Microbial evaluation of automated sorting systems in stone fruit packinghouses during peach packing.

Automated fruit sorting systems with individual fruit carriers are utilized in modern fruit packing facilities. This study evaluated the levels of naturally occurring microflora on the surfaces of peaches and fruit carriers during automated sorting operations at stone fruit packinghouses in California. The study also assessed the growth potential of Salmonella enterica and Listeria monocytogenes on fruit carriers under various environmental conditions. No difference of microbial loads was found on peaches (n = 420) before, during, and after fruit sorting at seven packinghouses. The average surface total microbial, coliform, and yeast and mold levels of peaches during sorting were 3.6, 2.7, and 1.9 log CFU/cm2, respectively. Environmental swab testing indicated routine cleaning of fruit carriers (n = 192) reduced total microbes from 3.9 to 3.2 log CFU/cm2 (P = 0.003) and coliforms from 1.5 to 0.9 log CFU/cm2 (P = 0.001) on carriers' fruit contact surfaces. Laboratory exposures to temperature (22, 28, 34 or 40 °C) and humidity (65, 75, 85 or 95%) conditions significantly reduced inoculated Salmonella and Listeria on clean and commercially used (deposited with wax, fuzz, dirt, etc.) fruit carriers within 24 h (P < 0.001). The observed Salmonella reduction was greater on clean carriers (P < 0.001). On used carriers, Salmonella was persistent at 95% humidity and Listeria was persistent at 22 °C. The results showed the levels of surface microflora on peaches during fruit sorting, the reduction of microbial loads on fruit carriers due to packinghouses' cleaning, and the reduction, rather than growth, of Salmonella and Listeria under tested conditions on fruit carriers.

Evaluating wildlife as a potential source of Salmonella serotype Newport (JJPX01.0061) contamination for tomatoes on the eastern shore of Virginia.

Eastern Shore of Virginia red, round tomatoes contaminated with Salmonella serotype Newport pattern JJPX01.0061 have been a source of several multistate outbreaks within the last 10 years. No source of the contamination has yet been identified. The goal of this study was to evaluate wildlife as a potential source of contamination. Faecal samples from deer, turtles and birds were collected between November 2010 and July 2011 from seventeen locations on the Eastern Shore of Virginia. A total of 262 samples were tested for the presence of Salmonella using an enzyme-linked immunosorbent assay (ELISA). A total of 23 (8.8%) samples tested positive for Salmonella spp. and were further characterized by serotyping and pulsed-field gel electrophoresis (PFGE) subtyping. Overall, twelve serotypes were identified, including Salmonella serotype Javiana, another common serotype associated with tomato-related outbreaks. Only one avian sample collected in July 2011 was determined to be positive for S. Newport pattern 61. This sample was collected from the ground at a site where birds, mostly gulls, were congregating. Although many of the avian samples from this site were dry, the site yielded eleven positive Salmonella samples. This suggests that certain Salmonella serotypes may persist in the environment despite extreme conditions. The recovery of one Newport pattern 61 isolate alone does not yield much information regarding the environmental reservoirs of this pathogen, but when combined with other data including the recovery of several isolates of Javiana from birds, it suggests that birds might be a potential source of Salmonella contamination for tomatoes on the Eastern Shore.

Comparison of the microbial quality of ground beef and ground beef patties from internet and local retail markets.

This study evaluated the microbial quality of ground beef and ground beef patties sold at local (Virginia) and Internet (U.S.) retail markets. A total of 152 ground beef products, consisting of locally purchased raw ground beef (LRG) and frozen beef patties (LFP) and Internet-procured frozen ground beef (IFG) and frozen beef patties (IFP), were tested. Results showed that LFP had significantly lower levels of aerobic mesophiles, psychrotrophs, and coliforms than LRG, IFG, and IFP. Furthermore, IFG had greater numbers of Escherichia coli than LRG and LFP. No sample was contaminated with E. coli 0157: H7, but one duplicate set of summer LFP samples contained Salmonella. Listeria spp. were present in 25 and 29% of samples from local and Internet markets, respectively. About 5.0, 11.1, 10.5, and 7.9% of LRG, LFP, IFG, and IFP samples were contaminated with L. monocytogenes. This study identified differences in microbial quality between local and Internet products. Careful handling and thorough cooking of ground beef products, regardless of market source, are recommended to prevent foodborne illness.

Microbial quality of raw aquacultured fish fillets procured from Internet and local retail markets.

The microbial quality of raw fillets of aquacultured catfish, salmon, tilapia, and trout was evaluated. A total of 272 fillets from nine local and nine Internet retail markets were tested. Mean values were 5.7 log CFU/g for total aerobic mesophiles, 6.3 log CFU/g for psychrotrophs, and 1.9 log most probable number (MPN) per gram for coliforms. Differences in these microbial levels between the two kinds of markets and among the four types of fish were not significant (P > 0.05), except that Internet trout fillets had about 0.8-log higher aerobic mesophiles than did trout fillets purchased locally. Although Escherichia coli was detected in 1.4, 1.5, and 5.9% of trout, salmon, and tilapia, respectively, no sample had > or = 1.0 log MPN/g. However, E. coli was found in 13.2% of catfish, with an average of 1.7 log MPN/g. About 27% of all fillets had Listeria spp., and a positive correlation between the prevalence of Listeria spp. and Listeria monocytogenes was observed. Internet fillets had a higher prevalence of both Listeria spp. and L. monocytogenes than did those fillets purchased locally. L. monocytogenes was present in 23.5% of catfish but in only 5.7, 10.3, and 10.6% of trout, tilapia, and salmon, respectively. Salmonella and E. coli O157 were not found in any sample. A follow-up investigation using catfish operation as a model revealed that gut waste exposed during evisceration is a potential source of coliforms and Listeria spp.

Fine-scale genetic mapping of two Pierce's disease resistance loci and a major segregation distortion region on chromosome 14 of grape.

A refined genetic map of chromosome 14, which contains the Pierce's disease (PD) resistance locus, was created from three grape mapping populations. The source of PD resistance in these populations was b43-17, a male form of Vitis arizonica Engelm. that is homozygous resistant. The resistance locus segregated as a single dominant gene and mapped as PdR1a in the F1 selection F8909-17 (9621 population) and as PdR1b in a sibling F1 selection F8909-08 (04190 population). These two full sibs inherited either allele of the Pierce's disease resistance locus from the b43-17 parent, which is homozygous at that locus. The 9621 population consisted of 425 progeny and PdR1a mapped between markers VvCh14-56/VvCh14-02 and UDV095 within a 0.6 cM genetic distance. The 04190 population consisted of 361 progeny and PdR1b mapped between markers VvCh14-02 and UDV095/VvCh14-10 within a 0.4 cM distance. Many of the markers present on chromosome 14 were distorted with an excess of female alleles in the 04190 and 04373 population (developed from a cross of V. vinifera L. F2-35 x b43-17) indicating that potential gametophytic factors are present in this region. Common markers from this region within the 9621 population were not distorted except Scu15. When these markers were compared to V. vinifera-based maps of chromosome 14 they were also distorted suggesting the involvement of gametophytic factors, and prompting the identification of this region as Vitis-segregation distortion region 1 (V-SDR1). The refined genetic maps developed from this study can be used to identify and clone genes that confer resistance to Pierce's disease.

Using aqueous chlorine dioxide to prevent contamination of tomatoes with Salmonella enterica and erwinia carotovora during fruit washing.

Chlorine dioxide (ClO2) is an antimicrobial agent recognized for its disinfectant properties. In this study, the sanitizing effects of ClO2 solutions against Salmonella enterica and Erwinia carotovora in water, on tomato surfaces, and between loads of tomatoes were evaluated. In water, ClO2 at 5, 10, and 20 ppm caused a > or = 5-log reduction of S. enterica within 6, 4, and 2 s, respectively. Higher lethality was observed with E. carotovora; a 5-log reduction was achieved after only 2 s with 10 ppm ClO2. On fruit surfaces, however, the sanitizing effects were compromised. A full minute of contact with ClO2 at 20 and 10 ppm was required to achieve a 5-log reduction in S. enterica and E. carotovora counts, respectively, on freshly spot-inoculated tomatoes. On inoculated fruit surfaces, populations decreased > 3 log CFU/cm2 during desiccation at 24 +/- 1 degrees C for 24 h. Populations of air-dried Salmonella and Erwinia were not significantly reduced (P > 0.05) by ClO2 at < or = 20 ppm after 1 min. Either wet or dry inoculum of these two pathogens could contaminate immersion water, which in turn can cross-contaminate a subsequent load of clean fruit and water. ClO2 at 5 ppm used for immersion effectively prevented cross-contamination. Pathogen contamination during fruit handling is best prevented with an effective disinfectant. Once a load of fruit is contaminated with pathogens, even a proven disinfectant such as ClO2 cannot completely eliminate such contaminants, particularly when they are in a dehydrated state on fruit.

Microbial profiles of on-line--procured sprouting seeds and potential hazards associated with enterotoxigenic Bacillus spp. in homegrown sprouts.

We examined the microbiological quality of sprouting seeds sold through the Internet. Overall, five types of seeds each from six organic and six conventional sources were evaluated. The growth and toxin production of naturally occurring Bacillus spp. in sprouts produced using home-scale sprouting devices also were investigated. For alfalfa, broccoli, lentil, mungbean, and radish seeds, the average microbial counts were 3.3, 4.0, 2.8, 3.5, and 3.6 log CFU/g, presumptive B. cereus counts were 0.7, 1.0, 0.8, 1.0, and 0.9 log CFU/g, and total coliform counts were -0.3, -0.4, -0.5, 0.0, and -0.4 log of the most probable number per gram, respectively. No Salmonella, Escherichia coli O157, other fecal coliforms, or Staphylococcus aureus was found on seeds. Compared with conventional seeds, the organic seeds had lower or equivalent counts for total microorganisms, presumptive B. cereus, and total coliforms. When seeds were sprouting using a glass jar, the growth of presumptive B. cereus was significant for radish and broccoli but not for alfalfa, lentil, and mungbean sprouts; the counts exceeded 5.0 log CFU/g in radish sprouts. When sprouts were grown using an automatic sprouting device, presumptive B. cereus showed slight growth (< 3.0 log cycles) in radish, broccoli, and mungbean sprouts but no growth in alfalfa and lentil sprouts. Although the presumptive B. cereus isolates were enterotoxigenic, they did not produce or accumulate detectable levels of diarrheal toxins in freshly produced sprouts.

Microscopic observation and processing validation of fruit sanitizing treatments for the enhanced microbiological safety of fresh orange juice.

Studies were conducted to evaluate the infiltration of dye and bacteria into the interior of orange fruit and the impact of possible infiltration on achieving a 5-log microbial reduction during fresh juice processing. Fresh orange fruit were treated at the stem end area with dye and either Salmonella Rubislaw or Escherichia coli strains expressing green fluorescent protein. Microscopic images showed that bacterial contaminants localized at the surface or near surface areas that may be sanitized by surface treatments. Dye infiltration was not a reliable indicator of bacterial penetration in citrus fruit. To quantify the reduction of bacterial contamination, orange fruit were inoculated with E. coli and processed with and without hot water treatments. Greater than 5-log reductions were achieved in juice extracted from fruit immersed in hot water for 1 or 2 min at 80 degrees C, in comparison to the E. coli level detected in the control juice obtained by homogenization of inoculated fruit.

Efficacy of alkaline washing for the decontamination of orange fruit surfaces inoculated with Escherichia coli.

The effectiveness of washing treatments to decontaminate orange fruit surfaces inoculated with Escherichia coli was evaluated. Washing on roller brushes with fruit cleaners or sanitizers followed by potable water rinse reduced E. coli by 1.9 to 3.5 log cycles. Prewetting fruit for 30 s before washing provided no significant benefit in most cases. Additional sanitizing treatments either with chlorine or acid sanitizers did not enhance the results of alkaline washing. In general, high pH washing solutions (pH 11.8) applied with an adequate spray volume effectively reduced the surface contamination of fruit that lowered the microbial load of fresh juice as well.

Enhancing microbiological safety of fresh orange juice by fruit immersion in hot water and chemical sanitizers.

Trials were conducted with hot water and chemicals to sanitize Valencia oranges contaminated by natural microflora or inoculated with Escherichia coli. Microbial loads and sensory quality of fresh juice extracted from surface-heated fruit were also evaluated. E. coli on fruit surfaces was reduced by either hot water or chemical treatments. An estimated 5-log reduction of E. coli was attained by immersing inoculated fruit in hot water at 80 degrees C for 1 min or 70 degrees C for 2 min. Immersing inoculated fruit in various chemical solutions at about 30 degrees C for 8 min only reduced E. coli by about 1.8- to 3.1-log cycles on nonstem-scar surfaces of the fruit. In general, both hot water and chemical treatments were less effective at removing microflora from the stem-scar area. Rapid hot-water immersions at 80 degrees C for 1 min and 70 degrees C for 2 min reduced both fruit-surface and initial juice microbial loads without altering original sensory quality of fresh juice.

Phylogenetic characterization of novel transport protein families revealed by genome analyses.

As a result of recent genome sequencing projects as well as detailed biochemical, molecular genetic and physiological experimentation on representative transport proteins, we have come to realize that all organisms possess an extensive but limited array of transport protein types that allow the uptake of nutrients and excretion of toxic substances. These proteins fall into phylogenetic families that presumably reflect their evolutionary histories. Some of these families are restricted to a single phylogenetic group of organisms and may have arisen recently in evolutionary time while others are found ubiquitously and may be ancient. In this study we conduct systematic phylogenetic analyses of 26 families of transport systems that either had not been characterized previously or were in need of updating. Among the families analyzed are some that are bacterial-specific, others that are eukaryotic-specific, and others that are ubiquitous. They can function by either a channel-type or a carrier-type mechanism, and in the latter case, they are frequently energized by coupling solute transport to the flux of an ion down its electrochemical gradient. We tabulate the currently sequenced members of the 26 families analyzed, describe the properties of these families, and present partial multiple alignments, signature sequences and phylogenetic trees for them all.

The major facilitator superfamily.

In 1998 we updated earlier descriptions of the largest family of secondary transport carriers found in living organisms, the major facilitator superfamily (MFS). Seventeen families of transport proteins were shown to comprise this superfamily. We here report expansion of the MFS to include 29 established families as well as five probable families. Structural, functional, and mechanistic features of the constituent permeases are described, and each newly identified family is shown to exhibit specificity for a single class of substrates. Phylogenetic analyses define the evolutionary relationships of the members of each family to each other, and multiple alignments allow definition of family-specific signature sequences as well as all well-conserved sequence motifs. The work described serves to update previous publications and allows extrapolation of structural, functional and mechanistic information obtained with any one member of the superfamily to other members with limitations determined by the degrees of sequence divergence.

Reduction of microorganisms on citrus fruit surfaces during packinghouse processing.

Citrus fruit surface microbial populations were evaluated following various packingline processes of seven Florida commercial packinghouses. At each packinghouse, six fruits (oranges or tangerines) were collected at each of four sampling points. The sampling was conducted in duplicate; thus, 336 fruit were evaluated during this survey. Average aerobic plate counts and yeast and mold counts on fruit surfaces before washing were about 4.0 log CFU/cm2 and 3.3 log CFU/cm2, respectively, and were reduced to 2.1 log CFU/cm2 and 1.3 log CFU/cm2, respectively, by packinghouse processing. Waxing alone reduced the average fruit surface aerobic plate counts and coliform counts from 3.7 log CFU/cm2 and 35.2 most probable number (MPN)/cm2, respectively, to 2.6 log CFU/cm2 and 1.4 MPN/cm2. No Escherichia coli was recovered from fruit at the end of packinghouse processing, and no salmonellae were found on fruit during the entire processing. In an inoculation study to test the effect of packinghouse processes, test organism E. coli was applied to fruit to achieve a high level (4.8 log CFU/cm2) of contamination. The average E. coli count was reduced about 2.4 log cycles by washing and rinsing with potable water (40 psi, 25 degrees C) for about 30 s. The combination of washing and waxing significantly reduced the inoculated level of E. coli from 4.8 to 1.4 log CFU/cm2.

Major facilitator superfamily.

The major facilitator superfamily (MFS) is one of the two largest families of membrane transporters found on Earth. It is present ubiquitously in bacteria, archaea, and eukarya and includes members that can function by solute uniport, solute/cation symport, solute/cation antiport and/or solute/solute antiport with inwardly and/or outwardly directed polarity. All homologous MFS protein sequences in the public databases as of January 1997 were identified on the basis of sequence similarity and shown to be homologous. Phylogenetic analyses revealed the occurrence of 17 distinct families within the MFS, each of which generally transports a single class of compounds. Compounds transported by MFS permeases include simple sugars, oligosaccharides, inositols, drugs, amino acids, nucleosides, organophosphate esters, Krebs cycle metabolites, and a large variety of organic and inorganic anions and cations. Protein members of some MFS families are found exclusively in bacteria or in eukaryotes, but others are found in bacteria, archaea, and eukaryotes. All permeases of the MFS possess either 12 or 14 putative or established transmembrane alpha-helical spanners, and evidence is presented substantiating the proposal that an internal tandem gene duplication event gave rise to a primordial MFS protein prior to divergence of the family members. All 17 families are shown to exhibit the common feature of a well-conserved motif present between transmembrane spanners 2 and 3. The analyses reported serve to characterize one of the largest and most diverse families of transport proteins found in living organisms.

Evolutionary origins of multidrug and drug-specific efflux pumps in bacteria.

The available genomic sequences of three pathogenic and three nonpathogenic bacteria were analyzed to identify known and putative drug-specific and multidrug resistance transport systems. Escherichia coli was found to encode 29 such pumps, and with the exception of the archaebacterium Methanococcus jannaschii, the numbers of multidrug efflux pumps encoded within genomes of the other organisms were found to be approximately proportional to their total numbers of encoded transport systems as well as to total genome size. The similar numbers of chromosomally encoded multidrug efflux systems in pathogens and nonpathogens suggests that these transporters have not arisen recently in pathogens in response to antimicrobial chemotherapy. Phylogenetic analyses of the four transporter families that contain drug efflux permeases indicate that drug resistance arose rarely during the evolution of each family and that the diversity of current drug efflux pumps within each family arose from just one or a very few primordial systems. However, although the ability to confer drug efflux appears to have emerged on only a few occasions in evolutionary time and was stably maintained as an evolutionary trait, modulation of the substrate specificities of these systems has occurred repeatedly. A speculative model is presented that may explain the apparent capability of these multidrug transport systems to mediate drug transport from the cytoplasm or directly from the phospholipid bilayer.

[Leukopenia and hypoxemia induced by hemodialysis].

Patients with uremia usually develop hypoxemia during hemodialysis therapy. It has been thought by most of the investigators that the development of hypoxemia is related to the impropriety of biocompatibility of the dialyzer membranes. Besides, acetate dialysate through metabolism may cause a decrease of respiratory quotient and some amount of CO2 may be lost during hemodialysis. The decrease of CO2 in the blood may induce hypoventilation and therefore develop hypoxemia. A total of 32 patients who received hemodialysis therapy were studied. They were divided into group A and group B. In group A, there were 14 males and 8 females. They were hemodialyzed with cuprophan dialyzer membrane for 22 times totally. In group B, there were 6 males and 4 females. They were hemodialyzed with PMMA dialyzer membrane for 16 times totally. In both groups, acetate dialysate was used. Arterial blood gas analyses were hemodialyzed with PMMA dialyzer membrane for 16 times totally. In both groups, acetate dialysate was used. Arterial blood gas analyses were performed before hemodialysis as the baseline and at 15, 30, 45, 60, 120, 180, 240, and 300 minutes during hemodialysis. White blood cell counts (WBC) were done at the same time when arterial blood gas analyses were performed. The results showed that in group A, a marked decrease of WBC at 15 minutes (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of a low protein diet and isoenergetic amounts of a high protein diet in the weanling rat on the free amino acids of the brain.

Giving weanling rats a low protein diet for 56 days retarded the growth of the brain, whereas giving isoenergic amounts of a high protein diet did not. The low protein diet resulted in higher concentrations of aspartic acid, glycine, alanine and histidine in the plasma and of glycine and histidine in the brain. The concentration of methionine was unchanged in the plasma, but raised in the brain. Isoenergetic amounts of the high protein diet did not change the concentration of plasma amino acids, but resulted in some changes in brain amino acids. On rehabilitation the plasma and brain amino acids of the low protein animals returned to normal.