• Journal of Microbiology and Biotechnology
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• v.22 no.9
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• pp.1307-1309
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• 2012

Haemophilus parasuis causes contagious porcine Gl$\ddot{a}$sser's disease leading to severe losses in the swine industry. In this study, we established an efficient Escherichia coli-based system for the expression of H. parasuis major outer-membrane protein (MOMP) that has been known as a good vaccine candidate against Gl$\ddot{a}$sser's disease. Use of an E. coli-derived pelB leader sequence made it possible to produce recombinant MOMP (rMOMP) as the soluble forms without an additional refolding process. Using two different animal models, it was evaluated that the rMOMP was capable of inducing a significant immune response and providing protection against H. parasuis infection.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.843-852
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• 1998

From Jan. 1996 to Oct. 1997, 29 pigs with 40-70 days old showing dyspnea inappetite and polyserositis were collected and carried out necropsy, bacterial culture, histopathology, avidin biotin complex(ABC) stain, fluorescent antibody(FA) test, and electron microscopy. In the study, 4 strains from 3 pigs were isolated from meninges, pleura and synovial fluid and also were identified as Haemophilus parasuis serovar 5. Main histopathological lesions of 29 pigs with polyserositis were frequently composed of fibrinous peritonitis(27), pleurisy(22), interstitial pneumonia(21), fibrinous epicarditis(20), fibrinopurulent meningitis(8) and synovitis(4). By ABC stain, 11/29(37.9%) pigs with polyserositis were confirmed to have H parasuis serovar 5 in the cytoplasm of macrophages and neutrophils in cerebral meninges, epicardium, pleura surface of lung or serosa of spleen. ABC stain(20.8~40.0%) to detect H parasuis serovar 5 in tissues was more sensitive than bacterial culture(10.3%), but less sensitive than FA test(62.5%) using frozen tissues even though the result of 8 cases. By electron microscopy, a bacterium was also detected in the cytoplasm of macrophages in purulent exudate of cerebral meninges. Consequently, we confirmed that H parasuis serovar 5 has been involving to cause pigs with polyserositis and can be detected by FA and ABC stain as reliable diagnostic tools.

• Journal of Veterinary Science
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• v.22 no.5
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• pp.59.1-59.13
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• 2021

Background: Antimicrobial peptides (AMPs) have been identified as promising compounds for consideration as novel antimicrobial agents. Objectives: This study analyzed the efficacy of cecropin B against Haemophilus parasuis isolates through scanning electron microscopy (SEM) and atomic force microscopy (AFM) experiments. Results: Cecropin B exhibited broad inhibition activity against 15 standard Haemophilus parasuis (HPS) strains and 5 of the clinical isolates had minimum inhibition concentrations (MICs) ranging from 2 to 16 ㎍/mL. Microelectrophoresis and hexadecane adsorption assays indicated that the more hydrophobic and the higher the isoelectric point (IEP) of the strain, the more sensitive it was to cecropin B. Through SEM, multiple blisters of various shapes and dents on the cell surface were observed. Protrusions and leakage were detected by AFM. Conclusions: Based on the results, cecropin B could inhibit HPS via a pore-forming mechanism by interacting with the cytoplasmic membrane of bacteria. Moreover, as cecropin B concentration increased, the bacteria membrane was more seriously damaged. Thus, cecropin B could be developed as an effective anti-HPS agent for use in clinical applications.

• Journal of Veterinary Science
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• v.23 no.1
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• pp.2.1-2.18
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• 2022

Background: Co-infections of the porcine reproductive and respiratory syndrome virus (PRRSV) and the Haemophilus parasuis (HPS) are severe in Chinese pigs, but the immune response genes against co-infected with 2 pathogens in the lungs have not been reported. Objectives: To understand the effect of PRRSV and/or HPS infection on the genes expression associated with lung immune function. Methods: The expression of the immune-related genes was analyzed using RNA-sequencing and bioinformatics. Differentially expressed genes (DEGs) were detected and identified by quantitative real-time polymerase chain reaction (qRT-PCR), immunohistochemistry (IHC) and western blotting assays. Results: All experimental pigs showed clinical symptoms and lung lesions. RNA-seq analysis showed that 922 DEGs in co-challenged pigs were more than in the HPS group (709 DEGs) and the PRRSV group (676 DEGs). Eleven DEGs validated by qRT-PCR were consistent with the RNA sequencing results. Eleven common Kyoto Encyclopedia of Genes and Genomes pathways related to infection and immune were found in single-infected and co-challenged pigs, including autophagy, cytokine-cytokine receptor interaction, and antigen processing and presentation, involving different DEGs. A model of immune response to infection with PRRSV and HPS was predicted among the DEGs in the co-challenged pigs. Dual oxidase 1 (DUOX1) and interleukin-21 (IL21) were detected by IHC and western blot and showed significant differences between the co-challenged pigs and the controls. Conclusions: These findings elucidated the transcriptome changes in the lungs after PRRSV and/or HPS infections, providing ideas for further study to inhibit ROS production and promote pulmonary fibrosis caused by co-challenging with PRRSV and HPS.

• Korean Journal of Veterinary Service
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• v.31 no.3
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• pp.347-356
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• 2008

The purpose of this study was to evaluate the efficacy and cross-protection of serovar 12 against serovar 4 or 5 of H parasuis with M+$Parapac^{(R)}$. A total of 141 piglets from 2(A and B) farms were used and divided into experimental group and control group in each farm. Farm A has been detected H parasuis serovar 12, whereas farm B has been detected H parasuis serovar 4 or 5 from post-weaned pigs with PMWS. The piglets were vaccinated intramuscularly with 2.0ml of M+$Parapac^{(R)}$ in experimental group or normal saline in control group at 1 week of age. A same booster dose was given at 3 weeks of age. In order to value the antibody titer to H parasuis using by tube agglutination test, blood samples were collected from piglets at the aged of 1 week, 6 and 14 weeks. In experimental group and control group, the average antibody titers were $32.5{\pm}21.0,\;114.5{\pm}34.0,\;98.1{\pm}55.4$ and $32.9{\pm}18.6,\;25.8{\pm}36.9,\; 746.7{\pm}1,215.8$ at the aged of 1 week, 6 and 14 weeks, respectively. The cumulative clinical sign indexes by standard guideline of Schering-Plough Animal Health Corp were 486 and 1,069, respectively. The average daily gains and feed conversion rates were $0.553{\pm}0.016kg$ and $0.492{\pm}0.004kg$, and 1.99 and 2.24, respectively. The average gross lesion scores were $1.0{\pm}0.8$ and $1.9{\pm}0.6$, respectively. According to these results, the M+$Parapac^{(R)}$ containing H parasuis serovar 12 may be induce circulating antibodies that cross-react with serovar 4 or 5 and have a protection of PMWS with H parasuis.

• Journal of Veterinary Clinics
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• v.11 no.2
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• pp.507-516
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• 1994

The purposes of this investigations were survey of the occurred bacterial diseases, development of new animal health drug, guidance to formers on the treatment and control methods of diseases. Some series of investigations have been carried out by microbiological, pathological and serological examinations. The results could be summarized as follows. 1. A total of 953 cases of outbreaked swine diseases have been diagnosed in Clinical pathology laboratories, Bayer Vet Res Institute during 8 years (from 1986 to 1993). The high incidence diseases were colibacillosis, pleuropneumonia, streptococcal infection and pasteurellosis in decreasing order. 2. Pleuropneumonia caused by Actinobacillus pleuropneumoniae was the most important respiratory diseases and pasteurellosis by Pasteurella multocide could be confirmed in several cases. 3. Actinobacillus pleuropneumoniae 50 strains were isolated and identified by biochemical and serological tests. In serotyping test, 22 isolated strains were serotype 5, 21 strains as serotype 2, each 2 strains as serotype 3 and 7 by the coagglutination test. 4. Colibacillosis and edema discase caused by Escherichia coli has been the most predominant outbreaked disease in this investigations. The 100 isolates of E coli strains were sensitive to amikacin, colistin, enrofloxacin, gentamycin and trimethoprim -sulfamethoxazole. 5. Swine erysipelas caused by Erysipelothrix rhusiopathiae was confirmed 25 cases as acute septicemic forms. Isolates of E rhusiopathiae were highly sensitive to ampicillin, cephalothin, enrofloxcin, penicillin and tetracycline. 6. The 49 cases of hemorrhagic and necrotic enteritis in piglets were observed and 13 strains of Clostridium perfringens could be isolated and confirmed by biological and serological test. Isolates of Clostridium perfringens type C were highly sensitive to ampicillin, cephalothin, enrofloxacin, penicillin and trimethoprim- sulfamethoxazole. 7. The 14 strains of Streptococcus suis type II could be isolated from meningitis of piplets. 8. Polyserositis caused by Haemophilus parasuis and salmonellosis were observed and confirmed. Also Corynebacterial infections and several parasitosis have been also observed in this investigations.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.4
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• pp.576-584
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• 2017

Objective: To generate recombinant Bacillus subtilis (B. subtilis) engineered for expression of porcine ${\beta}-defensin-2$ (pBD-2) and cecropin P1 (CP1) fusion antimicrobial peptide and investigate their anti-bacterial activity in vitro and their growth-promoting and disease resisting activity in vivo. Methods: The pBD-2 and CP1 fused gene was synthesized using the main codons of B. subtilis and inserted into plasmid pMK4 vector to construct their expression vector. The fusion peptide-expressing B. subtilis was constructed by transformation with the vector. The expressed fusion peptide was detected with Western blot. The antimicrobial activity of the expressed fusion peptide and the recovered pBD-2 and CP1 by enterokinase digestion in vitro was analyzed by the bacterial growth-inhibitory activity assay. To analyze the engineered B. subtilis on growth promotion and disease resistance, the weaned piglets were fed with basic diet supplemented with the recombinant B. subtilis. Then the piglets were challenged by enteropathogenic Escherichia coli (E. coli). The weight gain and diarrhea incidence of piglets were measured after challenge. Results: The recombinant B. subtilis engineered for expression of pBD-2/CP1 fusion peptide was successfully constructed using the main codons of the B. subtilis. Both expressed pBD-2/CP1 fusion peptide and their individual peptides recovered from parental fusion peptide by enterokinase digestion possessed the antimicrobial activities to a variety of the bacteria, including gram-negative bacteria (E. coli, Salmonella typhimurium, and Haemophilus parasuis) and grampositive bacteria (Staphylococcus aureus). Supplementing the engineered B. subtilis to the pig feed could significantly promote the piglet growth and reduced diarrhea incidence of the piglets. Conclusion: The generated B. subtilis strain can efficiently express pBD-2/CP1 fusion antimicrobial peptide, the recovered pBD-2 and CP1 peptides possess potent antimicrobial activities to a variety of bacterial species in vitro. Supplementation of the engineered B. subtilis in pig feed obviously promote piglet growth and resistance to the colibacillosis.

• Korean Journal of Veterinary Service
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• v.34 no.4
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• pp.313-320
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• 2011

The purpose of this study was to investigate association with gross lesions and causative pathogens of porcine respiratory disease complex (PRDC) including porcine circovirus type 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza virus (SIV), Mycoplasma hyopneumoniae (MH), Pasteurella multocida (PM), Actinobacillus pleuropneumoniae (APP), Haemophilus parasuis (HP) in slaughtered pigs. A total of 1,200 lung samples were collected randomly from slaughtered pigs in Korea during August of 2010 through July of 2011. The gross lesions were classified according to the six stages (0, 1~10, 11~20, 21~30, 31~40 and ${\geq}41$, unit=%) and 48 samples from each stage were selected to detect viral and bacterial pathogens. The results according to the six stages were 100 (8.3%), 259 (21.6%), 326 (27.2%), 213 (17.8%), 144 (12.0%) and 158 (13.2%) cases, respectively. Prevalence of pneumonia according to season was 87.0~96.7% and the highest prevalence was in spring. In detection of pathogens by PCR, 53 samples were not detected any causative pathogens of PRDC. PCV2, PRRSV, SIV, MH, PM, APP serotype 2, APP serotype 5 and HP were positive in 45.5%, 12.5%, 10.4%, 60.1%, 1.7%, 13.9%, 12.2% and 15.6%, respectively. In co-infection, PCV2-MH was the most detected causative pathogens of PRDC. The detection rate of PCV2 and PRRSV was the highest in spring, of SIV, MH and HP was in winter. The detection rate of APP-2 and APP-5 had no seasonal prevalence. The more severe gross lesions increased, the higher the detection rate showed.