cell-line cultures were transfected with plasmids encoding for a reporter gene flanked by the avian adeno-associated virus inverted terminal repeats (ITR), either alone or in the presence of the major nonstructural proteins of the virus (Rep 78/68) to assess the ability of these proteins and DNA elements to enhance gene expression. Results indicate that the inclusion of the viral ITR alone or during coexpression of the Rep proteins significantly enhances the expression of the transgene in all cell lines tested, as evidenced by the detection of the β-galactosidase protein through chemiluminescence reactions and staining of transfected monolayers.

Eight poultry farms in Nigeria, including chickens from nine breeder, 14 broiler, 28 pullet, 11 layer, and three cockerel flocks, were tested for antibody seroprevalence to the following poultry viruses of potential economic importance: infectious bronchitis virus (IBV), avian reovirus, avian pneumovirus (APV), infectious laryngotracheitis virus (ILTV), avian influenza virus (AIV), and avian leukosis virus (ALV). Serum samples were collected between 1999 and 2004 and were tested for antibodies using commercial enzyme-linked immunosorbent assay (ELISA) kits. Seroprevalence was very high for IBV (84%); intermediate for reovirus (41%), APV (40%), and ILTV (20%); and very low for ALV (<5%) antibodies. By commercial ELISA, the seroprevalence of antibodies against AIV was, in some flocks, up to 63%. However, more specific assays did not confirm AIV antibodies, indicating that all flocks tested were free of avian influenza antibodies. Birds seemed to be first infected by IBV (at about 7 wk of age), then by reovirus at 12 wk, before they became infected by APV (week 25) and ILTV (week 30). This is the first report of serological evidence of the above viruses in West Africa. Further studies are necessary to assess economic losses due to these avian viruses and the costs and benefits of countermeasures.

The duration of protective immunity elicited by the MS-H vaccine was evaluated by experimental challenge of chickens at 15 and 40 wk after eyedrop vaccination. Immunity induced by the parent strain of the vaccine, 86079/7NS, was also investigated for comparison. A serological response to Mycoplasma synoviae was detected in 89% to 100% of MS-H vaccinates and 86079/7NS inoculates at 15, 27, 30, 35, and 40 wk after inoculation. A significantly lower incidence of air-sac lesions and lower air-sac lesion severity were observed in both the MS-H vaccinated and the 86079/7NS inoculated groups, as compared to the unvaccinated controls, after both challenge points. Tracheal mucosal thicknesses in MS-H vaccinates was significantly lower in the upper, lower, and total trachea at 40 wk after vaccination, as compared to the controls. It was demonstrated in this experiment that protective immunity, as determined by protection against experimental challenge, was maintained to at least 40 wk after vaccination.

Recently, avian leukosis virus (ALV) was isolated from four lots of Marek's disease vaccine produced by two laboratories. The ALVs isolated were characterized by examination of their interactions with cells of two phenotypes (C/E and C/A,E), subgroup-specific polymerase chain reaction (PCR), virus neutralization, envelope gene sequencing, and phylogenetic analysis. All four ALVs are exogenous, belong to subgroup A, and appear to be virtually identical to each other based on PCR and envelope gene nucleotide sequences. We describe herein the characterization of the contaminant viruses in vivo by means of experimental infection in chickens. The contaminant viruses established transient viremia in specified pathogen-free (SPF) Leghorn chickens and elicited a robust and lasting antibody response detectable by enzyme-linked immunosorbent assay. None of the contaminant ALVs induced tumors up to 31 wk of age, and mortality was insignificant. Despite a strong antibody response against the contaminant ALVs, vertical (congenital) transmission to the progeny of experimentally infected SPF chickens took place, albeit at a very low rate (≤1.6%). Experimental infection in meat-type chicken embryos resulted in viremia at hatch, suggesting that some meat-type chickens are susceptible to infection and support virus replication.

The objective of this study was to examine the feasibility of using aerosolized fluorescent microspheres to examine particle distribution in the respiratory tract of birds following aerosol exposure. Adult domestic pigeons (Columbia livia domestica; n = 5 birds per microsphere size) were exposed to aerosolized monodispersed populations of various sized carboxylate microspheres (0.5, 1.0, 2.0, 3.0, 6.0, and 10.0 μm) for 30 min. For aerosol-exposure purposes, the birds were anesthetized with injectable anesthetics, intubated, and placed on positive-pressure ventilation using a mechanical ventilator. Immediately following aerosol exposure, the birds were euthanatized, and carcasses were preserved via intravenous infusion of modified paraformaldehyde/gluteraldehyde fixative (pH = 7.2 and 340 mOsm). Initial evaluation of microsphere distribution in air sacs (cranial and caudal thoracic and abdominal) and at the level of the ostia was performed using a stereoscopic microscope with an epifluorescent module. More detailed examination of the distribution of microspheres within the respiratory tract was achieved using a confocal scanning laser microscope with a krypton argon laser and a scanning electron microscope. The results from this study revealed that positive-pressure ventilation resulted in distribution of smaller sized fluorescent microspheres (sizes 1.0, 2.0, and 3.0 μm) throughout the pigeon's respiratory tracts, and these microspheres were in highest concentration in the secondary bronchi and ostia for all of the examined air sacs. The larger sized beads (6.0 and 10.0) were confined to the upper airway (trachea and primary bronchi). The results from this study allow for a better understanding of particle deposition following positive-pressure ventilation and aerosol exposure in birds.

Infectious bursal disease (IBD) viruses detected in commercial flocks of different regions of Argentina were analyzed by reverse transcription–polymerase chain reaction–restriction fragment length polymorphism (RFLP) of a VP2 gene fragment, followed by sequence analysis. Two out of eight IBD viruses presented an SspI restriction site, typical of the very virulent phenotype. Three IBD viruses presented a SacI restriction site, typical of classic virulent strains, and one isolate presented restriction sites for both enzymes. The Argentine IBD viruses showed three different molecular patterns by RFLP with the restriction endonuclease BstNI and five different patterns with MboI. By comparison of nucleotide and deduced amino acid sequences of the hypervariable region of the VP2 protein, four Argentine viruses were found to be closely related to Brazilian subclinical strains and two isolates were found to be related to vaccine IBDV strains in use in Argentina. Strain LD9569 was genetically characterized as a very virulent strain and was found to be closely related to international and regional vvIBDV strains. This is the first report on variability of IBDV strains circulating in Argentina.

Eastern Screech Owls (EASOs) were experimentally infected with the pathogenic New York 1999 strain of West Nile virus (WNV) by subcutaneous injection or per os. Two of nine subcutaneously inoculated birds died or were euthanatized on 8 or 9 days postinfection (DPI) after <24 hr of lethargy and recumbency. All subcutaneously inoculated birds developed levels of viremia that are likely infectious to mosquitoes, with peak viremia levels ranging from 10^5.0 to 10^9.6 plaque-forming units/ml. Despite the viremia, the remaining seven birds did not display signs of illness. All birds alive beyond 5 DPI seroconverted, although the morbid birds demonstrated significantly lower antibody titers than the clinically normal birds. Cagemates of infected birds did not become infected. One of five orally exposed EASOs became viremic and seroconverted, whereas WNV infection in the remaining four birds was not evident. All infected birds shed virus via the oral and cloacal route. Early during infection, WNV targeted skin, spleen, esophagus, and skeletal muscle. The two morbid owls had myocardial and skeletal muscle necrosis and mild encephalitis and nephritis, whereas some of the clinically healthy birds that were sacrificed on 14 DPI had myocardial arteritis and renal phlebitis. WNV is a significant pathogen of EASOs, causing pathologic lesions with varying clinical outcomes.

Recombinant viruses were rescued after site-specific mutagenesis of a full-length clone of the lentogenic Newcastle disease virus (NDV) strain Clone 30. To assess the contribution of different amino acids to virulence, specific alterations were introduced into the fusion (F) protein and in the hemagglutinin-neuraminidase (HN) protein based on sequence comparison between NDV strains of different virulence. Modification of the proteolytic cleavage site in the F protein to a polybasic motif increased the intracerebral pathogenicity index (ICPI) from 0.0 to 1.28. Moreover, the additional exchange of amino acid 123 of the HN protein from tryptophan to cysteine in combination with alteration of amino acid 27 of the F protein from cysteine to arginine increased the ICPI to 1.5. The HN mutation visibly altered conformation of the protein, resulting in the formation of disulfide-linked HN dimers that may indicate that this HN conformation is beneficial for the virulent phenotype.

A latex agglutination test (LAT) was developed for rapid detection of antibodies to H5 avian influenza viruses (AIVs). The hemagglutinin protein of H5 AIV was covalently linked to carboxylated latex by ethyl-dimethyl-amino-propyl carbodiimide to prepare the sensitized latex beads. The LAT was evaluated with the hemagglutination inhibition (HI) assay as the reference test. The H5-LAT showed a sensitivity of 87.0% and specificity of 88.9% in detecting 126 serum samples from experimentally infected chickens and a sensitivity of 82.5% and specificity of 86% in detecting 587 field chicken serum samples from mostly vaccinated chickens. The agreement ratio between H5-LAT and HI was found to be 87.3% and 83.1% for the two groups of samples, respectively. Difficulty with background agglutination in stored chicken sera was overcome by serum pretreatment with either dried chicken liver powder or dilution buffer containing detergent Tween-20. The H5-LAT has advantages over a previously reported whole-virus LAT in terms of biosafety in preparation, chemical stability, and higher specificity. It is a rapid and simple test suitable for field monitoring of antibodies to H5-type AIV.

Resistance to infection with avian influenza virus (AIV) was studied in healthy and immune-suppressed pigeons, which were treated with the immunosuppressant cyclophosphamide (Cy) before infection. Two subtypes of low pathogenic AIV (LPAIV; CK/TW/H5 and CK/TW/H6) were inoculated via the oculonasal route. Nested reverse transcriptase-polymerase chain reaction (nested RT-PCR) and virus isolation were used as detection methods. The Cy-treated and -untreated pigeons in both experiments did not shed viruses or become antibody positive throughout the 21-day observation period. All pigeons were negative for AIV RNA when trachea, lung, pancreas, spleen, kidney, and rectum tissues were examined. Negative results were also obtained in uninoculated contact chickens, which were housed together with H6N1 AIV-inoculated pigeons. Therefore, it was concluded that the pigeons are resistant to infection with these two LPAIVs and do not serve as transmission hosts, even in the presence of immune dysfunction.

Colibacillosis accounts for significant losses to the poultry industry, and control efforts are hampered by limited understanding of the mechanisms used by avian pathogenic Escherichia coli (APEC) to cause disease. We have found that the presence of the increased serum survival gene (iss) is strongly associated with APEC but not with commensal E. coli, making iss, and the protein it encodes (Iss), candidate targets of colibacillosis control procedures. To assess the potential of Iss to elicit a protective response in chickens against APEC challenge, Iss fusion proteins were produced and administered subcutaneously to four groups of 2-wk-old specific-pathogen-free leghorn chickens. At 4 wk postimmunization, birds were challenged with APEC from serogroups O2 and O78 via intramuscular injection. At 2 wk postchallenge, birds were necropsied, and lesions consistent with colibacillosis were scored. Also, sera were collected from the birds pre- and postimmunization, and antibody titers to Iss were determined. Immunized birds produced a humoral response to Iss, and they had significantly lower lesion scores than the unimmunized control birds following challenge with both APEC strains. Birds that received the smallest amount of immunogen had the lowest lesion scores. Although further study will be needed to confirm the value of Iss as an immunoprotective antigen, these preliminary data suggest that Iss may have the potential to elicit significant protection in birds against heterologous E. coli challenge.

The progress and transmission of blackhead disease in chickens was studied in battery cages and floor pens in the absence of vectors. Two-week-old chicks were inoculated intracloacally with Histomonas meleagridis and allowed to commingle with others in floor pens. There was no confirmed transmission of blackhead to other birds in the pen, whether stocked at 10% or 25% with infected birds. A second experiment evaluated the effects of feed restriction of chickens on spread of blackhead within floor pens. Inoculated seeder birds had severe cecal lesions of blackhead at necropsy, regardless of feed restriction. Uninoculated birds did not develop lesions by the time of necropsy at 42 days of age, regardless of whether full-fed or limited by skip-day feeding. Chickens inoculated intracloacally with H. meleagridis and placed in battery cages became infected and had cecal lesions of blackhead, but few liver lesions. Chickens allowed to commingle with the inoculated birds in batteries had no lesions of histomoniasis at necropsy 2 wk postinoculation. Coccidial oocysts from turkeys (Eimeria adenoeides) were inoculated along with H. meleagridis from cultures to test the effects of sporozoite penetration in the ceca on progress of blackhead disease. Histomoniasis was not worsened by the interaction with sporozoites, as shown by unchanged severity of cecal lesions, the number of birds showing liver lesions, or the overall number of positive birds. Overall, blackhead infections showed no inclination to spread from bird to bird under conditions of these studies, in contrast to what has been reported for turkeys. These results suggest that the dynamics of blackhead transmission in chickens differs significantly from that of turkeys, where transmission from bird to bird is rapid and effective in the absence of vectors.

Fowl typhoid is under control in poultry farms of developed countries, but it still endemically subsists in commercial laying hen farms of some countries. It has been demonstrated that Salmonella live vaccines can elicit cross-immunity against members of the same Kauffmann–White scheme serogroup. In this work, we explored the protection conferred by TAD Salmonella vac^® E, a live Salmonella enterica serovar Enteritidis vaccine, against fowl typhoid. Three groups of laying hens were vaccinated with different vaccination schedules starting on the first day of life, and afterwards were infected with 2 × 10⁵ CFU of a virulent Salmonella Gallinarum strain, either at wk 28 or wk 52. Mortality, fecal shedding, and organ invasion of Salmonella Gallinarum were assessed. In this work we demonstrated that this Salmonella Enteritidis vaccine is able to cross-immunize against Salmonella Gallinarum. At wk 28, hens vaccinated with three oral doses or with two oral doses combined with one subcutaneous dose were protected by the vaccine. At wk 52, when hens were infected 36 wk after the final immunization, the vaccine was not able to confer protection. Thus, revaccination every 3 mo would be highly recommended. In countries where Salmonella Gallinarum subsists together with Salmonella Enteritidis, control programs should include vaccination of laying hens using safe attenuated Salmonella strains.

The transmission of pathogens from infected to susceptible hosts may occur through contaminated fomites and inanimate objects. This type of transmission depends on the ability of the pathogens to survive in the environment. In this report, we describe the survivability of two avian respiratory viruses, e.g., avian metapneumovirus and avian influenza virus on 12 different porous and nonporous surfaces. The viruses survived on some of the surfaces for up to 6 days postcontamination but not after 9 days. Both viruses survived longer on nonporous surfaces than on porous ones. One of the reasons for poor survival on porous surfaces could be inefficient elution of virus from these surfaces. These results should be helpful in determining how long the premises should be left vacant after an outbreak of these viruses has occurred in poultry houses.

Pathogenicity of two avian poxviruses isolated from endangered Hawaiian wild birds, the Hawaiian Goose and the Palila, was compared with fowl poxvirus in chickens. Immune responses were measured by ELISA pre- and postimmunization with Hawaiian poxviruses and after challenge with fowl poxvirus. Both isolates from Hawaiian birds developed only a localized lesion of short duration at the site of inoculation in specific-pathogen-free chickens and did not provide protection against subsequent challenge with virulent fowl poxvirus. On the other hand, birds inoculated with virulent fowl poxvirus developed severe lesions. In contrast to high antibody response in chickens immunized with fowl poxvirus, birds immunized with either of the two Hawaiian isolates developed low to moderate antibody responses against viral antigens. The level of immune responses, however, increased in birds of all groups following subsequent challenge.

The objective of this study was to compare the presence of the Arkansas (Ark) and Massachusetts (Mass) serotypes of infectious bronchitis virus (IBV) in the tracheas and cecal tonsils of commercial broilers after vaccination at 1 day of age by coarse spray. When given as a single serotype vaccine, the Mass strain was detected by reverse transcriptase–polymerase chain reaction (RT-PCR)–restriction fragment length polymorphism (RFLP) only in the tracheas, whereas the Ark strain was detected in both the tracheas and cecal tonsils. By in situ hybridization, the Mass and Ark nucleocapsid (Nc) genes were detected only at 7 days in the tracheas. When both strains were given in the mixed vaccine, the Mass strain was more consistently detected by RT-PCR–RFLP in the tracheas and cecal tonsils at early stages of infection (up to 14 days) and the Arkansas strain was more consistently detected at late stages of infection (21 and 28 days). By in situ hybridization, the IBV Nc gene was more consistently detected in the trachea at early stages of infection (7, 14, and 21 days) and in the cecal tonsils at late stages of infection (21, 28, and 35 days). In general, the Mass strain was more frequently recovered from the tracheal and cecal tonsil tissues at earlier stages of infection and the Ark strain was recovered at later stages of infection.

The ileal Peyer's patches (Pp), secondary gut-associated lymphoid tissue of the mucosal immune system, may serve as an important site for monitoring inflammatory and immunologic responses of the host against enteric pathogens. Chicken Pp are often difficult to observe grossly, and a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a novel staining method that is quick, easy, and accurate to aid in gross identification and recovery of the chicken Pp from fresh tissue specimens. Lower alimentary tracts were harvested from White Leghorn hens and commercial broilers. The ileocecocolic region was excised intact, flushed with deionized water to remove ingesta, and a dilute eosin-Y solution was infused. After 1 min, the eosin-Y was gently extruded. Modified-crystal violet (mCV) was then injected into the gastrointestinal segment, where on the lymphoid tissue area became apparent at the serosal surface. The distal ileal Pp was visible as a pale whitish pink ovoid-focalized area with surrounding gut tissue stained light purple. The exact Pp site could be delineated at the serosal and mucosal surface by gross assessment. Light microscopy evaluation of hematoxylin and eosin-stained tissue slides prepared from the excised Pp site revealed lymphoid tissue aggregations with multiple follicular units indicative of Pp. The novel eosin-Y mCV staining technique promotes rapid identification and accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens.

The effect of a systemic disease on the dynamics of iron, zinc, and copper in chickens fed ad libitum was examined by infecting 10-day-old specific pathogen-free chickens with infectious bursal disease virus (IBDV). Liver, bursa of Fabricius, pancreas, spleen, and serum were sampled in 10 controls and 10 challenged chickens at 3-day intervals postinfection (PI) for 15 days. The samples were analyzed using atomic absorption spectroscopy. Serum levels were similar to that reported in the literature. Concentrations of iron and zinc did not change significantly in the pancreas, but there was an increase in copper in infected pancreatic tissue on days 9 and 15 PI. Iron concentration in the spleen showed a significant increase on days 6, 9, and 15 PI, whereas zinc was only significantly increased on day 15 PI. There was no significant change in copper concentrations in the spleens of infected chickens vs. controls. This finding is in line with previously reported data.

The results showed that the liver was not a major tissue where iron and zinc were sequestered, as previous data have shown in mammals. Instead, the bursa of Fabricius had significantly increased levels of both iron and zinc in infected tissue vs. control tissue from 9 days PI on. Furthermore, the bursa had increased levels of copper in the latter portion of the study. These findings suggest that the bursa of Fabricius rather than the liver is the major organ for metallic ion sequestering during IBDV infection.

Erysipelas was diagnosed in a layer breeder flock in Sweden in 2002. Although vertical transmission of Erysipelothrix rhusiopathiae has not been previously described in chickens, the potential of erysipelas infection to adversely affect hatching eggs was of concern. To clarify the possible impact of erysipelas on hatching eggs and their progeny, an experiment was done using 200 hatching eggs collected from the infected flock. The eggs were incubated for 21 days, and the egg shells, infertile eggs, dead-in-shell embryos, and a sample of day-old hatched chicks and blood samples from 5-day-old chicks were cultured for E. rhusiopathiae. In addition, after 28 days of grow-out, the male chickens were euthanatized and cultured for the bacterium, and the remaining female chickens were placed as a backyard flock and observed over a 4-mo period. Bacteriological test results of the above-mentioned samples were negative for E. rhusiopathiae. Mortality rates were not excessive, and no clinical symptoms of erysipelas were observed during the period of observation. The result of the investigation suggests that in layer breeder chickens, E. rhusiopathiae is not vertically or egg transmitted and that the disease outbreak in the parent stock had no adverse impact on the quality of hatching eggs in terms of increased embryo mortality.

Increased trimming and consequent carcass downgrade was experienced in a broiler flock. Condemned birds had gray to black pigmentation of the subcutaneous tissue and fat of the abdomen, and bluish pigmentation of the shanks. Histopathology revealed accumulation of melanin in these tissues. No significant bacteria were isolated. Accumulation of melanin in the skin and fascia is because of the cumulative and interactive effects of several genes. Additionally, pigmentation of abdominal fat has a high correlation with accumulation of melanin in the skin of the shanks. Although melanization of the skin and abdominal fascia is not harmful to people, it may cause severe economic losses to the producer.