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Introduction: Porcine circovirus type 2 (PCV2) and Porcine epidemic diarrhea virus (PEDV) are highly prevalent in Sichuan, significantly affecting the swine industry's development. PCV2, known for its immunosuppressive effects, can compromise pigs' immune systems, while PEDV typically causes diarrhea in piglets, leading to high mortality rates. Despite their impact, recent studies on the epidemiology and genetic diversity of PCV2 and PEDV within Sichuan Province remain limited. Methods: This study examines clinical samples from 352 diarrheal piglets across 63 pig farms in 17 regions of Sichuan Province, revealing positivity rates of 42.33% (149/352) for PCV2 and 50.28% (177/352) for PEDV, with a co-infection rate of 27.56% (97/352). Notably, the highest positivity rates were observed in Ziyang for PCV2 at 61.90% (13/21), and in Meishan for PEDV at 73.81% (31/42), both regions also reported the highest co-infection rates of 47.62%. Results and discussion: Seasonal analysis indicated that PEDV infections peaked during winter, whereas PCV2 showed no significant seasonal trends. Phylogenetic analysis identified 14 PCV2 strains, categorizing 2 as PCV2b (14.29%), 10 as PCV2d (71.43%), and 2 as PCV2e (14.29%). Among the 16 PEDV strains, 2 were classified as G1a (12.5%) and 14 as G2a (87.5%), with PCV2d and PEDV G2a identified as the predominant strains in the region. The study also highlights a high mutation rate at the antigenic sites of both viruses, potentially affecting vaccine efficacy. These findings underscore the need for ongoing surveillance and vaccine development tailored to the prevalent strains to improve control measures within the province. [ABSTRACT FROM AUTHOR]

Porcine Respiratory Disease Complex (PRDC) presents a complex etiopathology that poses challenges for veterinarians. Typically, a primary microorganism acts as the initiating factor, allowing secondary pathogens to exacerbate respiratory complications. Notable initiating viruses include Swine Influenza Virus (SIV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), and Porcine Circovirus Type 2 (PCV2). Mycoplasma hyopneumoniae (Mhp) and Actinobacillus pleuropneumoniae (App) are frequently cited as initiating bacteria. The profound influence of environmental conditions on animal health underscores the necessity for tailored environmental adjustments. Optimal husbandry practices, including improved living conditions, reduced stocking density, and mitigation of stressors, are pivotal at all production stages. Technological innovations for health monitoring should also be considered. Effective immunoprophylaxis for PRDC is constrained due to its multifactorial nature. However, substantial reductions in associated losses can be achieved through vaccinations targeting PRRSV, Mhp, and App, followed by SIV. Vaccination against PCV2, Bordetella bronchiseptica, and Pasteurella multocida is also justified. Treatment of PRDC necessitates a comprehensive approach encompassing pathogen elimination, supportive therapy, and mitigation of detrimental influences on animal health and immunity. Additionally, specialized respiratory support supplements for farm animals are commercially available. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) poses a significant threat to domestic pigs and wild boar (Sus scrofa) populations, with the current epidemiological situation more critical than ever. The disease has spread across five continents, causing devastating losses in the swine industry. Although extensive research efforts are ongoing to develop an effective and safe vaccine, this goal remains difficult to achieve. Among the potential vaccine candidates, live attenuated viruses (LAVs) have emerged as the most promising option due to their ability to provide strong protection against experimental challenges. However, ASF virus (ASFV) is highly diverse, with genetic and phenotypic variations across different isolates, which differ in virulence. This study highlights the limitations of a natural LAV strain (Lv17/WB/Rie1), which showed partial efficacy against a highly virulent and partially heterologous isolate (Arm07; genotype II). However, the LAV's effectiveness was incomplete when tested against a more phylogenetically distant virus (Ken06.Bus; genotype IX). These findings raise concerns about the feasibility of developing a universal vaccine for ASFV in the near future, emphasizing the urgent need to assess the protective scope of LAV candidates across different ASFV isolates to better define their limitations. [ABSTRACT FROM AUTHOR]

Porcine cysticercosis (PC) is an important public health problem, especially in sub-Saharan Africa, but limited information is available on the prevalence of infection in pigs entering the food chain. Existing diagnostic methods vary in accuracy and efficiency; whole carcass dissection is the most reliable method but is labour-intensive and destroys the carcass so can only be used in a research setting. Serological tests offer lower specificity, while meat inspection and lingual examination lack sensitivity, hampering accurate estimates and the removal of infected pigs from the food chain. Here, we provide the first estimates of PC prevalence in abattoirs in Rwanda. We use whole carcass dissection to determine the diagnostic accuracy of a commercial antigen-ELISA to estimate the true prevalence of infection across Rwanda and identify Taenia species affecting local pigs. We carried out a cross-sectional survey in 6 abattoirs across Rwanda (n = 744 pigs), with whole carcass dissection of a subset of 67 pigs. Cysts were detected in 20/67 (30%) of carcasses, with >1000 cysts in 9/20 (45%) of infected pigs. All cysts were identified as Taenia solium by PCR-RFLP, with no cysts of Taenia hydatigena found. The antigen-ELISA showed a sensitivity of 90% (95% CI: 68–99) and specificity of 85% (95% CI: 72–94), when compared to dissection. Using these estimates, the true prevalence was calculated as 25–43% in two abattoirs in south-west Rwanda, and 2–3% in the rest of the country. Fewer than half of infected pigs were detected by tongue palpation and post-mortem veterinary inspection. Our data indicate a high prevalence of PC in Rwandan abattoirs. Tongue palpation and veterinary inspections, as currently carried out, have little impact in removing cyst-infested pigs from the food chain. Additional interventions are needed, such as proper pig husbandry, treatment and vaccination against cysticercosis, health education, improved sanitation and hygiene, and improved processing and cooking of meat. Author summary: Porcine cysticercosis is a tapeworm infection with a complex life cycle involving pigs and humans. This parasite is the cause of over a third of cases of acquired epilepsy in regions of the world where sanitation is poor and free-range pigs are reared, and is a public health and economic challenge to consumers and farmers. Our study aimed to estimate the prevalence of tapeworm infection in slaughtered pigs in Rwanda, in order to support disease control. We found that porcine cysticercosis is widespread across the country with a high prevalence in the south and south-west. Pigs with very high numbers of tapeworm cysts were identified and constitute a public health concern to consumers. We showed that the locally available tests for infection such as tongue palpation and veterinary meat inspection are not efficient at preventing infected pigs from reaching consumers. Control of cysticercosis in Rwanda will require further interventions that support hygienic pig production practices and improved pork processing and cooking, ensure appropriate on-farm treatment to stop transmission of porcine cysticercosis and human taeniasis and increase community awareness and behaviour change. [ABSTRACT FROM AUTHOR]

Background: The widespread practice of delivering antimicrobials through drinking water to livestock leads to considerable variability in exposure levels among animals, raising concerns regarding disease outbreaks and the emergence of antimicrobial resistance. This variability is primarily driven by three pivotal factors: fluctuations in drug concentration within water pipes, variances in drinking behavior among animals, and differences in individual pharmacokinetic parameters. The objective of this study is to develop and evaluate a strategy that tailors medication delivery based on the drinking patterns of pigs, aiming to improve medication distribution without increasing the overall dose of medication. Results: Our results demonstrate that several distribution strategies based on the animals' drinking behavior can effectively increase their overall exposure. These strategies include increasing the exposure of the least exposed animals, raising the average exposure, maximizing the exposure of the majority of the well-exposed animals, or increasing exposure to ensure that a Pharmacokinetics/Pharmacodynamics (PK/PD) criterion reaches a threshold value for a large number of the animals. In summary, constructing an effective distribution strategy for drinking water requires optimizing a specific criterion. The various criteria and methods for optimizing then are detailed. Conclusions: As examples, this article demonstrate that incorporating the drinking behavior into the delivery of amoxicillin results in an increase in the percentage of piglets reaching an AUC/MIC ratio greater than 25h. Specifically, with Pasteurella multocida, the percentage rises from 30 % to at least 60 % , while with Actinobacillus pleuropneumoniae, it increases from 20 % to more than 70 % . [ABSTRACT FROM AUTHOR]

Actinobacillus pleuropneumoniae, a significant respiratory pig pathogen, is causing substantial losses in the global swine industry. The resistance spectrum of A. pleuropneumoniae is expanding, and multidrug resistance is a severe issue. Horizontal gene transfer (HGT) plays a crucial role in the development of the bacterial genome by facilitating the dissemination of resistance determinants. However, the horizontal transfer of resistance genes via A. pleuropneumoniae-derived outer membrane vesicles (OMVs) has not been previously reported. In this study, we used Illumina NovaSeq and PacBio SequeI sequencing platforms to determine the whole genome sequence of A. pleuropneumoniae GD2107, a multidrug-resistant (MDR) isolate from China. We detected a plasmid in the isolate named pGD2107-1; the plasmid was 5,027 bp in size with 7 putative open reading frames (ORF) and included the floR resistance genes. The carriage of resistance genes in A. pleuropneumoniae OMVs was identified using a polymerase chain reaction (PCR) assay, and then we thoroughly evaluated the influence of OMVs on the horizontal transfer of drug-resistant plasmids. The transfer of the plasmid to recipient bacteria via OMVs was confirmed by PCR. In growth competition experiments, all recipients carrying the pGD2107-1 plasmid exhibited a fitness cost compared to the corresponding original recipients. This study revealed that OMVs could mediate interspecific horizontal transfer of the resistance plasmid pGD2107-1 into Escherichia coli recipient strains and significantly enhance the resistance of the transformants. In summary, A. pleuropneumoniae-OMVs play the pivotal role of vectors for dissemination of the floR gene spread and may contribute to more antimicrobial resistance gene transfer in other Enterobacteriaceae. [ABSTRACT FROM AUTHOR]

AbstractActinobacillus pleuropneumoniae infection causes a high mortality rate in porcine animals. Antimicrobial resistance poses global threats to public health. The current study aimed to determine the antimicrobial susceptibilities and probe the resistome of A. pleuropneumoniae in Taiwan. Herein, 133 isolates were retrospectively collected; upon initial screening, 38 samples were subjected to next-generation sequencing (NGS). Over the period 2017–2022, the lowest frequencies of resistant isolates were found for ceftiofur, cephalexin, cephalothin, and enrofloxacin, while the highest frequencies of resistant isolates were found for oxytetracycline, streptomycin, doxycycline, ampicillin, amoxicillin, kanamycin, and florfenicol. Furthermore, most isolates (71.4%) showed multiple drug resistance. NGS-based resistome analysis revealed aminoglycoside- and tetracycline-related genes at the highest prevalence, followed by genes related to beta-lactam, sulfamethoxazole, florphenicol, and macrolide. A plasmid replicon (repUS47) and insertion sequences (IS10R and ISVAp11) were identified in resistant isolates. Notably, the multiple resistance roles of the insertion sequence IS10R were widely proposed in human medicine; however, this is the first time IS10R has been reported in veterinary medicine. Concordance analysis revealed a high consistency of phenotypic and genotypic susceptibility to florphenicol, tilmicosin, doxycycline, and oxytetracycline. The current study reports the antimicrobial characterization of A. pleuropneumoniae for the first time in Taiwan using NGS.

Abstract Tylosin, an antibiotic with a long history in treating respiratory bacterial infections, has unknown effects on the gut microbiota of healthy and infected pigs. The study aimed to investigate the effect of a therapeutic dose of tylosin on swine gut microbiota and explored the relationship between this effect and tylosin pharmacokinetics (PK). We also assessed whether changes in gut microbiota after tylosin administration differ between healthy animals (n = 7) and animals intranasally co-infected (n = 7) with Actinobacillus pleuropneumoniae and Pasteurella multocida. Both groups were intramuscularly administered with tylosin (20 mg/kg). The 16S rRNA gene analyses revealed a significantly lower species richness and diversity, after tylosin treatment, in the infected than the healthy pigs, with infected pigs having lower levels of Bacteroidetes and Firmicutes and higher levels of Proteobacteria. Greater tylosin exposure (greater area under curve (AUC) and maximum plasma concentration (Cmax), and slower elimination (longer terminal half-life, T1/2) were observed in healthy than infected pigs. Relative abundance of Lactobacillus, Oscillibacter, Prevotella, and Sporobacter was positively and significantly correlated with AUC and Cmax, whereas the abundance of Acinetobacter, Alishewanella, and Pseudomonas was positively and significantly correlated with T1/2 and mean residence time (MRT) of tylosin. Our findings, for the first time, demonstrated significant changes in swine gut microbiota after a single therapeutic dose of tylosin was administered, whereas the effect of these changes on tylosin PK was not evident.

Abstract Background Actinobacillus pleuropneumoniae is a serious pathogen in pigs. The abundant application of antibiotics has resulted in the gradual emergence of drugresistant bacteria, which has seriously affected treatment of disease. To aid measures to prevent the emergence and spread of drug-resistant bacteria, herein, the kill rate and mutant selection window (MSW) of danofloxacin (DAN) against A. pleuropneumoniae were evaluated. Methods For the kill rate study, the minimum inhibitory concentration (MIC) was tested using the micro dilution broth method and time-killing curves of DAN against A. pleuropneumoniae grown in tryptic soy broth (TSB) at a series drug concentrations (from 0 to 64 MIC) were constructed. The relationships between the kill rate and drug concentrations were analyzed using a Sigmoid Emax model during different time periods. For the MSW study, the MIC99 (the lowest concentration that inhibited the growth of the bacteria by ≥ 99%) and mutant prevention concentration (MPC) of DAN against A. pleuropneumoniae were measured using the agar plate method. Then, a peristaltic pump infection model was established to simulate the dynamic changes of DAN concentrations in pig lungs. The changes in number and sensitivity of A. pleuropneumoniae were measured. The relationships between pharmacokinetic/pharmacodynamic parameters and the antibacterial effect were analyzed using the Sigmoid Emax model. Results In kill rate study, the MIC of DAN against A. pleuropneumoniae was 0.016 µg/mL. According to the kill rate, DAN exhibited concentration-dependent antibacterial activity against A. pleuropneumoniae. A bactericidal effect was observed when the DAN concentration reached 4–8 MIC. The kill rate increased constantly with the increase in DAN concentration, with a maximum value of 3.23 Log10 colony forming units (CFU)/mL/h during the 0–1 h period. When the drug concentration was in the middle part of the MSW, drugresistant bacteria might be induced. Therefore, the dosage should be avoided to produce a mean value of AUC24h/MIC99 (between 31.29 and 62.59 h. The values of AUC24h/MIC99 to achieve bacteriostatic, bactericidal, and eradication effects were 9.46, 25.14, and > 62.59 h, respectively. Conclusion These kill rate and MSW results will provide valuable guidance for the use of DAN to treat A. pleuropneumoniae infections.

ABSTRACT The objective of this study was to assess the impact of the vehicle of administration and the prandial state of post weaning piglets on the indices of therapeutic efficacy for different broad‐spectrum antibiotic/pathogen combinations. Pharmacokinetic data were retrieved from previous studies, in which we orally administered oxytetracycline (OTC), fosfomycin (FOS), or amoxicillin (AMX) according to the following treatments: dissolved in soft water to fasted or non‐fasted piglets, dissolved in hard water to fasted or non‐fasted piglets, and mixed with feed. Minimum inhibitory concentration (MIC) values for susceptible strains of bacteria causing swine diseases were obtained from the database of European Committee on Antimicrobial Susceptibility Testing (EUCAST) for each antibiotic. Pharmacokinetic/pharmacodynamic (PK/PD) indices of therapeutic efficacy—drug exposure over the dosing interval (fAUC/MIC) for OTC and FOS; time that free drug concentration remains above MIC (%fT>MIC) for AMX—were calculated for each antibiotic/pathogen combination under each treatment. After all OTC and in‐feed FOS and AMX treatments, the indices of therapeutic efficacy were below the target value for all the study microorganisms. When FOS or AMX were delivered dissolved in soft or hard water, the indices were above the target value over which therapeutic efficacy would be expected for Escherichia coli treated with FOS and, Glaesserella parasuis, Pasteurella multocida, and Actinobacillus pleuropneumoniae treated with AMX. The prandial state of piglets showed no influence on the indices of therapeutic efficacy. Pharmacokinetic profiles of broad‐spectrum antibiotics, specifically the ability to achieve target concentrations, may be largely reduced due to drug interactions with components present in feed or water resulting in a discrepancy with PK/PD principles of prudent and responsible use of antibiotics. [ABSTRACT FROM AUTHOR]

Chlamydiae are bacteria that are intriguing and important at the same time. The genus Chlamydia encompasses many species of obligate intracellular organisms: they can multiply only inside the cells of their host organism. Many, perhaps most animals have their own specifically adapted chlamydial species. In humans, the clinically most relevant species is Chlamydia trachomatis, which has particular importance as an agent of sexually transmitted disease. Pigs are the natural host of Chlamydia suis but may also carry Chlamydia abortus and Chlamydia pecorum. C. abortus and possibly C. suis have anthropozoonotic potential, which makes them interesting to human medicine, but all three species bring a substantial burden of disease to pigs. The recent availability of genomic sequence comparisons suggests adaptation of chlamydial species to their respective hosts. In cell biological terms, many aspects of all the species seem similar but non-identical: the bacteria mostly replicate within epithelial cells; they are taken up by the host cell in an endosome that they customize to generate a cytosolic vacuole; they have to evade cellular defences and have to organize nutrient transport to the vacuole; finally, they have to organize their release to be able to infect the next cell or the next host. What appears to be very difficult and challenging to achieve, is in fact a greatly successful style of parasitism. I will here attempt to cover some of the aspects of the infection biology of Chlamydia, from cell biology to immune defence, epidemiology and possibilities of prevention. I will discuss the pig as a host species and the species known to infect pigs but will in particular draw on the more detailed knowledge that we have on species that infect especially humans. [ABSTRACT FROM AUTHOR]

Tylosin, an antibiotic with a long history in treating respiratory bacterial infections, has unknown effects on the gut microbiota of healthy and infected pigs. The study aimed to investigate the effect of a therapeutic dose of tylosin on swine gut microbiota and explored the relationship between this effect and tylosin pharmacokinetics (PK). We also assessed whether changes in gut microbiota after tylosin administration differ between healthy animals (n = 7) and animals intranasally co-infected (n = 7) with Actinobacillus pleuropneumoniae and Pasteurella multocida. Both groups were intramuscularly administered with tylosin (20 mg/kg). The 16S rRNA gene analyses revealed a significantly lower species richness and diversity, after tylosin treatment, in the infected than the healthy pigs, with infected pigs having lower levels of Bacteroidetes and Firmicutes and higher levels of Proteobacteria. Greater tylosin exposure (greater area under curve (AUC) and maximum plasma concentration (Cmax), and slower elimination (longer terminal half-life, T1/2) were observed in healthy than infected pigs. Relative abundance of Lactobacillus, Oscillibacter, Prevotella, and Sporobacter was positively and significantly correlated with AUC and Cmax, whereas the abundance of Acinetobacter, Alishewanella, and Pseudomonas was positively and significantly correlated with T1/2 and mean residence time (MRT) of tylosin. Our findings, for the first time, demonstrated significant changes in swine gut microbiota after a single therapeutic dose of tylosin was administered, whereas the effect of these changes on tylosin PK was not evident. [ABSTRACT FROM AUTHOR]

Simple Summary: Porcine pleuropneumonia, caused by Actinobacillus pleuropneumoniae, is one of the main causes of massive losses in the pig industry, with a high economic impact. This study aimed to identify A. pleuropneumoniae serotypes isolated from pneumonic lesions in dead pigs, naturally infected, in some Piedmontese farms and to describe the macroscopic and microscopic lesions. The identification of circulating serotypes in swine farming is crucial to adopting optimal strategies to control the diseases, such as vaccination plans and biosecurity measures. Porcine pleuropneumonia (PPP) is one of the main causes leading to massive losses in the pig industry, with high economic impacts. Among different etiological agents, Actinobacillus pleuropneumoniae (APP) is responsible for severe fibrinous-necrotizing pleuropneumonia. A total of 19 different APP serotypes are currently recognized. This study aimed to identify APP serotypes isolated from pneumonic lesions in naturally infected and dead pigs in the Piedmont Region and to describe lesions. A total of 107 dead pigs with a suspected PPP diagnosis were included in this study. Lungs were evaluated using gross-pathology scoring systems, histopathology, and APP isolation and serotypes identification by multiplex PCR were conducted. Gross lung lesions were mainly represented by fibrinous pneumonia and pleuropneumonia. APP was isolated in 20/107 (18.7%) samples. PCR indicated APP DNA presence in 53/107 (49.5%) of lung samples. The most observed serotypes were serotype 2 in 24/53 (45.3%) and serotype 6 in 13/53 (24.5%) samples. Moreover, multiplex PCR results suggested a coinfection of different serotypes in five samples. This study emphasizes the importance of an integrated approach, utilizing various techniques, such as gross- and histopathology, and bacteriological culture and PCR, to enhance the diagnosis of APP infections. [ABSTRACT FROM AUTHOR]

Introduction: Actinobacillus pleuropneumoniae (APP) is a serious pathogen that affects the development of livestock breeding. Due to excessive use of antimicrobial drugs, many multidrug-resistant bacteria have emerged and spread, which have threatened the livestock industry. Therefore, we established a peristaltic pump infection model (PPIM) to evaluate the susceptibility change and pharmacokinetic/pharmacodynamic (PK/PD) integration of tulathromycin against APP during the mutant selection window (MSW) for preventing the emergence of mutant-resistant bacteria. Methods: The 99% minimum inhibitory concentration (MIC99) and mutant prevention concentration (MPC) of tulathromycin against APP were measured using the agar-plate method. After the model of dynamic infection had been established based on tulathromycin data in lungs, different dosages were administered to make the drug concentrations located in different parts of the MSW. The population and sensitivity of APP were monitored. Tulathromycin concentrations were measured by high-performance liquid chromatographytandem mass spectrometry. Finally, a sigmoid Emax model was used to analyze the relationships between PK/PD parameters and antibacterial effects. Results and discussion: The values of MIC, MIC99, and MPC of tulathromycin against APP were 2, 1.4, and 44.8 µg/mL, respectively. The PPIM was stable. An elimination effect without regrowth was observed at 5.6 to 44.8 µg/mL (-4.48 to -7.05 Log10 CFU/mL, respectively). The MIC of APP increased 32-fold at 8 MIC99. AUC168 h/MIC99 had the best fit with the antibacterial effect (R2 = 0.9867). The AUC168 h/MIC99 required to achieve bacteriostatic, bactericidal, and clearance effects were 1.80, 87.42, and 198 h, respectively. Our results could provide guidance for the clinical application of tulathromycin to treat APP infection and avoid the generation of drug-resistant bacteria. [ABSTRACT FROM AUTHOR]

Porcine circoviruses (PCVs) are widely distributed in swine herds. PCV2, the significant swine pathogen, causes infections characterized by growth and development disorders, skin lesions, and respiratory distress. PCV3 has been circulating worldwide and can be associated with various clinical signs and disease developments. Wild boars are the main reservoir of these pathogens in wildlife and can create an alarming threat to pig farming. In Russia, three PCV2 genotypes (PCV2a, PCV2b, and PCV2d) were identified in pig farms. Additionally, PCV3 was observed in pig herds during the monitoring studies in the country. However, data considering the circulation of PCVs in herds of wild boars in Russia is scant. For this purpose, we performed PCR assays of the samples from 30 wild boars hunted in the Moscow Region of Russia in 2021–2023. The ratios of wild boars positive for PCV2, PCV3, or coinfected were 50, 10, and 13.3%, respectively. Additionally, we sequenced 15 PCV2 and four PCV3 complete genomes and conducted phylogenetic analysis, which divided PCV2 isolates into two groups: PCV2d and PCV2b. The study showed a high infection rate of PCV2 among wild boars, with PCV2d dominance. Simultaneously, PCV3 also circulates among wild boars. The obtained results can provide a basis for the development of preventive measures to support infection transmission risks between farm and wild animals. [ABSTRACT FROM AUTHOR]

The article focuses on infectious diseases in the context of clinical xenotransplantation, exploring the implications and challenges posed by transferring animal organs into human recipients, with an emphasis on managing infectious risks and ensuring patient safety in this evolving field of medical practice.

Hepatitis E virus (HEV) is a prevalent pathogen responsible for acute viral hepatitis, HEV genotypes 3 and 4 infections causing zoonotic infections. Currently, the nucleotide similarity analysis between humans and pigs for HEV genotype 4 is limited. In this study, stool samples from an HEV‐infected patient who is a pig farmer and from pigs were collected to obtain the near full‐length genome of HEV, phylogenetic trees were constructed for genotyping, and similarity of HEV sequences was analyzed. The results showed that HEV‐RNA was detected in the stool samples from the patient and six pigs (6/30, 20.0%). Both HEV subtype in the patient and pigs was 4b. Additionally, similarity analysis showed that the range was 99.875%–99.944% between the patient and pigs at the nucleotide level. Four isolates of amino acid sequences (ORFs 1–3) from pigs were 100% identical to the patient. Phylogenetic tree and similarity analysis of an additional nine HEV sequences isolated from other patients in this region showed that the HEV sequence from the pig farmer had the closest relationship with the pigs from his farm rather than other sources of infection in this region. This study provides indirect evidences for HEV subtype 4b can be transmitted from pigs to humans at the nucleotide level. Further research is needed to explore the characteristics of different HEV subtypes. [ABSTRACT FROM AUTHOR]

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Actinobacillus pleuropneumoniae is a major bacterial pathogen causing porcine pleuropneumoniae, which is a disease of notable economic impact and high fatality rates among pigs worldwide. It has been reported that 19 distinct serotypes of this bacterium exist. Despite its global prominence, there exists a scarcity of information regarding its prevalence and distribution in South Africa. Thus, this study used laboratory records to investigate the serotype diversity, temporal distribution, and seasonal patterns of A. pleuropneumoniae isolated from porcine samples spanning from 1985 to 2023 within South Africa. Data from laboratory registries of 354 cases, obtained from three veterinary laboratories in South Africa, were analyzed. The data were categorized into two-time frames: term 1, covering 1985 to 2001, and term 2, spanning from 2002 to 2023. The dataset identified 11 different serotypes, with serotype 7 being the most prevalent at 22.7% (n = 62), which was followed by serotype 5 at 13.8% (n = 42). The study highlighted variations in the prevalence of serotypes among diseased animals over a 38-year period. Serotypes 3, 5, 7 and 8 were commonly observed during this time, while serotype 4 was absent from 1985 to 2001, and serotypes 1, 6, and 10 were absent from 2002 to 2023. The distribution of serotypes showed a diverse variation in the age of affected animals, clinical manifestation, and seasonal occurrence. Key findings revealed that serotype 7 was the most prevalent across all seasons with the highest occurrence in winter. Additionally, Gauteng province showed the highest prevalence of various serotypes. The information collected during this study will serve as a baseline for future epidemiological studies as well as inform control strategies. [ABSTRACT FROM AUTHOR]

Background: Actinobacillus pleuropneumoniae is a serious pathogen in pigs. The abundant application of antibiotics has resulted in the gradual emergence of drugresistant bacteria, which has seriously affected treatment of disease. To aid measures to prevent the emergence and spread of drug-resistant bacteria, herein, the kill rate and mutant selection window (MSW) of danofloxacin (DAN) against A. pleuropneumoniae were evaluated. Methods: For the kill rate study, the minimum inhibitory concentration (MIC) was tested using the micro dilution broth method and time-killing curves of DAN against A. pleuropneumoniae grown in tryptic soy broth (TSB) at a series drug concentrations (from 0 to 64 MIC) were constructed. The relationships between the kill rate and drug concentrations were analyzed using a Sigmoid Emax model during different time periods. For the MSW study, the MIC99 (the lowest concentration that inhibited the growth of the bacteria by ≥ 99%) and mutant prevention concentration (MPC) of DAN against A. pleuropneumoniae were measured using the agar plate method. Then, a peristaltic pump infection model was established to simulate the dynamic changes of DAN concentrations in pig lungs. The changes in number and sensitivity of A. pleuropneumoniae were measured. The relationships between pharmacokinetic/pharmacodynamic parameters and the antibacterial effect were analyzed using the Sigmoid Emax model. Results: In kill rate study, the MIC of DAN against A. pleuropneumoniae was 0.016 µg/mL. According to the kill rate, DAN exhibited concentration-dependent antibacterial activity against A. pleuropneumoniae. A bactericidal effect was observed when the DAN concentration reached 4–8 MIC. The kill rate increased constantly with the increase in DAN concentration, with a maximum value of 3.23 Log10 colony forming units (CFU)/mL/h during the 0–1 h period. When the drug concentration was in the middle part of the MSW, drugresistant bacteria might be induced. Therefore, the dosage should be avoided to produce a mean value of AUC24h/MIC99 (between 31.29 and 62.59 h. The values of AUC24h/MIC99 to achieve bacteriostatic, bactericidal, and eradication effects were 9.46, 25.14, and > 62.59 h, respectively. Conclusion: These kill rate and MSW results will provide valuable guidance for the use of DAN to treat A. pleuropneumoniae infections. [ABSTRACT FROM AUTHOR]

It is crucial to discover novel antimicrobial drugs to combat resistance. This study investigated the antibacterial properties of halicin (SU3327), an AI-identified anti-diabetic drug, against 13 kinds of common clinical pathogens of animal origin, including multidrug-resistant strains. Employing minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assessments, halicin demonstrated a broad-spectrum antibacterial effect. Time-killing assays revealed its concentration-dependent bactericidal activity against Escherichia coli ATCC 25922 (E. coli ATCC 25922), Staphylococcus aureus ATCC 29213 (S. aureus ATCC 29213), and Actinobacillus pleuropneumoniae S6 (APP S6) after 4 h of treatment at concentrations above the MIC. Halicin exhibited longer post-antibiotic effects (PAEs) and sub-MIC effects (PA-SMEs) for E. coli 25922, S. aureus 29213, and APP S6 compared to ceftiofur and ciprofloxacin, the commonly used veterinary antimicrobial agents, indicating sustained antibacterial action. Additionally, the results of consecutive passaging experiments over 40 d at sub-inhibitory concentrations showed that bacteria exhibited difficulty in developing resistance to halicin. Toxicology studies confirmed that halicin exhibited low acute toxicity, being non-mutagenic, non-reproductive-toxic, and non-genotoxic. Blood biochemical results suggested that halicin has no significant impact on hematological parameters, liver function, and kidney function. Furthermore, halicin effectively treated respiratory A. pleuropneumoniae infections in murine models. These results underscore the potential of halicin as a new antibacterial agent with applications against clinically relevant pathogens in veterinary medicine. [ABSTRACT FROM AUTHOR]

Porcine circovirus-associated diseases (PCVAD) caused by Porcine Circo Virus-2 (PCV-2) is an emerging viral disease with serious effects on animal health, food security, and the swine industry. In this study, samples were collected from the PCV-2 suspected swine carcasses presented to the Department of Veterinary Pathology, CVAS, Mannuthy, for necropsy. Out of 39 suspected samples, seven were positive for PCV-2 by polymerase chain reaction targeting the ORF-2 gene, which yielded an amplicon size of 481 bp. The blast analysis sequences of the present isolates showed more than 98 percent homology with other parts of India and foreign isolates. The genotypic analysis revealed different PCV-2 genotypes, viz., PCV-2d (57 percent), PCV-2b (29 percent), and PCV-2h (14 percent), and clusters 13, 11, and 18, respectively, for the first time in Kerala. The microscopic examination revealed lymphoid depletion in the spleen, soft palate tonsils, various lymph nodes, ileal Peyer's patches (IPP), and jejunal Peyer's patches (JPP). There were occasional botryoid inclusion bodies in mucosa-associated lymphoid tissues (MALT) with congestion in various lymph nodes. Based on history and clinical signs, gross, histopathological, and PCR results, and sequence data, the presence of PCV-2-associated systemic disease was confirmed in this study. Altogether, these findings are helpful in further understanding the pathogenesis of PCV-2, which would help to evolve better strategies for improved disease control and prevention in pigs. Future investigations on the pathogenesis of these new genetic variants of isolates obtained in the present study are required for a better understanding of the pathogenesis of the disease. [ABSTRACT FROM AUTHOR]

Actinobacillus pleuropneumoniae is the causative agent of pleuropneumonia, an economically important lung disease in pigs. In draft genomes of two Cypriot clinical A. pleuropneumoniae isolates (MIDG3457 and MIDG3459), we previously identified single genomic regions with homology to Mu-like bacteriophage and presented preliminary evidence of active phage. Here, updated Phastest genomic analysis identified two loci in both MIDG3457 and MIDG3459 that were predicted to encode proteins with high homology to, and whose organisation was characteristic of, Mu-like phages. Phylogenetically, the closest matches were with Mannheimia Vb and Glaesserella SuMu phages. Phastest scored the loci as “complete”, indicating they produced active phage. PCR amplification of the Mu-like phage c and tail genes from DNase-treated polyethylene glycol 8000 (PEG)-precipitated supernatants of MIDG3457 and MIDG3459 (grown in either Brain Heart Infusion-NAD or Grace’s Insect Medium-NAD broth) indicated the presence of intact virions. The phages from MIDG3457 and MIDG3459 were named PluMu 3457-1, 3457-2, and PluMu 3459-1 and PluMu 3459-2, respectively. Transmission electron microscopy (TEM) of the PEG-precipitated supernatants of broth-grown MIDG3459 identified virions with icosahedral heads and tails, consistent with other Mu-like phages. We conclude that MIDG3459 produces an active Mu-like phage.

ABSTRACTActinobacillus pleuropneumoniae (APP) is an important pathogen of the porcine respiratory disease complex, which leads to huge economic losses worldwide. We previously demonstrated that Pichia pastoris-producing bovine neutrophil β-defensin-5 (B5) could resist the infection by the bovine intracellular pathogen Mycobacterium bovis. In this study, the roles of synthetic B5 in regulating mucosal innate immune response and protecting against extracellular APP infection were further investigated using a mouse model. Results showed that B5 promoted the production of tumour necrosis factor (TNF)-α, interleukin (IL)-1β, and interferon (IFN)-β in macrophages as well as dendritic cells (DC) and enhanced DC maturation in vitro. Importantly, intranasal B5 was safe and conferred effective protection against APP via reducing the bacterial load in lungs and alleviating pulmonary inflammatory damage. Furthermore, in the early stage of APP infection, we found that intranasal B5 up-regulated the secretion of TNF-α, IL-1β, IL-17, and IL-22; enhanced the rapid recruitment of macrophages, neutrophils, and DC; and facilitated the generation of group 3 innate lymphoid cells in lungs. In addition, B5 activated signalling pathways associated with cellular response to IFN-β and activation of innate immune response in APP-challenged lungs. Collectively, B5 via the intranasal route can effectively ameliorate the immune suppression caused by early APP infection and provide protection against APP. The immunization strategy may be applied to animals or human respiratory bacterial infectious diseases. Our findings highlight the potential importance of B5, enhancing mucosal defence against intracellular bacteria like APP which causes early-phase immune suppression.

African swine fever (ASF), caused by the ASF virus (ASFV), is an acute, severe, and highly contagious infectious disease in domestic pigs and wild boars. Domestic pigs infected with a virulent ASFV strain can have morbidity and mortality rates of up to 100%. The epidemic of ASF has caused serious economic losses to the global pig industry. Currently, there is no safe and effective vaccine or specific drug for treating ASF. Therefore, ASFV still poses a great threat to pig factories. ASFV is a double-stranded DNA virus with a complex icosahedral multilayer structure. The ASFV genome contains 150–170 open reading frames (ORFs) that encode 150–200 proteins. Some ASFV-encoded proteins are involved in virus invasion, genome replication, DNA repair, and virion formation. Some ASFV proteins execute immunomodulatory functions by regulating the host antiviral innate immune response. Accumulating studies have shown that the immunomodulatory functions of ASFV genes are closely related to the virulence and pathogenicity of ASFV isolates. This review summarizes the research advances on ASFV immune evasion mechanisms in African swine fever patients and provides new insights for developing attenuated live vaccine candidates to prevent and control ASF. [ABSTRACT FROM AUTHOR]

Ascaris spp. undergo extensive migration within the body before establishing patent infections in the small intestinal tract of humans and pigs. However, whether larval migration is critical for inducing efficient type 2 responses remains poorly understood. Therefore, we investigated systemic versus local adaptive immune responses along the hepato-tracheal migration of Ascaris suum during primary, single infections in conventionally raised pigs. Neither the initial invasion of gut tissue nor migration through the liver resulted in discernable Th2 cell responses. In contrast, lung-stage larvae elicited a Th2-biased pulmonary response, which declined after the larvae had left the lungs. In the small intestine, we observed an accumulation of Th2 cells upon the arrival of fourth-stage larvae (L4) to the small intestinal lumen. In parallel, we noticed robust and increasing Th1 responses in circulation, migration-affected organs, and draining lymph nodes. Phenotypic analysis of CD4+ T cells specifically recognizing A. suum antigens in the circulation and lung tissue of infected pigs confirmed that the majority of Ascaris-specific T cells produced IL-4 (Th2) and, to a much lesser extent, IL-4/IFN-g (Th2/1 hybrids) or IFN-g alone (Th1). These data demonstrate that lungstage but not the early liver-stage larvae lead to a locally restricted Th2 response. Significant Th2 cell accumulation in the small intestine occurs only when L4 complete the body migration. In addition, Th2 immunity seems to be hampered by the concurrent, nonspecific Th1 bias in growing pigs. Together, the late onset of Th2 immunity at the site of infection and the Th1-biased systemic immunity likely enable the establishment of intestinal infections by sufficiently large L4 stages and pre-adult worms, some of which resist expulsion mechanisms. [ABSTRACT FROM AUTHOR]

Phytochemical studies have shown extracts obtained from the aerial parts of Baccharis trimera (Less.) DC has antioxidant and antimicrobial activities and the potential to treat some diseases. This study investigated the phenolic compounds, antioxidant and antimicrobial activity, and phytochemical potential of B. trimera leaf extract obtained by decoction on ATCC standard bacterial strains and 23 swine clinical isolates. Water was used as an extraction solvent based on the principles of green chemistry and at a low cost. The decoction process resulted in an extract rich in phenolic compounds and a high capacity for scavenging DPPH and ABTS radicals. Phytochemical analysis of aqueous extracts was performed using HPLC-DAD, and high concentrations of chlorogenic, ferulic, caffeic, and cinnamic phenolic acids were found. Antimicrobial activity was observed against gram-negative bacteria. B. trimera aqueous extract may be a promising low-cost agent for prophylactic treatment against swine enteropathogens and contribute to reducing production costs. [ABSTRACT FROM AUTHOR]

This article discusses the development and application of a multiplex real-time PCR assay for the detection of porcine circovirus (PCV) types 2, 3, and 4 in Guangdong province, China. The assay was found to be accurate and reliable, with a high detection limit and no cross-reaction with other porcine viral pathogens. The researchers also conducted an epidemiological investigation and obtained complete genome sequences of PCV2 and PCV3 strains. The study found high infection rates of PCV2 and PCV3 in swine farms in Guangdong Province, with PCV2d identified as the main prevalent genotype. The findings contribute to understanding the epidemiology and genetic variation of PCVs in the region. Additionally, there are other articles referenced that provide valuable information on the prevalence and genetic analysis of PCV types 2 and 3 in Central China, the detection of PCV type 2a and Pasteurella multocida in pigs with respiratory disease, and the occurrence of PCV type 3 without clinical infection signs in Shandong Province. These articles are useful for researchers studying PCV and its impact on pig health. [Extracted from the article]

Actinobacillus pleuropneumoniae (APP) is a major cause of lung infections in pigs. An experimental mouse has the edge over pigs pertaining to the ease of experimental operation, disease study and therapy, abundance of genetic resources, and cost. However, it is a challenge to introduce APP into a mouse lung due to the small respiratory tract of mice and bacterial host tropism. In this study, an effective airborne transmission of APP serovar 1 (APP1) was developed in mice for lung infection. Consequently, APP1 infected BALB/c mice and caused 60% death within three days of infection at the indicated condition. APP1 seemed to enter the lung and, in turn, spread to other organs of the mice over the first 5 days after infection. Accordingly, APP1 damaged the lung as evidenced by its morphological and histological examinations. Furthermore, ampicillin fully protected mice against APP1 as shown by their survival, clinical symptoms, body weight loss, APP1 count, and lung damages. Finally, the virulence of two extra APP strains, APP2 and APP5, in the model was compared based on the survival rate of mice. Collectively, this study successfully established a fast and reliable mouse model of APP which can benefit APP research and therapy. Such a model is a potentially useful model for airway bacterial infections. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome virus (PRRSV) remains one of the major threats to swine industry, resulting in huge economic losses worldwide. Currently, PRRSV has diversified into multiple lineages with characteristics of extensive recombination in China. In this research, three virus strains were isolated and four virus whole genome sequences were generated and analyzed from clinical samples collected in Gansu province of China in 2023. The four virus strains were designated GSTS4-2023, GSLX2-2023, GSFEI2-2023 and GSBY4-2023. Phylogenetic analysis based on ORF5 sequences showed that GSTS4-2023, GSLX2-2023, GSFEI2-2023 and GSBY4-2023 shared 91.7, 91.2, 93.2 and 92.9% homology with NADC30 strain respectively, and belonged to lineage 1 of PRRSV-2. In addition, one amino acid deletion was observed at position 33 in ORF5 of GSTS4-2023, GSLX2-2023 and GSFEI2-2023. Moreover, amino acid alignment of the four strains showed a typical discontinuous 131-amino acid (aa) deletion in NSP2 for NADC30-like virus strains. Recombination analysis revealed that all four strains originated from NADC30 (lineage 1), with their minor parents coming from JXA1-like strains (lineage 8), VR-2332-like strains (lineage5) and QYYZ-like strains (lineage3). Finally, the three isolated virus strains, GSTS4-2023, GSLX2-2023 and GSFEI2-2023 showed relatively low levels of replication in cell culture. Our findings provide important implications for the field epidemiology of PRRSV. [ABSTRACT FROM AUTHOR]

Introduction: Early detection of the virus in the environment or in infected pigs is a critical step to stop African swine fever virus (ASFV) transmission. The p22 protein encoded by ASFV KP177R gene has been shown to have no effect on viral replication and virulence and can serve as a molecular marker for distinguishing field virus strains from future candidate KP177R deletion vaccine strains. Methods: This study established an ASFV detection assay specific for the highly conserved ASFV KP177R gene based on recombinase polymerase amplification (RPA) and the CRISPR/Cas12 reaction system. The KP177R gene served as the initial template for the RPA reaction to generate amplicons, which were recognized by guide RNA to activate the trans-cleavage activity of Cas12a protein, thereby leading to non-specific cleavage of single-stranded DNA as well as corresponding color reaction. The viral detection in this assay could be determined by visualizing the results of fluorescence or lateral flow dipstick (LFD) biotin blotting for color development, and was respectively referred to as fluorescein-labeled RPA-CRISPR/Cas12a and biotin-labeled LFD RPA-CRISPR/Cas12a. The clinical samples were simultaneously subjected to the aforementioned assay, while real-time quantitative PCR (RT-qPCR) was employed as a control for determining the diagnostic concordance rate between both assays. Results: The results showed that fluorescein- and biotin-labeled LFD KP177R RPA-CRISPR/Cas12a assays specifically detected ASFV, did not cross-react with other swine pathogens including PCV2, PEDV, PDCoV, and PRV. The detection assay established in this study had a limit of detection (LOD) of 6.8 copies/mL, and both assays were completed in 30 min. The KP177R RPA-CRISPR/Cas12a assay demonstrated a diagnostic coincidence rate of 100% and a kappa value of 1.000 (p < 0.001), with six out of ten clinical samples testing positive for ASFV using both KP177R RPA-CRISPR/Cas12a and RT-qPCR, while four samples tested negative in both assays. Discussion: The rapid, sensitive and visual detection assay for ASFV developed in this study is suitable for field application in swine farms, particularly for future differentiation of field virus strains from candidate KP177R gene-deleted ASFV vaccines, which may be a valuable screening tool for ASF eradication. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS), an important viral disease of swine caused by PRRS virus (PRRSV) was first confirmed in Nepal in 2013. Since then, the virus has spread throughout the country and has now become endemic affecting the pig production nationally. However, molecular characterization of circulating strains has not been done in Nepal yet. In the present study, serum samples were collected from outbreak areas of different districts of Nepal and samples positive for PRRSV by ELISA were sent to Animal and Plant Health Agency (APHA), United Kingdom for sequence analysis. Out of 35 samples that were sent to APHA, only one sample was found positive by PCR and subjected to sequence analysis based on ORF5, ORF7 and Nsp2. The results from the phylogenetic analysis demonstrated that the PRRSV strain belongs to PRRSV-2 and lineage 8 strain. The sequences from the Nepalese PRRSV strain revealed a high degree of similarity with the strains isolated from India, China and Vietnam, with the closest genetic relatedness to the Indian isolates from 2020 and 2018. This is the first study on molecular characterization of PRRS virus circulating in Nepal. Further studies on strains circulating in Nepal are very essential to understand the virus diversity, its spread and evolution. [ABSTRACT FROM AUTHOR]

African swine fever (ASF) is a highly contagious and lethal viral disease that causes severe hemorrhagic fever in pigs. It keeps spreading around the world, posing a severe socioeconomic risk and endangering biodiversity and domestic food security. ASF first outbroke in China in 2018, and has spread to most provinces nationwide. Genotypes I and II ASF virus (ASFV) as the etiological pathogens have been found in China. In this study, three pairs of specific primers and probes targeting the ASFV B646L gene, F1055L gene, and E183L gene were designed to detect universal, genotype I, and genotype II strains, respectively. A triplex crystal digital PCR (cdPCR) was established on the basis of optimizing various reaction conditions. The assay demonstrated remarkably sensitive with low limits of detection (LODs) of 5.120, 4.218, 4.588 copies/reaction for B646L, F1055L, and E183L gene, respectively; excellent repeatability with 1.24--2.01% intra-assay coefficients of variation (CVs) and 1.32--2.53% inter-assay CVs; good specificity for only detection of genotypes I and II ASFV, without cross-reactivity with PCV2, PRV, SIV, PRRSV, PEDV, FMDV, and CSFV. The triplex cdPCR was used to test 1,275 clinical samples from Guangxi province of China, and the positivity rates were 5.05, 3.22, and 1.02% for genotype I, genotype II, and co-infection of genotypes I and II, respectively. These 1,275 clinical samples were also detected using a reported reference triplex real-time quantitative PCR (qPCR), and the agreements of detection results between these two methods were more than 98.98%. In conclusion, the developed triplex cdPCR could be used as a rapid, sensitive, and accurate method to detect and differentiate genotypes I and II strains of ASFV. [ABSTRACT FROM AUTHOR]

This study aim to explore the application of microdialysis in pharmacokinetic (PK) and pharmacodynamic (PD) integration of cefquinome against Actinobacillus pleuropneumoniae in a porcine experimental lung infection model. The model was established via intratracheal inoculation where average bacterial counts (CFU) in the lungs of infected pigs reached 6.57 log10 CFU/g after 3h. The PK profiles of unbound cefquinome in lung dialysates were determined following intramuscular injection of single doses of 0.125, 0.25, 0.5, 1, 2, and 4mg/kg. Lung dialysate samples were collected using microdialysis at a flow rate of 1.5 μL/min until 24h. The PD studies were conducted over 24h based on 10 intermittent dosing regimens and total daily doses ranged from 0.25 to 4mg/kg and dosage intervals included 12 and 24h. The lung tissue was collected after 24h of treatment and homogenized for bacterial counts. The relationships between PK/PD parameters derived from lung dialysates and drug efficacy were analyzed using an inhibitory sigmoid Emax model. The percentage of time the free drug concentration exceeded the minimum inhibitory concentration (%fT> MIC) was the PK/PD index best describing the antimicrobial activity (R2 = 0.96) in the porcine experimental infection model. The %fT> MIC values required to achieve net bacterial stasis, 1, 2 and 3 log10 CFU/g reductions in the lung were 22.45, 28.86, 37.62, and 56.46%, respectively. Cefquinome exhibited time-dependent characteristics against A. pleuropneumoniae in vivo. These results provide valuable insights into the application of microdialysis in PK/PD integration model studies and optima regimen of cefquinome for the treatment of porcine respiratory diseases caused by A. pleuropneumoniae. [ABSTRACT FROM AUTHOR]

Porcine contagious pleuropneumonia, caused by Actinobacillus pleuropneumoniae, has resulted in significant economic losses to the swine industry. Although antibiotics are commonly employed to control this disease, their widespread use or misuse can lead to the development of antibiotic resistance in A. pleuropneumoniae. Consequently, it is crucial to conduct antimicrobial susceptibility testing on clinical isolates. In our study, we identified one strain of A. pleuropneumoniae with resistance to florfenicol and extracted a 5919 bp plasmid named pAPPJY, which confers florfenicol resistance. Sequence analysis revealed that the plasmid contains four open reading frames, namely rep, antioxin vbha family protein, floR, and a partial copy of lysr. Although a few variations in gene position were observed, the plasmid sequence exhibits a high degree of similarity to other florfenicol-resistant plasmids found in Glaesserella parasuis and A. pleuropneumoniae. Therefore, it is possible that the pAPPJY plasmid functions as a shuttle, facilitating the spread of florfenicol resistance between G. parasuis and A. pleuropneumoniae. In addition, partial recombination may occur during bacterial propagation. In conclusion, this study highlights the horizontal transmission of antibiotic resistance among different bacterial species through plasmids, underscoring the need for increased attention to antibiotic usage. [ABSTRACT FROM AUTHOR]

Introduction: Carp edema virus (CEV) is a fish poxvirus that primarily infects the gills of common carp. CEV causes koi sleepy disease (KSD), which is highly contagious and can result in mortality of up to 100%. Methods: In the present study, we analyzed the stress and immune responses during KSD in two strains of common carp with different resistance to CEV: susceptible koi and resistant Amur sazan. Experiments were performed at two temperatures: 12°C and 18°C. In the case of koi carp, we also analyzed the effect of supplementation of 0.6% NaCl into tank water, which prevents mortality of the CEV-infected fish (salt rescue model). Results: We found that CEV-infected koi kept at 18°C had the highest viral load, which correlated with the most severe histopathological changes in the gills. CEV infection resulted in the activation of stress response reflected by the upregulated expression of genes involved in stress response in the stress axis organs and increased levels of cortisol and glucose in the blood plasma. These changes were the most pronounced in CEV-infected koi kept at 18°C. At both temperatures, the activation of antiviral immune response was observed in koi kept under freshwater and NaCl conditions upon CEV infection. Interestingly, a clear downregulation of the expression of adaptive immune genes was observed in CEV-infected koi kept under freshwater at 18°C. Conclusion: CEV induces a stress response and modulates adaptive immune response in koi, and this is correlated with the level of viral load and disease development. [ABSTRACT FROM AUTHOR]

Porcine reproductive and respiratory syndrome (PRRS) is one of the most economically important infectious diseases for the pig industry worldwide. The disease was firstly reported in 1987 and became endemic in many countries. Since then, outbreaks caused by strains of high virulence have been reported several times in Asia, America and Europe. Interstitial pneumonia, microscopically characterised by thickened alveolar septa, is the hallmark lesion of PRRS. However, suppurative bronchopneumonia and proliferative and necrotising pneumonia are also observed, particularly when a virulent strain is involved. This raises the question of whether the infection by certain strains results in an overstimulation of the proinflammatory response and whether there is some degree of correlation between the strain involved and a particular pattern of lung injury. Thus, it is of interest to know how the inflammatory response is modulated in these cases due to the interplay between virus and host factors. This review provides an overview of the macroscopic, microscopic, and molecular pathology of PRRSV-1 strains in the lung, emphasising the differences between strains of different virulence. [ABSTRACT FROM AUTHOR]

Gentamicin, an aminoglycoside antibiotic, is a mixture of therapeutically active C1, C1a, C2 and other minor components. Despite its decades-long use in pigs and other species, its intramuscular (IM) pharmacokinetics/pharmacodynamics (PKs/PDs) are unknown in piglets. Furthermore, the PKs of many drugs differ between healthy and sick animals. Therefore, we investigated the PKs of gentamicin after a single IM dose (10 mg/kg) in healthy piglets and piglets that were intranasally co-infected with Actinobacillus pleuropneumoniae and Pasteurella multocida (PM). The plasma concentrations were measured using validated liquid chromatography/mass spectrometry. The gentamicin exposure was 36% lower based on the area under the plasma concentration–time curve and 16% lower based on the maximum plasma concentration (Cmax) in the infected piglets compared to the healthy piglets, while it was eliminated faster (shorter half-life and larger clearance) in the infected piglets compared to the healthy piglets. The clearance and volume of distribution were the highest for the C1 component. C1, C1a and C2 accounted for 22–25%, 33–37% and 40–42% of the total gentamicin exposure, respectively. The PK/PD target for the efficacy of aminoglycosides (Cmax/minimum inhibitory concentration (MIC) > 10) could be exceeded for PM, with a greater magnitude in the healthy piglets. We suggest integrating this PK information with antibiotic susceptibility data for other bacteria to make informed antibiotic and dosage regimen selections against piglet infections. [ABSTRACT FROM AUTHOR]

Background: Porcine circovirus type 2 (PCV2) is a globally prevalent and recurrent pathogen that primarily causes slow growth and immunosuppression in pigs. Porcine circovirus type 3 (PCV3), a recently discovered virus, commonly leads to reproductive disorders in pigs and has been extensively disseminated worldwide. Infectionwith a single PCV subtype alone does not induce severe porcine circovirusassociated diseases (PCVD), whereas concurrent co-infection with PCV2 and PCV3 exacerbates the clinical manifestations. Pseudorabies (PR), a highly contagious disease in pigs, pose a significant threat to the swine industry in China. Methods: In this study, recombinant strains named rPRV-2Cap/3Cap and rPRV-2Cap/3Cap/IL4 was constructed by using a variant strain XJ of pseudorabies virus (PRV) as the parental strain, with the TK/gE/gI genes deleted and simultaneous expression of PCV2 Cap, PCV3 Cap, and IL-4. The two recombinant strains obtained by CRISPR/Cas gE gene editing technology and homologous recombination technology has genetic stability in baby hamster Syrian kidney-21 (BHK-21) cells and is safe to mice. Results: rPRV-2Cap/3Cap and rPRV-2Cap/3Cap/IL4 exhibited good safety and immunogenicity in mice, inducing high levels of antibodies, demonstrated 100% protection against the PRV challenge in mice, reduced viral loads and mitigated pathological changes in the heart, lungs, spleen, and lymph nodes during PCV2 challenge. Moreover, the recombinant viruses with the addition of IL-4 as a molecular adjuvant outperformed the non-addition group in most indicators. Conclusion: rPRV-2Cap/3Cap and rPRV-2Cap/3Cap/IL4 hold promise as recombinant vaccines for the simultaneous prevention of PCV2, PCV3, and PRV, while IL-4, as a vaccine molecular adjuvant, effectively enhances the immune response of the vaccine. [ABSTRACT FROM AUTHOR]

Background: The objective of this study was to examine the inter-relationships between pig farm management and facilities (as assessed by questionnaire) and post-mortem lung lesion (lung score assesment), which are the result of respiratory infections. The relationships between carcass characteristics and post-mortem lung lesion scores were also investigated. Results: Questionnaire responses were collected from 22 self-selecting pig farmers about their farm facilities/management and health condition of the respiratory system of pigs, including the occurrence of clinical respiratory signs, results of laboratory testing for respiratory pathogens, and the use of respiratory vaccines. When fatteners were sent to the abattoir, their carcasses (n = 1,976) were examined for evidence of respiratory disease by lung lesion (pleuritis pneumonia-like (PP-like) and enzootic pneumonia-like (EP-like) lesions) scoring and the Actinobacillus pleuropneumoniae Index (APPI) was calculated. Carcass characteristics were recorded and, retrospectively, the prevalence of cachectic pigs was calculated. Using these variables, the relationships between farm facilities/management and lung lesions scores and the relationships between the latter and carcass characteristics and cachexia were explored. The key findings relating farm facilities and management to lung lesions were: slatted floors were associated with significantly higher EP-like lesions scores than litter bedding in weaners, single-stage fattening in the same building was associated with significantly higher EP-like lesions scores than two-stage fattening, but herd size, stocking density, use of all-in/all-out (AIAO) rule, technological break duration and variation in daily temperature did not affect lung lesions scores. The key findings relating lung lesion scores to carcass characteristics were: a significant, negative correlation between EP-like scores and carcass weight but not with other carcass characteristics, a significant positive correlation between PP-like scores and carcass meat content and prevalence of cachectic carcasses and a significant positive correlation between lung APPI and prevalence of cachectic carcasses. Conclusions: It can be concluded that both farm facilities and management affect lung lesions scores and that the latter affect carcass characteristics. Lung lesion scoring is an inexpensive technique suitable for rapid monitoring of large numbers of carcasses that can be performed after animal slaughter. It provides useful information to inform producers about possible deficits in farm facilities or management and is a predictor of economic loss due to poorer quality carcasses. [ABSTRACT FROM AUTHOR]

African swine fever virus represents a significant reemerging threat to livestock populations, as its incidence and geographic distribution have surged over the past decade in Europe, Asia, and Caribbean, resulting in substantial socio-economic burdens and adverse effects on animal health and welfare. In a previous report, we described the protective properties of our newly thermo-attenuated strain (ASFV- 989) in pigs against an experimental infection of its parental Georgia 2007/1 virulent strain. In this new study, our objective was to characterize the molecular mechanisms underlying the attenuation of ASFV-989. We first compared the activation of type I interferon pathway in response to ASFV-989 and Georgia 2007/1 infections, employing both in vivo and in vitro models. Expression of IFN-a was significantly increased in porcine alveolar macrophages infected with ASFV-989 while pigs infectedwith Georgia 2007/1 showed higher IFN-a than those infected by ASFV-989. We also used a medium-throughput transcriptomic approach to study the expression of viral genes by both strains, and identified several patterns of gene expression. Subsequently, we investigated whether proteins encoded by the eight genes deleted in ASFV-989 contribute to the modulation of the type I interferon signaling pathway. Using different strategies, we showed that MGF505-4R interfered with the induction of IFN-α/β pathway, likely through interaction with TRAF3. Altogether, our data reveal key differences between ASFV-989 and Georgia 2007/ 1 in their ability to control IFN-α/β signaling and provide molecular mechanisms underlying the role of MGF505-4R as a virulence factor. [ABSTRACT FROM AUTHOR]

Introduction: Influenza A viruses (IAVs) infect the respiratory tract of mainly humans, poultry, and pigs. Co-infections with pathogenic lung bacteria are a common event and contribute to the severity of disease progression. Neutrophils are a major cell type of the innate immune system and are rapidly recruited to the site of infection. They have several effector functions to fight invading pathogens such as the secretion of reactive oxygen species (ROS) or the release of neutrophil extracellular traps (NETs). NETs are known to promote the growth of Pasteurellaceae bacteria, especially if degraded by nucleases. Methods: In this study, bronchoalveolar lavage fluid (BALF) from 45 field-infected pigs was analyzed for 1) NET markers, 2) influence on growth of lung bacteria, and 3) impact on neutrophil functions. BALF samples from 21 IAV-positive pigs and 24 lung diseased but IAV-negative pigs were compared. Results: Here, we show that neutrophils in the lungs of IAV-positive pigs release vesicular NETs. Several NET markers were increased in the BALF of IAV-positive pigs compared with the BALF from IAV-negative pigs. The amount of NET markers positively correlated with the viral load of the IAV infection. Interestingly, the BALF of IAV-positive pigs enhanced the growth of bacteria belonging to the family of Pasteurellaceae as potential coinfecting bacteria. These effects were weaker with the BALF derived from IAV-negative pigs with other lung infections. The intensity of oxidative burst in neutrophils was significantly decreased by BALF from IAVpositive pigs, indicating impaired antimicrobial activity of neutrophils. Finally, the lung milieu reflected by IAVpositive BALF does not enable neutrophils to kill Actinobacillus pleuropneumoniae but rather enhances its growth. Discussion: In summary, our data show that an IAV infection is affecting neutrophil functions, in particular the release of NETs and ROS. Furthermore, IAV infection seems to provide growth-enhancing factors for especially coinfecting Pasteurellaceae and reduces the killing efficiency of neutrophils. [ABSTRACT FROM AUTHOR]

Actinobacillus pleuropneumoniae is the causative agent of pleuropneumonia, an economically important lung disease in pigs. In draft genomes of two Cypriot clinical A. pleuropneumoniae isolates (MIDG3457 and MIDG3459), we previously identified single genomic regions with homology to Mu-like bacteriophage and presented preliminary evidence of active phage. Here, updated Phastest genomic analysis identified two loci in both MIDG3457 and MIDG3459 that were predicted to encode proteins with high homology to, and whose organisation was characteristic of, Mu-like phages. Phylogenetically, the closest matches were with Mannheimia Vb and Glaesserella SuMu phages. Phastest scored the loci as "complete", indicating they produced active phage. PCR amplification of the Mu-like phage c and tail genes from DNase-treated polyethylene glycol 8000 (PEG)-precipitated supernatants of MIDG3457 and MIDG3459 (grown in either Brain Heart Infusion-NAD or Grace's Insect Medium-NAD broth) indicated the presence of intact virions. The phages from MIDG3457 and MIDG3459 were named PluMu 3457-1, 3457-2, and PluMu 3459-1 and PluMu 3459-2, respectively. Transmission electron microscopy (TEM) of the PEG-precipitated supernatants of broth-grown MIDG3459 identified virions with icosahedral heads and tails, consistent with other Mu-like phages. We conclude that MIDG3459 produces an active Mu-like phage. [ABSTRACT FROM AUTHOR]

Objective: The objective of this study was to characterize ICEAplChn2, a novel SXT/R391-related integration and conjugation element (ICE) carrying 19 drug resistance genes, in a clinical isolate of Actinobacillus pleuropneumoniae from swine. Methods: Whole genome sequencing (WGS) of A. pleuropneumoniae CP063424 strain was completed using a combination of third-generation PacBio and second-generation Illumina. The putative ICE was predicted by the online tool ICEfinder. ICEAplChn2 was analyzed by PCR, conjugation experiments, and bioinformatics tools. Results:A. pleuropneumoniae CP063424 strain exhibited high minimum inhibitory concentrations of clindamycin (1,024 mg/L). The WGS data revealed that ICEAplChn2, with a length of 167,870 bp and encoding 151 genes, including multiple antibiotic resistance genes such as erm(42), VanE, LpxC, dfrA1, golS, aadA3, EreA, dfrA32, tetR(C), tet(C), sul2, aph(3)″-lb, aph(6)-l, floR, dfrA, ANT(3″)-IIa, catB11, and VanRE, was found to be related to the SXT/R391 family on the chromosome of A. pleuronipneumoniae CP063424. The circular intermediate of ICEAplChn2 was detected by PCR, but conjugation experiments showed that it was not self-transmissible. Conclusions: To our knowledge, ICEAplChn2 is the longest member with the most resistance genes in the SXT/R391 family. Meanwhile, ATP-binding cassette superfamily was found to be inserted in the ICEAplChn2 and possessed a new insertion region, which is the first description in the SXT/R391 family. [ABSTRACT FROM AUTHOR]

Pradofloxacin is a dual targeting, bactericidal fluoroquinolone recently approved for treating bacteria causing swine respiratory disease. Currently, an abundance of in vitro data does not exist for pradofloxacin. We determined the minimum inhibitory concentration (MIC) and mutant prevention concentrations (MPC) of pradofloxacin compared to ceftiofur, enrofloxacin, florfenicol, marbofloxacin, tildipirosin, tilmicosin and tulathromycin against swine isolates of Actinobacillus pleuropneumoniae and Pasteurella multocida. Overall, pradofloxacin had the lowest MIC and MPC values as compared to the other agents tested. For example, pradofloxacin MIC values for 50%, 90% and 100% of A. pleuropneumoniae strains were ≤0.016 µg/mL, ≤0.016 µg/mL and ≤0.016 µg/mL and for P. multocida were ≤0.016 µg/mL, ≤0.016 µg/mL and 0.031 µg/mL, respectively. The MPC values for 50%, 90% and 100% of A. pleuropneumoniae strains were 0.031 µg/mL, 0.063 µg/mL and 0.125 µg/mL and for P. multocida were ≤0.016 µg/mL, 0.031 µg/mL and 0.0.063 µg/mL, respectively. By MPC testing, all strains were at or below the susceptibility breakpoint. Based on MPC testing, pradofloxacin appears to have a low likelihood for resistance selection. This study represents the most comprehensive in vitro comparison of the above noted drugs and the first report for pradofloxacin and tildipirosin.

A large variety of clinical manifestation in individual pigs occurs after infection with pathogens involved in porcine respiratory disease complex (PRDC). Some pigs are less prone to develop respiratory disease symptoms. The variation in clinical impact after infection and the recovery capacity of an individual animal are measures of its resilience. In this paper, we examined which ones of a range of animal-based factors (rectal temperature, body weight, skin lesion scores, behavior, natural antibody serum levels, serum levels of white blood cells, and type of T and granulocyte subsets) when measured prior to infection are related to disease severity. These animalbased factors and the interaction with housing regimen of the piglets (conventional or enriched) were modeled using linear regression to predict disease severity using a dataset acquired from a previous study using a well-established experimental coinfection model of porcine reproductive and respiratory syndrome virus (PRRSV) and Actinobacillus pleuropneumoniae. Both PRRSV and A. pleuropneumoniae are often involved in PRDC. Histological lung lesion score of each animal was used as a measure for PRDC severity after infection. Prior to infection, higher serum levels of lymphocytes (CD3+), naïve T helper (CD3+CD4+CD8−), CD8+ (as well as higher relative levels of CD8+), and memory T helper (CD3+CD4+CD8+) cells and higher relative levels of granulocytes (CD172a ) were related to reduced disease severity in both housing systems. Raised serum concentrations of natural IgM antibodies binding to keyhole limpet hemocyanin (KLH) were also related to reduced disease severity after infection. Increased levels of skin lesions at the central body part (after weaning and before infection) were related to increased disease severity in conventional housing systems only. High resisters showed a lower histological lung lesion score, which appeared unrelated to sex. Body temperature, behavior, and growth prior to infections were influenced by housing regimen but could not explain the variation in lung lesion scores after infection. Raised basal lymphocyte counts and lower skin lesion scores are related to reduced disease severity independent of or dependent on housing system, respectively. In conclusion, our study identifies intrinsic animal-based measures using linear regression analysis that predicts resilience to infections in pigs. [ABSTRACT FROM AUTHOR]

Simple Summary: The Spanish swine sector has experienced continuous growth during recent years, becoming the largest pig sector in Europe and one of the largest exporters of pig meat worldwide. One of the main challenges for the stability and productivity of the Spanish swine industry is the presence of several swine diseases, resulting in an important economic impact. Information on the current frequency and importance of the main swine diseases which affect the Spanish pig industry in each phase could help in optimizing and prioritizing the efforts within disease control programs. This study described the frequency and importance of different pathogens for veterinarians and consultants in the production phases, as well as the most used tools for controlling such pathogens and diseases. One of the main challenges for the sustainability and productivity of the Spanish swine industry is health instability, resulting in significant economic losses. Information on the main swine diseases which affect the Spanish pig industry could help in optimizing the efforts within control programs. This study determined the frequency of occurrence of the main diseases in Spain and the main control tool used, based on perceptions from veterinarians and consultants in a specific survey. Results showed that Streptococcus (S.) suis, E. coli, and coccidia are the most frequent pathogens in the gestation and lactation phase, whereas the most important were Porcine Reproductive and Respiratory Syndrome virus (PRRSV). In the nursery phase, the most frequent were S. suis, E. coli, and PRRSV, the latter being the most important for the participants. Finally, in the fattening phase, PRRSV and Actinobacillus pleuropneumoniae were the most frequent and important pathogen, respectively. Statistical differences among responses were detected with respect to the location and the gestation and lactation phases by farm size. Regarding the tools used for controlling the diseases, vaccination was the main strategy in all production phases, except in the fattening period, in which antibiotherapy was the most common response from the participants. Finally, the improvement of management practices was the most proposed tool, suggesting its importance within control programs. [ABSTRACT FROM AUTHOR]

Simple Summary: Actinobacillus (A.) pleuropneumoniae is a major agent of the porcine respiratory diseases complex (PRDC), together with several bacterium and virus species. A. pleuropneumoniae strains are diverse regarding virulence and antigen structure; they have different virulence variants, and the strains can be assigned into 19 serovars. Serovar 16 was described in Hungary for the first time, and it is relatively frequent in pig herds. The aim of the present work was examination of the virulence of the A. pleuropneumoniae A-85/14 strain, the type strain of serovar 16, by infecting piglets. The results prove that the A. pleuropneumoniae A-85/14 strain is a virulent one. Different virulence variants of A. pleuropneumoniae are involved in the etiology of porcine pleuropneumonia. The purpose of the present trial was examination of the virulence of the Actinobacillus pleuropneumoniae A-85/14 strain, the type strain of serovar 16, in an animal challenge experiment. Thirty 12-week-old piglets seronegative for A. pleuropneumoniae were allocated into three trial groups each of 10 animals, and they were infected intranasally with 106, 107, or 108 colony forming units (cfu) of the strain, respectively. Clinical signs were recorded twice a day, and the animals were euthanized 6 days after the infection. Typical clinical signs and postmortem lesions of porcine pleuropneumonia were seen in the animals of each trial group; however, they were generally mild, and no significant differences could be seen between the three groups. Even 106 colony forming units of A. pleuropneumoniae A-85/14 strain could induce clinical signs and lesions. Based on these results, the type strain of serovar 16 of A. pleuropneumoniae must be regarded as a typical pathogenic strain of the species. [ABSTRACT FROM AUTHOR]

Introduction: Feline parvovirus (FPV), a single-stranded DNA virus, is accountable for causing feline panleukopenia, a highly contagious and often lethal disease that primarily affects cats. The epidemiology prevalence and pathogenicity of FPV in certain regions of China, however, remains unclear. The aim of this research was to investigate the epidemiology of FPV in different regions of China in 2021 and compare its infectivity and pathogenicity. Methods: In this research, a total of 36 FPV strains were obtained from diverse regions across China. Phylogenetic analysis was performed based on the VP2 and NS1 sequences, and two representative strains, FPV027 and FPV072, which belonged to different branches, were selected for comparative assessment of infectivity and pathogenicity. Results and discussion: The results revealed that all strains were phylogenetically classified into two groups, G1 and G2, with a higher prevalence of G1 strains in China. Both in vitro and in vivo experiments demonstrated that FPV072 (G1 group) exhibited enhanced infectivity and pathogenicity compared to FPV027 (G2 Group). The structural alignment of the VP2 protein between the two viruses revealed mutations in residues 91, 232, and 300 that may contribute to differences in infectivity and pathogenicity. The findings from these observations will contribute significantly to the overall understanding of the molecular epidemiology of FPV in China and facilitate the development of an effective FPV vaccine. [ABSTRACT FROM AUTHOR]