Early events leading to the establishment of various branches of the U.S. poultry industry and the animal health industry in the United States are reviewed. Entrepreneurs see needs in a developing market and take action to meet these needs. Their skills are suited to organize people and processes to supply these needs. As the early poultry industry consolidated, this was followed by technical education as a marketing tool to expand access to the customer base. Focus will be on two entrepreneurial individuals not well covered by prior history articles, namely Hiram Lasher and Kenneth Eskelund. A discussion of how industries pass from an entrepreneurial phase to a corporate phase will follow, thus explaining the corporate- and biotechnology-oriented vaccine companies we see today.

Avian colibacillosis resulting from avian pathogenic Escherichia coli (APEC) seriously disrupts poultry production. Hatcheries are the main source of chickens for commercial farms. To characterize the potential pathogenicity of E. coli strains isolated from hatcheries, 2344 fluff samples from 1-day-old chickens were collected from hatching incubators between October 2016 and November 2017. Among the hatcheries, the incidence of E. coli varied from 0% to 16.9%, with an overall incidence of 2.0%. High incidences reflected inadequate sanitation in some hatcheries. We also compared 20 clinically isolated APEC strains with fluff-originated E. coli in terms of existence of 10 virulence-associated genes (VAGs) and antimicrobial-resistance genes, and antimicrobial resistance using minimum inhibitory concentration (MIC) values. Our results showed that APEC more-frequently possessed most of the assessed VAGs (papC, astA, cvaC, hlyF, fyuA, iroN, iutA, iss, and ompT), suggesting that fluff-originated E. coli is less likely to cause avian colibacillosis. However, fluff-originated E. coli more-frequently expressed the adhesion gene fimC, which could confer higher upper respiratory tract adhesion. Both APEC and fluff-originated E. coli demonstrated multidrug resistance including 100% resistance to ampicillin, amoxicillin, cephalexin, florfenicol, and trimethoprim-sulfamethoxazole. Based on median MIC values, fluff-originated E. coli was more susceptible to antibiotics. However, resistance-gene existence did not significantly differ between groups, suggesting that fluff-originated E. coli should still be a public health concern. Molecular subtyping with XbaI-digested pulsed-field gel electrophoresis revealed that only a few strains showed identical patterns, indicating that a variety of contamination sources were present within individual hatcheries. Identical strains within the same hatchery may indicate vertical transmission from parent flocks. Overall, this is the first study to characterize fluff-originated E. coli. Our results suggest that it has lower pathogenicity than APEC and that thorough sanitation should be performed to reduce the occurrence of fluff-originated E. coli in hatcheries.

We have examined a variety of sampling strategies for detecting pathogens in turkey flocks undergoing infections with low pathogenicity avian influenza virus (LPAIV). We found that viral RNA was widely distributed in the barn environment of turkey flocks undergoing an active LPAIV infection and was in both water and drinker biofilm samples. Viral RNA was concentrated in drinker biofilm and sediment and was detectable using real-time reverse-transcription polymerase chain reaction (RRT-PCR) and by virus isolation. Drinker biofilm sample results correlated with concurrently collected oropharyngeal (OP) sample results from flocks on a farm with LPAI in which the two sampling strategies were directly compared. To evaluate the utility of biofilm sampling for the detection of highly pathogenic avian influenza virus (HPAIV), biofilm and OP swabs from mortality pools were collected daily from negative turkey flocks on an HPAI-positive premise. The biofilm swabs were positive 1–2 days prior to positives appearing in the OP sample pools. The drinker biofilm sampling strategy overcame the difficulty of finding a subclinical infectious bird in a population by collecting material from a large number of individuals and testing a sample in which a positive signal persists for several days to weeks. The sampling method is convenient for use in turkey barns and has been reliably used in both active and passive surveillance programs for LPAIV and HPAIV using RRT-PCR.

In Morocco in early 2016, a low pathogenic avian influenza virus serotype H9N2 caused large economic losses to the poultry industry, with specific clinical symptoms and high mortality rates on infected farms. Subsequent to the H9N2 outbreak, the causal agent was successfully isolated from chicken flocks with high morbidity and mortality rates, propagated on embryonated eggs, and fully sequenced. The phylogenetic analysis suggested that the Moroccan isolate could have derived from the Middle East isolate A/chicken/Dubai/D2506.A/2015. This study was designed to assess the pathogenicity of the Moroccan isolate H9N2 in experimentally infected broiler and specific-pathogen-free (SPF) chickens. At 22 days of age, one broiler and two SPF chicken groups were inoculated by dropping 0.2 ml of the H9N2 isolate (10^7.5 EID₅₀/ml) in both nostrils and eyes. Clinically inoculated chickens with H9N2 displayed mild lesions, low mortality rates, and an absence of clinical signs. The H9N2 virus was more pathogenic in broiler chickens and produced more severe tissue lesions compared to SPF chickens. The viral shedding was detected up to 6 days postinoculation (pi) in oropharyngeal and cloacal swabs in infected birds with a maximum shedding in the oropharynges of the broiler group. All experimental chickens seroconverted and registered high hemagglutination inhibition titers as early as day 7 pi. The present study indicates that the H9N2 virus isolated from a natural outbreak was of low pathogenicity under experimental conditions. However, under field conditions infection with other pathogens might have aggravated the disease.

Chlamydia psittaci is a zoonotic pathogen with multiple hosts, especially avian, and can be transmitted to humans, causing psittacosis or ornithosis. No effective vaccines have been developed. We therefore isolate and genotype avian C. psittaci strains and investigate the pathogenicity of isolates in the southern Hunan area of China. Among 200 suspicious avian specimens, eight were positive for the C. psittaci outer membrane protein A (ompA) gene (4%), and seven were successfully cultured in human epithelial type 2 and Vero cells (87.5%). Genotyping of the ompA gene of the eight PCR-positive samples revealed that all of the cultured strains, except for the E9 strain, belonged to genotype A. Pathologic changes in the mice infected with C. psittaci via intranasal inoculation showed severe pneumonia and intense infiltration of inflammatory cells in the lung in a dose-dependent manner, and immunohistochemical staining displayed different levels of infiltration of C. psittaci inclusions in the heart, liver, spleen, kidney, and, especially, lung. Our findings demonstrate that genotype A dominates all C. psittaci genotypes in the southern Hunan area and that the C. psittaci avian isolates in this region possess dose-dependent pathogenicity.

Infectious bronchitis virus (IBV) is highly prevalent in broiler chickens despite extensive vaccination commonly conducted early after hatch. The effects of early vaccination on immune responses were further investigated in chickens primed at increasing ages, followed by booster vaccination with an attenuated Arkansas (Ark) Delmarva Poultry Industry–type vaccine. Results show that vaccination on day 1 of age elicits significantly lower systemic and mucosal antibody responses compared with vaccination at later time points in the life of the chicken. The increase of IBV antibodies in serum from secondary responses after booster vaccination was more dramatic and significantly higher when measured by an Ark spike subunit 1 protein ELISA compared with measuring by non-Ark serotype whole-virus ELISA, which underlines the immunogenic importance of the virus spike at inducing antibodies. However, the levels achieved following boosting did not differ significantly between ages of priming. Thus, it seems that the booster vaccination mitigated the differences detected after prime immunization. In contrast to the continued rise of systemic antibodies after booster vaccination, the levels of mucosal IBV-specific immunoglobulin A decreased after booster vaccination. The recruitment or expansion of cluster of differentiation (CD)4⁺, CD8⁺, and CD4⁺/CD8⁺ T-cell populations in different immune effector sites was increased with age, but remained unaltered by IBV vaccination. In contrast, peripheral blood CD4⁺ cells showed a significant increase in IBV-vaccinated chickens compared with nonvaccinated age-matched controls both after primary and booster immunization. The results of the current study confirm that IBV vaccination on the day of hatch induces suboptimal IBV immune responses both in the systemic and mucosal compartments. This routine practice may be contributing to the immunologic escape of the virus and increased persistence of vaccine virus in vaccinated chickens. However, booster vaccination seems to overcome poor initial responses.

The wooden breast myopathy is identified by the palpation of a rigid pectoralis major muscle and results in myofiber necrosis and fibrosis in fast-growing, meat-type broilers. The fibrosis in wooden breast–affected muscle is characterized by the replacement of myofibers with extracellular matrix proteins, especially fibril-forming collagens. Studies have shown differences in collagen organization in fast-growing broiler lines, with tightly packed and highly aligned collagen organizations having a higher phenotypic incidence of wooden breast. The objective of the current study was to analyze collagen fibril organization further in two fast-growing broiler lines (Lines A and B) with incidence of wooden breast compared with a slower growing broiler Line C with no phenotypically detectable wooden breast. The small leucine-rich proteoglycan decorin was also studied for its interaction with collagen by immunogold detection. Decorin binds to fibrillar collagens and organizes their alignment and crosslinking, both of which will affect collagen functional properties. Key findings from the study showed that collagen shifts to larger diameter collagen fibril bundles with the wooden breast myopathy. Specifically, broilers affected with wooden breast from Line A had a more dramatic shift toward larger collagen fibril bundles compared with those affected from Line B. Wooden breast–affected Line A had collagen fibril bundles up to 8.4 µm, whereas Line B maximum size was 5.1 µm. Although decorin-collagen binding was not different overall in the wooden breast myopathy or broiler line, for small-diameter collagen fibril bundles, wooden breast–affected Line A had more decorin-collagen binding than wooden breast–affected Line B. Taken together, these data provide further evidence that multiple fibrotic myopathies are likely in fast-growing meat-type broilers.

In the fifth wave of the H7N9 avian influenza epidemic, highly pathogenic avian influenza (HPAI) A (H7N9) viruses have emerged and pose a great challenge to public health and the poultry industry. In addition, there are apparent genetic and antigenic variations between the classical H7N9 avian influenza virus and the newly-emerged H7N9 virus. Therefore, an antigenic-match vaccine is required for the prevention and control of H7N9 avian influenza in poultry in China. In this study, a recombinant Newcastle disease virus (NDV)-vectored vaccine expressing the HA derived from a prevailing HPAI H7N9 virus (GD15) was generated using reverse genetics. The recombinant virus (rAI4HA) showed virus yield and growth capacity in chicken embryos comparable to the parental virus (rAI4). Expression of the HA protein was detected in chicken embryo fibroblasts inoculated with rAI4HA. A chicken immunization study demonstrated that both rAI4HA and rAI4 induced similar anti-NDV hemagglutination inhibition (HI) antibody titers at weeks 2, 3, and 4 after a single immunization. However, rAI4HA-immunized chickens had a low seroconversion rate (20%) and negative HI titers against H7N9. Additionally, rAI4HA elicited high levels of H7N9-specifc IgY antibody as measured by ELISA. More importantly, the recombinant vaccine provided a complete protection against a lethal challenge with HPAI H7N9 virus and significantly inhibited virus shedding after a single immunization. Our results suggest that the recombinant NDV-vectored H7N9 vaccine expressing the antigenic-match HA can confer a complete protection against HPAI H7N9 challenge after a single immunization.

During 2015, duck farms (n = 27) in Sharkia Province, Egypt, experienced several disease outbreaks leading to mortality and nervous manifestations. Upon necropsy, the affected ducklings showed liver lesions, such as hemorrhage or necrosis, suggestive of duck virus hepatitis (DVH). Reverse transcription-PCR (RT-PCR), on the basis of the 3D gene, found duck livers from 21 farms to be positive for duck hepatitis A virus serotype 1 (DHAV-1). All duck breeds (Pekin, Mallard, and Muscovy) were infected. The virus was isolated in embryonated chicken eggs, which showed embryonic mortality (40%–80%) within 5–7 days, stunting or dwarfing (69.6%), and necrotic liver foci (60.9%). The VP1 gene of 11 DHAV-1 strains was characterized by RT-PCR and Sanger sequencing. All study strains were clustered in a monophyletic branch within subclade B2 of Group 4 and were separated from the Egyptian vaccine strain. Several amino acid (aa) residues, such as V129, S142 (only in four strains), L181, G184, and K217, were related to virus attenuation. However, two aa residues (N193 and E205), found in virulent DHAV-1 strains, were also observed in our strains. This study confirms the circulation of DHAV-1 (subclade B2) in Lower Egypt and elucidates the phylogenetic characters of the VP1 genes, which will be useful in following the local trends of DHAV-1 infections. Further studies are indicated to determine the correlation between these mutations and the virulence of the Egyptian DHAV-1 isolates.

Turkey herpesvirus (HVT) is widely used as a vaccine against Marek's disease in chickens and recently as a vector for foreign genes from infectious bursal disease virus, Newcastle disease (ND) virus, infectious laryngotracheitis (ILT) virus, and avian influenza virus. Advantages of HVT-vector vaccines are that the vaccines do not contain live respiratory viruses or live infectious bursal disease virus able to replicate and cause disease or embryo mortality, they can be administered at hatch or in ovo, and they are relatively insensitive to interference from maternally derived antibodies. As producers have tried to combine HVT-vector vaccines to protect against additional diseases, reports have indicated that applying two vectored vaccines using the same HVT vector is reported to reduce the efficacy of one or both vaccines. To confirm this interference, we evaluated commercial vaccines from multiple companies, including products with inserts designed to protect against ND, infectious ILT, and infectious bursal disease (IBD). Using a standard dosage, we found that the ILT product was most severely affected by the addition of other vaccines, as demonstrated by a significant increase in clinical signs, significant decrease in weight gain, and increase in quantity of challenge virus observed from tracheal swabs collected from Days 3–5 postchallenge. The ND and IBD products were also affected by the addition of other vaccines, although in most cases differences compared to vaccination with the vector alone were not statistically significant. This study demonstrates the importance of following manufacturer guidelines and the need for validating alternative strategies to benefit from the high level of protection offered by vector vaccines.

This study aimed to investigate the feasibility of propagating and titrating hemorrhagic enteritis virus (HEV) in chicken embryos. A total of 308 embryonated eggs were used. At 10 days of embryonic age, eggs were inoculated via allantoic sac or chorioallantoic membrane routes with non–heat-treated (live) HEV or heat-treated (dead) HEV or served as negative controls. Allantoic fluid retrieved at 0, 1, 3, 5, and 7 days postinoculation (dpi) was tested for HEV by quantitative PCR. Inoculation with HEV did not cause visible growth impairment or lesions in the chicken embryos. Overall, there was no difference in postinoculation mortality rates among groups sham-inoculated (6/30, 20.0%) or inoculated with live (34/252, 13.4%) or dead (3/ 26, 6.9%) HEV (P = 0.58). The amount of HEV DNA detected in allantoic fluid at 7 dpi in eggs inoculated with live virus was similar to the inoculated dose, indicating that virus propagation in chicken embryos is not efficient. No HEV DNA was detected after 3 dpi in eggs inoculated with dead virus. Inoculation of chicken embryos combined with qualitative PCR can be used for titration of HEV virus stocks and presents a high correlation with in vivo titration using chickens (R² 0.98, P = 0.007). This method may be relevant in countries in which specific-pathogen-free turkeys are unavailable and in which the importation of RP19 cells, the only cell that supports effective propagation of HEV, is not permitted.

Highly pathogenic avian influenza (HPAI) clade 2.3.4.4 viruses from the H5 goose/Guangdong lineage caused a major outbreak in poultry in the United States in 2015. Although the outbreak was controlled, vaccines were considered as an alternative control method, and new vaccines were approved and purchased by the U.S. Department of Agriculture National Veterinary Stockpile for emergency use. In this study, we evaluated the efficacy of two of these vaccines in protecting Pekin ducks (Anas platyrhynchos var. domestica) against challenge with a H5N2 HPAI poultry isolate. A recombinant alphavirus–based vaccine and an inactivated adjuvanted reverse genetics vaccine, both expressing the hemagglutinin gene of a U.S. H5 clade 2.3.4.4 isolate (A/Gyrfalcon/Washington/41088-6/2014 H5N8), were used to immunize the ducks. The vaccines were given either as single vaccination at 2 days of age or in a prime-boost strategy at 2 and 15 days of age. At 32 days of age, all ducks were challenged with A/turkey/Minnesota/12582/15 H5N2 HPAI virus clade 2.3.4.4. All ducks from the nonvaccinated challenge control group became infected and shed virus; one duck in this group presented mild ataxia, and a second duck died. No mortality or clinical signs were observed in vaccinated and challenged ducks, with the exception of one duck presenting with mild ataxia. Both vaccines, regardless of the vaccination strategy used, were immunogenic in ducks and reduced or prevented virus shedding after challenge. In conclusion, good protection against H5Nx infection was achieved in ducks vaccinated with the vaccines examined, which were homologous to the challenge virus, with prime-boost strategies conferring the best protection against infection.

Concurrent use of a colorant during drinking water treatment could contribute to the correct application of fluralaner to poultry. The present studies therefore examined whether the blue-colored drinking water conditioner Vac-Safe has an influence on the stability of fluralaner in water and/or on its efficacy for the control of poultry red mites (PRM).

Laboratory trials showed that fluralaner dissolved at various concentrations in water conditioned with Vac-Safe retained full stability for at least 27 hr at temperatures up to 40 C. Further, a field trial demonstrated that the efficacy of fluralaner in eliminating PRM from two infested houses of laying hens was equal when administered through drinking water with or without Vac-Safe. Consistently in both flocks, after treatment PRM could not be detected and sharp increases in laying percentage and produced egg mass were observed. It was concluded that Vac-Safe does not reduce the stability and treatment efficacy of fluralaner when administered simultaneously through drinking water.

Laying hens (n = 2267) ranging in age from 2 to 4 yr in a study evaluating ovarian cancer prevention were necropsied. Those that died or were culled during the 2-yr study (n = 1591) were necropsied weekly to determine the most probable cause of death or culling and cancer status. Hens surviving until the end of the study (n = 676) were euthanized and necropsied. Hens necropsied before and after a hen with proventricular intussusception served as cohorts (n = 38). Nineteen hens (13 dead, 6 culled) had intussusceptions of the proventriculus into the ventriculus. Mean age of affected hens was 154 wk (range 110–204 wk). None of the hens in the study had an intestinal intussusception, and none of the hens euthanized at the end of the study had a proventricular intussusception. Hens with proventricular intussusceptions were severely emaciated; mean body weights were 1040 and 1736 g for affected and cohort hens, respectively. Necropsy findings included prominent keel, marked muscle atrophy, generalized serous atrophy of fat, no visible proventriculus, esophagus directly entering the ventriculus, and an enlarged, spherical, firm ventriculus, which contained an invaginated, swollen, diffusely ulcerated proventriculus. Eighteen affected hens were anovulatory (94.7%) compared to 27 cohorts (71.1%). Severe, diffuse necrosis and ulceration of the proventricular mucosa was confirmed microscopically, but no etiologic agent was identified. In conclusion, proventricular intussusception of undetermined etiology was identified as a cause of sporadic emaciation, culling, and mortality in older laying hens.