Using Chemoattractants to Lure Bacteria to Contact-Killing Surfaces.

Antimicrobial surfaces with covalently attached biocidal functionalities only kill microbes that come into direct contact with the surfaces (contact-killing surfaces). Herein, the activity of contact-killing surfaces is shown to be enhanced by using gradients in the concentration of soluble chemoattractants (CAs) to attract bacteria to the surfaces. Two natural and nonbiocidal CAs (aspartate and glucose) were used to attract bacteria to model surfaces decorated with quaternary ammonium groups (known to kill bacteria that come into contact with them). These results demonstrate the killing of Escherichia coli and Salmonella typhimurium, two common pathogens, at levels 10- to 20-times greater than that of the native surfaces alone. This approach is general and provides new strategies for the design of active or dynamic contact-killing surfaces with enhanced antimicrobial activities.

Bacterial load and inflammation in fetal tissues is not dependent on IL-17a or IL-22 in 10-14 day pregnant mice infected with Listeria monocytogenes.

In this study, we first assessed the effect of intragastric infection of pregnant mice with Listeria monocytogenes on relative expression of select genes associated with T cell subsets. Relative gene expression was moderately increased in placental tissues for IFNγ, IL-4, IL-17a, IL-22, CD3, and FoxP3. To assess the roles of IL-17a and IL-22 in resistance to listeriosis during pregnancy, we compared the severity of maternal and fetal infection in IL-17a((-/-)), IL-22((-/-)), and IL-17a((-/-))/IL-22((-/-)) mice with that of wild type C57BL/6 mice. Intragastric infection with modest numbers of bacterial cells (10(5) CFU) caused reproducible maternal and fetal infection in all four mouse strains. We recovered greater numbers of CFU from the bloodstream of pregnant IL-22((-/-)) mice than pregnant wild type mice. Otherwise we found no significant difference in bacterial load in maternal or fetal tissues (spleen, liver, fetoplacental units) from pregnant IL-17a((-/-)), IL-22((-/-)), or IL-17a((-/-))/IL-22((-/-)) or wild type mice. Nor did we observe histopathologic differences in severity of inflammation in maternal or fetal tissues from the various groups of mice. Although IL-17a and IL-22 are up-regulated in placental tissue, our study suggests that antibacterial resistance and the host inflammatory response are not dependent on IL-17a or IL-22 during infection of mice with L. monocytogenes at 10-14 days of gestation.

Survival and virulence of Salmonella enterica serovar enteritidis filaments induced by reduced water activity.

Salmonella enterica serovar Enteritidis strain E40 filaments were developed under conditions of a reduced water activity (a(w)) of 0.95 in tryptic soy broth (TSB) or tryptic soy agar (TSA) supplemented with 8% or 7% NaCl, respectively. Filament formation was accompanied by an increase of biomass without an increase in CFU and was affected by incubation temperature and the physical milieu. The greatest amount of filaments was recovered from TSA with 7% NaCl and incubation at 30°C. Within 2 h of transfer to fresh TSB, filaments started to septate into normal-sized cells, resulting in a rapid increase in CFU. S. Enteritidis E40 filaments were not more tolerant of low- or high-temperature stresses than nonfilamented control cells. However, there was greater survival of filaments in 10% bile salts after 24 to 48 h of incubation, during pH 2.0 acid challenge for 10 min, and under desiccation on stainless steel surfaces at 25°C and 75.5% relative humidity for 7 days. S. Enteritidis E40 filaments invaded and multiplied within Caco-2 human intestinal epithelial cells to a similar degree as control cells when a comparable CFU of filaments and control cells was used. S. Enteritidis E40 filaments established a successful infection in mice via intragastric inoculation. The filaments colonized the gastrointestinal tract and disseminated to the spleen and liver at levels comparable to those attained by control cells, even when animals were inoculated with 10- to 100-fold fewer CFU. To our knowledge this is the first demonstration of virulence of stress-induced Salmonella filaments in vitro and in vivo. Formation of filaments by Salmonella in food products and food processing environments is significant to food safety, because detection and quantitation of the pathogen may be compromised. The finding that these filaments are virulent further enhances their potential public health impact.

Purine biosynthesis mutants (purA and purB) of serotype 4b Listeria monocytogenes are severely attenuated for systemic infection in intragastrically inoculated A/J Mice.

In this study, we demonstrate that purA and purB transposon mutants of serotype 4b Listeria monocytogenes were severely impaired in their ability to colonize the gastrointestinal tract and cause systemic infection of the spleen, liver, and gallbladder following intragastric inoculation of A/J mice. The mutant strains were also impaired in their ability to multiply within Caco-2 human intestinal epithelial cells. Neither mutant was affected in resistance to synthetic gastric fluid (pH 4.5). These findings indicate that purine biosynthesis is critical for gastrointestinal virulence of L. monocytogenes serotype 4b in mice.

Pregnancy reduces the genetic resistance of C57BL/6 mice to Listeria monocytogenes infection by intragastric inoculation.

In this study, we compared genetically resistant C57BL/6 and susceptible A/J mice for their resistance to Listeria monocytogenes infection during pregnancy. Intragastric infection with modest numbers of bacterial cells (10(5) CFU) caused reproducible fetal infection and abortion in both mouse strains. Bioluminescence imaging demonstrated dissemination of L. monocytogenes cells from maternal to fetal organs within 3 days of intragastric infection. Although non-pregnant C57BL/6 mice were significantly more resistant to infection than non-pregnant A/J mice, C57BL/6 and A/J mice had similar microbial loads (CFU) in maternal and fetal tissues during pregnancy. Inflammation and necrosis, however, were more severe in A/J mice as evaluated by semi-quantitative histopathology. Although the microbial load in fetal tissues was similar for all fetuses within a single uterus, inflammation and necrosis varied among individual fetuses and placentas. We also noted that the uterus is a target for L. monocytogenes infection in non-pregnant mice.

Comparison of systemic Listeria monocytogenes infection in esophageally inoculated mice anesthetized with isoflurane or pentobarbital.

In previous attempts to produce a murine model of gastrointestinal listeriosis, the authors observed that pentobarbital anesthesia greatly increased disease severity in mice that were esophageally inoculated with the bacterium Listeria monocytogenes. In this study, they sought to evaluate the severity of systemic infection in inoculated mice that were anesthetized with isoflurane, an inhalational agent that is safer than pentobarbital and more commonly used for rodent anesthesia. Compared with pentobarbital anesthesia, isoflurane anesthesia resulted in infections of lesser severity, similar to those observed in unanesthetized mice. A pilot study in which mice were anesthetized with isoflurane for 5, 10 or 20 min suggested that this effect was not related to the duration of anesthesia. These results show that isoflurane anesthesia administered for 5 to 20 min does not potentiate the severity of listeriosis infection in mice esophageally inoculated with L. monocytogenes.

Growth of L. monocytogenes strain F2365 on ready-to-eat turkey meat does not enhance gastrointestinal listeriosis in intragastrically inoculated A/J mice.

There have been significant outbreaks of listeriosis associated with consumption of contaminated ready-to-eat (RTE) turkey meat products. In this study, we investigated whether growth on RTE deli turkey meat sends environmental signals to listerial cells that makes them more virulent in the gastrointestinal tract of mice. L. Listeria monocytogenes strain F2365 grew from a starting inoculum of 10(3) CFU/mL to final numbers of 10(8)-10(9) CFU/mL (within 12 days at 10 degrees C) when inoculated onto sliced processed, or whole muscle, turkey breast, or into emulsified whole turkey breast. We did not observe any difference in the numbers of CFU recovered from the spleens and livers of A/J mice inoculated intragastrically with L. monocytogenes grown on sliced turkey meat, in emulsified turkey meat, or in brain heart infusion broth. These results suggest that growth on RTE sliced deli turkey, or in RTE emulsified deli turkey, does not enhance the ability of L. monocytogenes F2365 to cause gastrointestinal listeriosis in intragastrically challenged A/J mice.

The poloxamer P85 is protective against Listeria monocytogenes invasion.

Listeria monocytogenes remains an important foodborne pathogen, and strategies designed to decrease the susceptibility of selected patient populations to foodborne pathogens are therefore desirable. Our objective was to determine if the poloxamer P85 was protective against L. monocytogenes infection. Caco-2 cells were treated with 0.1% (w/v) P85 and challenged with 10(7) L. monocytogenes EGD for 1 hour. A standard gentamicin protection assay was performed to determine invasion differences between the experimental groups. Effects of P85 on the pathogen were studied by measuring bacterial growth and ATP concentrations. In a murine model of listeriosis, FVB mice were administered 150 mg/kg P85 or vehicle control 45 minutes prior to intragastric inoculation of 10(7) L. monocytogenes. Dissemination of the pathogen from the gastrointestinal tract to the liver and spleen was determined 24 hours after bacterial challenge. Pretreatment of Caco-2 cells with P85 significantly decreased L. monocytogenes invasion compared to controls. Repletion of ATP reversed the protective effects of P85. No changes in bacterial ATP or growth profile were detected in P85-treated bacteria. Administration of P85 to mice prior to infection led to decreased dissemination to the liver and spleen compared to vehicle-treated mice. P85 is protective against L. monocytogenes infection when administered prior to bacterial challenge. Modulation of host ATP levels appears to be crucial for the protective effects of P85.

Resistance of Listeria monocytogenes F2365 cells to synthetic gastric fluid is greater following growth on ready-to-eat deli turkey meat than in brain heart infusion broth.

Ready-to-eat (RTE) deli meats have been categorized as high-risk foods for contraction of foodborne listeriosis. Several recent listeriosis outbreaks have been associated with the consumption of RTE deli turkey meat. In this study, we examined whether the growth of Listeria monocytogenes F2365 on commercially prepared RTE deli turkey meat causes listerial cells to become more resistant to inactivation by synthetic gastric fluid (SGF). Listerial cells grown on turkey meat to late logarithmic-early stationary phase were significantly more resistant to SGF at pH 7.0, 5.0, or 3.5 than listerial cells grown in brain heart infusion (BHI) broth. The pH was lower in the fluid in packages of turkey meat than in BHI broth (6.5 versus 7.5). However, listerial cells grown in BHI broth adjusted to a lower pH (6.0) did not exhibit enhanced resistance to SGF. The lesser resistance to SGF of listerial cells grown in BHI broth may be due, in part, to the presence of glucose (0.2%). This study indicates the environment presented by the growth of L. monocytogenes on deli turkey meat affects its ability to survive conditions it encounters in the gastrointestinal tract.

Acute Fetal Demise with First Trimester Maternal Infection Resulting from Listeria monocytogenes in a Nonhuman Primate Model.

Infection with Listeria monocytogenes during pregnancy is associated with miscarriage, preterm birth, and neonatal complications, including sepsis and meningitis. While the risk of these conditions is thought to be greatest during the third trimester of pregnancy, the determinants of fetoplacental susceptibility to infection, the contribution of gestational age, and the in vivo progression of disease at the maternal-fetal interface are poorly understood. We developed a nonhuman primate model of listeriosis to better understand antecedents of adverse pregnancy outcomes in early pregnancy. Four pregnant cynomolgus macaques (Macaca fascicularis) received a single intragastric inoculation between days 36 and 46 of gestation with 107 CFU of an L. monocytogenes strain isolated from a previous cluster of human listeriosis cases that resulted in adverse pregnancy outcomes. Fecal shedding, maternal bacteremia, and fetal demise were consistently noted within 7 to 13 days. Biopsy specimens of maternal liver, spleen, and lymph node displayed variable inflammation and relatively low bacterial burden. In comparison, we observed greater bacterial burden in the decidua and placenta and the highest burden in fetal tissues. Histopathology indicated vasculitis, fibrinoid necrosis, and thrombosis of the decidual spiral arteries, acute chorioamnionitis and villitis in the placenta, and hematogenous infection of the fetus. Vascular pathology suggests early impact of L. monocytogenes infection on spiral arteries in the decidua, which we hypothesize precipitates subsequent placentitis and fetal demise. These results demonstrate that L. monocytogenes tropism for the maternal reproductive tract results in infection of the decidua, placenta, and the fetus itself during the first trimester of pregnancy.IMPORTANCE Although listeriosis is known to cause significant fetal morbidity and mortality, it is typically recognized in the third trimester of human pregnancy. Its impact on early pregnancy is poorly defined. Here we provide evidence that exposure to L. monocytogenes in the first trimester poses a greater risk of fetal loss than currently appreciated. Similarities in human and nonhuman primate placentation, physiology, and reproductive immunology make this work highly relevant to human pregnancy. We highlight the concept that the maternal immune response that protects the mother from serious disease is unable to protect the fetus, a concept relevant to classic TORCH (toxoplasmosis, other, rubella, cytomegalovirus, and herpes) infections and newly illuminated by current Zika virus outbreaks. Studies with this model, using the well-understood organism L. monocytogenes, will permit precise analysis of host-pathogen interactions at the maternal-fetal interface and have broad significance to both recognized and emerging infections in the setting of pregnancy.

Intragastric inoculation with a cocktail of Listeria monocytogenes strains does not potentiate the severity of infection in A/J mice compared to inoculation with the individual strains comprising the cocktail.

Although multistrain cocktails of Listeria monocytogenes are used in food inoculation experiments, no studies, to our knowledge, have been reported that use these cocktails in an intragastric mouse model. In this study, we used a five-strain L. monocytogenes cocktail consisting of strains Scott A, MFS108, 101M, V7, and 310 and a four-strain L. monocytogenes cocktail containing strains Scott A, EGD, H7738, and F2365. Here, we report that intragastric inoculation of anesthetized mice with approximately 106 CFU of a cocktail of L. monocytogenes strains does not result (P > 0.05) in a more severe infection (on the basis of the CFU of Listeria spp. recovered from the spleen, liver, and blood) than inoculation of mice with similar numbers of the individual strains comprising the cocktail. Nor did we observe any consistent relationship between susceptibility of L. monocytogenes strains to inactivation in synthetic gastric fluid in vitro and virulence in mice.

Effects of suspension in emulsified wiener or incubation in wiener packages on the virulence of Listeria monocytogenes Scott A in intragastrically inoculated A/J mice.

Several outbreaks of listeriosis have been associated with contamination of wieners and other ready-to-eat meat products. In this study, we addressed the question of whether emulsification in, or growth on, wieners triggers a response in the listerial cells that makes them more virulent or protects them against the harsh environment of the gastrointestinal tract in mice. Our results indicate that Listeria monocytogenes Scott A grows poorly, if at all, in one brand of commercially prepared wieners inoculated with 5 x 10(3) to 5 x 10(6) CFU per package and incubated at 15 degrees C. Neither L. monocytogenes Scott A emulsified in a slurry of homogenized wieners nor recovered from wiener package fluid after a 7-day incubation at 15 degrees C were more virulent when inoculated into the stomachs of A/J mice than L. monocytogenes Scott A grown in brain heart infusion broth. These findings suggest that the ability of L. monocytogenes Scott A to cause systemic infection following introduction into the gastrointestinal tract was not improved by incubation with wieners or suspension in a meat matrix.

Viability of Escherichia coli O157:H7 during manufacturing and storage of a fermented, semidry soudjouk-style sausage.

Soudjouk-style batter was inoculated with a five-strain mixture of Escherichia coli O157:H7 at about 7.6 log10 CFU/g in each of two trials. The sticks were fermented and dried at 22 degrees C and 50% relative humidity (RH) for 3 days and then at 9 degrees C and 40% RH for 18 h. After being flattened to about 1.25 cm, the sticks were conditioned at 38 degrees C and 70% RH or at 22 degrees C and 50% RH for about 3 days. After the latter conditioning treatment, sticks either were cooked to an internal temperature of 63 degrees C or received no heat treatment. Final mean pH values after conditioning at 22 degrees C and 50% RH for soudjouk manufactured with a starter culture and dextrose (1.0%) and for soudjouk manufactured without a starter culture were about 4.9 and 6.0, respectively. For soudjouk produced with a starter culture, pathogen numbers were reduced by 4.53 and 0.88 log10 CFU/g after conditioning at 38 degrees C and 70% RH and at 22 degrees C and 50% RH, respectively. For soudjouk produced via natural fermentation, pathogen numbers were reduced by 1.39 and 0.09 log10 CFU/g after conditioning at 38 degrees C and 70% RH and at 22 degrees C and 50% RH, respectively. Cooking reduced pathogen numbers to below the levels detectable by direct plating (<1.0 log10 CFU/g) and by enrichment for soudjouk produced with a starter culture and also reduced pathogen numbers by 6.28 log10 CFU/g for soudjouk produced via natural fermentation. However, cooking also resulted in an unacceptable product. In general, the reduction in pathogen numbers achieved by storage at ambient temperature (25 degrees C) was greater than that achieved by storage at cooler temperatures (4 and 15 degrees C), particularly for soudjouk prepared with a starter culture (for which a final pH value of 4.8 and a 6.4-log10 reduction were obtained after 21 days at 25 degrees C) rather than for that prepared without a starter culture (for which a final pH value of 6.1 and a 2.6-log10 reduction were obtained after 21 days at 25 degrees C). These results indicate that naturally fermented old-country-type sausage may allow the survival of E. coli O157:H7 in the absence of controlled fermentation, postfermentation cooking, and/or an ambient-storage processing step.

Reduction in resident microflora, and experimentally inoculated Salmonella enterica, on spinach leaves treated with vinegar and canola oil.

In this study, we explored the use of vinegar, or vinegar and canola oil as a salad dressing, to reduce bacterial levels on spinach leaves. We found that incubation of spinach leaves with various types of vinegar substantially reduced the predominantly gram-negative microflora. A similar response was observed when spinach leaves were incubated with white vinegar mixed in various proportions with canola oil, as used in salad dressing. We assessed the effects of vinegar, or vinegar and oil, on spinach leaves that had been experimentally inoculated with a cocktail of Salmonella enterica strains. Allowing the mixture to sit at room temperature for at least 20 min resulted in a substantial reduction (up to 2.0 log CFU) in numbers of S. enterica. Vinegar and oil caused a limited reduction in CFU (0.5 log) for spinach leaves experimentally inoculated with a cocktail of Listeria monocytogenes strains. These findings suggest that mixing spinach leaves with vinegar and oil as a salad dressing can reduce the bacterial load associated with the spinach leaves, including Salmonella if it is present.

Validation of pepperoni process for control of Shiga toxin-producing Escherichia coli.

The objective of this study was to compare the survival of non-O157 Shiga toxin-producing Escherichia coli (STEC) with E. coli O157:H7 during pepperoni production. Pepperoni batter was inoculated with 7 log CFU/g of a seven-strain STEC mixture, including strains of serotypes O26, O45, O103, O111, O121, O145, and O157. Sausages were fermented to pH ≤4.8, heated at 53.3°C for 1 h, and dried for up to 20 days. STEC strains were enumerated at designated intervals on sorbitol MacConkey (SMAC) and Rainbow (RA) agars; enrichments were completed in modified EC (mEC) broth and nonselective tryptic soy broth (TSB). When plated on SMAC, total E. coli populations decreased 2.6 to 3.5 log after the 1-h heating step at 53.3°C, and a 4.9- to 5-log reduction was observed after 7 days of drying. RA was more sensitive in recovering survivors; log reductions on it were 1.9 to 2.6, 3.8 to 4.2, and 4.6 to 5.3 at the end of cook, and at day 7 and day 14 of drying, respectively. When numbers were less than the limit of detection by direct plating on days 14 and 20 of drying (representing a 5-log kill), no more than one of three samples in each experiment was positive by enrichment with mEC broth; however, STEC strains were recovered in TSB enrichment. Freezing the 7-day dried sausage for 2 to 3 weeks generated an additional 1- to 1.5-log kill. Confirmation by PCR revealed that O103 and O157 had the greatest survival during pepperoni productions, but all serotypes except O111 and O121 were occasionally recovered during drying. This study suggests that non-O157 STEC s trains have comparable or less ability than E. coli O157 to survive the processing steps involved in the manufacture of pepperoni. Processes suitable for control of E. coli O157 will similarly inactivate the other STEC strains tested in this study.

Surfaces modified with nanometer-thick silver-impregnated polymeric films that kill bacteria but support growth of mammalian cells.

Silver is widely used as a biocidal agent in ointments and wound dressings. However, it has also been associated with tissue toxicity and impaired healing. In vitro characterization has also revealed that typical loadings of silver employed in ointments and dressings (approximately 100 microg/cm(2)) lead to cytotoxicity. In this paper, we report the results of an initial study that sought to determine if localization of carefully controlled loadings of silver nanoparticles within molecularly thin films immobilized on surfaces can lead to antimicrobial activity without inducing cytotoxicity. Polymeric thin films of poly(allylamine hydrochloride) (PAH) and poly(acrylic acid) (PAA) were prepared by layer-by-layer deposition and loaded with approximately 0.4 microg/cm(2) to approximately 23.6 microg/cm(2) of silver nanoparticles. Bacterial killing efficiencies of the silver-loaded films were investigated against Staphylococcus epidermidis, a gram-positive bacterium, and it was determined that as little as approximately 0.4 microg/cm(2) of silver in the polymeric films caused a reduction of 6log(10)CFU/mL (99.9999%) bacteria in suspensions incubated in contact with the films (water-borne assays). Significantly, whereas the antibacterial films containing high loadings of silver were found to be toxic to a murine fibroblast cell line (NIH-3T3), the polymeric films containing approximately 0.4 microg/cm(2) of silver were not toxic and allowed attachment, and growth of the mammalian cells. Thus, the results of this study go beyond prior reports by identifying silver-impregnated, polymeric thin films that are compatible with in vitro mammalian cell culture yet exhibit antibacterial activity. These results support the hypothesis that localization of carefully controlled loadings of silver nanoparticles within molecularly thin polymeric films can lead to antimicrobial activity without cytotoxicity. More broadly, this strategy of modifying surfaces with minimal loadings of bioactive molecules indicates the basis of approaches that may permit management of microbial burden in wound beds without impairment of wound healing.

A P60 mutant of Listeria monocytogenes is impaired in its ability to cause infection in intragastrically inoculated mice.

A spontaneous P60 mutant of Listeria monocytogenes was less able to cause systemic infection in A/J mice, following intragastric inoculation, than the parental wild type strain (SLCC 5764, serotype 1/2a). Significantly fewer CFU were recovered from internal organs (spleen, liver, gall bladder) and from the cecum of mice inoculated intragastrically with the P60 mutant than mice inoculated with wild type L. monocytogenes. The P60 mutant also exhibited a diminished ability to invade and multiply within Caco-2 intestinal epithelial cells. These findings indicate that P60 is required for maximal virulence of L. monocytogenes in the gastrointestinal tract of mice.

The aryl hydrocarbon receptor is required for optimal resistance to Listeria monocytogenes infection in mice.

The aryl hydrocarbon receptor (AhR) is part of a powerful signaling system that is triggered by xenobiotic agents such as polychlorinated hydrocarbons and polycyclic aromatic hydrocarbons. Although activation of the AhR by 2,3,7,8-tetrachlorodibenzo-p-dioxin or certain polycyclic aromatic hydrocarbons can lead to immunosuppression, there is also increasing evidence that the AhR regulates certain normal developmental processes. In this study, we asked whether the AhR plays a role in host resistance using murine listeriosis as an experimental system. Our data clearly demonstrate that AhR null C57BL/6J mice (AhR(-/-)) are more susceptible to listeriosis than AhR heterozygous (AhR(+/-)) littermates when inoculated i.v. with log-phase Listeria monocytogenes. AhR(-/-) mice exhibited greater numbers of CFU of L. monocytogenes in the spleen and liver, and greater histopathological changes in the liver than AhR(+/-) mice. Serum levels of IL-6, MCP-1, IFN-gamma, and TNF-alpha were comparable between L. monocytogenes-infected AhR(-/-) and AhR(+/-) mice. Increased levels of IL-12 and IL-10 were observed in L. monocytogenes-infected AhR(-/-) mice. No significant difference was found between AhR(+/-) and AhR(-/-) macrophages ex vivo with regard to their ability to ingest and inhibit intracellular growth of L. monocytogenes. Intracellular cytokine staining of CD4(+) and CD8(+) splenocytes for IFN-gamma and TNF-alpha revealed comparable T cell-mediated responses in AhR(-/-) and AhR(+/-) mice. Previously infected AhR(-/-) and AhR(+/-) mice both exhibited enhanced resistance to reinfection with L. monocytogenes. These data provide the first evidence that AhR is required for optimal resistance but is not essential for adaptive immune response to L. monocytogenes infection.

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.

A/J mice are susceptible and C57BL/6 mice are resistant to Listeria monocytogenes infection by intragastric inoculation.

Previous studies demonstrated that the innate resistance of mice to Listeria monocytogenes infection by intravenous or intraperitoneal inoculation is regulated principally by the Hc locus on mouse chromosome 2. The A/J and C57BL/6 mouse strains were identified as prototype L. monocytogenes-susceptible and -resistant strains, respectively. In the present study, we compared the relative susceptibilities of A/J and C57BL/6 mice to intragastric (i.g.) inoculation with L. monocytogenes. The results of our study indicate that A/J mice are significantly more susceptible than C57BL/6 mice to an i.g. challenge with L. monocytogenes. This was reflected in the estimated 50% lethal doses for the two strains (10(6) and 10(8) CFU for A/J and C57BL/6 mice, respectively) and a more rapid and severe dissemination of the infection to the spleen and liver in A/J mice than in C57BL/6 mice. Histopathological examination of tissues from the infected mice confirmed the greater severity of disease in A/J mice. Clearance of a primary infection enhanced the resistance of both A/J and C57BL/6 mice to reinfection with L. monocytogenes via the gastrointestinal tract. However, the relative difference in susceptibility between the two strains was evident even after immunization. The A/J mouse holds promise as a model for investigating the pathogenesis of gastrointestinal listeriosis because of its ability to develop systemic infection following challenge with numbers of organisms similar to those recovered from some L. monocytogenes-contaminated food products.