Food-borne Isolates Research Articles

Study on the epidemiological characteristics and molecular typing of Salmonella enterica subsp. enterica serovar Senftenberg in Shanghai

To study the molecular epidemiological characteristics of Salmonella enterica subsp. enterica serovar Senftenberg (Salmonella senftenberg) in Shanghai, from 2006 to 2007. A retrospective analysis in 2006 and 2007 was performed to explore the source of food-borne Salmonella senftenberg. The isolates from diarrhea patients between 2006 and 2007 were identified, including biochemical test, hilA and invA gene phenotyping, drug susceptibility, Riboprinter((R)) (RP) and pulsed-field gel electrophoresis (PFGE). Of the diarrhea patients isolates in the monitoring program on non-typhi Salmonella infection in the year of 2006 in Shanghai, number of patients caused by Salmonella senftenberg ranked the third. The stock of Salmonella senftenberg food-born isolates were derived from swine and beef products between 2003 and 2005. All of the strains from diarrhea patients were susceptible to antibiotics except tetracylina (75.6%). With RP and PFGE molecular typing, the two groups (with hydrogen sulfide and hilA, invA gene or without) could be divided into two different independent clone cluster in genetics. 34 strains of diarrhea were divided into 16 PFGE typing-pattern, and among them 12 strains including type 4 (4 strains), type 5 (1 strains), type 6 (6 strains), type 7 (1 strains) and 13 strains including type 11 (3 strains), type 17 (5 strains), type 23 (5 strains) were two different dominant clone cluster. The epidemic of Salmonella senftenberg within 2006 might have been the result of a long period of case occurrence in Shanghai. This rare outbreak belonged to a cluster of outbreaks caused by two different PFGE clone clusters. Data suggested that the genetic clone of Salmonella senftenberg might have been unstable and the source of contamination were complicated, with the characteristics as the obvious decreasing number of patients, with no food-borne isolates in 2007.

Characterization of Listeria monocytogenes Isolates from Human Listeriosis Cases in Italy

The objective of this study was to characterize by serotyping, pulsed-field gel electrophoresis (PFGE), and PCR amplification of virulence genes and markers of epidemic clones I, II, and III (ECI, ECII, and ECIII) 54 human isolates from apparently sporadic cases of infection occurring in the Lombardy region and in the province of Florence, Tuscany, Italy, in the years 1996 to 2007. Listeria monocytogenes isolates were provided by the clinical microbiology laboratories of the Lombardy region and the "Careggi" Hospital of Florence, Tuscany, Italy. Serotyping, PFGE after digestion with the AscI and ApaI enzymes, and PCR amplification for the inlA, inlC, and inlJ genes and ECI, ECII, and ECIII markers were performed according to procedures described previously. Twenty-five (46.3%) L. monocytogenes isolates were assigned to serotype 1/2a, 23 (42.6%) to serotype 4b, and 6 (11.1%) to serotype 1/2b. Thirty-one AscI pulsotypes were recognized among the 54 human isolates. Eleven molecular subtype clusters, of which eight included indistinguishable pulsotypes and three included closely related pulsotypes, were shared by two to seven isolates. Fifteen isolates exhibited unique AscI pulsotypes. Three groups of clustered isolates and two apparently sporadic isolates generated EC amplicons. All strains tested positive for the inlA, inlC, and inlJ genes. Based on the results of serotyping and molecular typing, there were 11 occasions when L. monocytogenes strains with the same subtype were isolated from more than one listeriosis case. A total of 39 out of 54 isolates (72.2%) were attributed to molecular subtype clusters. The results of the study suggest that routine subtyping of L. monocytogenes strains from human listeriosis cases could allow more-timely detection of outbreaks possibly caused by food-borne isolates from a common source and could lead to control of ongoing food exposure, thus preventing the occurrence of more cases.

Systematic Functional Pandemic Strain–Specific Genes, Three Genomic Islands, Two T3SSs in Foodborne, and ClinicalVibrio parahaemolyticusIsolates in China

Vibrio parahaemolyticus is one of the most important pathogens capable of causing foodborne gastroenteritis in China, Japan, and other countries. Pathogenic V. parahaemolyticus has been known to produce either thermostable direct hemolysin (TDH), TDH-related hemolysin (TRH), or both. The emergence of a new clone in 1995, V. parahaemolyticus O3:K6, has resulted in the first documented pandemic spread of V. parahaemolyticus. In this study, 235 isolates from clinical and food sources were characterized by determining the presence of known virulence factors (tdh, trh), systematic genetic markers (toxRS/new, pandemic group-specific sequence [PGS], orf8) specific for V. parahaemolyticus O3:K6 clone and its clonal derivatives, three important genomic islands (GIs) (VPaI-1, VPaI-5, and VPaI-7), and two type III secretion systems (T3SS1 and T3SS2). Our results showed that all 235 isolates harbored all or part of the T3SS1 genes. All the 103 tdh-positive strains harbored all or part of the VPaI-7 and T3SS2 genes. A total of 91 isolates including six foodborne isolates belonged to a pandemic clone in which eight isolates lacked orf8. All pandemic strains harbored VPaI-1 and VPaI-5 except one O4:K68 strain that lacked VPaI-5 altogether. Twelve clinical pathogenic strains had VPaI-7 and T3SS2 but lacked VPaI-1 and VPaI-5. Thirteen nonpathogenic clinical strains and 119 foodborne strains, including six foodborne pathogenic trh-positive strains, only harbored T3SS1 genes. These results indicated that O3:K6 and its serovariants were the main pandemic clone in China. VPaI-1 and VPaI-5 genes were specifically correlated with pandemic strains while VPaI-7 and T3SS2 were closely associated with tdh-positive strains.

A correlation between the virulence and the adhesion of Listeria monocytogenes to silicon nitride: An atomic force microscopy study

Listeria monocytogenes is a facultative intracellular Gram-positive bacterium that is widely distributed in the environment. Despite being pathogenic at the species level, L. monocytogenes in fact comprises a diversity of strains from pathogenic ones that can result in disease and/or mortality to others that are relatively avirulent. The main goal of the current study was to answer the question on whether enhanced binding or attachment of L. monocytogenes to inert surfaces bears any relationship to pathogenicity in food-borne isolates. To answer this question, the nanoscale adhesion forces of eight L. monocytogenes strains that vary in their pathogenicity levels to a model surface of silicon nitride were quantified using atomic force microscopy. The strains used were the highly pathogenic (EGDe, 874, 1002, ATCC 19115), the intermediate pathogenic (ATCC 19112, ATCC 19118), and the non pathogenic (ATCC 15313 and HCC25). Our results indicate that the average nanoscale adhesion (in nN) and the 50% lethal dose (LD50) of strain virulence quantified in mice are logarithmically correlated according to: (nN) = −0.032 ln (LD50) + 1.040, r 2 = 0.96. Such correlation indicates that nanoscale adhesion could potentially be used as a design criterion to distinguish between virulent and avirulent L. monocytogenes strains. Finally, stronger adhesion of virulent strains to inert surfaces modeled by silicon nitride might be a way for pathogenic strains to survive better in the environment and thus increase their likelihood of infecting animals or humans.

Optical mapping and 454 sequencing of Escherichia coli O157 : H7 isolates linked to the US 2006 spinach-associated outbreak

Optical maps for five representative clinical, food-borne and bovine-derived isolates from the 2006 Escherichia coli O157 : H7 outbreak linked to fresh spinach in the United States showed a common set of 14 distinct chromosomal markers that define the outbreak strain. Partial 454 DNA sequencing was used to characterize the optically mapped chromosomal markers. The markers included insertions, deletions, substitutions and a simple single nucleotide polymorphism creating a BamHI site. The Shiga toxin gene profile of the spinach-associated outbreak isolates (stx1(-) stx2(+) stx2c(+)) correlated with prophage insertions different from those in the prototypical EDL933 and Sakai reference strains (stx1(+) stx2(+) stx2c(-)). The prophage occupying the yehV chromosomal position in the spinach-associated outbreak isolates was similar to the stx1(+) EDL933 cryptic prophage V, but it lacked the stx1 gene. In EDL933, the stx2 genes are within prophage BP933-W at the wrbA chromosomal locus; this locus was unoccupied in the spinach outbreak isolates. Instead, the stx2 genes were found within a chimeric BP933-W-like prophage with a different integrase, inserted at the argW locus in the outbreak isolates. An extra set of Shiga toxin genes, stx2c, was found in the outbreak isolates within a prophage integrated at the sbcB locus. The optical maps of two additional clinical isolates from the outbreak showed a single, different prophage variation in each, suggesting that changes occurred in the source strain during the course of this widespread, multi-state outbreak.

INCIDENCE AND DIARRHEGENIC POTENTIAL OF BACILLUS CEREUS IN PASTEURIZED MILK AND CEREAL PRODUCTS IN THAILAND

ABSTRACTBacillus cereus, bacteria that commonly occur in foods, can potentially cause foodborne illness. Two important factors that contribute to the illness are the number of B. cereus in food and the ability of the organism to produce enterotoxins. This study investigated the number of B. cereus cells in dairy and cereal products in Thailand, using the plate count method and the presence of diarrheal‐enterotoxin genes in the isolates through the polymerase chain reaction (PCR). The genes encoding hemolysin BL (hblA, hblC, hblD), nonhemolytic enterotoxin (nheA, nheB, nheC), cytotoxin K (cytK) and enterotoxin FM (entFM) were the targets of the PCR. B. cereus was found in all pasteurized milk samples and in 37.7% of the cereal product samples, ranging from 50 to 1.7 × 103 cfu/g. PCR results revealed that each gene occurred in more than half of the foodborne isolates tested. A large proportion (96%) of the isolates harbored enterotoxin genes and is considered to be potentially diarrhegenic.PRACTICAL APPLICATIONSSignificant frequency of Bacillus cereus contamination in pasteurized milk and cereal products, and the large proportion of diarrhegenic strains among foodborne B. cereus indicate the high risk of foodborne illness that could be caused by consumption of these foods in Thailand. This suggests that B. cereus should not be disregarded in its significance in disease control and prevention programs. Also, as complete elimination of this organism from pasteurized milk and most of the cereal products through the processing steps is not possible, proper handling and storage of these foods should be strictly applied by the food industry. This is necessary in order to prevent the growth of the organism to levels that can cause foodborne illness. This research is also relevant to other developing countries having similar situations as Thailand, where data concerning the number of B. cereus in foods, frequency of contamination and proportion of enterotoxigenic B. cereus are limited, and where B. cereus gastroenteritis could be underestimated.

Formation of biofilm by strains of Listeria monocytogenes isolated from soft cheese 'wara' and its processing environment

Quantification of biofilm formation by 40 Listeria monocytogenes strains from wara soft cheese and its processing environment was assessed on glass vials surfaces. Attachement to glass surface was quantified using a crystal violet binding assay. All the 40 strains produced biofilms after 48 and 72 h incubation at 37oC. No biofilms were formed at 24 h incubation but biofilm formation increased with incubation period in 20 out of the 40 strains. R2 values obtained were 0.0166 and 0.1193 respectively for biofilm formation between 24 and 48 h and 24 and 72 h incubation periods, respectively (P-values of 0.05*). L. monocytogenes strains isolated from wara cheese were generally sensitive to augmentin, streptomycin, claforan, erythromycin, gentamycin, septrin, tarivid, and rocephine and were highly resistant to nitrofuran, fortum, zinnat, and tetracycline. The enhancement of biofilm formation in L. monocytogenes strains from ‘wara’ in this study suggests a relationship with pathogenicity in foodborne isolates. The transfer of antibiotic resistant L. monocytogenes to human via the food chain is a significant health concern.

Multiple-Locus Sequence Typing and Analysis of Toxin Genes in Bacillus cereus Food-Borne Isolates

In the present study we characterized 47 food-borne isolates of Bacillus cereus using multilocus sequence typing (MLST). Newly determined sequences were combined with sequences available in public data banks in order to produce the largest data set possible. Phylogenetic analysis was performed on a total of 296 strains for which MLST sequence information is available, and three main lineages--I, II, and III--within the B. cereus complex were identified. With few exceptions, all food-borne isolates were in group I. The occurrence of horizontal gene transfer (HGT) among various strains was analyzed by several statistical methods, providing evidence of widespread lateral gene transfer within B. cereus. We also investigated the occurrence of toxin-encoding genes, focusing on their evolutionary history within B. cereus. Several patterns were identified, indicating a pivotal role of HGT in the evolution of toxin-encoding genes. Our results indicate that HGT is an important element in shaping the population structure of the B. cereus complex. The results presented here also provide strong evidence of reticulate evolution within the B. cereus complex.

Production of small, acid-soluble spore proteins inClostridium perfringensnonfoodborne gastrointestinal disease isolates

The molecular basis for the differences in heat resistance between spores of Clostridium perfringens food-borne versus nonfoodborne isolates remains unknown. Since a recent study demonstrated the role of small, acid-soluble spore proteins (SASPs) in heat resistance of spores of food-borne isolates, in the current study, we evaluated the expression of SASP-encoding genes (ssp) and the production of SASPs in nonfoodborne isolates. Our results demonstrated the presence of all three ssp genes in five surveyed nonfoodborne isolates. A beta-glucuronidase assay showed that these ssp genes are expressed specifically during sporulation. Furthermore, nonfoodborne isolate F4969 produced SASPs at a level similar to that of food-borne isolate SM101. Collectively, these results suggest that the difference in the levels of heat resistance between spores of food-borne and the nonfoodborne isolates is not the result of impaired expression of ssp genes and (or) decreased production of SASPs in nonfoodborne isolates.

Subtyping of foodborne and environmental isolates of Escherichia coli by multiplex-PCR, rep-PCR, PFGE, ribotyping and AFLP

A total of 54 isolates were characterized by multiplex-PCR for toxin genes and genotyped using several DNA fingerprinting methods: using repetitive extragenic palindromes (REP) and Box primers (rep-PCR), amplified fragment length polymorphism (AFLP), pulsed-field gel electrophoresis (PFGE) and ribotyping. The known-pathogenic strains tested were from food and clinical samples (34 strains) and included serovars O157:H7, O111:H8, O111:H11, O91:H21 and O55:H7. Two type cultures, Escherichia coli K12 (ATCC 29425) and DUP-101 (ATCC 51739), were included as known non-pathogenic strains and an additional 17 previously unclassified isolates from animal fecal samples. Comparisons of genomic DNA fingerprint patterns using unweighted pair group method with arithmetic averages (UPGMA) cluster analysis of Jaccard similarity indices indicated that all methods tested showed a greater similarity between the E. coli O157:H7 strains than to other isolates. On the basis of these studies, we propose that AFLP, REP-PCR, Box-PCR and ribotyping techniques can all be used for discriminating O157:H7 isolates and are preferred for large-scale screening because of the speed and ease of the methods. The PFGE method is the best to discriminate between subtypes of O157:H7 associated with specific outbreak investigations; however, it is more time consuming and unnecessary if subtyping is not required. There are differences between the dendrograms generated from each method and the relationship between the other strains analyzed. However, the fingerprint profiles of the O157:H7 isolates were virtually identical using REP-PCR and Box-PCR enabling easy distinction of the group. Thus, these typing methods have the potential to aid investigators in identifying the source of an outbreak to prevent or control further spread of E. coli O157:H7.

Enterobacter sakazakii: Infectivity and Enterotoxin Production In Vitro and In Vivo

Enterobacter sakazakii has been implicated as the causal organism in a severe form of neonatal meningitis, with reported mortality rates of 40 to 80%. Dried infant formula has been identified as a potential source of the organism in both outbreaks and sporadic cases. In this study, clinical and foodborne isolates of E. sakazakii were evaluated for enterotoxin production by the suckling mouse assay. In addition, suckling mice were challenged both orally and by intraperitoneal injection. Of 18 E. sakazakii strains evaluated, four were found to test positive for enterotoxin production. All strains of E. sakazakii were lethal to suckling mice at 108 CFU per mouse by intraperitoneal injection, while two strains caused death by the peroral route. In in vitro assays, CHO, Vero, and Y-1 cells demonstrated both cell lysis and rounding when exposed to E. sakazakii strain LA filtrates. This is the first report describing any putative virulence factors of E. sakazakii.

Ability of the Listeria monocytogenes strain Scott A to cause systemic infection in mice infected by the intragastric route.

Listeriosis is an important food-borne disease that causes high rates of morbidity and mortality. For reasons that are not clear, most large outbreaks of human listeriosis involve Listeria monocytogenes serotype 4b. Relatively little is known about the pathogenesis of listeriosis following gastrointestinal exposure to food-borne disease isolates of L. monocytogenes. In the present study, we investigated the pathogenesis of systemic infection by the food-borne isolate Scott A in an intragastric (i.g.) mouse challenge model. We found that the severity of infection with L. monocytogenes Scott A was increased in mice made neutropenic by administration of monoclonal antibody RB6-8C5. This observation was similar to a previous report on a study with the laboratory strain L. monocytogenes EGD. Prior administration of sodium bicarbonate did not enhance the virulence of L. monocytogenes strain Scott A for i.g. inoculated mice. Following i.g. inoculation of mice, two serotype 4b strains of L. monocytogenes (Scott A and 101M) achieved a greater bacterial burden in the spleen and liver and elicited more severe histopathological damage to those organs than did a serotype 1/2a strain (EGD) and a serotype 1/2b stain (CM). Of the four strains tested, only strain CM exhibited poor survival in synthetic gastric fluid in vitro. The other three strains exhibited similar patterns of survival at pHs of greater than 5 and relatively rapid (<30 min) loss of viability at pHs of less than 5.0. Growth of L. monocytogenes Scott A at temperatures of 12.5 to 37 degrees C did not affect its ability to cause systemic infection in i.g. inoculated mice. These observations suggest that the serotype 4b L. monocytogenes strains Scott A and 101M possess one or more virulence determinants that make them better able to cause systemic infection following inoculation via the g.i. tract than do the serotype 1/2 strains EGD and CM.

Antimicrobial Resistance of Food-Related Salmonella Isolates, 1999–2000

Salmonellosis is a major foodborne infection in the United States, and strains of Salmonella that are resistant to a variety of antimicrobial agents have become a major public health concern. To estimate the incidence of antimicrobial-resistant Salmonella in our food supply, the U.S. Food and Drug Administration (FDA) has initiated screening of foodborne isolates for sensitivity to antimicrobial agents, including several antibiotics. Salmonella cultures (n = 502) isolated by FDA laboratories during fiscal year 2000 (1 October 1999 through 30 September 2000) from domestic and imported food products and related samples were tested for susceptibility to each of 12 antimicrobial agents using a disc diffusion assay. Because all isolates were resistant to rifampin (5 or 25 μg), only results with the remaining 11 antimicrobial agents are discussed in this paper. Of the 502 isolates, 247 (49.2%) were resistant to one or more antimicrobial agents, and of these 247 isolates, 170 (68.8%) were resistant to one antimicrobial agent, 33 (13.4%) to two antimicrobial agents, 25 (10.1%) to three antimicrobial agents, 7 (2.8%) to four antimicrobial agents, 8 (3.2%) to five antimicrobial agents, and 2 (0.8%) each to six and seven antimicrobial agents. No isolates were resistant to norfloxacin, whereas only seven were resistant to sulfamethoxazole/trimethoprim, six to trimethoprim, three to gentamicin, and one to ciprofloxacin. These results, for the first time, provide a baseline of data on the incidence of antimicrobial-resistant Salmonella in the U.S. food supply, which should be useful in determining the evolution of antimicrobial resistance in the future.

Antibiotic resistance among Listeria, including Listeria monocytogenes, in retail foods.

In the past eight to 10 years, reports of antibiotic resistance in food-borne isolates in many countries have increased, and this work examined the susceptibility of 1001 food isolates of Listeria species. Susceptibility/resistance to eight antibiotics was determined using the Bauer-Kirby disc diffusion assay, and 10.9% of the isolates examined displayed resistance to one or more antibiotics. Resistance to one or more antibiotics was exhibited in 0.6% of Listeria monocytogenes isolates compared with 19.5% of Listeria innocua isolates. Resistance was not observed in Listeria seeligeri or Listeria welshimeri. Resistance to tetracycline (6.7%) and penicillin (3.7%) was the most frequently observed, and while resistance to one antibiotic was most common (9.1%), isolates resistant to two or more antibiotics (1.8%) were also observed. While resistance to the antibiotics most commonly used to treat human listeriosis was not observed in L. monocytogenes, the presence of such resistance in other Listeria species raises the possibility of future acquisition of resistance by L. monocytogenes. The higher level of resistance in L. innocua compared with L. monocytogenes suggests that a species-related ability to acquire resistance to antibiotics exists.

Pooling of Noncollaborative Multilaboratory Data for Evaluation of the Use of DNA Probe Test Kits in Identifying Listeria monocytogenes Strains

The Accuprobe test kit, a DNA probe culture confirmation test kit for identifying bacterial isolates as Listeria monocytogenes was evaluated with 148 food-borne Listeria isolates. The identities of the isolates were confirmed by conventional tests. A subset of the Listeria isolates was also used to evaluate the Gene-Trak Listeria monocytogenes isolation kit. The data obtained with each kit were pooled with data for that kit from independent published studies. Interlaboratory performances were then estimated statistically, to achieve the main features of orthodox interlaboratory collaborative studies. The large number of strains required in this noncollaborative evaluation method advantageously provided a more representative sampling of the potential isolates that may be found in practice. Exact comparability between the two kit studies was not possible nor is it a feature of orthodox collaborative identification kit studies. However, the kits apparently performed equivalently given the different statistical confidences dictated by the study designs.

Isolation of verotoxigenic Escherichia coli from the tasmanian environment

Growing concerns on the emergence of verotoxin producing Escherichia coli (VTEC) in Australia have focused our attention on the possible sources of VTEC within the island state of Tasmania. An analysis of 156 foodsamples and 194 water samples obtained from various areas revealed evidence of eight possible sources. Six strains, with serotypes Ont:Hnt, O86:H −, O88:H −, O126:H21 and O134:H, were isolated from water samples. Two VTEC of serotypes Ont:H8, O81:H − were isolated from raw meat samples. The waterborne isolates produced verocytotoxin, VT1, while both foodborne isolates were strong producers of VT2. Three VTEC isolates produced haemolysins, only one produced enterohaemolysin (EntHly) and the remaining were reported with alpha-haemolysin (α-Hly) activity. An important feature in the majority of isolates from water was their lack of ability to ferment lactose these isolates are routinely overlooked in public health laboratories.

Multilocus enzyme electrophoresis for characterization of Listeria monocytogenes isolates: results of an international comparative study

Multilocus enzyme electrophoresis (MEE) is a standard technique that is used to elucidate the epidemiology of a variety of bacterial species. Recently, the method has been employed by several laboratories for investigations of clinical and foodborne isolates of Listeria monocytogenes. To assess the sensitivity and reproducibility of MEE in characterising L. monocytogenes isolates for epidemiological purposes and, ultimately, to agree on a standard protocol, seven laboratories participated in a blinded study of 80 strains. The strain collection included both epidemiologically related and unrelated isolates. Each laboratory used its own protocol for MEE. The number of enzymes that were assayed by the laboratories ranged from 8 to 23, and the total number of identified electrophoretic types (ETs) varied between 14 and 25. Of the II pairs of duplicate strains, the number of pairs recognised as identical by the seven laboratories ranged from 3 to 10 (median = 8). From 10 to 18 (median = 15) of the 22 groups of epidemiological related strains were recognised as homogeneous by the different laboratories. The discriminatory power of the method, calculated using Simpson's index of diversity for 69 strains (80 strains minus the 11 duplicates), ranged from 0.827 to 0.925. This relatively low discriminatory power is a consequence of a somewhat low genetic diversity of L. monocytogenes compared to other bacterial species. Efforts should be pursued to standardise the method in order to improve the intra- and inter-laboratory reproducibility.

High Prevalence of Multiple Resistance to Antibiotics in 144 Listeria Isolates from Spanish Dairy and Meat Products

The resistance to 12 commonly used antimicrobial agents of 144 foodborne isolates belonging to the genus Listeria (23 L. monocytogenes, 54 L. innocua, 66 L. seeligeri, and 1 L. welshimeri) was tested by using the agar disc-diffusion assay. The Listeria strains were isolated from dairy products (different varieties of unripened fresh and bacteria-ripened hard cheeses made from ewe's, cow's, and goat's milk) and meat products (longaniza, a type of pork sausage). A total of 84 (93%) and 54 (100%) Listeria strains isolated from cheese and pork sausage, respectively, were resistant to multiple antimicrobial agents. More than 80% of the Listeria strains of both food origins were found to be susceptible to penicillin G and ampicillin, whereas the proportion of isolates resistant to the cephalosporins cefotaxime and cefoxitin was nearly 100%. The prevalence of resistance was much higher for isolates from pork sausage (73.8% on average) than for isolates from cheese (20.9%). This marked difference was statistically significant (P < 0.05; chi-square test) for all antibiotics except ampicillin, cefotaxime, and cefoxitin.The strains of the foodborne pathogen L. monocytogenes isolated from cheese were all susceptible to 9 of the 12 antimicrobial agents evaluated. In contrast, more than 80% of the L. monocytogenes strains isolated from pork sausage exhibited resistance to cefotaxime, cefoxitin, tobramycin, amikacin, chloramphenicol, tetracycline, and erythromycin. The appearance of substantial resistance to antibiotics in foodborne Listeria isolates suggests the need for more prudent use of antibiotics by farmers, veterinarians, and physicians.

Studies on the Genes Encoding Botulinum Neurotoxin Type A of Clostridium botulinum from a Variety of Sources

Summary Degenerate oligonucleotide primers, designed to conserved regions of the heavy chain of the botulinum neurotoxin (BoNT), were used to amplify a fragment of approximately 1.1 kb from twenty-five type A Clostridium botulinum strains. These strains were isolated from different geographical locations, from both infant and food-borne botulism. Restriction fragment length polymorphism (RFLP) analysis of the PCR products with Rsa I and Sau3 Al revaled two different groups of BoNT/A genes, designated BoNT/A1 and BoNT/A2. These two groups did not correlate with the origin of the strains, each group containing isolates from infant and food-borne botulism. Organisms isolated from infant botulism in Japan fell into group A2 while all strains examined from the USA, from both infant and food-borne botulism, fell into group A1. Strains from other locations were as follows: food-borne isolates from the UK (groups A1 and A2), Mauritius (group A1) and Venezuela (group A1). In addition, strains encoding BoNT/A1 gave PCR products with primers designated to detect hemagglutinin genes, the products of which form part of the large progenitor toxin complex produced by some strains of C. botulinum , but strains encoding BoNT/A2 gave no such products. In addition, RFLP analysis and probing studies revealed S of the 25 type A strains examined contained unexpressed type B gene sequences.

Genetic basis of tetracycline resistance in food-borne isolates of Listeria innocua.

Eleven of 12 tetracycline-resistant Listeria innocua strains, isolated from chicken or turkey frankfurters and mozzarella cheese, were shown to carry DNA sequences which hybridized with the Tet M probe; of these, two strains also hybridized with Tet K. The remaining strain hybridized with the Tet K probe only. The Tet M determinant appeared to be located on the chromosome; in one case, it was transferable by conjugation to recipients Listeria monocytogenes, Listeria ivanovii, and Enterococcus faecalis.