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Infection and Immunity Jul 1987Outer membrane protein (OMP)-enriched extracts and whole-cell protein preparations of Haemophilus (Actinobacillus) pleuropneumoniae and related organisms were examined... (Comparative Study)
Comparative Study
Outer membrane protein (OMP)-enriched extracts and whole-cell protein preparations of Haemophilus (Actinobacillus) pleuropneumoniae and related organisms were examined by polyacrylamide gel electrophoresis and immunoblotting. Both the OMP-enriched and whole-cell protein profiles of Actinobacillus suis, A. pleuropneumoniae (NAD-independent biovar), A. lignieresii, and Pasteurella haemolytica were very similar to those of H. pleuropneumoniae serotypes 1 to 8. Antisera prepared against H. pleuropneumoniae typically recognized three major OMP antigens with approximate molecular weights of 17,000 (17K), 32K, and 42K in immunoblots of H. pleuropneumoniae serotypes 1 to 8, Actinobacillus spp., and P. haemolytica. Antisera prepared against Actinobacillus spp. and Haemophilus sp. "minor group" also recognized these 17K, 32K, and 42K antigens. Using absorbed sera, we demonstrated that the 17K antigen had an epitope (or epitopes) common to all the gram-negative organisms examined, including Escherichia coli. The 32K and 42K antigens had epitopes common to members of the family Pasteurellaceae but, in the case of the 32K antigen, also contained unique epitopes. These results provide a basis for understanding the lack of specificity of serodiagnostic tests for H. pleuropneumoniae infection and provide another line of evidence for the association of H. pleuropneumoniae with the genus Actinobacillus.
Topics: Actinobacillus; Animals; Antigens, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Proteins; Cross Reactions; Haemophilus; Immunosorbent Techniques; Molecular Weight; Serotyping; Species Specificity; Swine
PubMed: 3298061
DOI: 10.1128/iai.55.7.1626-1634.1987 -
The Journal of Antimicrobial... Feb 2022Building the European Antimicrobial Resistance Surveillance network in Veterinary medicine (EARS-Vet) was proposed to strengthen the European One Health antimicrobial...
BACKGROUND
Building the European Antimicrobial Resistance Surveillance network in Veterinary medicine (EARS-Vet) was proposed to strengthen the European One Health antimicrobial resistance (AMR) surveillance approach.
OBJECTIVES
To define the combinations of animal species/production types/age categories/bacterial species/specimens/antimicrobials to be monitored in EARS-Vet.
METHODS
The EARS-Vet scope was defined by consensus between 26 European experts. Decisions were guided by a survey of the combinations that are relevant and feasible to monitor in diseased animals in 13 European countries (bottom-up approach). Experts also considered the One Health approach and the need for EARS-Vet to complement existing European AMR monitoring systems coordinated by the ECDC and the European Food Safety Authority (EFSA).
RESULTS
EARS-Vet plans to monitor AMR in six animal species [cattle, swine, chickens (broilers and laying hens), turkeys, cats and dogs], for 11 bacterial species (Escherichia coli, Klebsiella pneumoniae, Mannheimia haemolytica, Pasteurella multocida, Actinobacillus pleuropneumoniae, Staphylococcus aureus, Staphylococcus pseudintermedius, Staphylococcus hyicus, Streptococcus uberis, Streptococcus dysgalactiae and Streptococcus suis). Relevant antimicrobials for their treatment were selected (e.g. tetracyclines) and complemented with antimicrobials of more specific public health interest (e.g. carbapenems). Molecular data detecting the presence of ESBLs, AmpC cephalosporinases and methicillin resistance shall be collected too.
CONCLUSIONS
A preliminary EARS-Vet scope was defined, with the potential to fill important AMR monitoring gaps in the animal sector in Europe. It should be reviewed and expanded as the epidemiology of AMR changes, more countries participate and national monitoring capacities improve.
Topics: Animals; Anti-Bacterial Agents; Bacteria; Cats; Cattle; Chickens; Dogs; Drug Resistance, Bacterial; Female; One Health; Swine
PubMed: 35022739
DOI: 10.1093/jac/dkab462 -
Journal of Bacteriology Dec 2012Here, we report the first complete genome sequence of Actinobacillus suis, an important opportunistic pathogen of swine. By comparing the genome sequence of A. suis with...
Here, we report the first complete genome sequence of Actinobacillus suis, an important opportunistic pathogen of swine. By comparing the genome sequence of A. suis with those of other members of the family Pasteurellaceae, we hope to better understand the role of these organisms in health and disease in swine.
Topics: Actinobacillus suis; Animals; DNA, Bacterial; Genome, Bacterial; Molecular Sequence Data; Sequence Analysis, DNA; Serotyping; Swine
PubMed: 23144422
DOI: 10.1128/JB.01633-12 -
Veterinary Research Mar 2011The acute phase protein (APP) response is an early systemic sign of disease, detected as substantial changes in APP serum concentrations and most disease states...
The acute phase protein (APP) response is an early systemic sign of disease, detected as substantial changes in APP serum concentrations and most disease states involving inflammatory reactions give rise to APP responses. To obtain a detailed picture of the general utility of porcine APPs to detect any disease with an inflammatory component seven porcine APPs were analysed in serum sampled at regular intervals in six different experimental challenge groups of pigs, including three bacterial (Actinobacillus pleuropneumoniae, Streptococcus suis, Mycoplasma hyosynoviae), one parasitic (Toxoplasma gondii) and one viral (porcine respiratory and reproductive syndrome virus) infection and one aseptic inflammation. Immunochemical analyses of seven APPs, four positive (C-reactive protein (CRP), haptoglobin (Hp), pig major acute phase protein (pigMAP) and serum amyloid A (SAA)) and three negative (albumin, transthyretin, and apolipoprotein A1 (apoA1)) were performed in the more than 400 serum samples constituting the serum panel. This was followed by advanced statistical treatment of the data using a multi-step procedure which included defining cut-off values and calculating detection probabilities for single APPs and for APP combinations. Combinations of APPs allowed the detection of disease more sensitively than any individual APP and the best three-protein combinations were CRP, apoA1, pigMAP and CRP, apoA1, Hp, respectively, closely followed by the two-protein combinations CRP, pigMAP and apoA1, pigMAP, respectively. For the practical use of such combinations, methodology is described for establishing individual APP threshold values, above which, for any APP in the combination, ongoing infection/inflammation is indicated.
Topics: Actinobacillus Infections; Actinobacillus pleuropneumoniae; Acute-Phase Proteins; Acute-Phase Reaction; Animals; Enzyme-Linked Immunosorbent Assay; Immunodiffusion; Multivariate Analysis; Mycoplasma Infections; Mycoplasma hyosynoviae; Porcine Reproductive and Respiratory Syndrome; Porcine respiratory and reproductive syndrome virus; Streptococcal Infections; Streptococcus suis; Swine; Swine Diseases; Toxoplasma; Toxoplasmosis; Turpentine
PubMed: 21414190
DOI: 10.1186/1297-9716-42-50 -
Validation of reference genes for quantitative real-time PCR (qPCR) analysis of Actinobacillus suis.BMC Research Notes Mar 2015Quantitative real-time PCR is a valuable tool for evaluating bacterial gene expression. However, in order to make best use of this method, endogenous reference genes for...
BACKGROUND
Quantitative real-time PCR is a valuable tool for evaluating bacterial gene expression. However, in order to make best use of this method, endogenous reference genes for expression data normalisation must first be identified by carefully validating the stability of expression under experimental conditions. Therefore, the objective of this study was to validate eight reference genes of the opportunistic swine pathogen, Actinobacillus suis, grown in aerobic cultures with (Epinephrine) or without (Aerobic) epinephrine in the growth medium and in anoxic static cultures (Anoxic), and sampled during exponential and stationary phases.
RESULTS
Using the RefFinder tool, expression data were analysed to determine whether comprehensive stability rankings of selected reference genes varied with experimental design. When comparing Aerobic and Epinephrine cultures by growth phase, pyk and rpoB were both among the most stably expressed genes, but when analysing both growth phases together, only pyk remained in the top three rankings. When comparing Aerobic and Anoxic samples, proS ranked among the most stable genes in exponential and stationary phase data sets as well as in combined rankings. When analysing the Aerobic, Epinephrine, and Anoxic samples together, only gyrA ranked consistently among the top three most stably expressed genes during exponential and stationary growth as well as in combined rankings; the rho gene ranked as least stably expressed gene in this data set.
CONCLUSIONS
Reference gene stability should be carefully assessed with the design of the experiment in mind. In this study, even the commonly used reference gene 16S rRNA demonstrated large variability in stability depending on the conditions studied and how the data were analysed. As previously suggested, the best approach may be to use a geometric mean of multiple genes to normalise qPCR results. As researchers continue to validate reference genes for various organisms in multiple growth conditions and sampling time points, it may be possible to make informed predictions as to which genes may be most suitable to validate for a given experimental design, but in the meantime, the reference genes used to normalise qPCR data should be selected with caution.
Topics: Actinobacillus suis; Aerobiosis; Anaerobiosis; Bacterial Proteins; Culture Media; Epinephrine; Gene Expression; Gene Expression Profiling; Genes, Essential; Pyruvate Kinase; RNA Polymerase II; RNA, Ribosomal, 16S; Real-Time Polymerase Chain Reaction; Reference Standards
PubMed: 25884823
DOI: 10.1186/s13104-015-1045-8 -
Acta Tropica Nov 2018The productivity of pigs in smallholder systems is affected by high disease burden, most of which might not be obvious, with their epidemiology and impact being poorly...
BACKGROUND
The productivity of pigs in smallholder systems is affected by high disease burden, most of which might not be obvious, with their epidemiology and impact being poorly understood. This study estimated the seroprevalence and identified the risk factors of a range of bacterial and viral pathogens of potential economic and public health importance in domestic pigs in Uganda. A total of 522 clinically healthy pigs were randomly selected from 276 pig farms in Masaka (142) and Lira (134) districts of Uganda in 2015.
RESULTS
Overall the highest animal prevalence was found for Streptococcus suis 73.0% (CI95: 67.0-78.3) in Lira and 68.2% (CI95: 62.7-73.4) in Masaka; followed by Porcine circovirus type 2 with 50.8% (CI95: 44.5-57.2) in Lira and 40.7% (CI95: 35.2-46.5) in Masaka and Actinobacillus pleuro-pneumoniae, 25.6% (CI95: 20.4-31.6) in Lira and 20.5% (CI95: 16.2-25.6) in Masaka. Mycoplasma hyopneumonia prevalence was 20.9% (CI95: 16.2-26.6) in Lira and 10.1% (CI95: 7.1-14.1) in Masaka, while Porcine parvovirus was 6.2% (CI95: 4.0-9.7) in Masaka and 3.4% (CI95: 1.7-6.6) in Lira. Less common pathogens were Influenza A, 8.5% (CI95: 5.6-12.8) in Lira and 2.0% (CI95: 0.9-4.5) in Masaka and Porcine Reproductive and Respiratory Syndrome Virus, 1.7% (CI95: 0.7-4.3) in Lira and 1.3% (CI95: 0.5-3.5) in Masaka. Even less common was Rotavirus A with 0.8% (CI95: 0.2-3.0) in Lira and 0.7% (CI95: 0.2-2.5) in Masaka; the same was for Aujeszky virus with 0.4% (CI95: 0.7-2.4) in Lira and 0.0% (CI95: 0.0-0.1) in Masaka. Co-infection with two pathogens was common and there was a significant association of M. hyo and PCV2 co-occurrence (p = 0.016). Multivariate analysis showed that for S. suis the use of disinfectant reduced odds of sero-positivitey (OR = 0.15; p = 0.017) and pigs less than 6 months were more likely to be infected than older pigs (OR = 3.35; p = 0.047). For M. hyo, crossbred pigs had higher odd of infection compared to local breeds (OR = 1.59; p < 0.001).
CONCLUSIONS
The studied pathogens have high prevalences in smallholder pig production systems and might be silent killers, thus affecting productivity and there is a possibility that some pathogens could spread to humans. Given the limited knowledge of veterinary workers and the poor diagnostic capacities and capabilities in these systems, the diseases are potentially usually under-diagnosed. These findings constitute baseline data to measure the impact of future interventions aiming to reduce disease burden in the pig production systems in Uganda.
Topics: Animals; Animals, Domestic; Circoviridae Infections; Circovirus; Disinfectants; Farms; Influenza A virus; Multivariate Analysis; Mycoplasma hyopneumoniae; Odds Ratio; Orthomyxoviridae Infections; Pneumonia of Swine, Mycoplasmal; Porcine Reproductive and Respiratory Syndrome; Porcine respiratory and reproductive syndrome virus; Prevalence; Risk Factors; Seroepidemiologic Studies; Streptococcal Infections; Streptococcus suis; Sus scrofa; Swine; Swine Diseases; Uganda
PubMed: 29949731
DOI: 10.1016/j.actatropica.2018.06.025 -
Journal of Veterinary Diagnostic... Sep 2008By the end of 2004, the Canadian swine population had experienced a severe increase in the incidence of Porcine circovirus-associated disease (PCVAD), a problem that was...
Development and use of a multiplex real-time quantitative polymerase chain reaction assay for detection and differentiation of Porcine circovirus-2 genotypes 2a and 2b in an epidemiological survey.
By the end of 2004, the Canadian swine population had experienced a severe increase in the incidence of Porcine circovirus-associated disease (PCVAD), a problem that was associated with the emergence of a new Porcine circovirus-2 genotype (PCV-2b), previously unrecovered in North America. Thus, it became important to develop a diagnostic tool that could differentiate between the old and new circulating genotypes (PCV-2a and PCV-2b, respectively). Consequently, a multiplex real-time quantitative polymerase chain reaction (mrtqPCR) assay that could sensitively and specifically identify and differentiate PCV-2 genotypes was developed. A retrospective epidemiologic survey that used the mrtqPCR assay was performed to determine if cofactors could affect the risk of PCVAD. From 121 PCV-2-positive cases gathered for this study, 4.13%, 92.56%, and 3.31% were positive for PCV-2a, PCV-2b, and both genotypes, respectively. In a data analysis using univariate logistic regressions, the PCVAD-compatible (PCVAD/c) score was significantly associated with the presence of Porcine reproductive and respiratory syndrome virus (PRRSV), PRRSV viral load, PCV-2 viral load, and PCV-2 immunohistochemistry (IHC) results. Polytomous logistic regression analysis revealed that PCVAD/c score was affected by PCV-2 viral load (P = 0.0161) and IHC (P = 0.0128), but not by the PRRSV variables (P > 0.9), which suggests that mrtqPCR in tissue is a reliable alternative to IHC. Logistic regression analyses revealed that PCV-2 increased the odds ratio of isolating 2 major swine pathogens of the respiratory tract, Actinobacillus pleuropneumoniae and Streptococcus suis serotypes 1/2, 1, 2, 3, 4, and 7, which are serotypes commonly associated with clinical diseases.
Topics: Animals; Circoviridae Infections; Circovirus; DNA Primers; Genotype; Plasmids; Polymerase Chain Reaction; Quebec; Sensitivity and Specificity; Swine; Swine Diseases; Wasting Syndrome
PubMed: 18776085
DOI: 10.1177/104063870802000503 -
Journal of Clinical Microbiology Feb 2002Tiamulin is a pleuromutilin derivative used in veterinary practice for the control and specific therapy of infections in swine. This report summarizes studies to...
Tiamulin is a pleuromutilin derivative used in veterinary practice for the control and specific therapy of infections in swine. This report summarizes studies to establish standardized susceptibility testing methods, interpretive criteria, and reagent details for use in veterinary methods recently developed by the National Committee for Clinical Laboratory Standards (NCCLS) (standards M31-A and M37-A, NCCLS, Wayne, Pa., 1999). A total of 636 fastidious and nonfastidious animal and human pathogens were processed by using media and procedures described by the NCCLS. Tiamulin disk diffusion tests used a 30-microg disk concentration, and the proposed MIC breakpoints corresponding to levels achievable in animal target tissues (lung) were < or =4 microg/ml for susceptibility and > or =32 microg/ml for resistance. Correlate zone diameters for specific nonfastidious species were as follows: for Pasteurella multocida and staphylococci tested on Mueller-Hinton agar, susceptibility at > or =19 mm and resistance at < or =11 mm, and for Actinobacillus suis, Erysipelothrix rhusiopathiae, and Streptococcus suis tested on enriched chocolate Mueller-Hinton agar, susceptibility at > or =16 mm and resistance at < or =8 mm. When Actinobacillus pleuropneumoniae was tested, a susceptibility breakpoint of < or =16 microg/ml (> or =9 mm) was suggested for veterinary fastidious medium broth and enriched chocolate Mueller-Hinton agar. Absolute categorical agreement between NCCLS dilution and disk diffusion test results with these criteria ranged from 90.5 to 96.2%. Tiamulin susceptibility testing methods appear to be accurate in their categorical classification for indicated species, and their availability will allow immediate testing of animal isolates to guide therapy via appropriate levels of dosing and to monitor the development of resistance for agents in this unique class.
Topics: Anti-Bacterial Agents; Bacteria; Bacterial Infections; Diterpenes; Humans; Microbial Sensitivity Tests; Veterinary Medicine
PubMed: 11825957
DOI: 10.1128/JCM.40.2.461-465.2002 -
The Journal of Veterinary Medical... Feb 2020The etiology of Porcine respiratory disease complex is complicated by infections with multiple pathogens, and multiple infections increase the difficulty in identifying...
The etiology of Porcine respiratory disease complex is complicated by infections with multiple pathogens, and multiple infections increase the difficulty in identifying the causal pathogen. In this present study, we developed a detection system of microbes from porcine respiratory by using TaqMan real-time PCR (referred to as Dempo-PCR) to screen a broad range of pathogens associated with porcine respiratory diseases in a single run. We selected 17 porcine respiratory pathogens (Actinobacillus pleuropneumoniae, Boldetella bronchiseptica, Haemophilus parasuis, Pasteurella multocida, Pasteurella multocida toxin, Streptococcus suis, Mycoplasma hyopneumoniae, Mycoplasma hyorhinis, Mycoplasma hyosynovie, porcine circovirus 2, pseudorabies virus, porcine cytomegalovirus, swine influenza A virus, porcine reproductive and respiratory virus US strain, EU strain, porcine respiratory coronavirus and porcine hemagglutinating encephalomyelitis virus) as detection targets and designed novel specific primer-probe sets for seven of them. In sensitivity test by using standard curves from synthesized DNA, all primer-probe sets showed high sensitivity. However, porcine reproductive and respiratory virus is known to have a high frequency of genetic mutations, and the primer and probe sequences will need to be checked at a considerable frequency when performing Dempo-PCR from field samples. A total of 30 lung samples from swine showing respiratory symptoms on six farms were tested by the Dempo-PCR to validate the assay's clinical performance. As the results, 12 pathogens (5 virus and 7 bacteria) were detected and porcine reproductive and respiratory virus US strain, Mycoplasma hyorhinis, Haemophilus parasuis, and porcine cytomegalovirus were detected at high frequency. These results suggest that Dempo-PCR assay can be applied as a screening system with wide detection targets.
Topics: Animals; Japan; Real-Time Polymerase Chain Reaction; Respiratory Tract Diseases; Sensitivity and Specificity; Sequence Analysis, DNA; Swine; Swine Diseases
PubMed: 31866601
DOI: 10.1292/jvms.19-0063 -
Acta Veterinaria Scandinavica Jun 2008The project "Antibiotic resistance in bacteria of animal origin - II" (ARBAO-II) was funded by the European Union (FAIR5-QLK2-2002-01146) for the period 2003-05. The aim...
BACKGROUND
The project "Antibiotic resistance in bacteria of animal origin - II" (ARBAO-II) was funded by the European Union (FAIR5-QLK2-2002-01146) for the period 2003-05. The aim of this project was to establish a program for the continuous monitoring of antimicrobial susceptibility of pathogenic and indicator bacteria from food animals using validated and harmonised methodologies. In this report the first data on the occurrence of antimicrobial resistance among bacteria causing infections in pigs are reported.
METHODS
Susceptibility data from 17,642 isolates of pathogens and indicator bacteria including Actinobacillus pleuropneumoniae, Streptococcus suis and Escherichia coli isolated from pigs were collected from fifteen European countries in 2002-2004.
RESULTS
Data for A. pleuropneumoniae from infected pigs were submitted from five countries. Most of the isolates from Denmark were susceptible to all drugs tested with the exceptions of a low frequency of resistance to tetracycline and trimethoprim - sulphonamide. Data for S. suis were obtained from six countries. In general, a high level of resistance to tetracycline (48.0 - 92.0%) and erythromycin (29.1 - 75.0%) was observed in all countries whereas the level of resistance to ciprofloxacin and penicillin differed between the reporting countries. Isolates from England (and Wales), France and The Netherlands were all susceptible to penicillin. In contrast the proportion of strains resistant to ciprofloxacin ranged from 12.6 to 79.0% (2004) and to penicillin from 8.1 - 13.0% (2004) in Poland and Portugal. Data for E. coli from infected and healthy pigs were obtained from eleven countries. The data reveal a high level of resistance to tetracyclines, streptomycin and ampicillin among infected pigs whereas in healthy pigs the frequency of resistance was lower.
CONCLUSION
Bacterial resistance to some antimicrobials was frequent with different levels of resistance being observed to several antimicrobial agents in different countries. The occurrence of resistance varied distinctly between isolates from healthy and diseased pigs, with the isolates from healthy pigs generally showing a lower level of resistance than those from diseased pigs. The study suggests that the choice of antimicrobials used for the treatment of diseased animals should preferably be based on knowledge of the local pattern of resistance.
Topics: Actinobacillus pleuropneumoniae; Animals; Anti-Bacterial Agents; Bacterial Infections; Drug Resistance, Multiple, Bacterial; Escherichia coli; Europe; Prevalence; Streptococcus suis; Swine; Swine Diseases; Time Factors
PubMed: 18554407
DOI: 10.1186/1751-0147-50-19