ABSTRACT
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.
Subject(s)
Abomasum , Mannheimia haemolytica , Necrosis , Pasteurellaceae Infections , Sheep Diseases , Animals , Mannheimia haemolytica/isolation & purification , Sheep Diseases/pathology , Sheep Diseases/microbiology , Sheep , Abomasum/pathology , Abomasum/microbiology , Pasteurellaceae Infections/veterinary , Pasteurellaceae Infections/pathology , Pasteurellaceae Infections/microbiology , Necrosis/veterinary , Necrosis/pathology , Necrosis/microbiology , Stomach Diseases/veterinary , Stomach Diseases/pathology , Stomach Diseases/microbiology , Male , Female , Immunohistochemistry/veterinaryABSTRACT
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.
Subject(s)
Aggregatibacter actinomycetemcomitans/physiology , Arthritis, Rheumatoid/microbiology , Pasteurellaceae Infections/microbiology , Anti-Citrullinated Protein Antibodies/immunology , Arthritis, Rheumatoid/immunology , Arthritis, Rheumatoid/pathology , Exotoxins/immunology , Humans , Pasteurellaceae Infections/immunology , Pasteurellaceae Infections/pathologyABSTRACT
Case history: In July and August 2019, 15/40, ≤48-hour-old calves became acutely ill. The calves were all born on-farm, transferred to pens soon after birth, and fed with "gold" colostrum. The hygiene, biosecurity and ventilation in the pens were poor. Of the 15 calves, 11 died or were euthanised and four calves, ≤48-hour-old, that became acutely ill later in the outbreak were treated with cefquinome, a fourth-generation cephalosporin, and recovered. Clinical findings: The affected calves presented with acute recumbency, lethargy, tachypnoea, tachycardia, increased lung sounds, inability to stand or feed, and dehydration without pyrexia. Pathological findings: Gross findings in a calf that died naturally included fibrinous pleuropneumonia, marked oedematous expansion of the interlobular septa, especially in the ventral lung lobes, fibrinous polyserositis and fibrinous polyarthritis. A second calf that was euthanised had strikingly similar lung lesions. Histologically, the pulmonary interlobular septa of both calves were prominently expanded by oedema, dilated lymphatics and the infiltration of numerous neutrophils and macrophages interspersed with small Gram-negative rod bacteria. Likewise, the visceral pleura showed fibrinopurulent inflammation with numerous small Gram-negative rods. Microbiological findings: Microbial culture and matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry identified Bibersteinia trehalosi in the lung, stifle joint and peritoneal cavity of the first calf and lung of the second. Diagnosis: B. trehalosi acute fibrinous pleuropneumonia and septicaemia. Clinical relevance: This is the first report of the clinical findings and histological lesions of B. trehalosi pleuropneumonia and septicaemia in calves in New Zealand. The pathogen is isolated with increasing frequency from cases of bovine respiratory disease in dairy cows, feedlot cattle and calves in the United Kingdom and North America. The importance of microbial culture in cases such as this with unusual lung lesions in calves <48 hours of age, cannot be over emphasised. Cefquinome was administered to all remaining heifer calves and four calves that became ill later in the outbreak recovered after cefquinome treatment.
Subject(s)
Cattle Diseases/microbiology , Pasteurellaceae Infections/veterinary , Pasteurellaceae/isolation & purification , Pleuropneumonia/veterinary , Sepsis/veterinary , Animals , Animals, Newborn , Cattle , Lung/pathology , New Zealand , Pasteurellaceae Infections/pathology , Pleuropneumonia/microbiology , Pleuropneumonia/pathology , Sepsis/microbiologyABSTRACT
Aggregatibacter actinomycetemcomitans is frequently isolated from localized aggressive periodontitis and periodontitis associated with systemic diseases. A. actinomycetemcomitans produces a leukotoxin, which induces apoptosis in human leukocytes. The leukotoxin expression is dependent on the upstream sequence, likely including the promoter, of the gene encoding leukotoxin; strains with the truncated/short upstream sequence express more leukotoxin than strains with the general/long upstream. This chapter addresses the determination of the type of the leukotoxin promoter by PCR analysis, and detection of the apoptosis in the coculture of human monocyte cell line (THP-1) with A. actinomycetemcomitans by the DNA ladder formation, membrane perturbation, and lactate dehydrogenase release.
Subject(s)
Aggregatibacter actinomycetemcomitans/pathogenicity , Exotoxins/metabolism , Leukocytes/pathology , Pasteurellaceae Infections/pathology , Aggregatibacter actinomycetemcomitans/genetics , Aggregatibacter actinomycetemcomitans/metabolism , Apoptosis , Cell Line , Coculture Techniques/methods , Exotoxins/genetics , Humans , Leukocytes/metabolism , Leukocytes/microbiology , Pasteurellaceae Infections/microbiology , Polymerase Chain Reaction/methods , Promoter Regions, Genetic , VirulenceABSTRACT
This study investigated the association of Pasteurella multocida isolation and the molecular characteristics of the isolates with the presence of pneumonic lesions in lambs at slaughter to assess its importance as a causative agent of pneumonic pasteurellosis compared with Mannheimia haemolytica. P. multocida was isolated from the 13.9% and 2.7%, and M. haemolytica from the 36.4% and 26.8%, of lungs with and without lesions, respectively (P < 0.05). Both microorganisms were frequently coisolated (23.2% and 12.5% from lungs with and without lesions, respectively). Isolation of P. multocida alone exhibited greater strength of association with pneumonic lesions (OR 11.4; 95% CI 3.2-40.6) than that exhibited by M. haemolytica alone (OR 3.0; 95% CI 1.6-5.4). Cluster analysis grouped the lungs into four clusters characterized by the isolation of M. haemolytica or P. multocida alone (clusters 1 and 4), coisolation of both microorganisms (cluster 3), and isolation of neither (cluster 2). Cluster 4 lungs exhibited higher frequencies of pneumonic lesions (87.5%) and severe (20.8%) and moderate (25.0%) lesions. Lungs coinfected with both pathogens (cluster 3) did not exhibit a higher frequency of severe and moderate consolidation lesions (6.1% and 14.3%, respectively), suggesting that P. multocida and M. haemolytica do not act synergically to cause more severe pneumonic infections. The greater strength of association of P. multocida isolation with pneumonic lesions together with the higher severity of the lesions caused could indicate a greater role played by this pathogen in the aetiopathogenesis of pneumonic pasteurellosis in sheep than is commonly assumed.
Subject(s)
Coinfection/veterinary , Lung/pathology , Pasteurellosis, Pneumonic/microbiology , Pasteurellosis, Pneumonic/pathology , Sheep Diseases/microbiology , Abattoirs , Animals , Coinfection/microbiology , Farms , Lung/microbiology , Mannheimia haemolytica/pathogenicity , Pasteurella multocida/isolation & purification , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Severity of Illness Index , Sheep , Sheep Diseases/pathology , Spain , Species SpecificityABSTRACT
Carcinogenesis is caused by genome instability, one of the major causes of which is double-strand DNA breaks (DSBs). Interestingly, infection by particular species of bacteria can induce DSBs in host cells. For example, several reports suggest an association between periodontal disease and oral cancer. Aggregatibacter actinomycetemcomitans, a common periodontal pathogen, causes DSBs in the host cell. Pulsed-field gel electrophoresis (PFGE) is often used to identify DSBs in host cells. However, as established during investigation of A. actinomycetemcomitans infection, it is often difficult to determine whether broken DNA fragments are indeed from human chromosomes or whether they are bacterial in origin using PFGE-based methods. Because the method involves the coculture of human cells with bacteria, both bacterial and human DNA fragments may be present in the broken DNA fraction. To address this problem, we have developed a method to detect only human chromosomal DNA upon PFGE analysis. Human chromosomes were prelabeled with halogenated deoxyuridine (e.g., BrdU and IdU) before being fractionated by PFGE and visualized by immunoblotting. As proof of concept, we successfully used this method to investigate the mechanism of DSB formation in host chromosomes following infection with genotoxic bacterial species.
Subject(s)
Aggregatibacter actinomycetemcomitans , Chromosomes, Human/metabolism , DNA Breaks, Double-Stranded , Electrophoresis, Gel, Pulsed-Field , Pasteurellaceae Infections/metabolism , Cell Line , Chromosomes, Human/chemistry , Humans , Pasteurellaceae Infections/pathologyABSTRACT
The prevalence of Gallibacterium anatis in poultry production has increased over the last two decades. However, only a few studies have explored the pathogenicity of this bacterium in commercial layer chickens. This trial studied the aspects of the pathogenicity of a Gallibacterium anatis biovar haemolytica local Egyptian isolate (previously registered as strain B14 with GenBank accession no. KJ026147). We used 500 base pairs of a 16S ribosomal RNA gene and the 16S-23S ribosomal RNA intergenic spacer, partial sequence in an experimental infection trial in commercial White Shaver layer chickens aged 19 wk. The hens were divided into three groups of 40 birds each. The hens in Groups 1 and 2 were experimentally infected through the intranasal (IN) and intravenous (IV) routes, respectively, with a dose of 0.2 ml/bird containing 1.2 × 109 colony-forming units/ml. In contrast, Group 3 was kept as a noninfected control group. Both IN and IV infections resulted in a delayed egg laying for 1 wk and a significant (P ≤ 0.05) drop in egg production by 7.81% and 10.28% compared with the control group over 7 wk. Severe lesions in the form of hemorrhagic pneumonia, catarrhal tracheitis, ovarian follicle and oviductal regression, and septicemia were evident on necropsy, demonstrating the pathogenicity of G. anatis as a primary pathogen.
Subject(s)
Chickens , Ovarian Diseases/veterinary , Pasteurellaceae Infections/veterinary , Pasteurellaceae/physiology , Pasteurellaceae/pathogenicity , Poultry Diseases/pathology , Respiratory Tract Diseases/veterinary , Animals , Female , Ovarian Diseases/microbiology , Ovarian Diseases/pathology , Ovarian Diseases/physiopathology , Pasteurellaceae/genetics , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Pasteurellaceae Infections/physiopathology , Poultry Diseases/microbiology , Poultry Diseases/physiopathology , Respiratory Tract Diseases/microbiology , Respiratory Tract Diseases/pathology , Respiratory Tract Diseases/physiopathology , Sepsis/microbiology , Sepsis/pathology , Sepsis/physiopathology , Sepsis/veterinaryABSTRACT
Objective was the differential quantitative proteomics study of ovine mastitis induced by Mannheimia haemolytica; clinical, microbiological, cytological and histopathological methods were employed for confirmation and monitoring. Proteins were separated by two-dimensional gel electrophoresis (2-DE) for all samples and differentially abundant proteins were identified by mass spectrometry; comparisons were performed with pre- (blood, milk) and post- (milk of contralateral gland) inoculation findings. Animals developed mastitis, confirmed by isolation of challenge strain and increase of neutrophils in milk and by histopathological evidence. In blood plasma, 33 differentially abundant proteins (compared to findings before challenge) were identified: 6 with decrease, 13 with new appearance and 14 with varying abundance. In a post-challenge milk whey protein reference map, 65 proteins were identified; actin cytoplasmic-1, beta-lactoglobulin-1/B, cathelicidin-1 predominated. Further, 89 differentially abundant proteins (compared to findings before challenge) were identified: 18 with decrease, 53 with new appearance, 3 with increase and 15 with varying abundance; 15 proteins showed status changes in blood plasma and milk whey. Differential abundance from inoculated and contralateral glands revealed 74 proteins only from the inoculated gland. Most differentially abundant proteins in milk whey were involved in cell organisation and biogenesis (nâ¯=â¯17) or in inflammatory and defence response (nâ¯=â¯13). SIGNIFICANCE: The proteomes of blood and milk from ewes with experimental mastitis caused by Mannheimia haemolytica and the differential proteomics in sequential samples after challenge are presented for the first time. This is the first detailed proteomics study in M. haemolytica-associated mastitis in ewes. An experimental model fully simulating natural mastitis has been used. Use of experimentally induced mastitis minimised potential variations and allowed consistency of results. The study included evaluation of changes in blood plasma and milk whey. Protein patterns have been studied, indicating with great accuracy changes that had occurred as part of the disease process and development, during the acute phase of infection. Relevant protein-protein interactions were studied. The entirety of proteomics findings has suggested that affected ewes had mounted a defence response that had been regulated by many proteins (e.g., cathelicidins, haptoglobin, serum amyloid A) and through various pathways (e.g., acute phase response, binding and transporting significant ions and molecules); these were interdependent at various points. Potential biomarkers have been indicated for use in diagnostic assays of mastitis.
Subject(s)
Mammary Glands, Animal/metabolism , Mannheimia haemolytica/physiology , Mastitis/metabolism , Pasteurellaceae Infections/metabolism , Proteome/analysis , Sheep Diseases/metabolism , Animals , Biomarkers/analysis , Biomarkers/blood , Biomarkers/metabolism , Blood Proteins/analysis , Blood Proteins/metabolism , Disease Models, Animal , Female , Lactation/metabolism , Mammary Glands, Animal/microbiology , Mammary Glands, Animal/pathology , Mastitis/blood , Mastitis/pathology , Mastitis/veterinary , Milk/chemistry , Milk/metabolism , Milk Proteins/analysis , Milk Proteins/metabolism , Pasteurellaceae Infections/blood , Pasteurellaceae Infections/pathology , Pasteurellaceae Infections/veterinary , Proteome/metabolism , Proteomics , Sheep , Sheep Diseases/blood , Sheep Diseases/pathologyABSTRACT
Infectious coryza (IC) is often a curse for poultry farmers when it occurs concurrently with several pathogens causing swollen head syndrome. The disease is caused by Avibacterium paragallinarum, which inflicts initial damage to the nasal and respiratory epithelium. This facilitates the progression of disease pathology across the nasal cavity, thereby providing a platform for multiplication of opportunistic microbes. In this study, we attempted to investigate the early entrance and migration pattern of A. paragallinarum in chicken and Japanese quail following experimental infection, by employing an in-house developed polyclonal antiserum against this pathogen. Antigenic-specificity of the raised antiserum was subsequently evaluated through immune-dot blot techniques and counter-current immunoelectrophoresis (CIE). The resultant antiserum characterized the antigen localization within formalin-fixed and partially decalcified nasal tissue sections though immunohistochemistry (IHC). Japanese quail showed prominent localization of the bacterial antigen at 12â h post-infection in anterior turbinates. However, the chicken exhibited a higher level of the bacterial pathogen with intense immuno-reactivity at 24 and 48â h post-inoculation. The decline in immunostaining intensity in the nasal tissue of chicken as well as Japanese quail by 72â h post-infection signifies either an attempt to resolve the infection by the resident immune cells across the nasal passage of the host, or its dissipation by certain inherent innate immune factors present across the nasal passage that are still unknown to us. In the present study, we used a moderately virulent pathogen (A. paragallinarum) that inflicted a mild to moderate degree of damage to histo-architecture of the nasal passage and provided a discernible migratory pattern with fewer alterations, along with provision toward unravelling basics of the immuno-pathogenetic mechanism. This knowledge will support efforts towards the development of a future mucosal nasal vaccine in birds affected with IC.
Subject(s)
Chickens , Coturnix , Pasteurellaceae Infections/veterinary , Pasteurellaceae/immunology , Poultry Diseases/microbiology , Animals , Female , Immunohistochemistry/veterinary , Male , Nasal Cavity/microbiology , Nasal Cavity/pathology , Pasteurellaceae/physiology , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Poultry Diseases/pathology , Rabbits , Random Allocation , Turbinates/microbiology , Turbinates/pathologyABSTRACT
A Japanese Black calf became dehydrated on the first day of life and died on the third day. Gross examination revealed a large amount of yellowish-brown serous fluid in the abdominal cavity and whitish-yellow fibrin in the serosa of the abdominal organs. Patchy red spots were observed throughout the peritoneum, and the outer membrane of the umbilical arteries was dark red. Bacteriologically, Mannheimia haemolytica serotype 2 was isolated from the umbilical arteries and vein, liver, and kidney. Histopathology revealed inflammation with M. haemolytica serotype 2 in the outer membrane of the umbilical arteries and in the serosa of the bladder and intestinal tract. This is the first case of bovine peritonitis with histopathologic and immunohistochemical identification of M. haemolytica.
Subject(s)
Cattle Diseases/microbiology , Mannheimia haemolytica/classification , Pasteurellaceae Infections/veterinary , Peritonitis/veterinary , Animals , Animals, Newborn , Cattle , Fatal Outcome , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Peritonitis/microbiology , Peritonitis/pathologySubject(s)
Cattle Diseases/pathology , Meningoencephalitis/veterinary , Pasteurellaceae Infections/veterinary , Pasteurellaceae/isolation & purification , Animals , Animals, Newborn , Autopsy/veterinary , Cattle , Diagnosis, Differential , Lameness, Animal/etiology , Male , Meningoencephalitis/complications , Meningoencephalitis/pathology , Pasteurellaceae Infections/complications , Pasteurellaceae Infections/pathologyABSTRACT
Infection-related glomerulonephritis (IRGN) develops after various infections. It was previously thought to be caused by Streptococcus species alone but can also be caused by other pathogens. Nephritis-associated plasmin receptor (NAPlr) was discovered as a candidate nephritis-inducing factor in acute post-streptococcal glomerulonephritis. More recently, renal lesions caused by other pathogens were found to be positive for the same molecular marker. We report the case of a 64-year-old man who experienced repeated fever for several months and presented with progressively-deteriorating renal function. He had previously undergone aortic valve replacement. Aggregatibacter actinomycetemcomitans, a component of the oral flora, was detected in a blood culture. Renal biopsy showed diffuse proliferative glomerulonephritis. Immunofluorescence staining of the kidney specimen was positive for immunoglobulins, complements, and NAPlr. The patient was diagnosed with infectious endocarditis and IRGN. Six weeks of intravenous antibiotic therapy improved the patient's clinical condition and kidney function. In this case, IRGN was caused by a rare pathogen. This is the first published case to show NAPlr positivity in the glomeruli after systemic infection with the periodontal bacteria, Aggregatibacter actinomycetemcomitans. This case and subsequent research might expand the concept of IRGN, anchored by NAPlr as a key diagnostic biomarker.â©.
Subject(s)
Aggregatibacter actinomycetemcomitans/isolation & purification , Bacteremia/complications , Glomerulonephritis/diagnosis , Pasteurellaceae Infections/complications , Receptors, Peptide/metabolism , Acute Disease , Bacteremia/metabolism , Bacteremia/pathology , Glomerulonephritis/metabolism , Glomerulonephritis/microbiology , Glomerulonephritis/pathology , Humans , Kidney/metabolism , Kidney/microbiology , Kidney/pathology , Kidney Glomerulus/metabolism , Kidney Glomerulus/microbiology , Kidney Glomerulus/pathology , Male , Middle Aged , Pasteurellaceae Infections/metabolism , Pasteurellaceae Infections/pathologyABSTRACT
Periodontal disease, an inflammatory disease, is caused by infection with periodontal pathogens. Long-term periodontal disease increases the risk of oral carcinogenesis. Similar to other peptic cancers, oral carcinogenesis also requires multiple genome instabilities; however, the risk factors related to the accumulation of genome instabilities are poorly understood. Here, we suggested that specific periodontal pathogens may increase the risk of genome instability. Accordingly, we screened several periodontal pathogens based on the ability to induce DNA double-strand breaks (DSBs) in host cells. We found that Aggregatibacter actinomycetemcomitans Y4 infection induced DSB formation in host cells. To assess whether DSB formation induced by infection with A. actinomycetemcomitans occurred through apoptotic chromosome fragmentation, cells were treated with a caspase inhibitor, Z-VAD-FMK. DSB accumulation induced by infection with A. actinomycetemcomitans was observed, even in the presence of Z-VAD-FMK, suggesting that this breakage occurred independently of apoptosis. These results suggested that some periodontal pathogens can increase the risk of genome instabilities in host cells and subsequently increase the risk of carcinogenesis.
Subject(s)
Aggregatibacter actinomycetemcomitans/physiology , Carcinoma, Squamous Cell/genetics , DNA Breaks, Double-Stranded , Genomic Instability , Pasteurellaceae Infections/microbiology , Tongue Neoplasms/genetics , Apoptosis , Carcinoma, Squamous Cell/microbiology , Carcinoma, Squamous Cell/pathology , Cell Line, Tumor , HeLa Cells , Humans , Pasteurellaceae Infections/pathology , Tongue Neoplasms/microbiology , Tongue Neoplasms/pathologyABSTRACT
Recently we demonstrated that co-infection with Avibacterium paragallinarum and Gallibacterium anatis leads to increased severity of clinical signs of infectious coryza in birds. The present study examined the interaction of these two pathogens in chickens by evaluation of histologic lesions in sinus infraorbitalis and nasal turbinates, applying a defined scoring scheme ranging from 0 to 3. Furthermore, for the first time, an in situ hybridization (ISH) technique was applied to detect A. paragallinarum in tissues. The samples were received from vaccinated and nonvaccinated birds that were infected with A. paragallinarum and/or G. anatis. Vaccinated birds were mostly devoid of any histopathologic lesions except a few birds with lesion score 1 at 7 and 14 days postinfection (dpi). Likewise, nonvaccinated birds infected with G. anatis only did not present microscopic changes in the sinus infraorbitalis, except in a single bird at 7 dpi. Interestingly, median lesion scores caused by G. anatis infection were significantly higher in the nasal turbinates of infected birds than in negative control at 7 and 14 dpi. The most prominent histologic changes were recorded from sinus infraorbitalis and nasal turbinates of nonvaccinated birds that were infected either with A. paragallinarum only or together with G. anatis. ISH demonstrated positive signals for A. paragallinarum in exudates present in the lumen or attached to the epithelial layer of investigated tissues. Such signals were mainly detected in tissues from birds with the highest histopathologic lesion scores.
Subject(s)
Chickens , Coinfection/veterinary , Haemophilus Infections/veterinary , Pasteurellaceae Infections/veterinary , Poultry Diseases/pathology , Animals , Coinfection/microbiology , Coinfection/pathology , Haemophilus Infections/microbiology , Haemophilus Infections/pathology , Haemophilus paragallinarum/physiology , In Situ Hybridization , Paranasal Sinuses/microbiology , Pasteurellaceae/physiology , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Poultry Diseases/microbiology , Turbinates/microbiology , Vaccination/veterinarySubject(s)
Aggregatibacter aphrophilus , Endocarditis, Bacterial/diagnosis , Endocarditis, Bacterial/microbiology , Mitral Valve Annuloplasty , Mitral Valve/microbiology , Pasteurellaceae Infections/diagnosis , Pasteurellaceae Infections/microbiology , Aggregatibacter aphrophilus/genetics , Aggregatibacter aphrophilus/isolation & purification , DNA, Bacterial/genetics , Endocarditis, Bacterial/pathology , Endocarditis, Bacterial/surgery , Humans , Male , Middle Aged , Mitral Valve/pathology , Mitral Valve/surgery , Pasteurellaceae Infections/pathology , Pasteurellaceae Infections/surgery , Polymerase Chain Reaction , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Taiwan , Treatment OutcomeABSTRACT
Gingival epithelial cells form a physiological barrier against bacterial invasion. Excessive bacterial invasion destroys the attachment between the tooth surface and the epithelium, resulting in periodontitis. Integrins play a significant role in cell attachment; therefore, we hypothesized that bacterial infection might decrease the expressions of these integrins in gingival epithelial cells, resulting in reduced cell adhesion. Immortalized human gingival epithelial cells were co-cultured with Aggregatibacter actinomycetemcomitans Y4 (Aa Y4), and the gene expression levels of IL-8, proliferating cell nuclear antigen (PCNA), and integrins (α2, α3, α5, ß4, and ß6) were measured using quantitative reverse transcription polymerase chain reaction. Expression of PCNA and integrins, except integrin α5, was significantly downregulated, while expression of IL-8 and integrin α5 was significantly upregulated in the cells co-cultured with Aa Y4. The number of adherent cells significantly decreased when co-cultured with Aa Y4, as determined using cell adhesion assays. In the cells co-cultured with Aa Y4 and an integrin α5 neutralizing antibody, there was no effect on the expression of IL-8 and PCNA, while the expressions of integrins α2, α3, ß4, and ß6, and the number of adherent cells did not decrease. The number of invading bacteria in the cells was reduced in the presence of the antibody and increased in the presence of TLR2/4 inhibitor. Therefore, integrin α5 might be involved in Aa Y4 invasion into gingival epithelial cells, and the resulting signal transduction cascade reduces cell adhesion by decreasing the expression of integrins, while the TLR2/4 signaling cascade regulates IL-8 expression.
Subject(s)
Aggregatibacter actinomycetemcomitans/metabolism , Epithelial Cells/metabolism , Gene Expression Regulation , Gingiva/metabolism , Integrin alpha Chains/metabolism , Pasteurellaceae Infections/metabolism , Adult , Cell Adhesion , Epithelial Cells/microbiology , Epithelial Cells/pathology , Gingiva/microbiology , Gingiva/pathology , Humans , Male , Pasteurellaceae Infections/pathologyABSTRACT
Avibacterium paragallinarum and Gallibacterium anatis are recognized bacterial pathogens both infecting the respiratory tract of chickens. The present study investigated outcomes of their coinfection by elucidating clinical signs, pathologic lesions, and bacteriologic findings. Additionally, the efficacy of a commercially available vaccine to prevent diseases caused by A. paragallinarum and G. anatis was evaluated. Birds inoculated with G. anatis alone did not present any clinical signs and gross pathologic lesions in the respiratory tract. However, clinical signs of infectious coryza were reproduced in nonvaccinated birds that were challenged with A. paragallinarum alone or together with G. anatis . Such clinical signs were more severe in the coinfected group, including the death of four birds. Some of the birds that were vaccinated and challenged showed mild clinical signs at 7 days postinfection (dpi). Inflammation of sinus infraorbitalis was the most prominent gross pathologic lesion found in the respiratory tract of nonvaccinated birds inoculated either with A. paragallinarum and G. anatis or A. paragallinarum alone. In the reproductive tract, hemorrhagic follicles were observed in nonvaccinated birds that were infected either with G. anatis alone or together with A. paragallinarum . In vaccinated birds, no gross pathologic lesions were found except in one bird that was coinfected with both the pathogens characterized by mucoid tracheitis. Bacteriologic investigations revealed that multiplication of G. anatis at 7 dpi was supported by the coinfection with A. paragallinarum . Altogether, it can be concluded that simultaneous infection of A. paragallinarum and G. anatis can increase the severities of disease conditions in chickens. In such a scenario, vaccination appears to be an effective tool for prevention of the disease, as protection was conferred based on clinical, pathologic, bacteriologic, and serologic data.
Subject(s)
Bacterial Vaccines/administration & dosage , Coinfection/microbiology , Haemophilus Infections/prevention & control , Haemophilus paragallinarum/immunology , Pasteurellaceae Infections/prevention & control , Pasteurellaceae/immunology , Poultry Diseases/prevention & control , Animals , Bacterial Vaccines/immunology , Chickens , Coinfection/pathology , Coinfection/prevention & control , Haemophilus Infections/microbiology , Haemophilus Infections/pathology , Haemophilus paragallinarum/genetics , Haemophilus paragallinarum/physiology , Pasteurellaceae/genetics , Pasteurellaceae/physiology , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Poultry Diseases/microbiology , Poultry Diseases/pathology , Specific Pathogen-Free Organisms , VaccinationABSTRACT
Bovine respiratory syncytial virus (BRSV) and Histophilus somni synergize to cause respiratory disease in cattle. These pathogens cause enhanced disease during dual-infection and an IgE response to antigens of H. somni in dual-infected but not singly infected calves. Vaccines containing whole inactivated BRSV or H. somni have been associated with IgE responses A vaccine strategy that avoids stimulation of IgE antibodies would provide superior protection from dual infection. We hypothesized that a subunit vaccine consisting of the nucleoprotein (NP) from BRSV and the recombinant antigen IbpA DR2 (a surface antigen of H. somni with two toxic fic motifs) in Quil A adjuvant would elicit protection without disease enhancement. Three groups of calves were vaccinated twice with either: Formalin inactivated BRSV (FI) plus Somnivac®, NP & IbpA DR2 plus Quil A or Quil A alone, followed by BRSV and H. somni challenge. Clinical scores and antibody levels (to whole pathogens and to the subunits) were evaluated. Lungs were examined at necropsy on day 23 after infection. Clinical scores were significantly greatest for the FI & Somnivac® group and both clinical scores and lung pathology were lowest for the subunit group. All calves shed BRSV in nasal secretions. FI & Somnivac® induced IgE antibodies to H. somni and BRSV, but not to NP or DR2. The subunit vaccine did not induce an IgE antibody response to IbpA DR2 antigen and induced little IgE to H. somni. It did not induce an IgG antibody response to BRSV and H. somni, but stimulated production of IgG antibodies against the subunits. In summary, the subunit vaccine, consisting of the BRSV NP and H. somni IbpA DR2 in Quil A, protected against severe clinical signs and decreased lung pathology but did not prevent viral shedding. Importantly it prevented synergistic disease expression in response to dual infection.
Subject(s)
Bacterial Vaccines/immunology , Pasteurellaceae Infections/veterinary , Pasteurellaceae/immunology , Respiratory Syncytial Virus Infections/veterinary , Respiratory Syncytial Viruses/immunology , Viral Vaccines/immunology , Adjuvants, Immunologic/administration & dosage , Animals , Antibody Formation , Bacterial Vaccines/administration & dosage , Bacterial Vaccines/genetics , Cattle , Lung/pathology , Pasteurellaceae Infections/pathology , Pasteurellaceae Infections/prevention & control , Quillaja Saponins/administration & dosage , Respiratory Syncytial Virus Infections/prevention & control , Severity of Illness Index , Treatment Outcome , Vaccines, Subunit/administration & dosage , Vaccines, Subunit/genetics , Vaccines, Subunit/immunology , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/genetics , Vaccines, Synthetic/immunology , Viral Vaccines/administration & dosage , Viral Vaccines/geneticsABSTRACT
Histophilus somni is a pathogenic gram-negative bacterium responsible for pneumonia and septicemia in cattle. Sequelae include infectious thrombotic meningoencephalitis (ITME), myocarditis, arthritis, and abortion. These syndromes are associated with widespread vasculitis and thrombosis, implicating a role for endothelium in pathogenesis. Histopathologic and immunohistochemical investigation of 10 natural cases of bovine H. somni myocarditis and 1 case of ITME revealed intravascular H. somni in large biofilm-like aggregates adherent to the luminal surface of microvascular endothelium. Ultrastructurally, bacterial communities were extracellular and closely associated with degenerating or contracted endothelial cells. Histophilus somni was identified by bacterial culture and/or immunohistochemistry. Western blots of the bacterial isolates revealed that they expressed the immunodominant protective 40 kDa OMP and immunoglobulin-binding protein A (IbpA) antigens. The latter is a large surface antigen and shed fibrillar antigen with multiple domains. The cytotoxic DR2Fic domain of IbpA was conserved as demonstrated by polymerase chain reaction. Treatment of endothelial cells in vitro with IbpA in crude culture supernatants or purified recombinant GST-IbpA DR2Fic (rDR2) cytotoxin induced retraction of cultured bovine brain microvascular endothelial cells. By contrast, no retraction of bovine endothelium was induced by mutant rDR2H/A with an inactive Fic motif or by a GST control, indicating that the cytotoxic DR2Fic motif plays an important role in endothelial cell retraction in vasculitis. The formation of biofilm-like aggregates by H. somni on bovine microvascular endothelium may be fundamental to its pathogenesis in heart and brain.