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1.
PLoS One ; 19(7): e0294853, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38950318

RESUMO

Feral populations of aoudad (Ammotragus lervia) occur in Texas bighorn sheep (Ovis canadensis) habitat and pose several conceptual ecological threats to bighorn sheep re-establishment efforts. The potential threat of disease transmission from aoudad to bighorn sheep may exacerbate these issues, but the host competency of aoudad and subsequent pathophysiology and transmissibility of pneumonic pathogens involved in the bighorn sheep respiratory disease complex is largely unknown. Because the largest population-limiting diseases of bighorn sheep involve pathogens causing bronchopneumonia, we evaluated the host competency of aoudad for Mycoplasma ovipneumoniae and leukotoxigenic Pasteurellaceae. Specifically, we described the shedding dynamics, pathogen carriage, seroconversion, clinical patterns, and pathological effects of experimental infection among wild aoudad held in captivity. We found that aoudad are competent hosts capable of maintaining and intraspecifically transmitting Mycoplasma ovipneumoniae and Pasteurellaceae and can shed the bacteria for 53 days after exposure. Aoudad developed limited clinical signs and pathological findings ranged from mild chronic lymphohistiocytic bronchointerstitial pneumonia to severe and acute suppurative pneumonia, similarly, observed in bighorn sheep infected with Mycoplasma spp. and Pasteurellaceae bacteria, respectively. Furthermore, as expected, clinical signs and lesions were often more severe in aoudad inoculated with a combination of Mycoplasma ovipneumoniae and Pasteurellaceae as compared to aoudad inoculated with only Mycoplasma ovipneumoniae. There may be evidence of interindividual susceptibility, pathogenicity, and/or transmissibility, indicated by individual aoudad maintaining varying severities of chronic infection who may be carriers continuously shedding pathogens. This is the first study to date to demonstrate that aoudad are a conceptual disease transmission threat to sympatric bighorn sheep populations due to their host competency and intraspecific transmission capabilities.


Assuntos
Mycoplasma ovipneumoniae , Pasteurellaceae , Pneumonia por Mycoplasma , Animais , Mycoplasma ovipneumoniae/patogenicidade , Pasteurellaceae/patogenicidade , Pneumonia por Mycoplasma/transmissão , Pneumonia por Mycoplasma/veterinária , Pneumonia por Mycoplasma/microbiologia , Ovinos , Carneiro da Montanha/microbiologia , Ruminantes/microbiologia , Doenças dos Ovinos/transmissão , Doenças dos Ovinos/microbiologia , Infecções por Pasteurellaceae/transmissão , Infecções por Pasteurellaceae/microbiologia , Infecções por Pasteurellaceae/veterinária , Feminino
2.
Vet Microbiol ; 295: 110147, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38885549

RESUMO

In April 2020, two cows in Japan, developed reproductive disorders accompanied by vaginitis with purulent discharge within 3 days of artificial insemination (AI) with the same lot of frozen semen. Histophilus somni was isolated from the vaginal swabs of both cows as well as from the same lot of frozen semen used for the AI. This incident marks the first reported case of H. somni infection in cattle through AI. The major outer membrane protein gene sequences and pulsed-field gel electrophoresis profiles of the isolates were identical. Moreover, we investigated the antimicrobial activity of 12 frozen semen straws against an H. somni isolate using a disk diffusion test. These straws were sourced from five AI centers and included the same lot of semen used for the AI. Although the composition of semen diluents from individual AI centers is not publicly available, both the same lot of frozen semen used in the AI and other lots produced by the same manufacturer showed lower antimicrobial activity than semen from other manufacturers. These results strongly suggest that the two vaginitis were caused by AI using H. somni-contaminated frozen semen because of insufficient antimicrobial activity to inhibit bacterial growth. The minimum inhibitory concentrations of the six antimicrobials recommended for addition to frozen semen in isolates were below the recommended concentrations, suggesting that proper addition could have prevented this incident. This highlights the importance of conducting periodical checks on the antibacterial activity of frozen semen to prevent the transmission of pathogens via AI.


Assuntos
Doenças dos Bovinos , Inseminação Artificial , Pasteurellaceae , Sêmen , Feminino , Inseminação Artificial/veterinária , Animais , Bovinos , Sêmen/microbiologia , Pasteurellaceae/efeitos dos fármacos , Pasteurellaceae/isolamento & purificação , Doenças dos Bovinos/microbiologia , Masculino , Descarga Vaginal/veterinária , Descarga Vaginal/microbiologia , Antibacterianos/farmacologia , Infecções por Pasteurellaceae/veterinária , Infecções por Pasteurellaceae/microbiologia , Vaginite/microbiologia , Vaginite/veterinária , Testes de Sensibilidade Microbiana , Preservação do Sêmen/veterinária , Japão
3.
Eur J Clin Microbiol Infect Dis ; 43(7): 1487-1493, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38780754

RESUMO

Aggregatibacter actinomycetemcomitans (Aa), a Gram-negative coccobacillus commonly associated with endocarditis, poses a rare diagnostic challenge in pediatric cases. The presentation of two pediatric cases-myositis and chest mass-highlights novel aspects, including unusual symptom presentations in children which can be mistaken for malignancy. The limited sensitivity of standard blood tests complicates diagnosis, leading to delayed diagnosis and treatment. Representative samples must be taken, especially if blood cultures are negative. Despite advances in detection methods, diagnosing Aa infection remains difficult due to its rarity in children and variable clinical presentation. In conclusion, a comprehensive understanding of Aa infection in children is essential for early and effective diagnostic and therapeutic management.


Assuntos
Aggregatibacter actinomycetemcomitans , Infecções por Pasteurellaceae , Humanos , Aggregatibacter actinomycetemcomitans/isolamento & purificação , Masculino , Infecções por Pasteurellaceae/diagnóstico , Infecções por Pasteurellaceae/microbiologia , Feminino , Criança , Miosite/microbiologia , Miosite/diagnóstico , Antibacterianos/uso terapêutico , Pré-Escolar
5.
Medicina (B Aires) ; 84(2): 356-358, 2024.
Artigo em Espanhol | MEDLINE | ID: mdl-38683524

RESUMO

Brain abscess is a focal suppurative process produced in most cases by bacterial agents. Aggregatibacter aphrophilus is a gram-negative bacteria belonging to the HACEK group, which causes infective endocarditis, liver abscesses, among others. Brain abscesses secondary to this germ are rare and, in most cases, it is associated with contact with pets, poor dental hygiene or dental procedures. Treatment consists of drainage of the abscess (greater than 2.5 cm) combined with antibiotic therapy, ideally beta-lactams. The case of a 64-year-old male patient with no relevant history is here presented. He was admitted to the emergency service due to headache, hemianopsia of a week's duration and later tonic-clonic seizures, in whom imaging studies and culture of a brain lesion subsequently revealed a brain abscess due to A. aphrophilus. This case aims to illustrate about the rarity of this infection, because A. aphrophilus is a normal part of the oropharyngeal flora and respiratory tract, in which it rarely causes invasive bacteremia.


El absceso cerebral es un proceso supurativo focal producido en la mayoría de los casos por agentes bacterianos. Aggregatibacter aphrophilus es una bacteria gram negativa perteneciente al grupo HACEK, causante de endocarditis infecciosa, abscesos hepáticos, entre otras. Los abscesos cerebrales secundarios a este germen son infrecuentes y en la mayoría de los casos asociados a contactos con animales domésticos, pobre higiene dental o procedimientos odontológicos. El tratamiento consiste en drenaje del absceso (mayores de 2.5 cm) combinado con terapia antibiótica, idealmente betalactámicos. Se presenta el caso de un paciente varón de 64 años sin antecedentes de relevancia quien ingresó al servicio de emergencias por cuadro de cefalea, hemianopsias de una semana de evolución y posteriormente crisis tónico clónicas, en quien posteriormente en estudios imagenológicos y cultivo de lesión cerebral se arribó al diagnóstico de absceso cerebral por A. aphrophilus. Este informe tiene como objetivo ilustrar al lector sobre la rareza de esta infección, debido a que A. aphrophilus forma parte normal de la flora orofaríngea y del tracto respiratorio, en los que rara vez ocasiona bacteriemias invasivas.


Assuntos
Aggregatibacter aphrophilus , Abscesso Encefálico , Infecções por Pasteurellaceae , Abscesso Encefálico/microbiologia , Abscesso Encefálico/etiologia , Abscesso Encefálico/diagnóstico por imagem , Abscesso Encefálico/tratamento farmacológico , Humanos , Masculino , Aggregatibacter aphrophilus/isolamento & purificação , Pessoa de Meia-Idade , Infecções por Pasteurellaceae/microbiologia , Antibacterianos/uso terapêutico , Drenagem
6.
Vet Pathol ; 61(4): 604-608, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38440930

RESUMO

Mannheimia haemolytica-associated abomasitis has been clinically described as a cause of sudden death in lambs, but it is poorly characterized. We describe the pathological features of a severe fibrinonecrotizing abomasitis in 3 lambs that died suddenly. All 3 abomasums had a thickened submucosa due to edema and necrotic areas delimited by bands of degenerate neutrophils with slender nuclei (oat cells) and angiocentric distributions. The overlying mucosa was congested. Myriads of gram-negative coccobacilli were observed within the oat cell bands. M. haemolytica was isolated from the abomasum in all 3 animals and was serotyped as A2 in one of them. Pericarditis and pleuritis were observed in 2 of the lambs. Clostridium spp. were isolated in 1 lamb and detected by immunohistochemistry in the 3 animals, suggesting clostridial co-infection. M. haemolytica should be considered among the differential diagnoses of necrotizing abomasitis in lambs.


Assuntos
Abomaso , Mannheimia haemolytica , Necrose , Infecções por Pasteurellaceae , Doenças dos Ovinos , Animais , Mannheimia haemolytica/isolamento & purificação , Doenças dos Ovinos/patologia , Doenças dos Ovinos/microbiologia , Ovinos , Abomaso/patologia , Abomaso/microbiologia , Infecções por Pasteurellaceae/veterinária , Infecções por Pasteurellaceae/patologia , Infecções por Pasteurellaceae/microbiologia , Necrose/veterinária , Necrose/patologia , Necrose/microbiologia , Gastropatias/veterinária , Gastropatias/patologia , Gastropatias/microbiologia , Masculino , Feminino , Imuno-Histoquímica/veterinária
7.
Trop Anim Health Prod ; 55(6): 383, 2023 Oct 27.
Artigo em Inglês | MEDLINE | ID: mdl-37889324

RESUMO

Gallibacterium anatis (G. anatis), a member of the Pasteurellaceae family, normally inhabits the upper respiratory and lower genital tracts of poultry. However, under certain circumstances of immunosuppression, co-infection (especially with Escherichia coli or Mycoplasma), or various stressors, G. anatis caused respiratory, reproductive, and systemic diseases. Infection with G. anatis has emerged in different countries worldwide. The bacterium affects mainly chickens; however, other species of domestic and wild birds may get infected. Horizontal, vertical, and venereal routes of G. anatis infection have been reported. The pathogenicity of G. anatis is principally related to the presence of some essential virulence factors such as Gallibacterium toxin A, fimbriae, haemagglutinin, outer membrane vesicles, capsule, biofilms, and protease. The clinical picture of G. anatis infection is mainly represented as tracheitis, oophoritis, salpingitis, and peritonitis, while other lesions may be noted in cases of concomitant infection. Control of such infection depends mainly on applying biosecurity measures and vaccination. The antimicrobial sensitivity test is necessary for the correct treatment of G. anatis. However, the development of multiple drug resistance is common. This review article sheds light on G. anatis regarding history, susceptibility, dissemination, virulence factors, pathogenesis, clinical picture, diagnosis, and control measures.


Assuntos
Infecções por Pasteurellaceae , Pasteurellaceae , Doenças das Aves Domésticas , Feminino , Animais , Aves Domésticas , Galinhas , Infecções por Pasteurellaceae/veterinária , Infecções por Pasteurellaceae/microbiologia , Fatores de Virulência , Escherichia coli , Doenças das Aves Domésticas/microbiologia
8.
Microbiologyopen ; 12(3): e1353, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37379423

RESUMO

Glaesserella parasuis, Mycoplasma hyorhinis, and Mycoplasma hyosynoviae are important porcine pathogens responsible for polyserositis, polyarthritis, meningitis, pneumonia, and septicemia causing significant economic losses in the swine industry. A new multiplex quantitative polymerase chain reaction (qPCR) was designed on one hand for the detection of G. parasuis and the virulence marker vtaA to distinguish between highly virulent and non-virulent strains. On the other hand, fluorescent probes were established for the detection and identification of both M. hyorhinis and M. hyosynoviae targeting 16S ribosomal RNA genes. The development of the qPCR was based on reference strains of 15 known serovars of G. parasuis, as well as on the type strains M. hyorhinis ATCC 17981T and M. hyosynoviae NCTC 10167T . The new qPCR was further evaluated using 21 G. parasuis, 26 M. hyorhinis, and 3 M. hyosynoviae field isolates. Moreover, a pilot study including different clinical specimens of 42 diseased pigs was performed. The specificity of the assay was 100% without cross-reactivity or detection of other bacterial swine pathogens. The sensitivity of the new qPCR was demonstrated to be between 11-180 genome equivalents (GE) of DNA for M. hyosynoviae and M. hyorhinis, and 140-1200 GE for G. parasuis and vtaA. The cut-off threshold cycle was found to be at 35. The developed sensitive and specific qPCR assay has the potential to become a useful molecular tool, which could be implemented in veterinary diagnostic laboratories for the detection and identification of G. parasuis, its virulence marker vtaA, M. hyorhinis, and M. hyosynoviae.


Assuntos
Reação em Cadeia da Polimerase Multiplex , Infecções por Mycoplasma , Mycoplasma hyorhinis , Mycoplasma hyosynoviae , Infecções por Pasteurellaceae , Pasteurellaceae , Doenças dos Suínos , Reação em Cadeia da Polimerase Multiplex/métodos , Pasteurellaceae/isolamento & purificação , Mycoplasma hyorhinis/isolamento & purificação , Mycoplasma hyosynoviae/isolamento & purificação , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/microbiologia , Animais , Suínos , Infecções por Mycoplasma/diagnóstico , Infecções por Mycoplasma/microbiologia , Infecções por Mycoplasma/veterinária , Infecções por Pasteurellaceae/diagnóstico , Infecções por Pasteurellaceae/microbiologia , Infecções por Pasteurellaceae/veterinária , Projetos Piloto , Sensibilidade e Especificidade
9.
Vet Res Commun ; 47(2): 683-691, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-36342628

RESUMO

The purpose of this report is to provide information about the different presentations of cardiac and extra-cardiac histophilosis and, to assess the antimicrobial (ATM) susceptibility of Histophilus somni isolated from these cardiac lesions to different ATM agents commonly used for treating bovine bacterial respiratory pathogens. Eight feedlot calves, which died after suffering from food rejection, apathy, hyperthermia, cough and nasal mucous discharge, and lack of response to ATM therapy, were studied. Cardiac lesions observed at necropsy included valvular/mural endocarditis, myocardial infarction, and necrotizing myocarditis, miliar non-suppurative myocarditis, myocardic necrotic sequestrum, and/or pericarditis. Histopathological, bacteriological and molecular studies confirmed the presence of a fastidious microorganism in the affected organs. H. somni showed no resistance to most ATM tested (ceftiofur, gamithromycin, enrofloxacin, florfenicol, tilmicosin). The results obtained in this study confirmed that H. somni was the main cause of the subacute cardiac lesions associated with hyperthermia, apathy and respiratory signs observed in cattle examined in this research. These presentations must be considered by veterinary practitioners in order to establish a rational therapeutic.


Assuntos
Doenças dos Bovinos , Miocardite , Infecções por Pasteurellaceae , Pasteurellaceae , Bovinos , Animais , Doenças dos Bovinos/microbiologia , Infecções por Pasteurellaceae/veterinária , Infecções por Pasteurellaceae/microbiologia , Miocardite/microbiologia , Miocardite/veterinária , Morte
10.
Clin Neurol Neurosurg ; 219: 107337, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35717764

RESUMO

BACKGROUND: Aggregatibacter aphrophilus(A. aphrophilus)is one of the organisms of the HACEK group. Previously reported cases of brain abscesses caused by A. aphrophilus infection have occurred in children with a basis for congenital heart disease, or in adults with a basis for dental disease. Rare cases of brain abscess caused by A. aphrophilus have been reported in adults with congenital heart disease or in patients without dental disease history. Herein we present a rare case of brain abscess caused by A. aphrophilus, who was in association with atrial septal defect for more than 20 years, and had no dental disease and did not develop infective endocarditis. CASE PRESENTATION: A 51-year-old female was admitted due to progressively worsening headache and left limb weakness for more than 10 days. She denied the history of chronic diseases such as hypertension and diabetes, and no periodontal disease. While she had a history of atrial septal defect, a form of congenital heart disease with severe pulmonary hypertension for more than 20 years. After admission, echocardiographic illustrated congenital heart disease with severe pulmonary hypertension. CT and MRI showed brain abscess. Cerebrospinal fluid (CSF) results also confirmed the presence of intracranial infection. Empirical therapy with vancomycin 1.0 g i.v q12h and meropenem 2.0 g i.v q8h was initiated from the day of admission. On the fourth day after admission, brain abscess resection and decompressive craniectomy were performed, and the pus drained on operation were cultured and Gram-negative bacilli grew, which was identified as A.aphrophilus. Vancomycin was discontinued and meropenem was continued(2.0 g i.v q8h)for 5 weeks, followed by oral levofloxacin 0.5 qd for 4 weeks of out-patient antibiotics. The patient recovered fully within 9 weeks of treatment. CONCLUSIONS: This is the first case of A. aphrophilus to cause brain abscess in adult with a history of congenital heart disease for more than 20 years, who had no dental disease and did not develop infective endocarditis. We also highlight the value of bacterial 16 S rDNA PCR amplification and sequencing in identifying bacteria in abscesses which are culture-negative, and prompt surgical treatment,choosing effective antibiotics and appropriate course of treatment will get better clinical effect.


Assuntos
Aggregatibacter aphrophilus , Abscesso Encefálico , Endocardite , Cardiopatias Congênitas , Comunicação Interatrial , Hipertensão Pulmonar , Infecções por Pasteurellaceae , Adulto , Antibacterianos/uso terapêutico , Abscesso Encefálico/diagnóstico por imagem , Abscesso Encefálico/tratamento farmacológico , Abscesso Encefálico/cirurgia , Criança , Endocardite/complicações , Endocardite/tratamento farmacológico , Feminino , Cardiopatias Congênitas/complicações , Cardiopatias Congênitas/tratamento farmacológico , Comunicação Interatrial/complicações , Comunicação Interatrial/tratamento farmacológico , Comunicação Interatrial/cirurgia , Humanos , Hipertensão Pulmonar/complicações , Hipertensão Pulmonar/tratamento farmacológico , Meropeném/uso terapêutico , Pessoa de Meia-Idade , Infecções por Pasteurellaceae/complicações , Infecções por Pasteurellaceae/tratamento farmacológico , Infecções por Pasteurellaceae/microbiologia , Vancomicina/uso terapêutico
11.
Biochem Biophys Res Commun ; 595: 76-81, 2022 03 05.
Artigo em Inglês | MEDLINE | ID: mdl-35101666

RESUMO

L-enantiomers of antimicrobial peptides (AMPs) are sensitive to proteolytic degradation; however, D-enantiomers of AMPs are expected to provide improved proteolytic resistance. The present study aimed to comparatively investigate the in vitro antibacterial activity, trypsin and serum stability, toxicity, and in vivo antibacterial activity of L-enantiomeric bovine NK2A (L-NK2A) and its D-enantiomeric NK2A (D-NK2A). Circular dichroism spectroscopy of D-NK2A and L-NK2A in anionic liposomes showed α-helical structures and the α-helical conformation of D-NK2A was a mirror image of L-NK2A. Both D-NK2A and L-NK2A displayed minimal in vitro and in vivo toxicities. RP-HPLC and mass spectrometry analyses revealed that D-NK2A, but not L-NK2A, was resistant to trypsin digestion. D-NK2A and L-NK2A showed similar in vitro bacterial killing activities against Histophilus somni. Slightly reduced antibacterial activity was observed when D-NK2A and L-NK2A were pre-incubated with serum. Confocal and transmission electron microscopic findings confirmed that both peptides induced disruption of bacterial inner- and outer-membranes. Improved survivals with D-NK2A treatment were observed when compared to L-NK2A in a murine model of acute H. somni septicemia. We conclude that antibacterial activity and mode of action of NK2A are not chiral specific. With further optimization, D-NK2A may be a viable AMP candidate to combat bacterial infections.


Assuntos
Antibacterianos/farmacologia , Peptídeos Antimicrobianos/farmacologia , Infecções por Pasteurellaceae/prevenção & controle , Pasteurellaceae/efeitos dos fármacos , Proteolipídeos/farmacologia , Animais , Antibacterianos/química , Peptídeos Antimicrobianos/química , Bovinos , Dicroísmo Circular , Estimativa de Kaplan-Meier , Camundongos , Microscopia Eletrônica de Transmissão , Pasteurellaceae/fisiologia , Pasteurellaceae/ultraestrutura , Infecções por Pasteurellaceae/microbiologia , Estabilidade Proteica , Estrutura Secundária de Proteína , Proteolipídeos/química , Estereoisomerismo
12.
Toxins (Basel) ; 14(1)2022 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-35051027

RESUMO

Leukotoxin A (LtxA) is the major virulence factor of an oral bacterium known as Aggregatibacter actinomycetemcomitans (Aa). LtxA is associated with elevated levels of anti-citrullinated protein antibodies (ACPA) in rheumatoid arthritis (RA) patients. LtxA targets leukocytes and triggers an influx of extracellular calcium into cytosol. The current proposed model of LtxA-mediated hypercitrullination involves the dysregulated activation of peptidylarginine deiminase (PAD) enzymes to citrullinate proteins, the release of hypercitrullinated proteins through cell death, and the production of autoantigens recognized by ACPA. Although model-based evidence is yet to be established, its interaction with the host's immune system sparked interest in the role of LtxA in RA. The first part of this review summarizes the current knowledge of Aa and LtxA. The next part highlights the findings of previous studies on the association of Aa or LtxA with RA aetiology. Finally, we discuss the unresolved aspects of the proposed link between LtxA of Aa and RA.


Assuntos
Aggregatibacter actinomycetemcomitans/fisiologia , Artrite Reumatoide/microbiologia , Infecções por Pasteurellaceae/microbiologia , Anticorpos Antiproteína Citrulinada/imunologia , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Exotoxinas/imunologia , Humanos , Infecções por Pasteurellaceae/imunologia , Infecções por Pasteurellaceae/patologia
13.
Microbiol Spectr ; 9(3): e0143121, 2021 12 22.
Artigo em Inglês | MEDLINE | ID: mdl-34851158

RESUMO

Histophilus somni is a Gram-negative bacterial organism that acts as an opportunistic pathogen and is a fastidious member of the Pasteurellaceae family associated with diseases of respiratory, reproductive, cardiac, and other tissues of ruminants. We identified an intervening sequence (IVS) embedded in all five copies of the 23S rRNA gene in the closed genome sequence of the H. somni isolate USDA-ARS-USMARC-63250 that may play an important role in affecting the biology of the organism. Sequencing the RNA from this isolate shows that most of the IVS is cleaved from the transcript, resulting in independent fragments of this structural rRNA that remain functional within the bacterial ribosome. The IVS lies between positions 1170 and 1278 bp of the 3,017-bp gene and exhibits self-complementarity between its 5' and 3' ends that predicts a stem-loop structure interrupting helix-45 in the transcribed 23S rRNA. Excision removes a 94-nucleotide (nt) stem-loop structure that displays an unusual 1-nt 3' end overhang instead of the more typical 2-nt overhang commonly observed at the ends of other excised IVS stem-loops. A comparison with genomes of other H. somni isolates indicates that this IVS is highly conserved, with 31 of 32 complete genomes having similar interruptions of canonical 23S rRNA genes. The potential biological effects of either the released IVS or the fragmentation of the functional 23S rRNA are unknown, but fragmentation may enhance rRNA degradation in ways that contribute to the regulation of gene expression. IMPORTANCE The genome biology underlying H. somni virulence, pathogenicity, environmental adaptability, and broad tissue tropism is understood poorly. We identified a novel H. somni 109-nt IVS stem-loop structure, of which the central portion is excised from the 23S rRNA transcript, resulting in the fragmentation of this rRNA in the H. somni isolate USDA-ARS-USMARC-63250 and the release of a 94-nt structured RNA of unknown function. We determined that this peculiar rRNA biology is widespread among sequenced H. somni isolates, suggesting it has importance to organism biology. The fragmented 23S rRNA molecules remain functional in the ribosome, given that the isolate grows in culture. The structured excised portion of the IVS, presumably due to the action of the endoribonuclease III, has an unusual 1-nt 3' end overhang. This newly discovered H. somni 23S rRNA fragmentation may enhance rRNA degradation providing a previously unrecognized avenue for regulating H. somni biological processes.


Assuntos
Sequências Repetidas Invertidas/genética , Conformação de Ácido Nucleico , Infecções por Pasteurellaceae/veterinária , Pasteurellaceae/genética , RNA Ribossômico 23S/genética , Animais , Sequência de Bases/genética , Bovinos , Doenças dos Bovinos/microbiologia , Íntrons/genética , RNA Bacteriano/genética , Infecções Respiratórias/microbiologia , Infecções Respiratórias/veterinária , Ribossomos/genética , Análise de Sequência de RNA
14.
J Microbiol Methods ; 191: 106360, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34740721

RESUMO

Glaesserella australis, a newly described bacterial species, has been isolated from pig lungs that displayed lesions very similar to those caused by Actinobacillus pleuropneumoniae, prompting the need for a validated diagnostic tool. In this work, we have altered a multiplex PCR used for the identification of cultures of G. australis, A. pleuropneumoniae and Pasteurella multocida to be more sensitive and then evaluated the use of the altered diagnostic tool on cultures and directly on tissues. The altered multiplex PCR was validated using 47 related species, both type/reference strains and field isolates. The sensitivity was assessed by serial dilutions and used a mixture of target bacteria in different concentrations. Further, 166 lung samples from 54 farms from four Australian States were used to validate the ability of the multiplex PCR to detect bacteria in lung swabs. The multiplex PCR was specific for the three target species. The assay could detect a minimum of 40 colony forming units (CFU) of G. australis, 786 CFU of A. pleuropneumoniae and 238 CFU of P. multocida. The multiplex PCR yielded more positives than coventional bacteriological examination. From a total of 166 lung samples, 51.9%, 51.9% and 5.6% of farms were PCR positive for P. multocida, A. pleuropneumoniae and G. australis, respectively. The results suggested that the new multiplex PCR was specific, sensitive and out performed traditional culture. The prevalence of G. australis was not very high, but it was the dominant pathogen in infected pigs.


Assuntos
Actinobacillus pleuropneumoniae/genética , Reação em Cadeia da Polimerase Multiplex/métodos , Pasteurella multocida/genética , Pasteurellaceae/genética , Infecções por Actinobacillus/diagnóstico , Infecções por Actinobacillus/epidemiologia , Infecções por Actinobacillus/veterinária , Actinobacillus pleuropneumoniae/isolamento & purificação , Animais , Testes Diagnósticos de Rotina , Fazendas , Pulmão/microbiologia , Infecções por Pasteurella/diagnóstico , Infecções por Pasteurella/epidemiologia , Infecções por Pasteurella/veterinária , Pasteurella multocida/isolamento & purificação , Pasteurellaceae/isolamento & purificação , Infecções por Pasteurellaceae/diagnóstico , Infecções por Pasteurellaceae/epidemiologia , Infecções por Pasteurellaceae/veterinária , Prevalência , Sensibilidade e Especificidade , Células-Tronco , Suínos , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/epidemiologia , Doenças dos Suínos/microbiologia
15.
Vet Res ; 52(1): 135, 2021 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-34674760

RESUMO

Glaesserella parasuis (G. parasuis) is a commensal bacterium in the upper respiratory tract of pigs that can also cause the swine Glässer disease, which induces an intensive inflammatory response and results in significant economic losses to the swine industry worldwide. G. parasuis can cause disease through infection of the respiratory tract, resulting in systemic infection, but the mechanism is largely unknown. Recently we showed that Glaesserella parasuis serotype 4 (GPS4) increased swine tracheal epithelial barrier permeability, resulting in easier bacterial translocation. Tight junction proteins (TJ) play a crucial role in maintaining the integrity and impermeability of the epithelial barrier. GPS4 decreased the expression of the TJ ZO-1 and occludin in swine tracheal epithelial cells (STEC). Furthermore, the proinflammatory cytokines IL-6, IL-8 and TNF-α were significantly upregulated in GPS4-infected STEC, and both the MAPK and NF-κB signaling pathways were activated and contributed to the expression of TNF-α. We demonstrate that the production of proinflammatory cytokines, especially TNF-α, during GPS4 infection was involved in barrier dysfunction. Additionally, animal challenge experiments confirmed that GPS4 infection downregulated TJ in the lungs of piglets and induced a severe inflammatory response. In general, G. parasuis infection downregulated the expression of TJ and induced massive secretion of proinflammatory cytokines, resulting in epithelial barrier disruption and favoring bacterial infection. This study allowed us to better understand the mechanism by which G. parasuis crosses the respiratory tract of pigs.


Assuntos
Translocação Bacteriana , Haemophilus parasuis/fisiologia , Infecções por Pasteurellaceae/veterinária , Transdução de Sinais , Doenças dos Suínos/microbiologia , Animais , Células Epiteliais , Infecções por Haemophilus/microbiologia , Infecções por Haemophilus/fisiopatologia , Infecções por Haemophilus/veterinária , Haemophilus parasuis/genética , Infecções por Pasteurellaceae/microbiologia , Infecções por Pasteurellaceae/fisiopatologia , Sorogrupo , Sus scrofa , Suínos , Doenças dos Suínos/fisiopatologia
16.
Vet Res ; 52(1): 126, 2021 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-34600578

RESUMO

This work modifies a loop-mediated isothermal amplification (LAMP) assay to detect the bovine respiratory disease (BRD) bacterial pathogens Pasteurella multocida, Mannheimia haemolytica, and Histophilus somni in a colorimetric format on a farm. BRD causes a significant health and economic burden worldwide that partially stems from the challenges involved in determining the pathogens causing the disease. Methods such as polymerase chain reaction (PCR) have the potential to identify the causative pathogens but require lab equipment and extensive sample processing making the process lengthy and expensive. To combat this limitation, LAMP allows accurate pathogen detection in unprocessed samples by the naked eye allowing for potentially faster and more precise diagnostics on the farm. The assay developed here offers 66.7-100% analytical sensitivity, and 100% analytical specificity (using contrived samples) while providing 60-100% concordance with PCR results when tested on five steers in a feedlot. The use of a consumer-grade water bath enabled on-farm execution by collecting a nasal swab from cattle and provided a colorimetric result within 60 min. Such an assay holds the potential to provide rapid pen-side diagnostics to cattle producers and veterinarians.


Assuntos
Doenças dos Bovinos/diagnóstico , Colorimetria/veterinária , Testes Diagnósticos de Rotina/veterinária , Técnicas de Diagnóstico Molecular/veterinária , Técnicas de Amplificação de Ácido Nucleico/veterinária , Infecções por Pasteurellaceae/veterinária , Pasteurellaceae/isolamento & purificação , Animais , Bovinos , Doenças dos Bovinos/microbiologia , Colorimetria/instrumentação , Testes Diagnósticos de Rotina/instrumentação , Mannheimia haemolytica/isolamento & purificação , Técnicas de Diagnóstico Molecular/instrumentação , Nariz/microbiologia , Técnicas de Amplificação de Ácido Nucleico/instrumentação , Infecções por Pasteurella/diagnóstico , Infecções por Pasteurella/microbiologia , Infecções por Pasteurella/veterinária , Pasteurella multocida/isolamento & purificação , Infecções por Pasteurellaceae/diagnóstico , Infecções por Pasteurellaceae/microbiologia
17.
Front Immunol ; 12: 707096, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34456916

RESUMO

Aggregatibacter actinomycetemcomitans is a gram-negative facultative anaerobe and an opportunistic oral pathogen, strongly associated with periodontitis and other inflammatory diseases. Periodontitis is a chronic inflammation of the periodontium resulting from the inflammatory response of the host towards the dysbiotic microbial community present at the gingival crevice. Previously, our group identified catecholamines and iron as the signals that activate the QseBC two-component system in A. actinomycetemcomitans, necessary for the organism to acquire iron as a nutrient to survive in the anaerobic environment. However, the source of catecholamines has not been identified. It has been reported that mouse neutrophils can release catecholamines. In periodontitis, large infiltration of neutrophils is found at the subgingival pocket; hence, we wanted to test the hypothesis that A. actinomycetemcomitans exploits human neutrophils as a source for catecholamines. In the present study, we showed that human neutrophils synthesize, store, and release epinephrine, one of the three main types of catecholamines. Human neutrophil challenge with A. actinomycetemcomitans induced exocytosis of neutrophil granule subtypes: secretory vesicles, specific granules, gelatinase granules, and azurophilic granules. In addition, by selectively inhibiting granule exocytosis, we present the first evidence that epinephrine is stored in azurophilic granules. Using QseC mutants, we showed that the periplasmic domain of the QseC sensor kinase is required for the interaction between A. actinomycetemcomitans and epinephrine. Finally, epinephrine-containing supernatants collected from human neutrophils promoted A. actinomycetemcomitans growth and induced the expression of the qseBC operon under anaerobic conditions. Based on our findings, we propose that A. actinomycetemcomitans promotes azurophilic granule exocytosis by neutrophils as an epinephrine source to promote bacterial survival.


Assuntos
Aggregatibacter actinomycetemcomitans/metabolismo , Epinefrina/metabolismo , Neutrófilos/metabolismo , Infecções por Pasteurellaceae/metabolismo , Periodontite/microbiologia , Sobrevivência Celular/fisiologia , Humanos
18.
Avian Dis ; 65(1): 95-101, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-34339129

RESUMO

This is the first extensive report on the identification and characterization of Avibacterium paragallinarum (AVP) isolates obtained from outbreaks of infectious coryza (IC) in IC-vaccinated layer flocks from Sonora State in Mexico. Isolates obtained from IC outbreaks during the years 2007, 2014, 2015, 2017, and 2019 were identified by conventional PCR test and 16S rRNA gene analysis, serotyped by Page serotyping and genotyped by the recently described partial sequence analysis of the HPG2 region. Furthermore, antimicrobial susceptibility profiles were determined by a recently improved minimal inhibitory concentration (MIC) test. The conventional PCR test and the 16S rRNA analyses confirmed the isolates as AVP. Serotyping results showed the involvement of isolates belonging to serotypes A, B, and C in the IC outbreaks. Genotyping of the HPG2 region revealed the presence of sequence type (ST)1, ST4, and ST11, of which the latter has also been identified in Europe. The MIC susceptibility test showed that all tested isolates were susceptible for the majority of tested antimicrobials, including erythromycin and tetracycline, which are important antibiotics for the treatment of IC. The IC situation in Sonora State, Mexico, is complex because of the presence of serotypes A, B, and C. This finding emphasizes the importance of biosecurity in combination with the application of the most optimal vaccination programs in the control of IC in Sonora State, Mexico.


Nota de investigación­Análisis de secuencias de la región HPG2 y susceptibilidad antimicrobiana de aislamientos de Avibacterium paragallinarum obtenidos de brotes de coriza infecciosa en aves de postura comerciales en el estado de Sonora, México. Este es el primer informe extenso sobre la identificación y caracterización de aislamientos de Avibacterium paragallinarum (AVP) obtenidos de brotes de coriza infecciosa (IC) de parvadas de ponedoras vacunadas con coriza infecciosa en el estado de Sonora en México. Los aislamientos obtenidos de los brotes de coriza infecciosa durante los años 2007, 2014, 2015, 2017 y 2019 se identificaron mediante una prueba de PCR convencional y el análisis del gene de ARNr 16S, se serotipificaron mediante el método de Page y se genotipificaron mediante el análisis parcial de secuencias descrito recientemente de la región HPG2. Además, se determinaron los perfiles de susceptibilidad a los antimicrobianos mediante la prueba de concentración mínima inhibitoria (MIC) que ha sido mejorada recientemente. La prueba de PCR convencional y los análisis de secuencias del gene ARNr 16S confirmaron que los aislados eran A. paragallinarum. Los resultados de la serotipificación mostraron la participación de aislamientos pertenecientes a los serotipos A, B y C en los brotes de coriza infecciosa. La genotipificación de la región HPG2 reveló la presencia de secuencias del tipo (ST) 1, ST4 y ST11, de los cuales este último también ha sido identificada en Europa. La prueba de susceptibilidad por concentración mínima inhibitoria mostró que todos los aislados analizados eran susceptibles a la mayoría de los antimicrobianos analizados, incluida la eritromicina y la tetraciclina, que son antibióticos importantes para el tratamiento contra la coriza infecciosa. La situación de coriza infecciosa en el estado de Sonora, México, es compleja por la presencia de los serotipos A, B y C. Este hallazgo enfatiza la importancia de la bioseguridad en combinación con la aplicación de los programas de vacunación óptimos en el control de la coriza infecciosa en el estado de Sonora, México.


Assuntos
Galinhas , Farmacorresistência Bacteriana , Infecções por Pasteurellaceae/veterinária , Pasteurellaceae/isolamento & purificação , Doenças das Aves Domésticas , Proteínas Virais/análise , Animais , Feminino , México , Testes de Sensibilidade Microbiana/veterinária , Pasteurellaceae/efeitos dos fármacos , Pasteurellaceae/genética , Infecções por Pasteurellaceae/diagnóstico , Infecções por Pasteurellaceae/microbiologia , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/microbiologia
20.
DNA Cell Biol ; 40(6): 776-790, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34029124

RESUMO

Glaesserella parasuis causes porcine Glässer's disease and lipopolysaccharide (LPS) induces acute inflammation and pathological damage. Baicalin has antioxidant, antimicrobial, and anti-inflammatory functions. Long noncoding RNAs (lncRNAs) play key regulatory functions during bacterial infection. However, the role of lncRNAs in the vascular dysfunction induced by a combination of G. parasuis and LPS during systemic inflammation and the effect of baicalin on lncRNA expression induced in porcine aortic vascular endothelial cells (PAVECs) by a combination of G. parasuis and LPS have not been investigated. In this study, we investigated the changes in lncRNA and mRNA expression induced in PAVECs by G. parasuis, LPS, or a combination of G. parasuis and LPS, and the action of baicalin on lncRNA expression induced in PAVECs by the combination of G. parasuis and LPS. Our results showed 133 lncRNAs and 602 genes were differentially expressed when PAVECs were stimulated with the combination of G. parasuis and LPS, whereas 107 lncRNAs and 936 genes were differentially expressed when PAVECs were stimulated with the combination of G. parasuis and LPS after pretreatment with baicalin. The Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed the dominant signaling pathways triggered by the combination of G. parasuis and LPS were the tumor necrosis factor signaling pathway, phosphatidylinositol signaling system, and inositol phosphate metabolism. Protein-protein interaction network analysis showed the differentially expressed target genes of the differentially expressed lncRNAs (DELs) were related to each other. A coexpression analysis indicated the expression levels of the DELs were co-regulated with those of their differentially expressed target genes. This is the first study to systematically compare the changes in lncRNAs and mRNAs in PAVECs stimulated with a combination of G. parasuis and LPS. Our data clarified the mechanisms underlying the vascular inflammation and damage triggered by G. parasuis and LPS, and it may provide novel targets for the treatment of LPS-induced systemic inflammation.


Assuntos
Anti-Inflamatórios , Células Endoteliais , Flavonoides , Inflamação , Infecções por Pasteurellaceae/veterinária , Doenças dos Suínos/tratamento farmacológico , Animais , Anti-Inflamatórios/farmacologia , Anti-Inflamatórios/uso terapêutico , Linhagem Celular , Células Endoteliais/efeitos dos fármacos , Células Endoteliais/patologia , Flavonoides/farmacologia , Flavonoides/uso terapêutico , Inflamação/tratamento farmacológico , Inflamação/veterinária , Pasteurellaceae , Infecções por Pasteurellaceae/tratamento farmacológico , RNA Longo não Codificante , RNA Mensageiro/genética , Suínos , Doenças dos Suínos/microbiologia , Transcriptoma
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