Early detection of respiratory distress is essential to preserve animal health and to combat economic losses due to infections with respiratory pathogens. Within the present study we combined a sound based A.I. respiratory health monitoring system with a molecular biological laboratory screening based on OFs and AS for the detection of pathogens associated with PRDC. Within this scope, we also compared OFs and AS as matrices for the detection of nucleic acids of PRRSV, PCV2, APP and swIAV. As observed by others [
41] the A.I. based monitoring system proved to be technically reliable and respiratory health data could be obtained for each study day. However, limitations concerning the use of such a monitoring might display the lack of internet within the range of the stables, which should be checked prior to the installation of such a monitoring system. The comparison between the 24/7 A.I. and manual coughing monitoring correlated on a moderate to medium level, as also reported by others [
16]. In principle this correlation was expected as coughing is needed to have a corresponding response by the A.I. monitoring system. However, the ReHS gave more detailed information, e.g., in the first batch, when coughing was present particularly in the front part of nursery unit. Thus, next to the recognition of clinical signs, the A.I-based long-time monitoring on farms might also demonstrate localized pattern of disease that indicate issues concerning the farm environment or internal biosecurity. The advantage of continuous A.I based monitoring becomes clearly recognizable as different times of activity of pigs over a day period can be addressed, whereas a clinical examination is limited in time and only covers the time of physical presence in the corresponding pig population. This might explain the only moderate correlation between ReHS and CCS in the present examination. The circadian rhythm of the pig in its surrounding environment influences the times of activity of the pigs [
42] and thus, the appearance of clinical signs due to the circadian oscillation of the immune system. In terms of respiratory diseases in humans, the severity of clinical signs also shows circadian variability across the 24-h cycle. An increased inflammation and disease severity at night is described for obstructive airways diseases and allergic rhinitis with the consequence of greater effects to exposure of inflammatory insults at night [
24]. Based on a previous investigation the optimal number of devices for the 600 head barn was determined to be two [
43]. However, to ensure optimal sound coverage and to account for variations in infection dynamics the number of required monitors and the placement of the devices must be thoroughly considered.
The diagnostic screening by PCR revealed the presence of multiple respiratory pathogens in the nursery of the study farm. Thus, the principal preconditions for PRDC were present in the study population. Nevertheless, in order to definitively determine whether a certain combination of pathogens contributed to the clinical signs, pathomorphological examinations would have been required. Interestingly, a significant correlation was observed between decreasing Ct-values of the swIAV PCR and increasing respiratory distress measured by the A.I. or the investigator. Thus, an increase of the swIAV viral load in OFs or AS coincided with the extend of the clinical signs in terms of coughing expressed by reduced ReHS or increased CCS, respectively. Comparable observations were made by Neira, et al. [
44] who reported a correlation between the quantitative detection of swIAV in oral fluids and the coughing score. Besides the link between swIAV viral load in OFs or As and the extend of coughing, this observation also gives evidence to suggest that swIAV might have been the driving force for respiratory distress in this nursery unit. Moreover, based on the comparable pattern of swIAV RNA detection in both batches an endemic swIAV infection can be assumed. As already postulated by Prost, et al. [
35] and Anderson, et al. [
34], no significant differences concerning the qualitative detection of swIAV by PCR were evident between OFs and bioaerosol samples. However, in contrast to Prost, et al. [
35] swIAV RNA loads were significantly higher in OFs compared to bioaerosol samples. The detection of PRRSV might be a result of the MLV vaccination of the piglets as no significant correlation between clinical observations and PRRSV detection was observed in the present study. However, no further diagnostics to discriminate between field or vaccine strains could be conducted due to the low viral loads in both sampling materials. Shedding of MLVs is a well-known observation after vaccination and viral RNA could be detected in nasal swabs by PCR under experimental conditions up to 42 days after vaccination with an MLV [
45]. Although airborne transmission of PRRSV over more than four kilometers was reported [
36], the detection rate of viral nucleic acids in bioaerosol samples compared to OFs in our study was rather low. This observation indicates less suitability of bioaerosols for monitoring or surveillance purposes of PRRSV compared to OFs whose sensitivity and suitability to monitor PRRSV was already shown elsewhere [
46,
47,
48]. Still, these results should be interpreted with caution because airborne transmission of PRRSV might be strain depended as reported by others [
37] and the attenuation of the vaccine strain that was putatively detected here might bias our findings. Significant advantages of OFs over AS could also be observed for the detection of
A. pleuropneumoniae. The high sensitivity of OFs concerning the detection of
A. pleuropneumoniae by PCR was already shown in a previous study under field conditions [
27]. The sporadic detection of
A. pleuropneumoniae DNA in the bioaerosols is in line with the limited capability of
A. pleuropneumoniae aerosol transmission reported elsewhere [
49] and the higher within pen transmission rate compared to the transmission between different pens [
50]. However, the sporadic detection of APP in the bioaerosols indicates the possibility of airborne infections with
A. pleuropneumoniae within a pig population as shown by others [
49,
50]. Moreover, differences in the ventilation rate, air humidity, particle size and the overall bacterial load in the pig population might influence the detection of bacteria in bioaerosol samples. Particularly the underfloor extraction for the exhaust air of the stable in the present study might contribute to a reduced the number of bacteria in the air, which could also in a certain extend apply to
A. pleuropneumoniae. Concerning PCV2, Harms, et al. [
8] demonstrated the relevance of the combination of PCV2 and swIAV in a PRDC case series. However, in the present study the qualitative and quantitative detection rate of PCV2 was low in both sample types. Although Nielsen, et al. [
51] postulated that PCV2 detection in OF are not necessarily correlated with clinical signs, PCV2 does not seem to play a major role concerning the clinical outcome in the present nursery, as evident from
Figure 5. Moreover, the pigs of the present examination were vaccinated against PCV2. The positive effects of PCV2 vaccination concerning PRDC or PMWS affected herds [
52,
53] on clinical outcomes and co-infections are widely known.