Therapeutic Efficacy of Phage PIZ SAE-01E2 against Abortion Caused by Salmonella enterica Serovar Abortusequi in Mice.

Salmonella enterica subsp. enterica serovar Abortusequi is a frequently reported pathogen causing abortion in mares. In this study, the preventive and therapeutic effects of phage PIZ SAE-01E2 against S Abortusequi in a mouse model of abortion were investigated. Phage PIZ SAE-01E2 was stable at different temperatures (4 to 70°C) and pH values (pH 4 to 10) and could lyse the majority of the Salmonella serogroup O:4 and O:9 strains tested (25/28). There was no lysogeny-related, toxin, or antibiotic resistance-related gene in the genome of PIZ SAE-01E2. All of these characteristics indicate that PIZ SAE-01E2 has the potential for use in phage therapy. In in vivo experiments, 2 × 103 CFU/mouse of S Abortusequi ATCC 9842 was sufficient to lead to murine abortion (gestational day 14.5) within 48 h. A single intraperitoneal inoculation of PIZ SAE-01E2 (108 PFU/mouse, multiplicity of infection = 105) 1 h before or after S Abortusequi challenge provided effective protection to all pregnant mice (10/10). After 24 h of treatment with phage PIZ SAE-01E2, the bacterial loads in both the placenta and the uterus of the infected mice were significantly decreased (<102 CFU/g) compared to those in the placenta and the uterus of the mice in the control group (>106 CFU/g). In addition, the levels of inflammatory cytokines in the placenta and blood of the mice in the phage administration groups were significantly reduced (P < 0.05) compared to those in the placenta and blood of the mice in the control group. Altogether, these findings indicate that PIZ SAE-01E2 shows the potential to block abortions induced by S Abortusequi in vivoIMPORTANCES Abortusequi is an important pathogen that can induce abortions in mares. Although S Abortusequi has been well controlled in Europe and the United States due to strict breeding and health policies, it is still widespread in African and Asian countries and has proven difficult to control. In China, abortions caused by S Abortusequi have also been reported in donkeys. So far, there is no commercial vaccine. Thus, exploiting alternative efficient and safe strategies to control S Abortusequi infection is essential. In this study, a new lytic phage, PIZ SAE-01E2, infecting S Abortusequi was isolated, and the characteristics of PIZ SAE-01E2 indicated that it has the potential for use in phage therapy. A single intraperitoneal inoculation of PIZ SAE-01E2 before or after S Abortusequi challenge provided effective protection to all pregnant mice. Thus, PIZ SAE-01E2 showed the potential to block abortions induced by S Abortusequi in vivo.

The diagnosis and successful replication of a clinical case of Duck Spleen Necrosis Disease: An experimental co-infection of an emerging unique reovirus and Salmonella indiana reveals the roles of each of the pathogens.

Duck spleen necrosis disease (DSND) is an emerging infectious disease that causes significant economic loss in the duck industry. In 2018, a duck reovirus (named DRV/GX-Y7) and Salmonella indiana were both isolated from the spleens and livers of diseased ducks with DSND in China. The DRV/GX-Y7 strain could propagate in the Vero, LMH, DF-1 and DEF cells with obvious cytopathic effects. The genome of DRV/GX-Y7 was 23,418 bp in length, contained 10 dsRNA segments, ranging from 3959 nt (L1) to 1191 nt (S4). The phylogenetic analysis showed that the DRV/GX-Y7 strain was in the same branch with the new waterfowl-origin reovirus cluster, but was obviously far distant from the clusters of other previous waterfowl-origin reoviruses Muscovy duck reovirus (MDRV) and goose-origin reovirus (GRV), broiler/layer-origin reovirus (ARV) and turkey-origin reovirus (TRV). The RDP and SimPlot program analysis revealed that there were two potential genetic reassortment events in the M2 and S1 segments of the genome. In order to have a clear insight into the pathogenic mechanism of DRV/GX-Y7 and S. Indiana in clinical DSND, an infection experiment was further conducted by challenging commercial ducklings with the two isolates individually and with both. The results showed that DRV/GX-Y7 produced severe hemorrhagic and/or necrotic lesions in the immune organs (thymus, spleen, and bursae) of experimentally infected ducklings. And, that the co-infection of DRV/GX-Y7 and S. Indiana could greatly enhance the pathogenesis by increasing the morbidity and mortality in ducklings whose clinical symptoms and lesions were similar to the natural clinical DSND cases. In summary, the results suggested that the pathogen causing duck spleen necrosis was an emerging unique genetic reassortment strain of duck Orthoreovirus that was significantly different from any previously reported waterfowl-derived Orthoreovirus and the co-infection with the Salmonella isolate could increase the severity of the disease.

Small non-coding RNA STnc640 regulates expression of fimA fimbrial gene and virulence of Salmonella enterica serovar Enteritidis.

BACKGROUND: Small non-coding RNAs (sRNAs) regulate bacterial gene expression at the post-transcriptional level. STnc640 is a type of sRNA that was identified in Salmonella Typhimurium. RESULTS: In this study, STnc640 in Salmonella Enteritidis was confirmed to be an Hfq-dependent sRNA. TargetRNA software analysis showed that fimbrial genes fimA and bcfA were likely to be the target genes of STnc640. To investigate the target mRNAs and function of STnc640 in pathogenicity, we constructed the deletion mutant strain 50336△stnc640 and the complemented strain 50336△stnc640/pstnc640 in Salmonella Enteritidis 50336. The RT-qPCR results showed that the mRNA level of fimA was decreased, while bcfA was unchanged in 50336△stnc640 compared with that in the wild type (WT) strain. The adhesion ability of 50336△stnc640 to Caco-2 cells was increased compared to the 50336 WT strain. The virulence of 50336△stnc640 was enhanced in a one-day-old chicken model of S. Enteritidis disease as determined by quantifying the 50% lethal dose (LD50) of the bacterial strains. CONCLUSIONS: The results demonstrate that STnc640 contributes to the virulence of Salmonella Enteritidis.

Naturally Avian Influenza Virus-Infected Wild Birds Are More Likely to Test Positive for Mycobacterium spp. and Salmonella spp.

Wild birds often harbor infectious microorganisms. Some of these infectious microorganisms may present a risk to domestic animals and humans through spillover events. Detections of certain microorganisms have been shown to increase host susceptibility to infections by other microorganisms, leading to coinfections and altered host-to-host transmission patterns. However, little is known about the frequency of coinfections and its impact on wild bird populations. In order to verify whether avian influenza virus (AIV) natural infection in wild waterbirds was related to the excretion of other microorganisms, 73 AIV-positive samples (feces and cloacal swabs) were coupled with 73 AIV-negative samples of the same sampling characteristics and tested by real-time PCR specific for the following microorganisms: West Nile virus, avian avulavirus 1, Salmonella spp., Yersinia enterocolitica, Yersinia pseudotuberculosis, Mycobacterium avium subspecies, Mycobacterium tuberculosis complex, and Mycobacterium spp. Concurrent detections were found in 47.9% (35/73) of the AIV-positive samples and in 23.3% (17/73) of the AIV-negative samples (P = 0.003). Mycobacterium spp. and Salmonella spp. were found to be significantly more prevalent among the AIV-positive samples than among the AIV-negative samples (42.9% vs. 22.8%; P = 0.024 and 15.2% vs. 0.0%; P = 0.0015, respectively). Prevalence of concurrent detections differed significantly among sampling years (P = 0.001), host families (P = 0.002), host species (P = 0.003), AIV subtypes (P = 0.003), and type of sample (P = 0.009). Multiple concurrent detections (more than one of the tested microorganisms excluding AIV) were found in 9.6% (7/73) of all the AIV-positive samples, accounting for 20% (7/35) of the concurrent detection cases. In contrast, in AIV-negative samples we never detected more than one of the selected microorganisms. These results show that AIV detection was associated with the detection of the monitored microorganisms. Further studies of a larger field sample set or under experimental conditions are necessary to infer causality in these trends.

Las aves silvestres frecuentemente albergan microorganismos infecciosos. Algunos suponen un riesgo por su posible transmisión a animales domésticos o representar un problema de salud pública si son zoonóticos. Se ha relacionado la detección de algunos microorganismos microbianos con una mayor susceptibilidad del hospedador a la infección por otros, llevando a una coinfección y a una alteración de los patrones de transmisión entre hospedadores. Sin embargo, poco se sabe sobre la frecuencia y el impacto de estas coinfecciones en la epidemiologia de las enfermedades en las aves silvestres. Con el ánimo de determinar si una infección natural con el virus de la influenza aviar (VIA) en aves acuáticas se relaciona con la excreción de otros microorganismos, se seleccionaron 73 muestras positivas a VIA y un número igual de negativas de similares características y se sometieron a análisis por PRC a tiempo real para la detección de los siguientes agentes: virus del Nilo occidental, avulavirus aviar de tipo 1, Salmonella spp., Yersinia enterocolitica, Yersinia pseudotuberculosis, subspecies de Mycobacterium avium, complejo Mycobacterium tuberculosis y Mycobacterium spp. Se detectaron otros agentes concurrentes en el 48% (35/73) de las muestras positivas a VIA frente al 23.3% (17/73) en las negativas (p=0.003). La prevalencia de Mycobacterium spp. y Salmonella spp. fue significativamente mayor entre las muestras positivas a VIA que entre las negativas (42.9% vs. 22.8%; p=0.024 y 15.2% vs. 0.0%; p=0.0015 respectivamente). La prevalencia de otros agentes difirió significativamente entre el año de recogida, la familia (p=0.002), la especie (p=0.003), los subtipos de VIA (p=0.003) y el tipo de muestra (p=0.009). Se detectaron múltiples microorganismos en el 9.6% (7/73) de las muestras positivas a VIA, lo que se correspondió con un 20% (7/35) de las detecciones concurrentes. Sin embargo en las muestras negativas a VIA no detectamos más de uno de los microorganismos estudiados. Estos resultados confirman que la detección de los agentes microbianos monitorizados se vio incrementada en presencia del VIA. Consideramos necesario la realización de estudios con un mayor número de muestras o en condiciones experimentales para inferir causalidad sobre estas tendencias.

Outbreaks of antimicrobial resistant Salmonella Choleraesuis in wild boars piglets from central-western Spain.

Salmonella enterica serovar Choleraesuis is the aetiological agent of swine paratyphoid being a highly invasive zoonotic pathogen. Wild boar natural populations are experiencing a demographical expansion as well as some farms are breeding this species to release for hunting with management sometimes identical to that of domestic pigs, including supplementation, grouping, and antibiotic treatments. This situation increases the chance of contact between wild boars and livestock, and potentially induces stress, with different sanitary consequences. The present work aims to describe the clinical features of recent outbreaks caused by S. Choleraesuis in wild boar from central-western Spain, as well as the antimicrobial resistance and phylogenetic relationships of isolates involved. 28 strains of S. Choleraesuis were isolated from 28 different wild boars belonging to 10 different game states located in central western Spain and submitted to the Clinical Veterinary Hospital (CVH) of the University of Extremadura. Samples were taken from different organs and cultured according to the ISO 6579:2002 procedure. Suspicious colonies were identified by PCR and antimicrobial resistance was evaluated by disc diffusion susceptibility test and the presence of the main resistance genes as well as 18 plasmid replicons frequently found among the Enterobacteriaceae was verified by PCR. Pulsed field gel electrophoresis was applied to determine the genetic relationship between isolates. The outbreaks under study were characterized by high mortality (35%-84%) and a septicaemic presentation. S. Choleraesuis was isolated from all the wild boars analysed, and 26 of the 28 isolates presented resistance to at least one antibiotic. The predominant resistances found were against sulphonamide, streptomycin, tetracycline, and doxicicline and sul1, strA-strB, and tetA were the most prevalent resistance genes among isolates. 10 strains carried FIIA, FIB+H/1 or FIIA+H/1 plasmids. PFGE classified the isolates into four different profiles, grouped into two clusters. This results show that prevention against S. Choleraesuis must be considered in the sanitary programs of the wild boar breeders.

[Detection of Salmonella Choleraesuis var. Kunzendorf in a fattening pig with septicaemic salmonellosis. A case report]. / Nachweis von Salmonella Choleraesuis var. Kunzendorf bei einem Mastschwein mit septikämischer Salmonellose. Ein Fallbericht.

The present case reports the detection of Salmonella (S.) Choleraesuis var. Kunzendorf and porcine circovirus type 2 in an organic fattening pig suffering from septicaemic salmonellosis and porcine dermatitis and nephropathy syndrome. Six weeks after pigs had been housed in an organic fattening farm, an increase in mortality, diarrhea and coughing was observed. In recent years, S. choleraesuis var. Kunzendorf has been frequently detected in wild boars in Germany, whereas the same serovar did not play an important role in the Western European domestic pig population. A direct transmission of this serovar from wild boars to domestic pigs in this case could not be proven. However, because wild boars are important reservoirs for the spread of epizootic diseases to the domestic pig population, this case emphasises the importance of taking epidemiological relationships under consideration and to comply with biosecurity measures according to German law (Schweinehaltungshygieneverordnung).

Innate and adaptive immune mechanisms are effectively induced in ileal Peyer's patches of Salmonella typhimurium infected pigs.

In this report we employed laser-capture microdissection (LCM) coupled to qPCR technology and bioinformatic analysis to characterize, for the first time, the response of Peyer's patches (PP) from orally infected animals to Salmonella typhimurium, in a model of non-typhoidal salmonellosis. Pathogen was highly found in the cytoplasm of phagocytes in PP and differential gene expression analysis indicated an up-regulation of proinflammatory molecules, establishment of a Th1 driven response and triggering of DC and T-cell activity. Furthermore, predictions by bioinformatic analysis pointed to an activation of processes regarding stimulation and maturation of DC, influx of leukocytes in tissue and T lymphocytes priming and differentiation. In short, the approach used in this study proved to be a promising strategy to explore infectious processes. Indeed, it revealed an effective induction of innate and adaptive immune mechanisms in swine PP which appear to be distinct from those observed in mesenteric lymph nodes and closely related to response of gut mucosa.

Porcine circovirus type 2 (PCV2) vaccination reduces PCV2 in a PCV2 and Salmonella enterica serovar Choleraesuis coinfection model.

We previously reported that prior porcine circovirus type 2 (PCV2) infection potentiates the severity of clinical signs, lung lesions, and fecal shedding and tissue dissemination of Salmonella enterica serovar Choleraesuis in infected pigs. Here, we evaluated whether PCV2 vaccination is effective in reducing fecal shedding and tissue dissemination of S. Choleraesuis and improving clinical signs associated with PCV2 and S. Choleraesuis infection in 15 Cesarean-derived, colostrum-deprived pigs randomly assigned to 3 groups (n=5/group). The vaccinated and co-infected (VAC-COINF) group received 2 ml of a commercial PCV2 vaccine at age 3 weeks. The VAC-COINF and co-infected (COINF) groups were inoculated intranasally with PCV2 and S. Choleraesuis at 5 and 7 weeks of age, respectively. The CONTROL group pigs received a similar volume of PBS for sham-vaccination and sham-inoculation. PCV2 vaccination clearly reduced PCV2 DNA load in the serum and postmortem tissue samples and decreased PCV2 antigen levels in tissue samples of the VAC-COINF group. After S. Choleraesuis infection, the incidence of several clinical signs increased in the VAC-COINF group compared to that in the COINF group. The microscopic lung lesions and weight gain, fecal shedding and tissue dissemination of S. Choleraesuis except in the spleen were not significantly different in the VAC-COINF and COINF groups. Thus, PCV2 vaccination reduced PCV2 in the S. Choleraesuis and PCV2 coinfection model and the effects on S. Choleraesuis were minimal.

NLRC5 deficiency does not influence cytokine induction by virus and bacteria infections.

Nucleotide-binding domain and leucine rich repeat containing gene family receptors (NLRs) are cytosolic proteins that respond to a variety of pathogen and host components to induce inflammatory cytokines. NLRC5 is a recently identified member of the NLR family that has been implicated in positive and negative regulation of antiviral innate immune responses. To clarify whether NLRC5 controls antiviral innate immunity in vivo, we generated NLRC5-deficient mice. Macrophages and dendritic cells derived from NLRC5-deficient mice induced relatively normal levels of IFN-ß, IL-6, and TNF-α after treatment with RNA viruses, DNA viruses, and bacteria. The serum cytokine levels after polyinosinic-polycytidylic acid infection were also comparable between control and NLRC5-deficient mice. NLRC5 overexpression promoted IL-1ß production via caspase-1, suggesting that NLRC5 constitutes an inflammasome. However, there was no reduction of IL-1ß in NLRC5-deficient cells in response to known inflammasome activators, suggesting that NLRC5 controls IL-1ß production through an unidentified pathway. These findings indicate that NLRC5 is dispensable for cytokine induction in virus and bacterial infections under physiologic conditions.

Evaluation of phage treatment as a strategy to reduce Salmonella populations in growing swine.

Salmonella is a foodborne pathogenic bacterium that causes human illnesses and morbidity and mortality in swine. Bacteriophages are viruses that prey on bacteria and are naturally found in many microbial environments, including the gut of food animals, and have been suggested as a potential intervention strategy to reduce Salmonella levels in the live animal. The present study was designed to determine if anti-Salmonella phages isolated from the feces of commercial finishing swine could reduce gastrointestinal populations of the foodborne pathogen Salmonella Typhimurium in artificially inoculated swine. Weaned pigs (n = 48) were randomly assigned to two treatment groups (control or phage-treated). Each pig was inoculated with Salmonella Typhimurium (2 × 10(10) colony forming units/pig) via oral gavage at 0 h and fecal samples were collected every 24 h. Swine were inoculated with a phage cocktail via oral gavage (3 × 10(9) plaque forming units) at 24 and 48 h. Pigs were humanely killed at 96 h, and cecal and rectal intestinal contents were collected for quantitative and qualitative analysis. Fecal Salmonella populations in phage-treated pigs were lower (p < 0.09) than controls after 48 h. Phage treatment reduced intestinal populations of inoculated Salmonella Typhimurium in pigs compared to controls at necropsy. Cecal populations were reduced (p = 0.07) by phage treatment >1.4 log(10) colony forming units/g digesta, and rectal populations were numerically reduced. The number of pigs that contained inoculated Salmonella Typhimurium was reduced by phage treatment, but a significant (p < 0.05) reduction was only observed in the rectum. We conclude that phages can be a viable tool to reduce Salmonella in swine. Further research needs to be performed to determine the most efficacious dosing regimens and the most effective combinations of phages targeting the diverse Salmonella population found in swine before they can enter the food supply.

Virulence plasmid interchange between strains ATCC 14028, LT2, and SL1344 of Salmonella enterica serovar Typhimurium.

Strains ATCC 14028 and SL1344 of Salmonella enterica serovar Typhimurium are more virulent than LT2 in the BALB/c mouse model. Virulence plasmid swapping between strains ATCC 14208, LT2, and SL1344 does not alter their competitive indexes during mouse infection, indicating that the three plasmids are functionally equivalent, and that their contribution to virulence is independent from the host background. Strains ATCC 14028 and LT2 are more efficient than SL1344 as conjugal donors of the virulence plasmid. Virulence plasmid swapping indicates that reduced ability of conjugal transfer is a property of the SL1344 plasmid, not of the host strain. An A→V amino acid substitution in the TraG protein appears to be the major cause that reduces conjugal transfer in the virulence plasmid of SL1344. Additional sequence differences in the tra operon are found between the SL1344 plasmid and the ATCC 14028 and LT2 plasmids. Divergence in the tra operon may reflect the occurrence of genetic drift either after laboratory domestication or in the environment. The latter might provide evidence that possession of conjugal transfer functions is a neutral trait in Salmonella populations, a view consistent with the abundance of Salmonella isolates whose virulence plasmids are non-conjugative.

Enteric pathology and Salmonella-induced cell death in healthy and SIV-infected rhesus macaques.

The goal of this study was to morphologically characterize a ligated ileal loop model of Salmonella enterica serotype Typhimurium infection in rhesus macaques (Macaca mulatta) and to verify the occurrence of Salmonella-induced cell death in vivo. Eight adult healthy male rhesus macaques were used for ligated ileal loop surgery. Four macaques had been intravenously inoculated with simian immunodeficiency virus (SIV) mac251. Ileal ligated loops were inoculated with wild-type (WT) S. Typhimurium strain IR715 (ATCC14028 nal (r)), an isogenic noninvasive mutant strain (ATCC14028 nal (r) ΔsipAΔsopABDE2), or sterile Luria Bertani broth. Loops were surgically removed at 2, 5, and 8 hours post-inoculation (hpi). Intestinal samples were processed for histopathology, immunohistochemistry for detecting Salmonella, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and transmission electron microscopy. Combined histopathology scores were similar between SIV-infected and control macaques. As expected, the invasion-deficient mutant was less pathogenic than WT S. Typhimurium. Neutrophil infiltrate in the intestinal mucosa correlated with bacterial loads (r = 0.7148; P < .0001) and fluid accumulation (r = 0.6019; P < .0001) in the lumen of the intestinal loops. Immunolabeled WT S. Typhimurium was observed in the epithelium and lamina propria at the tip of the villi at 2 hpi, progressing toward deeper lamina propria at 5-8 hpi. Most TUNEL-positive cells localized to the lamina propria, and some had morphological features of macrophages. Ultrastructurally, bacteria were observed intracellularly in the lamina propria as well as within apoptotic bodies. This study provides morphological evidence of Salmonella-induced cell death in vivo in a relevant nonhuman primate model.

Prevalence of Salmonella infecting bacteriophages associated with Ontario pig farms and the holding area of a high capacity pork processing facility.

BACKGROUND: There is interest in applying bacteriophages to control Salmonella in pig production and pork processing. The following reports on the prevalence of Salmonella infecting bacteriophages within Ontario pig farms and associated with the holding area of a pork slaughterhouse. RESULTS: Salmonella infecting bacteriophages were present in 30 and 28 of the effluent manure samples collected from 36 farms using S. Typhimurium DT104 or S. Heidelberg as host cell respectively. Bacteriophages were recovered in 95-100% of the 48 samples taken from holding pens within a high capacity slaughterhouse over a 12 month period. Bacteriophages isolated from farms exhibited similar host ranges which differed to that of slaughterhouse isolates. Salmonella (n = 21) from the slaughterhouse were susceptible to the endogenous bacteriophages. Despite being susceptible to the resident phages, the Salmonella populations were found to be genetically stable with the same genotypes being recovered over successive visits. Salmonella isolated from the farms were frequently resistant to the endogenous phages. CONCLUSIONS: Bacteriophages are prevalent in the pig slaughterhouse environment although they do not have a significant impact on the genetic structure of Salmonella populations. However, there was evidence that the Salmonella population structure on farms is influenced by the presence of infecting phages.

Salmonella Enteritidis bacteriophage candidates for phage therapy of poultry.

AIMS: Salmonella is a worldwide foodborne pathogen causing acute enteric infections in humans. In the recent years, the use of bacteriophages has been suggested as a possible tool to combat this zoonotic pathogen in poultry farms. This work aims to isolate and perform comparative studies of a group of phages active against a collection of specific Salmonella Enteritidis strains from Portugal and England. Also, suitable phage candidates for therapy of poultry will be selected. METHODS AND RESULTS: The Salm. Enteritidis strains studied were shown to have a significantly high occurrence of defective (cryptic) prophages; however, no live phages were found in the strains. Bacteriophages isolated from different environments lysed all except one of the tested Salm. Enteritidis strains. The bacteriophages studied were divided into different groups according to their genetic homology, RFLP profiles and phenotypic features, and most of them showed no DNA homology with the bacterial hosts. The bacteriophage lytic efficacy proved to be highly dependent on the propagation host strain. CONCLUSIONS: Despite the evidences shown in this work that the Salm. Enteritidis strains used did not produce viable phages, we have confirmed that some phages, when grown on particular hosts, behaved as complexes of phages. This is most likely because of the presence of inactive phage-related genomes (or their parts) in the bacterial strains which are capable of being reactivated or which can recombine with lytic phages. Furthermore, changes of the bacterial hosts used for maintenance of phages must be avoided as these can drastically modify the parameters of the phage preparations, including host range and lytic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: This work shows that the optimal host and growth conditions must be carefully studied and selected for the production of each bacteriophage candidate for animal therapy.

Prevalence of Salmonella spp. antibodies to Toxoplasma gondii, and Newcastle disease virus in feral pigeons (Columba livia) in the city of Jaboticabal, Brazil.

The rock pigeon (Columba livia) may serve as a reservoir for several pathogenic agents that can be transmitted to poultry, wildlife, domesticated pets, and/or humans via excreta, secretions, or dust from feathers. In addition, ingestion of infected pigeons by wild and domestic animals can also transmit these pathogenic agents. The health status of 126 free-living pigeons in an urban area was evaluated by microbiologic culture for Salmonella and serologic testing for the presence of antibodies for Toxoplasma gondii and for Newcastle disease virus (NDV) from 120 and 109 pigeons, respectively. After drawing blood, the birds were euthanized, and fragments of the liver, spleen, lungs, and gonads, and feces were cultured for Salmonella spp. Salmonella spp. was isolated from 10 birds (7.94%), of which 8 were Salmonella typhimurium, one was Salmonella enterica subsp. enterica serotype 4,12 and one was Salmonella enterica subsp. enterica serotype 4,12,i. Six of 109 pigeons (5.50%) were positive for NDV antibodies when using the hemagglutination inhibition test. Toxoplasma gondii antibodies were detected by immunofluorescence in one of 120 sera tested (0.83%). The results indicate that feral rock pigeons were exposed to NDV and T. gondii, although the exposure was low. In addition, these birds had Salmonella spp. and could disseminate this pathogen in the environment.

Multidrug-resistant Salmonella typhimurium, Pacific Northwest, United States.

We compared human and bovine isolates of Salmonella enterica using antimicrobial-drug resistance profiles and pulsed-field gel electrophoresis. From 2000 through 2006, we observed an increase in a novel multidrug-resistant clone of S. Typhimurium with no recognized phage type. This clone may represent an emerging epidemic strain in the Pacific Northwest.

The newly isolated lytic bacteriophages st104a and st104b are highly virulent against Salmonella enterica.

AIMS: To screen Irish faecal samples from a variety of sources with a view to isolating novel anti-Salmonella phages and to subsequently evaluate their lytic capability. METHODS AND RESULTS: Two novel anti-Salmonella phages st104a and st104b were isolated from a screening programme based on their lytic capability. The phages produced significantly larger plaques (2 mm) on the chosen indicator Salmonella enterica strain, DPC6046, when compared with the well-known control phage, Felix 01 (0.5 mm). Both phages st104a and st104b were found to have a broad host range within the Salm. enterica species. During in vitro trials, both phages (st104a and st104b) reduced Salm. enterica numbers more than 99% within 1 h. In vivo studies, involving the addition of the phage to porcine gastric juice (pH 2.5) demonstrated that phage st104a and phage Felix 01 were capable of surviving (10 and 30% survival respectively) the acidic conditions, unlike st104b, which was undetectable after 2 h exposure. CONCLUSIONS: Two novel lytic anti-Salmonella phages were isolated and characterized. SIGNIFICANCE AND IMPACT OF THE STUDY: With the exception of phage Felix 01, there has been relatively little phage therapy work performed using lytic Salmonella phage. In this study, the lytic phages st104a and st104b were isolated as a result of a faecal screening programme. Subsequently, phage st104a was found to have potential for biocontrol of Salm. enterica numbers if administered orally to pigs given their survival in porcine gastric juice, whereas, phage st104b may have potential in reducing cell numbers if applied by alternative approaches.

Homology between two different Salmonella phages: Salmonella enterica serovar Typhimurium phage P22 and Salmonella enterica serovar Anatum var. 15 + phageepsilon34.

A distinguishing feature of many microorganisms, belonging to the Gram negative group of bacteria, is the presence of the lipopolysaccharide on their cell surface. Salmonella is a prominent member of this group of bacteria. Many Salmonella phages use the LPS as the initial receptor in the infection process and they can distinguish subtle changes in the LPS molecules. The phage protein that is responsible for recognition of these cells is the tail or tailspike protein (TSP). Those TSPs, which use LPS as a receptor, are prokaryotic LPS-binding proteins. As an initial step in using phage TSPs as model systems for a detailed molecular genetic analysis of protein-LPS interactions, a comparison of two phages and their TSPs from two different Salmonella bacterial viruses (phages), Salmonella enterica serovar Typhimurium phage P22 and Salmonella enterica serovar Anatum var. 15 + phage epsilon34, is being carried out. This present study shows significant viral protein homology between many viral structural proteins from these two phages including their TSPs. Significantly this report suggests a general structural motif for part of the TSP of phages and suggests that a more detailed comparative analysis of these TSPs is warranted.

A questionnaire-based, retrospective field study of persistence of Salmonella Enteritidis and Salmonella Typhimurium in Danish broiler houses.

A questionnaire-based, retrospective field study was conducted in 78 Danish broiler houses (analytical units) on 42 farms. In spring 1997, all these broiler houses had been infected with Salmonella Enteritidis, phage type 8, and/or Salmonella Typhimurium, definitive-type 66, by day-old chicks delivered from the same hatchery. Because these two salmonella types had not been detected in the Danish broiler sector before and the broiler houses were infected in a period of low salmonella prevalence, the study could focus on farm-related factors, without being distorted by other salmonella infections. The same person visited all 78 broiler houses, and farm-related factors were listed both from personal observations and by interrogating the person in charge of attending to the broilers. Additional factors (noted at the visits) were recorded in a later telephone interview. Altogether, 187 variables were analyzed.Broiler houses were divided into single-infection houses (which had one or both of the salmonella types in only one crop), and multiple-infection houses (where infection occurred in multiple crops). This was done both generally and separately for each of the salmonella types. Factors associated with house status-but that were beyond the control of the farmer-were the salmonella type and the percentage of salmonella positive samples in the first crop. Factors which could be controlled by the farmer and which were associated with reduced prevalence of multiple-infection houses included antiseptic soap and water for washing hands in the anteroom, hygiene barriers when removing dead broilers, gravel alongside the broiler house, systematic checks of indoor rodent-bait depots, and combined surface and pulse-fogging disinfection.

Deposition of phage type 4 and 13a Salmonella enteritidis strains in the yolk and albumen of eggs laid by experimentally infected hens.

Because egg yolk and albumen differ substantially in their abilities to support bacterial growth, the initial level and location of Salmonella enteritidis deposition are critical for determining whether proposed standards for refrigerating eggs are likely to protect public health by preventing extensive microbial multiplication. In the present study, three groups of laying hens were infected with oral doses of approximately 10(9) cells of different S. enteritidis strains (two were phage type 4 and one was phage type 13a) in two replicate trials. For all three S. enteritidis strains, the incidence of yolk contamination (approximately 2.5% overall) was significantly greater than the incidence of albumen contamination (approximately 0.5% overall). The phage type 13a strain was less often isolated from fecal samples at 2 wk post-inoculation than were the phage type 4 strains, but no significant differences between strains were observed in the incidence of egg contamination. Most freshly laid contaminated eggs contained fewer than 1 S. enteritidis cell/ml of egg yolk or albumen, and no sample contained more than 67 S. enteritidis cells/ml.