Aims
A new multiplex qPCR, targeting Mycoplasma (M.) hyopneumoniae, M. hyorhinis and M. flocculare, was developed and the relationship between detection of those mycoplasma species and the extent of ...gross pneumonia‐like lesions in slaughtered pigs lungs were investigated.
Methods and Results
The multiplex qPCR method targets the p102, p37 and fruA genes and has detection limits of 14, 146 and 16 genome equivalents μl−1 for M. hyopneumoniae, M. hyorhinis and M. flocculare, respectively. In all, 671 lungs were collected and analysed, among them 666 were scored for macroscopic pneumonia and categorized according to the extent of the lesions (no or minor lesions, moderate lesions and extensive lesions). According to results of multiplex qPCR, 59·5% were positive for M. hyopneumoniae, 3·4% for M. hyorhinis and 34·7% for M. flocculare, with on average, 3·1 × 107, 9·7 × 106 and 5·7 × 106 genome equivalents of mycoplasma ml−1, respectively. More results showed that no or minor lesions were associated with multiplex qPCR‐negative results or qPCR‐positive results for M. flocculare. Moderate to extensive lesions were positively correlated with qPCR‐positive results for M. hyopneumoniae. Extensive lesions were associated with qPCR‐positive results for at least two mycoplasma species (M. hyopneumoniae and M. hyorhinis).
Conclusion
The findings also indicated that M. hyopneumoniae and M. hyorhinis significantly increased the odds for a lung to have macroscopic pneumonia. No relationship was found between the extent of lesions and the mycoplasma genome load.
Significance and Impact of the study
This new multiplex qPCR appears to be specific, sufficiently sensitive and repeatable. The validation of this method with field samples guarantees its use for field epidemiological investigations, particularly to gain more insight into the aetiology of the porcine respiratory disease complex.
A study was carried out in 125 farrow-to-finish pig herds to assess the relationships between pathogens involved in respiratory disorders and to relate these findings to clinical signs of respiratory ...diseases and pneumonia and pleuritis at slaughter. Clinical examination and sampling were carried out on four different batches in each herd (pigs aged 4, 10, 16 and 22 weeks). Mycoplasma hyopneumoniae, Actinobacillus pleuropneumoniae, swine influenza viruses (SIV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) were detected by serological or PCR tests. Pneumonia-like gross lesions and pleuritis were scored at the slaughterhouse. The results indicate that the percentage of pigs PCR-positive for PCV2 at 4, 10 and 16 weeks old was associated with the percentage of pigs PCR-positive for M. hyopneumoniae at these ages. On the other hand, the percentage of pigs with antibodies against PRRSV at 10, 16 and 22 weeks was positively correlated with the percentage of pigs seropositive for M. hyopneumoniae at 22 weeks, with the percentage of pigs with antibodies against SIV H1N1 and SIV H1N2 and the percentage of pigs sero-positive for A. pleuropneumoniae serotype 2. The findings also indicate that, within the five studied pathogens, M. hyopneumoniae, PRRSV and SIV H1N1 are the major pathogens involved in pneumonia-like gross lesions even though PCV2 may play a role. A. pleuropneumoniae serotype 2, in association with PRRSV, is significantly associated with extensive pleuritis. Respiratory diseases could be significantly reduced by implementing measures including appropriate management practices to control these pathogens.
•Piglets without MDAs showed an immune response in the 4 weeks post-vaccination.•No immune response was detected for 4 weeks post-vaccination in piglets with MDAs.•MDAs (neutralizing antibody) can ...impair PRRS post-vaccination immune responses in piglets.
The influence of maternally-derived antibodies (MDAs) on the post-vaccination humoral and cellular immune responses in piglets vaccinated against PRRS was studied. The piglets came from a vaccinated breeding herd. Thirty piglets with a low (A−) or high level (A+) of PRRSV-neutralizing MDAs were vaccinated (V+) with a modified live vaccine at 3 weeks of age. Blood samples were collected before vaccination and then at 2, 4, 8 and 14 weeks post-vaccination (WPV). The samples were analysed to detect the vaccine viraemia (RT-PCR) and quantify the post-vaccination humoral (ELISA and virus neutralisation test) and cellular (ELISPOT IFNγ) immune responses. PRRSV vaccine strain was detected in 60%, 64%, 36% and 0% of A−V+ piglets 2, 4, 8 and 14 WPV respectively. No virus was detected in A+V+ piglets during the first four WPV but 32% and 6% of A+V+ piglets were PCR-positive at 8 and 14 WPV. Eighty-five percent of A-V+ piglets and 0% of A+V+ piglets seroconverted (ELISA) between 2 and 4 WPV. Neutralising antibodies appeared 4 WPV in the A-V+ piglets and 14 WPV in the A+V+ piglets. The number of PRRSV-specific IFNγ-secreting cells was significantly higher in A−V+ piglets at 2 and 4 WPV than in A+V+ piglets. These results show that MDAs can affect both post-vaccination humoral and cellular immune responses in piglets. Further studies are required to assess the impact of MDAs on vaccine efficacy following a PRRSV challenge and its ability to reduce viral transmission.
Four sampling techniques for
Mycoplasma hyopneumoniae detection, namely nasal swabbing, oral-pharyngeal brushing, tracheo-bronchial swabbing and tracheo-bronchial washing, were compared in naturally ...infected live pigs. In addition, a quantitative real-time PCR assay for
M. hyopneumoniae quantification was validated with the same samples. 60 finishing pigs were randomly selected from a batch of contemporary pigs on a farm chronically affected by respiratory disorders. Each pig was submitted to nasal swabbing, oral-pharyngeal brushing, tracheo-bronchial swabbing and tracheo-bronchial washing. Nested-PCR and real-time PCR assays were performed on all samples. A Bayesian approach was used to analyze the nested-PCR results of the four sampling methods (
i.e. positive or negative) to estimate the sensitivity and specificity of each method.
M. hyopneumoniae was detected by nested-PCR in at least one sample from 70% of the pigs. The most sensitive sampling methods for detecting
M. hyopneumoniae in live naturally infected pigs were tracheo-bronchial swabbing and tracheo-bronchial washing, as compared to oral-pharyngeal brushing and nasal swabbing. Swabbing the nasal cavities appeared to be the least sensitive method. Significantly higher amounts of
M. hyopneumoniae DNA were found at the sites of tracheo-bronchial sampling than in the nasal cavities or at the oral-pharyngeal site (
p
<
0.001). There was no difference between the tracheo-bronchial washing and the tracheo-bronchial swabbing results (
p
>
0.05). Our study indicated that tracheo-bronchial swabbing associated with real-time PCR could be an accurate diagnostic tool for assessing infection dynamics in pig herds.
•Biosecurity measures reduce PRRSV probability of introduction.•Practices favoring contacts between animals from different batches increase PRRSV infection.•Factors dealing with the control of indoor ...climate are related to PRRSV infection characteristics.
Factors associated with porcine reproductive and respiratory syndrome virus (PRRSV) infection were investigated in 109 herds. Serums from four batches of pigs (4, 10, 16 and 22 weeks, 15 pigs/batch) were tested by ELISA for PRRSV antibodies. Infection by Mycoplasma hyopneumoniae (Mhp), Actinobacillus pleuropneumoniae, H1N1 and H1N2 swine influenza A viruses (swIAV) and PCV2 were detected by specific serological or PCR tests. Data related to herd characteristics, biosecurity, management housing and climatic conditions were collected during a herd visit. Factors associated with the herd’s PRRSV seropositive status were identified by logistic regression. Large herd size, the lack of disinsectisation in the gestation facilities, on-farm semen collection, a short time-period for gilt quarantine and a low temperature setpoint for the ventilation controller in the fattening room significantly increased the odds of a herd being seropositive for PRRSV. Infection by Mhp and H1N2 swIAV were associated with a PRRSV seropositive status. A Cox proportional hazards model was used to identify the factors associated with the age-time to seroconversion in infected herds. Joint housing for the gilts and sows when lactating, a large nursery pen, a small number of pens per fattening room and lack of all-in all-out management in the fattening section significantly reduced the age-time to seroconversion. A small range of temperatures controlling ventilation rate in the nursery room was also associated with time to PRRSV seroconversion. Infection by Mhp and a high PCV2 infection pressure were associated with a shorter time to seroconversion. Biosecurity measures minimising the risk of introducing PRRSV into the herd, management practices reducing contacts between animals from different batches and within batches and favourable climatic conditions should be implemented to better control PRRSV infection.
Infection by Toxoplasma gondii postnatally can occur after ingestion of contaminated meat or water (tissue cysts/oocysts). In Europe, percentage of meat borne infections is estimated between 30 and ...63 %, out of which pork makes the most important source. The aim of this study was to (i) investigate the seroprevalence of T. gondii in intensive pig farms from western France; and (ii) identify the risk factors associated with seropositivity.
Data were collected between November 2006 and February 2008 in 60 intensive farrow-to-finish farms, where sera were taken from 3595 fattening pigs, weaned and suckling piglets. Information about three classes of potential seropositivity risk factors were obtained through a questionnaire concerning: (i) breeding characteristics; (ii) farm management; and (iii) husbandry and hygiene. The modified agglutination test (MAT) was used for detection of specific anti T. gondii antibodies in pig sera, starting from 1/6 dilution.
The overall proportion of seropositive animals was 6.9 %, but the proportion of herds with at least one positive pig was 100 %. Multivariate logistic mixed model showed an increased seropositivity risk in weaned compared to suckling piglets, and a decreasing risk for mid-sized and large farms. The presence of a Danish entry facility, that clearly separates clean and dirty areas, had a protective effect on T. gondii seropositivity as well.
The observed proportion of herds with at least one T. gondii seropositive animal provides further evidence that even in confined conditions of pig breeding, infection occurs, and is common. The highest risk for acquiring T. gondii is at the end of weaning period. Smaller confined pig farms demonstrate higher T. gondii seropositivity levels. This study also showed that Danish entry on farm buildings provides effective protection against T. gondii.
•The commercial SERELISA®PCV2 Ab Mono Blocking kit was compared to an in-house ELISA.•Their diagnostic characteristics were assessed without a gold standard.•465 serum samples from finishing pigs not ...vaccinated against PCV2 were used.•Both ELISAs appeared to be valuable tools for detecting PCV2 antibodies.
This study was designed to assess the diagnostic characteristics of two PCV2 ELISAs without a gold standard. Four hundred and sixty-five serum samples from finishing pigs (25 herds) not vaccinated against PCV2 were used. Samples were tested by two ELISAs: an in-house ELISA (I-ELISA) and the commercial SERELISA®PCV2 Ab Mono Blocking kit (S-ELISA). A ROC curve was used to assess the S-ELISA’s optimal threshold by taking the I-ELISA as a reference and using the cut-off previously determined by comparison to an cccmonolayer assay (IPMA). This led to an S-ELISA result ≥170 being considered as positive. The sensitivity (Se) and specificity (Sp) of each ELISA were then estimated without a gold standard using a Bayesian approach. The mean Se and Sp values of the I-ELISA were slightly higher than those of the S-ELISA (mean Se I-ELISA=0.90 vs. mean Se S-ELISA=0.86; mean Sp I-ELISA=0.92 vs. mean Sp S-ELISA=0.85). However, the 95% credibility intervals (CI95%) overlapped (Se I-ELISA CI95%=0.85–0.95 vs. Se S-ELISA CI95%=0.82–0.90; Sp I-ELISA CI95%=0.82–0.98 vs. Sp S-ELISA CI95%=0.75–0.94). Both ELISAs appeared to be valuable tools for detecting PCV2 antibodies.
Abstract
Background
Up to now, information on the levels of maternally-derived antibodies (MDA) against PCV-2 in suckling piglets born to sows vaccinated with different strategies is scarce in the ...literature. In the present observational study, the PCV-2-specific MDA titres from piglets from 109 farms (thirty 3-day-old and thirty 21-day-old piglets per farm) across four different European countries (France n = 30, Germany n = 27, Italy n = 22 and Spain n = 30) using different sow vaccination strategies (during gestation, as a gilt, as a piglet or never) were assessed.
Results
In all four countries, mean log PCV-2 MDA titres were higher in 3-day-old piglets than in the 3-week-old ones, being significant in most of all the comparisons performed. Within each country, the highest PCV-2-specific MDA titres were observed in the 3-day-old piglets born to sows vaccinated during gestation. Indeed, in the four countries, more than 60% of this subpopulation (3-day-old piglets from sows vaccinated during pregnancy) had the highest log PCV-2 titres detectable with the ELISA technique used in this study. The lowest MDA titres were more variable. Whereas in France and Germany the lowest titres corresponded to 21-day-old piglets born from sows vaccinated as a piglet, in Italy, they corresponded to 21-day-old piglets derived from sows vaccinated as a gilt and in Spain to 21-day-old piglets born from non-vaccinated sows. In this study, PCV-2-specific MDA titres at 3 and 21 days of age were not affected by sow parity.
Conclusions
Data obtained could be considered as a European global overview of PCV-2-specific MDA titres present in the pre-vaccinated piglet populations in different European countries, with titres tending to be higher in younger piglets, but with values variable among countries and sow vaccination strategies.
•Field strain or vaccine immunization partially protect against emerging Lena strain.•Vaccines of genotype 1 or 2 give similar protection against the Lena strain.•Protection induced by field strain ...is slightly better than with vaccine strains.•Cell-mediated immune response contributes to the control of Lena viremia.
The porcine reproductive and respiratory syndrome virus (PRRSV) causes huge economic losses for the swine industry worldwide. In the past several years, highly pathogenic strains that lead to even greater losses have emerged. For the Western European swine industry, one threat is the possible introduction of Eastern European PRRSV strains (example Lena genotype 1.3) which were shown to be more virulent than common Western resident strains under experimental conditions. To prepare for the possible emergence of this strain in Western Europe, we immunized piglets with a Western European PRRSV field strain (Finistere: Fini, genotype 1.1), a new genotype 1 commercial modified live virus (MLV) vaccine (MLV1) or a genotype 2 commercial MLV vaccine (MLV2) to evaluate and compare the level of protection that these strains conferred upon challenge with the Lena strain 4 weeks later. Results show that immunization with Fini, MLV1 or MLV2 strains shortened the Lena-induced hyperthermia. In the Fini group, a positive effect was also demonstrated in growth performance. The level of Lena viremia was reduced for all immunized groups (significantly so for Fini and MLV2). This reduction in Lena viremia was correlated with the level of Lena-specific IFNγ-secreting cells. In conclusion, we showed that a commercial MLV vaccine of genotype 1 or 2, as well as a field strain of genotype 1.1 may provide partial clinical and virological protection upon challenge with the Lena strain. The cross-protection induced by these immunizing strains was not related with the level of genetic similarity to the Lena strain. The slightly higher level of protection established with the field strain is attributed to a better cell-mediated immune response.
Growing and finishing performances of pigs strongly influence farm efficiency and profitability. The performances of the pigs rely on the herd health status and also on several non-infectious ...factors. Many recommendations for the improvement of the technical performances of a herd are based on the results of studies assessing the effect of one or a limited number of infections or environmental factors. Few studies investigated jointly the influence of both type of factors on swine herd performances. This work aimed at identifying infectious and non-infectious factors associated with the growing and finishing performances of 41 French swine herds.
Two groups of herds were identified using a clustering analysis: a cluster of 24 herds with the highest technical performance values (mean average daily gain = 781.1 g/day +/- 26.3; mean feed conversion ratio = 2.5 kg/kg +/- 0.1; mean mortality rate = 4.1% +/- 0.9; and mean carcass slaughter weight = 121.2 kg +/- 5.2) and a cluster of 17 herds with the lowest performance values (mean average daily gain =715.8 g/day +/- 26.5; mean feed conversion ratio = 2.6 kg/kg +/- 0.1; mean mortality rate = 6.8% +/- 2.0; and mean carcass slaughter weight = 117.7 kg +/- 3.6). Multiple correspondence analysis was used to identify factors associated with the level of technical performance. Infection with the porcine reproductive and respiratory syndrome virus and the porcine circovirus type 2 were infectious factors associated with the cluster having the lowest performance values. This cluster also featured farrow-to-finish type herds, a short interval between successive batches of pigs (≤3 weeks) and mixing of pigs from different batches in the growing or/and finishing steps. Inconsistency between nursery and fattening building management was another factor associated with the low-performance cluster. The odds of a herd showing low growing-finishing performance was significantly increased when infected by PRRS virus in the growing-finishing steps (OR = 8.8, 95% confidence interval 95% CI: 1.8-41.7) and belonging to a farrow-to-finish type herd (OR = 5.1, 95% CI = 1.1-23.8).
Herd management and viral infections significantly influenced the performance levels of the swine herds included in this study.
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