Search Results (1,812)

Background/Objectives: African swine fever (ASF) is a highly lethal disease of domestic pigs and wild suids that continues to cause substantial economic losses worldwide. Despite recent progress in live attenuated ASF vaccine development, evidence supporting durable protection under repeated exposure conditions representative of endemic settings remains limited. Here, we assessed the long-term safety and protective efficacy of a live attenuated ASFV-G-ΔI177L/ΔLVR vaccine using a repeated-challenge experimental design intended to model re-exposure in ASF-endemic regions. Methods: Vaccinated pigs were subjected to homologous virulent ASF virus challenges at multiple intervals, including repeated challenges (three sequential inoculations) and single challenges administered at 8 and 12 weeks post-vaccination. Results: Across all challenge regimens, vaccinated animals survived and remained clinically healthy, including those receiving three challenges, supporting sustained protection under repeated exposure pressure. Animals challenged at 8 or 12 weeks post-vaccination likewise exhibited complete survival, indicating maintained efficacy through at least 12 weeks. No vaccine-associated adverse clinical outcomes were detected over the study period, and post-challenge viral shedding was minimal. Conclusions: Overall, these data demonstrate that the candidate live attenuated ASF vaccine provides excellent protective efficacy and confers sustained protection against homologous ASF virus infection. This result is expected to be equally applicable under repeated exposure conditions in regions with unstable ASF biosecurity, making it a sufficiently promising model experiment for field application in ASF epidemic areas. However, this is still a vaccine variant, and further studies are planned to evaluate its genomic stability and transmissibility. Full article

Porcine circovirus type 5 (PCV5) is a recently reported porcine-associated CRESS DNA virus, but information regarding its occurrence, genomic characteristics, and evolutionary relationship remains limited. In this study, a total of 100 diagnostic samples collected from clinically diseased pigs from 27 commercial swine farms in 16 cities across seven provinces of China during 2025 were screened for PCV5 using quantitative PCR. PCV5 was detected in 22% (22/100) of the tested samples, with positive samples identified in Henan and Fujian provinces among the sampled regions. PCV5-positive samples were mainly fecal samples and were obtained from pigs showing diarrhea, respiratory signs, wasting, or systemic disease. Co-detection analysis showed that most PCV5-positive samples were also positive for other swine viral pathogens, particularly PCV3, PCV2, and PEDV, indicating that the clinical significance of PCV5 should be interpreted cautiously. Complete genome amplification and sequencing yielded two identical PCV5 genomes from Henan and Fujian provinces. A representative strain, designated PCV5-Henan2025-ZJQ01, was further characterized and deposited in GenBank under accession number PZ496079. The complete genome was 2903 nt in length and contained a positive-sense ORF encoding Cap and a negative-sense ORF encoding Rep, showing a distinct genomic organization compared with classical porcine circoviruses. Phylogenetic analysis based on Rep and Cap amino acid sequences showed that PCV5-Henan2025-ZJQ01 was closely related to previously reported PCV5-related sequences but distinct from classical PCV1–PCV4. These findings provide additional molecular and genomic evidence for PCV5 in Chinese swine diagnostic samples and support the need for continued surveillance and further studies on its epidemiological and pathogenic significance. Full article

This review analyses and logically structures modern intelligent sensor technologies in the context of animal husbandry, feed production, and veterinary medicine. The main research discussed in the article focuses on machine learning based on modern neural network models, computer vision, and sensor systems that are transforming the methods for assessing the health, behavior, and nutrition of farm animals. The first part examines modern approaches to quality control and optimization of mineral and vitamin premixes, including visual inspection using visual sensors and neural networks. Key roles are played by precise dosing, component stability (minerals, vitamins), and the transition to more bioefficient organic forms of micronutrients to reduce environmental impact. Improvements in feed and premix production are analyzed, including automation, energy management, and the use of machine learning for non-destructive quality control, defect detection, mixing homogeneity assessment, and vitamin stability prediction. The second part analyzes methods for animal location and behavior detection. This article presents computer vision-based systems, including modifications of YOLO, for automatically tracking and classifying key behavioral patterns (lying down, standing, feeding, and aggression) in cattle and pigs, even in crowded conditions. It also discusses the use of ultra-wideband (UWB) systems and accelerometers combined with machine learning for high-precision positioning and detection of specific behavioral anomalies, such as lameness and playfulness. The third section focuses on the application of machine learning in veterinary diagnostics, including the automated interpretation of medical images (X-ray, ultrasound, and MRI) as sensor data streams for the diagnosis of cardiovascular, oncological, and orthopedic diseases in farm and small animals. Furthermore, the article examines the use of machine learning models for proactive disease diagnosis in farm animals and poultry based on multimodal data and image analysis. Considerable attention is given to methods and tools for radiometric diagnosis of animal diseases at an early stage using microwave sensors, as well as laser therapy and surgery in veterinary medicine. The review concludes that the integration of intelligent systems enables a transition to data-driven livestock management, significantly improving animal welfare and, consequently, the efficiency and sustainability of agricultural production. Full article

Weaning stress and immune challenges can negatively affect the health and performance of young pigs by inducing inflammatory responses. Functional protein sources, such as enzymatically hydrolyzed whole blood (EHB), may help alleviate inflammation and improve immune status during stressful conditions. A total of 20 late-weaned pigs [Landrace × Yorkshire × Duroc], 42 days of age, with an initial body weight of 15.34 ± 1.17 kg, were used in a 2-week experiment. Pigs were allotted to a 2 × 2 factorial arrangement with two dietary protein sources [plasma protein (PP) and/or EHB] and two immune challenges (saline or LPS). The four experimental groups were as follows: (1) Plasma-Sal, PP diet + saline injection; (2) Plasma-LPS, PP diet + LPS injection (100 μg/kg BW); (3) EHB-Sal, EHB diet + saline injection; and (4) EHB-LPS, EHB diet + LPS injection (100 μg/kg BW). Each treatment consisted of five replicate pens with one pig/pen. Pigs fed either protein diet with and without LPS showed no (p > 0.05) difference in their nutrient digestibility and microbial population. However, pigs challenged with LPS exhibited a higher (p < 0.05) rectal temperature, with significant differences observed at 6 h and 12 h post-injection (p < 0.001). Dietary effects (p < 0.05) were observed for IL-6 and TNF-α concentrations, with pigs fed EHB exhibiting lower values compared with those fed the PP diet following LPS challenge. Consistent with an inflammatory response, pigs challenged with LPS showed elevated (p < 0.05) IL-6 and TNF-α concentrations, together with increased white blood cell and lymphocyte counts, at 12 h post-challenge. Moreover, significant (p < 0.05) diet × LPS interactions were detected for IL-6 and TNF-α concentrations at 6 h post-challenge, indicating that dietary EHB attenuated the inflammatory response induced by LPS. In summary, a diet formulated with EHB showed a reduced effect of LPS challenge in pigs, making it promising as a functional dietary protein source for improving immune resilience in weaned piglets. Full article

Accurate pig counting during herd transfers is fundamental to effective livestock management in large-scale swine production, yet existing methods struggle with bidirectional passages, boundary oscillations, and occlusion in real corridor environments. This study proposes an integrated system combining an improved YOLO-based detection model with a Hysteresis-based Multi-frame Temporal Confirmation Counting Strategy (HMTC). The YOLO11s baseline was enhanced using lightweight RepViT blocks, dynamic upsampling (DySample), and shape-aware bounding box regression (Shape-IoU). The resulting model achieves a mAP₅₀ of 0.982 with a compact architecture of 8.28M parameters, representing a 12.3% reduction relative to the baseline while improving detection accuracy. To address bidirectional counting challenges, the HMTC strategy utilizes hysteresis-based region classification, temporal confirmation, and trajectory verification to suppress boundary jitter and ensure directional correctness. Evaluated on nine videos from a single transfer corridor, the proposed system achieves an overall counting accuracy of 99.21% on this test set and runs in real time on an embedded edge device at over 30 FPS without loss of counting accuracy. Together, the improved detection model and HMTC counting strategy provide a cohesive approach to pig passage counting, validated here under a single transfer-corridor condition; these results offer a promising basis for automated animal inventory management, pending further validation across more diverse farm environments. Full article

African swine fever in both domestic and wild pig populations is caused by the extremely infectious African swine fever virus (ASFV). It seriously endangers biodiversity and results in large financial losses for the worldwide pork sector. The major capsid protein p72 is molecularly chaperoned by the ASFV pB602L protein, which is essential to viral assembly. Furthermore, as a nonstructural protein expressed at late stages of infection, pB602L induces a distinct antibody response that may complement existing serological assays based on structural proteins. Given its strong immunogenicity, pB602L represents a promising antigen for developing supplementary diagnostic tools for African swine fever (ASF). In this study, we successfully generated and separated the ASFV pB602L protein, and we verified its responsiveness using serum from pigs infected with ASFV. Additionally, we produced four monoclonal antibody-specific hybridoma cell lines that targeted the pB602L protein exclusively. These cell lines demonstrated high immunoreactivity and responsiveness toward ASFV pB602L. These results highlight the potential enhancement of diagnostic skills. We have detected two previously unknown linear B-cell epitopes (¹³⁸TIDSFL¹⁴³ and ¹⁶⁴TNVDTC¹⁶⁹) using overlapping peptide and truncated protein fragment analysis. Due to their high degree of conservation across various ASFV strains, these epitopes offer trustworthy candidates for the creation of particular diagnostic instruments. This study expands the known ASFV antigenic repertoire by systematically mapping immunodominant epitopes of pB602L. The identified epitopes provide potential molecular targets for the rational design of multi-epitope subunit vaccines. Full article

One of the keys to improving feed conversion rates in Precision Livestock Farming (PLF) is the early identification of growth impediments. However, the swine farming data collected by Electronic Feeding Station (EFS) are often disorganized and lack effective labeling. Data from healthy pigs are frequently intermixed with that from sick pigs, leading to label leakage and survivor bias in models, particularly when age is included as a feature. To address these known issues, this study breaks away from traditional modeling methods. First, we clean and classify the time-series data from electronic feeding stations, using age-cohort baselines as one of the criteria for determining high and low productivity, thereby avoiding problems such as label leakage. Next, we construct a high-dimensional feature matrix that captures dynamic derivatives such as feeding acceleration and weight gain acceleration, which together serve as behavioral feature fingerprints. To test the system, we optimized the mixed-model algorithm and evaluated the model based on behavioral deviations among individual pigs after removing all absolute age labels. Our results indicate that the full-feature model achieved an ROC-AUC of 0.778 and an F1-score of 0.4137 at the optimal threshold. Interestingly, SHAP attribution analysis revealed that “intake peer deviation,” “Cumulative Intake and Lifetime Avg Intake,” and “feeding acceleration” served as precursors to low productivity and growth retardation in this dataset, with these factors proving more significant than absolute feed intake or age. Our ablation experiments confirmed that a model based solely on behavioral features (excluding age labels) maintained an ROC-AUC of 0.773, successfully decoupling pig growth performance from growth stage. Our model can detect changes in feeding dynamic signatures at an average of 12.3 days, thereby providing insights for pig growth assessment, health monitoring, or more informed culling decisions. Full article

African swine fever (ASF) is a highly fatal, febrile infectious disease of domestic pigs and wild boars caused by the African swine fever virus (ASFV). Recently, highly virulent recombinant ASFVs with chimeric genomes derived from p72 genotype I and II viruses have emerged in China, Vietnam, and Russia. These genotype I/II recombinants can evade immunity induced by genotype II–based vaccines, thereby complicating disease control efforts. To address this challenge, a novel duplex quantitative PCR (qPCR) assay was developed to simultaneously detect and differentiate genotypes I, II, and I/II recombinants in a single reaction. The assay exhibited high sensitivity and specificity, with a reliable detection limit of 10 copies/reaction for genotype I and II ASFV DNA. Validation using clinical samples collected in northern Vietnam in 2025 confirmed a robust performance in accurately distinguishing circulating genotype II viruses from recombinant genotype I/II viruses, including the detection of potential co-infection. Whole-genome sequencing of selected positive samples further corroborated these findings. Overall, this qPCR assay provides a precise and efficient tool for identifying currently circulating ASFV genotypes, thereby facilitating improved disease surveillance and supporting a comprehensive understanding of the evolving epidemiological landscape of ASF in regions with increasing viral genetic diversity. Full article

Rapid and reliable detection methods are essential for routine monitoring of environmental hygiene on farms. This pilot study evaluated luminometers (LUM) and mobile flow cytometer (MFC) for assessment of surface organic contamination in farrowing units. The study was conducted on two pig farms after animal removal prior to sanitation, with sampling performed at heated pads, pen walls, and corridors. ATP measurements were carried out using three luminometers (Clean-Trace™ LM1, EnSure, and SystemSURE Plus), and residual particles were detected using a mobile flow cytometer (Cytoquant). Microbiological cultivation (TMC 36 °C) was additionally included. Significant differences in log RLU values were observed between LUM, with large effect sizes indicating a substantial influence of device type on RLU values. A high correlation was confirmed only between EnSure and SystemSURE Plus (r_s = 0.81–1.00; p < 0.05), and no relationship was confirmed between LUM and MFC (r_s = −0.49–0.77; p > 0.05). Correlations between rapid detection methods and microbiological cultivation were inconsistent. Corridors demonstrated the highest microbiological contamination, whereas MFC identified heated pads as sites with increased residual particulate contamination. The results indicate that LUM, MFC, and microbiological cultivation characterize different dimensions of environmental contamination and should therefore be interpreted as complementary rather than interchangeable methods. Full article

One-lung ventilation (OLV) is performed during lung surgeries by ventilating a single lung, while collapsing the operative lung to provide surgical exposure within the thoracic cavity. While a lung-protective ventilation strategy is recommended during OLV, increasing the fraction of inspired oxygen (FiO₂) or tidal volume (V_T) may be required to prevent hypoxemia during surgery. Unfortunately, these increases are associated with postoperative lung injury. Using a porcine model of OLV, our project aims to determine if high FiO₂ or V_T during OLV contributes to elevation of pro-inflammatory lipid mediators postoperatively. Fifteen three-month-old farm-bred pigs underwent left upper lobectomy requiring OLV. Pigs were exposed to one of three ventilation parameters: normoxic low V_T lung-protective ventilation LPV-NO, n = 5, FiO₂ < 50%, V_T = 6 mL/kg), hyperoxic lung-protective ventilation (LPV-HO, n = 5, FiO₂ >100%, V_T = 6 mL/kg), or normoxic high V_T (injurious mechanical ventilation) (IMV, n = 5, FiO₂ < 50%, V_T = 10–12 mL/kg). Arterial plasma was collected before and after OLV, and lipids were detected via LC-MS-MS. Lipidomic analysis demonstrated a statistically significant increase (FC = 2, p ≤ 0.05) in lysophosphatidylethanolamines (LPE 18:3, LPE 20:4, LPE 18:2, LPE 22:6), free fatty acids (FFA 20:4), phosphatidylserine (PS 38:5), lysophosphatidylcholine (LPC 18:1, LPC 18:3, LPC 22:6), triglyceride (TG 18:2-18:2-20:4), free fatty acids (FFA 20:5), linoleyl-carnitine molecules (C18-2 Linoleoyl Carnitine), and phosphatidylethanolamines (PE 36:5) in LPV-HO. IMV resulted in a significant increase (FC = 2, p ≤ 0.05) in triglyceride (TG 18:2-18:2-20:4), diglyceride (DG 18:1-20:4, DG 16:0-20:4), linoleyl-carnitine molecules (C18-2 Linoleoyl Carnitine), and free fatty acids (FFA 20:5). There was no significant change in lipid biomarker levels following LPV-NO post-surgery. Our lipidomic analysis supports that both high FiO₂ and V_T contribute to systemic lipid metabolic derangements. Lipids that were elevated in LPV-HO and IMV are associated with multiple inflammatory pathways implicated in lung injury. This suggests that intra-operative anti-inflammatory therapies targeted to these lipid pathways may reduce or prevent postoperative pulmonary complications after lung surgery. Full article

Bovine viral diarrhea (BVD) is a globally significant disease that adversely affects cattle health and productivity, including in India. It is caused by three bovine pestiviruses: bovine viral diarrhea virus 1 (BVDV-1), BVDV-2, and HoBi-like pestivirus (HoBiPeV), which belong to the Pestivirus genus within the Flaviviridae family. Despite the prevalence of all three pestivirus species in India, no commercial vaccine based on the local circulating strain is currently available. This study evaluates the safety, immunogenicity, and protective efficacy of an inactivated whole-virus BVD vaccine, based on an Indian BVDV-1 strain. The virus was propagated in MDBK cells, inactivated using 3 mM binary ethylenimine (BEI) for 24 h at 37 °C, and formulated with Montanide ISA 61 VG (SEPPIC) in a 50:50 water-in-oil emulsion. Vaccine safety was confirmed in both guinea pigs and bovine calves, with no adverse effects observed. Immunogenicity testing in guinea pigs (n = 6) showed neutralizing antibody titres up to 9 log₂ (1/512). In calves aged 9–12 months (n = 3), the vaccine elicited strong humoral and cell-mediated immune responses, with mean neutralizing antibody titres against the homologous BVDV-1 strain reaching 14 log₂ (1/16,384). Neutralizing antibody levels remained detectable for up to 12 months post vaccination with sustained mean titres of 7 log₂ (1/128). Notably, titres reported to be adequate for fetal protection (≥9 log₂ or ≥1/512 were maintained for five months following vaccination. Challenge studies demonstrated complete protection of vaccinated calves against homologous BVDV-1 acute infection. In addition, the vaccine conferred partial cross-protection against heterologous strains including BVDV-2 and HoBiPeV. In a field trial involving 125 cattle, 74% of animals developed protective neutralizing titres (≥7 log₂ or ≥1/128), while 48% achieved titres reported to be adequate for fetal protection (9 log₂ or 1/512). Furthermore, 92% of vaccinated cattle maintained neutralizing antibody titres of at least 6 log₂ (≥1/64) for up to six months post-booster vaccination. A strong positive correlation was observed between guinea pig and bovine antibody responses (R² = 0.6809; p < 0.0001), indicating the potential of guinea pigs as a predictive model. Vaccine stability was confirmed for up to 8 months when stored at 4 °C, as demonstrated by the immunogenicity in guinea pigs. Collectively, these findings demonstrate that the locally developed inactivated BVDV-1 vaccine is safe, highly immunogenic, and capable of providing protective immunity against BVDV-1 infection, supporting its potential use in BVD control programs in India. Full article

Background/Objectives: Post-weaning diarrhea remains a major challenge in pig production worldwide. Enterotoxigenic Escherichia coli (ETEC) encoding fimbriae of the F4 type and producing the heat-stable enterotoxin, STb, are one of the important causes of this disease. The aim of the current study was to evaluate whether vaccination of pregnant sows with a novel capsid virus-like particle (cVLP)-based vaccine against F4 and STb (cVLP-FaeG/cVLP-STb) could enhance performance in piglets born after such vaccinated sows. Methods: A field trial was conducted in a commercial sow-to-finisher pig herd. Thirty-five sows were vaccinated twice with the cVLP-FaeG/cVLP-STb vaccine prior to farrowing, while thirty-five control sows were vaccinated twice with commercial vaccines normally used in the herd. Piglets were followed until eight weeks post-weaning to assess antibody responses, diarrhea and treatment incidences, pathogen shedding, and growth performance. Results: Piglets born from immunized sows receiving the cVLP vaccine showed significantly higher serum antibody levels against ETEC F4 throughout the post-weaning period (p ≤ 0.021). The frequency of pathogen detection was similar between groups, while piglets in the cVLP group exhibited significantly lower diarrhea scores at week 6 (p = 0.047), showed a trend of requiring fewer treatments (p = 0.06) and had significantly higher final body weight (p = 0.048). In addition, the cVLP group showed a significantly greater average daily gain over the study period (p = 0.037). Conclusion: Sow immunization with the cVLP vaccine enhanced passive immune protection of piglets, resulting in reduced antimicrobial treatment 2 weeks post-weaning and improved growth performance. Full article

Background/Objectives: African Swine Fever (ASF) represents one of the most serious threats to animal health and global food security. The causative agent of ASF is the African swine fever virus (ASFV), a DNA virus belonging to the Asfarviridae family. Here, we describe ex vivo results for an original anti-ASFV vaccine approach based on the cellular immune response induced by extracellular vesicles (EVs) engineered to express four ASFV proteins. EV engineering was achieved by expressing a DNA vector encoding a biologically inactive HIV-1 Nef protein (Nef^mut), which exhibits unusually high efficiency of incorporation into EVs, even when fused to foreign proteins. Previous studies have demonstrated that intramuscular injection of Nef^mut-based vectors leads to the engineering of Evs, spontaneously released by muscle cells, and induction of antigen-specific CD8⁺ T cell immunity. Methods: We designed DNA vectors expressing the fusion products between Nef^mut and each of the four ASFV structural proteins p30, p54, pp62, and p72. Engineered EVs were molecularly characterized by Western blot and nanotrack analysis, and their potential immunogenicity was assessed by priming and cross-presentation assays. Results: We assessed that the four fusion proteins were successfully expressed in transfected mammalian cells, with the release of valuable amounts of engineered EVs. When immature swine dendritic cells were challenged with the engineered EVs and then co-cultivated with autologous peripheral blood lymphocytes in priming assays, lymphocyte subpopulations specifically reacting against each ASFV antigen were elicited, as detected by an IFN-γ ELISpot assay. In addition, we provide evidence that the Nef^mut-based fusion products incorporated into the engineered EVs can be cross-presented by professional antigen-presenting cells, leading to cross-priming of autologous lymphocytes. Conclusions: These results represent the best premise to go forward with experiments examining immunogenicity and antiviral efficiency in pigs. Full article

Salmonella enterica serovar Choleraesuis (S. Choleraesuis) is a swine-adapted zoonotic pathogen for which multidrug resistance has become an increasing concern. An antimicrobial resistance profile and multidrug resistance pattern in S. Choleraesuis isolates from diseased pig in Taiwan have been reported. This study was designed as a secondary molecular and statistical analysis of 272 isolates recovered from diseased pigs in Taiwan whose minimum inhibitory concentration phenotypes had been reported previously. Using this isolate collection, we evaluated phenotype–genotype concordance and coresistance patterns by combining antimicrobial susceptibility data with targeted polymerase chain reaction screening and quinolone resistance-determining region sequencing. Multidrug resistance was detected in 97.8% of isolates. Among the screened acquired determinants, blaTEM was the most prevalent gene (94.5%), followed by floR (55.9%) and mcr-1 (12.9%); mcr-3 was detected in only one isolate. Significant phenotype–genotype concordance was observed for aphA-1 with kanamycin resistance and mcr-1 with colistin resistance. Sequence analysis of 60 enrofloxacin-resistant isolates revealed that high-level resistance was associated with combined gyrA and parC substitutions, with additional gyrB-region polymorphisms observed in isolates showing the highest minimum inhibitory concentrations. These findings demonstrate frequent coresistance patterns and complex resistance mechanisms in this lineage and support continued genomic surveillance and prudent antimicrobial stewardship in the swine sector. Full article

Respiratory disease remains a major challenge in pig production. This two-room pilot study evaluated whether room-level acoustic monitoring combined with physiological measurements could provide an early warning after an experimental Klebsiella pneumoniae challenge. Forty growing pigs balanced by sex and body weight were housed for 28 days in one control room and one challenged room (20 pigs/room; four pens/room). Challenged pigs were intranasally inoculated on days 8, 12, 16, and 20 with a culture whose dose was retrospectively verified by serial-dilution plating. Nasal and fecal samples were cultured on Klebsiella ChromoSelect agar, and colonies with expected morphology were enumerated as presumptive Klebsiella/K. pneumoniae colonies. A fine-tuned Audio Spectrogram Transformer (AST) classified five sound classes from facility-specific audio and was evaluated by group-blocked hold-out testing, five-fold group-blocked cross-validation, temporal deployment validation, and window-threshold sensitivity analysis. The model achieved hold-out macro-F1 of 0.947, five-fold macro-F1 of 0.928 ± 0.019, and 24 h deployment macro-F1 of 0.914. Presumptive nasal bacterial load was higher in challenged pigs at 1-week post-inoculation (log₁₀ 4.03 vs. 0.67). Group-size-standardized cough detections were also higher in the challenged room (54.84 vs. 36.80 detections/day), and daily coughing first exceeded the baseline threshold on day 8. Thresholds of 0.764 (control) and 1.115 (treatment) were obtained from an integrated score that included coughing, sneezing, ear temperatures, rectal temperature, and respiration rate; the treatment score and treatment–control contrast score first surpassed the threshold on day 8, and daily multimodal scores varied between groups (t = −6.636, p < 0.001). Integrated score improved discrimination of post-inoculation disturbance compared with cough detections alone (leave-one-day-out AUROC: 0.94 vs. 0.88). Because each condition was represented by one room, findings are exploratory temporal contrasts, not replicated treatment effects or a stand-alone diagnostic test. Full article