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1.
Poult Sci ; 101(3): 101688, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35104730

ABSTRACT

Reduction of Salmonella on poultry carcasses is one way to prevent salmonellosis. The purpose of this research was to evaluate the effects of subzero saline chilling (SSC) with/without hot water spray (HWS) on broiler carcasses prior to chilling for bacterial reduction. Eviscerated broiler carcasses were subjected to water immersion chilling (WIC, 0% NaCl/0.5°C) or SSC (4% NaCl/-2.41°C) with/without prior HWS at 71°C for 1 min. Broiler carcasses in SSC were chilled faster than those in WIC, regardless of HWS. The combination of HWS and SSC resulted in the best reduction of mesophilic aerobic bacteria, Escherichia coli, and total coliforms on the carcasses over the WIC, SSC, and HWS/WIC. No Salmonella was detected on the carcasses in SSC and HWS/SSC while Salmonella positive was observed on the carcasses chilled in WIC and HWS/WIC. A trace of Gram-negative genus was detected on carcasses in HWS/SSC while many other microbiomes were observed on those in WIC, SSC, and HWS/WIC when quantitative microbiota profiles of 16S rRNA gene sequences were evaluated. Based on these results, chilling of broiler carcasses in 4% NaCl/-2.41°C after HWS at 71°C for 1 min significantly reduced carcass chilling time and bacterial contamination over the control chilling.


Subject(s)
Food Handling , Meat , Animals , Chickens/microbiology , Cold Temperature , Colony Count, Microbial/veterinary , Decontamination , Food Handling/methods , Food Microbiology , Meat/analysis , RNA, Ribosomal, 16S , Water
2.
Poult Sci ; 98(12): 6954-6963, 2019 Dec 01.
Article in English | MEDLINE | ID: mdl-31504943

ABSTRACT

The present study evaluated the efficacy of ethanol treatment (0, 30, 50, or 70%) alone or in combination with ultrasound (37 kHz, 380 W) for the reduction of natural indigenous mesophilic aerobic bacteria (MAB), coliforms, and inoculated Salmonella Typhimurium on chicken skin. Bacterial cells with loose, intermediate, or tight attachment to chicken skin were recovered by shaking in an incubator (200 rpm) for 5 min, stomaching for 1 min, or blending for 1 min, respectively. Chicken skins were inoculated with a suspension (7 log CFU/mL) of S. Typhimurium. Ethanol reduced the number of MAB, coliforms, and S. Typhimurium on the chicken skin in a concentration-dependent manner, whereas ultrasound treatment without ethanol was ineffective. A combination of 70% ethanol with ultrasound treatment was the most effective in reducing S. Typhimurium populations with loose, intermediate, and tight attachment (reduction by 2.86 log CFU/g, 2.49 log CFU/g, and 1.63 log CFU/g, respectively). However, chicken skin treated with 50% ethanol alone or with a combination of >50% ethanol and ultrasound showed significant changes in Hunter color values (a* and b*) and texture (shear force) (P > 0.05). On the other hand, a combination of 30% ethanol and ultrasound yielded the best results, leading to a reduction of S. Typhimurium by a >1.0 log CFU/g, but did not alter the color or texture of chicken skin. Thus, a combination of 30% ethanol and ultrasound appears to be the optimum treatment for reduction of microbial contamination in production and distribution of skin-on chicken products, and enhance poultry safety without decreasing food quality.


Subject(s)
Ethanol/pharmacology , Food Handling/methods , Food Microbiology , Salmonella typhimurium/drug effects , Skin/microbiology , Animals , Bacteria, Aerobic , Chickens/microbiology , Food Quality
3.
Poult Sci ; 98(3): 1539-1544, 2019 Mar 01.
Article in English | MEDLINE | ID: mdl-30169808

ABSTRACT

This study was conducted to evaluate antilisterial activity in deli-style turkey using one of the inhibitors: hop α- or ß-acids at 5 ppm, potassium acetate/potassium diacetate (PAPD) at 0.5%, their combinations, potassium lactate/sodium diacetate (PLSD) at 2.5% for positive control, and ethanol at 5 ppm for negative control. Deli-style turkey was formulated and manufactured traditionally. To simulate Listeria contamination in processing plants, the deli turkey was sliced in <24 h of production, inoculated with Listeria monocytogenes (2 to 3 log CFU/g), and vacuum-stored at 4 or 7°C for 60 d. To simulate Listeria contamination in grocery stores, the deli turkey was vacuum-stored for 30 and 60 d prior to slicing, inoculation, and aerobic storage at 4 or 7°C for 10 d. Physicochemical properties of the deli turkey were not significantly different among treatments (P > 0.05). Addition of hop acids at 5 ppm did not inhibit Listeria in deli meat during 60 d of vacuum-storage at 4 or 7°C, whereas organic acids and hop acids/PAPD significantly inhibited Listeria (P < 0.05), with the best inhibition observed for hop ß-acids/PAPD at 7°C/60 d. During 10 d of aerobic storage at 4°C, hop acid/PAPD, PLSD, or PAPD showed listeristatic effects, whereas hop acids alone allowed Listeria to grow by 0.9 CFU/g. During 10 d of aerobic storage at 7°C, hop acid/PAPD, PLSD, and PAPD significantly reduced Listeria populations than hop α-acids, ß-acids, and no-inhibitor control (P < 0.05). These results indicated the combination of hop ß-acids/PAPD provides more effective inhibition than any single addition of hop acids and PLSD (P < 0.05) at 7°C/60 d in vacuum storage, with intermediate inhibition observed for PAPD and α-acids/PAPD.


Subject(s)
Food Preservatives/pharmacology , Humulus/chemistry , Listeria monocytogenes/drug effects , Meat Products/microbiology , Acetates/chemistry , Animals , Ethanol/chemistry , Food Microbiology , Food Storage , Turkeys
4.
Poult Sci ; 97(6): 2207-2210, 2018 Jun 01.
Article in English | MEDLINE | ID: mdl-29762788

ABSTRACT

Chemical components of hop resins effectively inhibit the growth of L. monocytogenes in microbiological culture media. This study was conducted to investigate antilisterial activities of hop α- and ß-acid in turkey slurry. Turkey slurries were inoculated with L. monocytogenes, formulated with hop α- or ß-acid from 0 to 1,000 ppm, and incubated at 37°C for 24 h or at 7°C for 12 days. During storage at 37°C for 24 h, L. monocytogenes populations were reduced from 2.40 log CFU/g to non-detectable (<1 log CFU/g) in α-acid at ≥750 ppm and ß-acid at 1,000 ppm, whereas the control (0 ppm) allowed the pathogen to grow to 8.0 log CFU/g. During storage at 7°C for 12 d, the slurry treated with α-acid at ≥100 ppm and ß-acid at ≥500 ppm showed listeristatic effects, while listericidal effects were observed in the slurries at 1,000 ppm, regardless of hop acid type. Hop α-acid ≤ 50 ppm and ß-acid ≤ 100 ppm failed to inhibit L. monocytogenes, and the pattern of bacterial growth was similar to that of control with no significant difference (P > 0.05). Based on these results, the concentration of α-acid > 100 ppm or ß-acid > 500 ppm is minimally required to inhibit L. monocytogenes when turkey batters are formulated with hop acids as a single antilisterial agent prior to cooking and storage at 7°C.


Subject(s)
Anti-Infective Agents/pharmacology , Food Handling/methods , Food Microbiology , Humulus/chemistry , Listeria monocytogenes/drug effects , Poultry Products/microbiology , Animals , Temperature , Turkeys
5.
Poult Sci ; 97(5): 1681-1688, 2018 May 01.
Article in English | MEDLINE | ID: mdl-29244088

ABSTRACT

This study investigated bactericidal activity of 0.05 to 0.50% calcium oxide (CaO) against planktonic cells in tryptic soy broth (TSB) and biofilms of Listeria monocytogenes on eggshell surfaces. The bactericidal activity of CaO against planktonic cells and biofilms of L. monocytogens significantly (P < 0.05) increased log reductions with increasing concentrations of CaO. Exposure to 0.05 to 0.50% CaO for one min reduced planktonic cells in TSB cell suspensions by 0.47 to 3.86 log10CFU/mL and biofilm cells on the shell surfaces by 0.14 to 2.32 log10CFU/cm2. The Hunter colors of eggshells ("L" for lightness, "a" for redness, and "b" for yellowness), shell thickness (puncture force), and sensory quality (egg taste and yolk color) were not changed by 0.05 to 0.50% CaO treatment. The nonlinear Weibull model was used to calculate CR = 3 values as the CaO concentration of 3 log (99.9%) reduction for planktonic cells (R2 = 0.96, RMSE = 0.26) and biofilms (R2 = 0.95, RMSE = 0.18) of L. monocytogens. The CR = 3 value, 0.31% CaO for planktonic cells, was significantly (P < 0.05) lower than 0.57% CaO for biofilms. CaO could be an alternative disinfectant to reduce planktonic cells and biofilms L. monocytogenes on eggshell surface in egg processing plants.


Subject(s)
Biofilms/drug effects , Calcium Compounds/pharmacology , Disinfectants/pharmacology , Egg Shell/microbiology , Food Microbiology/methods , Listeria monocytogenes/drug effects , Listeria monocytogenes/physiology , Oxides/pharmacology , Animals , Chickens , Pectinidae/chemistry , Powders
6.
Lett Appl Microbiol ; 64(1): 27-34, 2017 Jan.
Article in English | MEDLINE | ID: mdl-27747902

ABSTRACT

Microbiological quality of 206 raw ready-to-eat seafood samples was investigated according to species (gizzard shad, halibut, rockfish, tuna, oyster and squid) and distribution channels (fishery, hyper and online market). Enumeration of aerobic plate count and total coliforms (TC) and pathogenic bacteria (Bacillus cereus, Staphylococcus aureus and Vibrio parahaemolyticus) was performed, and level of microbiological quality was classified into four groups: satisfactory, acceptable, unsatisfactory and unacceptable. Qualitative analysis was also performed for Escherichia coli and eight foodborne pathogens (B. cereus, E. coli O157:H7, Listeria monocytogenes, Salmonella spp., S. aureus, Vibrio cholerae, V. parahaemolyticus, and Vibrio vulnificus). Raw ready-to-eat seafood products revealed 0·5% at an unsatisfactory level and 4·9% at an unacceptable level due to ≥4 log CFU g-1 of TC in squid and ≥3 log CFU g-1 of V. parahaemolyticus in gizzard shad respectively. Gizzard shad was shown to be potentially hazardous, as its sashimi is eaten with its skin attached. Bacillus cereus, E. coli, S. aureus, V. parahaemolyticus and V. vulnificus were qualitatively detected. Samples from the fishery market showed higher detection rate especially in V. parahaemolyticus (21·6%) and V. vulnificus (1·7%) which indicates the need to improve microbiological safety of raw ready-to-eat seafood products in fishery market. SIGNIFICANCE AND IMPACT OF THE STUDY: Raw ready-to-eat seafood products like sashimi can be easily contaminated with various bacteria from aquatic environments and human reservoirs, which subsequently bring about a risk in food poisoning due to no heating process before consumption. The results of this study provide comprehensive microbiological data on various species of raw ready-to-eat seafood from various distribution channels. It may contribute to establish reasonable standard and effective strategies to ensure a good microbiological quality of raw ready-to-eat seafood for the safety of meals, like sashimi and sushi.


Subject(s)
Fish Products/microbiology , Food Contamination/analysis , Food Microbiology , Food Safety , Foodborne Diseases/microbiology , Seafood/microbiology , Shellfish/microbiology , Animals , Bacillus cereus/isolation & purification , Colony Count, Microbial , Escherichia coli O157/isolation & purification , Fisheries , Humans , Listeria monocytogenes/isolation & purification , Ostreidae/microbiology , Salmonella/isolation & purification , Staphylococcal Infections , Staphylococcus aureus/isolation & purification , Vibrio parahaemolyticus/isolation & purification , Vibrio vulnificus/isolation & purification
7.
Poult Sci ; 94(12): 3034-9, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26467007

ABSTRACT

This research was conducted to quantify bacterial populations after swabbing or stomaching, followed by grinding the swabbed or stomached broiler skins. For each of 3 replications, 3 eviscerated broilers were randomly taken from a processing line in a local broiler processing plant. Ten swabs and 10 stomachs per bird were conducted on the left- and the right-side skins (10×7 cm), respectively, which were then finally ground. Results indicated that mesophilic aerobic bacteria (MAB) in the first swabbed sample were significantly lower than those in the first stomached sample (P<0.05), with no difference seen for the remaining sampling times (P>0.05). During 10 swabbings followed by final grinding, 8, 9, and 83% of MAB were detected after the first swabbing, after the second through 10th swabbings, and after final grinding of the skin, respectively. During 10 stomachings followed by the final grinding, 17, 18, and 65% of MAB were detected after the first stomaching, after the second through 10th stomachings, and after final grinding of the skin, respectively. Escherichia coli (E. coli) and coliforms were significantly higher in the first stomaching than those in the first swabbing (P<0.05), with no difference seen between the 2 sampling methods for the rest sampling times (P>0.05). Populations of E. coli and coliforms decreased step-wisely from the highest after grinding to the intermediate after first and second sampling, and to the least after 10th sampling (P<0.05), regardless of swabbing or grinding. In this study, less than 35% of MAB seemed loosely associated in the skin of eviscerated broiler, whereas more than 65% of MAB looked tightly associated, which were not recovered by stomaching or swabbing even 10 times but were recovered by grinding the skin.


Subject(s)
Bacteria/isolation & purification , Food Contamination/analysis , Food Handling/methods , Meat/microbiology , Skin/microbiology , Animals , Chickens , Enterobacteriaceae/isolation & purification , Escherichia coli/isolation & purification , Food Microbiology
8.
Poult Sci ; 93(1): 200-7, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24570440

ABSTRACT

The inhibitory effect of chlorine (50, 100, and 200 mg/kg) was investigated with and without UV radiation (300 mW·s/cm(2)) for the growth of Listeria monocytogenes in chicken breast meat. Using a polynomial model, predictive growth models were also developed as a function of chlorine concentration, UV exposure, and storage temperature (4, 10, and 15°C). A maximum L. monocytogenes reduction (0.8 log cfu, cfu/g) was obtained when combining chlorine at 200 mg/kg and UV at 300 mW·s/cm(2), and a maximum synergistic effect (0.4 log cfu/g) was observed when using chlorine at 100 mg/kg and UV at 300 mW·s/cm(2). Primary models developed for specific growth rate and lag time showed a good fitness (R(2) > 0.91), as determined by the reparameterized Gompertz equation. Secondary polynomial models were obtained using nonlinear regression analysis. The developed models were validated with mean square error, bias factor, and accuracy factor, which were 0.0003, 0.96, and 1.11, respectively, for specific growth rate and 7.69, 0.99, and 1.04, respectively, for lag time. The treatment of chlorine and UV did not change the color and texture of chicken breast after 7 d of storage at 4°C. As a result, the combination of chlorine at 100 mg/kg and UV at 300 mW·s/cm(2) appears to an effective method into inhibit L. monocytogenes growth in broiler carcasses with no negative effects on color and textural quality. Based on the validation results, the predictive models can be used to accurately predict L. monocytogenes growth in chicken breast.


Subject(s)
Chlorine/pharmacology , Food Microbiology , Food Preservation/methods , Listeria monocytogenes/drug effects , Listeria monocytogenes/radiation effects , Meat/microbiology , Ultraviolet Rays , Animals , Chickens
9.
Poult Sci ; 93(1): 208-15, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24570441

ABSTRACT

This study was conducted to evaluate the efficacy of sodium hypochlorite (NaOCl, 0-200 mg/kg), thiamine dilauryl sulfate (TDS, 1,000 mg/kg), and ultrasound (37 kHz, 380 W) on reducing Salmonella Typhimurim, mesophilic aerobic bacteria (MAB), and coliforms on chicken skin. Chemical and physical treatments were applied for 5 min either singly or jointly, and Salmonella previously inoculated on chicken skin were quantitatively assessed using brilliant green agar, and the populations of MAB and coliforms in the native flora were enumerated using plate count agar and violet red bile agar, respectively. In the evaluation of bacterial attachment/detachment, chicken skin was quantitatively assessed for loosely, intermediately, and tightly attached bacteria. The treatment effects on bacteria detachment were also visualized using field emission scanning electron microscopy. In addition, color and textural properties of the skin after treatments were evaluated using a color difference meter and texture analyzer. Antimicrobial activity of NaOCl increased as the NaOCl concentration was increased, especially for loosely attached cells. The combination of 200 mg/kg NaOCl and ultrasound (NaOCl/ultrasound) significant reduced loosely, intermediately, and tightly attached bacteria populations by 0.75 to 0.47, 0.43 to 0.41, and 0.83 to 0.54 log cfu/g for MAB, coliforms, and Salmonella Typhimurium, respectively. However, the combination of NaOCl and TDS (NaOCl/TDS) did not sufficiently reduce those cells on chicken skins, except for loosely attached MAB and coliforms. The NaOCl/ultrasound combination produced a higher reduction in numbers of inoculated and native bacteria flora than any single application, with no negative effect on skin color or texture. Generally, the loosely attached bacteria were less resistant to the chemical and physical treatments than the intermediately and tightly attached bacteria in chicken skin, presumably due to their location in deeper skin layer and crevices. Further research is needed to investigate how the intermediately and tightly attached microorganisms can be effectively eliminated from chicken skin.


Subject(s)
Chickens , Salmonella typhimurium/drug effects , Skin/microbiology , Animals , Disinfectants , Food Microbiology
10.
Article in English | MEDLINE | ID: mdl-23822106

ABSTRACT

This study was conducted to help better understand the current sodium intake of Korean children and to establish children's good eating habits through investigation of the sodium content of ready-to-eat foods collected from nine major amusement parks in Korea. The sodium content of a total of 322 products was analysed by using ICP and then the potential risk based on the recommended daily intake of sodium as described in the Korean dietary reference intakes was determined. The results showed that sodium content was the lowest in muffins (245 mg/100 g) and the highest in seasoned dried filefish (1825 mg/100 g). The average amounts of sodium per serving of seasoned dried filefish, tteokbokki and fish paste were 1150, 1248 and 1097 mg, respectively. The values were above 50% of the daily intake of sodium recommended by the Korean dietary reference intake. The ready-to-eat foods were also classified into high, medium and low sodium content on the basis of standards recommended by the Korean Food and Drug Administration. Most snacks were classified as high sodium foods because they exceeded "300 mg (84.5% of the total daily allowance)". Furthermore, the meal substitution foods such as kimbab, tteokbokki, mandus, sandwiches and hamburgers exceeded "600 mg (90.3% of the total daily allowance)" and were also classified as high sodium foods. In addition, ready-to-eat foods in amusement parks are similar to foods eaten on streets and foods around school zones, which contain high sodium content; thus, the intake frequency might be high, which would induce high risk to children health. Koreans already consume a high amount of sodium daily via their usual diets. So, the sodium content in snacks and substitution foods needs to be reduced. Consequently, this study noted that parents and guardians should carefully consider their children's consumption of ready-to-eat foods from Korean amusement parks.


Subject(s)
Diet/adverse effects , Fast Foods/analysis , Food Services , Leisure Activities , Sodium, Dietary/analysis , Child , Child, Preschool , Databases, Factual , Diet/ethnology , Electrochemical Techniques , Fast Foods/adverse effects , Fast Foods/economics , Fast Foods/standards , Female , Fish Products/adverse effects , Fish Products/analysis , Fish Products/economics , Fish Products/standards , Food Services/economics , Guideline Adherence , Health Promotion , Humans , Leisure Activities/economics , Male , Meals/ethnology , Recommended Dietary Allowances , Republic of Korea , Risk Assessment , Snacks/ethnology , Sodium, Dietary/adverse effects
11.
Article in English | MEDLINE | ID: mdl-23521141

ABSTRACT

The dietary intakes of nine synthetic food colours--amaranth, erythrosine, Allura Red, Ponceau 4R, tartrazine, Sunset Yellow FCF, Fast Green FCF, Brilliant Blue FCF and indigo carmine--permitted in Korea were estimated based on food consumption data for consumers and their concentrations in processed foods. The estimated daily intakes (EDIs) by Korean consumers were compared with the acceptable daily intakes (ADIs) of the colours. Among 704 foods sampled, 471 contained synthetic colours. The most highly consumed synthetic colours were Allura Red and tartrazine; the highest EDI/ADI ratios were found for amaranth, erythrosine and Allura Red. The EDIs of infants and children were higher than those of adults. The main food categories containing colours were beverages and liquor for adults, and beverages, chocolate and ice cream for infants and children. For average Korean consumers, the EDIs were not greater than 2.5% of their corresponding ADIs, although the EDI of a conservative consumer in the upper 95th percentile reached 37% of the ADI.


Subject(s)
Environmental Exposure , Food Coloring Agents , Child , Child, Preschool , Chromatography, High Pressure Liquid , Female , Humans , Infant , Male , Republic of Korea , Spectrophotometry, Ultraviolet
12.
Article in English | MEDLINE | ID: mdl-19680927

ABSTRACT

The molecular structures of enniatins H, I, and MK1688 and beauvericin were investigated by liquid chromatography-tandem mass spectrometry (LC-MS/MS). MS fragmentation occurred by loss of -CO after opening of the cyclic molecule to carbonyl carbon, and cleavage of the peptide and ester bonds in the molecular structure. Fusarium oxysporum KFCC 11363P was tested for its ability to produce beauvericin and enniatins H, I, and MK1688 on five cereal substrates: rice, barley, maize, wheat, and Indian millet kernels. Furthermore, optimal conditions for the production of the four mycotoxins by the Fusarium isolate were examined on maize at four temperatures (15, 20, 25, and 30 degrees C) and at three moisture contents (10, 20, and 40%). Large amounts of beauvericin and enniatin H were present in maize cultures at 25 degrees C (232.4 and 196.4 microg g(-1), respectively). Enniatins I and MK1688 were maximally formed at 20 degrees C (221.5 and 180.2 microg g(-1), respectively). The optimal moisture contents for the production of enniatins H (196.4 microg g(-1)) and MK1688 (165.6 microg g(-1)), were 40%.


Subject(s)
Chromatography, High Pressure Liquid/methods , Depsipeptides/chemistry , Fusarium/metabolism , Tandem Mass Spectrometry/methods , Depsipeptides/biosynthesis , Molecular Structure , Temperature
13.
Neurosci Lett ; 302(1): 58-62, 2001 Apr 13.
Article in English | MEDLINE | ID: mdl-11278111

ABSTRACT

Although estrogen is known to exert beneficial effects on Alzheimer's disease, its underlying cellular mechanisms have not been clear. In this study we investigated whether or not neuroprotective effects of estrogen are mediated by estrogen receptors (ERs). Treatment of estrogen (1.8 nM) reduced beta-amyloid (Abeta)-induced death of ER-expressing W4 cells. This effect of estrogen was blocked by a specific ER blocker ICI 182,780. When estrogen was treated to HT22 cells, which lack functional ERs, Abeta-induced cell death was not affected. Transfection of HT22 cells with human ERalpha, but not ERbeta, restored protective action of estrogen against Abeta. Hoechst staining revealed that estrogen protected ERalpha-expressing cells by blocking Abeta-induced apoptosis. These results indicate that estrogen blocks Abeta-induced cell death via ERalpha-dependent pathways.


Subject(s)
Amyloid beta-Peptides/antagonists & inhibitors , Apoptosis/drug effects , Estradiol/analogs & derivatives , Estrogens/pharmacology , Neurons/drug effects , Receptors, Estrogen/drug effects , Amyloid beta-Peptides/pharmacology , Apoptosis/physiology , Cell Death/drug effects , Cell Death/physiology , Cells, Cultured , Estradiol/pharmacology , Estrogen Antagonists/pharmacology , Estrogen Receptor alpha , Estrogen Receptor beta , Fulvestrant , Humans , Neurons/metabolism , Receptors, Estrogen/metabolism , Transfection
14.
Glycoconj J ; 18(10): 779-87, 2001 Oct.
Article in English | MEDLINE | ID: mdl-12441667

ABSTRACT

The rfaE (WaaE) gene of Salmonella typhimurium is known to be located at 76min on the genetic map outside of the rfa gene cluster encoding core oligosaccharide biosynthesis of lipopolysaccharide(LPS). The rfaE mutant synthesizes heptose-deficient LPS; its LPS consists of only lipid A and 3-deoxy-D-manno-octulosonic acid (KDO), and the rfaE gene is believed to be involved in the formation of ADP-L-glycero-D-manno-heptose. Mutants, which make incomplete LPS, are known as rough mutants. Salmonella typhimurium deep-rough mutants affected in the heptose region of the inner core often show reduced growth rate, sensitivity to high temperature and hypersensitivity to hydrophobic antibiotics. We have cloned the rfaE gene of S. typhimurium. The chromosomal region carrying this gene was isolated by screening a genomic library of S. typhimurium using the complementation of S. typhimurium rfaE mutant. The 2.6-Kb insert in the plasmid pHEPs appears to carry a functional rfaE gene. SL1102 (rfaE543) makes heptose-deficient LPS and has a deep rough phenotype, but pHEPs complement the rfaE543 mutation to give the smooth phenotype. The sensitivity of SL1102 to bacteriophages (P22.c2, Felix-O, Br60) which use LPS as their receptor for adsorption is changed to that of wild-type strain. The permeability barrier of SL1102 to hydrophobic antibiotics (novobiocin) is restored to that of wild-type. LPS produced by SL1102 (rfaE543) carrying pHEPs makes LPS indistinguishable from that of smooth strains. The rfaE gene encoded a polypeptide of 477 amino acid residues highly homologous to the S. enterica rfaE protein (98% identity), E. coli (93% identity), Yersenia pestis (85% identity), Haemophilus influenzae (70% identity) and Helicobacter pyroli (41% identity) with a molecular weight 53 kDa.


Subject(s)
Bacterial Proteins/genetics , Glycosyltransferases , Lipopolysaccharides/biosynthesis , Salmonella typhimurium/genetics , Salmonella typhimurium/metabolism , Amino Acid Sequence , Bacterial Proteins/biosynthesis , Bacteriophages/physiology , Base Sequence , Blotting, Southern/methods , Carbohydrate Sequence , Cloning, Molecular , Genetic Complementation Test , Haemophilus influenzae/genetics , Lipopolysaccharides/chemistry , Molecular Sequence Data , Novobiocin/pharmacology , Plasmids , Restriction Mapping , Sequence Homology, Amino Acid
15.
Mol Cells ; 10(1): 13-7, 2000 Feb 29.
Article in English | MEDLINE | ID: mdl-10774741

ABSTRACT

The insect diuretic hormone (DH) binds to their receptor in malpighian tubules, and stimulates water secretion and cAMP synthesis. Complementary DNA encoding a diuretic hormone receptor was cloned from the malpighian tubules of Bombyx mori. The cloned cDNA encodes a protein consisting of 391 amino acid residues with the seven transmembrane domains. The receptor protein is homologous with that of other insects, and is structurally related to G-protein coupled receptors such as corticotropin relating factor (CRF), secretin, and vasoactive intestinal peptide.


Subject(s)
Bombyx/genetics , DNA, Complementary/genetics , Insect Hormones/metabolism , Insect Proteins , Receptors, Cell Surface/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , DNA, Complementary/chemistry , Malpighian Tubules/metabolism , Molecular Sequence Data , Receptors, Cell Surface/metabolism , Sequence Alignment , Sequence Analysis, DNA , Sequence Homology, Amino Acid
16.
Peptides ; 20(5): 561-8, 1999.
Article in English | MEDLINE | ID: mdl-10465507

ABSTRACT

A peptide with paralytic activity in larvae of the silkworm, Bombyx mori, was isolated from its hemolymph. Purification procedures consisted of extraction with 50% acetone, Vydac C4 reversed-phase cartridge elution and 4 steps of reversed-phase HPLC. Injection of the purified peptide into 4th instar B. mori larvae caused rapid and rigid paralysis for 2 min at a dose of 3.4 ng/larva. This paralytic peptide consists of 23 amino acid residues containing 2 cysteines with an intra-disulfide bond. The complete amino acid sequence is: H-Glu-AsnPhe-Val-Gly-Gly-Cys-Ala-Thr-Gly-Phe-Lys-Arg-Thr-Ala-Asp-G ly-Arg-Cys-Lys-Pro-Thr-Phe-OH. The relationship between structure and the biologic activity of synthetic analogs indicated that the entire amino acid sequence and the intra-disulfide bond were necessary for biological activity.


Subject(s)
Bombyx/chemistry , Hemolymph/chemistry , Neuropeptides/pharmacology , Paralysis/chemically induced , Amino Acid Sequence , Animals , Biological Assay , Disulfides/chemistry , Dose-Response Relationship, Drug , Larva/chemistry , Molecular Sequence Data , Neuropeptides/chemistry , Sequence Analysis , Structure-Activity Relationship
17.
J Nutr ; 127(3): 449-57, 1997 Mar.
Article in English | MEDLINE | ID: mdl-9082029

ABSTRACT

An analysis of viable bacterial populations enumerated on carbohydrate selective media was used to simulate the colonic environment in vitro and determine if differential media could detect significant microbial shifts due to dietary fiber source, dietary fat source, and carcinogen. Male Sprague-Dawley rats were provided with either pectin or cellulose as a fiber source, either corn or fish oil as a source of fatty acids, and injected with either azoxymethane (AOM), a gastrointestinal carcinogen, or saline in a 2 x 2 x 2 factorial design. At 6 and 10 mo of age, fresh feces were collected, homogenized in anaerobic buffer and anaerobically plated onto differential media. Diets containing pectin supported more anaerobes at 6 mo of age (P < 0.01) than diets containing cellulose. Rats injected with AOM and consuming either pectin or corn oil supported more anaerobes at 10 mo of age (P < 0.05) than rats injected with saline and consuming the same diets. Rats consuming cellulose and receiving AOM but not expressing tumors possessed larger anaerobic populations at 10 mo of age (P < 0.05) than rats consuming cellulose, injected with AOM and expressing tumors. These effects show that gastrointestinal bacterial populations, as measured by carbohydrate specific media, respond to dietary changes such as dietary fiber source, and thus may play a key role in the etiology of colon cancer.


Subject(s)
Bacteria/growth & development , Carcinogens/pharmacology , Colon/microbiology , Dietary Fats/pharmacology , Dietary Fiber/pharmacology , Feces/microbiology , Aging/metabolism , Animals , Anti-Infective Agents, Local , Azoxymethane/pharmacology , Bacteria/drug effects , Cellulose/pharmacology , Cetrimonium , Cetrimonium Compounds , Colonic Neoplasms/chemically induced , Colonic Neoplasms/microbiology , Colony Count, Microbial , Coloring Agents , Congo Red , Corn Oil/pharmacology , Culture Media , Fish Oils/pharmacology , Male , Pectins/pharmacology , Rats , Rats, Sprague-Dawley
18.
Adv Exp Med Biol ; 412: 201-8, 1997.
Article in English | MEDLINE | ID: mdl-9192013

ABSTRACT

Salmonella typhimurium is a significant hazard to consumer health that is carried asymptomatically in poultry gastrointestinal tracts. Nurmi cultures may prevent Salmonella colonization in young chicks, but the mechanism of competitive exclusion is unclear. Modeling Salmonella's metabolism in pure culture may allow for greater definition in choosing strains for Nurmi cultures. The growth rates and affinity constants of S. typhimurium growing in amino acid-limited conditions were determined in batch culture and compared to primary poultry isolates of cecal strains. Serine and NH4Cl were the best N sources for growth of all organisms tested in this study. The fermentation response of S. typhimurium was also monitored in continuous culture at a slow dilution rate of 0.021 h-1. S. typhimurium was found to adapt to VL media, with trends in protein disappearance, Yglucose, and Yprotein. This may show that amino acid or protein concentrations may be an integral component of the initial establishment of S. typhimurium in the cecum.


Subject(s)
Salmonella typhimurium/growth & development , Amino Acids/metabolism , Anaerobiosis , Animals , Cecum/microbiology , Chickens/microbiology , Escherichia coli/growth & development , Fermentation
19.
J Food Prot ; 58(12): 1335-1339, 1995 Dec.
Article in English | MEDLINE | ID: mdl-31159035

ABSTRACT

The objective of this in vitro experimentation was to compare the survivability of Salmonella typhimurium strains and selected facultative chicken cecal bacteria after specific amino acid-limited growth on either serine, threonine, arginine, or aspartate. Survivability of Salmonella typhimurium and chicken cecal bacteria was estimated by measuring the rate of decrease of viable cell numbers and calculating the average time for 50% of the cells to become nonviable (50% survival time, ST50). Two S. typhimurium strains, LT2 and a primary poultry isolate (NO/NA), and three selected facultative chicken cecal bacteria, Citrobacter freundii , Escherichia coli and Escherichia fergusonii , were grown aerobically at 37°C to stationary phase on carbon-limited or nitrogen-limited minimal media. All organisms remained viable longer (P < 0.05) on serine media than on any of the other media tested. When serine was used as a nitrogen source in minimal media the ST50 of C. freundii and E. fergusonii were significantly longer than those of the two S. typhimurium strains. It appears that when media are limited in the same nutrient, the ability to sustain viability varies among facultative bacteria derived from the chicken cecum.

20.
J Food Prot ; 57(12): 1074-1079, 1994 Dec.
Article in English | MEDLINE | ID: mdl-31121649

ABSTRACT

Our objective in this study was to use batch culture to estimate growth kinetic parameters for a growth-limiting amino acid, serine, in Salmonella typhimurium and a in chicken cecal bacterium, Escherichia fergusonii , and to test these predictions for competitiveness in a mixed culture of the two organisms. Under anaerobic growth conditions, the two bacteria grew only when serine was provided as the nitrogen source, When serine was used as a carbon source in aerobic media, the maximum growth rates of the two organisms were considerably lower and the affinity constants were higher than when serine was used as a nitrogen source. The maximum growth rates of the two organisms were lower in anaerobic media than in aerobic media, but serine anaerobic affinity constants were lower than those from aerobic media. In aerobic mixed culture, S. typhimurium outgrew E. fergusonii in N-limited minimal media containing 0.1 mM serine but in N-nonlimited minimal media containing 10 mM serine, E. fergusonii outgrew S. typhimurium . In anaerobic mixed culture, E. fergusonii outgrew S. typhimurium in media containing both 0.1 mM and 10 mM serine concentrations. It appears that the cause of inhibition of Salmonella with serine as the variable nutrient differs depending on the oxidation-reduction status.

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