Biomedical subjects

K J Boor

Publications and source records attributed to K J Boor.

At least 19 recordsLinked to original sources

PubMed2006

Sigma B contributes to Listeria monocytogenes gastrointestinal infection but not to systemic spread in the guinea pig infection model.

Contributions of the alternative sigma factor sigmaB to Listeria monocytogenes infection were investigated using strains bearing null mutations in sigB, prfA, or inlA or in selected inlA or prfA promoter regions. The DeltaP4inlA strain, which has a deletion in the sigmaB-dependent P4inlA promoter, and the DeltasigB strain had significantly reduced invasion efficiencies relative to that of the wild-type strain in the Caco-2 human colorectal epithelial cell line, while the invasion efficiency of a strain bearing a deletion in the partially sigmaB dependent P2prfA promoter region did not differ from that of the wild type. The virulence of the DeltasigB and DeltaP4inlA strains was attenuated in intragastrically inoculated guinea pigs, with the DeltasigB strain showing greater attenuation, while the virulence capacity of the DeltaP2prfA strain was similar to that of the wild-type strain, suggesting that attenuation of virulence due to the DeltasigB mutation does not result from loss of sigmaB-dependent prfA transcription. Our results show that sigmaB-dependent activation of inlA is important for cell invasion and gastrointestinal infection and suggest that sigmaB-regulated genes in addition to inlA appear to contribute to gastrointestinal infection. Interestingly, the virulence of the DeltasigB strain was not attenuated in intravenously infected guinea pigs. We conclude that (i) L. monocytogenes sigmaB plays a critical role in invasion of human host cells, (ii) sigmaB-mediated contributions to invasion are, in part, due to direct effects on inlA transcription but not on prfA transcription, and (iii) sigmaB plays a critical role during the gastrointestinal stage of listeriosis in the guinea pig but is not important for systemic spread of the organism.

Animals↗

PubMed2004

Short communication: growth characteristics of Streptococcus uberis in UHT-treated milk.

Streptococcus uberis is an important environmental pathogen associated with bovine mastitis as well as with high total bacterial numbers in bulk tank milk. This study was conducted to determine whether S. uberis reproduction is likely to contribute to high bacterial numbers in bulk tank milk. Four S. uberis raw milk isolates were individually inoculated into UHT-treated milk and incubated at 4.4 or 7 degrees C for up to 5 d to simulate appropriate cooling; at 10 degrees C for 5 d to simulate marginally inadequate cooling; at 21 or 25 degrees C for 7 h to simulate ambient temperatures; or at 32 degrees C for 7 h to simulate elevated temperature conditions. None of the S. uberis isolates grew at either 4.4 or 7 degrees C. Streptococcus uberis growth at 10 degrees C appeared to be ribotype-specific. Although ribotype 116-520-S-1 isolates did not grow at 10 degrees C, ribotype 116-520-S-2 isolate numbers increased up to 3.5 log10 cfu/mL within 5 d. Generation times were calculated as 2.7 +/- 0.1 h, 2.1 +/- 0.1 h, and 1.0 +/- 0.1 h for 116-520-S-1 isolates and 1.8 +/- 0.4 h, 1.3 +/- 0.3 h, and 0.8 +/- 0.1 h for 116-520-S-2 isolates at 21, 25, and 32 degrees C, respectively. Our results suggest that high numbers of S. uberis in bulk tank milk are more likely to reflect high numbers of S. uberis shed by mastitic cows, rather than multiplication of these organisms under cooling conditions required for production of Grade A milk.

Animals↗

PubMed2004

Molecular Subtyping and Tracking of Listeria monocytogenes in Latin-Style Fresh-Cheese Processing Plants.

Latin-style fresh cheeses, which have been linked to at least 2 human listeriosis outbreaks in the United States, are considered to be high-risk foods for Listeria monocytogenes contamination. We evaluated L. monocytogenes contamination patterns in 3 Latin-style fresh-cheese processing plants to gain a better understanding of L. monocytogenes contamination sources in the manufacture of these cheeses. Over a 6-mo period, 246 environmental samples were collected and analyzed for L. monocytogenes using both the Food and Drug Administration (FDA) method and the Biosynth L. monocytogenes detection system (LMDS). Finished cheese samples from the same plants (n = 111) were also analyzed by the FDA method, which was modified to include L. monocytogenes plating medium (LMPM) and the L. monocytogenes confirmatory plating medium (LMCM) used in the LMDS method. Listeria monocytogenes was detected in 6.3% of cheese and 11.0% of environmental samples. Crates, drains, and floor samples showed the highest contamination rates, with 55.6, 30.0, and 20.6% L. monocytogenes positive samples, respectively. Finished products and food contact surfaces were positive in only one plant. The FDA method showed a higher sensitivity than the LMDS method for detection of L. monocytogenes from environmental samples. The addition of LMPM and LMCM media did not further enhance the performance of the FDA method for L. monocytogenes detection from finished products. Molecular subtyping (PCR-based allelic analysis of the virulence genes actA and hly and automated ribotyping) was used to track contamination patterns. Ribotype DUP-1044A, which had previously been linked to a 1998 multistate human listeriosis outbreak in the United States, was the most commonly identified subtype (20/36 isolates) and was isolated from 2 plants. This ribotype was persistent and widespread in one factory, where it was also responsible for the contamination of finished products. We hypothesize that this ribotype may represent a clonal group with a specific ability to persist in food processing environments. While previous listeriosis outbreaks were linked to Latin-style fresh cheeses made from unpasteurized milk, the presence of this organism in pasteurized cheese products illustrates that persistent environmental contamination also represents an important source of finished product contamination.

Cheese↗

PubMed2004

Mastitis-causing streptococci are important contributors to bacterial counts in raw bulk tank milk.

The objective of this study was to probe the contribution of streptococci to the microbial quality of raw milk. Over a 5-month period, bulk tank milk samples from 48 New York State dairy farms were analyzed qualitatively for bacterial ecology and quantitatively for total bacterial, streptococcal, staphylococcal, and gram-negative bacterial counts. Linear regression analysis was used to determine the contribution of differential counts to total bacterial counts. Streptococci, staphylococci, and gram-negative bacteria accounted for 69, 3, and 3% of total bacterial count variability, respectively. Randomly selected Streptococcus isolates from each bulk tank milk sample were identified to species by means of the API 20 STREP identification system. The most commonly identified streptococcal species were Streptococcus uberis, Aerococcus viridans, and Streptococcus agalactiae, which were detected in 81, 50, and 31% of 48 bulk tank samples, respectively. For five herds, S. uberis isolates from bulk tank milk and individual cows were characterized by PvuII ribotyping. A farm-specific dominant ribotype was found in each bulk tank sample, and that ribotype was isolated from at least one cow within each herd of origin. Bacteriological and strain typing data indicate that control of streptococci, specifically mastitis-causing species, is important for improvement of the microbial quality of raw milk in New York State.

Animals↗

PubMed2002

Vitamin A degradation and light-oxidized flavor defects in milk.

To determine the effects of light exposure on vitamin A degradation and on light-oxidized flavor development, samples of whole, reduced fat, and nonfat milk were exposed to fluorescent light (either 1000 or 2000 lx) at time intervals of 2, 4, 8, or 16 h. Measurable vitamin A losses occurred at 2, 4, and 16 h at 2000 lx for nonfat, reduced fat and whole milk, respectively. Moderate light-oxidized flavors were detected after 4 h of light exposure (2000 lx) in the whole and reduced fat milk and after 8 h in nonfat milk. The different types of milk show a significant difference in relative flavor scores. By 16 h at 2000 lx, relative light-oxidized flavor development was lower in nonfat milk than in whole or reduced fat milk. The presence of milk fat appears to protect against vitamin A degradation in fluid products, but adversely affects the flavor quality of milk after exposure to light. In summary, these findings demonstrate that even a brief, moderate light exposure (2 h; 2000 lx) can reduce the nutritional value and flavor quality of fluid milk products.

Animals↗

PubMed2001

Molecular studies on the ecology of Listeria monocytogenes in the smoked fish processing industry.

We have applied molecular approaches, including PCR-based detection strategies and DNA fingerprinting methods, to study the ecology of Listeria monocytogenes in food processing environments. A total of 531 samples, including raw fish, fish during the cold-smoking process, finished product, and environmental samples, were collected from three smoked fish processing facilities during five visits to each facility. A total of 95 (17.9%) of the samples tested positive for L. monocytogenes using a commercial PCR system (BAX for Screening/Listeria monocytogenes), including 57 (27.7%) environmental samples (n = 206), 8 (7.8%) raw material samples (n = 102), 23 (18.1%) samples from fish in various stages of processing(n = 127), and 7 (7.3%) finished product samples (n = 96). L. monocytogenes was isolated from 85 samples (16.0%) using culture methods. Used in conjunction with a 48-h enrichment in Listeria Enrichment Broth, the PCR system had a sensitivity of 91.8% and a specificity of 96.2%. To track the origin and spread of L. monocytogenes, isolates were fingerprinted by automated ribotyping. Fifteen different ribotypes were identified among 85 isolates tested. Ribotyping data established possible contamination patterns, implicating raw materials and the processing environment as potential sources of finished product contamination. Analysis of the distribution of ribotypes revealed that each processing facility had a unique contamination pattern and that specific ribotypes persisted in the environments of two facilities over time (P < or = 0.0006). We conclude that application of molecular approaches can provide critical information on the ecology of different L. monocytogenes strains in food processing environments. This information can be used to develop practical recommendations for improved control of this important food-borne pathogen in the food industry.

Animals↗

PubMed2001

Role of sigma(B) in heat, ethanol, acid, and oxidative stress resistance and during carbon starvation in Listeria monocytogenes.

To determine the contribution of sigma B (sigma(B)) to survival of stationary-phase Listeria monocytogenes cells following exposure to environmental stresses, we compared the viability of strain 10403S with that of an isogenic nonpolar sigB null mutant strain after exposure to heat (50 degrees C), ethanol (16.5%), or acid (pH 2.5). Strain viabilities were also determined under the same conditions in cultures that had been previously exposed to sublethal levels of the same stresses (45 degrees C, 5% ethanol, or pH 4.5). The DeltasigB and wild-type strains had similar viabilities following exposure to ethanol and heat, but the DeltasigB strain was almost 10,000-fold more susceptible to lethal acid stress than its parent strain. However, a 1-h preexposure to pH 4.5 yielded a 1,000-fold improvement in viability for the DeltasigB strain. These results suggest the existence in L. monocytogenes of both a sigma(B)-dependent mechanism and a pH-dependent mechanism for acid resistance in the stationary phase. sigma(B) contributed to resistance to both oxidative stress and carbon starvation in L. monocytogenes. The DeltasigB strain was 100-fold more sensitive to 13.8 mM cumene hydroperoxide than the wild-type strain. Following glucose depletion, the DeltasigB strain lost viability more rapidly than the parent strain. sigma(B) contributions to viability during carbon starvation and to acid resistance and oxidative stress resistance support the hypothesis that sigma(B) plays a role in protecting L. monocytogenes against environmental adversities.

Adaptation, Physiological↗

PubMed2001

Characterization and pathogenic potential of Listeria monocytogenes isolates from the smoked fish industry.

This study was designed to evaluate the hypothesis that some of the Listeria monocytogenes subtypes associated with foods, specifically smoked fish, may have an attenuated ability to cause human disease. We tested this hypothesis by using two different approaches: (i) comparison of molecular subtypes found among 117 isolates from smoked fish, raw materials, fish in process, and processing environments with subtypes found among a collection of 275 human clinical isolates and (ii) the evaluation of the cytopathogenicity of industrial isolates. Ribotyping and PCR-restriction fragment length polymorphism typing of the hlyA and actA genes differentiated 23 subtypes among the industrial isolates and allowed classification of the isolates into three genetic lineages. A significantly higher proportion of human isolates (69.1%) than industrial isolates (36.8%) were classified as lineage I, which contains human sporadic isolates and all epidemic isolates. All other industrial isolates (63.2%) were classified as lineage II, which contains only human sporadic isolates. Lineage I ribotypes DUP-1038B and DUP-1042B represented a significantly higher proportion of the human isolates than industrial isolates (5.1%). Lineage II ribotypes DUP-1039C, DUP-1042C, and DUP-1045, shown previously to persist in the smoked fish processing environment, represented nearly 50% of the industrial isolates, compared to 7.6% of the human isolates. Representatives of each subtype were evaluated with a tissue culture plaque assay. Lineage I isolates formed plaques that were significantly larger than those formed by lineage II isolates. Isolates from the smoked fish industry representing three ribotypes formed no plaques or small plaques, indicating that they had an impaired ability to infect mammalian cells. While L. monocytogenes clonal groups linked to human listeriosis cases and outbreaks were isolated, our data also suggest that at least some L. monocytogenes subtypes present in ready-to-eat foods may have limited human-pathogenic potential.

Animals↗

PubMed2001

Fluid dairy product quality and safety: looking to the future.

The fiercely competitive nature of the US beverage industry will drive the fluid milk sector of the dairy industry to improve product quality and shelf life to enable dairy beverages to compete with innovative new introductions as well as with currently popular shelf-stable products. The recent substantial growth in the volume of flavored milk sales specifically suggests that attention is needed to improve these products. Further, increasing public awareness and regulatory attention directed toward food safety issues highlight the need for the dairy industry to proactively address and eliminate emerging food safety issues that may negatively impact the image of dairy products. Shelf life and sensory profiles of high temperature short time pasteurized fluid milk products are presented, illustrating the need for greater attention to controlling contaminating microorganisms in processed fluid milk products. Bacterial spoilage patterns of flavored versus unflavored milks are compared, and suggestions are presented for extending flavored product shelf lives. Strategies currently applied to extend shelf life are reviewed. Food safety issues facing the dairy industry are presented within the context of an overview of foodborne illnesses in the United States. The pressing need to determine thermal resistance characteristics of Mycobacterium paratuberculosis is described.

Animals↗

PubMed2001

Quantitative descriptive analysis and principal component analysis for sensory characterization of ultrapasteurized milk.

Quantitative descriptive analysis was used to describe the key attributes of nine ultrapasteurized (UP) milk products of various fat levels, including two lactose-reduced products, from two dairy plants. Principal components analysis identified four significant principal components that accounted for 87.6% of the variance in the sensory attribute data. Principal component scores indicated that the location of each UP milk along each of four scales primarily corresponded to cooked, drying/lingering, sweet, and bitter attributes. Overall product quality was modeled as a function of the principal components using multiple least squares regression (R2 = 0.810). These findings demonstrate the utility of quantitative descriptive analysis for identifying and measuring UP fluid milk product attributes that are important to consumers.

Animals↗

PubMed2001

Identification and characterization of elevated microbial counts in bulk tank raw milk.

The bacterial composition of bulk tank milk from 13 farms was examined over a 2-wk period to characterize sudden elevations in the total bacterial count referred to as "spikes." Bulk tank milk samples collected at each pick-up were analyzed for standard plate count, Petrifilm aerobic count, somatic cell count, gram-negative organisms, and streptococci. Twenty standard plate count spikes were observed: 12 associated with streptococci, 4 associated with gram-negative organisms, 2 associated with streptococci and gram-negative organisms, and 2 that were not definitively characterized. Spikes ranged from 14,000 to 600,000 cfu/ml. Streptococcus uberis was isolated as the predominant organism from 11 spikes, and Escherichia coli was isolated from 4 spikes. Statistical analysis of total bacterial counts indicated a high correlation (r = 0.94) between standard plate counts and Petrifilm aerobic count. Regression analysis of standard plate counts and Petrifilm aerobic counts yielded the equation log10 (standard plate count) = 0.73 + 0.85log10 (Petrifilm aerobic count), indicating that the correlation, although strong, is not one to one. In a related pilot study, triplicate bulk tank milk samples were collected and analyzed for total bacterial count and presumptive streptococcus, gram-negative, and staphylococcus counts. Two-way ANOVA of these triplicate data indicated a lack of significant variation among the triplicate samples, suggesting that one sample can reliably gauge the microbial status of the entire bulk tank.

Animals↗

PubMed2001

Acceptance of 2% ultra-pasteurized milk by consumers, 6 to 11 years old.

The objective of this study was to determine the degree of liking of ultra-pasteurized (UP) milk by 6 to 11 year olds. For comparative purposes, 2% UP milks were evaluated along with 2% conventionally pasteurized high temperature short time (HTST) and 2% ultra high temperature (UHT) milks. A seven-point facial hedonic scale with Peryam & Kroll verbal descriptors for affective testing with children was used with the 6 year olds. For the older children, a seven-point hedonic scale with Peryam & Kroll verbal descriptors was used. The mean degree of liking of UP milk was rated as slightly below "good." HTST milk was liked slightly more than the UHT milk, which was liked slightly more than the UP milk. How the children felt about milk, in general, significantly affected how much they liked the test milks, with, all milks being influenced equally.

Animals↗

PubMed2001

Fluid milk vitamin fortification compliance in New York State.

Current US regulations, as specified in the Pasteurized Milk Ordinance, require vitamin A fortification of all reduced fat fluid milk products. The addition of vitamin D is optional in all fluid products. Acceptable vitamin concentrations in fortified milks are 2000 to 3000 International units per quart for vitamin A and 400 to 600 International units per quart for vitamin D. Vitamin A and D levels were analyzed in fortified milk products collected over a 4-yr period in New York State. Samples of whole fat, 2% fat, 1% fat, and nonfat milks were collected twice per year from up to 31 dairy processing plants. For vitamin A, 44.5% of 516 samples were in compliance with current regulations, and 47.7% of 648 samples were within the acceptable range for vitamin D. Most milk samples that were out of compliance were underfortified.

Animals↗

PubMed2000

Molecular and phenotypic characterization of Pseudomonas spp. isolated from milk.

Putative Pseudomonas spp. isolated predominantly from raw and processed milk were characterized by automated ribotyping and by biochemical reactions. Isolates were biochemically profiled using the Biolog system and API 20 NE and by determining the production of proteases, lipases, and lecithinases for each isolate. Isolates grouped into five coherent clusters, predominated by the species P. putida (cluster A), P. fluorescens (cluster B), P. fragi (as identified by Biolog) or P. fluorescens (as identified by API 20 NE) (cluster C), P. fragi (as identified by Biolog) or P. putida (as identified by API 20 NE) (cluster D), and P. fluorescens (cluster E). Isolates within each cluster also displayed similar enzyme activities. Isolates in clusters A, C, and D were generally negative for all three enzyme activities; isolates in cluster B were predominantly positive for all three enzyme activities; and isolates in cluster E were negative for lecithinase but predominantly positive for protease and lipase activities. Thus, only isolates from clusters B and E produced enzyme activities associated with dairy product flavor defects. Thirty-eight ribogroups were differentiated among the 70 isolates. Ribotyping was highly discriminatory for dairy Pseudomonas isolates, with a Simpson's index of discrimination of 0.955. Isolates of the same ribotype were never classified into different clusters, and ribotypes within a given cluster generally showed similar ribotype patterns; thus, specific ribotype fragments may be useful markers for tracking the sources of pseudomonads in dairy production systems. Our results suggest that ribogroups are generally homogeneous with respect to nomenspecies and biovars, confirming the identification potential of ribotyping for Pseudomonas spp.

Animals↗

PubMed2000

Effects of somatic cell count on quality and shelf-life of pasteurized fluid milk.

Milk was collected from eight Holstein cows four times before and four times after intramammary infection with Streptococcus agalactiae. Postinfection milk had significantly higher somatic cell count (SCC) (849,000 cells/ml) than preinfection milk (45,000 cells/ml). High SCC raw milk had more lipolysis and proteolysis than low SCC raw milk. Pasteurized, homogenized, 2% fat milks from pre- and postinfection periods were stored at 5 degrees C and analyzed for lipolysis, proteolysis, microbial quality, and sensory attributes at 1, 7, 14, and 21 d post processing. During refrigerated storage, the average rates of free fatty acid increase (i.e., lipolysis) and casein hydrolysis in high SCC milk were, respectively, three and two times faster than those in low SCC milk. In general, standard plate counts, coliform counts, and psychrotrophic bacterial counts of both the high and low SCC milks remained low (<100,000 cfu/ ml) during 5 degrees C storage. Low SCC milk maintained high organoleptic quality for the entire 21-d shelf-life period. However, for high SCC milk, between 14 and 21 d, sensory defects were detected, which resulted in low overall quality ratings. The sensory defects mainly included rancidity and bitterness and were consistent with higher levels of lipolysis and proteolysis. Hence, mastitis adversely affected the quality of pasteurized fluid milk. It is recommended that the fluid milk industry consider implementation of premium quality payment programs for low SCC milks.

Animals↗

PubMed2000

Application of the BAX for screening/genus Listeria polymerase chain reaction system for monitoring Listeria species in cold-smoked fish and in the smoked fish processing environment.

The cold-smoked fish industry was used as a model for the development of a system for monitoring Listeria spp. in foods and in the food processing environment. A total of 214 samples including raw fish, fish during the cold-smoking process, finished product, and environmental samples were collected from three processing facilities over two visits to each facility. Samples were screened for Listeria spp. using the BAX for Screening/genus Listeria polymerase chain reaction system (PCR) and by culture. Listeria spp., confirmed by the API Listeria test strip or by a PCR assay targeting the L. monocytogenes hlyA gene, were isolated from a total of 89 (41.6%) samples. Of these, 80 samples also tested positive for Listeria spp. using the BAX system. Specifically, 42 (55.3%) environmental samples (n = 76), 11 (25.6%) raw materials samples (n = 43), 20 (35.1%) samples from fish in various stages of processing (n = 57), and 7 (18.4%) finished product samples (n = 38) tested positive for Listeria spp. using the BAX system. Five (4.0%) of the 125 culture-negative samples yielded BAX system-positive results. Listeria isolates from each of nine culture-positive/BAX system-negative samples yielded a positive reaction when tested in pure culture by the BAX system, suggesting that our false-negative results were likely due to the presence of low Listeria numbers in the initial enrichment as opposed to nonreacting isolates. The employment of alternative enrichment protocols, such as the two-step enrichment recommended by the manufacturer, may increase the sensitivity of the assay.

Animals↗

PubMed2000

Characterization of chocolate milk spoilage patterns.

Standard plate counts (SPC) and psychrotrophic plate counts (PPC) from chocolate milk samples were compared with those of unflavored milk samples plated within 24 h of processing and at 7, 10, and 14 days of storage at 6 degrees C using matched samples collected over four time periods from four milk-processing plants. Bacterial numbers within 24 h of processing were not significantly different in unflavored and in chocolate milk samples (P > 0.001), with SPC less than 1,000 CFU/ml and PPC below 10 CFU/ml for both types of products. SPC and PPC were higher in chocolate milk samples than in unflavored milk samples collected from all four plants after 14 days of storage (P < 0.001). To examine the effects of chocolate milk components on bacterial numbers, SPC for days 0, 7, 14, and 21 were monitored in samples of experimentally prepared unflavored milk, milk with chocolate powder and sucrose (chocolate milk), milk with sucrose only, and milk containing chocolate powder only. At days 14 and 21, SPC were higher in both chocolate milk and in milk with chocolate powder only, than in either the unflavored milk or milk with sucrose only (P < 0.001). These findings suggest that the addition of chocolate powder to milk can contribute to a greater relative increase in bacterial numbers in pasteurized chocolate milk than in identically processed unflavored milk at 14 days postprocessing.

Animals↗

PubMed1999

Molecular investigation of a listeriosis outbreak in goats caused by an unusual strain of Listeria monocytogenes.

During a 16-month period, 10 goats with listeriosis were identified in 2 herds that shared 3 bucks, including 1 that died of listeriosis. Using DNA fingerprinting, we determined that a single genetically unique Listeria monocytogenes strain had infected all goats from which isolates were available. All isolates were unable to metabolize rhamnose (rhamnose-negative), whereas as a species, L monocytogenes is considered to have a rhamnose-positive phenotype. Therefore, these isolates would have been characterized as a species other than L monocytogenes if any of a variety of commercial bacterial identification kits had been used for speciation. Silage was not fed to either herd, and L monocytogenes was not isolated from vaginal or rectal swab specimens obtained from healthy goats or from samples of feed. Because the 3 bucks were the only common elements between the 2 herds, our results suggest a venereal route of transmission for listeriosis.

Abortion, Veterinary↗