The Combination of Zinc Compounds and Acidic pH Limits Aerobic Growth of a Salmonella typhimurium Poultry Marker Strain in Rich and Minimal Media

Abstract

The objective of the present study was to examine the combined effects of zinc compounds with different acidic pH levels on the aerobic growth of a S. typhimurium poultry isolate in either rich or minimal media. When overall main effects of pH levels of medium or concentrations of Zn compounds were compared, growth rates of the S. typhimurium poultry isolate were significantly ( p < 0.05) decreased by stepwise increase of pH levels of medium (pH 4, 5, 6, and 7) or concentrations (0.67, 3.35, and 6.03%) of Zn compounds (Zn acetate and Zn sulfate). In general growth rates of S. typhimurium poultry isolate appeared to be more reduced by Zn acetate than by Zn sulfate and more reduced in minimal media compared to rich media.     

Response of selected poultry cecal probiotic bacteria and a primary poultry Salmonella typhimurium isolate grown with or without glucose in liquid batch culture

The objective of this study was to determine whether fermentation by a cecal probiotic co-culture of an Enterococcus sp. and Veillonella sp. would inhibit the in vitro growth of a S. typhimurium poultry isolate. The growth rates of S. typhimurium and Enterococcus were significantly reduced at pH 5. At the two pH levels, there was a significant (p < 0.001) increase at 24 h in colony forming units for each of the bacteria enumerated from the mixed culture compared to the respective pure culture enumerations. S. typhimurium was not inhibited in mixed cultures. The mixed cultures produced more acetate than any of the pure cultures and lactate produced by Enterococcus appeared to be utilized by Veillonella.     

Response of selected poultry cecal probiotic bacteria and a primary poultry salmonella typhimurium isolate grown with or without glucose in liquid batch culture

Abstract

The objective of this study was to determine whether fermentation by a cecal probiotic co‐culture of an Enterococcus sp. and Veillonella sp. would inhibit the in vitro growth of a S typhimurium poultry isolate. The growth rates of S. typhimurium and Enterococcus were significantly reduced at pH 5. At the two pH levels, there was a significant (p < 0.001) increase at 24 h in colony forming units for each of the bacteria enumerated from the mixed culture compared to the respective pure culture enumerations. S. typhimurium was not inhibited in mixed cultures. The mixed cultures produced more acetate than any of the pure cultures and lactate produced by Enterococcus appeared to be utilized by Veillonella.     

Short-chain fatty acids affect cell-association and invasion of HEp-2 cells by Salmonella typhimurium

This study demonstrates that the growth of S. typhimurium in Luria Bertani broth supplemented with acetate, propionate, butyrate, or a mixture of the three SCFA, affected cell-association and the ability to invade cultured HEp-2 cells. Cell-association and invasion was determined after growth for 4 h of growth in the presence of the SCFA at pH 6 and 7. The results suggest that the growth rate of the culture may have affected cell-association and invasion since accompanying the significant decrease in growth rate in the presence of SCFA at pH 6 was a decrease in cell-association and invasion. However, the results also suggest that the individual SCFA may play a role in modulating cell-association and the invasion phenotype and the regulation of cell-association and invasion by the SCFA was dependent on the concentration and the pH of the medium. Although the growth rates were similar for S. typhimurium in the SCFA mixture, butyrate (100 mM) and propionate (50 mM) at pH 6, differences in cell-association and invasion were observed among these cultures. Also, at pH 7, differences were observed among the SCFA treatments even though the growth rates were similar.     

Copper and Zn uptake by radish and pakchoi as affected by application of livestock and poultry manures

Environmental safety of agricultural utilization of livestock and poultry manures from intensive farming is attracting great attention because the manures often contain high concentrations of heavy metals and organic pollutants. Pot experiments, in which a pig manure (PM), a chicken manure (CM) and a commercial organic manure (OM) with different concentrations of Cu and Zn to simulate soil metal accumulation by manure application for different times were utilized in a garden soil at a rate of 2% (W/W), were conducted to study the effect of application of these livestock and poultry manures on growth of radish (Raphanus sativus L.) and pakchoi (Brassica chinensis L.) as well as their Cu and Zn uptake. The results exhibit that the manures except the PM improved the growth of radish and pakchoi. The difference of biomass among the same manure treatments containing different concentrations of Cu and Zn, however, was insignificant. In addition, application of the livestock and poultry manures significantly increased soil pHs and electric conductivities (EC) compared with the control, which is ascribed that these manures had high pH and contained large amounts of inorganic ions. The available soil Zn concentrations in the PM were higher than that in the CM and OM, and the extractable soil Cu concentrations in the three manures were almost the same after radish growth in the garden soil but were different after pakchoi growth. Zinc and Cu concentrations in the radish and pakchoi tissues increased when the soil Zn and Cu concentrations increased by manures application, but were still within a safe value. An except is the treatment PM4 in which the Zn concentration of the above-ground part of radish was 28.7 mg kg-1, exceeding the Chinese Food Hygiene Standard of 20 mg kg-1 based on fresh weight. Good correlation was obtained between the extractable soil Zn (or Cu) concentrations extracted by 1.0 mol l-1 NH4NO3 and the Zn (or Cu) concentrations in radish and pakchoi tissues, which was expected to be effective in forecasting Cu and Zn availability to radish and pakchoi in manure agronomic utilization.     

Recovery of a marker strain of Salmonella typhimurium in litter and aerosols from isolation rooms containing infected chickens

Screening of poultry flocks for foodborne pathogen Salmonella contamination is critical for Salmonella control in preharvest stages of poultry production. In this study, two sampling methods (litter and air filter) were compared for detection of S. typhimurium from experimentally infected chicks some of which had received either a probiotic competitive exclusion culture or transfer of cecal contents from salmonellae-free adult birds. At 4, 9, and 11 days after inoculation, S. typhimurium samples were enumerated by selective plating. For both types of sampling, the control birds yielded the greatest levels of environmental contamination followed by the samples from the probiotic inoculated birds with the birds receiving the cecal transfer culture having the lowest levels of contamination. Although the two sampling methods responded in a similar fashion, detection sensitivity needs to be increased for air filter sampling.     

Effect of soluble maillard reaction products on cadA expression in Salmonella typhimurium

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.     

Sulfur toxicity and media capacity for H2S removal in biofilters packed with a natural or a commercial granular medium

Two types of media, a natural medium (wood chips) and a commercially engineered medium, were evaluated for sulfur inhibition and capacity for removal of hydrogen sulfide (H2S). Sulfate was added artificially (40, 65, and 100 mg of S/g of medium) to test its effect on removal efficiency and the media. A humidified gas stream of 50 ppm by volume H2S was passed through the media-packed columns, and effluent readings for H2S at the outlet were measured continuously. The overall H2S baseline removal efficiencies of the column packed with natural medium remained >95% over a 2-day period even with the accumulated sulfur species. Added sulfate at a concentration high enough to saturate the biofilter moisture phase did not appear to affect the H2S removal process efficiency. The results of additional experiments with a commercial granular medium also demonstrated that the accumulation of amounts of sulfate sufficient enough to saturate the moisture phase of the medium did not have a significant effect on H2S removal. When the pH of the biofilter medium was lowered to 4, H2S removal efficiency did drop to 36%. This work suggests that sulfate mass transfer through the moisture phase to the biofilm phase does not appear to inhibit H2S removal rates in biofilters. Thus, performance degradation for odor-removing biofilters or H2S breakthrough in field applications is probably caused by other consequences of high H2S loading, such as sulfur precipitation.     

Composition of aqueous extracts of broiler litter treated with aluminum sulfate, ferrous sulfate, ferric chloride and gypsum

More knowledge on the composition of aqueous extracts of broiler litter amended for Water Soluble P (WSP) reduction would help to understand how amendments work. We measured pH, concentrations of Ca, Mg, Fe, Al, Cu, Mn, Zn, Molybdate Reactive P (MRP), and Dissolved Unreactive P (DUP) in water extracts of broiler litter treated with aluminum sulfate (ALS), ferrous sulfate (FES), ferric chloride (FEC), and gypsum (GYP) at 0, 5, 15, and 25% w/w. In order to study the effects of acidification, the same properties were measured in aqueous extracts of broiler litter suspensions that were titrated to end-points 3, 4, or 6 with 0.5N HCl. Concentrations of MRP, DUP, Ca and Mg, were 61%, 53%, 3.8 times, and 2.6 times greater in extracts from suspensions acidified to pH 6 than at the original pH of 8.9. ALS, FES, and FEC reduced pH, and showed similar effects on WSP concentrations, which were greater than with GYP. The magnitude of the reductions in WSP by ALS, FES, and FEC is uncertain because the actual amount of WSP immobilized cannot be determined. This is because of two opposite effects: 1) Through adsorption, soluble aluminum and iron remove phosphates from solution, and 2) Through acidification, iron and aluminum compounds release phosphates to solution.     

Interaction of soil pH and phosphorus efficacy: Long-term effects of P fertilizer and lime applications on wheat,barley, and sugar beet

Phosphorus (P), a plant macronutrient, must be adequately supplied for crop growth. In Germany, many soils are high in plant-available P; specifically in arable farming, P fertilizer application has been reduced or even omitted in the last decade. Therefore, it is important to understand how long these soils can support sustainable crop production, and what concentrations of soil P are required for it. We analyzed a 36-year long-term field experiment regarding the effects of different P application and liming rates on plant growth and soil P concentrations with a crop rotation of sugar beet, wheat, and barley. Sugar beet reacted to low soil P and low soil pH levels more sensitively than wheat, which was not significantly affected by the long-term omitted P application. All three crop species showed adequate growth at soil P levels lower than the currently recommended levels, if low soil pH was optimized by liming. The increase in efficacy of soil and fertilizer P by reduced P application rates therefore requires the adaptation of the soil pH to a soil type-specific optimal level.     

Agro-industrial wastes as effective amendments for ecotoxicity reduction and soil health improvement in aided phytostabilization

Aided phytostabilization is a technology that uses metal tolerant plants and organic and/or inorganic amendments to reduce soil metal bioavailability, while improving soil health. Our objective was to determine the effects of the application of amendments [sheep manure (SHEEP), poultry litter (POULTRY), cow slurry (COW), and paper mill sludge mixed with poultry litter (PAPER)], together with the growth of a metallicolous Festuca rubra L. population, on (i) chemical and microbial indicators of soil health and (ii) soil ecotoxicity, during the aided phytostabilization of a Zn/Pb contaminated mine soil. Amendment application led to an increase in soil pH, organic matter content, and inorganic salts, resulting in a decrease in Pb and Zn CaCl₂-extractable concentrations in soil, which, in turn, contributed to lower ecotoxicity and a stimulation of plant growth and soil microbial communities. The factor most affecting the metal extractability was probably soil pH. POULTRY was the best amendment in terms of increasing plant growth, chlorophylls content, and soil microbial biomass and activity, but resulted in higher levels of phytoavailable Pb and Zn. SHEEP and PAPER were more effective at reducing metal CaCl₂-extractability and, consequently, led to lower values of metal accumulation in plant tissues, thereby reducing the risk of metals entering into the food chain. When combined with the application of organic amendments, the metallicolous F. rubra population studied here appears an excellent candidate for aided phytostabilization. Our results indicate that the application of organic amendments is essential for the short-term recovery of highly contaminated metalliferous soils during aided phytostabilization.     

Composition of Aqueous Extracts of Broiler Litter Treated with Aluminum Sulfate,Ferrous Sulfate,Ferric Chloride and Gypsum

More knowledge on the composition of aqueous extracts of broiler litter amended for Water Soluble P (WSP) reduction would help to understand how amendments work. We measured pH, concentrations of Ca, Mg, Fe, Al, Cu, Mn, Zn, Molybdate Reactive P (MRP), and Dissolved Unreactive P (DUP) in water extracts of broiler litter treated with aluminum sulfate (ALS), ferrous sulfate (FES), ferric chloride (FEC), and gypsum (GYP) at 0, 5, 15, and 25% w/w. In order to study the effects of acidification, the same properties were measured in aqueous extracts of broiler litter suspensions that were titrated to end-points 3, 4, or 6 with 0.5N HCl. Concentrations of MRP, DUP, Ca and Mg, were 61%, 53%, 3.8 times, and 2.6 times greater in extracts from suspensions acidified to pH 6 than at the original pH of 8.9. ALS, FES, and FEC reduced pH, and showed similar effects on WSP concentrations, which were greater than with GYP. The magnitude of the reductions in WSP by ALS, FES, and FEC is uncertain because the actual amount of WSP immobilized cannot be determined. This is because of two opposite effects: 1) Through adsorption, soluble aluminum and iron remove phosphates from solution, and 2) Through acidification, iron and aluminum compounds release phosphates to solution.     

Internal acid buffering in San Joaquin Valley fog drops and its influence on aerosol processing

Although several chemical pathways exist for S(IV) oxidation in fogs and clouds, many are self-limiting: as sulfuric acid is produced and the drop pH declines, the rates of these pathways also decline. Some of the acid that is produced can be buffered by uptake of gaseous ammonia. Additional internal buffering can result from protonation of weak and strong bases present in solution. Acid titrations of high pH fog samples (median pH=6.49) collected in California's San Joaquin Valley reveal the presence of considerable internal acid buffering. In samples collected at a rural location, the observed internal buffering could be nearly accounted for based on concentrations of ammonia and bicarbonate present in solution. In samples collected in the cities of Fresno and Bakersfield, however, significant additional, unexplained buffering was present over a pH range extending from approximately four to seven. The additional buffering was found to be associated with dissolved compounds in the fogwater. It could not be accounted for by measured concentrations of low molecular weight (C₁–C₃) carboxylic acids, S(IV), phosphate, or nitrophenols. The amount of unexplained buffering in individual fog samples was found to correlate strongly with the sum of sample acetate and formate concentrations, suggesting that unmeasured organic species may be important contributors. Simulation of a Bakersfield fog episode with and without the additional, unexplained buffering revealed a significant impact on the fog chemistry. When the additional buffering was included, the simulated fog pH remained 0.3–0.7 pH units higher and the amount of sulfate present after the fog evaporated was increased by 50%. Including the additional buffering in the model simulation did not affect fogwater nitrate concentrations and was found to slightly decrease ammonium concentrations. The magnitude of the buffering effect on aqueous sulfate production is sensitive to the amount of ozone present to oxidize S(IV) in these high pH fogs.     

Metal accumulation from contaminated food and its effect on growth of juvenile landsnails Helix engaddensis

Metal accumulation by juvenile landsnails, Helix engaddensis, and its effect on growth rate was studied over a 5-week period of exposure and 2 weeks of recovery. An artificial food contaminated with Cu (4-2500 microg x g(-1)), Cd (50-800 microg x g(-1)), Pb, and Zn (20-12500 microg x g(-1)) was used. During the 7 weeks of the experiment, mortality rates were 20, 27, 30, and 38% among snails fed Cu-, Pb-, Zn-, and Cd-contaminated food, respectively. According to the ability to inhibit growth, metals were found to have the following order: Cd > Zn > Cu = Pb. Inhibitory effects of dietary metals started to be significant from the third week of exposure on. Inhibition of growth by Pb and Cu was found to be reversible, and within the first week of recovery, snails erupted their aestivation and resumed feeding and growth to gain weights similar to those of the control groups. Snails fed Cd- or Zn-contaminated food failed to resume growth during the 2 weeks of recovery. This indicates that in the case of Cu and Pb, growth inhibition was mainly due to starvation due to food rejection and aestivation. On the other hand, growth inhibition caused by Cd and Zn may have been resulting from irreversible toxicity. Therefore, snails were assumed to be sensitive to Cd and Zn but tolerant to Cu and Pb. Accumulation of Cu and Pb was significant only at the highest concentrations. At low and medium concentrations, no signs of accumulation were observed, indicating regulation at these concentrations. Cd and Zn accumulation starts at low concentrations but became significant at medium and high levels indicating accumulation of these metals.     

Effect of copper on growth of an aquatic macrophyte,Elodea canadensis

Elodea canadensis has been proposed as a potential biomonitor due to its wide distribution and apparent ability to accumulate pollutants in aquatic ecosystems. We investigated the effects of copper sulfate on growth in E. canadensis to determine its effectiveness as a biomonitor of copper pollution in aquatic systems and whether growth is a suitable index of sub-lethal stress. Copper sulfate significantly slowed or stopped growth at all concentrations (low: 1 ppm, medium: 5 ppm, high: 10 ppm of copper sulfate) used. Final plant drymass was significantly lower in medium and high copper treatments compared with controls. E. canadensis appears to be very sensitive to copper levels, and may be useful as a biomonitor of copper levels in aquatic systems. However, its utility as a bioaccumulator may be limited, because we observed senescence of most leaves in all copper-treated plants following 4 weeks of treatment.     

Sulfur Toxicity and Media Capacity for H2S Removal in Biofilters Packed with a Natural or a Commercial Granular Medium

Abstract

Two types of media, a natural medium (wood chips) and a commercially engineered medium, were evaluated for sulfur inhibition and capacity for removal of hydrogen sulfide (H₂S). Sulfate was added artificially (40, 65, and 100 mg of S/g of medium) to test its effect on removal efficiency and the media. A humidified gas stream of 50 ppm by volume H₂S was passed through the media-packed columns, and effluent readings for H₂S at the outlet were measured continuously. The overall H₂S baseline removal efficiencies of the column packed with natural medium remained >95% over a 2-day period even with the accumulated sulfur species. Added sulfate at a concentration high enough to saturate the biofilter moisture phase did not appear to affect the H₂S removal process efficiency. The results of additional experiments with a commercial granular medium also demonstrated that the accumulation of amounts of sulfate sufficient enough to saturate the moisture phase of the medium did not have a significant effect on H₂S removal.

When the pH of the biofilter medium was lowered to 4, H₂S removal efficiency did drop to 36%. This work suggests that sulfate mass transfer through the moisture phase to the biofilm phase does not appear to inhibit H₂S removal rates in biofilters. Thus, performance degradation for odor-removing biofilters or H₂S breakthrough in field applications is probably caused by other consequences of high H₂S loading, such as sulfur precipitation.     

Comparison of spontaneous antibiotic resistance frequency of Salmonella Typhimurium growth in glucose amended continuous culture at slow and fast dilution rates

The objective of the study was to determine the frequency of spontaneous acquisition of resistance to select antibiotics by Salmonella Typhimurium (ST) when grown in glucose amended continuous flow culture at slow (D = 0.025 h(-1)) or fast (D = 0.27 h(-1)) dilution rates. The bacterium was grown in LB minimal medium (pH 6.25) containing no antibiotics. Upon achieving steady state, samples were plated to tryptic soy agar (TSA) alone or supplemented (per ml) with 2 and 16 microg oxytetracycline, 4 and 16 microg tetracycline, 2 and 64 microg kanamycin, and 0.25 and 2 microg enrofloxacin. Regardless of growth rate, CFU of resistant ST from the TSA containing antibiotics was less than 2 x 10(1) except for 2 microg kanamycin and 0.25 microg enrofloxacin treatments (higher than 1 x 10(9) and 4 x 10(7) CFU of resistant ST for trials 1 and 2, respectively). Frequency of recovering resistant ST from the TSA containing the higher antibiotic concentrations was less than 1 in 10(9) for all antibiotics, but was higher on the media containing 2 microg kanamycin and 0.25 microg enrofloxacin at both slow and fast growth rates. In general, minimal susceptibility differences were detected for isolates from slow and fast dilution rates.     

Effects of aluminium in acid streams on growth and sporulation of aquatic hyphomycetes

We investigated, by field and laboratory experiments, the effects of aluminium in an acid stream (pH 5.0) on the growth and sporulation of aquatic hyphomycete fungi which degrade organic litter. The stream water had monomeric aluminium (Al(m)) concentrations of 9.1-13.4 microm - fifty times higher than a nearby circumneutral stream. Alder leaves submersed in the stream accumulated Al, most of which was tightly bound. Growth rates of four species of aquatic hyphomycetes were altered by inclusion of Al(m) in the culture medium. On a polypectate substrate, and on low-phosphate medium with glucose, growth rates increased significantly. On a low-nutrient substrate of homogenized alder leaves, growth rates were inhibited by aluminium. The pattern of mycelial growth was found to be different on a polypectate medium including Al(m), compared with a control without aluminium. There was a significant increase in hyphal radial growth and a decrease in the hyphal growth unit. The effect resembled the growth of a starved fungal colony. Treatment with Al(m) decreased pectinase production by the four fungal species tested. The capacity of these species to sporulate was reduced by flooding culture plates with Al(m) solution. These deleterious metabolic effects were most severe in isolates taken from circumneutral streams and less marked, though significant, in species originating from acid streams.