Mycoplasma Hyopneumoniae Vaccine Research Articles

Drug Susceptibility Test and Analysis of Quinolone Resistance Genes in Mycoplasma hyopneumoniae Vaccine Strains and Field Isolates from China.

Enrofloxacin is a commonly used animal-specific drug in veterinary clinics. However, this drug has no epidemiological cutoff values (ECVs/ECOFFs) for Mycoplasma hyopneumoniae in CLSI and EUCAST. Defining the epidemiological cutoff values (ECOFFs) of enrofloxacine to Mycoplasma hyopneumoniae (M. hyo) can inform an early detection of bacterial resistance to better manage the resistance prevention and also help in establishing drug resistance breakpoints;. We determined the susceptibility breakpoint of M. hyo to enrofloxacin by the American Clinical and Laboratory Standards Institute (CLSI) standard method based on the PCR of vaccine strains and wild strains drug resistance genes;. Eighty strains of M.hyo isolated in Tibet were moderately sensitive (S) to tetracycline, florfenicol, spiramycin, erythromycin thiocyanate, tilmicosin, tiamulin, lincomycin, clindamycin, ofloxacin, enrofloxacin, gentamicin, amikacin, with MICs below 0.5 μg/mL. For vaccine 168L, RM48, and J strains, the susceptibility to the same antibacterial drugs was lower compared to the Tibetan isolates. The resistance of J strain to erythromycin thiocyanate was confirmed. Gene point mutation was confirmed in Quinolone Resistance Determining Regions (QRDR) of HNSH strain Topoisomerase IV subunit A, this finding is compared with the sequencing results of 168L strain reference sequence (Accession number: CP003131). Arg-Lys amino acid mutation (G921A and G1179A) was confirmed for the increase of MIC value involved in M.hyo to enrofloxacine;. The cut-off value of M.hyo to enrofloxacin was set as 1 μg/mLby ECOFFinder XL 2010 V2.1.

Phenotypic characteristics and protective efficacy of an attenuated Mycoplasma hyopneumoniae vaccine by aerosol administration

With the improvement of large-scale breeding in pig farms, conventional head-by-head immunization has disadvantages with low efficiency and high cost. Considering that most pathogens leading to pulmonary diseases circulate from the respiratory mucosa, immunization through the respiratory tract route has been a highly attractive vaccine delivery strategy. In this study, to develop an effective Mycoplasma hyopneumoniae (Mhp) aerosol vaccine, a customized ultrasonic atomizer was developed. The aerodynamic diameter, activity, and content of the Mhp aerosol vaccine were measured. In addition, piglets were immunized with the Mhp aerosol vaccine, and the immunity of the animal challenge protection test was evaluated. At the end of nebulization, the mass median aerodynamic diameters (MMAD) and geometric standard deviation (GSD) of the aerosol were 2.98 ± 0.02 μm and 1.51 ± 0.02, respectively. Moreover, 10 min after nebulization, the MMAD and GSD of the aerosol were 2.76 ± 0.02 μm and 1.51 ± 0.01, respectively, which were hardly changed. Compared with theoretical value, the actual titer of aerosol vaccines presented in 50% color changing unit (CCU50) after nebulization decreased 0.6. The shape, size, and uniformity of collected aerosols are relatively stable. The proportion of Mhp in aerosol produced by vaccine stock solution and 10 times diluted vaccine solution was 76.52% and 58.82%, respectively, and the average number of Mhp in a single aerosol was 3.06 and 1.51, respectively. In addition, the aerosol vaccine antigen particles could be transported to the lower respiratory tract, a local mucosal immune response was induced in piglets. The vaccine colonized the respiratory tract and significantly decline the lung lesion index after aerosol vaccination. In conclusion, an effective aerosol vaccine against Mhp infection was developed. And this is the first effective assessment for Mhp live vaccine with aerosolization against infection in piglets.

Signaling differences in peripheral blood mononuclear cells of high and low vaccine responders prior to, and following, vaccination in piglets

Individual variability in responses to vaccination can result in vaccinated subjects failing to develop a protective immune response. Vaccine non-responders can remain susceptible to infection and may compromise efforts to achieve herd immunity. Biomarkers of vaccine unresponsiveness could aid vaccine research and development as well as strategically improve vaccine administration programs. We previously vaccinated piglets (n = 117) against a commercial Mycoplasma hyopneumoniae vaccine (RespiSure-One) and observed in low vaccine responder piglets, as defined by serum IgG antibody titers, differential phosphorylation of peptides involved in pro-inflammatory cytokine signaling within peripheral blood mononuclear cells (PBMCs) prior to vaccination, elevated plasma interferon-gamma concentrations, and lower birth weight compared to high vaccine responder piglets. In the current study, we use kinome analysis to investigate signaling events within PBMCs collected from the same high and low vaccine responders at 2 and 6 days post-vaccination. Furthermore, we evaluate the use of inflammatory plasma cytokines, birthweight, and signaling events as biomarkers of vaccine unresponsiveness in a validation cohort of high and low vaccine responders. Differential phosphorylation events (FDR < 0.05) within PBMCs are established between high and low responders at the time of vaccination and at six days post-vaccination. A subset of these phosphorylation events were determined to be consistently differentially phosphorylated (p < 0.05) in the validation cohort of high and low vaccine responders. In contrast, there were no differences in birth weight (p > 0.5) and plasma IFNγ concentrations at the time of vaccination (p > 0.6) between high and low responders within the validation cohort. The results in this study suggest, at least within this study population, phosphorylation biomarkers are more robust predictors of vaccine responsiveness than other physiological markers.

Development of an indirect competitive enzyme linked immunosorbent assay for the quantitative detection of Mycoplasma hyopneumoniae during the vaccine production process

Inactivated Mycoplasma hyopneumoniae vaccine is used extensively to control M. hyopneumoniae infection worldwide. Quantification techniques are essential in the process of standardizing and validating vaccines. In this study, we developed and optimized an indirect competitive enzyme linked immunosorbent assay (ic-ELISA) for the rapid quantification of M. hyopneumoniae antigen during vaccine production. Briefly, whole M. hyopneumoniae antigen was coated onto microtiter plates, and a polyclonal antibody against M. hyopneumoniae recombinant elongation factor thermo unstable (EF-Tu) protein was prepared and added with the samples to be tested. The methods were optimized and showed significant reproducibility, with coefficients of variation of 4.01% and 6.14% for the intra-and inter-assays, respectively. Quantification of M. hyopneumoniae cultures at different growth stages using the ic-ELISA test showed a similar curve to that of the traditional color changing units (CCU) assay, with a delay in the time when the amount reached the peak and started to fall. In the inactivated vaccine production process, the cultures could be harvested later than that for the live vaccine, at about 12 h after the end of the logarithmic growth phase. Different batches of cultures were measured for their relative potency value compared with the in-house reference vaccine, which was used to determine whether the cultures met the antigen amount requirements for vaccine preparation. The curves of the CCU titer and ic-ELISA titer in the logarithmic phase correlated strongly and a linear regression equation was established to calculate the CCU values rapidly using the ic-ELISA results. In conclusion, an ic-ELISA method was established to rapidly assess the amount of antigen in an M. hyopneumoniae culture during the vaccine production process.

Cellular and humoral immunity following vaccination with two different PCV2 vaccines (containing PCV2a or PCV2a/PCV2b) and challenge with virulent PCV2d

Porcine Circovirus type 2 (PCV2) associated disease is one of the most economically important swine diseases worldwide. Vaccines reduce PCV2 disease by inducing humoral immunity (neutralizing antibodies) and cell-mediated immunity (CMI) but may be improved by optimizing the immune response they induce. This study evaluated immune responses to a trivalent inactivated Porcine Circovirus (PCV) Type 1-Type 2a chimera (cPCV2a), cPCV2b and Mycoplasma hyopneumoniae (MH) (an experimental serial of Fostera® Gold PCV MH, also marketed as Circomax® Myco) vaccine or a bivalent recombinant PCV2a baculovirus expressed ORF2 capsid plus MH vaccine (Circumvent® PCV-M G2). Treatment Groups (T) received two doses of placebo (T01), one full or two split doses of the trivalent vaccine (T02, T03) or two split doses of the bivalent vaccine (T04) where two doses were given, there was a three-week period between administrations. All pigs were challenged with a virulent field isolate of PCV2d. CMI was measured as PCV2-specific IFN-γ secreting cells in blood and lymph node. Humoral immunity was measured as PCV2 antibodies. Vaccine efficacy was determined as viremia and fecal shedding of virus. There was a robust antibody response in T02 and T04 post the second vaccination and all vaccinated groups post challenge. There was a robust PCV2-specific IFN-γ response following the 1st dose in T02 and T03 and after the second dose in T02. T04 induced a low but detectable PCV2-specific IFN-γ response only after the 2nd dose. Among lymph node cells (study day 52), there was a significantly higher PCV2-specific, IFN-γ response to replicase and PCV2d capsid peptides in T01, consistent with active viral replication in non-vaccinated pigs. The trivalent chimeric vaccine induced robust CMI and protective efficacy, following a one dose regimen or splitting the dose into two vaccine administrations.

Convenience and economic benefit of early one-shot Mycoplasma hyopneumoniae vaccination at 3 days of age in a commercial sow farm

What to do in the event of a varicella infection in pregnant women or children is sometimes not well known. Several therapies are available to doctors; specific immunoglobulins, anti virals, vaccination. We propose a decision tree

Development of an indirect ELISA for detection of anti-Mycoplasma hyopneumoniae IgG in naturally infected pathogen-induced convalescent sera

BackgroundImmunization of pigs with an inactivated Mycoplasma hyopneumoniae vaccine (bacterin) generates hyperimmune serum that contains high concentrations of anti-M. hyopneumoniae IgG. Commercially available IgG-ELISA kits cannot distinguish between anti-M. hyopneumoniae IgG in inactivated bacterin-induced hyperimmune sera and convalescent sera resulting from natural M. hyopneumoniae infection. Establishment of an ELISA to detect anti-M. hyopneumoniae IgG in convalescent sera will facilitate the evaluation of the M. hyopneumoniae status of pig farms.ResultsIn this study, we expressed and purified recombinant Mhp366-N protein, which contains an epitope recognized by M. hyopneumoniae convalescent sera but not hyperimmune sera, for use as a coating antigen. For the M. hyopneumoniae convalescent serum IgG-ELISA, the optimal antigen concentration, blocking buffer, blocking time, dilution of serum, incubation time with serum, secondary antibody dilution, secondary antibody incubation time and colorimetric reaction time were 0.25 µg/mL, 2.5 % skim milk, 1 h, 1:500, 0.5 h, 1:10,000, 1 h and 15 min, respectively. Validation of the M. hyopneumoniae convalescent serum IgG-ELISA showed a cut-off value of 0.323, the intra-assay CV ranged from 3.27 to 7.26 %, the inter-assay CV ranged from 3.46 to 5.93 %, and the assay was able to differentiate convalescent sera from antibodies to 7 other porcine respiratory pathogens. The convalescent serum IgG-ELISA detected no anti-M. hyopneumoniae IgG in hyperimmune serum samples while a commercial IgG-ELISA identified 95/145 of these sera as positive. The accuracy of the M. hyopneumoniae convalescent serum IgG-ELISA was comparable to the sIgA-ELISA but better than the commercial IgG-ELISA.ConclusionsThe convalescent serum IgG-ELISA is a reproducible, sensitive, and specific indirect ELISA to detect anti-M. hyopneumoniae IgG in naturally infected pathogen-induced convalescent sera. This ELISA could be used to carry out large scale surveillance of M. hyopneumoniae infection in pig farms regardless of vaccination status.

Infection status of Mycoplasma hyopneumoniae in experimental pigs at a commercial farm

The objective of this study was to investigate the profiles of Mycoplasma hyopneumoniae (MH) infection at different ages of pig in a sow – finishing herd using serological and molecular methods. A total of 30 study piglets were born from non-vaccinated sows with MH. They were injected one-dose of inactivated MH vaccine at the 10th week. MH infection status was evaluated by using ELISA to detect MH antibodies from blood samples, and PCR to detect MH DNA in nasal swabs or oral fluid samples every other week from newborn to slaughter time. The results of this study showed that PCR positive proportions were low at 1st-2nd week (7-13%), then increased significantly during 5th -7th week (73-79%) and reduced at 8th week (33%); finally became negative after 13th week of age. This pattern corresponds to the one of antibody level. In particular, the level of maternal antibodies against MH was very high due to maternal immunity, then decreased gradually to negative at 7-8 weeks of age, and finally increased gradually from 13 weeks of age to all positive at 25 weeks of age. In conclusion, the result showed that in this herd, MH might invade pigs by the time of 5-7 weeks of age after maternal immunity disappears, and humoral response can overcome the infection at week 13. This should be noted to have appropriate strategies to control MH at the farm.

Clinical efficacy of two vaccination strategies against Mycoplasma hyopneumoniae in a pig herd suffering from respiratory disease

BackgroundA randomised field trial was conducted on an Austrian farrow-to-finish farm for one year to compare the efficacy of two commercial Mycoplasma hyopneumoniae vaccines. 585 piglets either received the one-shot formulation in group 1 (Hyogen®, 23.9 days of age) or a two-shot vaccine (Stellamune® Mycoplasma, 4.3 and 24.0 days of age) in group 2. Assessment of vaccine efficacy was evaluated by regression analyses through cough monitoring from nursery to slaughter, average daily weight gain from inclusion to slaughter, antibiotic treatment rate (ATR), mortality rate, and lung lesion scoring at slaughter.ResultsIn general, coughing was more frequent during late nursery and finishing. No significant differences were found in the coughing index (0.02 vs 0.03) and mean average daily weight gain (560 vs 550 g) between the two groups. ATR was higher in group 2 (3.8 vs 9.6%). At the slaughterhouse check, significant differences in the prevalence of bronchopneumonia (62.9 vs 71.2%) could be found. Extension of lung lesions was also significantly lower in group 1 in terms of enzootic pneumonia (EP) values (p = 0.000, z = − 4.269). There were no significant differences in the rate of scarred lungs (20.0 vs 24.0%) or those affected by dorsocaudal pleurisy (36.8 vs 34.3%).ConclusionsThis trial demonstrated that Hyogen® was superior to Stellamune® Mycoplasma in reducing (I) the prevalence of bronchopneumonic lungs and those affected by cranioventral pleurisy, (II) the extension and severity of EP-like lung lesions, and (III) the rate of antibiotically treated animals against respiratory disease.

Field study on the safety and efficacy of intradermal versus intramuscular vaccination against Mycoplasma hyopneumoniae

The present study compares the safety and efficacy of a needle-free, intradermal Mycoplasma hyopneumoniae vaccine to an intramuscular one. 420 piglets (21+3 days of age) were randomly assigned to two...

Mycoplasma hyopneumoniae vaccination at or shortly before weaning under field conditions: a randomised efficacy trial

This study assessed the efficacy of two different Mycoplasma hyopneumoniae vaccination programmes in relation to the time of weaning. Eight hundred and twenty-eight piglets were randomly divided into three groups:...

Efficacy of Mycoplasma hyopneumoniae vaccination before and at weaning against experimental challenge infection in pigs.

BackgroundCommercial bacterins are widely used at weaning to control Mycoplasma hyopneumoniae infections in pigs. However, it is not known whether the efficacy of vaccinating against M. hyopneumoniae can be influenced by the weaning process when vaccination is applied at the day of weaning. The present study assessed the efficacy of a single M. hyopneumoniae vaccination (Ingelvac MycoFLEX®) three days before weaning (V1) or at weaning (V2) against experimental challenge infection. Four weeks after vaccination, groups V1 and V2 (n = 20 pigs each) and a non-vaccinated, positive control group (PCG) (n = 20) were endotracheally inoculated with a virulent M. hyopneumoniae field strain. Five pigs were used as a negative control group. All pigs were euthanized 5 weeks after challenge. The main parameters investigated included macroscopic and histopathological lung lesions at necropsy, immunofluorescence (IF) staining and quantitative real-time PCR (qPCR) on broncho-alveolar lavage (BAL) fluid for quantifying M. hyopneumoniae.ResultsThe average macroscopic lung lesion scores in groups V1, V2 and PCG were 0.54, 0.88 and 1.04, respectively (P > 0.05). The average lymphohistiocytic infiltration scores in groups V1, V2 and PCG were 2.95, 3.16 and 3.61, respectively (P < 0.05). The average IF scores were: V1 = 1.13, V2 = 1.19 and PCG = 1.25 (P > 0.05), the qPCR values were: V1 = 102.94, V2 = 102.76 and PCG = 103.23 (P > 0.05). All pigs of the negative control group remained negative throughout the study.ConclusionsBoth vaccinated groups had lower numbers of macroscopic and histopathological lung lesions, and lower numbers of M. hyopneumoniae organisms in the BAL fluid compared to the PCG. However, no firm conclusions could be made on whether weaning negatively influences the efficacy of M. hyopneumoniae vaccination, since significant differences between the treatment groups were only obtained for the histopathological lung lesions. This could be attributed to the fact that milder macroscopic lung lesions were produced in the inoculated pigs, when compared to previous trials conducted by the same group. Further research under field conditions is warranted to assess possible differences between the two vaccination strategies.

Efficacy and safety of a new intradermal PCV2 vaccine in pigs

The safety and efficacy of a new intradermal one dose vaccine containing Porcine Circovirus type 2 (PCV2) antigen – Porcilis® PCV ID – was evaluated in laboratory studies and under field conditions. In addition, the concurrent use with an intradermal Mycoplasma hyopneumoniae vaccine – Porcilis® M Hyo ID ONCE – was evaluated.Vaccination with Porcilis® PCV ID resulted in small transient local reactions in a high percentage of the vaccinated animals with no temperature increase. In both the onset of immunity and duration of immunity challenge studies with PCV2 or M. hyopneumoniae, significant reduction of the PCV2 load in lymphoid tissue, lungs, serum and fecal swabs and M. hyopneumoniae-induced lung lesions were observed. In two field trials on two different farms where both PCV2 and M. hyopneumoniae were present, vaccination with Porcilis® PCV ID and/or Porcilis® M Hyo ID ONCE of 3week old piglets resulted in a significant reduction of PCV2 viraemia, mortality and lung lesion scores at slaughter. In addition, a significant positive effect on average daily weight gain (between 44 and 59g/day) in the finishing phase was observed. The results support that this new intradermal vaccine is safe and efficacious against PCV2 and may be used concurrently with Porcilis® M Hyo ID ONCE.

Immunogenic properties of Landrace pigs selected for resistance to mycoplasma pneumonia of swine.

Mycoplasma pneumonia of swine (MPS) lung lesions and immunogenic properties were compared between a Landrace line that was genetically selected for reduced incidence of pulmonary MPS lesions, and a non‐selected Landrace line. The MPS‐selected Landrace line showed significantly lower degrees of pulmonary MPS lesions compared with the non‐selected Landrace line. When changes in immunity before and after vaccination were compared, the percentage of B cells in the peripheral blood of the MPS‐selected Landrace line was significantly lower than that of the non‐selected line. Furthermore, the concentration of growth hormone and the mitogen activity of peripheral blood mononuclear cells in the MPS‐selected Landrace line showed significantly (P < 0.05) lower increases after vaccination than the non‐selected line. Conversely, the concentration of peripheral blood interferon (IFN)‐γ and salivary immunoglobulin A (IgA) after Mycoplasma hyopneumoniae vaccination was significantly higher in the MPS‐selected Landrace line than in the non‐selected line. Gene expression of toll‐like receptor (TLR)2 and TLR4 was significantly higher in the MPS‐selected Landrace line in immune tissues, with the exception of the hilar lymph nodes. The present results suggest that peripheral blood IFN‐γ, salivary IgA TLR2, and TLR4 are important immunological factors influencing the development of MPS lesions.

Characterization of a novel oil-in-water emulsion adjuvant for swine influenza virus and Mycoplasma hyopneumoniae vaccines

Vaccines consisting of subunit or inactivated bacteria/virus and potent adjuvants are widely used to control and prevent infectious diseases. Because inactivated and subunit antigens are often less antigenic than live microbes, a growing need exists for the development of new and improved vaccine adjuvants that can elicit rapid and long-lasting immunity. Here we describe the development and characterization of a novel oil-in-water emulsion, OW-14. OW-14 contains low-cost plant-based emulsifiers and was added to antigen at a ratio of 1:3 with simple hand mixing. OW-14 was stable for prolonged periods of time at temperatures ranging from 4 to 40°C and could be sterilized by autoclaving. Our results showed that OW-14 adjuvanted inactivated swine influenza viruses (SIV; H3N2 and H1N1) and Mycoplasma hyopneumoniae (M. hyo) vaccines could be safely administered to piglets in two doses, three weeks apart. Injection sites were monitored and no adverse reactions were observed. Vaccinated pigs developed high and prolonged antibody titers to both SIV and M. hyo. Interestingly, antibody titers were either comparable or greater than those produced by commercially available FluSure (SIV) or RespiSure (M. hyo) vaccines. We also found that OW-14 can induce high antibody responses in pigs that were vaccinated with a decreased antigen dose. This study provides direct evidence that we have developed an easy-to-use and low-cost emulsion that can act as a powerful adjuvant in two common types of swine vaccines.

Efficacy and safety of a combined Porcine Circovirus and Mycoplasma hyopneumoniae vaccine in finishing pigs

The safety and protective efficacy of a new one dose combination vaccine containing Porcine Circovirus type 2 (PCV2) and M. hyopneumoniae antigens – Porcilis® PCV M Hyo - was evaluated in laboratory studies and under field conditions. Vaccination resulted in a moderate temperature increase on the day of vaccination and mild systemic and local reactions were found in only a low percentage of the vaccinated pigs. The local reactions observed were small (max. 2 cm) and transient (max. 1 day). In short term (onset of immunity) and long term (duration of immunity) challenge studies with the individual pathogens, the vaccine significantly reduced the PCV2 load in lymphoid tissue and lungs and M. hyopneumoniae-induced lung lesions. In a placebo-controlled field trial on a farm where both PCV2 and M. hyopneumoniae were present, vaccination of piglets at 3 weeks of age resulted in a reduction of PCV2 viremia and shedding and lower lung lesion scores at slaughter. In addition, a positive effect on the average daily weight gain (+ 34 g/day) in the finishing phase was observed. It can therefore be concluded that this new ready to use combination vaccine is safe and efficacious against PCV2 and M. hyopneumoniae single and combined infections.

Innate and adaptive immune responses of snatch-farrowed porcine-colostrum-deprived pigs to Mycoplasma hyopneumoniae vaccination.

BackgroundThe snatch-farrowed porcine-colostrum-deprived (SF-pCD) pig model, in which neonates are raised on commercially available bovine colostrum, is an alternative model for porcine infectious disease research. It is not known if SF-pCD pigs possess growth performance and immunity comparable to conventional, farm-raised pigs. The current experiment compared growth performance and immune responses of SF-pCD pigs to their farm-raised siblings following Mycoplasma hyopneumoniae (Mhyo) vaccination. Twelve SF-pCD and 13 farm-raised siblings were vaccinated on day 7 (D7) and D26 of age. Body weights were measured once or twice weekly and average daily gain (ADG) was calculated. Peripheral blood mononuclear cells (PBMC) were isolated on D40. Cytokine secretion from PBMC stimulated with Mhyo antigen or phorbol myristate acetate plus ionomycin (PMA/Iono) was assessed using a multiplexed fluorescent microsphere immunoassay (FMIA). Additionally, interferon gamma (IFNγ) secretion from stimulated PBMC was assessed using ELISPOT. Mhyo IgG titers were measured by an ELISA in D40 sera.ResultsGrowth performance did not differ between groups before weaning, but SF-pCD pigs had higher ADG after weaning. In response to Mhyo stimulation, numbers of IFNγ secreting PBMC and levels of interleukin 8 (IL8) and IL10 in PBMC supernatants were significantly higher in SF-pCD pigs, as were Mhyo antibody levels in sera, and levels of IL1β, IL8 and IL12 in supernatants of PMA/Iono stimulated PBMC.ConclusionsUnder the conditions of this experiment, SF-pCD pigs demonstrated superior growth performance and enhanced humoral and cell-mediated immunity following vaccination. Whether or not this reflects greater resistance or tolerance to infection is unknown but the ability to react positively to the vaccination provides evidence that SF-pCD pigs are a suitable alternative model for swine disease research.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-014-0219-2) contains supplementary material, which is available to authorized users.

A field evaluation of two vaccines against Mycoplasma hyopneumoniae infection in pigs.

BackgroundA field trial was carried out with two Mycoplasma hyopneumoniae vaccines in order to investigate the benefit of vaccination under field conditions in modern Danish pig production facilities with pigs being positive for M. hyopneumoniae. The M. hyopneumoniae infection of the herd was confirmed through blood samples that were positive for antibodies against M. hyopneumoniae combined with gross lesions of the lungs related to M. hyopneumoniae at slaughter and detection of M. hyopneumoniae by polymerace chain reaction in these lesions.ResultsA total of 2,256 pigs from two herds were randomly divided into three groups. Group 1 received 2 mL ThoroVAX®VET, Group 2 received 1 mL Ingelvac®MycoFLEX, and Group 3 was a non-vaccinated control group. The vaccination was performed by a person who was not involved in the rest of the trial and vaccination status thereby blinded to the evaluators.The prevalence of lung lesions related to M. hyopneumoniae were significantly lower for pigs vaccinated with ThoroVAX®VET but not for pigs vaccinated with Ingelvac®MycoFLEX®, when compared to non-vaccinated pigs. There was no significant effect of vaccination on growth rate, antibiotic consumption or mortality.ConclusionThis trial demonstrated that vaccination with Thoro®VAX VET was effective in reducing the prevalence of lung lesion in pig units infected with M. hyopneumoniae.

Effect of different adjuvant formulations on the immunogenicity and protective effect of a live Mycoplasma hyopneumoniae vaccine after intramuscular inoculation

Mycoplasma hyopneumoniae (M. hyopneumoniae) vaccine strain 168 is an intrapulmonically injected attenuated live vaccine that is available in the Chinese market. The aim of this study was to develop suitable adjuvants for this live vaccine to provide effective protection after intramuscular inoculation. Several adjuvant components were screened to assess their toxicity for the live vaccine, and various adjuvant formulations were then designed and prepared. Vaccines supplemented with these adjuvants were used to immunize mice intramuscularly to assess the capacity of the adjuvants to induce a specific immune response. The screened formulations were then evaluated in pigs. Seven of the eight adjuvant components did not affect the viability of the live vaccine, and seven different adjuvant formulations were then designed. In mice, the ISCOM-matrix adjuvant and the levamisole–chitosan mixture adjuvant significantly enhanced serum IgG responses against M. hyopneumoniae, while lymphocyte proliferation was enhanced by the ISCOM-matrix adjuvant, the carbomer–astragalus polysaccharide mixture adjuvant and an oil-in-water emulsion adjuvant. These four adjuvants were evaluated in pigs. Enhancement of specific lymphocyte proliferation responses was observed in the groups vaccinated with the ISCOM-matrix adjuvant and the carbomer–astragalus polysaccharide mixture adjuvant. Significant enhancement of serum IgG antibody production was observed before challenge in pigs vaccinated with the carbomer–astragalus polysaccharide mixture adjuvant and the levamisole–chitosan mixture adjuvant, while after challenge, all of the animals that received vaccines containing adjuvants had higher antibody concentrations against M. hyopneumoniae than unvaccinated animals. Animals inoculated with a vaccine containing the ISCOM-matrix adjuvant (median score 3.57) or the carbomer–astragalus polysaccharide mixture adjuvant (median score 5.28) had reduced lesion scores compared to unvaccinated animals (median score 14.81). These studies will help in the development of appropriate adjuvants for intramuscular administration of this live M. hyopneumoniae vaccine.

Interaction of porcine circovirus type 2 and Mycoplasma hyopneumoniae vaccines on dually infected pigs

The objective of this study was to determine the effects of porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae vaccinations on disease severity in an experimental PCV2-M. hyopneumoniae dual challenge model. Vaccine effectiveness was evaluated using microbiological (PCV2 viremia and M. hyopneumoniae nasal shedding), immunological (neutralizing antibodies and interferon-γ-secreting cells), and pathological (gross lung lesions, histopathologic pulmonary and lymphoid lesions, and the presence of PCV2 antigen and M. hyopneumoniae DNA within the lesions) evaluations. Although M. hyopneumoniae potentiates the severity of PCV2-associated lesions and lesion-associated PCV2 antigen in dually challenged pigs, vaccination against M. hyopneumoniae alone did not reduce PCV2 viremia, PCV2-induced lesions, or PCV2 antigen in dually challenged pigs. In addition, vaccination against PCV2 did not reduce the nasal shedding of M. hyopneumoniae, the M. hyopneumoniae-induced pulmonary lesions or the lesion-associated M. hyopneumoniae DNA in dually challenged pigs. Dual challenge with PCV2 and M. hyopneumoniae did not interfere with the induction of active immunity induced by a previous single vaccination for either PCV2 or M. hyopneumoniae. The results of this study demonstrated that (i) vaccination against M. hyopneumoniae alone did not decrease the potentiation of PCV2-induced lesions by M. hyopneumoniae and (ii) vaccination against PCV2 alone decreased the potentiation of PCV2-induced lesions by M. hyopneumoniae in dually challenged pigs.