Dynamics of maternal and paternal effects on embryo and seed development in wild radish (Raphanus sativus).

BACKGROUND AND AIMS: Variability in embryo development can influence the rate of seed maturation and seed size, which may have an impact on offspring fitness. While it is expected that embryo development will be under maternal control, more controversial hypotheses suggest that the pollen donor and the embryo itself may influence development. These latter possibilities are, however, poorly studied. Characteristics of 10-d-old embryos and seeds of wild radish (Raphanus sativus) were examined to address: (a) the effects of maternal plant and pollen donor on development; (b) the effects of earlier reproductive events (pollen tube growth and fertilization) on embryos and seeds, and the influence of embryo size on mature seed mass; (c) the effect of water stress on embryos and seeds; (d) the effect of stress on correlations of embryo and seed characteristics with earlier and later reproductive events and stages; and (e) changes in maternal and paternal effects on embryo and seed characteristics during development. METHODS: Eight maternal plants (two each from four families) and four pollen donors were crossed and developing gynoecia were collected at 10 d post-pollination. Half of the maternal plants experienced water stress. Characteristics of embryos and seeds were summarized and also compared with earlier and later developmental stages. KEY RESULTS: In addition to the expected effects of the maternal plants, all embryo characters differed among pollen donors. Paternal effects varied over time, suggesting that there are windows of opportunity for pollen donors to influence embryo development. Water-stress treatment altered embryo characteristics; embryos were smaller and less developed. In addition, correlations of embryo characteristics with earlier and later stages changed dramatically with water stress. CONCLUSIONS: The expected maternal effects on embryo development were observed, but there was also evidence for an early paternal role. The relative effects of these controls may change over time. Thus, there may be times in development when selection on the maternal, paternal or embryo contributions to development are more and less likely.

Effects of ionizing irradiation and hydrostatic pressure on Escherichia coli O157:H7 inactivation, chemical composition, and sensory acceptability of ground beef patties.

A randomized complete block design with three replications was utilized to determine the effects of ionizing irradiation and hydrostatic pressure on the inactivation of Escherichia coli O157:H7, volatile composition, and consumer acceptability (n=155) of frozen ground beef patties. E-beam and X-ray irradiation (2kGy) inactivated E. coli O157:H7 below the limit of detection, while hydrostatic pressure treatment (300mPa for 5min at 4°C) did not inactivate this pathogen. Solid-phase microextraction (SPME) was used to extract volatile compounds from treated ground beef patties. Irradiation and hydrostatic pressure altered the volatile composition (P<0.05) of the ground beef patties in respect to radiolytic products. However, results were inconclusive on whether these differences were great enough to use this method to differentiate between irradiated and non-irradiated samples in a commercial setting. Irradiation did not affect (P>0.05) consumer acceptability of ground beef patties when compared to untreated samples, but hydrostatic pressure caused decreased acceptability (P<0.05) when compared to other treatments.

Characterization of fish sauce aroma-impact compounds using GC-MS, SPME-Osme-GCO, and Stevens' power law exponents.

The objectives of this study were to characterize volatile compounds and to determine the characteristic aromas associated with impact compounds in 4 fish sauces using solid-phase micro-extraction, gas chromatography-mass spectrometry, Osme, and gas chromatography olfactometry (SPME-Osme-GCO) coupled with Stevens' Power Law. Compounds were separated using GCMS and GCO and were identified with the mass spectral database, aroma perceived at the sniffing port, retention indices, and verification of compounds by authentic standards in the GCMS and GCO. Aromas that were isolated and present in all 4 fish sauce samples at all concentrations included fishy (trimethylamine), pungent and dirty socks (combination of butanoic, pentanoic, hexanoic, and heptanoic acids), cooked rice and buttery popcorn (2,6-dimethyl pyrazine), and sweet and cotton candy (benzaldehyde). All fish sauces contained the same aromas as determined by GCO and GCMS (verified using authentic standard compounds), but the odor intensity associated with each compound or group of compounds was variable for different fish sauce samples. Stevens' Power Law exponents were also determined using this analytical technique, but exponents were not consistent for the same compounds that were found in all fish sauces. Stevens' Power Law exponents ranged from 0.14 to 0.37, 0.24 to 0.34, 0.09 to 0.21, and 0.10 to 0.35 for dirty socks, fishy, buttery popcorn, and sweet aromas, respectively. This demonstrates that there is variability in Stevens' Power Law exponents for odorants within fish sauce samples.

Effect of x-ray irradiation on reducing the risk of listeriosis in ready-to-eat vacuum-packaged smoked mullet.

Listeria monocytogenes can pose a serious threat in several areas of the nation's food supply including ready-to-eat seafood products. Use of irradiation processing can potentially reduce the risk of listeriosis caused by consumption of ready-to-eat seafood products. This study measured the effect of X-ray irradiation on reducing the population of L. monocytogenes on ready-to-eat, vacuum-packaged smoked mullet. Smoked mullet were inoculated with a five-strain mixture of L. monocytogenes (10(4) CFU/g), vacuum packaged, and irradiated (0, 0.5, 1.0, 1.5, and 2.0 kGy). The packaged fish were then stored at 3 and 10 degrees C for 90 and 17 days, respectively. Radiation doses of 0.5, 1.0, and 1.5 kGy reduced the initial population of L. monocytogenes by 1.1, 1.6, and 2.1 log CFU/g, respectively. The 2.0-kGy dose reduced L. monocytogenes to undetectable levels with no recovery growth at either temperature. Compared to the control, irradiation at 1.5 kGy demonstrated 1.0 and 1.7 log CFU/g less growth at 3 degrees C after 60 days and 10 degrees C after 17 days, respectively. Sensory flavor analysis was conducted to determine if a difference existed between irradiated samples. Panelists indicated that there were no differences among treated and untreated samples. An X-ray dose of 2 kGy effectively eliminated 10(4) CFU/g L. monocytogenes on smoked mullet without changing sensory quality.

Evolutionary ecology of the prezygotic stage.

The life cycles of sexually reproducing animals and flowering plants begin with male and female gametes and their fusion to form a zygote. Selection at this earliest stage is crucial for offspring quality and raises similar evolutionary issues, yet zoology and botany use dissimilar approaches. There are striking parallels in the role of prezygotic competition for sexual selection on males, cryptic female choice, sexual conflict, and against selfish genetic elements and genetic incompatibility. In both groups, understanding the evolution of sex-specific and reproductive traits will require an appreciation of the effects of prezygotic competition on fitness.

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.

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.

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.

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.

Does differential seed siring success change over time or with pollination history in wild radish, Raphanus sativus (Brassicaceae)?

Previous work with wild radish has shown that pollen donors sire different numbers of seeds and that the condition of the maternal tissue affects seed paternity, suggesting that both pollen donor characteristics and maternal tissue affect mating. However, because these results are from the greenhouse, it is difficult to know whether they would hold true in the field. Here, we performed hundreds of crosses on several maternal plants to simulate changes during the flowering season of field plants. During the experiment, maternal resource availability changed due to the costs of producing fruits, and we determined the pollination history of a plant by performing crosses in specific orders. Examination of seed paternity showed that there were small differences in pollen donor success at the beginning of the experiment when maternal resources were abundant. Differential pollen donor success was greatest slightly later in the flowering period, but declined toward the end of the experiment. Thus, maternal plants may distinguish most among pollen donors when they have both abundant resources and experience with the differences in quality of available pollen donors. In contrast, there were few significant effects of the recent pollination history of plants on pollen donor success. Finally, despite the changes in mating performance over time, there were strong overall differences in pollen donor success, suggesting that seasonal changes in the field will not eliminate the potential for nonrandom mating.

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.

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.

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.

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.

Pollen donor performance can be consistent acrossmaternal plants in wild radish (Raphanus sativus,Brassicaceae): a necessary condition for the action of sexualselection.

The possibility that sexual selection has played a role in theevolution of plant reproductive characters remains interesting, butcontroversial. One reason is that clear demonstration of the necessaryconditions for sexual selection is lacking. For sexual selection tooccur, differences in pollen donor performance must be consistent acrossmaternal plants and not be due to mechanisms such asself-incompatibility that produce interactions between maternal plantsand pollen donors. Here, I performed two experiments with wild radish totest whether differences in pollen donor performance are consistentacross maternal plants and not due to subtle effects of theincompatibility system. In the first, all maternal and paternal lineageshad different S-alleles. There were 16 maternal plants, four in each offour lineages and four pollen donors, one in each of four lineages. Rankorder of pollen donor performance, in terms of number of seeds siredafter mixed pollination, was highly consistent across maternal plantsand maternal lineages. In addition, maternal stress treatment had aneffect on mating success of pollen donors, but the effect was subtle anddid not affect the rank order of seeds sired by the four pollen donors.In the second experiment, pollinations were performed on both mature andimmature stigmas. Immature stigmas allowed some self seed set, so theincompatibility system was compromised. There was some nonrandom seedpaternity on both mature and immature stigmas. However, the amount ofnonrandom mating was less on immature stigmas. Taken together, theseexperiments show that the kind of consistent nonrandom mating necessaryfor sexual selection occurs in wild radish, but that the incompatibilitysystem and the mechanisms for sorting among compatible mates may overlapin time of development or in somepathways.

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.

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.

Correlation of cervical lordosis measurement with incidence of motor vehicle accidents.

A retrospective analysis of 500 patient radiographs was conducted to measure the clinical correlation of cervical lordosis measurements and incidence of motor vehicle accident (MVA). Five hundred lateral cervical radiographs were selected at random from the practice of one of the authors (DLM). The C1-7 angle of the cervical curve was then measured by two blinded examiners. Inter-examiner reliability had a confidence interval of 95%. Eighty-two percent of patients who have had a MVA had an abnormal lordosis. The mean lordosis of patients who had been involved in a MVA was 26.1 degrees (SD 11.4), compared with 36.4 (SD 8.4) for those who had not been involved in a MVA. The results suggest a correlation of reduced cervical lordosis measurements following motor vehicle accident (MVA).

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.

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.