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Xiuping Jiang

Publications and source records attributed to Xiuping Jiang.

16 recordsLinked to original sources

Analysis of fecal microbial flora for antibiotic resistance in ceftiofur-treated calves.

To evaluate the impact of ceftiofur treatment in calves on fecal shedding of ceftriaxone-resistant bacteria, 3 female Holstein dairy calves were treated by intramuscular injection with EXCENEL RTU (ceftiofur hydrochloride, Pharmacia and Upjohn) at a therapeutic dosage of 2.2 mg/kg/day for 5 consecutive days following label directions. Three untreated calves were housed separately and served as controls. One to 3 days following the initial administration of ceftiofur, there was a 14% and 2% increase of fecal bacteria resistant to 16 and 64 microg ceftriaxone/mL, respectively. This response remained unchanged from days 6 to 13, and increased resistance was seen at day 17. Randomly selected isolates of gram-positive and gram-negative bacteria with elevated resistance to ceftriaxone (minimal inhibitory concentration (MIC) >or=64 microg ceftriaxone/mL) were isolated from calf feces and identified. In vitro conjugation experiments revealed that both the ceftriaxone-resistance gene bla (CMY-2) and class 1 integron were transferred from two bacterial species to Salmonella spp. at a frequency of 10(7) to 10(5). MIC data revealed that Salmonella transconjugants acquired either reduced susceptibility or resistance to ceftriaxone as well as to multiple antibiotics. This genetic transfer occurred both within and between genera. Treatment of calves with therapeutic dosages of ceftiofur can significantly increase for at least 17 days following the initial treatment the fecal excretion of ceftriaxone-resistant bacteria, including Salmonella species.

Animals↗

Occurrence of ceftriaxone-resistant commensal bacteria on a dairy farm and a poultry farm.

Approximately 40 samples of animal feces, drinking water, feed, bedding, pine wood shavings, compost, and manure slurry were collected from two animal research farms (one dairy and one poultry) and analyzed for ceftriaxone-resistant bacteria. Our study revealed that the total percentage of aerobic bacteria with reduced susceptibility to ceftriaxone (minimal inhibitory concentration (MIC) > or = 16 micro g/mL) ranged from 0.9% to 10.8% in dairy feces and from 0.05% to 3.93% in chicken feces. The percentages of ceftriaxone-resistant bacteria (MIC > or = 64 micro g/mL) were in the range of 0.01% - 2.3% in dairy feces and 0.01% - 0.79% in chicken feces. Environmental samples contained a wide range of ceftriaxone-resistant bacterial populations. Among those environmental samples, fresh pine wood shavings used as chicken bedding contained the highest percentages (41.5%) of ceftriaxone-resistant bacteria, as determined by a plating method. A total of 105 ceftriaxone-resistant (MIC > or = 128 micro g/mL) bacterial isolates were isolated from the above samples and tested for resistance to nine antibiotics: ampicillin, ceftriaxone, streptomycin, kanamycin, gentamicin, chloramphenicol, tetracycline, ciprofloxacin, and nalidixic acid. The most prevalent resistance pattern (34.3%) among isolates included resistance to all nine antibiotics. Results from this study suggest that ceftriaxone-resistant bacteria exist in farm environments, and the ceftriaxone resistance was frequently associated with resistance to multiple antibiotics. Environmental sources such as pine wood shavings used as bedding can be a potential reservoir for transmitting the multidrug-resistant bacteria.

Agriculture↗

Carbon nanotubes for immunomagnetic separation of Escherichia coli O157: H7.

Bovine serum albumin-functionalized multiple-walled carbon nanotubes with encapsulated ferromagnetic elements were conjugated with pathogen-specific antibody, and the conjugate was evaluated for immunomagnetic separation of Escherichia coli O157:H7 in pure and mixed (with Salmonella Typhimurium) cultures.

Coated Materials, Biocompatible↗

Improvements in the cell-free production of functional antibodies using cell extract from protease-deficient Escherichia coli mutant.

Expression of a functional antibody fragment (Fab) using an Escherichia coli cell-free expression system has been reported previously [Jiang et al., FEBS Lett., 514, 290-294 (2002)]. The low yield of the synthesized antibody, however, limits the usefulness of the cell-free expression system, partly due to the degradation of product by endogenous proteases from the E. coli extract. To determine which proteases are responsible for the degradation, we compared the expression of a 6D9 Fab fragment under conditions whereby several protease inhibitors were added into the cell-free system. The addition of serine protease inhibitor increased the amount of the Fab fragment, indicating that serine proteases caused the antibody degradation. Therefore, several serine protease-deficient mutants of E. coli BW25113 were constructed by targeted homologous recombination. The use of extract from a double protease-deficient mutant (DeltadegP-ompT) significantly increased the amount and antigen-binding activities of an anti-HSA scFv and a 6D9 Fab fragment. These results suggest that the DegP- and OmpT-deleted mutant is a useful source of S30 extract for the production or screening of antibodies using the cell-free expression system.

Antibody Formation↗

Persistence of Salmonella enterica serovar typhimurium on lettuce and parsley and in soils on which they were grown in fields treated with contaminated manure composts or irrigation water.

There are many sources of pathogen contamination of vegetable crops in the field that include manure used as fertilizer and irrigation water. An avirulent strain of Salmonella enterica serovar Typhimurium was added to three different types of composts-PM-5 (poultry manure compost), 338 (dairy manure compost), and NVIRO-4 (alkaline stabilized dairy manure compost)-and irrigation water at 10(7) colony forming units (cfu)/g and 10(5) cfu/mL, respectively, to determine under field conditions the persistence of salmonellae in soils treated with these composts or irrigation water, and also on leaf lettuce and parsley grown on such treated soil. Contaminated compost was applied to soil in the field as a strip at a rate of 4.5 metric tons/hectare on the day before lettuce and parsley seedlings were transplanted. Contaminated irrigation water was applied only once on the plants at the rate of 2 liters per plot on the same day after the seedlings were transplanted. Twenty-five plots, each measuring 1.8 x 4.6 meters, were used for each crop, with five treatments (one without compost, three with each of the three composts, and one without compost but applied with contaminated water) and five replication plots for each treatment. Salmonella persisted for 161 and up to 231 days in soils amended with contaminated composts on which lettuce and parsley, respectively, were grown, and was detected for up to 63 days and 231 days on lettuce and parsley, respectively. The type of contaminated compost had minimal effect on the persistence of S. Typhimurium in soil. Occurrence of Salmonella on vegetables and survival in soil on which these vegetables were grown, irrespective of source of contamination through irrigation water or compost, were similar, suggesting both contaminated manure compost and irrigation water can play important roles in contaminating soil and vegetables with Salmonella for an extended period of time.

Colony Count, Microbial↗

Fate of Salmonella enterica serovar Typhimurium on carrots and radishes grown in fields treated with contaminated manure composts or irrigation water.

Three different types of compost, PM-5 (poultry manure compost), 338 (dairy cattle manure compost), and NVIRO-4 (alkaline-pH-stabilized dairy cattle manure compost), and irrigation water were inoculated with an avirulent strain of Salmonella enterica serovar Typhimurium at 10(7) CFU g(-1) and 10(5) CFU ml(-1), respectively, to determine the persistence of salmonellae in soils containing these composts, in irrigation water, and also on carrots and radishes grown in these contaminated soils. A split-plot block design plan was used for each crop, with five treatments (one without compost, three with each of the three composts, and one without compost but with contaminated water applied) and five replicates for a total of 25 plots for each crop, with each plot measuring 1.8 x 4.6 m. Salmonellae persisted for an extended period of time, with the bacteria surviving in soil samples for 203 to 231 days, and were detected after seeds were sown for 84 and 203 days on radishes and carrots, respectively. Salmonella survival was greatest in soil amended with poultry compost and least in soil containing alkaline-pH-stabilized dairy cattle manure compost. Survival profiles of Salmonella on vegetables and soil samples contaminated by irrigation water were similar to those observed when contamination occurred through compost. Hence, both contaminated manure compost and irrigation water can play an important role in contaminating soil and root vegetables with salmonellae for several months.

Agriculture↗

Fate of Escherichia coli O157:H7 in manure compost-amended soil and on carrots and onions grown in an environmentally controlled growth chamber.

Studies were done to determine the fate of Escherichia coli O157:H7 in manure compost-amended soil and on carrots and green onions grown in an environmentally controlled growth chamber. Commercial dairy cattle manure compost was inoculated with a five-strain mixture of green fluorescent protein-labeled E. coli O157:H7 at 10(7) CFU g(-1) and mixed with unsterilized Tifton sandy loam soil at a ratio of 1:5. Baby carrot or green onion seedlings were planted into the manure compost-amended soil in pots, and soil samples surrounding the plant, edible carrot roots and onion bulb samples, and soil immediately beneath the roots were assayed for E. coli O157:H7 in triplicate at weekly intervals for the first 4 weeks, and every 2 weeks for the remainder of the plant growth cycle (up to 3 months). E. coli O157:H7 cell numbers decreased within 64 days by 3 log CFU/g in soil and soil beneath the roots of green onions and by more than 2 log CFU/g on onions. E. coli O157:H7 survived better during the production of carrots, with a 2.3-log CFU/g reduction in soil and a 1.7-log CFU/g reduction on carrots within 84 days. These results indicate that the type of plant grown is an important factor influencing the survival of E. coli O157:H7 both on the vegetable and in the soil in which the vegetable is grown.

Agriculture↗

Persistence of enterohemorrhagic Escherichia coli O157:H7 in soil and on leaf lettuce and parsley grown in fields treated with contaminated manure composts or irrigation water.

Outbreaks of enterohemorrhagic Escherichia coli O157:H7 infections associated with lettuce and other leaf crops have occurred with increasing frequency in recent years. Contaminated manure and polluted irrigation water are probable vehicles for the pathogen in many outbreaks. In this study, the occurrence and persistence of E. coli O157:H7 in soil fertilized with contaminated poultry or bovine manure composts or treated with contaminated irrigation water and on lettuce and parsley grown on these soils under natural environmental conditions was determined. Twenty-five plots, each 1.8 by 4.6 m, were used for each crop, with five treatments (one without compost, three with each of the three composts, and one without compost but treated with contaminated water) and five replication plots for each treatment. Three different types of compost, PM-5 (poultry manure compost), 338 (dairy manure compost), and NVIRO-4 (alkaline-stabilized dairy manure compost), and irrigation water were inoculated with an avirulent strain of E. coli O157:H7. Pathogen concentrations were 10(7) CFU/g of compost and 10(5) CFU/ml of water. Contaminated compost was applied to soil in the field as a strip at 4.5 metric tons per hectare on the day before lettuce and parsley seedlings were transplanted in late October 2002. Contaminated irrigation water was applied only once on the plants as a treatment in five plots for each crop at the rate of 2 liters per plot 3 weeks after the seedlings were transplanted. E. coli O157:H7 persisted for 154 to 217 days in soils amended with contaminated composts and was detected on lettuce and parsley for up to 77 and 177 days, respectively, after seedlings were planted. Very little difference was observed in E. coli O157:H7 persistence based on compost type alone. E. coli O157:H7 persisted longer (by > 60 days) in soil covered with parsley plants than in soil from lettuce plots, which were bare after lettuce was harvested. In all cases, E. coli O157:H7 in soil, regardless of source or crop type, persisted for > 5 months after application of contaminated compost or irrigation water.

Disease Outbreaks↗

Fate of Listeria monocytogenes in bovine manure-amended soil.

The survival and growth of Listeria monocytogenes in soil amended with bovine manure was studied under different environmental conditions of temperature, nutrients, and soil microflora. Autoclaved soil was compared with unautoclaved soil for assessing the influence of competitive soil microflora on the survival of L. monocytogenes. Initial L. monocytogenes cell numbers of 5 to 6 log CFU/g survived for up to 43, 43, and 14 days in manure-amended autoclaved soil at 5, 15, and 21 degrees C, respectively. In manure-amended unautoclaved soil, the pathogen was detectable for up to 43, 21, and 21 days at 5, 15, and 21 degrees C, respectively. L. monocytogenes was inactivated more rapidly in autoclaved soil amended with manure at a manure/soil ratio of 1:10 than in the more dilute (1:100) manure in soil samples at both 15 and 21 degrees C. However, in manure-amended unautoclaved soil, L. monocytogenes survived longer in samples with ratios of 1:10 than in the more dilute (1:100) manure-amended soil. The persistence of L. monocytogenes for several weeks in manure-amended soil suggests listeriae could be transmitted through soil to fresh produce or to shoes, clothing, and hands of field workers, especially during the cold months.

Antibiosis↗

Fate of Escherichia coli O157:H7 during composting of bovine manure in a laboratory-scale bioreactor.

Inactivation profiles of Escherichia coli O157:H7 in inoculated bovine manure-based compost ingredients were determined by composting these ingredients in a bioreactor under controlled conditions. A 15-liter bioreactor was constructed to determine the fate of E. coli O157:H7 and changes in pH, moisture content, temperature, and aerobic mesophilic and thermophilic bacterial counts during composting. Fresh cow manure, wheat straw, cottonseed meal, and ammonium sulfate were combined to obtain a moisture content of ca. 60% and a carbon/nitrogen ratio of 29:1. The compost ingredients were held in the bioreactor at a constant external temperature of 21 or 50 degrees C. Self-heating of the ingredients due to microbial activity occurred during composting, with stratified temperatures occurring within the bioreactor. At an external temperature of 21 degrees C, self-heating occurred for 0 to 3 days, depending on the location within the bioreactor. E. coli O157:H7 populations increased by 1 to 2 log10 CFU/g during the initial 24 h of composting and decreased by ca. 3.5 log10 CFU/g near the bottom of the bioreactor and by ca. 2 log10 CFU/g near the middle and at the top during 36 days of composting. At an external temperature of 50 degrees C. E. coli O157:H7 was inactivated rapidly (by ca. 4.9 log10 CFU/g at the top of the bioreactor, by 4.0 log10 CFU/g near the middle, and by 5.9 log10 CFU/g near the bottom) within 24 h of composting. When inoculated at an initial level of ca. 10(7) CFU/g. E. coli O157:H7 survived for 7 days but not for 14 days at all three sampling locations, as indicated by either direct plating or enrichment culture. At the top of the bioreactor a relatively constant moisture content of 60% was maintained, whereas the moisture content near the bottom decreased steadily to 37 to 45% over 14 days of composting. The pH of the composting mixture decreased to ca. 6 within 1 to 3 days and subsequently increased to 8 to 9. Results obtained in this study indicate that large populations (10(4) to 10(7) CFU/g) of E coli O157:H7 survived for 36 days during composting in a bioreactor at an external temperature of 21 degrees C but were inactivated to undetectable levels after 7 to 14 days when the external temperature of the bioreactor was 50 degrees C. Hence, manure contaminated with large populations (e.g., 10(7) CFU/g) of E. coli O157:H7 should be composted for more than 1 week, and preferably for 2 weeks, when held at a minimum temperature of 50 degrees C.

Animals↗

Thermal inactivation of Escherichia coli O157:H7 in cow manure compost.

Rates of inactivation of a five-strain mixture of green fluorescent protein-labeled Escherichia coli O157:H7 in autoclaved and unautoclaved commercial cow manure compost with a moisture content of ca. 38% were determined at temperatures of 50, 55, 60, 65, and 70 degrees C. Trypticase soy agar with ampicillin was determined to be the best medium for the enumeration of heat-injured and uninjured cells of green fluorescent protein-labeled E. coli O157:H7. The results obtained in this study revealed that in autoclaved compost, E. coli O157:H7 reductions of ca. 4 log CFU/g occurred within 8 h, 3 h, 15 min, 2 min, and < 1 min at 50, 55, 60, 65, and 70 degrees C, respectively. At 65 and 70 degrees C, considerably less time was required to kill the pathogen in unautoclaved compost than in autoclaved compost. Decimal reduction times (D-values) for autoclaved compost at 50, 55, 60, 65, and 70 degrees C were 137, 50.3, 4.1, 1.8, and 0.93 min, respectively, and D-values for unautoclaved compost at 50, 55, and 60 degrees C were 135, 35.4, and 3.9 min, respectively. Considerable tailing was observed for inactivation curves, especially at 60, 65, and 70 degrees C. These results are useful for identifying composting conditions that will reduce the risk of the transmission of E. coli O157:H7 to foods produced in the presence of animal fecal waste.

Animals↗

Fate of Escherichia coli O157:H7 in manure-amended soil.

Escherichia coli O157:H7 cells survived for up to 77, >226, and 231 days in manure-amended autoclaved soil held at 5, 15, and 21 degrees C, respectively. Pathogen populations declined more rapidly in manure-amended unautoclaved soil under the same conditions, likely due to antagonistic interactions with indigenous soil microorganisms. E. coli O157:H7 cells were inactivated more rapidly in both autoclaved and unautoclaved soils amended with manure at a ratio of 1 part manure to 10 parts soil at 15 and 21 degrees C than in soil samples containing dilute amounts of manure. The manure-to-soil ratio, soil temperature, and indigenous microorganisms of the soil appear to be contributory factors to the pathogen's survival in manure-amended soil.

Antibiosis↗

Reduction of protein degradation by use of protease-deficient mutants in cell-free protein synthesis system of Escherichia coli.

In an Escherichia coli in vitro transcription/translation system, the degradation of produced proteins is often caused by endogenous proteases from E. coli extracts. To reduce the extent of this degradation, several extracts were prepared from E. coli mutants that genetically lacked DegP, OmpT, or Lon proteases. Then, these extracts were used with 14C-leucine in a system for synthesizing single-chain Fv against gp120 (anti-gp120), and phospholipase D (PLD) of Streptomyces antibioticus. The proteins synthesized in vitro were analyzed by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) and autoradiography. The use of extracts from mutants that were deficient in the structural genes encoding OmpT and Lon markedly repressed the degradation of anti-gp120. Similarly, extracts from degP- and ompT-deleted mutants were able to significantly stabilize in vitro-synthesized PLD, which otherwise disappeared within 30 min. Such protease-deficient mutants were suggested to be useful for preventing the degradation of heterologous proteins in in vitro systems.

Journal Article↗

Optimizing enrichment culture conditions for detecting Helicobacter pylori in foods.

The survival and growth of Helicobacter pylori under enrichment conditions in fresh, autoclaved and irradiated ground beef were determined. H. pylori grew in autoclaved ground beef at 37 degrees C under microaerobic conditions in brain heart infusion broth with 7% horse serum at pH 7.3 after 3 to 7 days of lag time but did not grow within 7 days in irradiated (10 kGy) ground beef under the same enrichment conditions. Adjustment of the enrichment broth to pH 5.5 enabled the growth (ca. 2 log10 CFU/ml) of H. pylori within 7 days in the presence of irradiated ground beef and the prolific growth (ca. 3 to 4 log10 CFU/ml) of H. pylori within 3 days in the presence of autoclaved beef. H. pylori in fresh ground beef could not be isolated from enrichment media with antibiotics; however. H. pylori ureA could be detected by polymerase chain reaction (PCR) in such enrichment media after 1 to 3 days of incubation at 37 degrees C. The addition of supplements, i.e., 0.3% mucin, 0.05% ferrous sulfate, and 0.05% sodium pyruvate or 0.008 M urea, or the adjustment of the enrichment broth pH to 5.5 or 4.5 enabled the detection of H. pylori ureA in enrichment media incubated for 1, 2, 3, and/or 7 days at 37 degrees C. H. pylori in sterile milk refrigerated at 4 degrees C at an initial level of 10(6) CFU/ml was inactivated to an undetectable level within 6 days; however, H. pylori was not detected either by a PCR assay or by the plating of enrichment cultures of 120 raw bovine milk samples.

Animals↗