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PubMed2000

CtsR controls class III heat shock gene expression in the human pathogen Listeria monocytogenes.

Stress proteins play an important role in virulence, yet little is known about the regulation of stress response in pathogens. In the facultative intracellular pathogen Listeria monocytogenes, the Clp ATPases, including ClpC, ClpP and ClpE, are required for stress survival and intracellular growth. The first gene of the clpC operon of L. monocytogenes encodes a homologue of the Bacillus subtilis CtsR repressor of stress response genes. An L. monocytogenes ctsR-deleted mutant displayed enhanced survival under stress conditions (growth in the presence of 2% NaCl or at 42 degrees C), but its level of virulence in the mouse was not affected. The virulence of a wild-type strain constitutively expressing CtsR is significantly attenuated, presumably because of repression of the stress response. Regulation of the L. monocytogenes clpC, clpP and clpE genes was investigated using transcriptional fusions in B. subtilis as a host. The L. monocytogenes ctsR gene was placed under the control of an inducible promoter, and regulation by CtsR and heat shock was demonstrated in vivo in B. subtilis. The purified CtsR protein of L. monocytogenes binds specifically to the clpC, clpP and clpE regulatory regions, and the extent of the CtsR binding sites was defined by DNase I footprinting. Our results demonstrate that this human pathogen possesses a CtsR regulon controlling class III heat shock genes, strikingly similar to that of the saprophyte B. subtilis. This is the first description of a stress response regulatory gene in a pathogen.

Adenosine Triphosphatases↗

PubMed2026

N6-methyladenine DNA modification modulates pathogen virulence in nematodes.

Understanding the global regulatory mechanisms that control pathogen virulence gene expression is essential for elucidating the molecular basis of pathogenicity. N6-methyladenine (6 mA) plays a crucial role in regulating gene expression in response to various environmental stresses; however, its role in pathogen virulence remains largely unexplored. Here, we report the widespread occurrence of 6 mA across 17 nematode isolates and map its genomic landscape in six notorious agriculturally important pathogen root-knot nematodes (RKNs). We demonstrated that 6 mA is characterized by a conserved GAG motif across nematodes, but exhibits species-specific distribution patterns and distinct effects on gene expression. In particular, its enrichment in transposable elements (TEs) differs between polyploid and diploid nematodes, suggesting lineage-specific epigenetic regulation potentially associated with polyploidy. We further identified two functional 6 mA demethylases, MiNMAD-1 and MiNMAD-2, and confirmed their catalytic activity and active sites. Host-induced gene silencing (HIGS) of minmad-1 significantly increased plant resistance to three polyploid RKN species. A detailed functional analysis revealed that minmad-1 knockdown broadly affected gene expression during the parasitic stage, including genes involved in virulence, thereby reducing nematode infectivity. Together, our findings suggest 6 mA demethylase as a key epigenetic regulator of RKNs' virulence, providing new insights into nematode biology and offering promising targets for the development of sustainable control strategies.

Animals↗

PubMed2006

Metabolism of two analgesic agents, tramadol-n-oxide and tramadol, in specific pathogen-free and axenic mice.

The in vivo metabolism of both tramadol-N-oxide (TNO) and tramadol was investigated in urine pools obtained from 0-24 h after a single 300 mg kg-1 oral dose administration of each compound to specific pathogen-free and axenic mice. Unchanged TNO (< or =42% of the initial drug sample), tramadol, and 23 metabolites from TNO-treated mice and unchanged tramadol (< or =15% of the sample) plus 20 metabolites from tramadol-treated mice were profiled, quantified and tentatively identified on the basis of atmospheric pressure ionization mass spectrometry (API-MS) and tandem mass spectrometry (MS/MS) data. Of the tramadol metabolites, five (M1-5) have been previously identified in mice. Of the tramadol and TNO metabolites, six (M18-23) are new metabolites. The tramadol and TNO metabolites were formed via the following seven metabolic pathways: N-oxide reduction (TNO), O/N-demethylation, cyclohexyloxidation, oxidative N-dealkylation, dehydration (TNO), N-oxidation (tramadol), and glucuronidation. Pathways 1-3 appear to be predominant steps forming four major O/N-desmethyl and hydroxycyclohexyl metabolites, and in conjunction with pathway 7, formed six minor glucuronides. Both tramadol-N-oxide and tramadol are extensively metabolized in mice, and no significant qualitative or quantitative differences in metabolism were observed between specific pathogen-free and axenic mice with the exception of a greater amount of unchanged TNO in axenic mice than in specific pathogen-free mice, more M2 in specific pathogen-free mice than in axenic mice in the TNO-dosed mice, and visa versa for M2 of tramadol-dosed mice.

Analgesics, Opioid↗

PubMed2006

Immunization with the biologically active lectin domain of PapGII induces strong adhesion-inhibiting antibody responses but not protection against avian pathogenic Escherichia coli.

The aim of this study was to investigate whether immunization with the sugar binding domain of PapGII (PapGII196) was able to protect chickens against avian pathogenic Escherichia coli. PapGII196 was expressed, purified by Ni-NTA column chromatography and shown to retain its biological activity, as demonstrated by binding to its receptor, globoside. PapGII196 was tested as a vaccine in specific pathogen free broilers and also by vaccinating breeders and assessing protection in their offspring, and using aerosol exposure or air sac inoculation for challenge. Notwithstanding a strong anti-PapGII196 serum IgG response in vaccinated birds in all experiments and inhibition of haemagglutination by serum from PapGII196-vaccinated birds, chickens were not protected against avian pathogenic E. coli. These findings suggest that PapGII may not be a useful candidate for inclusion in vaccines against avian pathogenic E. coli.

Adhesins, Escherichia coli↗

PubMed2006

Rapid transmission of the protozoan parasite Histomonas meleagridis in turkeys and specific pathogen free chickens following cloacal infection with a mono-eukaryotic culture.

In the present investigation, the pathogenicity and transmission of a mono-eukaryotic culture of Histomonas meleagridis for commercial turkeys and specific pathogen free (SPF) chickens is described for the first time. Two separate trials with the same kind of experimental design were performed, one with commercial turkeys and one with SPF chickens. In each experiment, two different groups were included, which were housed in separate rooms. The first group contained four control birds, whereas the second group consisted of 10 infected and four in-contact birds. The birds were infected via the cloaca at 14 days of age with 380,000 cells of a mono-eukaryotic culture of H. meleagridis consisting of a cloned isolate (Turkey/Austria/2922-66/04). Reisolation of the parasite from turkeys and chickens under experimental conditions was performed for the first time. The infected birds started to excrete the parasite as soon as 2 days post infection. Rapid spread of the parasite to in-contact turkeys and chickens was noticed, based on reisolation of live parasites. Reisolation of the pathogen was impossible from two of the four in-contact SPF chickens at any time, whereas all of the infected turkeys were found positive. Intermittent shedding of the parasite was noticed in infected turkeys and SPF chickens, but the phenomenon was much more severe in the SPF chickens as these birds survived the infection. All of the infected and in-contact turkeys died between days 11 and 14 post infection, whereas no death was recorded in the SPF chickens, which were killed 6 weeks after the infection. Typical lesions were recorded in the caeca and livers of the infected turkeys. In addition, a heavy destruction of the bursa of Fabricius was seen in all of the infected and one of the in-contact turkeys. Altogether, the present investigations are of importance for an understanding of the pathogenicity and transmission of H. meleagridis in poultry.

Animals↗

PubMed2000

Pathogen-Driven Outbreaks in Forest Defoliators Revisited: Building Models from Experimental Data.

Models of outbreaks in forest-defoliating insects are typically built from a priori considerations and tested only with long time series of abundances. We instead present a model built from experimental data on the gypsy moth and its nuclear polyhedrosis virus, which has been extensively tested with epidemic data. These data have identified key details of the gypsy moth-virus interaction that are missing from earlier models, including seasonality in host reproduction, delays between host infection and death, and heterogeneity among hosts in their susceptibility to the virus. Allowing for these details produces models in which annual epidemics are followed by bouts of reproduction among surviving hosts and leads to quite different conclusions than earlier models. First, these models suggest that pathogen-driven outbreaks in forest defoliators occur partly because newly hatched insect larvae have higher average susceptibility than do older larvae. Second, the models show that a combination of seasonality and delays between infection and death can lead to unstable cycles in the absence of a stabilizing mechanism; these cycles, however, are stabilized by the levels of heterogeneity in susceptibility that we have observed in our experimental data. Moreover, our experimental estimates of virus transmission rates and levels of heterogeneity in susceptibility in gypsy moth populations give model dynamics that closely approximate the dynamics of real gypsy moth populations. Although we built our models from data for gypsy moth, our models are, nevertheless, quite general. Our conclusions are therefore likely to be true, not just for other defoliator-pathogen interactions, but for many host-pathogen interactions in which seasonality plays an important role. Our models thus give qualitative insight into the dynamics of host-pathogen interactions, while providing a quantitative interpretation of our gypsy moth-virus data.

Lymantria dispar↗

PubMed2005

Evaluation of policies regarding physicians infected with blood-borne pathogens.

OBJECTIVE: Formulating an effective approach to preventing surgeon-to-patient transmission of blood-borne pathogens has been controversial. The objective of our study was to evaluate current community hospital policies, if any, regarding restrictions on surgeons (general surgeons and obstetricians and gynecologists) infected with blood-borne pathogens operating on patients. DESIGN: A survey on hospital policies regarding surgeons infected with blood-borne pathogens was sent to infection control officers at Northern California community hospitals (n = 113). RESULTS: Forty-five hospitals responded to the survey. Of these, only 6 (13.3%) had a policy. Of the 39 (86.7%) that did not have a policy, only 3 hospitals were planning on implementing one. CONCLUSIONS: Many community hospitals are uninterested in instituting a policy regarding the practice of surgeons infected with blood-borne pathogens. Possible reasons include the lack of concern on the individual level, difficulty in defining exposure-prone procedures, and the nature of the relationship between medical staff and community hospitals.

Blood-Borne Pathogens↗

PubMed2005

Protection of healthcare workers from bloodborne pathogens.

PURPOSE OF REVIEW: For decades, healthcare workers have been known to be at risk from acquiring a variety of bloodborne pathogen infections as a result of occupational exposure. Primary prevention of exposures, as recommended by universal precautions guidelines, remains the cornerstone of protecting healthcare workers. Nonetheless, a substantial number of parenteral exposures continue to occur. Updated developments are summarized here, and recommendations for the protection of healthcare workers from bloodborne pathogens are provided. RECENT FINDINGS: The predominant evidence suggests that total percutaneous injuries have decreased over the last decade. Thoughtful adherence to universal precautions remains the primary means of preventing occupational exposures and thus of reducing occupational risk of infection with bloodborne pathogens. A number of studies have provided additional evidence for the efficacy of safety devices in reducing specific subsets of injuries when combined with education and administrative interventions. Barriers to and positive predictors of universal precautions compliance have been identified. Postexposure prophylaxis remains the second line of defense; several authorities have now recommended three antiretroviral agents in this setting. SUMMARY: In summary, almost two decades of experience with universal/standard precautions has resulted in a decrease in parenteral injuries, but much work remains to be done. Vaccines, effective infection control procedures, safer procedures, and safer devices will all be necessary, along with a better understanding of factors that influence healthcare worker behaviors that result in injury. In addition, a number of issues relating to the postexposure management of occupational exposures with bloodborne pathogens need to be better understood.

Anti-Infective Agents↗

PubMed1994

Presence of eaeA sequences in pathogenic and nonpathogenic Escherichia coli strains isolated from weaned rabbits.

Seventy-one Escherichia coli strains isolated from diarrhoeic weaned rabbits from different areas of France were tested for the presence of DNA sequences specific for the EPEC, EHEC, DAEC and EAggEC strains and 16 of them were tested for pathogenicity in animal experiments. High pathogenicity was observed only with strains unable to ferment rhamnose. DNA from all 55 rhamnose-negative O103, O26 and rough strains hybridised with the eaeA probe. Similar hybridisation was obtained with six non-pathogenic rhamnose-positive strains belonging to serogroups O128 and O132. No hybridisation was observed with the other probes. This is the first report of the presence of eaeA sequences in genomic DNA of non-pathogenic strains.

Adhesins, Bacterial↗

PubMed1986

Mucus colonization as a determinant of pathogenicity in intestinal infection by Campylobacter jejuni: a mouse cecal model.

Human isolates of the intestinal pathogen Campylobacter jejuni have been shown to colonize mucus on the outer surface and deep within the intestinal crypts of gnotobiotic or germfree mice. The cecal crypts are preferentially colonized. A model of mucus colonization by C. jejuni in the mouse cecum has been developed, using antibiotic- and magnesium sulfate-treated specific-pathogen-free animals. These spiral-shaped bacteria colonize the mucus in a similar manner to the normal spiral-shaped microbiota. No evidence of adhesion to the intestinal surface was found with a wide variety of microscopic techniques. The campylobacters were seen to be highly motile in living preparations of gut tissue and rapidly tracked along intestinal mucus. Just as many of the normal spiral-shaped bacteria of intestinal surfaces can achieve close association with the epithelium through mucus association and do not adhere to the surface, C. jejuni colonizes the intestinal mucosa via mucus colonization. Thus, a major determinant of pathogenicity in intestinal infection with C. jejuni is proposed to be an ability to colonize intestinal mucus. The possession of specific adhesins is unlikely to be a significant determinant of pathogenicity. Better understanding of the mechanism of mucus association and the properties of the bacterium that are responsible will provide a basis for the rational selection of preventative measures. The model of mucus association in adult antibiotic-treated mice provides an opportunity for colonization studies with variant organisms and immunization studies.

Adhesiveness↗

PubMed2025

Host clustering of Campylobacter species and enteric pathogens in a longitudinal cohort of infants, family members and livestock in rural Eastern Ethiopia.

BACKGROUND: Livestock are recognized as major reservoirs for Campylobacter species and other enteric pathogens, posing infection risks to humans. High prevalence of Campylobacter during early childhood has been linked to environmental enteric dysfunction and stunting, particularly in low-resource settings. METHODS: A total of 280 samples from Campylobacter positive households with complete metadata were analyzed by shotgun metagenomic sequencing followed by bioinformatic analysis via the CZ-ID metagenomic pipeline (Illumina mNGS Pipeline v7.1). Further statistical analyses in JMP PRO 16 explored the microbiome, emphasizing Campylobacter and other enteric pathogens. Two-way hierarchical clustering and split k-mer analysis examined host structuring, patterns of co-infections and genetic relationships. Principal component analysis was used to characterize microbiome composition across the seven sample types. RESULTS: The study identified that microbiome composition was strongly host-driven, with more than 3844 genera detected, and two principal components explaining 62% of the total variation. Twenty-one dominant (based on relative abundance) Campylobacter species showed distinct clustering patterns for humans, ruminants, and broad hosts. The broad-host cluster included the most prevalent species, C. jejuni, C. concisus, and C. coli, present across sample types and a sub-cluster within C. jejuni involving humans, chickens, and ruminants. Campylobacter species from chickens showed strong positive correlations with mothers (r = 0.76), siblings (r = 0.61) and infants (r = 0.54), while co-occurrence analysis found a higher likelihood (Pr > 0.5) of pairs such as C. jejuni with C. coli, C. concisus, and C. showae. Analysis of the top 50 most abundant microbial taxa showed a distinct cluster uniquely present in human stool and absent in all livestock. The study also found frequent co-occurrence of C. jejuni with other enteric pathogens such as Salmonella, and Shigella, particularly in human and chicken. Additionally, instances of Candidatus Campylobacter infans (C. infans) were identified co-occurring with Salmonella and Shigella species in stool samples from infants, mothers, and siblings. CONCLUSIONS: A comprehensive analysis of Campylobacter diversity in humans and livestock in a low-resource setting revealed that infants can be exposed to multiple Campylobacter species early in life. C. jejuni is the dominant species with a propensity for co-occurrence with other notable enteric bacterial pathogens, including Salmonella, and Shigella, especially among infants. Video Abstract.

Animals↗

PubMed1999

Pathogenicity of Sendai viruses adapted into polarized MDCK cells.

Apically and basally released Sendai viruses (SeV) were obtained after infection of polarized Madin-Darby canine kidney (MDCK) cells grown on permeable membrane culture inserts. After 20 passages of adaptation in MDCK cells, we compared their in vivo and in vitro pathogenicity with the parental Mol-strain of SeV. These viruses had comparable in vitro pathogenicity, but the in vivo pathogenicities were varied. The apically released MDCK-adapted virus showed comparable pathogenicity with the parental virus, in contrast with the basally released MDCK-adapted virus, which showed in vivo attenuation.

Animals↗

PubMed2005

Highly pathogenic avian influenza in magpies (Pica pica sericea) in South Korea.

Highly pathogenic avian influenza (HPAI) is an extremely infectious, systemic viral disease of birds that produces high mortality and morbidity. HPAI was diagnosed in the three dead magpies (Pica pica sericea) submitted to the National Veterinary Research and Quarantine Service. At necropsy, the prominent lesions were multifocal or coalescing necrosis of the pancreas with enlargement of the livers and spleens. Microscopically, there were severely necrotizing pancreatitis and lymphocytic meningoencephalitis. Influenza viral antigen was also detected in areas closely associated with histologic lesions. Avian influenza virus was isolated from cecal tonsils and feces of the magpies. The isolated virus was identified as a highly pathogenic H5N1, with hemagglutinin proteolytic cleavage site deduced amino acid sequence of QREKRKKR/GLFGAIAG. To determine the pathogenicity of the isolate, eight 6-wk-old specific-pathogen-free chickens were inoculated intravenously with the virus, and all birds died within 24 hr after inoculation. This is the first report of HPAI in magpies.

Animals↗

PubMedDate not supplied

Sequence analysis of the hemagglutinin gene of H9N2 Korean avian influenza viruses and assessment of the pathogenic potential of isolate MS96.

Sequence analysis of the hemagglutinin (HA) gene of five Korean H9N2 avian influenza virus (AIV) isolates showed that these viruses were closely related and possibly came from the same source. Phylogenetic analysis of the HA1 subunit of H9 subtype isolates revealed that Korean AIV isolates were different from isolates from the poultry markets in Hong Kong in 1997. None of the Korean AIVs had multiple basic amino acids at the HA cleavage site that confer high pathogenicity to some H5 and H7 AIVs. Phylogenetic analysis of the nucleoprotein and matrix gene demonstrated that Korean isolates cluster with Eurasian origin AIVs. The pathogenic potential of one of the isolates (MS96) was assessed after several passages in 14-day-old embryonated chicken eggs (ECE). Fourteen-day-old ECE derivatives of MS96 showed increased HA titer and embryo mortality in eggs; this was apparent after the third passage in 14-day-old ECE. Sequence analysis of the cleavage site of MS96 after the third and tenth passages in 14-day-old ECE revealed no changes in the amino acid sequence. The pathogenicity of MS96 after the tenth passage in 14-day-old eggs (MS96p10(ECE14)) was tested with 4-wk-old specific-pathogen-free chickens. The 14-day-old derivative, MS96p10(ECE14), showed wider tissue tropism and induced more severe clinical signs than the parent virus. Furthermore, after intranasal inoculation of 86-wk-old broiler breeders and 30-wk-old layers, the MS96p10(ECE14) derivative induced more severe signs of depression than the parent virus as well as a transient drop in egg production.

Amino Acid Sequence↗

PubMedDate not supplied

Pathogenicity and immunosuppressive properties of infectious bursal disease "intermediate" strains.

This study was conducted to test the pathogenicity and immunosuppressive effects of seven commercially available infectious bursal disease (IBD) vaccines. These vaccine strains are intermediate in their pathogenicity in susceptible specific-pathogen-free (SPF) chickens. One-day-old and 3-week-old SPF chickens were vaccinated with these vaccines. Two weeks after IBD vaccination, they were vaccinated with Newcastle disease virus (NDV). The pathogenic and immunosuppressive effects of the IBD vaccines were evaluated by the antibody response to NDV vaccination, the bursa: body weight index, and histopathological lesions of the bursa. It was found that these strains were highly variable in their virulence and immunosuppressive properties. Three of the strains tested were found to be highly virulent and immunosuppressive; two others were moderate; and two could be classified as mild.

Animals↗

PubMedDate not supplied

Pathogenicity of Escherichia coli in aerosols for young chickens.

The relative pathogenicity of Esherichia coli isolates from poultry was determined by aerosol exposure of young chickens. Evidence of colisepticemia with airsacculitis and/or pericarditis and perihepatitis was evaluated. A system was devised that included the intratracheal (IT) inoculation of strain SE-17 infectious bronchitis virus (IBV) of chicks at 7 days of age followed by their aerosol exposure to E. coli culture suspensions 2 days later. Each experiment was terminated 6 days later. For comparative purposes in some studies, chicks were housed at 17 C and others at 27 C. The IBV-E. coli challenge procedure proved to be an effective way to determine the relative ability of E. coli isolates to cause death and/or gross lesions in young chickens. With some E. coli isolates, there were minimal or no obvious adverse effects from exposure except when chickens were previously inoculated with IBV. When chicks were housed at 17 C instead of 27 C, slight increases in mortality and decreases in gross lesions were generally observed, probably because the earlier deaths did not allow time for the lesions to become as evident. The E. coli isolate #18344 (Congo Red-positive) was consistently more pathogenic than the Congo Red-negative version of that isolate. Cultures of E. coli previously demonstrated to be pathogenic (VA O1:K1 and DL #29) were among the most pathogenic isolates evaluated in these experiments and were similar to the Congo Red-positive #18344 isolate.(ABSTRACT TRUNCATED AT 250 WORDS)

Aerosols↗

PubMed1997

Management of healthcare workers infected with hepatitis B virus, hepatitis C virus, human immunodeficiency virus, or other bloodborne pathogens. AIDS/TB Committee of the Society for Healthcare Epidemiology of America.

This article provides the current recommendations of the Society for Healthcare Epidemiology of America (SHEA) regarding the management of healthcare workers infected with hepatitis B virus (HBV), hepatitis C virus (HCV), or the human immunodeficiency virus (HIV). For the reasons cited in the article, SHEA now maintains that separate virus-specific management strategies are appropriate for healthcare workers who are infected with these unrelated viruses. SHEA emphasizes the use of appropriate infection control procedures to minimize exposure of patients or providers to blood, emphasizes that transfers of blood from patients to providers and from providers to patients should be avoided, and argues that infected healthcare workers should not be prohibited from participating in patient-care activities solely on the basis of their blood-borne pathogen infection. SHEA recommends that hepatitis B e-antigen-positive healthcare workers routinely should double glove and should not perform those activities that have been identified epidemiologically as associated with a risk for provider-to-patient HBV transmission despite the use of appropriate infection control procedures. SHEA also recommends that HCV- and HIV-infected providers use double gloving for procedures, but recommends that these providers not be excluded from any aspect of patient care unless epidemiologically incriminated in the transmission of these infections despite adequate precautions. SHEA argues for comprehensive education concerning bloodborne pathogens for all healthcare providers and trainees and against mandatory pathogen-specific educational requirements for infected providers. SHEA recommends against specific competence-monitoring procedures directed at these healthcare workers infected with bloodborne pathogens, arguing for managing infected providers in the context of a comprehensive approach to the management of all impaired providers. SHEA emphasizes the importance of worker privacy and medical confidentiality. SHEA emphasizes the importance of offering employees who have disabilities reasonable accommodation for their disabilities. The article discusses exposure management in detail and, in general, recommends adherence to existing guidelines for managing exposures to these agents. Finally, SHEA recommends against routine mandatory testing of providers. Specific details and the rationale for these recommendations are included in the body of the article.

Blood-Borne Pathogens↗

PubMed1984

Eikenella corrodens. An emerging pathogen in head and neck infections.

Eikenella corrodens is a gram-negative, facultative anaerobe that exists as part of the normal oral flora. Its role as a pathogen in human infection has been disputed, but recently its pathogenic potential has been increasingly recognized. A review of the literature reveals the emergence of this organism as a pathogen in human infection. Specific microbiologic characteristics of this organism make it difficult to isolate and evaluate for antibiotic sensitivities. Infections produced by this bacteria are characteristically indolent in nature and are usually associated with oral contamination. Appropriate antibiotic therapy utilizes ampicillin or penicillin. Tetracycline is the drug of choice in the penicillin-allergic patient. Clindamycin resistance is a universal feature. A greater awareness of the pathogenic potential of E corrodens is essential for appropriate recognition and treatment.

Administration, Oral↗