Biomedical subjects

D L Marshall

Publications and source records attributed to D L Marshall.

At least 19 recordsLinked to original sources

PubMed2002

Growth and survival of Vibrio parahaemolyticus in postharvest American oysters.

Oysters at the retail stage of distribution generally contain greater densities of Vibrio parahaemolyticus than do oysters at harvest. The objective of this study was to determine the effects of postharvest storage at 26 and 3 degrees C on the growth and survival of naturally occurring V. parahaemolyticus in shellstock American oysters (Crassostrea virginica). Oysters were collected monthly from May 1998 through April 1999 from Mobile Bay, Alabama, and their V. parahaemolyticus densities were determined after 0, 5, 10, and 24 h of postharvest storage at 26 degrees C. After 24 h of storage at 26 degrees C, oysters were transferred to a refrigerator at 3 degrees C and analyzed 14 to 17 days later. V. parahaemolyticus numbers were determined by a direct plating method involving an alkaline-phosphatase-labeled DNA probe that targets the species-specific thermolabile hemolysin gene (tlh-AP) to identify suspect isolates. From April to December, when water temperatures at harvest were >20 degrees C, the geometric mean harvest density of V. parahaemolyticus was 130 CFU/g. When water temperatures were <20 degrees C, the geometric mean harvest density was 15 CFU/g. After harvest, V. parahaemolyticus multiplied rapidly in live oysters held at 26 degrees C, showing a 50-fold increase (1.7 log CFU/g) at 10 h and a 790-fold increase (2.9 log CFU/g) at 24 h (April through December). Average V. parahaemolyticus numbers showed a sixfold decrease (0.8 log CFU/g) after approximately 14 days of refrigeration. These results indicate that V. parahaemolyticus can grow rapidly in unrefrigerated oysters.

Animals↗

PubMed2001

Evaluation of two direct plating methods using nonradioactive probes for enumeration of Vibrio parahaemolyticus in oysters.

Oysters (Crassostrea virginica) were collected monthly from May 1998 to April 1999 from Mobile Bay, Ala., and analyzed to determine Vibrio parahaemolyticus densities at zero time and after 5, 10, and 24 h of postharvest storage at 26 degrees C. After 24 h of storage at 26 degrees C, oysters were transferred to a refrigerator at 3 degrees C and then analyzed 14 to 17 days later. The V. parahaemolyticus numbers were determined by the most-probable-number procedure using alkaline phosphatase-labeled DNA probe VPAP, which targets the species-specific thermolabile hemolysin gene (tlh), to identify suspect isolates (MPN-VPAP procedure). Two direct plating methods, one using a VPAP probe (Direct-VPAP) and one using a digoxigenin-labeled probe (Direct-VPDig) to identify suspect colonies, were compared to the MPN-VPAP procedure. The results of the Direct-VPAP and Direct-VPDig techniques were highly correlated (r = 0.91), as were the results of the Direct-VPAP and MPN-VPAP procedures (r = 0.91). The correlation between the Direct-VPDig and MPN-VPAP results was 0.85. The two direct plating methods in which nonradioactive DNA probes were used were equivalent to the MPN-VPAP procedure for identification of total V. parahaemolyticus, and they were more rapid and less labor-intensive.

Alkaline Phosphatase↗

PubMed2001

Antacid increases survival of Vibrio vulnificus and Vibrio vulnificus phage in a gastrointestinal model.

Viable counts of three strains of Vibrio vulnificus and its phage were determined during exposure to a mechanical gastrointestinal model with or without antacid for 9 h at 37 degrees C. V. vulnificus was eliminated (>4-log reduction) within 30 min in the gastric compartment (pH decline from 5.0 to 3.5). Viable V. vulnificus cells delivered from the gastric compartment during the first 30 min of exposure reached 10(6) to 10(8) CFU/ml in the intestinal compartment after 9 h (pH 7.0). Phages were eliminated within 45 min in the gastric compartment (pH decline from 5.1 to 2.5). Less than a 2-log reduction of phage was observed in the intestinal compartment after 9 h (pH 7.0). When the gastric compartment contained antacid V. vulnificus counts decreased slightly (<2 log) during 2 h of exposure (pH decline from 7.7 to 6.0), while counts in the intestinal compartment (pH 7.5) reached 10(7) to 10(9) CFU/ml. Phage numbers decreased 1 log after 2 h in the gastric compartment (pH decline from 7.7 to 5.7) containing antacid and decreased 1 log in the intestinal compartment (pH 7.6) after 9 h. Presence of antacid in the gastric compartment of the model greatly increased the ability of both V. vulnificus and its phage to survive simulated gastrointestinal transit and may be a factor involved with oyster-associated illness.

Animals↗

PubMed2001

Mechanisms of differential pollen donor performance in wild radish, Raphanus sativus (Brassicaceae).

In order to understand the characters on which sexual selection might operate in plants, it is critical to assess the mechanisms by which pollen competition and mate choice occur. To address this issue we measured a number of postpollination characters, ranging from pollen germination and pollen tube growth to final seed paternity, in wild radish. Crosses were performed using four pollen donors on a total of 16 maternal plants (four each from four families). Maternal plants were grown under two watering treatments to evaluate the effects of maternal tissue on the process of mating. The four pollen donors differed significantly in number of seeds sired and differed overall in the mating characters measured. However, it was difficult to associate particular mechanistic characters with ability to sire seeds, perhaps because of interactions among pollen donors within styles or among pollen donors and maternal plants. The process of pollen tube growth and fertilization differed substantially among maternal watering treatments, with many early events occurring more quickly in stressed plants. Seed paternity, however, was somewhat more even among pollen donors used on stressed maternal plants, suggesting that when maternal tissue is more competent, mating is slowed and is more selective.

Journal Article↗

PubMed2000

Effect of simulated gastric fluid and bile on survival of Vibrio vulnificus and Vibrio vulnificus phage.

Bacteria and phages may be exposed to acid conditions in the stomach and to bile in the intestine. Survival of three strains of Vibrio vulnificus and three strains of its phages was examined at 37 degrees C after exposure to simulated gastric fluid at pH 3 to 4 or to 0, 1, and 2% bile in broth or buffer. Mean D-values (decimal reduction times) at pH 4 and 3 were 3.3 and 1.3 min for V. vulnificus and 97.8 and 0.7 min for its phages. No V. vulnificus survivors were found at pH 2.0. There were few survival differences among strains of V. vulnificus or its phages. Numbers of V. vulnificus increased 1 log in tryptic soy broth containing 1 or 2% bile after 3 h. Numbers of V. vulnificus and its phages remained constant in phosphate-buffered saline regardless of bile concentrations up to 3 h. Those V. vulnificus bacteria and phages that survive stomach acidity may proliferate in the small intestine, since they are resistant to bile.

Bacteriophages↗

PubMed2000

Impact of acid on survival of Vibrio vulnificus and Vibrio vulnificus phage.

Three strains of Vibrio vulnificus and V. vulnificus phages were tested for acid sensitivity at 21 degrees C. V. vulnificus strain 304 was more resistant to pH 4.0 than strains CVD-1 and A-9, whereas acid sensitivities of V. vulnificus strains at pH 3.0 and 2.0 were similar. V. vulnificus phage strain 110A-7 was more resistant to pH 4.0 than strain 153A-7, whereas acid sensitivities of phage strains at pH 3.5 and 3.0 were similar. Numbers of V. vulnificus and its phage were close to the limit of detection after 100 s at pH 2.0 and after 24 min at pH 3.0. Acid D-values at 21 degrees C decreased as pH decreased for both V. vulnificus and phages. D-values of phage strains at pH 3.5 were 10-fold greater than those of host strain at pH 4.0. D-values of phage strains were slightly greater than those of host strain at pH 3.0. These results suggest that V. vulnificus and its phage were very sensitive to pH of less than 3.0, although V. vulnificus phages were more resistant to acid than their host.

Animals↗

PubMed2000

Effects of pollen load size and composition on pollen donor performance in wild radish, Raphanus sativus (Brassicaceae).

A critical concern in the debate over the importance of sexual selection in plants is whether the nonrandom mating demonstrable in greenhouse crosses can occur in the field. Field populations likely experience smaller and more variable pollen load sizes than those that have been used in many greenhouse experiments. Therefore, we performed a greenhouse experiment in which we varied both pollen load size and composition in wild radish, Raphanus sativus, and examined the paternity of seeds. We used five maternal plants and four pairs of pollen donors. We were able to produce pollen loads of 40, 118, and 258 grains per stigma. The smallest of the pollen loads was scant enough to result in a slight, but significant reduction in seed number per fruit. While variation in pollen load composition significantly affected the proportions of seeds fathered by different donors, variation in pollen load size did not. The relative performance of different donors was constant across pollen load sizes, suggesting that, for this species, differential performance of pollen donors can occur at pollen load sizes that are likely to occur in field populations.

Journal Article↗

PubMed1998

Organic acid dipping of catfish fillets: effect on color, microbial load, and Listeria monocytogenes.

Microbiological and color changes of catfish fillets were determined following dip treatment in solutions at 4 degrees C of 2% acetic, citric, hydrochloric, lactic, malic, or tartaric acid. Fillets were inoculated with an eight-strain mixture of Listeria monocytogenes prior to dipping. L. monocytogenes, coliform, and aerobic plate counts and surface pH and Hunter color were measured at 0, 2, 5, and 8 days of storage at 4 degrees C. Acid dipping reduced surface pH and L. monocytogenes, coliform, and aerobic microbial loads. Little microbial proliferation was observed on acid-treated fillets, however, controls had a distinct foul odor and microbial loads in excess of 10(6) CFU/g by day 8. On untreated fillets, L. monocytogenes counts did not increase during storage, perhaps due to competitive inhibition by normal catfish microflora. Hunter color analysis revealed lighter and yellower acid-treated fillets than untreated controls, with malic acid producing the least bleaching. The shelf life of refrigerated fillets increased when fillets were acid dipped. It remains to be established if this enhanced microbial quality also parallels sensory acceptability.

Animals↗

PubMed1997

Excitability of neural elements within the rat corpus striatum.

The excitability of cholinergic, glutamatergic and dopaminergic elements within the rat neostriatum was studied in both in vivo and in vitro preparations. In vivo, the microdialysis technique was used to measure the release of striatal acetylcholine and dopamine under basal and electrically evoked conditions. For comparison, acetylcholine, dopamine and glutamate release was assayed in media obtained from superfused rat striatal slices. Electrical stimulation was used to derive the strength-duration functions and their chronaxies of stimulated elements containing the three neurotransmitter types. The chonaxies for experiments in vitro and in vivo were similar: the chronaxy values for elements containing acetylcholine were the shortest, the values for glutamate were intermediate, and the values for those containing dopamine were the longest. Based on the chronaxy estimates, it is proposed that the elements containing acetylcholine are the large cholinergic interneurons of striatum, and the elements containing glutamate and dopamine are the terminals of corticostriatal and nigrostriatal neurons, respectively. These results indicate that electrical stimulation of neural elements surrounding a microdialysis probe can be an additional tool to examine the factors that regulate neurotransmitter release. Likewise, investigators can activate specific striatal elements by using pulse durations that coincide with their chronaxies.

Acetylcholine↗

PubMed1997

Presynaptic nicotinic modulation of dopamine release in the three ascending pathways studied by in vivo microdialysis: comparison of naive and chronic nicotine-treated rats.

The modulation of dopamine release by presynaptic nicotinic receptors in vitro is well established, but the significance of this effect in vivo is unclear. We have characterised the effect of nicotine, locally applied via a microdialysis probe, on dopamine release from the terminal regions of three ascending dopaminergic pathways in conscious, freely moving rats. Nicotine caused a dose-dependent increase in dopamine release in the striatum, the nucleus accumbens, and, to a lesser extent, the frontal cortex. Metabolite levels were unaltered by any concentration of nicotine. Prior administration of mecamylamine via the probe abolished the nicotine-evoked increase in dopamine release, confirming the mediation of nicotinic receptors. The dose dependence of mecamylamine-sensitive, nicotine-evoked dopamine release was similar in all three brain regions. However, 10(-5) M tetrodotoxin totally blocked nicotine-stimulated dopamine release in the striatum and the accumbens but not the cortex. Daily subcutaneous injections of nicotine (0.4 mg kg-1 for 7 days) increased the response to a subsequent local application of nicotine in the striatum, and a similar trend was found in the other brain areas. The same daily dose of nicotine given as a continuous infusion had no effect, whereas infusion of 4 mg kg-1 day-1 increased the response to a subsequent nicotine challenge. The localisation and regulation of nicotinic receptors in the terminal fields of dopaminergic pathways are discussed.

Animals↗

PubMed1996

Effect of monolaurin and lactic acid on Listeria monocytogenes attached to catfish fillets.

The purpose of this study was to determine the effects of monolaurin and lactic acid, singly or combined, on Listeria monocytogenes attached to catfish fillets. Skinless catfish fillets were inoculated with L. monocytogenes and dip treated in monolaurin and/or lactic acid solution for various time periods. Results showed that monolaurin up to 400 micrograms/ml had no influence on counts. Conversely, lactic acid-treated fillets had reduced counts compared to controls. Dipping in 0.85, 1.70, or 2.55% lactic acid for 30 min reduced counts by 0.9, 1.4, or 1.3 logs, respectively. Extending the dipping time to 60 min resulted in little additional decrease in counts. Combining monolaurin with lactic acid yielded results similar to lactic acid alone. Hence, population reduction ability resides with lactic acid and not monolaurin.

Animals↗

PubMed1996

Antimicrobial susceptibility and plasmid-mediated streptomycin resistance of Plesiomonas shigelloides isolated from blue crab.

Five Plesiomonas shigelloides strains isolated from retail or wild-catch Louisiana blue crabs (Callinectes sapidus) were examined for resistance to selected antibiotics and presence of plasmids. Three agar media, MacConkey, Salmonella-Shigella and a modification of inositol brilliant green bile salt (IBB), were evaluated for isolation of P. shigelloides. Of these, IBB was most satisfactory for distinguishing between P. shigelloides and the related species Aeromonas hydrophila. Isolates were susceptible to gentamicin, nalidixic acid and tetracycline, and were resistant to ampicillin, carbenicillin, kanamycin and streptomycin. Each isolate carried three plasmids of approximately 2.5 kb, 3.8 kb and 5.3 kb. Plasmid curing with ethidium bromide and elevated temperature linked the streptomycin resistance determinant with the 3.8 kb and/or 5.3 kb plasmids. These results provide the first description of small size plasmids in P. shigelloides. Identical antibiotic resistance patterns of the isolates indicated they share a common environment and likely share a common mode for developing antibiotic resistance. Furthermore, emergence of antibiotic-resistant P. shigelloides may imply contamination of Louisiana aquatic environments by wastewater.

Animals↗

PubMed1996

Monolaurin and acetic acid inactivation of Listeria monocytogenes attached to stainless steel.

Individual and combined antimicrobial effects of monolaurin and acetic acid on Listeria monocytogenes planktonic cells or stainless-steel-adherent cells were determined in order to evaluate cell viability during a 25-min exposure period at 25 degrees C. A 10(7)-colony-forming units (CFU)/ml population of planktonic cells was completely inactivated by the synergistic combination of 1% acetic acid with 50 or 100 microg/ml of monolaurin within 25 or 20 min, respectively. Either compound alone caused partial but incomplete inactivation within the same time periods. A population of 10(5) CFU/cm2 of 1-day adherent cells on stainless steel was completely inactivated within 25 min, but with the highest concentrations of the combined chemicals, i.e., 1% acetic acid and 100 microg/ml of monolaurin. The combined chemical treatment again synergistically produced greater inhibition. A 10(6)-CFU/cm2 population of 7-day adherent cells was not completely inactivated within 25 min of exposure, although counts did decline. The results demonstrate increased resistance of attached L. monocytogenes to acetic acid and monolaurin and show that resistance increased with culture age. Combinations of organic acids and monolaurin might be considered as sanitizers of food contact surfaces, but activities of such combinations are likely to be less than other commonly used sanitizers.

Acetic Acid↗

PubMed1996

Combined effects of packaging atmosphere and lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat 4 degrees C.

The effect of lactic acid on growth and survival of Listeria monocytogenes in crayfish tail meat stored under refrigeration and various gas environments was investigated. Frozen crayfish tail meat was thawed overnight, autoclaved, cooled, and inoculated with approximately 4 log colony-forming units (CFU) of a mixed-strain (Scott A and F5027) L. monocytogenes culture per gram of meat. Inoculated samples were blended with 0, 0.5, 1.0, 1.5, or 2.0% lactic acid and packaged under air, vacuum, or modified atmosphere (74.8% CO2, 10.4% O2, and 14.8% N2) and stored at 4 degrees C for 20 days. Results demonstrated that modified atmosphere packaging inhibited the growth of L. monocytogenes more than air and vacuum packaging at 0 and 1% lactic acid. Microbial counts declined steadily in crayfish tail meat treated with 2% lactic acid, with no differences among the packaging atmospheres. The lag phase was extended by 8 days in samples treated with 1% lactic acid and modified atmosphere compared to that in air or vacuum packaging. Overall, the combination of lactic acid and modified atmosphere had the greatest potential to prevent growth of L. monocytogeines.

Animals↗

PubMed1994

Neuromuscular effects of some potassium channel blocking toxins from the venom of the scorpion Leiurus quinquestriatus hebreus.

The scorpion venom Leiurus quinquestriatus hebreus was fractionated by chromatography in order to isolate toxins that affected binding of radiolabelled dendrotoxin to K+ channel proteins on synaptosomal membranes and that facilitated acetylcholine release in chick biventer cervicis nerve-muscle preparations. In addition to the previously characterized charybdotoxin, three toxins were isolated: 14-2, 15-1 and 18-2. Toxin 14-2 has a blocked N-terminus and because of low quantities, it has not been sequenced; 15-1 is a newly sequenced toxin of 36 residues with some overall homology to charybdotoxin and noxiustoxin; 18-2 is identical to charybdotoxin-2. The apparent Ki against dendrotoxin binding were: charybdotoxin, 3.8 nM; 14-2, 150 nM; 15-1, 50 nM; and 18-2, 0.25 nM. Toxin 14-2 (75 nM-1.5 microM) had a presynaptic facilitatory effect on neuromuscular preparations. Toxin 15-1 augmented responses to direct muscle stimulation, probably because it blocked Ca(2+)-activated K+ currents in muscle fibres. Toxin 18-2 (charybdotoxin-2) had a potent presynaptic facilitatory action, with less effect on direct muscle stimulation. This contrasts with the relatively weak neuromuscular effects of the highly homologous charybdotoxin. On a Ca(2+)-activated K+ current in mouse motor nerve endings, charybdotoxin and toxin 18-2 produced maximal block at around 100 nM, whereas 15-1 was inactive at 300 nM. Charybdotoxin can increase quantal content, but this is more likely to result from block of voltage-dependent K+ channels than Ca(2+)-activated channels: the increase in transmitter release occurred in conditions in which little IKCa would be present; higher concentration of charybdotoxin and longer exposure times were required to increase transmitter release than those needed to block IKCa, and the facilitatory effects of charybdotoxin and toxin 18-2 correlated more with their effects on dendrotoxin binding than on block of IKCa.

Acetylcholine↗

PubMed1994

Mechanism of inhibited growth of Bacillus pumilus by Propionibacterium freudenreichii subsp. shermanii.

Physiological studies were conducted in an attempt to elucidate the mechanism of inhibition of Bacillus pumilus by Propionibacterium freudenreichii subsp. shermanii. Inhibition of B. pumilus by P. shermanii occurred in media supplemented with 1% glucose, indicating that glucose utilization by the latter bacterium was not responsible for growth inhibition of the former bacterium. The medium pH in which P. shermanii inhibited the growth of B. pumilus was 4.3. Propionic acid was positively identified in the culture medium in which B. pumilus was inhibited by P. shermanii. The presence of propionic acid and a low medium pH may account for the inhibition of B. pumilus by P. shermanii. Sodium lactate concentrations of 0.8-1.0% were essential for the continuous growth of and propionic acid production by P. shermanii. Thus, use of P. shermanii to inhibit B. pumilus in foods would likely require a lactate source.

Antibiosis↗

PubMed1993

Improved method for the routine analysis of acetylcholine release in vivo: quantitation in the presence and absence of esterase inhibitor.

An improved high-performance liquid chromatographic (HPLC) method using electrochemical detection (ED) is described capable of routinely measuring the low levels of acetylcholine (ACh) typically found in rat brain microdialysis samples. Microdialysis was performed in the striatum of the urethane anesthetized rat using a 4-mm membrane length, high recovery (40% at 1.0 microliters/min; ambient conditions), loop-design probe perfused with an artificial cerebrospinal fluid (aCSF) solution containing physiologically normal calcium levels (1.2 mM). The HPLC method utilizes a polymeric stationary phase to resolve choline (Ch) from ACh. These analytes are then converted to hydrogen peroxide (H2O2) by a solid-phase reactor (containing immobilized choline oxidase and acetylcholinesterase enzymes). The H2O2 is detected amperometrically and quantitated on a platinum (Pt) working electrode (+300 mV; with a unique analytical cell featuring a solid-state palladium reference electrode). Two designs of the Pt working electrode were examined, differing only in the support material used (Kel-F or PEEK). The Kel-F/Pt electrode had a limit of detection (LOD) for both analytes of < 30 fmol per 10 microliters with a signal-to-noise ratio of 3:1. Striatal microdialysis perfusates were monitored for ACh and Ch over a 0-1000 nM range of neostigmine (NEO) in the CSF perfusion medium. Using the 4-mm probe, basal ACh and Ch levels were detected with a NEO level as low as 10 nM and were found to be 37 +/- 3 fmol and 22 +/- 1 pmol per 10 microliters (mean +/- S.E.M., n = 6 replicates) respectively. In similar experiments using 3-mm concentric probes comparable (lower) levels of ACh were found with the 50 and 1000 nM NEO doses (n = 4-21 animals). ACh could not be reliably quantitated when animals were perfused with the 10 nM dose of NEO (n = 4). The PEEK/Pt electrode had an improved LOD of < 20 fmol per 10 microliters due to a two- to three-fold decrease in the background noise component. Basal striatal levels of ACh in the absence of NEO approached the LOD and were found to be 15 +/- 2 fmol per 10 microliters; Ch was 5 +/- 1 pmol per 10 microliters (n = 2, mean of five basal samples). The analytical system requires very little maintenance; a simple electrochemical electrode cleaning step eliminates the need for routine polishing of the Pt electrode and the mobile phase is stable for up to one week.(ABSTRACT TRUNCATED AT 400 WORDS)

Acetylcholine↗

PubMed1993

Effect of choline on basal and stimulated acetylcholine release: an in vivo microdialysis study using a low neostigmine concentration.

Using in vivo microdialysis, we examined the ability of choline (Ch) chloride (120 mg/kg i.p.) to amplify basal and stimulated acetylcholine (ACh) release from rat striatum in the presence of high (10(-5) M) and low (5 x 10(-8) M) neostigmine concentration. High concentrations might suppress ACh release, and thus Ch dependence, by excessively stimulating presynaptic cholinergic receptors; alternatively, they could enhance Ch dependence by depriving the cholinergic terminals of Ch that would otherwise be formed intrasynaptically from the hydrolysis of ACh. Both basal and stimulated ACh release were found to be tetrodotoxin (TTX) sensitive. The concentration of neostigmine in the microdialysis fluid positively affected basal ACh levels, but had no effect on Ch levels. Ch administration significantly increased ACh release (to 136% of basal values; P < 0.01) in the presence of the low neostigmine concentration, but failed to significantly increase ACh release following local electrical depolarization of striatal neurons. In contrast, Ch failed to affect basal ACh release in the presence of the high neostigmine concentration, but did increase electrically evoked release to 408% of basal values, as compared with 250% in rats receiving saline instead of the Ch (P < 0.05). Ch administration significantly increased microdialysate Ch levels in the presence of both of the neostigmine concentrations. Local administration of oxotremorine, a muscarinic agonist, to animals receiving the lower neostigmine concentration reduced basal ACh release and reduced the increase in basal release produced by Ch administration.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗