Mycoplasma hyopneumoniae (M. hyopneumoniae) is the primary agent of enzootic pneumonia in pigs. Pigs are often infected with different M. hyopneumoniae strains. This study assessed the efficacy of vaccination against experimental infection with two genetically different M. hyopneumoniae strains in weaned piglets. At 33 days of age (D0), 45 M. hyopneumoniae-free piglets were randomly assigned to three different groups: 1) negative control group (NCG; n = 5): not vaccinated, not infected, 2) positive control group (PCG; n = 20): not vaccinated, infected, and 3) vaccination group (VG; n = 20): single vaccination with an inactivated whole-cell M. hyopneumoniae vaccine (Hyogen®, Ceva) (D1), infected. The PCG and VG were endotracheally inoculated with 7 × 107 CCU in 7 ml of the highly virulent M. hyopneumoniae strain F7.2C (D24) and 7 × 107 CCU in 7 ml low virulent strain F1.12A (D25). A respiratory disease score (RDS) was assessed from D24 until D53. At D53 (euthanasia), macroscopic lung lesions (MLL) were scored, log copies of M. hyopneumoniae DNA (qPCR) and IL-1 and IL-6-concentrations (ELISA) on bronchoalveolar lavage fluid were determined. The RDS and MLL at euthanasia were respectively 0, 1.20 and 0.55 (P < 0.001) and 0, 7.56 and 0.68 (P < 0.001) for NCG, PCG and VG, respectively. The qPCR results for PCG and VG were 3.99 and 1.78 log copies (P < 0.001), respectively, with a significant difference between PCG and VG. The IL-1 and IL-6 results at euthanasia for NCG, PCG and VG were 17.61, 1283.39 and 53.04 pg/ml (P < 0.001) and 148.10, 493.35 and 259.80 pg/ml (P = 0.004), respectively with a significant difference between PCG and VG. Vaccination with Hyogen® in pigs was efficacious against an experimental challenge with both a low and highly virulent M. hyopneumoniae strain as the vaccinated pigs coughed significantly less, and showed significantly less lung lesions compared to the non-vaccinated challenged pigs: the vaccinated animals showed a 52.9% lower RDS and 91.0% lower MLL compared to the PCG. In the bronchoalveolar lavage fluid collected at the necropsy of the vaccinated pigs, a significantly lower amount of M. hyopneumoniae-DNA and a significantly lower IL-1 and IL-6 concentration was found compared to the pigs of the PCG.

This study aimed to describe Mycoplasma hyopneumoniae (M. hyopneumoniae) genetic variability in vaccinated (V) and non-vaccinated (NV) slaughtered pigs showing cranio-ventral pulmonary consolidation (CVPC). Ten V and 10 NV fattening farms with respiratory problems associated to M. hyopneumoniae were selected. Lung lesions of one batch per farm were scored at slaughterhouse and the enzootic pneumonia (EP)-index was calculated. Moreover, three lungs showing the most extensive CVPC per farm were sampled and tested for M. hyopneumoniae detection by real-time (rt)-PCR. Positive samples with cycle threshold <= 30 were selected to be genotyped by sequencing of four loci (P97, P146, H1 and H5). Typing profiles (TP) were assigned considering four or two (P97, P146) loci. Five commercial vaccines for M. hyopneumoniae (VS) and two reference strains (RF) were also genotyped. The EP-index (mean +/- SD) in NV farms (3.8 +/- 1.9) was not significantly different from V ones (2.2 +/- 1.3). From the 60 selected lungs, 46 (76.7%) were M. hyopneumoniae positive by rt-PCR (25/30 and 21/30 from NV and V farms, respectively), and 43 (93.5%) of those were successfully genotyped. A total of 24 different TP(12 in V and 12 in NV farms) or 17 TP(9 in V and 9 in NV farms, being one TP in both farm types) were identified by analyzing four or two loci, respectively. One to three TP per farm were detected, being different from VS and RF. Interestingly, farms with same breeding origin had the same TP using two loci, but such link was not found using four loci. Therefore, high inter-farm and limited intra-farm M. hyopneumoniae genetic variability were detected, but variability depended on the number of studied loci.

Piglets from a porcine circovirus type 2 (PCV2) stable farm of low and high levels of maternally derived antibodies (MDA) against PCV2 were vaccinated either with a whole virus type or a PCV2 ORF2 antigen-based commercial subunit vaccine at three weeks of age. Two non-vaccinated groups served as low and high MDA positive controls. At four weeks post vaccination, all piglets were challenged with a PCV2d-2 type virus strain and were checked for parameters related to vaccine protection over a four-week observation period. MDA levels evidently impacted the outcome of the PCV2d-2 challenge in non-vaccinated animals, while it did not have a significant effect on vaccine-induced protection levels. The humoral immune response developed faster in the whole virus vaccinates than in the subunit vaccinated pigs in the low MDA groups. Further, high MDA levels elicited a stronger negative effect on the vaccine-induced humoral immune response for the subunit vaccine than for the whole virus vaccine. The group-based oral fluid samples and the group mean viraemia and faecal shedding data correlated well, enabling this simple, and animal welfare-friendly sampling method for the evaluation of the PCV2 viral load status of these nursery piglets.

Vertical transmission is a consistently discussed pathway of porcine circovirus type 2 (PCV2) and porcine circovirus type 3 (PCV3) transmission in pigs. To evaluate the presence of PCV2 and PCV3 in piglets, we collected tissue samples from 185 piglets that were crushed within the first week of life from 16 farms located in Germany and Austria. Pooled samples consisting of thymus, inguinal lymph node, myocardium, lung and spleen were examined for PCV2 and PCV3 by qPCR. Furthermore, oral fluid samples (OFS) from grow–finish pigs were collected and examined the same way. In piglets, PCV2 was highly prevalent (litters: 69.4%; piglets: 61.6%), whereas PCV3 prevalence was low (litters: 13.4%; piglets: 13.0%). In total, 72.6% and 67.2% of all collected OFS were PCV2 or PCV3 positive, respectively. Sow vaccination against PCV2 was identified as a protective factor concerning PCV2 in piglets (OR: 0.279; CI: 0.134–0.578; p < 0.001), whereas the porcine reproductive and respiratory syndrome virus (PRRSV) vaccination of sows was identified as a protective factor concerning PCV3 in piglets (OR: 0.252 CI: 0.104–0.610; p = 0.002). Our results show that PCV2, but not PCV3, is ubiquitous in suckling piglets and that early PCV3 infections might be modulated by PRRSV–PCV3 interaction. However, the ubiquitous nature of both viruses in older pigs could be confirmed.

This study evaluated different gilt vaccination protocols against Mycoplasma (M.) hyopneumoniae at acclimation and their effect on the genetic diversity. A total of 180 M. hyopneumoniae naïve gilts were selected 1 week post-entry (wpe) at the acclimation barn in a clinically affected M. hyopneumoniae farm. Gilts were distributed according to the M. hyopneumoniae antibodies levels into three different vaccination schedules: A) four doses of a M. hyopneumoniae commercial vaccine at 2, 4, 6 and 8 wpe; B) two vaccine doses at 2 and 6 wpe and PBS at 4 and 8 wpe; and C) four PBS doses at the same wpe. Detection of M. hyopneumoniae (rt-PCR) and antibodies (ELISA) were assessed in gilts at 1, 14, 27 and 34 wpe and in 6 of their piglets at weaning. Rt-PCR positive gilts were detected at 14 wpe, being the proportion significantly lower in groups A and B (3/120, 3%) than C (27/60, 45%). Seroconversion was detected at 14 wpe, showing significant differences in percentage of inhibition (PI) between groups A (median 4.9, range 3.1-19.9) and B (5.5, 3.7-13.5), and C (14.3, 3.3-53.2). Gilts remained seropositive over the study and significant differences in PI were detected between groups A and B versus C. All piglets were rt-PCR negative, but the proportion of seropositive piglets coming from vaccinated gilts was significantly higher than the non-vaccinated group. M. hyopneumoniae characterization showed high variability. Hence, gilt vaccination with 2 or 4 doses significantly decreased the pathogen infectious pressure, variability, and provided high antibody levels to gilts and their offspring.

Mycoplasma hyopneumoniae (M.?hyo) and porcine circovirus type?2 (PCV-2) are major and widespread swine pathogens, both implicated in?the porcine respiratory disease complex, which can lead to?significant economic losses for pig producers. PCV-...