DOCSLIB.ORG

Implementation and Assessment of the Use of Real-Time PCR in Routine Diagnosis for Bordetella Pertussis Detection in Brazil

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Implementation and Assessment of the Use of Real-Time PCR in Routine Diagnosis for Bordetella Pertussis Detection in Brazil Arch Pediatr Infect Dis. 2014 April; 2(2): 196-202. DOI: 10.5812/pedinfect.12505 Research Article Published Online 2014 April 10. Implementation and Assessment of the Use of Real-Time PCR in Routine Diagnosis for Bordetella pertussis Detection in Brazil 1, * 1 1 Daniela Leite , Roberta Morozetti Blanco , Leyva Cecilia Vieira de Melo , Cleiton Eduardo 1 1 1 1 Fiorio , Luciano Moura Martins , Tania Mara Ibelli Vaz , Sueli Aparecida Fernandes , 2 Claudio Tavares Sacchi 1 Center of Bacteriology, National Reference Laboratory for Pertussis, Instituto Adolfo Lutz, Sao Paulo, Brazil 2 Center of Imunology, Instituto Adolfo Lutz, Sao Paulo, Brazil *Corresponding author: Daniela Leite, Center of Bacteriology, National Reference Laboratory for Pertussis, Instituto Adolfo Lutz. Av. Dr. Arnaldo, 351 - 9ºandar, CEP: 01246-902, Sao Paulo-SP, Brazil. Tel: +55-1130682896, Fax: +55-1130819161, E-mail: [email protected]. Received: ; Revised: ; Accepted: May 27, 2013 June 18, 2013 August 27, 2013 Background: Bordetella pertussis is the causative agent of pertussis. In Brazil, laboratory diagnosis of pertussis is based on the culture. In 2010, was standardized the Real-Time PCR TaqMan® in routine diagnosis. Objectives: The aim of this study was to evaluate the impact achieved with the introduction of RT-PCR for the routine diagnosis of pertussis and to compare with the results obtained from culture. Patients and Methods: 4,697 samples of nasopharyngeal secretions collected from suspected pertussis cases and/or contacts were analyzed for RT-PCR and culture, from January 2008 until the end of December 2011. Results: According to the results obtained from the samples 6.9% were culture/RT-PCR positive, 14.8% were positive only for RT-PCR and 0.2% only for culture. Negative samples for both techniques was 3,622 (77.1%) and 1.0% were inconclusive for RT-PCR. Conclusions: The implementation of RT-PCR in routine diagnosis resulted in an increase in laboratory confirmation by almost three times. The RT-PCR assay does not intend to replace the culture technique, but to promote an improvement in the diagnosis of pertussis. Keywords: Bordetella pertussis; Whooping Cough; Diagnosis; Real-Time Polymerase Chain Reaction 1. Background duration of protection following the basic vaccination schedule with a booster dose of wP vaccine or natural Bordetella pertussis, a Gram negative bacterium, is the infection is the same, is estimated to be about 6-12 years. causative agent of whooping cough or pertussis, an in- Some studies show that the duration of protection with fectious respiratory disease of worldwide occurrence the use of aP vaccine is situated within the same time and high prevalence among newborns and children with incomplete immunization. Currently there are eight spe- range (7). cies described in the genus Bordetella including B. pertus- In Brazil, a vaccination program recommended by Min- sis, B. parapertussis, B. holmesii and B. bronchiseptica which istry of Health is three doses with tetravalent vaccine: are the species most commonly associated with respira- diphtheria, wP and tetanus (DTP) + Hib (Haemophilus in- tory infections in humans, although the last one rarely fluenza B) at ages 2, 4 and 6 months age and two boosters infects healthy humans (1-4). of DTP, the first at 15-18 months and the second between Despite mass vaccination programs and good cover- ages 4 and 6 years (8). age in many countries, B. pertussis continues to circulate From 1980 to 1983 more than 40,000 cases of pertussis worldwide and periodically cause epidemics every 3-5 (incidence rate > 30/100,000) were reported in Brazil, but years. Currently, a large number of cases have been re- cases decreased substantially after 1983. In 1990, 15,329 ported among adolescents and adults, to be important cases (incidence rate 10.64/100,000) were reported, the sources of infection for infants and children (5, 6). highest rate observed in the decade. In 1995, 3,798 cases High levels of efficiency have been obtained from both (incidence rate 2.44/100,000) were reported and, there- whole-cell (wP) or acellular (aP) pertussis vaccines. The after, the annual number of cases did not exceed 2,000 Implication for health policy/practice/research/medical education: This paper evaluates the impact achieved with the introduction of RT-PCR for the routine diagnosis of pertussis in Brazil and compares with the results obtained from culture, the gold standard method for diagnosis. Copyright © 2014, Pediartric Infections Research Center. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is prop- erly cited. Leite D et al. (incidence rate 1/100,000). The years 2008 and 2011 re- 3.2. Clinical Samples and Culture ported 1,427 and 2,257 cases (incidence rate 0.71/100,000 We analyzed 4,697 samples of nasopharyngeal secretions and 1.2/100,000) respectively, being considered epidem- collected from patients and/or contacts with suspected ics years by epidemiological surveillance system of the pertussis, assisted in the basic health units distributed in country (9). all Sao Paulo state, from the beginning of January 2008 Estimates from the World Health Organization (WHO) until the end of December 2011. Patients and/or contacts suggest that in 2008, about 16 million cases of pertussis were separated into six age-groups: Group 1: ≤ 2 months occurred worldwide, of which 95% were in developing (n = 1,341); Group 2: 3-6 months (n = 861); Group 3: 7 months countries, where 195,000 children died (10). - 1 year (n = 368); Group 4: 2-9 years (n = 453); Group 5: 10-19 The culture is considered the "gold standard" for detec- years (n = 409) and Group 6: ≥ 20 years (n = 1,034). The age tion of B. pertussis, it is a highly specific method, but with group ranged from eight days old to 85 years old and in 231 a variable sensitivity and lengthy incubation periods. patients and/or contacts was not known. When the material collection is performed at symptom Were considered suspected of pertussis and reported onset (3-4 weeks), high rates of isolation of B. pertussis is to the surveillance system disease: any person who, achieved, but the positivity rate of culture decreases to without other apparent cause, regardless of vaccina- about 15-20% with nasopharyngeal secretion collected six tion status or age, presented symptoms of infection of weeks after the onset of symptoms (11-13). the upper airways with cough lasting at least two weeks Several studies have shown the usefulness of real time and with at least one of the following symptoms: (i) par- polymerase chain reaction (RT-PCR) to detect pathogens. oxysms of coughing, (ii) inspiratory whooping and (iii) The high sensitivity (70-90%) and specificity (86-100%), post-tussive vomiting (i.e. vomiting immediately after along with a shorter response time to results, low risk of coughing) (18). contamination and ease of performance, makes RT-PCR Clinical samples were collected with ultrathin and flex- an alternative method for the diagnosis of many diseas- ible sterile swabs, and immediately placed in a tube with es, among them, the pertussis (13-15). transport Regan-Lowe (RL) medium containing charcoal Traditionally, in Brazil, pertussis laboratory diagnosis agar (Oxoid) semi-solid with 10% sterile defibrinated has been based solely on culture method. Due to the in- sheep blood, and Cephalexin (40 μg/mL). The center of crease in number of notifications of pertussis observed bacteriology, Instituto Adolfo Lutz center and the twelve in other countries (16, 17) and also in Brazil, for over a de- centers of regional laboratories of Instituto Adolfo Lutz cade, there was a need to implement a diagnostic meth- (CLRs - IAL) received and processed the swabs collected at od with higher sensitivity and specificity, with a shorter basic health units according to their areas of coverage. time to release results. Then, in 2010, we standardized the The specimens were cultured on the collection day on Real-Time PCR TaqMan® (RT-PCR) in routine diagnosis of charcoal agar plates (Oxoid) with sheep blood and Ceph- Instituto Adolfo Lutz, the national reference center for alexin incubated at 35-37° C under high humidity ambi- pertussis in Brazil. ent air for twelve days. The samples that showed growth from the third or fourth day, with colonies suggestive 2. Objectives of belonging to the genus Bordetella were confirmed by The aim of this study was to evaluate the impact Gram staining. The biochemical and antigenic character- achieved with the introduction of such technology for izations were realized as described previously (19). the routine diagnosis of pertussis in our settings and to After the culture procedure, the swabs were transferred compare with the results obtained from culture. to dried sterile tubes and kept at -20° C for subsequent phase of extraction, purification and concentration of 3. Patients and Methods bacterial DNA. The swabs processed for culture in CLRs were sent to IAL Central for RT-PCR performing. 3.1. Microorganisms Strains 3.3. Extraction and Purification of Bacterial DNA To evaluate the sensitivity and specificity of RT-PCR, 106 The swabs were thawed, removed from the dry tube B. pertussis strains and a total of 293 strains of microorgan- and stirred with rotational movements into a 1.5 mL mi- isms were analyzed, to several genera and species, which cro tube with 200 µL of free DNA ultrapure sterile water may be present in the human nasopharynx (Table 1). (Roche Diagnostics, Indianapolis, Ind.). From this step, The bacterial strains used in this study belong to the col- the extraction was carried out using QIAampDNA Mini lection of bacteriology research center, Instituto Adolfo Kit (QIAGEN, Valencia, CA) according to the protocol de- Lutz (IAL).
Load more

Recommended publications
  • Official Nh Dhhs Health Alert
    THIS IS AN OFFICIAL NH DHHS HEALTH ALERT Distributed by the NH Health Alert Network [email protected] May 18, 2018, 1300 EDT (1:00 PM EDT) NH-HAN 20180518 Tickborne Diseases in New Hampshire Key Points and Recommendations: 1. Blacklegged ticks transmit at least five different infections in New Hampshire (NH): Lyme disease, Anaplasma, Babesia, Powassan virus, and Borrelia miyamotoi. 2. NH has one of the highest rates of Lyme disease in the nation, and 50-60% of blacklegged ticks sampled from across NH have been found to be infected with Borrelia burgdorferi, the bacterium that causes Lyme disease. 3. NH has experienced a significant increase in human cases of anaplasmosis, with cases more than doubling from 2016 to 2017. The reason for the increase is unknown at this time. 4. The number of new cases of babesiosis also increased in 2017; because Babesia can be transmitted through blood transfusions in addition to tick bites, providers should ask patients with suspected babesiosis whether they have donated blood or received a blood transfusion. 5. Powassan is a newer tickborne disease which has been identified in three NH residents during past seasons in 2013, 2016 and 2017. While uncommon, Powassan can cause a debilitating neurological illness, so providers should maintain an index of suspicion for patients presenting with an unexplained meningoencephalitis. 6. Borrelia miyamotoi infection usually presents with a nonspecific febrile illness similar to other tickborne diseases like anaplasmosis, and has recently been identified in one NH resident. Tests for Lyme disease do not reliably detect Borrelia miyamotoi, so providers should consider specific testing for Borrelia miyamotoi (see Attachment 1) and other pathogens if testing for Lyme disease is negative but a tickborne disease is still suspected.
    [Show full text]
  • Iron Is an Essential Nutrient 4 Bacteria and Humans
    Abstract The bhuTUV and bhuO genes play vital roles in the ability of Brucella abortus to use heme as an iron source and are regulated in an iron-responsive manner by RirA and Irr by Jenifer F. Ojeda April, 2012 Dissertation Advisor: RM Roop II Department of Microbiology and Immunology Brucella abortus is a Gram negative intracellular pathogen that causes the zoonotic disease brucellosis. Antibiotic treatment for brucellosis in humans is prolonged and sometimes followed by relapses. Currently, the United States employs prevention of the illness in humans through cattle vaccinations, eliminating the bacterium in its natural host. Unfortunately, these vaccine strains cause the disease in humans, and Brucella research ultimately aims to identify new vaccine targets as well as alternative treatment options. Brucella abortus resides in the phagosomal compartment of the host macrophage where essential nutrients such as iron are limited. Most bacteria need iron, and within the macrophage, heme is a likely source of iron due to the breakdown of red blood cells by the host macrophage. Heme transporters in Gram negative bacteria are highly conserved, and include components for outer membrane, periplasmic, and cytoplasmic membrane transport. BhuA has been previously characterized as the outer membrane heme transporter of Brucella abortus and here we report that BhuT, BhuU, and BhuV (BhuTUV) are the periplasmic and cytoplasmic heme transport components and that they are required in order for Brucella abortus to transport heme as an iron source. Utilization of heme as an iron source requires the breakdown of heme into ferrous iron, carbon monoxide, and biliverdin by a heme oxygenase.
    [Show full text]
  • BD-CS-057, REV 0 | AUGUST 2017 | Page 1
    EXPLIFY RESPIRATORY PATHOGENS BY NEXT GENERATION SEQUENCING Limitations Negative results do not rule out viral, bacterial, or fungal infections. Targeted, PCR-based tests are generally more sensitive and are preferred when specific pathogens are suspected, especially for DNA viruses (Adenovirus, CMV, HHV6, HSV, and VZV), mycobacteria, and fungi. The analytical sensitivity of this test depends on the cellularity of the sample and the concentration of all microbes present. Analytical sensitivity is assessed using Internal Controls that are added to each sample. Sequencing data for Internal Controls is quantified. Samples with Internal Control values below the validated minimum may have reduced analytical sensitivity or contain inhibitors and are reported as ‘Reduced Analytical Sensitivity’. Additional respiratory pathogens to those reported cannot be excluded in samples with ‘Reduced Analytical Sensitivity’. Due to the complexity of next generation sequencing methodologies, there may be a risk of false-positive results. Contamination with organisms from the upper respiratory tract during specimen collection can also occur. The detection of viral, bacterial, and fungal nucleic acid does not imply organisms causing invasive infection. Results from this test need to be interpreted in conjunction with the clinical history, results of other laboratory tests, epidemiologic information, and other available data. Confirmation of positive results by an alternate method may be indicated in select cases. Validated Organisms BACTERIA Achromobacter
    [Show full text]
  • Phenotypic and Genomic Analyses of Burkholderia Stabilis Clinical Contamination, Switzerland Helena M.B
    RESEARCH Phenotypic and Genomic Analyses of Burkholderia stabilis Clinical Contamination, Switzerland Helena M.B. Seth-Smith, Carlo Casanova, Rami Sommerstein, Dominik M. Meinel,1 Mohamed M.H. Abdelbary,2 Dominique S. Blanc, Sara Droz, Urs Führer, Reto Lienhard, Claudia Lang, Olivier Dubuis, Matthias Schlegel, Andreas Widmer, Peter M. Keller,3 Jonas Marschall, Adrian Egli A recent hospital outbreak related to premoistened gloves pathogens that generally fall within the B. cepacia com- used to wash patients exposed the difficulties of defining plex (Bcc) (1). Burkholderia bacteria have large, flexible, Burkholderia species in clinical settings. The outbreak strain multi-replicon genomes, a large metabolic repertoire, vari- displayed key B. stabilis phenotypes, including the inabil- ous virulence factors, and inherent resistance to many anti- ity to grow at 42°C; we used whole-genome sequencing to microbial drugs (2,3). confirm the pathogen was B. stabilis. The outbreak strain An outbreak of B. stabilis was identified among hos- genome comprises 3 chromosomes and a plasmid, shar- ing an average nucleotide identity of 98.4% with B. stabilis pitalized patients across several cantons in Switzerland ATCC27515 BAA-67, but with 13% novel coding sequenc- during 2015–2016 (4). The bacterium caused bloodstream es. The genome lacks identifiable virulence factors and has infections, noninvasive infections, and wound contamina- no apparent increase in encoded antimicrobial drug resis- tions. The source of the infection was traced to contaminat- tance, few insertion sequences, and few pseudogenes, ed commercially available, premoistened washing gloves suggesting this outbreak was an opportunistic infection by used for bedridden patients. After hospitals discontinued an environmental strain not adapted to human pathogenic- use of these gloves, the outbreak resolved.
    [Show full text]
  • Package Inserts
    Individuals using assistive technology may not be able to fully access the information contained in this file. For assistance, please send an e-mail to: [email protected] and include 508 Accommodation and the title of the document in the subject line of your e-mail. HIGHLIGHTS OF PRESCRIBING INFORMATION -----------------------WARNINGS AND PRECAUTIONS------------------------ These highlights do not include all the information needed to use • Carefully consider benefits and risks before administering VAXELIS to VAXELIS safely and effectively. See full prescribing information for persons with a history of: VAXELIS. - fever ≥40.5°C (≥105°F), hypotonic-hyporesponsive episode (HHE) or VAXELISTM (Diphtheria and Tetanus Toxoids and Acellular Pertussis, persistent, inconsolable crying lasting ≥3 hours within 48 hours after a Inactivated Poliovirus, Haemophilus b Conjugate and Hepatitis B previous pertussis-containing vaccine. (5.2) Vaccine) - seizures within 3 days after a previous pertussis-containing vaccine. (5.2) Suspension for Intramuscular Injection Initial U.S. Approval: 2018 • If Guillain-Barré syndrome occurred within 6 weeks of receipt of a prior vaccine containing tetanus toxoid, the risk for Guillain-Barré syndrome ----------------------------RECENT MAJOR CHANGES ------------------------ may be increased following VAXELIS. (5.3) Dosage and Administration (2.2) 09/2020 • Apnea following intramuscular vaccination has been observed in some ----------------------------INDICATIONS AND USAGE--------------------------- infants born prematurely. The decision about when to administer an VAXELIS is a vaccine indicated for active immunization to prevent intramuscular vaccine, including VAXELIS, to an infant born prematurely diphtheria, tetanus, pertussis, poliomyelitis, hepatitis B, and invasive disease should be based on consideration of the individual infant’s medical status due to Haemophilus influenzae type b.
    [Show full text]
  • Insights Into the Pathogenicity of Burkholderia Pseudomallei
    REVIEWS Melioidosis: insights into the pathogenicity of Burkholderia pseudomallei W. Joost Wiersinga*, Tom van der Poll*, Nicholas J. White‡§, Nicholas P. Day‡§ and Sharon J. Peacock‡§ Abstract | Burkholderia pseudomallei is a potential bioterror agent and the causative agent of melioidosis, a severe disease that is endemic in areas of Southeast Asia and Northern Australia. Infection is often associated with bacterial dissemination to distant sites, and there are many possible disease manifestations, with melioidosis septic shock being the most severe. Eradication of the organism following infection is difficult, with a slow fever-clearance time, the need for prolonged antibiotic therapy and a high rate of relapse if therapy is not completed. Mortality from melioidosis septic shock remains high despite appropriate antimicrobial therapy. Prevention of disease and a reduction in mortality and the rate of relapse are priority areas for future research efforts. Studying how the disease is acquired and the host–pathogen interactions involved will underpin these efforts; this review presents an overview of current knowledge in these areas, highlighting key topics for evaluation. Melioidosis is a serious disease caused by the aerobic, rifamycins, colistin and aminoglycosides), but is usually Gram-negative soil-dwelling bacillus Burkholderia pseu- susceptible to amoxicillin-clavulanate, chloramphenicol, domallei and is most common in Southeast Asia and doxycycline, trimethoprim-sulphamethoxazole, ureido- Northern Australia. Melioidosis is responsible for 20% of penicillins, ceftazidime and carbapenems2,4. Treatment all community-acquired septicaemias and 40% of sepsis- is required for 20 weeks and is divided into intravenous related mortality in northeast Thailand. Reported cases are and oral phases2,4. Initial intravenous therapy is given likely to represent ‘the tip of the iceberg’1,2, as confirmation for 10–14 days; ceftazidime or a carbapenem are the of disease depends on bacterial isolation, a technique that drugs of choice.
    [Show full text]
  • Pseudomonas Aeruginosa1
    The Journal of Immunology IL-17 Is a Critical Component of Vaccine-Induced Protection against Lung Infection by Lipopolysaccharide-Heterologous Strains of Pseudomonas aeruginosa1 Gregory P. Priebe,2*†‡ Rebecca L. Walsh,* Terra A. Cederroth,* Akinobu Kamei,* Yamara S. Coutinho-Sledge,* Joanna B. Goldberg,§ and Gerald B. Pier*‡ In a murine model of acute fatal pneumonia, we previously showed that nasal immunization with a live-attenuated aroA deletant of Pseudomonas aeruginosa strain PAO1 elicited LPS serogroup-specific protection, indicating that opsonic Ab to the LPS O Ag was the most important immune effector. Because P. aeruginosa strain PA14 possesses additional virulence factors, we hypoth- esized that a live-attenuated vaccine based on PA14 might elicit a broader array of immune effectors. Thus, an aroA deletant of PA14, denoted PA14⌬aroA, was constructed. PA14⌬aroA-immunized mice were protected against lethal pneumonia caused not only by the parental strain but also by cytotoxic variants of the O Ag-heterologous P. aeruginosa strains PAO1 and PAO6a,d. Remarkably, serum from PA14⌬aroA-immunized mice had very low levels of opsonic activity against strain PAO1 and could not passively transfer protection, suggesting that an antibody-independent mechanism was needed for the observed cross-serogroup protection. Compared with control mice, PA14⌬aroA-immunized mice had more rapid recruitment of neutrophils to the airways early after challenge. T cells isolated from P. aeruginosa ⌬aroA-immunized mice proliferated and produced IL-17 in high quan- tities after coculture with gentamicin-killed P. aeruginosa. Six hours following challenge, PA14⌬aroA-immunized mice had sig- nificantly higher levels of IL-17 in bronchoalveolar lavage fluid compared with unimmunized, Escherichia coli-immunized, or PAO1⌬aroA-immunized mice.
    [Show full text]
  • Identification of Bordetella Spp. in Respiratory Specimens From
    504 Clinical Microbiology and Infection, Volume 14 Number 5, May 2008 isolates of extended-spectrum-beta-lactamase-producing Original Submission: 27 October 2007; Revised Sub- Shigella sonnei. Ann Trop Med Parasitol 2007; 101: 511–517. mission: 5 December 2007; Accepted: 19 December 21. Rice LB. Controlling antibiotic resistance in the ICU: 2007 different bacteria, different strategies. Cleve Clin J Med 2003; 70: 793–800. Clin Microbiol Infect 2008; 14: 504–506 22. Boyd DA, Tyler S, Christianson S et al. Complete nucleo- 10.1111/j.1469-0691.2008.01968.x tide sequence of a 92-kilobase plasmid harbouring the CTX-M-15 extended spectrum b-lactamase involved in an outbreak in long-term-care facilities in Toronto, Canada. Cystic fibrosis (CF) is an autosomal recessive Antimicrob Agents Chemother 2004; 48: 3758–3764. disease, characterised by defective chloride ion channels that result in multi-organ dysfunction, most notably affecting the respiratory tract. The RESEARCH NOTE alteration in the pulmonary environment is asso- ciated with increased susceptibility to bacterial infection. Recent advances in bacterial taxonomy and improved microbial identification systems Identification of Bordetella spp. in have led to an increasing recognition of the respiratory specimens from individuals diversity of bacterial species involved in CF lung with cystic fibrosis infection. Many such species are opportunistic T. Spilker, A. A. Liwienski and J. J. LiPuma human pathogens, some of which are rarely found in other human infections [1]. Processing Department of Pediatrics and Communicable of CF respiratory cultures therefore employs Diseases, University of Michigan Medical selective media and focuses on detection of School, Ann Arbor, MI, USA uncommon human pathogens.
    [Show full text]
  • Helicobacter Spp. — Food- Or Waterborne Pathogens?
    FRI FOOD SAFETY REVIEWS Helicobacter spp. — Food- or Waterborne Pathogens? M. Ellin Doyle Food Research Institute University of Wisconsin–Madison Madison WI 53706 Contents34B Introduction....................................................................................................................................1 Virulence Factors ...........................................................................................................................2 Associated Diseases .......................................................................................................................2 Gastrointestinal Disease .........................................................................................................2 Neurological Disease..............................................................................................................3 Other Diseases........................................................................................................................4 Epidemiology.................................................................................................................................4 Prevalence..............................................................................................................................4 Transmission ..........................................................................................................................4 Summary .......................................................................................................................................5
    [Show full text]
  • Metagenomics Analysis Reveals the Microbial Communities
    diversity Article Metagenomics Analysis Reveals the Microbial Communities, Antimicrobial Resistance Gene Diversity and Potential Pathogen Transmission Risk of Two Different Landfills in China Shan Wan 1,†, Min Xia 2,†, Jie Tao 1, Yanjun Pang 1, Fugen Yu 1,* , Jun Wu 3,* and Shanping Chen 2,* 1 State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China; [email protected] (S.W.); [email protected] (J.T.); [email protected] (Y.P.) 2 Shanghai Environmental Sanitation Engineering Design Institute Co., Ltd., Shanghai 200232, China; [email protected] 3 State Key Laboratory of Pollution Control and Resource Reuse, School of Environment, Nanjing University, Nanjing 210023, China * Correspondence: [email protected] (F.Y.); [email protected] (J.W.); [email protected] (S.C.) † There authors contribute equally to this work. Abstract: In this study, we used a metagenomic approach to analyze microbial communities, antibiotic resistance gene diversity, and human pathogenic bacterium composition in two typical Citation: Wan, S.; Xia, M.; Tao, J.; landfills in China. Results showed that the phyla Proteobacteria, Bacteroidetes, and Actinobacte- Pang, Y.; Yu, F.; Wu, J.; Chen, S. ria were predominant in the two landfills, and archaea and fungi were also detected. The genera Metagenomics Analysis Reveals the Methanoculleus, Lysobacter, and Pseudomonas were predominantly present in all samples. sul2, sul1, Microbial Communities, tetX, and adeF were the four most abundant antibiotic resistance genes. Sixty-nine bacterial pathogens Antimicrobial Resistance Gene were identified from the two landfills, with Klebsiella pneumoniae, Bordetella pertussis, Pseudomonas Diversity and Potential Pathogen aeruginosa, and Bacillus cereus as the major pathogenic microorganisms, indicating the existence of Transmission Risk of Two Different potential environmental risk in landfills.
    [Show full text]
  • Structural and Functional Effects of Bordetella Avium Infection in the Turkey Respiratory Tract William George Van Alstine Iowa State University
    Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1987 Structural and functional effects of Bordetella avium infection in the turkey respiratory tract William George Van Alstine Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Animal Sciences Commons, and the Veterinary Medicine Commons Recommended Citation Van Alstine, William George, "Structural and functional effects of Bordetella avium infection in the turkey respiratory tract " (1987). Retrospective Theses and Dissertations. 11655. https://lib.dr.iastate.edu/rtd/11655 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS While the most advanced technology has been used to photograph and reproduce this manuscript, the quality of the reproduction is heavily dependent upon the quality of the material submitted. For example: • Manuscript pages may have indistinct print. In such cases, the best available copy has been filmed. • Manuscripts may not always be complete. In such cases, a note will indicate that it is not possible to obtain missing pages. • Copyrighted material may have been removed from the manuscript. In such cases, a note will indicate the deletion. Oversize materials (e.g., maps, drawings, and charts) are photographed by sectioning the original, beginning at the upper left-hand comer and continuing from left to right in equal sections with small overlaps.
    [Show full text]
  • Innate and Adaptive Immune Responses Against Bordetella Pertussis and Pseudomonas Aeruginosa in a Murine Model of Mucosal Vaccination Against Respiratory Infection
    Article Innate and Adaptive Immune Responses against Bordetella pertussis and Pseudomonas aeruginosa in a Murine Model of Mucosal Vaccination against Respiratory Infection Catherine B. Blackwood y, Emel Sen-Kilic y , Dylan T. Boehm, Jesse M. Hall, Melinda E. Varney z, Ting Y. Wong, Shelby D. Bradford, Justin R. Bevere, William T. Witt, F. Heath Damron and Mariette Barbier * West Virginia University Vaccine Development Center, Department of Microbiology, Immunology and Cell Biology, 64 Medical Center Drive, Morgantown, WV 26505, USA; [email protected] (C.B.B.); [email protected] (E.S.-K.); [email protected] (D.T.B.); [email protected] (J.M.H.); [email protected] (M.E.V.); [email protected] (T.Y.W.); [email protected] (S.D.B.); [email protected] (J.R.B.); [email protected] (W.T.W.); [email protected] (F.H.D.) * Correspondence: [email protected] The authors contributed equally. y Current affiliation: Marshall University, Department of Pharmaceutical Science and Research, One John z Marshall Drive, Huntington, WV 25755, USA. Received: 28 September 2020; Accepted: 28 October 2020; Published: 3 November 2020 Abstract: Whole cell vaccines are frequently the first generation of vaccines tested for pathogens and can inform the design of subsequent acellular or subunit vaccines. For respiratory pathogens, administration of vaccines at the mucosal surface can facilitate the generation of a localized mucosal immune response. Here, we examined the innate and vaccine-induced immune responses to infection by two respiratory pathogens: Bordetella pertussis and Pseudomonas aeruginosa. In a model of intranasal administration of whole cell vaccines (WCVs) with the adjuvant curdlan, we examined local and systemic immune responses following infection.
    [Show full text]