Molecular epidemiology of Yersinia enterocolitica infections

Yersinia enterocolitica is an important food-borne pathogen that can cause yersiniosis in humans and animals. The epidemiology of Y. enterocolitica infections is complex and remains poorly understood. Most cases of yersiniosis occur sporadically without an apparent source. The main sources of human infection are assumed to be pork and pork products, as pigs are a major reservoir of pathogenic Y. enterocolitica. However, no clear evidence shows that such a transmission route exists. Using PCR, the detection rate of pathogenic Y. enterocolitica in raw pork products is high, which reinforces the assumption that these products are a transmission link between pigs and humans. Several different DNA-based methods have been used to characterize Y. enterocolitica strains. However, the high genetic similarity between strains and the predominating genotypes within the bio- and serotype have limited the benefit of these methods in epidemiological studies. Similar DNA patterns have been obtained among human and pig strains of pathogenic Y. enterocolitica, corroborating the view that pigs are an important source of human yersiniosis. Indistinguishable genotypes have also been found between human strains and dog, cat, sheep and wild rodent strains, indicating that these animals are other possible infection sources for humans.     

Bacterial resistance to antimicrobial peptides

Antimicrobial peptides (AMPs) or host defense peptides (HDPs) are vital components of human innate defense system targeting human‐related bacteria. Many bacteria have various mechanisms interfering with AMP activity, causing resistance to AMPs. Since AMPs are considered as potential novel antimicrobial drugs, understanding the mechanisms of bacterial resistance to direct killing of AMPs is of great significance. In this review, a comparative overview of bacterial strategies for resistance to direct killing of various AMPs is presented. Such strategies include bacterial cell envelope modification, AMP degradation, sequestration, expelling, and capsule.     

Radiation resistance and injury of Yersinia enterocolitica

The D values of Yersinia enterocolitica strains IP134, IP107, and WA, irradiated at 25 degrees C in Trypticase soy broth, ranged from 9.7 to 11.8 krad. When irradiated in ground beef at 25 and -30 degrees C, the D value of strain IP107 was 19.5 and 38.8 krad, respectively. Cells suspended in Trypticase soy broth were more sensitive to storage at -20 degrees C than those mixed in ground beef. The percentages of inactivation and of injury (inability to form colonies in the presence of 3.0% NaCl) of cells stored in ground beef for 10 days at -20 degrees C were 70 and 23%, respectively. Prior irradiation did not alter the cell's sensitivity to storage at -20 degrees C, nor did storage at -20 degrees C alter the cell's resistance to irradiation at 25 degrees C. Added NaCl concentrations of up to 4.0% in Trypticase soy agar (TSA) (which contains 0.5% NaCl) had little effect on colony formation at 36 degrees C of unirradiated Y. enterocolitica. With added 4.0% NaCl, 79% of the cells formed colonies at 36 degrees C; with 5.0% NaCl added, no colonies were formed. Although 2.5% NaCl added to ground beef did not sensitize Y. enterocolitica cells to irradiation, when added to TSA it reduced the number of apparent radiation survivors. Cells uninjured by irradiation formed colonies on TSA when incubated at either 36 or 5 degrees C. More survivors of an exposure to 60 krad were capable of recovery and forming colonies on TSA when incubated at 36 degrees C for 1 day than at 5 degrees C for 14 days. This difference in count was considered a manifestation of injury to certain survivors of irradiation.     

Temperature-regulated efflux pump/potassium antiporter system mediates resistance to cationic antimicrobial peptides in Yersinia

Most bacterial pathogens are resistant to cationic antimicrobial peptides (CAMPs) that are key components of the innate immunity of both vertebrates and invertebrates. In Gram-negative bacteria, the known CAMPs resistance mechanisms involve outer membrane (OM) modifications and specifically those in the lipopolysaccharide (LPS) molecule. Here we report, the characterization of a novel CAMPs resistance mechanism present in Yersinia that is dependent on an efflux pump/potassium antiporter system formed by the RosA and RosB proteins. The RosA/RosB system is activated by a temperature shift to 37 degrees C, but is also induced by the presence of the CAMPs, such as polymyxin B. This is the first report of a CAMPs resistance system that is induced by the presence of CAMPs. It is proposed that the RosA/RosB system protects the bacteria by both acidifying the cytoplasm to prevent the CAMPs action and pumping the CAMPs out of the cell.     

Growth temperature-dependent variation in the bacteriophage-inactivating capacity and antigenicity of Yersinia enterocolitica lipopolysaccharide

Growth temperature affected the structure of Yersinia enterocolitica Ye 3827 lipopolysaccharide (LPS). Although Y. enterocolitica Ye 3827 synthesized smooth LPS when grown at a low temperature (25 degrees C), partial smooth-rough transition occurred when the bacteria were grown at the physiological temperature (37 degrees C). The structural alteration was detected by bacteriophage-inactivation assay and chemical and immunological analyses. LPS prepared from bacteria grown at 25 degrees C inactivated a number of bacteriophages that recognize the O-antigenic polysaccharide portion of LPS, whereas more than 3000 times the amount of LPS from bacteria grown at 37 degrees C was required for the same degree of inactivation. The antigenic determinant(s) responsible for the major reaction between 25 degrees C-LPS and anti-25 degree C-bacteria was located on the O-antigenic polysaccharide portion of LPS, but those responsible for the major reaction between 37 degrees C-LPS and anti-37 degrees C-bacteria were located on the R-core or inner portion of LPS.     

Biological activity of synthetic peptides analogous to heat-stable enterotoxin produced by Yersinia enterocolitica

Abstract 3 peptides were synthesized chemically by following the primary structure of heat-stable enterotoxin (ST) produced by Yersinia enterocolitica . A peptide 1–30, having the whole sequence of 30 amino-acid residues, showed a ST activity similar to that of analogue peptide 15–30 composed of the C-terminal 16 amino acid residues. The c-GMP levels of L cells increased through an interaction with peptide 1–30 but not with peptide 15–30, while membranes isolated from broken L cells responded to both. Peptide 1–11, composed of the N-terminal 11 amino-acid residues, showed no biological activity.     

Septicemia due to Yersinia enterocolitica

Despite the increasing number of reports of Yersinia enterocolitica infection in humans, septicemia with this organism has remained a rare complication. A 73-year-old woman presented with fever, jaundice, hepatomegaly and cellulitis. Microorganisms isolated from both skin lesion and blood were biochemically and serologically identified as Yersinia enterocolitica, biotype 4, serotype 3 and lysotype 9b. High agglutinating titres against this organism were demonstrated in the patient''s serum. Complete recovery followed a course of gentamicin sulfate. A household pet was considered, but not proved, to be the source of this infection.     

Arsenic and cadmium resistance in environmental isolates of Yersinia enterocolitica and Yersinia intermedia

Environmental strains of Yersinia enterocolitica representing biotype 1A lack virulence plasmid (pYV) and are regarded as non-pathogenic. Though these occupy a diverse range of environmental niches, nothing is known about their resistance to heavy metals. The minimal inhibitory concentrations (MICs) of various metal ions, namely Ag+, Cu2+, Zn2+, Cd2+, As5+, and As3+, for strains of Yersinia enterocolitica (biotype 1A) and Yersinia intermedia (biotypes 1, 2, and 4), isolated from sewage effluents or pork, were determined. All isolates were resistant (MICs 2.5-5 mM) to Cd2+. The MICs of arsenic varied with bacterial strain and the chemical species of the arsenic used. For the majority of the strains, however, it was between 5-10 mM of Na2HAsO4.7H2O and NaAsO2, and 0.625-2.5 mM of As2O3. Except for one isolate, MICs of Ag+, Cu2+, and Zn2+ for these strains were in the range of 0.3-0.625 mM.     

细菌抗菌肽耐药机制研究进展

细菌对传统抗生素的耐药程度十分严重,寻找克服耐药性的新型抗菌药物已成为当务之急。抗菌肽(antimicrobial peptides,AMPs)是当下较有前景的抗菌药物之一。虽然通常认为,AMPs优先攻击细胞膜的特点使其不会引起广泛的耐药性,但其对特定靶标的识别能力仍为基因突变和细菌耐药性的产生提供了可能。此外,一些细菌还显示出了抵御宿主AMPs的杀伤作用并与宿主细胞共存的能力,相应的细菌防御机制也使其对治疗性AMPs产生抗性,这种交叉抗性近年来也备受关注。这些耐药现象的发现均对AMPs的开发提出了新挑战。本综述就细菌对AMPs耐药的分子机制进行了研究进展的总结,并且对治疗性AMPs与宿主防御肽交叉抗性的相关机制研究进行了归纳,以期寻求新的对抗耐药性的策略。     

Sensitivity of Yersinia enterocolitica to antibiotics]

Sensitivity of 33 foreign and 10 native strains of Y. enterocolitica to tetracycline, aminoglycosides, penicillins, levomycetin and polymyxin was studied. All the strains proved to be resistant to penicillin, oxacillin and ampicillin: they produced penicillinase. The level of resistance to penicillin did not always correlate with penicillin activity. The ability of the native strains to acquire R-factor in vitro from Coli bacteria was shown.     

Molecular basis of resistance to azole antifungals

The increased incidence of invasive mycoses and the emerging problem of antifungal drug resistance has prompted investigations of the underlying molecular mechanisms, particularly for the azole compounds central to current therapy. The target site for the azoles is the ERG11 gene product, the cytochrome P450 lanosterol 14alpha-demethylase, which is part of the ergosterol biosynthetic pathway. The resulting ergosterol depletion renders fungal cells vulnerable to further membrane damage. Development of azole resistance in fungi may occur through increased levels of the cellular target, upregulation of genes controlling drug efflux, alterations in sterol synthesis and decreased affinity of azoles for the cellular target. Here, we review the adaptative changes in fungi, in particular Candida albicans, in response to inhibitors of ergosterol biosynthesis. The molecular mechanisms of azole resistance might help in devising more effective antifungal therapies.     

Enterotoxin production and thermal resistance of Yersinia enterocolitica in milk.

Three of 36 raw milk isolates of Yersinia enterocolitica produced enterotoxin in milk at 25 degrees C, but not at 4 degrees C. No strain tested could survive pasteurization.     

Molecular genetic analysis of a locus required for resistance to antimicrobial peptides in Salmonella typhimurium.

The innate immunity of vertebrates and invertebrates to microbial infection is mediated in part by small cationic peptides with antimicrobial activity. Successful pathogens have evolved mechanisms to withstand the antibiotic activity of these molecules. We have isolated a set of genes from Salmonella typhimurium which are required for virulence and resistance to the antimicrobial peptides melittin and protamine. Sequence analysis of a 5.7 kb segment from the wild-type plasmid conferring resistance to protamine contained five open reading frames: sapA, sapB, sapC, sapD and sapF, organized in an operon structure and transcribed as a 5.3 kb mRNA. SapD and SapF exhibited similarity with the 'ATP binding cassette' family of transporters including the bacterial Opp and SpoOK, involved in the uptake of oligopeptides; the yeast STE6, necessary for the export of a peptide pheromone; and the mammalian mdr, which mediates resistance to chemotherapeutic agents in cancer cells. SapA showed identity with other periplasmic solute binding proteins involved in peptide transport. The SapABCDF system constitutes a novel transporter for enteric bacteria and the first one harboring a periplasmic component with a role in virulence.