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Frontiers in Cellular and Infection... 2022Respiratory infections seriously affect the swine industry worldwide. Co-infections of two vital pathogenic bacteria () and (), colonizing the respiratory tract often...
Respiratory infections seriously affect the swine industry worldwide. Co-infections of two vital pathogenic bacteria () and (), colonizing the respiratory tract often occurs in veterinary clinical practice. Moreover, our previous research found that and can form biofilm . The formation of a mixed biofilm not only causes persistent infections, but also increases the multiple drug resistance of bacteria, which brings difficulties to disease prevention and control. However, the methods for detecting and in co-infection and biofilm are immature. Therefore, in this study, primers and probes were designed based on the conservative sequence of gene and IVA gene. Then, a TaqMan duplex real-time PCR method for simultaneous detection of and was successfully established optimizing the reaction system and conditions. The specificity analysis results showed that this TaqMan real-time PCR method had strong specificity and high reliability. The sensitivity test results showed that the minimum detection concentration of and recombinant plasmid was 10 copies/μL, which is 100 times more sensitive than conventional PCR methods. The amplification efficiencies of and were 95.9% and 104.4% with R value greater than 0.995, respectively. The slopes of the calibration curves of absolute cell abundance of and were 1.02 and 1.09, respectively. The assays were applied to cultivated mixed biofilms and approximately 10 CFUs per biofilm were quantified when 10 CFUs planktonic bacteria of either or were added to biofilms. In summary, this study developed a TaqMan real-time PCR assay for specific, accurate quantification of or in mixed biofilms, which may help for the detection, prevention and control of diseases caused by a bacterial mixed infection involving and .
Topics: Actinobacillus Infections; Actinobacillus pleuropneumoniae; Animals; Biofilms; Coinfection; Reproducibility of Results; Streptococcus suis; Swine; Swine Diseases
PubMed: 35992166
DOI: 10.3389/fcimb.2022.898412 -
BMC Veterinary Research Dec 2023Accurate measurement of disease associated with endemic bacterial agents in pig populations is challenging due to their commensal ecology, the lack of disease-specific...
Detection and disease diagnosis trends (2017-2022) for Streptococcus suis, Glaesserella parasuis, Mycoplasma hyorhinis, Actinobacillus suis and Mycoplasma hyosynoviae at Iowa State University Veterinary Diagnostic Laboratory.
BACKGROUND
Accurate measurement of disease associated with endemic bacterial agents in pig populations is challenging due to their commensal ecology, the lack of disease-specific antemortem diagnostic tests, and the polymicrobial nature of swine diagnostic cases. The main objective of this retrospective study was to estimate temporal patterns of agent detection and disease diagnosis for five endemic bacteria that can cause systemic disease in porcine tissue specimens submitted to the Iowa State University Veterinary Diagnostic Laboratory (ISU VDL) from 2017 to 2022. The study also explored the diagnostic value of specific tissue specimens for disease diagnosis, estimated the frequency of polymicrobial diagnosis, and evaluated the association between phase of pig production and disease diagnosis.
RESULTS
S. suis and G. parasuis bronchopneumonia increased on average 6 and 4.3%, while S. suis endocarditis increased by 23% per year, respectively. M. hyorhinis and A. suis associated serositis increased yearly by 4.2 and 12.8%, respectively. A significant upward trend in M. hyorhinis arthritis cases was also observed. In contrast, M. hyosynoviae arthritis cases decreased by 33% average/year. Investigation into the diagnostic value of tissues showed that lungs were the most frequently submitted sample, However, the use of lung for systemic disease diagnosis requires caution due to the commensal nature of these agents in the respiratory system, compared to systemic sites that diagnosticians typically target. This study also explored associations between phase of production and specific diseases caused by each agent, showcasing the role of S. suis arthritis in suckling pigs, meningitis in early nursery and endocarditis in growing pigs, and the role of G. parasuis, A. suis, M. hyorhinis and M. hyosynoviae disease mainly in post-weaning phases. Finally, this study highlighted the high frequency of co-detection and -disease diagnosis with other infectious etiologies, such as PRRSV and IAV, demonstrating that to minimize the health impact of these endemic bacterial agents it is imperative to establish effective viral control programs.
CONCLUSIONS
Results from this retrospective study demonstrated significant increases in disease diagnosis for S. suis, G. parasuis, M. hyorhinis, and A. suis, and a significant decrease in detection and disease diagnosis of M. hyosynoviae. High frequencies of interactions between these endemic agents and with viral pathogens was also demonstrated. Consequently, improved control programs are needed to mitigate the adverse effect of these endemic bacterial agents on swine health and wellbeing. This includes improving diagnostic procedures, developing more effective vaccine products, fine-tuning antimicrobial approaches, and managing viral co-infections.
Topics: Humans; Swine; Animals; Mycoplasma hyorhinis; Mycoplasma hyosynoviae; Streptococcus suis; Actinobacillus suis; Mycoplasma Infections; Iowa; Retrospective Studies; Universities; Swine Diseases; Arthritis; Endocarditis
PubMed: 38087358
DOI: 10.1186/s12917-023-03807-w -
The Journal of Veterinary Medical... May 2022The species of the genus Actinobacillus have so far been associated with specific animal hosts, and A. suis sensu stricto, an opportunistic pathogen of swine, is rarely...
The species of the genus Actinobacillus have so far been associated with specific animal hosts, and A. suis sensu stricto, an opportunistic pathogen of swine, is rarely isolated from ruminants. We describe here the isolation of A. suis sensu stricto from a newborn calf that died on a dairy farm in Japan. Identification of the isolate was performed by phenotypic and genotypic characterization, with the latter consisting of nucleotide sequence analyses of the 16S rRNA gene plus three housekeeping genes, rpoB, infB and recN.
Topics: Actinobacillus; Actinobacillus Infections; Actinobacillus suis; Animals; Phylogeny; RNA, Ribosomal, 16S; Swine; Swine Diseases
PubMed: 35387957
DOI: 10.1292/jvms.22-0044 -
Animals : An Open Access Journal From... Oct 2023( HPS) is a prominent pathogenic bacterium in pig production. Its infection leads to widespread fibrinous inflammation in various pig tissues and organs, often in...
( HPS) is a prominent pathogenic bacterium in pig production. Its infection leads to widespread fibrinous inflammation in various pig tissues and organs, often in conjunction with various respiratory virus infections, and leads to substantial economic losses in the pig industry. Therefore, the rapid diagnosis of this pathogen is of utmost importance. In this study, we used recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR) technology to establish a convenient detection and analysis system for that is fast to detect, easy to implement, and accurate to analyze, known as RPA-CRISPR/Cas12a analysis. The process from sample to results can be completed within 1 h with high sensitivity (0.163 pg/μL of DNA template, < 0.05), which is 10 -fold higher than the common PCR method. The specificity test results show that the RPA-CRISPR/Cas12a analysis of did not react with other common pig pathogens, including type II and IX, , , , , and ( < 0.0001). The RPA-CRISPR/Cas12a assay was applied to 15 serotypes of clinical samples through crude extraction of nucleic acid by boiling method, and all of the samples were successfully identified. It greatly reduces the time and cost of nucleic acid extraction. Moreover, the method allows results to be visualized with blue light. The accurate and convenient detection method could be incorporated into a portable format as point-of-care (POC) diagnostics detection for at the field level.
PubMed: 37958075
DOI: 10.3390/ani13213317 -
Journal of Applied Microbiology Nov 2023Swine respiratory disease (SRD) is a major disease complex in pigs that causes severe economic losses. SRD is associated with several intrinsic and extrinsic factors...
AIMS
Swine respiratory disease (SRD) is a major disease complex in pigs that causes severe economic losses. SRD is associated with several intrinsic and extrinsic factors such as host health status, viruses, bacteria, and environmental factors. Particularly, it is known that many pathogens are associated with SRD to date, but most of the test to detect those pathogens can be normally investigated only one pathogen while taking time and labor. Therefore, it is desirable to develop rapidly and efficiently detectable methods those pathogens to minimize the damage caused by SRD.
METHODS AND RESULTS
We designed a multiplex real-time RT-PCR (RT-qPCR) system to diagnose simultaneously 16 pathogens, including nine viruses and seven bacteria associated with SRD, on the basis of single qPCR and RT-qPCR assays reported in previous studies. Multiplex RT-qPCR system we designed had the same ability to single RT-qPCR without significant differences in detection sensitivity for all target pathogens at minimum to maximum genomic levels. Moreover, the primers and probes used in this system had highly specificity because the sets had not been detected pathogens other than the target and its taxonomically related pathogens. Furthermore, our data demonstrated that this system would be useful to detect a causative pathogen in the diagnosis using oral fluid from healthy pigs and lung tissue from pigs with respiratory disorders collected in the field.
CONCLUSIONS
The rapid detection of infected animals from the herd using our system will contribute to infection control and prompt treatment in the field.
Topics: Animals; Swine; Reverse Transcriptase Polymerase Chain Reaction; Swine Diseases; Viruses; Lung; Multiplex Polymerase Chain Reaction; Bacteria
PubMed: 37951290
DOI: 10.1093/jambio/lxad263 -
Microorganisms Sep 2023Antimicrobial resistance is an important health issue in human and veterinary medicine. The aim of this study was to monitor the antimicrobial resistance of three of the...
Antimicrobial resistance is an important health issue in human and veterinary medicine. The aim of this study was to monitor the antimicrobial resistance of three of the most important bacteria involved in porcine respiratory disease. A total of 465 isolates were tested during the 2017-2022 period for antimicrobial susceptibility for (n = 137), (n = 207), and (n = 121) by disk diffusion method. The results were interpreted by CLSI breakpoints, where available. High rates of susceptibility (from 90 to >99%) were observed for cefquinome, ceftiofur, amoxicillin + clavulanic acid, amoxicillin, penicillin, ampicillin, florfenicol, enrofloxacin, marbofloxacin, and trimethoprim-sulfamethoxazole. isolates showed high resistance to streptomycin (77%), gentamycin (45%), tilmicosin (39%) erythromycin (33%), oxytetracycline (19%), and tetracycline (18%). For , the highest rates of resistance were observed for streptomycin (98%), tetracycline (75%), oxytetracycline (72%), doxycycline (52%), and erythromycin (51%). presented a high rate of resistance for streptomycin (63%), tilmicosin (29%), oxytetracycline (13%), and tetracycline (14%). Bacteria isolates maintained high susceptibility against antimicrobial agents usually used against the mainly respiratory tract pathogens of swine. Resistance for streptomycin, tetracycline, oxytetracycline, and tilmicosin was high for all the tested pathogens.
PubMed: 37894069
DOI: 10.3390/microorganisms11102410 -
Antibiotics (Basel, Switzerland) Aug 2023Porcine respiratory disease complex (PRDC) has been a major animal health, welfare, and economic problem in Hungary; therefore, great emphasis should be put on both the...
Porcine respiratory disease complex (PRDC) has been a major animal health, welfare, and economic problem in Hungary; therefore, great emphasis should be put on both the prevention and control of this complex disease. As antibacterial agents are effective tools for control, antibiotic susceptibility testing is indispensable for the proper implementation of antibacterial therapy and to prevent the spread of resistance. The best method for this is to determine the minimum inhibitory concentration (MIC) by the broth microdilution method. In our study, we measured the MIC values of 164 , 65 , and 118 isolates isolated from clinical cases against the following antibacterial agents: amoxicillin, ceftiofur, cefquinome, oxytetracycline, doxycycline, tylosin, tilmicosin, tylvalosin, tulathromycin, lincomycin, tiamulin, florfenicol, colistin, enrofloxacin, and sulfamethoxazole-trimethoprim. Outstanding efficacy against isolates was observed with ceftiofur (100%) and tulathromycin (100%), while high levels of resistance were observed against cefquinome (92.7%) and sulfamethoxazole-trimethoprim (90.8%). Ceftiofur (98.4%), enrofloxacin (100%), florfenicol (100%), and tulathromycin (100%) were found to be highly effective against isolates, while 100% resistance was detected against the sulfamethoxazole-trimethoprim combination. For the isolates, only ceftiofur (100%) was not found to be resistant, while the highest rate of resistance was observed against the sulfamethoxazole-trimethoprim combination (94.3%). An increasing number of studies report multi-resistant strains of all three pathogens, making their monitoring a high priority for animal and public health.
PubMed: 37627719
DOI: 10.3390/antibiotics12081298 -
Journal of Applied Microbiology Aug 2023To survey antibiotic susceptibility of bacteria causing cattle and pig respiratory infections in 10 European countries.
AIMS
To survey antibiotic susceptibility of bacteria causing cattle and pig respiratory infections in 10 European countries.
METHODS AND RESULTS
Non-replicate nasopharyngeal/nasal or lung swabs were collected from animals with acute respiratory signs during 2015-2016. Pasteurella multocida, Mannheimia haemolytica, Histophilus somni from cattle (n = 281), and P. multocida, Actinobacillus pleuropneumoniae, Glaesserella parasuis, Bordetella bronchiseptica, and Streptococcus suis from pigs (n = 593) were isolated. MICs were assessed following CLSI standards and interpreted using veterinary breakpoints where available. Histophilus somni isolates were fully antibiotic susceptible. Bovine P. multocida and M. haemolytica were susceptible to all antibiotics, except tetracycline (11.6%-17.6% resistance). Low macrolide and spectinomycin resistance was observed for P. multocida and M. haemolytica (1.3%-8.8%). Similar susceptibility was observed in pigs, where breakpoints are available. Resistance in P. multocida, A. pleuropneumoniae, and S. suis to ceftiofur, enrofloxacin, and florfenicol was absent or <5%. Tetracycline resistance varied from 10.6% to 21.3%, but was 82.4% in S. suis. Overall multidrug-resistance was low. Antibiotic resistance in 2015-2016 remained similar as in 2009-2012.
CONCLUSIONS
Low antibiotic resistance was observed among respiratory tract pathogens, except for tetracycline.
Topics: Cattle; Animals; Swine; Anti-Bacterial Agents; Bacteria; Pasteurella multocida; Respiratory Tract Infections; Tetracycline; Respiratory System; Microbial Sensitivity Tests; Cattle Diseases; Drug Resistance, Bacterial
PubMed: 37391360
DOI: 10.1093/jambio/lxad132 -
Frontiers in Microbiology 2020() and () are primary swine pathogens that have been frequently co-isolated from pigs suffering from severe respiratory disease. The purpose of this study was to...
() and () are primary swine pathogens that have been frequently co-isolated from pigs suffering from severe respiratory disease. The purpose of this study was to investigate the biological impacts of the interactions between and . A single- and dual-species culture model was established via HA9801 (serotype 2) and CVCC265 (serotype 1). The single or mixed biofilms were imaged by confocal laser scanning microscopy. The biomass and viable cells in biofilms were quantified by crystal violet staining and determination of colony-forming units. The antibiotic susceptibility was determined by a microdilution broth method. The differences in gene transcription in pure- or mixed-species biofilms of and was evaluated by quantitative PCR. and formed two-species biofilms when co-cultured . When co-cultured with , biofilm formation by was significantly increased with the absence of NAD that is necessary for the growth of . Moreover, compared with monocultures, the antibiotic resistance of and was both enhanced in the co-culture model. When grown in dual-species biofilms, for , genes associated with virulence factors, including exotoxins and adhesins, were significantly upregulated. For , virulence factor-related genes , , , and were highly induced. These results suggest that the interspecies interactions between and may be cooperative under specific conditions and may play an important role in the disease progression and persistent infection.
PubMed: 32373078
DOI: 10.3389/fmicb.2020.00507 -
Animals : An Open Access Journal From... Apr 2023Porcine Respiratory Diseases Complex (PRDC) is a multifactorial disease that involves several bacterial pathogens, including , , , , and In pigs, the infection may...
Porcine Respiratory Diseases Complex (PRDC) is a multifactorial disease that involves several bacterial pathogens, including , , , , and In pigs, the infection may cause lesions such pleurisy, which can lead to carcass condemnation. Hence, 1015 carcasses were selected from three different commercial pig farms, where the respiratory conditions were evaluated using slaughterhouse pleurisy evaluation system (SPES) and classified into five groups. In total, 106 pleural and lung fragments were collected for qPCR testing to identify the five abovementioned pathogens. A moderate correlation between the severity of the lesions and the presence of (R = 0.38) and (R = 0.28) was observed. Concerning the lung samples, the severity of the lesions was moderately correlated with the presence of (R = 0.43) and (R = 0.35). Moreover, there was a strong correlation between the presence of and in the pleura (R = 0.82). Finally, this approach may be a useful tool to identify and quantify causative agents of PRDC using qPCR, providing a comprehensive evaluation of its relevance, strength, and potential application in the field as a surveillance tool for veterinarians.
PubMed: 37174529
DOI: 10.3390/ani13091493