Table Egg Layer Research Articles

Comparison of bone quality in table egg layers by quantitative ultrasound, 3-point-bending test, and ash content

Comparison of bone quality in table egg layers by quantitative ultrasound, 3-point-bending test, and ash content

Egg Drop Syndrome 76 in a U.S. Broiler Breeder Flock.

A four-house broiler breeder farm of approximately 35-wk-old hens was diagnosed with egg drop syndrome (EDS'76) utilizing PCR and hemagglutination inhibition (HI) testing. Based on communication with local practitioners, the geographic area near where this flock was located had numerous EDS'76 cases in table egg layers at the time of diagnosis. An egg production drop was seen in the broiler breeder flock over a 7-day period, which prompted an investigation. During this investigation, a significant number of shell-less, wrinkled, and pale eggs were noted, but no increases in mortality or respiratory signs were observed. The disease and subsequent production drops spread horizontally across the farm over a 5-wk period. Production returned to approximately the breed standard egg production 4 wk after initial egg production drop. However, hatching egg utilization continued to be reduced for another 2 wk because of the number of thin-shelled eggs. A similar pattern of drop in egg production and subsequent return was observed in the other houses. No significant lesions were noted in the tissues submitted for histopathology. Differential diagnoses that could cause shell abnormalities and egg production drops were ruled out by submitting appropriate samples for diagnostic investigation. Egg drop syndrome 76 PCR was performed on shell-less eggs and EDS'76 was detected by PCR at two separate laboratories. Subsequently, serum was submitted for HI and positive results were found in each house as they showed egg production drop concurrent with shell abnormalities. At the time the flocks returned to breed standard production, EDS'76 titers were consistent with a uniformly exposed and seroconverted flock. The authors suspect that immunosuppression as pullets played a role in this flock being impacted by EDS. No other farms in the company's system were observed with production drops or shell abnormalities similar to this case.

Laser Capture Microdissection, Culture Analysis, and Bacterial Sequencing to Evaluate the Microbiota of Focal Duodenal Necrosis in Egg Layers.

Focal duodenal necrosis (FDN) is a common intestinal disease of table egg layers. In this research we aimed to identify the bacteria commonly found in FDN lesions as seen with histopathological analysis. Fifty-nine ethanol-fixed duodenum samples were collected from egg layers on eight FDN-affected farms, and 42 samples had typical FDN lesions. Excision of bacteria-containing lesions using laser capture microdissection was performed, followed by 16S rRNA gene sequencing of extracted DNA for bacterial identification. Bacterial sequencing analysis revealed no consistent bacterial species identified from samples with FDN. However, analysis of the relative phylum abundance revealed differences in the duodenal microbiota between layers with FDN and healthy birds. There were differences in the abundance of Proteobacteria, Firmicutes, and Actinobacteria between FDN-positive and FDN-negative control samples compatible with intestinal dysbiosis. In addition, 10 duodenal samples with FDN lesions were collected for bacteriological analysis, yielding 47 colonies on tryptone soy agar, MacConkey agar, and blood agar plates. Using 16S rRNA gene PCR, 39/47 (53.8%) colonies were identified as Escherichia coli. PCR for E. coli virulence genes identified 21/39 (53.8%) E. coli isolates as avian pathogenic E. coli-like. PCR analysis for 19 E. coli virulence genes associated with intestinal disease strains including inflammatory bowel disease found 11/39 (28.2%) isolates containing more than 10 of these virulence genes. In conclusion, FDN appears to be a multifactorial inflammatory intestinal disease associated with intestinal dysbiosis, and Gram-negative bacteria including E. coli may contribute to the pathogenesis of this disease.

Hemorrhagic Hepatopathy in Broiler Breeder Pullets: Gross and Microscopic Pathology and Factors Associated with Incidence.

Hemorrhagic hepatopathy is a syndrome reported in layer pullets resulting in mortality and lesions including hepatic, splenic, and intestinal necrosis; hepatic and splenic enlargement; hemorrhages; amyloidosis of the muscle, spleen, and liver; accumulation of noncoagulated hemorrhagic fluid in the coelom; and frequently, granulomatous myositis at bacterin injection sites. The syndrome is characterized in the literature in table egg layer pullets and is thought to be associated with the administration of bacterin vaccines, namely, frequently Salmonella enterica subsp. enterica bacterins. Hemorrhagic hepatopathy is recognized by industry veterinarians as also occurring infrequently in broiler breeder pullets in the United States. As the condition is likely due to an inflammatory process in response to bacterial lipopolysaccharide inoculation, it is important to characterize both the pathologic changes and predisposing factors for the condition in broiler breeds, which are immunologically different from table egg layer breeds. In this study, we characterize the gross and microscopic lesions observed in a series of diagnostic laboratory cases of hemorrhagic hepatopathy in broiler breeder pullets and suggest a possible pathophysiology for the condition. Additionally, we report results from a case survey of the United States broiler industry that suggest that the condition is due to a reaction to bacterin vaccination and that certain bacterin products may predispose pullet flocks to develop the condition. Although further research is indicated, these findings establish hemorrhagic hepatopathy as a pathologic condition of broiler breeder pullets and may aid in the diagnosis and prevention of the syndrome.

Occurrence and Characterization of Salmonella Isolated from Table Egg Layer Farming Environments in Western Australia and Insights into Biosecurity and Egg Handling Practices.

The aim of this study was to investigate the occurrence and distribution of Salmonella in commercial layer farming environments of 26 flocks belonging to seven egg businesses (free-range and barn-laid) in Western Australia (WA). Between November 2017 and June 2018, a total of 265 environmental samples of dust, feed, water, pooled feces, and boot swabs were tested for detection of Salmonella according to standard culture-based methods. Isolates were assayed for serovar and subtyped by multilocus sequence typing (MLST). Salmonella spp. were recovered from 35% (93/265) of all tested samples. Dust (53.8%, 28/52) and pooled fecal (54.5%, 18/33) samples provided the highest Salmonella recovery rates. Nine different Salmonella serovars were characterized across the positive (n = 93) environmental samples, of which S. Typhimurium (60/93, 64.5%) and S. Infantis (21/93, 22.5%) were the most prevalent. MLST revealed that all S. Typhimurium isolates were of sequence type ST-19. Microbiological screening of Salmonella was not routinely practiced in any of the surveyed egg businesses. Some of the egg businesses exhibited variable levels of compliance with basic biosecurity measures as well as high-risk egg handling practices. Egg businesses in WA should be encouraged to adopt a voluntary program of environmental sampling and verification testing for Salmonella. Such voluntary programs will aid in supporting solutions for the management of this pathogen in the human food chain.

Profiling of Egg Layer Flocks Affected with Focal Duodenal Necrosis Based on a Questionnaire Survey.

Focal duodenal necrosis (FDN) is an intestinal disease of egg-laying chickens, characterized by multifocal mucosal erosions in the duodenal loop and proximal jejunum. It is currently considered by the Association of Veterinarians in Egg Production and the United States Animal Health Association as one of the top five disease concerns of the table egg industry in the United States. Previous studies have associated this condition with Clostridium species. The purpose of this study was to investigate the epidemiologic characteristics of table egg layer flocks affected with FDN. An online questionnaire was distributed to commercial layer operations in different states in the United States. Layer farms that had diagnosed FDN within the past 12 mo were surveyed. The questionnaire had 45 questions about management, nutrition, housing, and methods for disease prevention and control. Thirty-seven surveys were sent and 21 were completed, which represents a response rate of 56.7%. The survey results showed the presence of FDN in five egg-layer genetic lines or breed crosses of different ages, with most cases reported between 30-39 wk of age. The pullets were cage-reared in all affected flocks and the majority of flocks in production were housed in traditional cages. Most of the FDN-affected flocks received more than 12 different feed formulations from pre-lay to 60 wk of age. Distiller's dried grain with solubles was a common ingredient added to the feed in the majority of affected flocks, and all flocks were provided with limestone as a calcium source for egg production. Most surveys reported that coccidiosis and roundworm parasitism were not problems in affected flocks in production; however, pests such as flies and rodents were reported as problems in most affected flocks. Additionally, most affected farms never washed feeders, cages, and houses before disinfection, which may not be sufficient to prevent the persistency and transmission of the causative agent of FDN. In conclusion, several management practices that have been associated with enteric disease, including clostridial-associated enteritis, were described by the majority of FDN-affected flocks. Additional studies are needed to determine if management and health practices identified in this survey represent risk factors for FDN.

Characterization of prevalent bacterial pathogens associated with pododermatitis in table egg layers

ABSTRACT Pododermatitis has been observed in several layer flocks in Denmark during 2015. The aetiology is complex, including litter quality, nutrition and management. Bacterial pathogens associated with pododermatitis, however, have not received much attention. The aim of the present study was, therefore, to identify 106 bacterial isolates obtained from pododermatitis in table egg layers in addition to five isolates from spleen/bursa presternalis. Isolates were obtained from layers from six affected flocks. All isolates were identified by standard bacterial methods, species-specific PCRs, 16S rRNA sequencing or matrix-assisted laser desorption-ionization identification. Staphylococcus aureus and Enterococcus faecalis made up 75/111 (68%) and 15/111 (14%) of the isolates from pododermatitis, respectively; the remaining isolates represented Escherichia coli (10), Staphylococcus hyicus (5), Gallibacterium anatis (3), Trueperella pyogenes (2) and Aerococcus urinaeequi (1). All isolates of S. aureus were spa-typed. Spa-type t8646 and t002 made up 72% and 26% of the S. aureus isolates, respectively. The same types were also demonstrated from spleen/bursa presternalis. The same or closely related spa-types were found among 6/11 sepsis-affected day-old chicks included for comparison, indicating that these types of S. aureus are ubiquitous pathogens in poultry. In contrast, isolates of E. faecalis and E. coli showed major population diversity. In conclusion, the results suggest that S. aureus is a major pathogen associated with pododermatitis abscesses, which could be from a common source, whereas the diversity among the E. faecalis and E. coli populations suggests that these bacteria might originate from multiple sources.

Revenue Impacts of the 2015 Avian Influenza Virus Outbreak on the United States Table Egg Wholesalers

From December 2014 to June 2015, the U.S. poultry industry experienced an outbreak of highly pathogenic avian influenza (AI), resulting in considerable bird depopulations. Both turkey and egg producers were impacted and farms affected faced losses from costs of bird disposal and farm repopulation. This study isolates the table egg subsector of the poultry industry and looks at the revenue impact of the AI outbreak at the wholesale level. To determine this revenue impact, a vector error correction model (VECM) was defined and used to generate the counterfactual revenue during the time period the outbreak occurred. This counterfactual revenue was compared to the actual revenue observed during that time period and the difference is the revenue impact due to the outbreak, ceteris paribus. Additionally, machine learning algorithms, using innovations from the VECM, allowed determining causal relationships in contemporaneous time among the variables considered within the industry. The model was developed using eight variables, the number of table egg layers on farm during a month and eggs, egg price, feed input prices, retail pork and beef prices, and real disposable personal income. When the counterfactual revenue was compared to the actual revenue from December 2014 to June 2015, a gain of about $676 million to wholesalers was determined to be attributed to the outbreak. Additionally, analysis of contemporaneous relationships of innovations, as shown by directed acyclic graphs, indicated that innovations in egg price are independent of innovations in direct production quantities, number of layers on farm and eggs, but are affected by production costs such as feed input costs. These results can be accounted for by various factors including the inelasticity of egg price and the imperfectly competitive behavior of the wholesalers and time delays in layer production. Future studies can use price transmission principles to expand this study and identify AI outbreak impacts at the consumer and producer levels.

Prevalence of Salmonella spp. in environmental samples from table egg barns in Alberta

ABSTRACTSome Salmonella spp. are zoonotic, a frequent cause of foodborne illness in Canada, and known to infect humans through contaminated poultry and poultry products. Certain serotypes of Salmonella spp. have been demonstrated to be vertically transmitted from hen to egg. The incidence of Salmonella spp. isolation in the flock has been correlated to its isolation from the environment. Twenty-one producers were enrolled in this study to examine the occurrence of Salmonella spp. in 48 table egg layer flocks housed in 35 barns in Alberta. The purpose of this study was to: (i) identify Salmonella serotypes isolated from the environment of table egg layer facilities in Alberta and (ii) record the prevalence of Salmonella spp. across eight defined environmental sampling points. Salmonella spp. were isolated from the environment of 20/35 barns representing 29/48 flocks. The most common serotypes isolated were S. Heidelberg, S. Kentucky and S. Mbandaka. The order of most to least contaminated sample location was manure belts (54.1%), feeders (47.9%), feed motors (45.8%), egg belts and walls (41.7%), fans (35.0%), cage bottoms (31.3%) and lobbies (27.1%). Salmonella spp. were isolated from 7/7 barns post cleaning and disinfection, demonstrating the persistence of this organism in the environment and the need for effective eradication protocols.

H5N2 Highly Pathogenic Avian Influenza Viruses from the US 2014-2015 outbreak have an unusually long pre-clinical period in turkeys.

BackgroundFrom December 2014 through June 2015, the US experienced the most costly highly pathogenic avian influenza (HPAI) outbreak to date. Most cases in commercial poultry were caused by an H5N2 strain which was a reassortant with 5 Eurasian lineage genes, including a clade 2.3.4.4 goose/Guangdong/1996 lineage hemagglutinin, and 3 genes from North American wild waterfowl low pathogenicity avian influenza viruses. The outbreak primarily affected turkeys and table-egg layer type chickens. Three isolates were selected for characterization in turkeys: the US index isolate from December 2014 (A/northern pintail/WA/40964/2014), and two poultry isolates from April 2015 (A/chicken/IA/13388/2015 and A/turkey/MN/12528/2015).ResultsFour week old broad-breasted white turkeys were inoculated with one of three doses (102, 104 or 106 50% egg infectious doses [EID50] per bird) of each of the isolates to evaluate infectious dose and pathogenesis. The mean bird infectious dose of A/northern pintail/WA/40964/2014 and A/turkey/MN/12528/2015 was 105 EID50 per bird, but was 103 EID50 per bird for A/chicken/IA/13388/2015, suggesting the latter had greater adaptation to gallinaceous birds. All three isolates had unusually long mean death time of 5.3–5.9 days post challenge, and the primary clinical signs were severe lethargy and neurological signs which started no more than 24 h before death (the average pre-clinical period was 4 days). Infected turkeys also shed high levels of virus by both the oropharyngeal and cloacal routes.ConclusionsThe unusually long mean death times, high levels of virus in feces, and increased adaptation of the later viruses may have contributed to the rapid spread of the virus during the peak of the outbreak.

Factors Associated with Highly Pathogenic Avian Influenza H5N2 Infection on Table-Egg Layer Farms in the Midwestern United States, 2015.

A case-control study was conducted among commercial table-egg layer and pullet operations in Iowa and Nebraska, United States, to investigate potential risk factors for infection with highly pathogenic avian influenza (HPAI) H5N2. A questionnaire was developed and administered to 28 case farms and 31 control farms. Data were collected at the farm and barn levels, enabling two separate analyses to be performed-the first a farm-level comparison of case farms vs. control farms, and the second a barn-level comparison between case barns on case farms and control barns on control farms. Multivariable logistic regression models were fit using a forward-selection procedure. Key risk factors identified were farm location in an existing control zone, rendering and garbage trucks coming near barns, dead-bird disposal located near barns, and visits by a company service person. Variables associated with a decreased risk of infection included visitors changing clothing, cleaning and disinfecting a hard-surface barn entryway, and ceiling/eaves ventilation in barns.

Association of Beta2-Positive Clostridium perfringens Type A With Focal Duodenal Necrosis in Egg-Laying Chickens in the United States.

Focal duodenal necrosis (FDN) is a poorly understood intestinal disease of egg layers, and has been associated with drops in egg production and decreased egg weights. The etiology of this disease is still unknown, but the condition has been associated with Clostridium colinum and Clostridium perfringens. In order to investigate the etiology, duodenal samples were taken from hens with FDN. The hens originated from table egg layer farms in three states. The samples were examined by histopathology, bacteriology, and immunohistochemistry. Macroscopically, all samples contained focal to multifocal, variably sized, reddened or brownish gray areas of mucosal erosion. Histopathology revealed mild to severe heterophilic and lymphoplasmacytic enteritis with loss of enterocytes at the villous tips, luminal fibrinonecrotic exudate, and variable numbers of Gram-positive and Gram-negative rod-shaped bacteria within the lesions in 16/30 samples. Clostridium perfringens was isolated by anaerobic bacteriology from 4/13 samples that had characteristic microscopic lesions of FDN. Polymerase chain reaction (PCR) revealed that all four isolates were Type A C. perfringens, positive for beta2 gene and negative for necrotic enteritis toxin B and enterotoxin genes. PCR for Clostridium colinum applied to DNA extracted from frozen intestinal samples yielded negative results in 14/14 duodenal samples. Immunohistochemistry (IHC) for 7C. perfringens, alpha and beta2 toxins stained a few to numerous long rod-shaped bacteria present in the lesions. IHC for alpha and beta2 toxins also stained enterocytes at the villous tips, inflammatory cells in the lamina propria, as well as degenerated and sloughed enterocytes present within the luminal exudate. These findings suggest that C. perfringens may play a role in the development of FDN. Experimental challenge studies with these isolates still need to be performed in order to reproduce the disease and fulfill Koch's postulates.

Validation of Single and Pooled Manure Drag Swabs for the Detection of Salmonella Serovar Enteritidis in Commercial Poultry Houses.

Single swabs (cultured individually) are currently used in the Food and Drug Administration (FDA) official method for sampling the environment of commercial laying hens for the detection of Salmonella enterica ssp. serovar Enteritidis (Salmonella Enteritidis). The FDA has also granted provisional acceptance of the National Poultry Improvement Plan's (NPIP) Salmonella isolation and identification methodology for samples taken from table-egg layer flock environments. The NPIP method, as with the FDA method, requires single-swab culturing for the environmental sampling of laying houses for Salmonella Enteritidis. The FDA culture protocol requires a multistep culture enrichment broth, and it is more labor intensive than the NPIP culture protocol, which requires a single enrichment broth. The main objective of this study was to compare the FDA single-swab culturing protocol with that of the NPIP culturing protocol but using a four-swab pool scheme. Single and multi-laboratory testing of replicate manure drag swab sets (n = 525 and 672, respectively) collected from a Salmonella Enteritidis-free commercial poultry flock was performed by artificially contaminating swabs with either Salmonella Enteritidis phage type 4, 8, or 13a at one of two inoculation levels: low, x¯ = 2.5 CFU (range 2.5-2.7), or medium, x¯ = 10.0 CFU (range 7.5-12). For each replicate, a single swab (inoculated), sets of two swabs (one inoculated and one uninoculated), and sets of four swabs (one inoculated and three uninoculated), testing was conducted using the FDA or NPIP culture method. For swabs inoculated with phage type 8, the NPIP method was more efficient (P < 0.05) for all swab sets at both inoculation levels than the reference method. The single swabs in the NPIP method were significantly (P < 0.05) better than four-pool swabs in detecting Salmonella Enteritidis at the lower inoculation level. In the collaborative study (n = 13 labs) using Salmonella Enteritidis phage type 13a inoculated swabs, there was no significant difference (P > 0.05) between the FDA method (single swabs) and the pooled NPIP method (four-pool swabs). The study concludes that the pooled NPIP method is not significantly different from the FDA method for the detection of Salmonella Enteritidis in drag swabs in commercial poultry laying houses. Consequently based on the FDA's Salmonella Enteritidis rule for equivalency of different methods, the pooled NPIP method should be considered equivalent. Furthermore, the pooled NPIP method was more efficient and cost effective.

Impact of route of exposure and challenge dose on the pathogenesis of H7N9 low pathogenicity avian influenza virus in chickens

H7N9 influenza A first caused human infections in early 2013 in China. Virus genetics, histories of patient exposures to poultry, and previous experimental studies suggest the source of the virus is a domestic avian species, such as chickens. In order to better understand the ecology of this H7N9 in chickens, we evaluated the infectious dose and pathogenesis of A/Anhui/1/2013 H7N9 in two common breeds of chickens, White Leghorns (table-egg layers) and White Plymouth Rocks (meat chickens). No morbidity or mortality were observed with doses of 106 or 108EID50/bird when administered by the upper-respiratory route, and the mean infectious dose (106 EID50) was higher than expected, suggesting that the virus is poorly adapted to chickens. Virus was shed at higher titers and spread to the kidneys in chickens inoculated by the intravenous route. Challenge experiments with three other human-origin H7N9 viruses showed a similar pattern of virus replication.

The Impact of Holding Time on the Likelihood of Moving Internally Contaminated Eggs from a Highly Pathogenic Avian Influenza Infected but Undetected Commercial Table-Egg Layer Flock

Emergency response during a highly pathogenic avian influenza (HPAI) outbreak may involve quarantine and movement controls for poultry products such as eggs. However, such disease control measures may disrupt business continuity and impact food security, since egg production facilities often do not have sufficient capacity to store eggs for prolonged periods. We propose the incorporation of a holding time before egg movement in conjunction with targeted active surveillance as a novel approach to move eggs from flocks within a control area with a low likelihood of them being contaminated with HPAI virus. Holding time reduces the likelihood of HPAI-contaminated eggs being moved from a farm before HPAI infection is detected in the flock. We used a stochastic disease transmission model to estimate the HPAI disease prevalence, disease mortality, and fraction of internally contaminated eggs at various time points postinfection of a commercial table-egg layer flock. The transmission model results were then used in a simulation model of a targeted matrix gene real-time reverse transcriptase (RRT)-PCR testing based surveillance protocol to estimate the time to detection and the number of contaminated eggs moved under different holding times. Our simulation results indicate a significant reduction in the number of internally contaminated eggs moved from an HPAI-infected undetected flock with each additional day of holding time. Incorporation of a holding time and the use of targeted surveillance have been adopted by the U.S. Department of Agriculture in their Draft Secure Egg Supply Plan for movement of egg industry products during an HPAI outbreak.

Impact of Virus Strain Characteristics on Early Detection of Highly Pathogenic Avian Influenza Infection in Commercial Table-Egg Layer Flocks and Implications for Outbreak Control

Early detection of highly pathogenic avian influenza (HPAI) infection in commercial poultry flocks is a critical component of outbreak control. Reducing the time to detect HPAI infection can reduce the risk of disease transmission to other flocks. The timeliness of different types of detection triggers could be dependent on clinical signs that are first observed in a flock, signs that might vary due to HPAI virus strain characteristics. We developed a stochastic disease transmission model to evaluate how transmission characteristics of various HPAI strains might effect the relative importance of increased mortality, drop in egg production, or daily real-time reverse transcriptase (RRT)-PCR testing, toward detecting HPAI infection in a commercial table-egg layer flock. On average, daily RRT-PCR testing resulted in the shortest time to detection (from 3.5 to 6.1 days) depending on the HPAI virus strain and was less variable over a range of transmission parameters compared with other triggers evaluated. Our results indicate that a trigger to detect a drop in egg production would be useful for HPAI virus strains with long infectious periods (6-8 days) and including an egg-drop detection trigger in emergency response plans would lead to earlier and consistent reporting in some cases. We discuss implications for outbreak control and risk of HPAI spread attributed to different HPAI strain characteristics where an increase in mortality or a drop in egg production or both would be among the first clinical signs observed in an infected flock.

Moving-Average Trigger for Early Detection of Rapidly Increasing Mortality in Caged Table-Egg Layers

Rapidly increasing and unexplained mortality in commercial poultry flocks may signal the presence of a highly transmissible and reportable disease. Activation of an infectious-disease surveillance system occurs when a key production parameter, i.e., mortality, changes. Various triggers have been proposed to alert producers when mortality exceeds normal limits for a given production system to enable early detection of such diseases. In this article we demonstrate that a simple moving-average trigger is useful for detecting any disease syndrome in caged table-egg layer flocks that manifests itself as sudden, rapidly increasing mortality. We superimposed HPAI disease mortality output data derived from a disease transmission model and from a naturally occurring HPAI outbreak onto normal mortality data from 12 healthy commercial egg-layer flocks, and compared the performance of 7-day moving-average triggers to previously proposed triggers. The moving-average trigger is more efficient, resulting in fewer false-positive alerts and an earlier time to disease detection. It can be easily calculated by using a computer spreadsheet providing only 7 days of mortality data and can be practically and inexpensively implemented by large commercial poultry integrators. A moving-average trigger can be an active component of a production-based surveillance system.

Moving-Average Trigger for Early Detection of Rapidly Increasing Mortality in Caged Table-Egg Layers

Moving-Average Trigger for Early Detection of Rapidly Increasing Mortality in Caged Table-Egg Layers

An Allelotyping PCR for Identifying &lt;em&gt;Salmonella enterica&lt;/em&gt; serovars Enteritidis, Hadar, Heidelberg, and Typhimurium

Current commercial PCRs tests for identifying Salmonella target genes unique to this genus. However, there are two species, six subspecies, and over 2,500 different Salmonella serovars, and not all are equal in their significance to public health. For example, finding S. enterica subspecies IIIa Arizona on a table egg layer farm is insignificant compared to the isolation of S. enterica subspecies I serovar Enteritidis, the leading cause of salmonellosis linked to the consumption of table eggs. Serovars are identified based on antigenic differences in lipopolysaccharide (LPS)(O antigen) and flagellin (H1 and H2 antigens). These antigenic differences are the outward appearance of the diversity of genes and gene alleles associated with this phenotype. We have developed an allelotyping, multiplex PCR that keys on genetic differences between four major S. enterica subspecies I serovars found in poultry and associated with significant human disease in the US. The PCR primer pairs were targeted to key genes or sequences unique to a specific Salmonella serovar and designed to produce an amplicon with size specific for that gene or allele. Salmonella serovar is assigned to an isolate based on the combination of PCR test results for specific LPS and flagellin gene alleles. The multiplex PCRs described in this article are specific for the detection of S. enterica subspecies I serovars Enteritidis, Hadar, Heidelberg, and Typhimurium. Here we demonstrate how to use the multiplex PCRs to identify serovar for a Salmonella isolate.

An Allelotyping PCR for Identifying &lt;em&gt;Salmonella enterica&lt;/em&gt; serovars Enteritidis, Hadar, Heidelberg, and Typhimurium

Current commercial PCRs tests for identifying Salmonella target genes unique to this genus. However, there are two species, six subspecies, and over 2,500 different Salmonella serovars, and not all are equal in their significance to public health. For example, finding S. enterica subspecies IIIa Arizona on a table egg layer farm is insignificant compared to the isolation of S. enterica subspecies I serovar Enteritidis, the leading cause of salmonellosis linked to the consumption of table eggs. Serovars are identified based on antigenic differences in lipopolysaccharide (LPS)(O antigen) and flagellin (H1 and H2 antigens). These antigenic differences are the outward appearance of the diversity of genes and gene alleles associated with this phenotype.We have developed an allelotyping, multiplex PCR that keys on genetic differences between four major S. enterica subspecies I serovars found in poultry and associated with significant human disease in the US. The PCR primer pairs were targeted to key genes or sequences unique to a specific Salmonella serovar and designed to produce an amplicon with size specific for that gene or allele. Salmonella serovar is assigned to an isolate based on the combination of PCR test results for specific LPS and flagellin gene alleles. The multiplex PCRs described in this article are specific for the detection of S. enterica subspecies I serovars Enteritidis, Hadar, Heidelberg, and Typhimurium.Here we demonstrate how to use the multiplex PCRs to identify serovar for a Salmonella isolate.