Respiratory diseases are considered one of the most important problems in swine production worldwide due to the significant economic losses associated. Lung lesion evaluation at slaughterhouses by different scoring systems is commonly used to monitor respiratory diseases in swine. Concretely, cranioventral pulmonary consolidation lesions are associated with enzootic pneumonia (EP) caused by Mycoplasma hyopneumoniae (Mhyo); whereas haemorrhagic necrotizing pneumonia, mainly in the dorso-caudal lung lobes, and chronic pleuritis (CP) are associated with Actinobacillus pleuropneumoniae (App). Despite the recent consideration of several statistical methods for modelling the temporal dynamics of diseases and the construction of monitoring systems, none have been applied to lung lesions data collected at slaughterhouses. Thus, this work aimed (1) to describe the temporal patterns of EP and CP-like lesions in Spain using time series methods to model the collected data on lung lesions at slaughterhouses; and (2) to construct and evaluate in quasi-real time a surveillance system for early detection of outbreaks and abnormal trends potentially related to both pathogens. In total, 16 time series were analysed including 3947 audits from 474 Spanish farms associated with 302 companies between 2016 and 2019. The monthly time series of the EP index between 2016 and 2019 in Spain (point estimate for Spain was -0.088 with an associated p = 0.073) and different Spanish subregions showed decreasing trend patterns (point estimates for Aragon was -0.028 with an associated p = 0.000 and for Catalonia was -0.064 with an associated p = 0.092), whereas the monthly time series of the CP index increased (point estimate for Spain was 0.004 with an associated p = 0.045 and for Aragon was 0.007 with an associated p = 0.000) over the same period. Additionally, the predictive performance of the estimated models was evaluated at quasi-real time using the data between 2020 and 2021. Results from this evaluation showed that overall, the selected models predicted the evolution of both the EP and CP indices in a reasonable manner being between 90 % prediction intervals. Therefore, time series models constructed in this work could be used to prevent and shorten the response time in implementing of control strategies against these respiratory pathogens minimizing their economic impact associated.

Cryptosporidium spp. are protozoan parasites that cause cryptosporidiosis, an enteric disease that can affect a wide range of vertebrate hosts. Pigs play a potential role in the transmission of Cryptosporidium spp. to humans, although infections are most often subclinical. This study aimed to assess the occurrence and molecular characterization of Cryptosporidium spp. in swine farms located in Aragón, northeastern Spain. Fecal samples (n = 72) were collected from 10 breeding farms, encompassing various production stages (lactation, nursery, fattening, and/or wean-to-finish). Data regarding the type of production system (two- or three-stage), production stages, and farming facilities (the type of flooring and water source) associated with the parasite presence were also analyzed using a questionnaire. The results showed that Cryptosporidium spp. were more frequently detected in fecal samples originating from three-stage production systems (21.9%) compared to two-stage systems (12.5%). Samples from the fattening stage exhibited the highest positivity rate and estimated oocyst count (3.0 oocyst/microscopic field). Furthermore, the molecular characterization of Cryptosporidium spp. revealed the circulation of multiple species both among farms and within the same pig production flow, with Cryptosporidium scrofarum being the most prevalent species (7/72; 9.7%), followed by Cryptosporidium suis (1/72; 1.4%). These findings underscore the importance of the surveillance and molecular characterization of Cryptosporidium spp. for controlling infections in pigs, considering the potential for the zoonotic transmission of this parasite to humans.

One of the main challenges for the sustainability and productivity of the Spanish swine industry is health instability, resulting in significant economic losses. Information on the main swine diseases which affect the Spanish pig industry could help in optimizing the efforts within control programs. This study determined the frequency of occurrence of the main diseases in Spain and the main control tool used, based on perceptions from veterinarians and consultants in a specific survey. Results showed that Streptococcus (S.) suis, E. coli, and coccidia are the most frequent pathogens in the gestation and lactation phase, whereas the most important were Porcine Reproductive and Respiratory Syndrome virus (PRRSV). In the nursery phase, the most frequent were S. suis, E. coli, and PRRSV, the latter being the most important for the participants. Finally, in the fattening phase, PRRSV and Actinobacillus pleuropneumoniae were the most frequent and important pathogen, respectively. Statistical differences among responses were detected with respect to the location and the gestation and lactation phases by farm size. Regarding the tools used for controlling the diseases, vaccination was the main strategy in all production phases, except in the fattening period, in which antibiotherapy was the most common response from the participants. Finally, the improvement of management practices was the most proposed tool, suggesting its importance within control programs.

Influenza D virus (IDV) and co-circulating respiratory pathogens were investigated using RT-qPCR in 316 samples collected from 210 Spanish cattle farms with bovine respiratory disease (BRD) outbreaks between July 2023 and September 2024. Thirty-eight IDV-positive samples, from 30 farms across 15 provinces throughout Spain, were identified. IDV was significantly more frequent in bronchoalveolar lavage samples (22.1%) and nasal swabs (13.5%) compared to lung tissues (5%) and other sample mixtures (5%). All IDV-positive specimens exhibited co-infections, with most (94.7%) harboring three to seven viral and/or bacterial pathogens, highlighting the complexity of BRD outbreaks. Cramer’s V analysis revealed moderate IDV association with Mycoplasma bovis (V = 0.255) and Pasteurella multocida (V = 0.223), and a weaker IDV-bovine coronavirus association (V = 0.202), while IDV association with Histophilus somni (V = 0.025) and bovine herpesvirus 1 (V = 0.000) was negligible. Partial sequences of the hemagglutinin-esterase (HEF) gene from a subset of 13 IDV-positive samples (Ct < 32) were obtained. This confirmed the presence of the two major genetic lineages detected among cattle in Europe, D/OK and D/660, with D/660 exhibiting higher genetic diversity, as determined by DNAsp 6.12 software. This is the first report of IDV infection in Spanish cattle, confirming the circulation of the D/OK and D/660 lineages within the cattle population.

Cryptosporidium spp. oocysts are highly resistant to conventional disinfection methods and have been associated with foodborne outbreaks linked to unpasteurized fruit and vegetable juices. This study aimed to evaluate the effectiveness of Pulsed Electric Fields (PEF) in permeabilizing Cryptosporidium oocysts in water, apple juice, and carrot juice. Oocysts were exposed to monopolar square-wave pulses (3 µs) at electric field strengths ranging from 15 to 35 kV/cm, with treatment times up to 180 µs, and application temperatures between 25 °C and 60 °C. Membrane permeabilization was assessed using propidium iodide uptake via fluorescence microscopy and flow cytometry. Results showed that oocyst permeabilization increased with electric field strength, treatment time, and temperature, with up to 90% permeabilization achieved at 35 kV/cm and 45 °C. Carrot juice treatments yielded higher permeabilization levels than apple juice, attributed to greater electrical conductivity and energy input. Temperatures below 60 °C alone had negligible effects, but synergistically enhanced PEF efficacy. These findings demonstrate that PEF, particularly when combined with mild heat, is a promising non-thermal technology for reducing Cryptosporidium viability in beverages, offering an effective alternative for improving the microbiological safety of minimally processed juices while preserving sensory and nutritional quality.

Background/Objectives: Maternally derived antibody (MDA) levels of porcine circovirus 2 (PCV2) may eventually interfere with humoral response and vaccination efficacy. This study aimed to evaluate the efficacy of a ready-to-use PCV2d and Mycoplasma hyopneumoniae combined vaccine in piglets with different PCV2 MDA levels at vaccination in an experimental inoculation with a heterologous viral genotype. Methods: Forty-eight piglets were allocated into vaccinated (V) and non-vaccinated (NV) groups with high (H) and low (L) PCV2 MDA subgroups (H-V, H-NV, L-V, L-NV). At 3 weeks of age, the piglets received either one dose of vaccine or placebo. Five weeks later, all animals were intranasally challenged with a PCV2b inoculum. Body weight was registered at different time points. Blood samples, peripheral blood mononuclear cells and tracheobronchial lymph nodes (TBLN) were collected and used to assess viraemia, viral load, humoral and cellular responses and histological lesions. Results: The V group showed higher PCV2 antibody levels from challenge onwards, along with a lower percentage of viraemic pigs and reduced viral load in serum at 2 and 3 weeks post-challenge (wpc) and in TBLN tissues compared to the NV group. The H-V group had the highest antibody levels post-challenge, showed no detectable viraemia and had a lower overall amount of virus in tissues. The NV group (especially H-NV) exhibited increased levels of IFN-γ, IFN-α and TNF-α post-challenge. Conclusions: The tested vaccine elicited humoral and cellular immune responses and reduced viral presence in serum and tissues, demonstrating efficacy in a PCV2 subclinical infection model despite high MDA levels at the time of vaccination. Understanding both humoral and cellular immune responses according to different MDA levels can help design more effective vaccination strategies against PCV2.