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1.
The cross-reactivity of egg yolk antibodies specific to antigens of Salmonella Enteritidis and Salmonella Typhimurium to killed bacterial cells of common Salmonella serovars were tested using an indirect Enzyme Linked Immunosorbent Assay (ELISA). Egg yolk antibodies were produced against purified fimbriae, flagella and lipopolysaccharide (LPS) of S. Enteritidis strain ATCC13076 and flagella, LPS and outer membrane proteins (OMP) of S. Typhimurium strain ATCC13311. For immunological specificity of egg yolk antibodies against killed bacterial cells, we found that the titers of the anti-S. Enteritidis egg yolk antibodies were higher than those of the anti-S. Typhimurium antibodies. In the evaluation of cross-reactivity of these egg yolk antibodies to various Salmonella serovars, we observed that the anti-S. Enteritidis antibodies exhibited more specific affinity than those of the anti-S. Typhimurium antibodies. All S. Enteritidis strains reacted specifically with the anti-S. Enteritidis fimbrial and flagellar egg yolk antibody whereas anti-S. Enteritidis LPS and anti-S. Typhimurium LPS, OMP and flagellar antibodies displayed non–specific reactivity to all Salmonella serovars used in this study. This finding suggests that it may be possible to design a anti-fimbrial egg yolk antibody of S. Enteritidis as a diagnostic tool and a cocktail of OMP and LPS antigens of S. Enteritidis and S. Typhimurium could be used for administering broad spectrum passive immunity to protect against the colonization of pathogenic Salmonella strains in food animals.  相似文献   

2.
Salmonella serovars, one of the leading contributors to foodborne illness and are especially problematic for foods that are not cooked before consumption, such as fresh produce. The shipping containers that are used to transport and store fresh produce may play a role in cross contamination and subsequent illnesses. However, methods for quantitatively attached cells are somewhat variable. The overall goal of this study was to compare conventional plating with molecular methods for quantitating attached representative strains for Salmonella Typhimurium and Heidelberg on reusable plastic containers (RPC) coupons, respectively. We attached Salmonella enterica serovar Typhimurium ATCC 14028 and serovar Heidelberg SL486 (parent and an antibiotic resistant marker strain) to plastic coupons (2.54 cm2) derived from previously used shipping containers by growing for 72 h in tryptic soy broth. The impact of the concentration of sanitizer on log reductions between unsanitized and sanitized coupons was evaluated by exposing attached S. Typhimurium cells to 200 ppm and 200,000 ppm sodium hypochlorite (NaClO). Differences in sanitizer effectiveness between serovars were also evaluated with attached S. Typhimurium compared to attached S. Heidelberg populations after being exposed to 200 ppm peracetic acid (PAA). Treatment with NaClO caused an average of 2.73 ± 0.23 log CFU of S. Typhimurium per coupon removed with treatment at 200 ppm while 3.36 ± 0.54 log CFU were removed at 200,000 ppm. Treatment with PAA caused an average of 2.62 ± 0.15 log CFU removed for S. Typhimurium and 1.41 ± 0.17 log CFU for S. Heidelberg (parent) and 1.61 ± 0.08 log CFU (marker). Lastly, scanning electron microscopy (SEM) was used to visualize cell attachment and coupon surface topography. SEM images showed that remaining attached cell populations were visible even after sanitizer application. Conventional plating and qPCR yielded similar levels of enumerated bacterial populations indicating a high concordance between the two methods. Therefore, qPCR could be used for the rapid quantification of Salmonella attached on RPC.  相似文献   

3.
Environmental Science and Pollution Research - Unsuccessful vaccination against Salmonella due to a large number of serovars, and antibiotic resistance, necessitates the development of novel...  相似文献   

4.
The present study investigated SCFA exposed Salmonella enterica serovar Typhimurium for its virulence characteristics such as mouse lethality adhesion, surface hydrophobicity, phagocytic uptake, intracellular survival within the murine peritoneal macrophages, induction of acid tolerance response (ATR), and the expression of outer membrane proteins (OMPs). Lethal dose (LD50) for mice was found to be more for SCFA exposed cells as compared to the normal cells. In vitro adherence to murine intestinal epithelial cells as well as surface hydrophobicity of SCFA exposed cells were found to be lower than the normal cells. Though the phagocytic uptake of normal cells and SCFA exposed cells was similar, the SCFA exposed cells exhibited increased intracellular survival as compared to the normal cells, which were completely killed after 4 h. Moreover, SCFA exposed cells also survived exposure to extremely low pH (3.0). Outer membrane proteins extracted from SCFA exposed cells revealed an enhanced expression of proteins (porins) at 4% SCFA concentration. Thus, SCFA exposure may contribute to enhanced virulence of Salmonella enterica serovar Typhimurium by increased intracellular survival, induction of acid tolerance response, and the enhanced expression of outer membrane matrix proteins (porins).  相似文献   

5.
The objective of the present study was to investigate the ability of animal feed-grade sodium bisulfate (SBS) and a mixture of sodium bisulfate/tannin to inhibit the growth of Salmonella using an anerobic in vitro mixed cecal culture to mimic the conditions within the chicken cecum. An initial inoculum of Salmonella Typhimurium was introduced to an anerobic dilution solution containing 1/3000 diluted cecal bacteria and solids consisting of ground chicken feed and different percentages of solid SBS or SBS/tannin, and surviving organisms were enumerated. Two different experimental designs were employed. In the “unadapted” treatment, the S. Typhimurium was added at the beginning of the culture incubation along with cecal bacteria and chicken feed/SBS or chicken feed/SBS/tannin. In the “adapted” treatment, S. Typhimurium was added after a 24 hour pre-incubation of the cecal bacteria with the chicken feed/SBS or chicken feed/SBS/tannin. Adding SBS resulted in reduction of pH in the cultures which paralleled with the reduction of S. Typhimurium. The SBS alone was found to be inhibitory to S. Typhimurium in the adapted treatment at all concentrations tested (0.25, 0.5, and 0.75%), and the degree of inhibition was concentration-dependent. Salmonella Typhimurium was completely killed in the adapted culture with 0.5% SBS after 24 and 48 h. The SBS/tannin mixture was less inhibitory than SBS alone at the same concentrations in side-by-side comparisons. Testing at a 0.5% SBS concentration, chicken age had little or no effect on log reduction of S. Typhimurium relative to age-matched control cultures without SBS, but age did affect the absolute number of S. Typhimurium surviving, with the greatest decreases occurring at 2 and 4 weeks of age (approx. 103 S. Typhimurium surviving) compared to 6 weeks of age (approx. 105 Salmonella surviving). Microbiome analysis with an Illumina MiSeq platform was conducted to investigate bacterial compositional changes related to the addition of SBS. The relative abundance of Firmicutes (at the phylum level) was decreased, and genera Lactobacillus and Faecalibacterium were increased when SBS was added to the anaerobic mixed culture containing either fecal or cecal material. The antimicrobial action of feed-grade SBS may represent a potential pre-harvest control measure for Salmonella in poultry production.  相似文献   

6.
The chick embryotoxicity screening test (CHEST) and the Salmonella/microsome bioassay were used to evaluate embryotoxic and mutagenic endpoints from crude coal tar (CT) and its fractionated polycyclic aromatic hydrocarbon (PAH) mixtures (designated as A, B, C, D and E). In the CHEST assay, CT and PAH mixtures were injected into the egg yolk. A dose-dependent increase in embryo mortality was observed for all fractions. The E fraction resulted in 47% embryo mortality at a dose of 0.125 mg/kg and was more toxic than CT. At a dose of 1 mg/kg, 85-100% embryonic deaths occurred in fractions C and D and these two fractions were more potent than fractions A and B. The main visual toxic manifestations were liver lesions, discoloration of the liver, and edema. Both CT and fractionated PAH mixtures were also tested in the Salmonella/microsome plate incorporation assay with Salmonella typhimurium strain TA98 and were evaluated with and without metabolic activation at five dose levels. In the presence of S9, the CT and fractions C, D and E induced a dose-dependent positive response. Results from the Salmonella/microsome assay were in good agreement with findings from the CHEST assay suggesting that these two bioassays in combination may facilitate the rapid detection and ranking of complex PAH mixtures.  相似文献   

7.
The objective of this study was to determine the effectiveness of an iodine based disinfectant (IBD, Iocide, Biomedical Development Corporation, San Antonio, TX) on Salmonella enteritidis and S. typhimurium inoculated on egg shell surfaces under simulated industry egg processing conditions with a commercial egg washer used as the sanitizer delivery system. Re-circulated egg washer water containing 1.40-2.85 g/l total dissolved solids was obtained from a commercial egg processing. Sanitizing treatments consisted of distilled deionized water (DDW), IBD, and chlorine (CL; 200 ppm). All treatments (DDW, IBD and CL) significantly (p < 0.05) decreased Salmonella spp. populations on the shell compared to dry (no spray) egg controls. However, efficacy of egg sanitizers appeared to be dependent on the level of total dissolved solids in the egg wash water.  相似文献   

8.
Virulence expression of Salmonella enterica serovar Typhimurium under iron limited condition was measured by beta-galactosidase (beta-gal) assay using a hilA-lacZY fusion strain and calculated as Miller units. hilA-lacZY beta-galactosidase assays were performed in brain heart infusion (BHI) and minimal media (M9), after iron chelation with 2, 2-dipridyl and iron-supplementation respectively. Before performing virulence assays, concentrations of iron in the media were estimated using ferrozine. Iron content was found to be more in BHI (42.6 microg dL(-1)) as compared to M9 (10.03 microg dL(-1)). beta-gal activity of Salmonella Typhimurium in BHI was generally less than that observed in M9. After exposure to various combinations of iron chelator in BHI, hilA-lacZY activity only increased at the highest concentration of chelator (2001 microM) but decreased in M9 media for all iron concentrations when compared to controls with no iron amendment. These results indicate that iron availability may influence S. Typhimurium hilA expression.  相似文献   

9.
The capabilities of selected strains from genera Lactobacillus and Bifidobacterium to produce extracellular bioactive compounds with antimutagenic properties against benzo[a]pyrene (BaP) and sodium azide (SA) were tested as a function of growth phase. The bacterial supernatants from exponential and stationary phases were characterized with different patterns of antimutagenic activity against the two mutagens. All lactobacilli exhibited either no effect or low antimutagenicity against BaP during exponential growth. Higher antimutagenic activities of lactobacilli supernatants were observed in the stationary phase against SA as well. An exception was Lactobacillus sakei 23K which expressed a relatively low percent of inhibition of mutagenesis (PI = 28.14 +/- 7.41) in the exponential phase and no antimutagenic activity in the stationary phase. Of the bifidobacteria, only Bifidobacterium adoleascentis ATCC 15703 exhibited higher antimutagenecity against BaP in the exponential phase. The same bacterial supernatants however, did not possess any antimutagenicity against SA in either the exponential or stationary phases. B. bifidum ATCC 11863 did not express any significant differences in its activity against either BaP or SA in the exponential or stationary phases. Only B. breve ATCC 15700 expressed a high antimutagenic effect against SA in the stationary phase but exhibited no effect during exponential growth. Overall, bacterial antimutagenic responses were associated with growth phase and type of mutagen.  相似文献   

10.
The capabilities of selected strains from genera Lactobacillus and Bifidobacterium to produce extracellular bioactive compounds with antimutagenic properties against benzo[a]pyrene (BaP) and sodium azide (SA) were tested as a function of growth phase. The bacterial supernatants from exponential and stationary phases were characterized with different patterns of antimutagenic activity against the two mutagens. All lactobacilli exhibited either no effect or low antimutagenicity against BaP during exponential growth. Higher antimutagenic activities of lactobacilli supernatants were observed in the stationary phase against SA as well. An exception was Lactobacillus sakei 23K which expressed a relatively low percent of inhibition of mutagenesis (PI = 28.14 ± 7.41) in the exponential phase and no antimutagenic activity in the stationary phase. Of the bifidobacteria, only Bifidobacterium adoleascentis ATCC 15703 exhibited higher antimutagenecity against BaP in the exponential phase. The same bacterial supernatants however, did not possess any antimutagenicity against SA in either the exponential or stationary phases. B. bifidum ATCC 11863 did not express any significant differences in its activity against either BaP or SA in the exponential or stationary phases. Only B. breve ATCC 15700 expressed a high antimutagenic effect against SA in the stationary phase but exhibited no effect during exponential growth. Overall, bacterial antimutagenic responses were associated with growth phase and type of mutagen.  相似文献   

11.
Laying hens were treated orally with a single dose of aldrin (1,2,3,4,10,10-hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5,8-dimethanonaphthalene, AD) 1 mg kg(-1) bw. Concentrations (microg g(-1)) of AD or its epoxide, dieldrin (1,2,3,4,10,10-hexachloro-6,7-epoxy-1,4,4a,5,6,7,8,8a-octahydroendo-,exo-1,4:5,8-dimetha-nonaphthalene, DD), in the main tissues involved in egg formation (blood, liver, ovary, and oviducts) and egg yolk, collected at 1 day after AD dosing, were determined by normal-phase high-performance liquid chromatography. The limits of determination were 0.07 microg g(-1) for AD and 0.08 microg g(-1) for DD, respectively. In extractable fats from the above tissues and egg yolk, AD was found in the egg yolk; however, no AD was found in tissues involved in egg formation. DD was found in all tissues examined here. The DD level was highest in the liver and was lowest in the blood (P<0.01). These results suggest that the epoxidation of AD to DD occurred rapidly in the hen.  相似文献   

12.
The objective of this study was to screen the genome of Salmonella typhimurium for genes potentially required for survival on egg shell surfaces using a novel transposon footprinting method based on a previously developed Tn5 mutagenesis technique. Since the amplified DNA fragment length should usually be unique for each mutant, the polymerase chain reaction products separated on an agarose gel generate a transposon footprint with each band in the footprint representing the corresponding Tn5 mutant. By comparing the footprints from the pools of S. typhimurium Tn5 mutants before and after inoculation on egg shell surfaces, Tn5 mutants not recovered after selection were rapidly identified for potential isolation and genetic analysis.  相似文献   

13.
Abstract

Virulence expression of Salmonella enterica serovar Typhimurium under iron limited condition was measured by β-galactosidase (β-gal) assay using a hilA-lacZY fusion strain and calculated as Miller units. hilA-lacZY β-galactosidase assays were performed in brain heart infusion (BHI) and minimal media (M9), after iron chelation with 2, 2-dipridyl and iron-supplementation respectively. Before performing virulence assays, concentrations of iron in the media were estimated using ferrozine. Iron content was found to be more in BHI (42.6 µg dL?1) as compared to M9 (10.03 µg dL?1). β-gal activity of Salmonella Typhimurium in BHI was generally less than that observed in M9. After exposure to various combinations of iron chelator in BHI, hilA-lacZY activity only increased at the highest concentration of chelator (200 µM) but decreased in M9 media for all iron concentrations when compared to controls with no iron amendment. These results indicate that iron availability may influence S. Typhimurium hilA expression.  相似文献   

14.
Polychlorinated biphenyls (PCBs) are persistent and hazardous environmental contaminants, which tend to bioaccumulate in the food chain. In the present report the long-term effect of low-level dietary PCB concentrations was studied on performance, egg quality, apparent PCB digestibility, apparent PCB retention and PCB accumulation in laying hens that were fed experimental diets for 41 weeks. The tested dietary concentrations of supplemented PCBs, based on the sum of seven reference congeners, were 0, 1.5 and 6 ng/g. PCB ingestion did not significantly affect performance or egg quality parameters. The PCB concentration in egg yolk reached a nearly constant level after approximately 40 and 70 days of consumption of the diets containing 1.5 and 6 ng PCBs/g, respectively. Apparent faecal PCB digestibility and apparent retention were not influenced by dietary levels of added fat varying between 1.5% and 4.5%, but were significantly higher in hens fed diets containing added PCBs. Moreover, apparent PCB digestibility and retention increased significantly with age. Among the seven individual PCB congeners, no systematically significant differences with regard to apparent faecal digestibility were observed throughout the experiment. Accumulation of PCBs in the fat fraction of egg yolk, abdominal adipose tissue and thigh and breast muscle greatly depended upon PCB intake, but never exceeded the maximally allowed concentration of 200 ng/g. As PCBs 52 and 101 were hardly found in egg yolks and hen tissues, it was concluded that both congeners were greatly metabolised. Comparison of relative contents of individual PCB congeners revealed that PCBs 118, 138 and 153 were preferentially incorporated in yolk and body tissues.  相似文献   

15.
The aim of this study was to test a specific method for the detection of Vitellogenin (Vtg) and Zona Radiata Proteins (Zrp) in plasma from peregrine falcon (Falco peregrinus) as specific biomarkers for the evaluation of the effects of endocrine disruptors. The method was assayed with different peregrine falcon individuals (including mature and immature birds of both sexes) from a Spanish population being studied in terms of their contamination with organochlorine compounds with endocrine disrupting properties. This study shows that mouse anti bird Vtg monoclonal antibody ND3C3 (Biosense) seems to be the most specific antibody in binding plasmatic lipoproteins in peregrine falcon when compared to other anti Vtg antibodies. Rabbit anti salmon Zrp polyclonal antibodies O146 (Biosense) show cross-reactivity with Zrp in the samples studied. These preliminary results confirm the applicability of both of these diagnostic tools assayed (induction of Vtg and Zrp) in detecting exposure to Endocrine Disrupting Chemicals (EDCs) in this species. The increase of Vtg and Zrp detected in male specimens suggest a potential hazard to EDCs in the peregrine falcon which represents a species still affected by organochlorine compounds, and in particular those with estrogenic activity.  相似文献   

16.
A rapid Biacore biosensor immunoassay of 4-nonylphenols was developed. Two types of antibodies were used in the study: polyclonal antibodies with high cross-reactivity towards technical 4-nonylphenol and a monoclonal antibody very specific to 4-n-nonylphenol. 9-(p-Hydroxyphenyl)nonanoic acid was immobilized onto surface of a sensor chip. The best assay sensitivity was achieved using a flow rate of 50 microl min(-1) and injection time of 2 min. For the assay incorporating monoclonal antibodies a limit of detection 2 ng ml(-1) for 4-n-nonylphenol was achieved. With polyclonal antibodies one order lower sensitivity was observed for 4-nonylphenols. High background level of calibration curve for technical 4-nonylphenol was decreased by using IgG fraction of polyclonal antibodies in combination with lower amount of immobilised 9-(p-hydroxyphenyl)nonanoic acid. Sensitivity of the assay was improved by using a chip with a new derivative on a surface-N-aminobutyl [2-(4-hydroxyphenyl)ethylamine] (limit of detection--5 ng ml(-1)). Applicability of the developed assays to ecological monitoring was checked in experiments using shellfish samples. 4-n-Nonylphenol from spiked samples was extracted into hexane followed by clean-up on NH2 SPE columns. Calibration curves generated for cockles, mussels and oyster samples were identical (limit of detection about 10 ng g(-1)) whereas for scallop samples a slight decrease (about 5-10%) of absolute response was observed. In the assay using the monoclonal antibody specific to 4-n-nonylphenol 31 shellfish samples were found to be negative. Results obtained with polyclonal antibodies indicated that two scallop samples contained a quantity of 4-nonylphenols. The developed biosensor assay could be applied for shellfish analysis as a preliminary screening method.  相似文献   

17.
Genotoxicity of the insecticide methyl parathion was investigated in Salmonella typhimurium and Escherichia coli bacterial test systems for the detection of back mutations and DNA-damage. Methyl parathion was mutagenic to S. typhimurium strain TA100 after activation with rat liver microsomal and cytosolic enzymes. In DNA repair tests, methyl parathion was effective in inducing damage to the S. typhimurium strain TA1538 which lack excision repair compared to the strain TA1978 which is proficient in excision repair mechanisms. Normal laboratory light conditions had no effect on the mutagenicity tests, however, exposure of methyl parathion in the petri dish containing the tester strain TA100 and rat liver microsomal and cytosolic enzymes reduced the mutagenic activity and increased the toxic effects of methyl parathion.  相似文献   

18.
The antifungal activity of Artemisia herba alba was found to be associated with two major volatile compounds isolated from the fresh leaves of the plant. Carvone and piperitone were isolated and identified by GC/MS, GC/IR, and NMR spectroscopy. Antifungal activity was measured against Penicillium citrinum (ATCC 10499) and Mucora rouxii (ATCC 24905). The antifungal activity (IC50) of the purified compounds was estimated to be 5 microg/ml, 2 microg/ml against Penicillium citrinum and 7 microg/ml, 1.5 microg/ml against Mucora rouxii carvone and piperitone, respectively.  相似文献   

19.
The presence of Maillard reaction products (MRP) in foods and food components is due to the non-enzymatic reaction between protein and carbohydrate residues triggered by thermal steps during food processing. The objective of this study was to assess the effect of MRPs and increasing lysine concentrations on S. Typhimurium growth and the expression of cadA which may be an indirect determinant of Salmonella virulence response. Variations in lysine concentrations (from 0 to 0.5 mM) did not exert any effect either on the final optical density after 6-hour incubation or the growth rates of S. Typhimurium in media containing MRPs. In contrast to the reduced final absorbancy of the bacterial cultures grown with histidine and arginine MRPs supplementations (0.1%), growth rates, in general, remained unaltered by all MRPs at each lysine concentration when compared to the control (M9 pH 5.8, no MRPs added). The induction levels of cadA in media containing 0.1% MRPs were close to cadA induction in the reference media (M9, pH 5.8 and no MRPs) and did not exceed the corresponding values by more than approximately 30%. Although the observed negligible induction of cadA under these conditions complies with the concept of its potential “anti-virulence” function, additional studies involving various concentrations and more refined MRPs are needed.  相似文献   

20.
Abstract

There is limited research concerning the biofilm-forming capabilities of Salmonella Kentucky, a common poultry isolate. The objective was to quantitate pellicle formation of S. Kentucky versus better-characterized Salmonella strains of Enteritidis and Heidelberg. In separate experiments, Salmonella strains and serovars were tested for their biofilm-forming abilities in different Luria-Bertani (LB) broths (1); pellicle formation in different volumes of LB without salt (2); and the potential priming effects on formation after pellicles were transferred three consecutive times (3). Data were analyzed using One-Way ANOVA with means separated using Tukey’s HSD (P?≤?0.05). In the first experiment, there was no significant effect between strain and serovars (P?>?0.05), but media type affected pellicle formation significantly with LB Miller and LB minus NaCl plus 2% glucose resulting in no pellicle formation (P?<?0.001). When grown in 50?mL, Kentucky 38-0085 produced larger pellicles than Kentucky 38-0055, and Heidelberg strain 38-0127 (P?<?0.0001). Serial transfers of pellicles did not significantly affect pellicle formation (P?>?0.05); however, Kentucky 38-0084, 38-0085 and 38-0086 produced larger pellicles than Kentucky 38-0055 and 38-0056 and Heidelberg 38-0126, 38-0127 and 38-0152. The current study demonstrates the consistent biofilm forming capabilities of Kentucky and may explain why Kentucky is frequently isolated in poultry processing facilities.  相似文献   

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