Detection of antibody to Salmonella enteritidis and S typhimurium in the yolk of hens' eggs

Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect IgG antibodies to Salmonella enteritidis and S typhimurium in the yolk of hens' eggs. Better discrimination and more consistent results were obtained between eggs from experimentally infected and uninfected hens by using saline-dilution of yolk rather than chloroform extraction. Threshold absorbance values were determined in three salmonella-free flocks, and on the basis of these results ELISA optical density values greater than 0.25 were considered to be positive for antibodies to salmonella. Four flocks with a history of salmonella infection were examined; three contained birds which were seropositive for S enteritidis by ELISA and from which S enteritidis was isolated, and a large proportion of eggs from these birds contained antibody to S enteritidis. Eggs from the fourth flock had no detectable antibody, although serum antibody was detected in some birds. No salmonellae were isolated from the yolks of the eggs from any of the four flocks.     

Salmonella enteritidis and other Salmonella in laying hens and eggs from flocks with Salmonella in their environment.

Seven Canadian layer flocks with Salmonella enteritidis in their environment were investigated to determine the numbers of hens infected with S. enteritidis, the localization of S. enteritidis in organs of infected hens and the numbers of S. enteritidis-infected eggs produced by two affected flocks. By a microagglutination test (MAT) using S. pullorum antigens, these flocks had more seropositive hens (mean 51.9 +/- 16.9%) than two Salmonella-free flocks (mean 13.0 +/- 4.2%). Culture of tissues of 580 hens (433 seropositive) from the seven flocks detected 26 (4.5%) S. enteritidis-infected hens from two flocks. In one flock, 2/150 hens were infected with S. enteritidis phage type (PT) 8, which was confined to the ceca, and no Salmonella spp. were isolated from 2520 eggs (one day's lay). In the second flock, where 24/150 hens were infected with S. enteritidis PT13, extraintestinal infection was found in nine hens and involved the ovaries and/or oviduct in two hens. Salmonella enteritidis PT13 was isolated from one sample of egg contents and from one sample of cracked shells from among 14,040 eggs (one day's lay) from this flock. The overall prevalence of S. enteritidis-contaminated eggs from the two flocks with infected hens was less than 0.06%. Other Salmonella spp. isolated were S. heidelberg from 58 hens (10%), and S. hadar, S. mbandaka and S. typhimurium from one hen (0.2%) each. The MAT with antigens of S. pullorum had a sensitivity of 81% and a specificity of 24% for detecting S. enteritidis-infected hens.(ABSTRACT TRUNCATED AT 250 WORDS)     

Development and application of an ELISA for detecting antibodies to Salmonella enteritidis in chicken flocks.

Enzyme-linked immunosorbent assays (ELISAs) were developed for the detection of IgG antibody to Salmonella enteritidis in poultry flocks. A lipopolysaccharide (LPS) and heat-extracted (HE) antigen for use in the ELISA were evaluated together with the rapid slide test (RST), microagglutination test (MT) and the microantiglobulin (MAG) test. In experimentally infected specific pathogen free chickens, good correlation was seen between all tests although, generally, the MT and MAG detected antibody earlier and titres peaked earlier than the ELISAs. The LPS antigen detected antibody earlier than the HE antigen but the latter gave higher titres in the later stages of infection. Cross reactions were seen between S enteritidis and S typhimurium in the ELISAs although homologous reactions were always much higher. Antisera to S montevideo or S senftenberg gave weak positive reactions in both S enteritidis ELISAs. Serological and bacteriological examinations of representative samples from two commercial chicken flocks were carried out. In flock A the HE-ELISA and MAG test detected antibody in nearly all birds. The LPS-ELISA detected antibody in over 60 per cent of birds, while the MT and RST detected few seropositive birds. The whole blood test using the stained S pullorum antigen on the farm detected antibody in just under 25 per cent of the birds. S enteritidis was isolated from the organs of 25 per cent of the birds. All birds in flock B were seronegative by all tests; no salmonellae were isolated from the organs of these birds.     

Vaccination against Salmonella enteritidis in Dutch commercial layer flocks with a vaccine based on a live Salmonella gallinarum 9R strain: evaluation of efficacy, safety, and performance of serologic Salmonella tests

This study describes a field trial in which 80 commercial layer flocks, with an increased risk of Salmonella enteritidis (SE) infection and placed on farms with a certified Standardized Biosecurity Programme (SBP) or a request for a SBP certificate, were vaccinated with a vaccine based on a live attenuated Salmonella gallinarum (SG) 9R strain. An evaluation is presented of the efficacy of the vaccine against SE infections, the effect on the performance of serologic Salmonella tests, and the spread of the vaccine strain to the egg content. For the efficacy study, assessment of the flock level occurrence of SE infections in the vaccinated group of 80 flocks was compared with that of a nonvaccinated group of 1854 flocks hatched in the same period. This control group was examined according to the compulsory control programme in The Netherlands. An evaluation was done of the performance of serologic Salmonella tests and the spread of the vaccine strain to the inner egg content of five of the vaccinated flocks. Findings demonstrated the flock level occurrence of SE infections in the vaccinated group (2/80 = 2.5%) to be significantly (P = 0.01) lower than that of the nonvaccinated group (214/1854 = 11.5%). Vaccination resulted in 59.0% positive test results in lipopolysaccharide BD enzyme-linked immunosorbent assay (ELISA) for detecting antibodies against Salmonella serogroups B and D and 0% positive test results in the rapid plate agglutination test for detecting antibodies against S. pullorum (SP)/SG. The mean specificities of two blocking ELISAs (gm- and i-double antibody sandwich ELISAs) based on the flagellar antigen of SE and Salmonella typhimurium (ST) on the same sera were 99.6% and 96.1%, respectively. The vaccine strain could not be isolated from any of the 450 pools of 10 eggs. On the basis of these results, we concluded that vaccination with a vaccine based on an attenuated SG 9R strain contributes to the reduction of SE infections in commercial layer flocks. Furthermore, serologic monitoring of SE, ST, and SP/SG can still be carried out on flocks vaccinated with an attenuated SG 9R strain. Additionally, we found no indication of the spread of the vaccine strain to the egg content.     

Isolation and characterization of a 6/85-like Mycoplasma gallisepticum from commercial laying hens

Eighty-three-week-old table egg layers with swollen sinuses were presented with a history of increased mortality. Serology revealed positive titers to Mycoplasma gallisepticum (MG). The birds were part of a flock in which some birds had been vaccinated with 6/85 live MG vaccine at 18 wk of age. Tracheal cultures were obtained from both vaccinated and unvaccinated birds within the flock. The cultures were indistinguishable from 6/85 vaccine by both random amplified polymorphic DNA analysis and DNA sequence analysis. Challenge studies were performed to compare the field isolates with 6/85 vaccine and the R strain of MG. The field isolates produced a greater antibody response by serum plate agglutination than did the 6/85 vaccine. The isolates effectively colonized the trachea without increasing the tracheal mucosal thickness; however, they did not extensively colonize the air sacs or cause airsacculitis in the experimental birds.     

Salmonella contamination of poultry flocks in The Netherlands

The contamination of poultry in the Netherlands with Salmonella enteritidis was tested. For this, different methods (detection of S. enteritidis in faecal samples of 25 g; detection of S. enteritidis in cloacal swabs; detection of S. enteritidis by serological testing of antibodies in serum) were compared for their efficiency to detect S. enteritidis in flocks of poultry. Testing of faecal samples clearly yielded the best results. This method was used in a transmission study, in which 14 flocks descending from a contaminated primary mother flock were screened for the presence of S. enteritidis. The method was also used for screening 49 flocks of laying hens and 52 flocks of broiler chickens throughout the Netherlands. From the transmission study it became clear that S. enteritidis, phage type 2 (Dutch phage set) was isolated both from the mother flock and from five of the descendent flocks. Screening of poultry flocks for the presence of salmonella revealed that salmonella was present in 47% of the layer flocks and in 94% of the broiler flocks. S. enteritidis was isolated from 15% of the flocks screened.     

Plasmid analysis and epidemiology of Salmonella enteritidis infection in three commercial layer flocks.

Ninety-six S. enteritidis isolates obtained from three commercial layer flocks in 1988-90 were examined following DNA extraction, restriction enzyme digestion, and gel electrophoresis for plasmid size profiles and restriction fragment length polymorphisms (RFLPs). The S. enteritidis isolates from the three flocks had three, eight, and two different plasmid profiles, respectively. Only four isolates from one flock lacked plasmids. A 36-megadalton (mDa) (54-kilobase) plasmid was present in 73% of the isolates, either alone or in combination with other plasmids. Isolates with only the 36-mDa plasmid had identical RFLPs. The diversity of plasmid profiles was greater than that of phage-types among isolates from the three flocks: 12 unique plasmid profiles vs. four phage-types. Mixed infections with S. enteritidis strains having distinct plasmid profiles occurred in all three flocks. Reinfection of these flocks in 1990 with one or more of the strains obtained earlier was evident, because some of the original isolates and the 1990 isolates had matching plasmid profiles and were of the same phage-types. Isolates from both environmental and tissue samples, examined from one flock, were found to share the same plasmid profile and phage-type.     

Antibody prevalence of low-pathogenicity avian influenza and evaluation of management practices in Minnesota backyard poultry flocks

Low‐pathogenicity avian influenza (LPAI) viruses have caused illness in poultry and humans with poultry contact. To determine whether there is evidence of exposure to avian influenza viruses (AIV) among backyard poultry in Minnesota and their human caretakers, 150 flocks of backyard birds were sampled for antibodies to AIV from August 2007 through December 2008. One hundred flocks were tested through routine slaughter surveillance by the Minnesota Board of Animal Health and an additional 50 flocks were contacted and sampled by study investigators. Blood was collected from 10 to 13 birds from each flock and a survey of biosecurity and management practices was administered to the flock owner. Blood samples were tested by agar gel immunodiffusion (AGID) for influenza A antibodies. Tested flocks had a median flock size of 100 birds (range: 12–800 birds), and were most commonly owned for meat for personal use (81% of respondents), fun or hobby (58%) and eggs for personal use (56%). Although 7% of flock owners reported that their birds had shown respiratory signs in the previous 3 months, only 1 of 150 flocks tested positive for influenza by AGID. Antibodies to LPAI H6N1 were detected in the positive flock. The owner of the positive flock did not have antibodies to H6 or other common AIV. Based on the findings of this study, the risk of transmission of LPAI viruses from backyard poultry to owners in Minnesota appears to be low under current conditions and management practices.     

Evaluation of the efficacy of Salmonella enteritidis oil-emulsion bacterin in an intravaginal challenge model in hens.

The efficacy of Salmonella enteritidis (SE) oil-emulsion bacterin (a commercially available vaccine) was evaluated in an intravaginal challenge model in hens producing a high rate of SE-contaminated eggs. Hens were vaccinated at 38 wk of age. A second (booster) bacterin injection was administered 4 wk later. Two weeks after the second vaccination, all hens were challenged intravaginally with 10(7) colony-forming units of SE. After challenge, 36 of 189 eggs (19.0%) in the vaccinated hens were positive for SE, and this contamination rate was significantly (P < 0.01) lower than that in the unvaccinated hens (61 of 165 eggs, 37.0%). SE was highly recovered from the cloacal and vaginal swabs of the unvaccinated and vaccinated hens, but the number of SE from the cloaca of the vaccinated hens was significantly (P < 0.05) lower than that in the unvaccinated hens at 7 days post-challenge (PC). The recoveries of SE from the spleen and ovary in the vaccinated hens were significantly (P < 0.05) lower than those in the unvaccinated hens at 7 days PC. At necropsy, SE was recovered from 2 of 15 forming eggs (13.3%) taken from the oviducts of the unvaccinated hens, whereas no SE was recovered from 17 forming eggs in the vaccinated hens. After vaccination, serum antibodies for SE in the vaccinated hens were significantly higher than those in the unvaccinated hens. Antibodies from the oviductal washing, especially immunoglobulin G isotype, in the vaccinated hens were higher than those in the unvaccinated hens after challenge. This intravaginal challenge model produced frequent contaminated eggs and clearly demonstrated the ability of the bacterin to protect against egg contamination. The present model may be a useful tool for further studies to evaluate the protective effect against SE contamination of eggs by potential vaccine candidates.     

Infection dynamics of Ascaridia galli in non-caged laying hens

The infection dynamics of Ascaridia galli in laying hens was investigated in six commercial non-caged flocks. Three flocks were managed in accordance with the regulations for organic production and had outdoor access, whereas three flocks were housed indoors in aviaries or traditional floor systems. Faecal egg counts and total worm burdens were determined at specified intervals during the first 50 weeks of the production period. In two conventional flocks the efficacy of flubendazole on lumenal stages was investigated. All flocks became infected following the arrival of the birds (post placement) with residual infective eggs derived from the previous flock. In four flocks (two organic and two conventional) parasite eggs were first detected in faeces 6-7 weeks post placement, whereas parasite eggs were not detected until after 17-18 weeks in two flocks. This delay was observed in two of three flocks that were housed in barns that had been thoroughly cleaned and disinfected by chlorocresol. In three flocks (two conventional and one organic) flubendazole was administered to the birds in the drinking water for approximately one week. Both conventional flocks were dewormed twice approximately 20 weeks apart, whereas the organic flock was dewormed only once about 40 weeks post placement. Parasite eggs reappeared after deworming in all flocks, often within 2-4 weeks, followed by a rapid increase in parasite egg expulsion. Our results suggested impairment of host immunity post treatment, as the egg counts exceeded pre-treatment levels after 7-8 weeks on both conventional farms. Accordingly, the way by which anthelmintics and/or disinfectants are used in non-caged chicken flocks must be refined.     

Economic effects of clinical chicken anemia agent infection on profitable broiler production.

An outbreak of anemia dermatitis syndrome caused by chicken anemia agent (CAA) occurred in 15 broiler flocks. An average of 29% of chickens in these flocks were derived from a common breeder flock. The breeder flock had no antibody to CAA at 20 weeks of age but had seroconverted by 31 weeks. Diseased broiler flocks were derived from eggs laid by the breeder flock between 25 and 30 weeks of age. CAA infection in the breeder flock was subclinical, with no apparent effects on mortality or performance. A strategic program of therapeutic and/or prophylactic antibiotic therapy was begun in affected broiler flocks as soon as the disease was diagnosed. Nevertheless, when the cost of therapy was taken into account, affected broiler flocks had a net income 17.3% to 19.6% lower than normal flocks. Average bird weights were 3.3% to 3.5% lower in affected flocks than in unaffected flocks, and affected flocks had a significantly greater proportion of lighter birds. Average mortality in affected flocks was 2.0% to 2.3% higher than in normal flocks, with peak mortality occurring in the third week of life. There was no apparent effect on feed-conversion ratio.     

Results of a Salmonella enteritidis vaccination field trial in broiler-breeder flocks in The Netherlands

From August 1995 until December 1997, the effect of adding Salmonella enteritidis (SE) vaccination to a certified standardized biosecurity program in a situation of increased infection risk was examined in a field trial in The Netherlands. In this field trial, two groups of broiler-breeder flocks with increased infection risk were vaccinated, one group with VAC-T/TALOVAC logSE(group A) and the second group with SALENVAC (group B). The determination of increased infection risk in groups A and B was based on an SE infection history; flocks were either previously infected and treated (PIT) or had other risk factors than previously infected and treated (OPIT). SE infections in both vaccinated groups were assessed by monitoring according to the Dutch salmonella control program. Under field conditions, designation of a vaccinated and a control group on the farm was not possible. In the same period as the vaccinated groups, 608 nonvaccinated flocks (group C) were hatched and monitored according to the Dutch salmonella control program. The flock level occurrence of SE infection in the vaccinated groups was compared with the flock level occurrence of SE infection in the nonvaccinated group on the basis of comparability of infection risk. In group C, whether or not flocks had infection risk PIT was known and for risk factor OPIT, only whether or not a flock had been placed on a previously contaminated farm (= risk of reinfection) was known. The proportion of SE-infected flocks with risk factor PIT in the vaccinated groups was not significantly different from that in the nonvaccinated group C. Only the proportion of SE-infected flocks with a risk of reinfection in the vaccinated group B (0) was significantly lower (P = 0.02) than in the nonvaccinated group C (18%). The fact that no significant result was found in favor of group A is because of the small number of flocks in this part of the study. On the basis of the conditions of the setup of this trial, it can only be concluded that there is an indication that vaccination contributes in the reduction of SE reinfection in broiler breeder flocks.     

The effect of killed Salmonella enteritidis vaccine prior to induced molting on the shedding of s. enteritidis in laying hens

Effects of administering killed Salmonella enterica serovar enteritidis (SE) vaccines to laying hens prior to induced molting on egg production and on shedding of SE were investigated. Forty hens were vaccinated with one of two SE vaccines available commercially in the United States and Japan. Twenty-five days after vaccination, feed was withdrawn for 2 wk from 20 vaccinated plus 10 unvaccinated hens to induce molt. Four days after molt induction, all hens were challenged with a dose of 2.4 X 10(9) of SE. For the 25 days following administration of the SE bacterins, egg production in vaccinated hens showed approximately a 15% decrease. After molt induction, egg production in molted hens ceased and then returned to normal levels 8 or 9 wk postvaccination. Through the 3-mo experimental period, the decreases in numbers of eggs laid in the unvaccinated/molted group and two vaccinated/molted groups were 225 (26.2%), 245 (28.4%), and 274 (31.9%), respectively, compared with 860 in the unvaccinated/unmolted group. There was no significant difference in egg lay at the P < 0.05 level among the former three groups. Hens in the vaccinated/molted groups shed about two logs less SE than hens in the unvaccinated/molted group 3 14 days postchallenge (P < 0.05 or 0.01). These results indicate that vaccination prior to induced molting might be effective in preventing the exacerbation of SE problems within flocks in which the potential for SE contamination may exist.     

Epidemiologic characterization of Colorado backyard bird flocks

Backyard gallinaceous bird flocks may play an important role in the spread of infectious diseases within poultry populations as well as the transmission of zoonotic diseases to humans. An epidemiologic characterization was conducted of Colorado backyard flocks to gather information on general flock characteristics, human movement of birds, human-bird interaction, biosecurity practices, and flock health. Our results suggest that backyard poultry flocks in Colorado are small-sized flocks (68.6% of flocks had < 50 birds); consist primarily of layer chickens (85.49% of flocks), show chickens (32.18% of flocks), and waterfowl (34.07% of flocks); and are primarily owned for food (meat or egg) production for the family (86.44%) or as pet or hobby birds (42.27%). The backyard flock environment may promote bird-to-bird transmission as well as bird-to-human transmission of infectious disease. Birds are primarily housed with free access to the outside (96.85%), and many are moved from the home premises (46.06% within 1 yr). Human contact with backyard flocks is high, biosecurity practices are minimal, and bird health is negatively impacted by increased movement events. Increased knowledge of backyard bird characteristics and associated management practices can provide guidelines for the development of measures to decrease disease transmission between bird populations, decrease disease transmission from birds to humans, and increase the overall health of backyard birds.     

Monitoring the Immune Status of Broilers Against Reoviruses Using Challenge and Serologic Data

Reoviruses are an important cause of suboptimum performance in commercial broilers worldwide. Integrators use the enzyme-linked immunosorbent assay against the S1133 antigen for monitoring serum of breeders for indicating pullet vaccine success. However, without correlating serology to reovirus challenge, it is difficult to determine whether titers reflect protective immunity. We developed a broiler challenge test against 2 common reovirus isolates (2408 and S1133) to evaluate the efficacy of reovirus pullet vaccine programs. Two reovirus serologic and challenge studies were undertaken using chicks from broiler integrators from the southeastern United States. Breeder flocks, from which the chicks were obtained, received at least 1 live and 2 inactivated reovirus vaccines during their pullet phase. One-day-old progeny were collected from 6 breeder flocks. At 1 d of age, 20 chicks from each broiler flock were bled, and serum was analyzed for antibodies. At 3 to 4 d of age, 20 progeny per flock were challenged with the 2408 reovirus by intratracheal route. At 10 to 14 d of age, another 20 birds per flock were challenged with the S1133 reovirus by footpad. Twenty birds per flock were used as nonchallenged controls. At 3 wk of age, all birds were killed and weighed. Percentage of protection was calculated for each flock based on the absence of gross lesions. Flocks with at least 50% protection were considered well protected. Most flocks were well protected against both viruses. The percentage of protection correlated with day-old enzyme-linked immunosorbent assay titers. Chicks from younger hens had higher titers and the best protection against challenge. Producers, whose hen flocks were monitored herein, were doing a good job of immunizing pullets against reovirus. They are now using reovirus progeny challenge studies along with breeder antibody titers to determine vaccination success of their pullets.     

Characteristics of two duck farming systems in the Mekong Delta of Viet Nam: stationary flocks and moving flocks, and their potential relevance to the spread of highly pathogenic avian influenza

Ducks are considered to play a major role in the spread of highly pathogenic avian influenza (HPAI) H5N1 in Viet Nam, but detailed information on their management is limited. We distinguished two different systems (1) stationary duck flocks that are not commonly driven to rice fields beyond village boundaries and that are confined overnight on farms and (2) moving duck flocks that are intentionally driven to rice fields beyond village boundaries, that are not returning to home farms for extended periods and that are housed overnight in temporary enclosures in rice paddies. A total of 115 stationary and 22 moving flock farmers were interviewed in 2007 in the Mekong Delta of Viet Nam. Moving duck flocks are larger than stationary flocks, which is indicative of their more commercial production. Moving flock farmers apparently are more aware of HPAI risks than stationary flock farmers, as their flocks are more likely fully vaccinated and have less contact with chickens during scavenging. On the other hand, the spread of HPAI virus between birds might be promoted by moving duck flocks as they repeatedly use transport vehicles and numerous rice paddies for scavenging and are often visited by hatchery owners in the field for purchasing duck eggs. In addition, long distances travelled by moving duck flocks might also result in widespread dissemination of HPAI virus. Further studies are necessary to describe HPAI prevalence and travel patterns of moving duck flocks and to explore the moving duck flock network in detail.     

Effects of vaccination and other preventive methods for Salmonella enteritidis on commercial laying chicken farms

The effect of introducing vaccinated commercial laying chickens on to farms, which previously had laying flocks that were infected with Salmonella Enteritidis, was investigated by sampling faeces and environmental samples, and in some cases spent hens. In 15 of 17 free-range flocks vaccination eliminated any evidence of infection. In 11 barn egg production flocks, vaccination produced similar results in four flocks on one farm but infection persisted in seven flocks on other farms. Vaccination of two consecutive cage layer flocks led to a gradual disappearance of the infection, but in 18 other flocks there was evidence of infection after vaccination. In one continuously occupied cage layer house, treatment by competitive exclusion was followed by a gradual disappearance of S Enteritidis in faeces and a substantial reduction in its levels in the environment. On four barn egg production sites disinfection with a formaldehyde, glutaraldehyde and quaternary ammonium compound disinfectant eliminated Salmonella species even though birds housed subsequently were not vaccinated. In three flocks that had been vaccinated for four years, S Enteritidis was still present. In most cases the poor performance of the vaccine was associated with severe rodent control problems and a poor standard of cleaning and disinfection.     

Phenotypic and genotypic characterization of Salmonella arizonae from an integrated turkey operation

Fifty cases submitted between 2000 and 2002 were selected for retrospective analysis to evaluate possible relationships between Salmonella arizonae isolated from breeder flocks, hatching eggs, and meat bird flocks belonging to a single turkey integrator. In all the meat bird cases selected for this study, arizonosis was the primary diagnosis. In birds under 1 month of age, clinical signs and pathologic changes were observed in older birds. The Salmonella arizonae isolates were analyzed by antibiotic resistance pattern and serotype and genotyped by pulsed-field gel electrophoresis (PFGE). Serotyping and PFGE yielded similar results, but the antibiotic resistance patterns did not correspond to either serotyping or PFGE typing. The presence of common pulsed-field patterns in breeder flocks, eggs, and meat bird flocks suggested that S. arizonae was being transmitted vertically from the breeder flock.     

Sero-prevalence of avian influenza among broiler-breeder flocks in Jordan

Thirty blood samples were collected randomly from each of the 38 breeder-broiler farms in Jordan. Serum samples were examined using indirect ELISA for specific antibodies to avian influenza virus. The overall true flock-level sero-prevalence of avian influenza was 71% (95% CI: 55,83). Positive flocks had 2-30 sero-positive chickens and half of flocks had >20 sero-positive birds. The number of sero-positive flocks varied in the studied localities with more sero-positives in farms located within the migratory route of migratory wild fowl. The examined broiler-breeder flocks had no clinical signs, or noticeable decrease in egg production; mortalities were within the normal range (0.1-1%). The number of positive sera/flock correlated with flock size. There were a no significant (Pearsons r=0.21, p=0.21) correlation between positive flocks and age. A non-pathogenic AI virus infects broiler-breeder farms in Jordan. Wild local and migrating birds might promote the further spread of this virus in Jordan and other countries.