Incidence and relationship of group D streptococci with other indicator organisms in meats.

Raw and processed meats were analyzed for presumptive group D streptococci using KF streptococcus agar. Counts were compared with coliform, presumptive Escherichia coli, and Enterobacteriaceae counts but no meaningful relationships were observed. Results indicated that group D streptococci and E. coli type I were principally contaminants from the packing plant, rather than at retail level. The predominating group D streptococcus in both beef and pork cuts was Streptococcus faecalis, while in processed meat (bologna), the predominating group D streptococci were Streptococcus faecium var. durans and Streptococcus faecium. Streptococcus bovis was not detected among the isolates from any meat samples. Marked differences were noted in numbers of group D streptococci in processed meat from different manufacturers. The results did not support the use of group D streptococci as alternative indicator organisms for meats. However, the association of group D streptococci with packing plant contamination may prove to be of value.     

MEMBRANE LIFTING TECHNIQUE FOR RAPID DETECTION OF ESCHERICHIA COLI 0157:H7 IN INOCULATED GROUND MEATS

A membrane lifting technique was developed for direct rapid detection of Escherichia coli 0157:H7 in inoculated ground meats. Duplicate groups of 2 meatballs were inoculated with volumes of 0.1-ml of a serial dilution (1:10) of E. coli 0157:H7 or a mixed culture containing one strain of E. coli 0157:H7 and a non-0157:H7E. coli serotype (E. coli ATCC 25922). Each meatball was sampled by sandwiching with 2 pieces of nitrocellulose membranes and pressing against each other to the center of the meatball. The membranes were in contact with the meats for 10 min to lift the bacteria. The membranes were removed and incubated on MacConkey-sorbitol agar plates with the meat contact side up. After 18 h incubation at 37C, an immunostain was performed directly on the membranes for detection of the presence or absence of E. coli 0157:H7. This method was found to be sensitive enough to detect as few as 1 to 2 cells of E. coli 0157:H7 inoculated on surfaces of 18-g meatballs. This method might be used as a rapid presumptive test for E. coli 0157:H7.     

Improved walnut-feeding skills with experience in wood mice, <Emphasis Type="Italic">Apodemus speciosus</Emphasis>

The wood mouse Apodemus speciosus can consume the hard-walled walnut species Juglans ailanthifolia. The mice gnaw holes on two sides of the shell and then pick the meat from the holes. However, not all mice are able to eat these walnuts, because the shells are extremely hard and the process is labour-intensive. To consume all of the meat it is more efficient to eat from holes on the raphe than to attempt to eat from other parts of the shell. We examined the effect of experience on the walnut-feeding skills of mice in the field. Feeding behaviours were compared among mice from habitats with and without walnut trees. Mice from habitats with walnut trees tended to consume nuts more efficiently than mice from habitats without walnut trees. We also observed the feeding behaviour of mice in an experimental area into which walnuts were artificially placed over a period of one year. This manipulation increased the proportion of mice that were able to eat nuts frequently and efficiently. Therefore, the walnut-feeding skills of mice improved with experience. Because individual mice acquired efficient feeding during the 14-day period of walnut conditioning, trial-and-error learning may be an important mechanism contributing to this behaviour.     

Depression of lymphocyte transformation and exacerbation of Behcet's syndrome by ingestion of english walnuts

Extracts of English and black walnuts, almonds and pecans stimulated increased incorporation of tritiated thymidine by cultured leucocytes from both normals and patients with Behcet's syndrome. English walnut extract also stimulated newborn cord blood leucocytes, indicating it to be a nonspecific stimulant. Within 2 days after ingestion of English walnuts by normals and patients with Behcet's syndrome, their in vitro lymphocyte reactivity to both the walnut extract and Candida antigens was significantly decreased. This inhibition of lymphocyte transformation may be more prolonged in the patients than in normal subjects and was associated with an increase in frequency and severity of their symptoms. Hypotheses relating the mechanism of action of these two apparently disparate phenomenon are discussed.     

Comparison of the prevalences and antimicrobial resistances of Escherichia coli isolates from different retail meats in the United States, 2002 to 2008

Escherichia coli isolates were recovered from the National Antimicrobial Resistance Monitoring System retail meat program and examined for antimicrobial susceptibility. Retail meat samples (n = 11,921) from four U.S. states collected during 2002 to 2008, consisting of 2,988 chicken breast, 2,942 ground turkey, 2,991 ground beef, and 3,000 pork chop samples, were analyzed. A total of 8,286 E. coli isolates were recovered. The greatest numbers of samples contaminated with the organism were chicken (83.5%) and turkey (82.0%), followed by beef (68.9%) and pork (44.0%). Resistance was most common to tetracycline (50.3%), followed by streptomycin (34.6%), sulfamethoxazole-sulfisoxazole (31.6%), ampicillin (22.5%), gentamicin (18.6%), kanamycin (8.4%), amoxicillin-clavulanic acid (6.4%), and cefoxitin (5.2%). Less than 5% of the isolates had resistance to trimethoprim, ceftriaxone, ceftiofur, nalidixic acid, chloramphenicol, and ciprofloxacin. All isolates were susceptible to amikacin. Compared to beef and pork isolates, the poultry meat isolates had a greater percentage of resistance to all tested drugs, with the exception of chloramphenicol, to which pork isolates had the most resistance. More than half of the turkey isolates (56%) were resistant to multidrugs (≥3 classes) compared to 38.9% of chicken, 17.3% of pork, and 9.3% of beef isolates. The bla(CMY) gene was present in all ceftriaxone- and ceftiofur-resistant isolates. The cmlA, flo, and catI genes were present in 45%, 43%, and 40% of chloramphenicol-resistant isolates, respectively. Most nalidixic acid-resistant isolates (98.5%) had a gyrA mutation in S83 or D87 or both, whereas only 6.7% had a parC mutation in either S80 or E84. The results showed that E. coli was commonly present in the retail meats, and antimicrobial resistance profiles differed according to the animal origin of the isolates.     

Detection of walnut residues in processed foods by polymerase chain reaction

A sensitive qualitative detection method for walnut (Juglans regia) using polymerase chain reaction (PCR) was developed. For detection of walnuts with high specificity, the primer pair WAL-F/WAL-R was designed based on walnut matK genes. Trace amounts of walnuts in commercial food products can be qualitatively detected using this method.     

Exposure to non-acid fresh meat decontamination washing fluids sensitizes Escherichia coli O157:H7 to organic acids

AIMS: To investigate whether Escherichia coli O157:H7 maintains acid tolerance in water meat decontamination washing fluids. METHODS AND RESULTS: A rifampicin-resistant derivative of E. coli O157:H7 strain ATCC 43895 was inoculated (10(5) cfu ml(-1)) in spray-washings from meat sprayed with cold (10 degrees C) or hot (85 degrees C) water, stored at 10 degrees C for up to 14 days, and its acid tolerance was assessed at 2 and 8 days by exposure to broth or new washings adjusted to pH 3.5 or 3.7 with lactic or acetic acid. The pathogen survived in the water washings, but it was outgrown by the natural, Pseudomonas-like flora, and it was sensitized to acid. CONCLUSIONS: The acid tolerance of E. coli O157:H7 decreases following exposure to non-acid, but otherwise stressful, conditions prevailing in water meat washings at 10 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings suggest that the more intense use of water-based technologies should be included in meat decontamination strategies because they may contribute to enhanced meat safety by inducing acid sensitization in E. coli O157:H7.     

Alfalfa N2-fixation enhances the phosphorus uptake of walnut in interplantings

Walnut tree seedlings exhibited greater phosphorus (P) uptake from³²P-labelled hydroxyapatite when interplanted with alfalfa than with other walnuts, black locust, or orchard grass. Three mechanisms are proposed as possible explanations of this enhnaced P uptake by walnut. In this study, diffusion of solubilized apatite-P to the roots of walnut at points of walnut-alfalfa root intersection is believed to be the operative mechanism. Phosphorus is solubilized due to rhizosphere acidification of alfalfa during nitrogen fixation. These results underscore the interdependence of nutrient cycles. Enhancement of the phosphorus cycle through manipulation of the nitrogen cycle has important implications for world food and fiber production.     

Comparison of EC broth and medium A-1 for the recovery of Escherichia coli from frozen shucked snow crab.

Two variations of the multiple-tube fermentation technique were used to enumerate fecal coliforms in commercially processed, frozen crab meat. These were the EC confirmation test and a more rapid method that requires medium A-1. The method with medium A-1 was more specific than the EC confirmation test for detecting Escherichia coli type 1. E. coli was isolated from 84% of the positive medium A-1 tubes, whereas it was isolated from only 64% of the positive tubes of EC broth. When samples of crab meat were inoculated with known amounts of E. coli, better estimates of the known numbers were obtained by the medium A-1 method. Several species of nonfecal coliforms were isolated from cultures in EC broth. These belonged to the genera Klebsiella, Citrobacter, Enterobacter, and Serratia. Apparently these strains were naturally adapted to growth at an elevated temperature because the majority were able to grow at 44.5 degrees C when retested in EC broth. Fewer species of nonfecal coliforms were isolated from medium A-1. Those that were isolated belonged to the genera Citrobacter and Enterobacter.     

Comparison of EC broth and medium A-1 for the recovery of Escherichia coli from frozen shucked snow crab.

Two variations of the multiple-tube fermentation technique were used to enumerate fecal coliforms in commercially processed, frozen crab meat. These were the EC confirmation test and a more rapid method that requires medium A-1. The method with medium A-1 was more specific than the EC confirmation test for detecting Escherichia coli type 1. E. coli was isolated from 84% of the positive medium A-1 tubes, whereas it was isolated from only 64% of the positive tubes of EC broth. When samples of crab meat were inoculated with known amounts of E. coli, better estimates of the known numbers were obtained by the medium A-1 method. Several species of nonfecal coliforms were isolated from cultures in EC broth. These belonged to the genera Klebsiella, Citrobacter, Enterobacter, and Serratia. Apparently these strains were naturally adapted to growth at an elevated temperature because the majority were able to grow at 44.5 degrees C when retested in EC broth. Fewer species of nonfecal coliforms were isolated from medium A-1. Those that were isolated belonged to the genera Citrobacter and Enterobacter.     

Changes in Escherichia coli O157:H7 numbers during holding on excised lean, fascia and fat beef surfaces at different temperatures

Aim:  To investigate changes in Escherichia coli O157:H7 numbers on excised beef carcass surfaces over 72 h at different temperatures.
Methods and Results:  Excised lean meat, fascia and fat were inoculated with E. coli O157:H7 and held in an environmental chamber for 72 h, at air speed 0·5 m s⁻¹, relative humidity (RH) 90%, and temperatures 4, 8 and 12°C. On lean, pathogen counts increased significantly at 12°C. On fascia, significant reductions in counts occurred at 4 and 8°C. Pathogen numbers were significantly reduced on fat at 4, 8 and 12°C (64 h). Counts on fat were significantly less at all temperatures, compared to lean or fascia and surface water activity, a_w, decreased significantly over time on fat at 4°C. Significant decreases in surface pH values were recorded on all meat substrates.
Conclusions:  The survival of E. coli O157:H7 varied in relation to the meat substrate and the holding temperature. Reductions in counts on fat surfaces appeared to be related to low surface a_w values. No relationship between pathogen survival and surface pH was established.
Significance and Impact of the Study:  The use of excised meat pieces in an environmental cabinet offers a more flexible approach to determining the use of different chilling regimes in the production of safe meat.     

Historical effects on local variation in walnut-feeding behavior by the Japanese wood mouse, Apodemus speciosus

The Japanese wood mouse Apodemus speciosus eats large, hard-walled walnuts of Juglans ailanthifolia immediately after finding or after hoarding them. However, not all individuals can efficiently eat the nuts. In this study, to examine local variation in the ability to eat walnuts, feeding behavior was compared among nine wood mouse populations, four from mainland Honshu, where the walnut tree is distributed, and five from the Izu Islands (30-100 km south of Honshu), which lack the tree species. To avoid the effects of pre-capture experience with walnuts, mice from areas lacking the walnut trees were used for testing, even in the Honshu sites. Most mice from Honshu were able to eat walnuts after a 14-day training period, whereas most insular mice could not, with the exception of mice on Kouzushima Island. An analysis of the population genetic structures of these mice based on sequences of the mitochondrial control (D-loop) region revealed that the four insular populations are genetically distinct from the mainland populations, whereas the Kouzushima population remains genetically similar to the mainland populations. The relatively recent colonization of Kouzushima may explain why mice from this island were able to feed on walnuts despite the lack of walnut trees on the island. Thus, walnut-feeding ability appears to have some innate basis in the Japanese wood mouse, and this trait would be selected for in a walnut-available environment as it would better enable mice to survive during food shortages.     

Prevalence of Campylobacter spp., Escherichia coli, and Salmonella serovars in retail chicken, turkey, pork, and beef from the Greater Washington, D.C., area.

A total of 825 samples of retail raw meats (chicken, turkey, pork, and beef) were examined for the presence of Escherichia coli and Salmonella serovars, and 719 of these samples were also tested for Campylobacter spp. The samples were randomly obtained from 59 stores of four supermarket chains during 107 sampling visits in the Greater Washington, D.C., area from June 1999 to July 2000. The majority (70.7%) of chicken samples (n = 184) were contaminated with Campylobacter, and a large percentage of the stores visited (91%) had Campylobacter-contaminated chickens. Approximately 14% of the 172 turkey samples yielded Campylobacter, whereas fewer pork (1.7%) and beef (0.5%) samples were positive for this pathogen. A total of 722 Campylobacter isolates were obtained from 159 meat samples; 53.6% of these isolates were Campylobacter jejuni, 41.3% were Campylobacter coli, and 5.1% were other species. Of the 212 chicken samples, 82 (38.7%) yielded E. coli, while 19.0% of the beef samples, 16.3% of the pork samples, and 11.9% of the turkey samples were positive for E. coli. However, only 25 (3.0%) of the retail meat samples tested were positive for Salmonella. Significant differences in the bacterial contamination rates were observed for the four supermarket chains. This study revealed that retail raw meats are often contaminated with food-borne pathogens; however, there are marked differences in the prevalence of such pathogens in different meats. Raw retail meats are potential vehicles for transmitting food-borne diseases, and our findings stress the need for increased implementation of hazard analysis of critical control point (HACCP) and consumer food safety education efforts.     

Can the high levels of human verocytotoxigenic Escherichia coli O157 infection in rural areas of NE Scotland be explained by consumption of contaminated meat?

AIMS: To determine if contamination levels of Escherichia coli O157 and generic E. coli in retail-minced meat products are greater in rural shops compared with urban shops in Grampian, NE Scotland. We also investigated whether meat from supermarkets and meat from local butcher shops had a similar bacteriological quality. METHODS AND RESULTS: Minced beef and minced lamb were tested from November 2004 to August 2006. Escheichia coli O157 was found at low levels in four samples out of 530 tested samples (0.75%). Generic E. coli were present in 11% of the samples tested, of which 67% came from supermarkets. We observed no significant difference in the prevalence of generic E. coli between rural and urban areas. CONCLUSIONS: Low levels of contamination with E. coli O157 and generic E. coli in retail meat suggest that meat is not a major route of infection in NE Scotland. SIGNIFICANCE AND IMPACT OF THE STUDY: The study does not suggest that the high incidence of E. coli O157 human infection in the rural areas of Grampian is because of meat consumption--this provides further evidence of contact with animals or water being the routes of infection. Hence, risk mitigation should be focussed more on environmental pathways of infection.     

分子标记及其在核桃遗传多样性研究中的应用

核桃是我国重要的坚果和木本油料树种之一,具有重要的学术研究和经济价值。现代分子标记技术的迅速发展为核桃的种质鉴定、遗传育种、遗传多样性分析、亲缘鉴定等提供了崭新途径。本文主要介绍RFLP、RAPD、AFLP及SSR等几种分子标记技术的主要原理、特点以及在核桃遗传多样性方面的研究进展,分析了分子标记在核桃遗传多样性研究中的主要问题,并对其发展提出了展望。     

Effect of supplementing corn- or barley-based feedlot diets with canola oil on faecal shedding of Escherichia coli O157:H7 by steers

AIMS: This study was conducted to evaluate the effect of supplementing barley- or corn-based diets with canola oil on faecal shedding of Escherichia coli O157:H7 by experimentally inoculated feedlot cattle. METHODS AND RESULTS: Four groups of yearling steers fed on barley- or corn-based feedlot diets containing 0% (BA; CO) or 6% canola oil (BA-O; CO-O) were inoculated with 10(10) CFU of a mixture of four nalidixic acid-resistant strains of E. coli O157:H7. The inoculated strains were tracked in oral (mouth swab) and environmental (water, water bowl interface, feed, faecal pat) samples by enrichment and immunomagnetic separation (IMS) for 12 weeks, and in rectally collected faecal samples for 23 weeks (enumeration by dilution plating for 12 weeks; detection by IMS for a further 11 weeks). Levels of E. coli O157:H7 shed in faecal samples over the course of the enumeration period were similar (P = 0.14) among treatments. Disappearance of the inoculated strains from faeces was more rapid (P = 0.009) with barley than with corn, but shedding levels at the end of the enumeration period were similar (P = 0.21) across grain types. Canola oil supplementation did not affect (P = 0.71) the rate of disappearance of E. coli O157:H7 from faeces. The numbers of steers culture positive for E. coli O157:H7 during the enumeration period were similar (P = 0.57) among treatments. During the 11-week detection period, however, more (P < 0.001) steers were E. coli O157:H7-positive in the BA group (15/64) than in BA-O (two of 64), CO (two of 56), or CO-O (one of 56). The organism was present in two of 48 water samples (both CO-O), one of 48 water bowl swabs (BA-O), four of 48 feed samples (two of 12 BA; two of 12 CO-O), 30 of 48 pen floor faecal pat samples, and 296 of 540 mouth swabs (81/144 BA, 80/144 BA-O, 74/126 CO and 61/126 CO-O). CONCLUSION: Supplementing corn or barley-based diets with canola oil did not affect shedding of E. coli O157:H7 by feedlot cattle. SIGNIFICANCE AND IMPACT OF THE STUDY: High-shedding individuals (i.e. 'super shedders') may be responsible for disseminating E. coli O157:H7 among penmates. Faeces on pen floors appears to be a more significant source of infection than are feed or drinking water.     

Enterobacteriaceae associated with meats and meat handling.

The source of Enterobacteriaceae on meats was shown to be associated with the meat-handling work surfaces in two packing plants studies. A total of 2,343 Enterobacteriaceae were isolated and identified from meat samples and work surfaces at the packing plants and at the retail facilities. Escherichia coli biotype I and Serratia liquefaciens were detected at all stages of meat handling, indicating that they may be present in meats throughout the meat-handling system. Enterobacter agglomerans and S. liquefaciens were the predominant Enterobacteriaceae at the retail level, but they had limited indicator potential for sanitation and hygiene, Klebsiella pneumoniae was a frequent isolate among Enterobacteriaceae from meats and meat-handling surfaces in the packing plants but not at the retail level, indicating that this organism might signal unhygienic handling of meats at the retail level.     

Sensitive detection of Escherichia coli O157:H7 in food and water by immunomagnetic separation and solid-phase laser cytometry

Rapid, direct methods are needed to assess active bacterial populations in water and foods. Our objective was to determine the efficiency of bacterial detection by immunomagnetic separation (IMS) and the compatibility of IMS with cyanoditolyl tetrazolium chloride (CTC) incubation to determine respiratory activity, using the pathogen Escherichia coli O157:H7. Counterstaining with a specific fluorescein-conjugated anti-O157 antibody (FAb) following CTC incubation was used to allow confirmation and visualization of bacteria by epifluorescence microscopy. Broth-grown E. coli O157:H7 was used to inoculate fresh ground beef (<17% fat), sterile 0.1% peptone, or water. Inoculated meat was diluted and homogenized in a stomacher and then incubated with paramagnetic beads coated with anti-O157 specific antibody. After IMS, cells with magnetic beads attached were stained with CTC and then an anti-O157 antibody-fluorescein isothiocyanate conjugate and filtered for microscopic enumeration or solid-phase laser cytometry. Enumeration by laser scanning permitted detection of ca. 10 CFU/g of ground beef or <10 CFU/ml of liquid sample. With inoculated meat, the regression results for log-transformed respiring FAb-positive counts of cells recovered on beads versus sorbitol-negative plate counts in the inoculum were as follows: intercept = 1.06, slope = 0.89, and r2 = 0. 95 (n = 13). The corresponding results for inoculated peptone were as follows: intercept = 0.67, slope = 0.88, and r2 = 0.98 (n = 24). Recovery of target bacteria on beads by the IMS-CTC-FAb method, compared with recovery by sorbitol MacConkey agar plating, yielded greater numbers (beef, 6.0 times; peptone, 3.0 times; water, 2.4 times). Thus, within 5 to 7 h, the IMS-CTC-FAb method detected greater numbers of E. coli O157 cells than were detected by plating. The results show that the IMS-CTC-FAb technique with enumeration by either fluorescence microscopy or solid-phase laser scanning cytometry gave results that compared favorably with plating following IMS.     

Evaluation of the effectiveness of a commercially available defined substrate medium and enumeration system for measuring Escherichia coli numbers in faeces and soil samples

AIMS: To determine if a commercially available defined substrate medium and enumeration system could be utilized as an effective and accurate means of enumerating Escherichia coli in environmental samples containing faeces and soil. METHODS AND RESULTS: The samples tested were either inoculated with laboratory grown E. coli or natural E. coli populations in cow faeces. The number of E. coli recovered from faeces and soil samples using the defined substrate medium and enumeration system and a miniaturized MPN method (using traditional media) was compared by analysing the difference between the two methods in relation to the mean. For four of five groups of samples analysed there was no significant difference in the number of E. coli recovered by the two methods (P > 0.05). In one batch the difference was 0.30 log, which while being statistically significant (P < 0.01) was not considered to be biologically significant. CONCLUSION: The commercially available enumeration system was significantly more precise than the miniaturized MPN method (P < 0.001). SIGNIFICANCE AND IMPACT OF THE STUDY: We conclude that the commercially available defined substrate medium and enumeration system is a suitable method for the measurement of E. coli numbers in faeces and soil samples and should provide advantages of increased precision and a reduction in laboratory analysis time.     

Population differences and learning effects in walnut feeding technique by the Japanese squirrel

Feeding efficiency is an important factor explaining diet selection in seed-eating animals. Behavioral adaptation to process and consume the seed is advantageous to improve their feeding rate. The Japanese walnut (Juglans ailanthifolia), which has a nutritious kernel surrounded by a hard shell, is an important food for the Japanese squirrel (Sciurus lis) in lowland mixed-species forests in Japan. In the present study, population differences in feeding technique were examined. In all squirrels captured from the Takao population (with wild Japanese walnuts in their habitat), the optimal feeding technique for quickly opening the walnut shell was observed. However, only 8% of the squirrels from the Fuji population (without walnuts in their habitat) processed the walnuts by the optimal feeding technique. By a repetitive provision of walnuts for 50 days, 14% of squirrels from the Fuji population acquired the optimal feeding technique. Learning effects did not differ significantly between solitary (without a demonstrator) and social conditions (with a demonstrator). The effects of age on learning the feeding technique were examined using individuals from a zoo population that had been naïve for Japanese walnuts. All five individuals aged 6 months old, and half the individuals aged 1–2 years old, ate walnuts by the optimal feeding technique after a 2-month learning period, but none of the individuals older than 3 years did. Consequently, the walnut feeding techniques of the Japanese squirrel are not entirely genetically fixed behavior and can be improved by learning at a young age.