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Avian hosts constitute a natural reservoir for thermophilic Campylobacter species, primarily Campylobacter jejuni and Campylobacter coli, and poultry flocks are frequently colonized in the intestinal tract with high numbers of the organisms. Prevalence rates in poultry, especially in slaughter-age broiler flocks, could reach as high as 100% on some farms. Despite the extensive colonization, Campylobacter is essentially a commensal in birds, although limited evidence has implicated the organism as a poultry pathogen. Although Campylobacter is insignificant for poultry health, it is a leading cause of food-borne gastroenteritis in humans worldwide, and contaminated poultry meat is recognized as the main source for human exposure. Therefore, considerable research efforts have been devoted to the development of interventions to diminish Campylobacter contamination in poultry, with the intention to reduce the burden of food-borne illnesses. During the past decade, significant advance has been made in understanding Campylobacter in poultry. This review summarizes the current knowledge with an emphasis on ecology, antibiotic resistance, and potential pre- and postharvest interventions.

Runting-stunting syndrome (RSS) in poultry has been known for more than 40 years, but the precise etiology remains unknown and a licensed vaccine is consequently not currently available. In order to mitigate the symptoms associated with RSS, a series of experiments was performed to investigate whether a combined bacteriotherapeutic treatment consisting of probiotics, prebiotics, and organic acids could influence the outcome of this disease. Initially two groups of commercial broiler chickens were either left uninoculated or inoculated with filtrate from homogenized intestines of RSS-affected broiler chickens. One group from each of these two challenge groups was treated, with a bacteriotherapeutic regimen. After 12 days chickens were euthanatized, the body weight was measured, and duodenal lesions were enumerated. Five consecutive broiler chicken flocks were then raised either on litter from RSS-affected birds or on fresh wood shavings. Treatment had no beneficial effect on the number and severity of intestinal lesions. There appeared to be a significant build-up of RSS agent(s) in poultry litter, with each consecutive flock placement, independent of bacteriotherapeutic treatment, as more individuals exhibited intestinal lesions on built-up litter in RSS-affected houses (28.9% vs. 44%). While treatment did not appear to consistently reduce intestinal lesions, it did significantly improve the mean body weights (P < 0.05) and uniformity of 12-day-old chickens placed on reused litter in houses in which RSS-infected birds were previously raised. A combination of litter management and bacteriotherapy may be needed to ameliorate the adverse effects of RSS on intestinal health and body weight in broiler chickens.

Several different enteric viruses have been identified as the causes of gastrointestinal infections in poultry. Enteric virus infections are well characterized in poults, but limited studies have been conducted in older birds. The susceptibility of 2-, 7-, 12-, 30-, and 52-wk-old turkeys to turkey coronavirus (TCoV) and turkey astrovirus (TAstV) was evaluated, as well as the effect of combined infection of TAstV and TCoV in 2-wk-old poults and turkey hens. From cloacal swabs and intestines, TCoV was consistently detected by reverse transcriptase-PCR throughout the experimental period (1–21 days postinoculation [DPI]) from all age groups. In contrast, the last detection point of TAstV gradually decreased to 21, 16, and 12 DPI in birds inoculated at 2, 7, and 12 wk of age, respectively, and viral RNA was rarely detected from cloacal swabs or intestinal contents in turkey hens within 3 DPI. Infection with TAstV alone did not affect body weight in poults or egg production in hens. The combined infection of TAstV and TCoV did not induce more severe clinical signs and pathology than the TCoV infection alone. However, a severe prolonged decrease in egg production (about 50%) was observed in turkey hens in the combined infection group compared with a transient egg production drop in the TCoV-infected hens alone. The underlying mechanism regarding the age-related TAstV susceptibility and the pathogenesis of the TAstV and TCoV coinfection in layer hens needs to be further elucidated.

Marek's disease (MD) is a lymphoproliferative disease of domestic chickens that is caused by a highly cell-associated oncogenic α-herpesvirus, Marek's disease virus (MDV). MDV replicates in chicken lymphocytes and establishes a latent infection within CD4 T cells. Clinical signs of MD include depression, crippling, weight loss, bursal/thymic atrophy, neurologic disorders, and rapid onset of T cell lymphomas that infiltrate lymphoid tissues, visceral organs, and peripheral nerves. The cecal tonsils (CTs) are considered the largest lymphoid aggregates of avian gut-associated lymphoid tissue. Along with Peyer's patches, CTs elicit protective immune responses against bacterial and viral pathogens in the intestinal tract of avian species. In this study, we investigated the effect of MDV infection on toll-like receptor (TLR) gene expression in CTs of MD-susceptible (7₂) and resistant (6₃) chicken lines. Real-time PCR gene expression profiling revealed that of the 10 TLRs tested, TLR2A, TLR3, TLR5, and TLR15 displayed significant differential expression patterns at different time points postinoculation. The expression levels of the remaining six genes were minimally affected by MDV infection in either line. Immunohistochemical analysis showed a severe depletion of B cells and CD4 T cells in the CTs of susceptible line at 5 days postinfection (dpi), which recovered by 21 dpi. The destruction of B and T cells in the CTs of the resistant line was minimal at 5 dpi, which also recovered by 21 dpi. A significant infiltration of macrophages was observed after the depletion of B and T cells in the infected birds of both lines that could account for the differential TLR gene expression in the infected birds. The data presented provide further insight into the mechanism of MDV pathogenesis and tissue-specific immunologic responses to viral infection.

Hen diuresis syndrome has emerged over the past 5 yr as a significant cause of mortality in the U.S. broiler breeder industry. The condition affects hens in production and is characterized by transient muscle weakness in the vent region, transient diuresis, and often urate deposits on the skin below the vent. Affected hens are often seen straining to lay an egg, which suggests oviduct contraction is also impaired. Related hen mortality, often reaching 1% or more a week, is believed to be primarily the result of male aggression of the vent region (Turner et al., “Investigating Causes of Excessive Urate Production in Broiler Breeder Hens Associated with Peritonitis and Cannibalism Mortality,” Oral Presentation at The American Association of Avian Pathologists Annual Meeting, p. 139, 2010). The exact association between the cause of mortality and this syndrome is unknown, but it may be the consequence of transient partial to full oviduct prolapse, which predisposes or stimulates cannibalism and aggression. Based on unpublished work done prior to this study (Turner et al., ibid.), the evidence suggests the underlying problem is metabolic. We feel that urine collection and analysis is an essential component to understanding this condition. This study serves as a pilot study for future investigations that attempt to identify the nature and cause of the metabolic disturbance through paired urine and serum collection and analysis. For the purpose of this study, a small sample of 10 affected and 10 unaffected birds was used for sample collection. In order to collect pure urine, the birds were surgically colostomized. Colostomy did prove to be a useful means of collecting urine free of feces, and for the purposes of our study it yielded adequate urine samples for analysis. There were statistically relevant urine values observed. Affected birds had a higher presence of blood in the urine, a lower uric acid excretion rate (mg/hr), higher concentration (mEq/L) of urine Na , and a lower concentration (mEq/L) of urine K than unaffected birds. This pilot study helps to address some of the pitfalls previously associated with colostomy and to determine when collection can begin postoperatively so that we can better understand when and how to begin our sampling in future trials to address the etiology of this condition.

In August 2010, geese in the Meihekou area of Jilin province in China were found to be infected by a pathogen that caused a disease similar to Newcastle disease. To determine the causative agent of the infections, a virus was isolated from liver tissues of infected geese, followed by a pathogenicity determination. The isolated virus was named NDV/White Goose/China/Jilin(Meihekou)/MHK-1/2010. Specific primers were designed to amplify the whole genome of the MHK-1 virus, followed by sequencing and splicing of the entire genome. Sequencing and phylogenetic analysis of MHK-1 showed that the isolate was a virulent strain of Newcastle disease virus. The MHK-1 genome is 15,192 nucleotides long, and it belongs to the class II branch of Newcastle disease viruses, as evidenced by the amino acid sequence (¹¹²R-R-Q-K-R-F¹¹⁷) of the F protein. The hemagglutinin titer was 1:128 to 1:512. The chicken embryo mean death time, the intracerebral pathogenicity index, and the median lethal dose of chicken embryos of MHK-1 were 43 hr, 1.63, and 10⁹/ml, respectively, which revealed that the newly isolated MHK-1 strain is strongly pathogenic to geese.

To evaluate the potential use of an inactivated virus-based vaccine for the control and prevention of the newly emerged duck Tembusu virus infection in China, a duck Tembusu virus isolate, Tembusu-HB, was propagated in 12-day-old duck embryos and inactivated by treatment with formaldehyde. The inactivated viral antigen was emulsified with mineral oil, and five batches of the vaccine were manufactured. The immunogenicity and protection efficacy of the vaccine were evaluated in Beijing ducks and Beijing white geese. Results showed that more than 80% of immunized ducks were protected against virulent virus challenge after two intramuscular or subcutaneous injections of the inactivated vaccine, as evidenced by the negative virus isolation results. The protection is also correlated with a positive virus-specific antibody response as detected by ELISA. In contrast, none of the control ducks and geese had any detectable antibody response. Virus was isolated from all control ducks and geese after virulent virus challenge. Interestingly, a variable level of protection (20%–80%) was observed in Beijing white geese immunized twice with the same batches of vaccine, suggesting a species-specific effect of the vaccine. Overall, the results clearly suggest that the inactivated duck Tembusu virus vaccine is immunogenic and provides protection against virulent virus challenge.

With the aim of assessing the antigenic characteristics of a circulating pool of fowlpox virus (FPV) that exists in the backyard poultry system in India, one of the field isolates generated was characterized by in vitro immunologic techniques. FPV was isolated from clinically positive fowlpox cases (n  =  10) from the Jhargram (West Midnapur district) and Kakdwip (South 24 Pargana district) areas of West Bengal State, India. Initially, FPV-specific PCR was performed for confirmation of the samples. Isolation of FPV was done using embryonated chicken eggs and the choreoallantoic membrane route. Subsequently, FPV antigen was prepared from chicken embryo fibroblast cell culture–adapted field isolate. Biologic transmission of FPV was performed in Rhode Island red chickens experimentally to assess humoral and cell-mediated immune (CMI) responses. High level of anti-FPV antibodies were observed in test birds as assessed by indirect ELISA. Seroreactive polypeptides (B-cell antigens) of FPV antigen with molecular weights of 44.5, 66.5, 75, 90.5, and 99 kDa were detected by western blot analysis. Significant increases in CMI responses were observed in inoculated chickens as assessed by lymphocyte proliferation assay, cytotoxicity assay, and T-cell immunoblotting. The predominant T-cell antigen of FPV detected had a molecular weight of 66.5 kDa. The present study revealed the antigenic characteristics of FPV that exists in backyard poultry system in West Bengal for the first time, thus exploring the rationality of designing future T- and B-cell vaccines against fowlpox.

Laryngotracheitis (LT) is a highly contagious respiratory disease of chickens that produces significant economic losses to the poultry industry. Traditionally, LT has been controlled by administration of modified live vaccines. In recent years, the use of recombinant DNA-derived vaccines using turkey herpesvirus (HVT) and fowlpox virus has expanded, as they protect not only against the vector used but also against LT. However, HVT-based vaccines confer limited protection against challenge, with emergent very virulent plus Marek's disease virus (vv MDV). Serotype 1 vaccines have been proven to be the most efficient against vv MDV. In particular, deletion of oncogene MEQ from the oncogenic vvMDV strain Md5 (BACδMEQ) resulted in a very efficient vaccine against vv MDV. In this work, we have developed two recombinant vaccines against MD and LT by using BACδMEQ as a vector that carries either the LT virus (LTV) gene glycoprotein B (gB; BACΔMEQ-gB) or LTV gene glycoprotein J (gJ; BACδMEQ-gJ). We have evaluated the protection that these recombinant vaccines confer against MD and LT challenge when administered alone or in combination. Our results demonstrated that both bivalent vaccines (BACΔMEQ-gB and BACδMEQ-gJ) replicated in chickens and were safe to use in commercial meat-type chickens bearing maternal antibodies against MDV. BACΔMEQ-gB protected as well as a commercial recombinant (r)HVT-LT vaccine against challenge with LTV. However, BACδMEQ-gJ did not protect adequately against LT challenge or increase protection conferred by BACΔMEQ-gB when administered in combination. On the other hand, both BACΔMEQ-gB and BACδMEQ-gJ, administered alone or in combination, protected better against an early challenge with vv MDV strain 648A than commercial strains of rHVT-LT or CVI988. Our results open a new avenue in the development of recombinant vaccines by using serotype 1 MDV as vectors.

This study related the replication of an H9N2 avian influenza virus in chickens to the induction of host acute immune response after aerosol or intranasal inoculation with the virus. On 1, 2, 4, and 7 days postinoculation (dpi), oropharyngeal swabs and tissue specimens of trachea, lungs, spleen, and cecal tonsils were collected for quantification of viral RNA. Expression of cytokine genes in lungs, spleen, and cecal tonsils was quantified by reverse transcriptase-PCR. Virus was detected in all oropharyngeal swabs up to 4 dpi in chickens from both inoculation groups. However, virus was detected more frequently (P < 0.05) and in higher titers (1–4 log difference) in specimens of trachea and lungs from the group exposed to aerosols than from the group given intranasal drops. In accord with viral replication findings, expressions of cytokine genes interleukin (IL)-1β (on 2 and 7 dpi), IL-6 (on 2 dpi), and interferon (IFN)-γ (on 2 and 4 dpi) were up-regulated to a significantly higher level (P < 0.05) in lung tissue specimens from the group exposed to virus aerosol than from controls that were given saline intranasally. Only IFN-γ on 1 dpi was up-regulated (P < 0.05) above that of controls in lung tissue specimens from the group given intranasal drops of virus. In comparison, replication of the virus and induction of IL-1β and IL-6 genes were limited in spleen and cecal tonsil tissue specimens from both groups of chickens inoculated with the virus. These findings indicate that virus administered in aerosols was more efficient than virus administered as intranasal drops, in infecting the lower respiratory tract and in inducing the activity of the cytokine genes. The intense respiratory infection caused by virus aerosols might increase the shedding and transmission of the H9N2 virus in chickens.

A major limiting issue of bacterial ghost technology involves the stable maintenance of Phix174 lysis gene E expression. Unwanted leaky expression of gene E in the absence of induction temperature results in reduced biomass production of host bacterium, consequently leading to the lower yield of bacterial ghost. To mitigate the leaky expression status of lysis gene E, we utilized a novel E-lysis system in which gene E is located between sense λpR promoter with a CI857 regulator and antisense P_araBAD promoter with the AraC regulator. In the presence of l-arabinose at 28 C, unwanted transcription of lysis gene E from λpR promoter is repressed by a simultaneous transcription event from P_araBAD promoter by means of anti-sense RNA-mediated inhibition. Tight repression of lysis gene E in the absence of induction temperature resulted in higher bacterial cell number in culture suspension and, consequently, higher production of Salmonella Gallinarum (SG) ghost biomass. The safety and protective efficacy of the SG ghost vaccine were further examined in chickens. All of the immunized chickens showed significantly higher mucosal and systemic antibody responses accompanied by a potent antigen-specific lymphocyte proliferative response. Vaccination of chickens with SG ghost preparation offered efficient protection against wild-type SG challenge.

Parasitic infections caused by Eimeria species are responsible for most economic losses in poultry production. Prevalence studies can adequately assist the design of prophylaxis strategies for disease control. Therefore, stool samples from 251 flocks of broilers from 28 to 48 days old were collected in 21 municipalities in the state of Santa Catarina, Brazil, to detect and examine the prevalence of Eimeria acervulina, Eimeria maxima, Eimeria tenella, Eimeria mitis, Eimeria praecox, Eimeria necatrix, and Eimeria brunetti. The oocysts were recovered and quantified, and the species were identified by a multiplex PCR technique. Amplicons of seven Eimeria species originating from the PCR-positive samples were cloned. Microscopy studies demonstrated that 96% of the farms were positive for the Eimeria. Seven species were identified, as follows: E. maxima (63.7%) and E. acervulina (63.3%) were the most prevalent species, followed by E. tenella (54.6%), E. mitis (38.6%), E. praecox (25.1%), E. necatrix (24.3%), and E. brunetti (13.1%). The average number of species detected per farm was 2.96, and the most common were E. acervulina, E. maxima, and E. tenella (9.16%). The sequencing of the clones confirmed the specificity and effectiveness of multiplex PCR for the identification of seven species of Eimeria, so this tool can be useful in studying circulating species in poultry farms, thereby assisting prophylactic measures against coccidiosis.

Muscovy duck reovirus (MDRV) causes high morbidity and mortality in ducklings. However, the molecular basis for pathogenesis of this virus remains poorly understood, and the complete genome sequence of Muscovy duck is lacking. Here we report the transcriptome profile of Muscovy ducks in response to MDRV infection. RNA sequencing technology was employed to obtain a representative complement of transcripts from the spleen of ducklings, and then differential gene expression was analyzed between MDRV-YB strain infected ducks and noninfected ducks. This analysis generated 65,536 unigenes. Of these, 6458 genes were found to be significantly differentially expressed between the infected and noninfected groups. The symptom and pathology of ducks infected with MDRV-YB was correlated with the greater proportion of decreased expression genes (4906) than increased expression (1552) level. Gene ontology analysis assigned differentially expressed genes to the categories: “biological processes,” “cellular functions,” and “molecular functions.” Differentially expressed genes involved in the innate immune system were analyzed further, and 128 of these genes showed decreased expression and 86 showed increased expression between the infected and noninfected groups. These genes represented the Janus kinase–signal transducer and activator of transcription signaling pathway, and the retinoic acid-inducible gene I (RIG-I)–like and Toll-like receptor (TLR) signaling pathways and included interferon (IFN) α, IFNγ, interleukin 6, RIG-I, and TLR4. The data were verified by SYBR fluorescence quantitative polymerase chain reaction (SYBR-qPCR). Our findings offer new insight into the host immune response to MDRV infection.

Salmonella contamination of laying hen flocks and shell eggs is associated with various management and environmental factors. Foodborne outbreaks of human salmonellosis have been traced back to consumption of Salmonella-contaminated shell eggs. In the present study, a systematic literature review was conducted to identify and provide an evidence-based overview of potential risk factors of Salmonella contamination of laying hens, layer premises, and shell eggs. This systematic literature search was conducted using AGRICOLA, CAB Abstracts, and PubMed databases. Observational studies that identified risk factors for Salmonella contamination of layer flocks and shell eggs were selected, and best evidence was synthesized to summarize the results. Altogether, 13 cross-sectional studies and four longitudinal studies published in English were included in the review. Evidence scores were assigned based on the study design and quality of the study to grade the evidence level. The strength of association of a risk factor was determined according to the odds ratios. In this systematic review, the presence of previous Salmonella infection, absence of cleaning and disinfection, presence of rodents, induced molting, larger flock size (>30,000 hens), multiage management, cage housing systems, in-line egg processing, rearing pullets on the floor, pests with access to feed prior to movement to the feed trough, visitors allowed in the layer houses, and trucks near farms and air inlets were identified as the risk factors associated with Salmonella contamination of laying hen premises, whereas high level of manure contamination, middle and late phase of production, high degree of egg-handling equipment contamination, flock size of >30,000, and egg production rate of >96% were identified as the risk factors associated with Salmonella contamination of shell eggs. These risk factors demonstrated strong to moderate evidence of association with Salmonella contamination of laying hens and shell eggs. Eggshells testing positive for Salmonella were 59 times higher when fecal samples were positive and nine times higher when floor dust samples were positive. Risk factors associated with Salmonella Enteritidis infection in laying hens were flock size, housing system, and farms with hens of different ages. As a summary, this systematic review demonstrated that Salmonella contamination of laying hen flocks and shell eggs in layer production systems is multifactorial. This study provides a knowledge base for the implementation of targeted intervention strategies to control Salmonella contamination of laying hen flocks and shell eggs.

Missouri, United States, is located within the Mississippi Migratory Bird Flyway where wild waterfowl stop to feed and rest during migration and, weather permitting, to overwinter. Historically, Missouri has experienced sporadic influenza A virus (IAV) outbreaks in poultry and commercial swine. The introduction of IAVs from wild, migratory waterfowl is one possible source for the IAV, IAV genomic segments, or both involved in these outbreaks in key agricultural species. During 2005 through 2013, 3984 cloacal swabs were collected from hunter-harvested waterfowl in Missouri as part of an active IAV surveillance effort. Twenty-four avian species were represented in the sample population and 108 (2.7%) of the samples tested positive for IAV recovery. These IAV isolates represented 12 HA and nine NA subtypes and at least 27 distinct HA–NA combinations. An H14 IAV isolate recovered in Missouri during the sample period provided evidence for further establishment of the H14 subtype in North American wild waterfowl and gave proof that the previously rare subtype is more genetically diverse than previously detected. The present surveillance effort also produced IAV isolates that were genomically linked to the highly pathogenic H7N3 IAV strain that emerged in 2012 and caused severe disease in Mexico's domestic poultry. The presence of antigenically diverse IAV's circulating in wild waterfowl in the vicinity of commercial poultry and swine, along with the association of several wild-bird–lineage IAV genomic segments in viruses infecting poultry in North America, justifies continued attention to biosecurity efforts in food animal production systems and ongoing active IAV surveillance in wild birds.

This study was performed to investigate the prevalence and to characterize the genetic diversity of Histomonas meleagridis isolates in chickens in southern Vietnam. A total of 194 chickens, randomly selected from 18 backyard and 18 commercial flocks, were screened for H. meleagridis infection using both macroscopic diagnosis and an 18S rRNA gene-based PCR method. Overall, 12.9% of birds, representing 19 flocks, showed gross lesions typical for histomonosis whereas 25.3% of the birds from 29 flocks were positive by PCR assay. Following initial diagnostic approaches, H. meleagridis-positive samples were further analyzed by sequencing three different genomic loci; the 18S rRNA, alpha-actinin1, and rpb1. Thirteen samples from 12 flocks were genetically identified as H. meleagridis, demonstrating a flock and sample prevalence of 33.3% and 6.7%, respectively. There was no significant difference in prevalence between different farm types, age groups, and seasonality. Genetic analysis demonstrated minor heterogeneity of Vietnamese isolates with 99% homology to H. meleagridis sequences from the database. This is the first survey of the prevalence and genetic characterization of H. meleagridis in chickens in Vietnam.

An unusual outbreak of chlamydiosis was diagnosed in 15,000, 13-wk-old organically grown turkeys housed in a semiconfinement housing system. The disease was characterized by unilateral or bilateral swelling above the eye due to mild-to-severe inflammation of the nasal glands in 3%–5% of the birds. Except for a slight drop in feed and water consumption, the birds did not exhibit any respiratory signs, morbidity, and mortality. Chlamydiosis in the turkeys was confirmed by immunofluorescence, immunohistochemistry, and PCR assay of the nasal glands. Other samples such as conjunctiva, lungs, air sacs, heart, liver, spleen, and feces were negative for chlamydia by florescence antibody test in birds submitted over several weeks. Chlamydia psittaci strain B was isolated in chicken egg embryos and typed by multilocus sequence variable number of tandem repeats analysis, multilocus sequence typing, and ompA gene sequencing as a CP3-like strain. This is the first report of a naturally occurring chlamydiosis affecting the nasal glands in turkeys.

Genetic selection for fast growth can affect the ability of male turkeys to cope with stressors, resulting in decreased immunity to opportunistic bacterial infection. The purpose of the current study was to compare the effects of ascorbic acid (AA) on the stress response and resistance to Escherichia coli challenge of birds selected for increased 16-wk body weight (BW; F-line) with their random-bred parent line (RBC2). Male turkeys were raised in duplicate floor pens in a two line × two AA treatment × two stress challenge (SC) design. At 5 wk of age, AA (1200 ppm) was provided in drinking water for a 24-hr period, during which all birds were weighed. After AA treatment, the SC group was subjected to a transport stress protocol. Six hours after the start of transport, SC birds were also inoculated in the thoracic air sac with 1 × 10⁴ colony-forming units of E. coli. The following morning four birds from each pen were bled, and all birds were weighed and necropsied 2 days later. BW and gain after SC were decreased in the F-line but not the RBC2 line, and there were no AA effects on BW. The weight of the bursa of Fabricius relative to BW was higher in the RBC2 line than in the F-line, was decreased by SC, and was not affected by AA. The heterophil∶lymphocyte ratio was higher in the SC F-line as compared to the SC RBC2 and was decreased by AA only in the SC F-line. Corticosterone (C) levels were increased by SC only in the F-line, and AA decreased C levels only in the RBC2 line. Airsacculitis scores were increased in the F-line SC birds. The challenge strain of E. coli was only detected in the air sac and liver of the AA-treated F-line SC birds and in the liver of the no-AA F-line birds. These results suggest that SC at 5 wk of age had a more deleterious effect on the fast-growing F-line than on its parent line and that AA may have increased susceptibility to colibacillosis in the SC F-line birds.

A low pathogenic avian influenza virus was identified from free-living birds (mynah, Acridotheres tristis) of the starling family. Virus was isolated by inoculation of homogenized suspension from lung, tracheal, spleen, and cloacal swabs into the allantoic cavity of embryonated chicken eggs. Subtype of the isolate was characterized as H9N2 by hemagglutination inhibition test using monospecific chicken antisera to a wide range of influenza reference strain. Pathogenicity of the isolate was determined by intravenous pathogenicity index. The virus was reisolated from experimentally infected chicken. Additionally, the isolate was subjected to reverse transcriptase PCR using partial hemagglutinin (HA) gene-specific primers and yielded an amplicon of 487 bp. HA gene sequence analysis revealed 99% sequence homology among mynah and chicken isolates from Oman. On phylogenetic analysis, isolates from mynah (A/mynnah/Oman/AIVS6/2005) and chicken (A/chicken/Oman/AIVS3/2006; A/chicken/Oman/AIVS7/2006) clustered together tightly, indicating these free-flying birds may be a source of introduction of H9N2 subtype in poultry bird in Oman. Moreover, the HA gene of H9N2 isolates from Oman resembled those of viruses of the G1-like lineage and were very similar to those from United Arab Emirates.

This case report describes a recent botulism outbreak in commercial laying hens with a history of increased mortality and flaccid paralysis. Routine diagnostic gross examination and microscopy from seven hens were inconclusive, but botulinum neurotoxin (BoNT) in peripheral blood was neutralized with both type C and type D antitoxins in the mouse bioassay. During a farm visit, 10 additional hens from a 34-wk-old flock on the farm were selected for clinical examination and further sampling. Nine hens were observed in sternal recumbency, with flaccid paralysis of the neck, drooping wings and tail, inability to escape, and bilateral ptosis, and one hen showed nonspecific clinical signs. Samples from cecum and liver were collected, and the gene coding for BoNT was detected by PCR in all 10 cecal samples and in four of the liver samples. Clostridium botulinum mosaic type C/D was isolated from 5 out of 10 hens from either cecum or liver, and the isolates were subjected to pulsed-field gel electrophoresis subtyping. All five isolates produced the same banding pattern, which was identical or showed >90% similarity to isolates from three different outbreaks on broiler farms in Sweden and Denmark during the 2007–10 period. However, they were clearly distinguishable from the predominantly reported pulsotype associated with avian botulism outbreaks in Europe. The authors are unaware of any previous report of C. botulinum mosaic type C/D isolates from laying hens.

The β-nicotinamide adenine dinucleotide (NAD) requirement has been considered to be essential for the isolation of the causal agent of infectious coryza, Avibacterium paragallinarum. Nevertheless, NAD-independent reports from South Africa and Mexico dismissed this paradigm. It is now accepted that both NAD-dependent and NAD-independent agents are able to cause infectious coryza and thus belong to the species A. paragallinarum. Here, we report for the first time in Peru a NAD-independent isolate from broiler chickens with typical signs of infectious coryza that have received a trivalent inactivated vaccine against infectious coryza. The isolate was identified based on its morphology, biochemical and serologic tests, and PCR results. Partial 16S rRNA gene sequence analysis confirmed the isolate as A. paragallinarum. There have been no cases of NAD-independent A. paragallinarum isolates reported in South America. Increasing reports around the world highlight not only the need to reconsider the in vitro nutritional requirements of this species for its correct isolation but also the cross-protection conferred by commercial infectious coryza vaccines against NAD-independent isolates.

In April 2014 an outbreak of low pathogenic avian influenza H5N8 North American genetic lineage was diagnosed in a commercial quail operation in Stanislaus County, California. Sudden increase in mortality prompted the submission of 20 Japanese quail hens (Coturnix c. japonica) to the California Animal Health and Food Safety Laboratory, Turlock Branch. Oropharyngeal and cloacal swabs tested positive for influenza A virus H5N8 by real-time reverse transcription–polymerase chain reaction. The virus was subsequently isolated. In vivo assay and sequencing of the hemagglutinin protein cleavage site classified the virus as a North American genetic lineage of low pathogenicity for chickens. Following the diagnosis, a rapid and coordinated response took place to contain the outbreak. The affected premise was depopulated, cleaned, and disinfected. Three areas from the affected premises—a 3 kilometer (km) radius (High Risk Zone), a 3–10 km area (Buffer Zone), and a 10–20 km (Surveillance Zone)—were established for avian influenza testing of commercial and noncommercial poultry operations. Surveillance testing and rapid control measures were successful in the control and eradication of the outbreak and revealed no area of spread of the virus from the index flock. This report describes the history, diagnosis, surveillance, and control measures applied to manage this outbreak.