Turkey adenovirus 3, a siadenovirus, uses sialic acid on N-linked glycoproteins as a cellular receptor.

Turkey adenovirus 3 (TAdV-3) is the causative agent of an immune-mediated disease in turkeys, haemorrhagic enteritis, through targeting B lymphocytes. In the present study, we investigated the role of sialic acid in TAdV-3 entry and characterized the structural components of TAdV-3 receptor(s) on RP19, B lymphoblastoid cells. Removal of the cell-surface sialic acids by neuraminidases or blocking of sialic acids by wheat germ agglutinin lectin reduced virus infection. Pre-incubation of cells with Maackia amurensis lectin or Sambucus nigra agglutinin resulted in virus reduction, suggesting that TAdV-3 uses both α2,3-linked and α2,6-linked sialic acids as attachment receptor. Virus infectivity data from RP19 cells treated with sodium periodate, proteases (trypsin or bromelain) or metabolic inhibitors (dl-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol, tunicamycin, or benzyl N-acetyl-α-d-galactosaminide) indicated that N-linked, but not O-linked, carbohydrates are part of the sialylated receptor and they are likely based on a membrane glycoprotein, rather than a glycolipid. Furthermore, our data, in conjunction with previous findings, implies that the secondary receptor for TAdV-3 is a protein molecule since the inhibition of glycolipid biosynthesis did not affect the virus infection, which was rather reduced by protease treatment. We can conclude that terminal sialic acids attached to N-linked membrane glycoproteins on B cells are used for virus attachment and are essential for successful virus infection.

In vitro growth kinetics and gene expression analysis of the turkey adenovirus 3, a siadenovirus.

Turkey adenovirus 3 (TAdV-3) belongs to the genus Siadenovirus, family Adenoviridae. Previously, nucleotide sequencing and annotation of the Virginia avirulent strain (VAS) of TAdV-3 genome, isolated in our laboratory, indicated the presence of a total of 23 genes and open reading frames (ORFs). The goals of this study were 1) to delineate the growth kinetics of the virus using a qPCR-based infectivity assay, and 2) to determine the virus gene expression profile during the early and late phases of infection in target B lymphocytes. The one-step growth curve experiment demonstrated 3 phases of virus replication cycle: a lag phase lasted for 12-18 h post-infection (h.p.i.), in which the virus titer declined; a log phase from 18 to 120 h.p.i., in which the number of infectious virus particles increased over 20,000 folds, and a brief decline phase thereafter. Southern blot analysis indicated that the synthesis of new viral DNA started by 8 h.p.i. Gene-specific RT-PCR analysis revealed the expression of mRNAs from the 23 TAdV-3 genes/ORFs. According to the temporal transcriptional profiling of TAdV-3 genome, genes could be divided into 3 groups based on the time of transcription initiation: group 1 showed detectable levels of transcription at 2 h.p.i and included 7 genes, i.e., hyd, III, pX, pVI, II, 100 K, and 33 K; group 2 included 12 genes whose mRNAs were detected for the first time at 4 h.p.i., i.e., ORF1, IVa2, pol, pTP, pIIIa, EP, DBP, E3, U exon, IV, ORF7, and ORF8; group 3 of transcripts were detectable starting 8 h.p.i. and included only 4 genes, i.e., 52 K, 22 K, pVII, and pVIII. Our data suggest that the transcriptional kinetics of genus Siadenovirus differ from that observed in other adenoviral genera; however, a few TAdV-3 genes showed similar expression patterns to their adenoviral homologs.

Effect of Different Instructional Methods on Contamination and Personal Protective Equipment Protocol Adherence among Veterinary Students.

Proper use of personal protective equipment (PPE) is crucial to prevent disease spread. Recent studies in human medicine have shown disconcerting inconsistencies in the use of PPE in hospital wards. In this study, we compared the effect of three instructional methods for PPE use on contamination and protocol adherence among veterinary students. Students were divided into three groups according to the instructional method to which they had access (instructional video, wall chart, or both). They underwent an isolation exercise consisting of donning, patient examination (mock patient prepared with contamination marker), and doffing. Student contamination after the exercise was evaluated using UV light. Videos of student performance were reviewed for errors committed. Results showed that the number of students with contamination was higher in the group who only had access to video instruction than in the two other groups. The number of students with contamination on forearms, hands, and wrists was higher in the group who only had access to charts. Disinfecting gloves between doffing steps was the most frequently omitted step. The number of students who touched the environment with unprotected areas of their bodies was higher in the group who only had access to video instruction than in the other two groups. In conclusion, video instruction was less effective in achieving PPE protocol adherence among veterinary students than was instruction with a chart or chart-video combination. Incorporating video instruction as part of the instructions may be valuable to reinforce individual steps of donning and doffing.

Biofilm formation and avian immune response following experimental acute and chronic avian cholera due to Pasteurella multocida.

Pasteurella multocida is the causative agent of avian cholera, an important economic and ecological disease that can present as a peracute, acute, chronic, or asymptomatic infection. Acute avian cholera is associated with encapsulated P. multocida, while chronic and asymptomatic cases of avian cholera may be associated with capsule-deficient P. multocida isolates. We hypothesize that biofilm formation is also associated with chronic and asymptomatic avian cholera. Experimental infections of chickens with encapsulated, biofilm-deficient P. multocida strain X73, proficient biofilm forming P. multocida strain X73ΔhyaD, and proficient biofilm forming clinical strains 775 and 756 showed that virulence was inversely correlated with biofilm formation. Biofilm-proficient isolates induced chronic avian cholera in the chicken host. Histopathological analysis was used to show that biofilm-proficient isolates induced little inflammation in the lungs, heart, and liver, while biofilm-deficient isolates induced greater inflammation and induced the recruitment of heterophil granulocytes. Putative biofilm matrix material and exopolysaccharide was detected in pulmonary tissue of chickens diagnosed with chronic avian cholera using scanning electron microscopy and a fluorescently-tagged lectin, respectively, supporting a role for biofilm in chronic avian cholera. P. multocida induced Th1 and Th17 immune responses during acute and chronic avian cholera, as determined by quantitative real-time PCR of splenic cytokine genes. Chickens that succumbed to acute avian cholera after experimental challenge with strain X73 had high levels of INF-γ, IL-1ß, IL-6, IL-12A, IL-22, IL-17A, and IL-17RA expressed in the spleen compared to all other experimental groups. Birds infected with capsule-deficient strains had chronic infections lasting 7 days or longer, and had increased levels of IL-17RA, CCR6, and IL-16 compared to non-infected control chickens. However, specific antibody titers increased only transiently to capsule-deficient strains and were low, indicating that antibodies are less important in managing and clearing P. multocida infections.

Review of Nonfoodborne Zoonotic and Potentially Zoonotic Poultry Diseases.

Emerging and re-emerging diseases are continuously diagnosed in poultry species. A few of these diseases are known to cross the species barrier, thus posing a public health risk and an economic burden. We identified and synthesized global evidence for poultry nonfoodborne zoonoses to better understand these diseases in people who were exposed to different poultry-related characteristics (e.g., occupational or nonoccupational, operational types, poultry species, outbreak conditions, health status of flocks). This review builds on current knowledge on poultry zoonoses/potentially zoonotic agents transmitted via the nonfoodborne route. It also identifies research gaps and potential intervention points within the poultry industry to reduce zoonotic transmission by using various knowledge synthesis tools such as systematic review (SR) and qualitative (descriptive) and quantitative synthesis methods (i.e., meta-analysis). Overall, 1663 abstracts were screened and 156 relevant articles were selected for further review. Full articles (in English) were retrieved and critically appraised using routine SR methods. In total, eight known zoonotic diseases were reviewed: avian influenza (AI) virus (n = 85 articles), Newcastle disease virus (n = 8), West Nile virus (WNV, n = 2), avian Chlamydia (n = 24), Erysipelothrix rhusiopathiae (n = 3), methicillin-resistant Staphylococcus aureus (MRSA, n = 15), Ornithonyssus sylvarium (n = 4), and Microsporum gallinae (n = 3). In addition, articles on other viral poultry pathogens (n = 5) and poultry respiratory allergens derived from mites and fungi (n = 7) were reviewed. The level of investigations (e.g., exposure history, risk factor, clinical disease in epidemiologically linked poultry, molecular studies) to establish zoonotic linkages varied across disease agents and across studies. Based on the multiple outcome measures captured in this review, AI virus seems to be the poultry zoonotic pathogen that may have considerable and significant public health consequences; however, epidemiologic reports have only documented severe human cases clustered in Asia and not in North America. In contrast, avian Chlamydia and MRSA reports clustered mainly in Europe and less so in North America and other regions. Knowledge gaps in other zoonoses or other agents were identified, including potential direct (i.e., nonmosquito-borne) transmission of WNV from flocks to poultry workers, the public health and clinical significance of poultry-derived (livestock-associated) MRSA, the zoonotic significance of other viruses, and the role of poultry allergens in the pathophysiology of respiratory diseases of poultry workers. Across all pathogens reviewed, the use of personal protective equipment was commonly cited as the most important preventive measure to reduce the zoonotic spread of these diseases and the use of biosecurity measures to reduce horizontal transmission in flock populations. The studies also emphasized the need for flock monitoring and an integrated approach to prevention (i.e., veterinary-public health coordination with regard to diagnosis, and knowledge translation and education in the general population) to reduce zoonotic transmission.

Preharvest Salmonella Detection for Evaluation of Fresh Ground Poultry Product Contamination.

Salmonella is an important economic and public health concern for the poultry industry. Fresh ground product has been linked with multiple salmonellosis outbreaks in humans. Exposure can be controlled by proper handling and preparation by consumers; however, the industry desires to minimize carriage levels in the final product. A substantial obstacle in reducing product contamination stems from limitations in diagnostic methodologies. Detection of Salmonella contamination currently requires extended incubation periods, and by the time test results are available, the fresh product has reached retail shelves. The goal of this study was to develop a preharvest diagnostic protocol for the evaluation of ground product contamination. The turkey processing plant where this research was conducted had previously established Salmonella screening (BAX system) of ground product, thus providing an opportunity for preharvest sample comparison. Drag swabs were collected from live-haul trailers entering the processing plant over a 12-month period. The swabs were added to modified buffered peptone water and incubated at 40°C. After incubation for 6 h or overnight, samples were tested for the presence of Salmonella with the DNAble assay and related to ground turkey samples from corresponding lots. The linear relationship for the percentage of Salmonella-positive live-haul trailers was significant for both the 6-h (slope = 1.02, R(2) = 0.96, and P < 0.0001) and overnight (slope = 0.35, R(2) = 0.93, and P = 0.0015) incubations, with the percentage of Salmonella-positive ground turkey samples. These data indicate that preharvest screening provides a meaningful evaluation of product contamination. Additionally, the 6-h incubation protocol is rapid enough to allow for product mitigation and could potentially aid in the reduction of future salmonellosis outbreaks.

RNA transcripts of full-length cDNA clones of rabbit hepatitis E virus are infectious in rabbits.

Hepatitis E virus (HEV), the causative agent of hepatitis E, is a single-stranded positive-sense RNA virus belonging to the family Hepeviridae. At least four genotypes of the family infect humans: genotypes 1 and 2 are transmitted to humans through contaminated water, while genotypes 3 and 4 are zoonotic and have animal reservoirs. A novel strain of HEV recently identified in rabbits is a distant member of genotype 3, and thus poses a potential risk of zoonotic transmission to humans. The objective of this study was to construct and characterize an infectious cDNA clone of the rabbit HEV. Two full-length cDNA clones of rabbit HEV, pT7g-rabHEV and pT7-rabHEV, were constructed and their infectivity was tested by in vitro transfection of Huh7 human liver cells and by direct intrahepatic inoculation of rabbits with capped RNA transcripts. Results showed that positive signal for rabbit HEV protein was detected by an immunofluorescence assay with a HEV-specific antibody in Huh7 human liver cells transfected with capped RNA transcripts from the two full-length cDNA clones. Rabbits intrahepatically inoculated with capped RNA transcripts from each of the two clones developed active HEV infection as evidenced by seroconversion to anti-HEV antibodies, and detection of rabbit HEV RNA in sera and feces of inoculated animals. The availability of a rabbit HEV infectious cDNA clone now affords us the ability to delineate the mechanism of HEV replication and cross-species infection in a small animal model.

The PSAP motif within the ORF3 protein of an avian strain of the hepatitis E virus is not critical for viral infectivity in vivo but plays a role in virus release.

The ORF3 protein of hepatitis E virus (HEV) is a multifunctional protein important for virus replication. The ORF3 proteins from human, swine, and avian strains of HEV contain a conserved PXXP amino acid motif, resembling either Src homology 3 (SH3) cell signaling interaction motifs or "late domains" involved in host cell interactions aiding in particle release. Using an avian strain of HEV, we determined the roles of the conserved prolines within the PREPSAPP motif in HEV replication and infectivity in Leghorn male hepatoma (LMH) chicken liver cells and in chickens. Each proline was changed to alanine to produce 8 avian HEV mutants containing single mutations (P64, P67, P70, and P71 to A), double mutations (P64/67A, P64/70A, and P67/70A), and triple mutations (P64/67/70A). The results showed that avian HEV mutants are replication competent in vitro, and none of the prolines in the PXXPXXPP motif are essential for infectivity in vivo; however, the second and third prolines appear to aid in fecal virus shedding, suggesting that the PSAP motif, but not the PREP motif, is involved in virus release. We also showed that the PSAP motif interacts with the host protein tumor suppressor gene 101 (TSG101) and that altering any proline within the PSAP motif disrupts this interaction. However, we showed that the ORF2 protein expressed in LMH cells is efficiently released from the cells in the absence of ORF3 and that coexpression of ORF2 and ORF3 did not act synergistically in this release, suggesting that another factor(s) such as ORF1 or viral genomic RNA may be necessary for proper particle release.

Mutational analysis of the hypervariable region of hepatitis e virus reveals its involvement in the efficiency of viral RNA replication.

The RNA genome of the hepatitis E virus (HEV) contains a hypervariable region (HVR) in ORF1 that tolerates small deletions with respect to infectivity. To further investigate the role of the HVR in HEV replication, we constructed a panel of mutants with overlapping deletions in the N-terminal, central, and C-terminal regions of the HVR by using a genotype 1 human HEV luciferase replicon and analyzed the effects of deletions on viral RNA replication in Huh7 cells. We found that the replication levels of the HVR deletion mutants were markedly reduced in Huh7 cells, suggesting a role of the HVR in viral replication efficiency. To further verify the results, we constructed HVR deletion mutants by using a genetically divergent, nonmammalian avian HEV, and similar effects on viral replication efficiency were observed when the avian HEV mutants were tested in LMH cells. Furthermore, the impact of complete HVR deletion on virus infectivity was tested in chickens, using an avian HEV mutant with a complete HVR deletion. Although the deletion mutant was still replication competent in LMH cells, the complete HVR deletion resulted in a loss of avian HEV infectivity in chickens. Since the HVR exhibits extensive variations in sequence and length among different HEV genotypes, we further examined the interchangeability of HVRs and demonstrated that HVR sequences are functionally exchangeable between HEV genotypes with regard to viral replication and infectivity in vitro, although genotype-specific HVR differences in replication efficiency were observed. The results showed that although the HVR tolerates small deletions with regard to infectivity, it may interact with viral and host factors to modulate the efficiency of HEV replication.

Construction of an infectious cDNA clone of avian hepatitis E virus (avian HEV) recovered from a clinically healthy chicken in the United States and characterization of its pathogenicity in specific-pathogen-free chickens.

A genetically distinct strain of avian hepatitis E virus (avian HEV-VA strain) was isolated from a healthy chicken in Virginia, and thus it is important to characterize and compare its pathogenicity with the prototype strain (avian HEV-prototype) isolated from a diseased chicken. Here we first constructed an infectious clone of the avian HEV-VA strain. Capped RNA transcripts from the avian HEV-VA clone were replication-competent after transfection of LMH chicken liver cells. Chickens inoculated intrahepatically with RNA transcripts of avian HEV-VA clone developed active infection as evidenced by fecal virus shedding, viremia, and seroconversion. To characterize the pathogenicity, RNA transcripts of both avian HEV-VA and avian HEV-prototype clones were intrahepatically inoculated into the livers of chickens. Avian HEV RNA was detected in feces, serum and bile samples from 10/10 avian HEV-VA-inoculated and 9/9 avian HEV-prototype-inoculated chickens although seroconversion occurred only in some chickens during the experimental period. The histopathological lesion scores were lower for avian HEV-VA group than avian HEV-prototype group in the liver at 3 and 5 weeks post-inoculation (wpi) and in the spleen at 3 wpi, although the differences were not statistically significant. The liver/body weight ratio, indicative of liver enlargement, of both avian HEV-VA and avian HEV-prototype groups were significantly higher than that of the control group at 5 wpi. Overall, the avian HEV-VA strain still induces histological liver lesions even though it was isolated from a healthy chicken. The results also showed that intrahepatic inoculation of chickens with RNA transcripts of avian HEV infectious clone may serve as an alternative for live virus in animal pathogenicity studies.

Complete genomic sequence of bacteriophage felix o1.

Bacteriophage O1 is a Myoviridae A1 group member used historically for identifying Salmonella. Sequencing revealed a single, linear, 86,155-base-pair genome with 39% average G+C content, 131 open reading frames, and 22 tRNAs. Closest protein homologs occur in Erwinia amylovora phage φEa21-4 and Escherichia coli phage wV8. Proteomic analysis indentified structural proteins: Gp23, Gp36 (major tail protein), Gp49, Gp53, Gp54, Gp55, Gp57, Gp58 (major capsid protein), Gp59, Gp63, Gp64, Gp67, Gp68, Gp69, Gp73, Gp74 and Gp77 (tail fiber). Based on phage-host codon differences, 7 tRNAs could affect translation rate during infection. Introns, holin-lysin cassettes, bacterial toxin homologs and host RNA polymerase-modifying genes were absent.

Comparison of 12 turkey hemorrhagic enteritis virus isolates allows prediction of genetic factors affecting virulence.

Turkey hemorrhagic enteritis virus (THEV) is a member of the genus Siadenovirus and causes disease in turkey poults characterized by splenomegaly, bloody diarrhoea and death. The mechanism responsible for intestinal lesion formation and mortality is not known, although there is strong evidence that it is immune-mediated. All strains of THEV are serologically indistinguishable, although there are naturally occurring avirulent strains of THEV that replicate efficiently in turkeys without the intestinal haemorrhage or mortality associated with more virulent strains. The purpose of this study was to determine which viral genes are involved in virulence. The full-length genome of an avirulent vaccine strain was sequenced and compared with the genome of a virulent field isolate from Israel that was sequenced in 1998. Comparison of the two 26.3 kb genomes revealed 49 nucleotide differences resulting in 14 putative amino acid changes within viral proteins. Sequencing of the regions surrounding the 14 missense mutations revealed variations in ORF1, E3 and the fiber (fib) knob domain in five additional strains with varying degrees of virulence. Complete sequences of these genes were determined in a total of 11 different strains of THEV. All strains had at least one missense mutation in ORF1, and all but two of the strains had one missense mutation in E3. At least one missense mutation was found in the fiber knob domain in six out of seven virulent strains. Sequence variation of ORF1, E3 and fib in strains of THEV with different phenotypes strongly indicates that these genes are the key factors affecting virulence.

Characteristics of diagnostic tests used in the 2002 low-pathogenicity avian influenza H7N2 outbreak in Virginia.

An outbreak of low-pathogenicity avian influenza (LPAI) H7N2 occurred in 2002 in the Shenandoah Valley, a high-density poultry production region in Virginia. Infected flocks were identified through a combination of observation of clinical signs and laboratory diagnostic tests designed to detect avian influenza (AI) antibodies, virus, or H7-specific RNA. In this report, fitness for purpose of 3 virus/RNA detection assays used during the outbreak was examined: 1) antigen capture enzyme immunoassay (AC-EIA), 2) real-time reverse transcription polymerase chain reaction (RRT-PCR), and 3) virus isolation (VI). Results from testing 762 turkey and 2,216 chicken tracheal swab pooled specimens were analyzed to determine diagnostic sensitivities and specificities of these tests under field conditions using Bayesian techniques for validation of diagnostic tests in the absence of a "gold standard." Diagnostic sensitivities (with 95% probability intervals) in turkeys of AC-EIA and RRT-PCR, in reference to VI, were 65.9 (50.6; 81.3)% and 85.1 (71.9; 95.7)% and of VI 92.9 (78.0; 98.8)% in reference to AC-EIA or 88.7 (76.0; 97.2)% in reference to RRT-PCR; in chickens, diagnostic sensitivities were 75.1 (45.6; 94.2)%, 86.3 (65.9; 97.1)%, and 86.2 (65.8; 97.1)% or 86.3 (66.4; 97.2)%, respectively. Specificities were 99.1 (97.9; 99.8)%, 98.9 (98.0; 99.5)%, and 98.6 (97.4; 99.4)% or 98.8 (97.8; 99.5)% in turkeys and between 99.25% and 99.27% with probability intervals of approximately +/-0.4% for all tests in chickens. Simultaneous use of AC-EIA and RRT-PCR contributed significantly to the rapid control of the outbreak, but the AI RRT-PCR assay with >85% sensitivity and approximately 99% specificity, combined with relatively low cost and fast turnaround, could be used as the sole diagnostic test in outbreaks of LPAI.

Identification of new morphological and life-cycle stages of Cochlosoma anatis and experimental transmission using pseudocyst.

Cochlosoma anatis is a flagellated intestinal parasite that infects a variety of avian species. C. anatis infections have been associated with decreased weight gain and increased morbidity and mortality. Conditions favoring the growth of this organism in birds are current pathogenic intestinal infections and/or young age. There is little data describing the life cycle of this parasite. In this study, electron microscopy images are presented that document longitudinal binary fission of the trophozoite stage and outline the events of pseudocyst formation, which includes a rounding stage. Evidence provided here indicates that the pseudocyst stage may be a mechanism for transmission of this organism. The observations reported here provide additional evidence of homology between Cochlosoma and members of the trichomonad order.

Molecular analysis of the relatedness of five domesticated turkey strains.

Our knowledge of the genetic relatedness among the eight existing domesticated turkey strains is limited. To begin to address this paucity, genetic relatedness among five turkey strains (Blue Slate, Bourbon Red, Narragansett, Royal Palm, and Spanish Black) was investigated using three molecular marker systems: randomly amplified polymorphic DNA (RAPD), microsatellite, and SNPs derived from a sequence tagged site and a cloned RAPD fragment. The RAPD analyses were based on five primers that revealed a total of 14 informative DNA fragments in all five populations. The microsatellite analyses involved two informative alleles from three primer-pairs. A total of nine SNPs were detected, one of which appeared to be strain specific. This SNP formed the basis of a PCR-RFLP genotyping procedure developed to distinguish one of the strains from the other four. Evidence from these analyses including the SNP-based RFLP-PCR suggests that Royal Palm is distinct from the other four strains, though more closely related to Narragansett. These data provide, for the first time, molecular evidence of the potential relationships among noncommercial domesticated turkey strains.

Evaluation of risk factors for the spread of low pathogenicity H7N2 avian influenza virus among commercial poultry farms.

OBJECTIVE: To identify risk factors associated with the spread of low pathogenicity H7N2 avian influenza (AI) virus among commercial poultry farms in western Virginia during an outbreak in 2002. DESIGN: Case-control study. PROCEDURE: Questionnaires were used to collect information about farm characteristics, biosecurity measures, and husbandry practices on 151 infected premises (128 turkey and 23 chicken farms) and 199 noninfected premises (167 turkey and 32 chicken farms). RESULTS: The most significant risk factor for AI infection was disposal of dead birds by rendering (odds ratio [OR], 73). In addition, age > or = 10 weeks (OR for birds aged 10 to 19 weeks, 4.9; OR for birds aged > or = 20 weeks, 4.3) was a significant risk factor regardless of poultry species involved. Other significant risk factors included use of nonfamily caretakers and the presence of mammalian wildlife on the farm. Factors that were not significantly associated with infection included use of various routine biosecurity measures, food and litter sources, types of domestic animals on the premises, and presence of wild birds on the premises. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that an important factor contributing to rapid early spread of AI virus infection among commercial poultry farms during this outbreak was disposal of dead birds via rendering off-farm. Because of the highly infectious nature of AI virus and the devastating economic impact of outbreaks, poultry farmers should consider carcass disposal techniques that do not require off-farm movement, such as burial, composting, or incineration.

Suppression of Salmonella growth by wild-type and large-plaque variants of bacteriophage Felix O1 in liquid culture and on chicken frankfurters.

The bacteriophage Felix O1, a member of Myoviridae, is specific for, and possesses a broad host range within, the genus Salmonella. This work explores a Felix O1 phage-based intervention for Salmonella enterica serotype Typhimurium DT104 that is potentially applicable at several stages of animal production and processing. A variant of Felix O1 was obtained that produces a larger, clearer plaque phenotype (LP) on Salmonella Typhi than wild-type Felix O1 (WT) does, not unlike r mutants of phage T4. LP exhibited slightly more extensive overall suppression of Salmonella Typhi in brain heart infusion (BHI) broth, as ascertained on the basis of culture turbidity (optical density at 600 nm). Both phage variants suppressed log phase BHI broth cultures containing 8.2 x 10(6) CFU of Salmonella Typhimurium DT104 per ml. A PFU/CFU ratio of 1.0 was effective for WT and LP, whereas increasing the PFU/CFU ratio to 5.0 did not increase suppression. Untreated Salmonella-contaminated frankfurters were compared with treated samples (PFU/CFU ratio, 1.9 x 10(4)) to test WT and LP for their ability to suppress Salmonella growth on chicken frankfurters contaminated with 300 CFU of Salmonella Typhimurium DT104. Suppression levels of 1.8 and 2.1 log units were achieved with WT and LP, respectively (P = 0.0001), but no difference was found between the performances of the two variants (P = 0.5088).