Application of hot water and cold air to reduce bacterial contamination on broiler carcasses.

Two physical treatments (heat via water bath and cold air) with various temperatures (20/70/75/80°C and - 80/-90°C) and exposure times (20, 30, 40 s) were carried out to identify a decontaminating effect on zoonotic pathogens on broiler carcasses. Subsequently, carcasses were analyzed for thermotolerant Campylobacter (C.), Salmonella, Escherichia (E.). coli and total colony count (TCC). Moreover, for the hot water treatment, qPCR with viable/dead differentiation (v-qPCR) was applied to detect viable but non-culturable cells (VBNC) of Campylobacter, referred to as intact but putatively infectious units (IPIU). Hot water immersion was tested on carcasses inoculated with C. jejuni and Salmonella, while cold air treatment was evaluated for naturally contaminated carcasses of broiler flocks colonized with Campylobacter. For hot water treatment, the statistically significant reducing effect was about 1 log10 CFU/ml for both Salmonella and Campylobacter for 70-80°C and 20/30 s treatments. The effect of heat treatment for Campylobacter was smaller when samples were analyzed with v-qPCR with reductions of 0.5-0.8 log10 IPIU/ml in mean. Cold air treatments at -90°C were effective in reducing the mean contamination level of Campylobacter by 0.4-0.5 log10 CFU/ml at all exposure times (p < 0.05). Hot water treatments showed a decreasing trend on TCC by 0.6-0.9 log10 CFU/ml (p < 0.05). TCC counts were not significantly affected by cold air treatment. For E. coli no statistically significant reductions were observed by hot water treatment. The cold air treatment at -90°C for 20 and 40 s led to a reduction of E. coli by 0.4 and 0.8 log10 CFU/ml (p < 0.05), respectively. Treatment of carcasses with higher bacterial levels tended to show higher reduction. The research demonstrated that the efficacy of physical treatments for decontamination of broiler carcasses was more pronounced for hot water immersion than for cold air exposure. In conclusion, the results shed light on the potential application of these physical treatments in practice to reduce the quantitative load of contaminating pathogens to enhance food safety in the broiler meat production.

Influences on the Decision to Euthanize a Compromised Pig.

The decision to euthanize a compromised pig can be challenging for pig farmers and veterinarians. To understand more about the challenges in Germany, a cross-sectional online survey was conducted. Based on a hybrid design, the responses of 39 veterinarians and 62 pig farmers were analyzed to generate a list of common clinical signs associated with the euthanasia of sows, fatteners, and piglets. Moreover, a set of influences on the farm, due to economic and personal considerations, were found to shape the decision-making process. The two most salient reasons outlined for the delay of timely euthanasia were uncertainty and misinterpretation of the chance for healing. The lack of valid clinical signs or a sound justification was most frequently mentioned as a challenge to the general decision-making process. In summation, this study highlights the need to generate a valid taxonomy for clinical signs that includes their development in a compromised pig over time. Future studies should elaborate on the justification of euthanasia decisions to facilitate the resolution of ethical dilemmas among the involved pig farmers and veterinarians. Lastly, the results suggest that clinical reasoning and consultation skills should be included when decision-making behavior is to be trained.

Antimicrobial Resistance Surveillance: Data Harmonisation and Data Selection within Secondary Data Use.

Resistance to last-resort antibiotics is a global threat to public health. Therefore, surveillance and monitoring systems for antimicrobial resistance should be established on a national and international scale. For the development of a One Health surveillance system, we collected exemplary data on carbapenem and colistin-resistant bacterial isolates from human, animal, food, and environmental sources. We pooled secondary data from routine screenings, hospital outbreak investigations, and studies on antimicrobial resistance. For a joint One Health evaluation, this study incorporates epidemiological metadata with phenotypic resistance information and molecular data on the isolate level. To harmonise the heterogeneous original information for the intended use, we developed a generic strategy. By defining and categorising variables, followed by plausibility checks, we created a catalogue for prospective data collections and applied it to our dataset, enabling us to perform preliminary descriptive statistical analyses. This study shows the complexity of data management using heterogeneous secondary data pools and gives an insight into the early stages of the development of an AMR surveillance programme using secondary data.

In-depth analysis of the medical supply for indigenous people in North-Eastern Colombia: a dominance of infectious diseases and only insufficient therapeutic options.

BACKGROUND: Colombian indigenous Wiwas are exposed to a variety of partly complex medical conditions with a predominance of infectious diseases. The study provided here aims at verifying of falsifying previous suspicions on therapeutic shortcomings and neglect of disease categories. MATERIAL AND METHODS: Local diagnoses within various subpopulations of indigenous Wiwas obtained by a study physician and local health brigades and health points between 2017 and 2018 were coded following the ICD 10 classification from 2019. Proportions of diagnoses per ICD-10 sub-chapter were evaluated to find diseases and to rank the occurrence of diagnoses in the population of indigenous people. Thereafter, the available medication provided by the indigenous health care provider Dusakawi for the treatment of the indigenous patients was analyzed in regard of its sufficiency to cover the recorded diseases. RESULTS: The majority of the diseases found in the communities cannot at all (32%) or only partially (56%) be treated according to available guidelines. Only few (12%), predominantly infectious diseases, were covered completely by the provided medication. Notably, there are some ICD chapters with diseases that do only rarely appear at all in the gained datasets, e.g., complications during birth, mental disorders or cancer. CONCLUSIONS: An expansion and revision of the medical supply for the indigenous population of the Sierra Nevada de Santa Marta is needed. An emergency kit for medical brigades and health points should be provided and in place. Awareness for neglected diseases needs to be created.

Foot lesions and forelimb skin abrasions in suckling piglets: development and risk factors.

BACKGROUND: Foot lesions in suckling piglets have been associated with poor flooring in several studies and were recently proposed to be indicative of swine inflammatory and necrosis syndrome. However, identical findings are also the typical outcome of various non-infectious causes; thus, further risk analysis is needed. The objective of this study was to describe the development of heel bruising, coronary band lesions and forelimb skin abrasion in suckling pigs up to 5 days of age. Furthermore, the effects of various intrinsic and extrinsic factors were examined. On each of four commercial piglet-producing farms, piglets from two or three batches of eight sows were studied. The piglets were included within 18 h after birth. Each piglet was individually scored four times. The score for the heels differentiated six (0-5) and for the coronary band and forelimb skin abrasion three stages (0-2). The body weight was measured two times. The effect of the floor was estimated by allocating the sows randomly to farrowing pens equipped with either soft rubber mats covered with litter or fully slatted plastic floors. RESULTS: The final analysis comprised data from 1045 piglets. Foot lesions were not found at birth but started to develop on day 1. On day 5, heel bruising was found in 94%, main claw coronary band lesions in 49% and forelimb skin abrasion in 73% of the piglets. In a multifactorial logistic regression analysis, it was shown that a slatted plastic floor significantly increased the odds of heel bruising and coronary band lesions, while a rubber floor with litter increased the odds of forelimb skin abrasions. CONCLUSION: Foot and forelimb lesions in new-born piglets are mainly induced by the floor. The effect of slatted plastic floors on heel bruising showed an overwhelming OR of 52.89 (CI 26.29-106.43). Notably, coronary band lesions in young suckling piglets occur on slatted as well as non-slatted floors, indicating that the piglets incur these injuries not only from the wedging of their feet into the gaps between slats but also from contact with the floor while suckling. Based on these findings, preventive measures should be redirected to the improvement of the floor in the farrowing pen, particularly in the area under the sow's udder.

Agar Contact Method as a Valuable Tool to Identify Slaughter Hygiene Deficiencies along the Slaughter Process by Longitudinally Sampling Pig Skin Surfaces.

Examinations of total viable counts (TVCs) and Salmonella spp. on the skin of individual pigs during the slaughter process are useful to identify abattoir-specific risk factors for (cross-)contamination. At seven process stages (lairage to before chilling), pigs were bacteriologically investigated by repeatedly sampling the same animals using the agar contact method. The mean TVC of all pigs increased significantly at the first three tested process stages (mean count, after delivery: 5.70 log cfu/cm2, after showering: 6.27 log cfu/cm2, after stunning: 6.48 log cfu/cm2). Significant mean TVC reductions occurred after scalding/dehairing (mean count: 3.71 log cfu/cm2), after singeing/flaming (2.70 log cfu/cm2), and after evisceration (2.44 log cfu/cm2) compared with the respective preceding process stages. At the end of the slaughter line and before chilling, the mean TVC was 2.33 log cfu/cm2, showing that the slaughter process reduced contamination significantly. The slaughter process effectively reduced even very high levels of incoming TVCs, since at the individual animal level, at the end of the slaughter process, there was no difference in the TVCs of animals with initially high and initially low TVCs. Additionally, 12 Salmonella spp. isolates were recovered from 12 different pigs, but only until the stage after scalding/dehairing. Overall, the agar contact method used is valuable for detecting hygiene deficiencies at slaughter, and is animal-equitable, practical, and suitable for use on live animals.

Chagas Disease: Medical and ECG Related Findings in an Indigenous Population in Colombia.

Chagas Disease (CD) is highly prevalent among the indigenous populations in the Sierra Nevada de Santa Marta, Colombia. Villages examined show prevalence rates ranging from 43.6% up to 67.4%. In the present study, associated medical conditions were assessed with a particular focus on ECG alterations. CD diagnosis was based on a rapid test, two different ELISAs, and a specific and highly sensitive Chagas real-time PCR. In both CD positive and CD negative patients, relations of the status and medical (physical examination-based, questionnaire-based) and/or electrocardiogram-based findings were investigated. As expected, CD-associated symptoms and complaints were predominantly found in CD-positive patients. Interestingly, ECG-findings were found to show the potential of leading to early CD diagnosis because ECG alterations were already seen in early stagechanges of the disease. In conclusion, although the observed ECG changes are unspecific, they should be considered as an indicator for a CD screening and, in case of positive results, an associated early treatment of the disease.

[Therapy-relevant antibiotic resistances in a One Health context]. / Therapierelevante Antibiotikaresistenzen im One-Health-Kontext.

One Health refers to a concept that links human, animal, and environmental health. In Germany, there is extensive data on antibiotic resistance (AMR) and multidrug-resistant (micro)organisms (MDRO) in human and veterinary medicine, as well as from studies in various environmental compartments (soil, water, wastewater). All these activities are conducted according to different specifications and standards, which makes it difficult to compare data. A focus on AMR and MDRO of human therapeutic importance is helpful to provide some guidance. Most data are available across sectors on methicillin-resistant Staphylococcus aureus (MRSA) and multiresistant Enterobacterales such as Escherichia coli and Klebsiella pneumoniae. Here, the trends of resistance are heterogeneous. Antibiotic use leads to MRE selection, which is well documented. Success in minimizing antibiotic use has also been demonstrated in recent years in several sectors and could be correlated with success in containing AMR and MDRO (e.g., decrease in MRSA in human medicine). Sector-specific measures to reduce the burden of MDRO and AMR are also necessary, as not all resistance problems are linked to other sectors. Carbapenem resistance is still rare, but most apparent in human pathogens. Colistin resistance occurs in different sectors but shows different mechanisms in each. Resistance to antibiotics of last resort such as linezolid is rare in Germany, but shows a specific One Health correlation. Efforts to harmonize methods, for example in the field of antimicrobial susceptibility testing and genome-based pathogen and AMR surveillance, are an important first step towards a better comparability of the different data collections.

Canine olfactory detection of SARS-CoV-2-infected humans-a systematic review.

PURPOSE: To complement conventional testing methods for severe acute respiratory syndrome coronavirus type 2 infections, dogs' olfactory capability for true real-time detection has been investigated worldwide. Diseases produce specific scents in affected individuals via volatile organic compounds. This systematic review evaluates the current evidence for canine olfaction as a reliable coronavirus disease 2019 screening tool. METHODS: Two independent study quality assessment tools were used: the QUADAS-2 tool for the evaluation of laboratory tests' diagnostic accuracy, designed for systematic reviews, and a general evaluation tool for canine detection studies, adapted to medical detection. Various study design, sample, dog, and olfactory training features were considered as potential confounding factors. RESULTS: Twenty-seven studies from 15 countries were evaluated. Respectively, four and six studies had a low risk of bias and high quality: the four QUADAS-2 nonbiased studies resulted in ranges of 81%-97% sensitivity and 91%-100% specificity. The six high-quality studies, according to the general evaluation system, revealed ranges of 82%-97% sensitivity and 83%-100% specificity. The other studies contained high bias risks and applicability and/or quality concerns. CONCLUSIONS: Standardization and certification procedures as used for canine explosives detection are needed for medical detection dogs for the optimal and structured usage of their undoubtful potential.

Case Study: Using a Shared International Database to Document Veterinary Consumption of Antibiotics in Pakistan.

In this paper, we present a case study of Pakistan documenting the use of antimicrobial drugs in poultry flocks in the VetCAb-ID database. Unlike other databases, this system allows international users to upload their data directly. Based on expert interviews and a review of the latest publications on the topic, we provide an alternative approach to harmonizing data collection among countries. This paper will provide impetus to formulate joint requirement documentation for an AMU database on a global level that international users can adapt for their own purposes and projects.

Carriage of Extended Spectrum Beta Lactamase-Producing Escherichia coli: Prevalence and Factors Associated with Fecal Colonization of Dogs from a Pet Clinic in Lower Saxony, Germany.

Extended spectrum beta-lactamase (ESBL)-producing Escherichia coli are an emerging problem in veterinary and human medicine. Our study concentrated on the estimation of the prevalence and factors associated with the carriage of ESBL-producing E. coli in dogs who visited a veterinary clinic in northern Germany in 2017. For this reason, 1000 patients (healthy and sick dogs) were tested, resulting in 1000 samples originating from rectal swabs. Additional data were collected using a self-reported questionnaire that was completed by the dog owner. Factors associated with ESBL carriage were considered for further modeling if p < 0.05 using a two-sided Fisher test. Using a backward elimination procedure, the variables for the final multivariable logistic regression model were identified. In total, 8.9% of the dogs tested were positive for carriage of ESBL-producing E. coli. Seven factors were associated with the colonization of dogs with ESBL-E. coli within the multivariable model, namely husbandry system (p = 0.0019, OR = 3.00; 95% CI: 1.50-6.00), contact with puppies (p = 0.0044, OR = 2.43; 95% CI: 1.32-4.46), feeding of raw meat (p = 0.011, OR = 2.28; 95% CI: 1.21-4.31), food residues (p = 0.0151, OR = 2.31; 95% CI: 1.18-4.53) and food supplements (p = 0.0487, OR = 0.426; 95% CI: 0.18-0.96), and antibiotic treatments of dogs (p = 0.0005, OR = 3.030; 95% CI: 1.62-5.68) or owners (p = 0.041, OR = 2.74; 95% CI: 1.04-7.19) prior to the study. These factors refer to the animals themselves as well as to the owners and their habits or medical treatments. Although the causality and direction of transmission from owners to their dogs cannot be proven, the factor of antibiotic treatment of the owner is clearly associated with the dog's status.

Rate of surgical site and urinary tract infections in dogs after cessation of antibiotics following spinal surgery.

BACKGROUND: Excessive use of antimicrobials and the increasing occurrence of antimicrobial resistance are major challenges in both human and veterinary medicine. The role of prophylactic antimicrobial therapy in orthopaedic and neurosurgeries in dogs can be questioned. The aim of this study was to evaluate the rate of surgical site infections (SSI) and urinary tract infections (UTI) in dogs after cessation of antibiotics following spinal surgery. METHODS: Electronic patient records from January 2018 to December 2019 were retrospectively reviewed to identify dogs that underwent spinal surgery (n = 158). Antimicrobial drug use and the presence of SSI and UTI were recorded. RESULTS: Overall, SSI developed in 1.3% of dogs that underwent spinal surgery, while UTI developed in 8.2%. Multidrug-resistant (MDR) bacteria were detected in 5.1% of dogs. The rates of SSI, UTI and MDR did not differ significantly between dogs that received postoperative antimicrobial therapy and those that did not. LIMITATIONS: The main limitation of this study was its retrospective design. CONCLUSION: Overall, the SSI rate in this study was low. Cessation of postoperative antimicrobial use in dogs following spinal surgery did not have a negative effect on either SSI development or the occurrence of UTI.

Use of Antimicrobials by Class in Pigs in Germany-A Longitudinal Description Considering Different International Categorisation Systems.

Antimicrobial usage in both human and veterinary medicine is considered one of the main drivers of antimicrobial resistance; its reduction poses a serious challenge. To analyse the associations between usage and resistance, data from monitoring systems and classification of all antimicrobial substances are crucial. In this analysis, we investigated longitudinal data collected between 2013 and 2020 within the Veterinary Consumption of Antibiotics project from pig farms in Germany, including all antimicrobial classes, but focusing on critically important antimicrobials: third- and fourth-generation cephalosporins, fluoroquinolones, macrolides, and polymyxins. Analysing the treatment frequency, we found that a reduction in antimicrobial use in all types of pig production has occurred over time, accompanied by a rising percentage of farms without any usage. The lists of the World Health Organisation, World Organisation for Animal Health, and European Medicine Agency classify different antimicrobial substances as critically important. The vast differences between the respective weighted treatment frequencies allocated to the antimicrobials of main interest reflect the huge impact of the three categorisation systems. We concluded that, with the aim of creating national treatment guidelines supporting veterinarians to make treatment decisions, the list of the European Medicine Agency is the most suitable.

Canine real-time detection of SARS-CoV-2 infections in the context of a mass screening event.

INTRODUCTION: Previous research demonstrated that medical scent detection dogs have the ability to distinguish SARS-CoV-2 positive from negative samples with high diagnostic accuracy. To deploy these dogs as a reliable screening method, it is mandatory to examine if canines maintain their high diagnostic accuracy in real-life screening settings. We conducted a study to evaluate the performance of medical scent detection dogs under real-life circumstances. METHODS: Eight dogs were trained to detect SARS-CoV-2 RT-qPCR-positive samples. Four concerts with a total of 2802 participants were held to evaluate canines' performance in screening individuals for SARS-CoV-2 infection. Sweat samples were taken from all participants and presented in a line-up setting. In addition, every participant had been tested with a SARS-CoV-2 specific rapid antigen test and a RT-qPCR and they provided information regarding age, sex, vaccination status and medical disease history. The participants' infection status was unknown at the time of canine testing. Safety measures such as mask wearing and distance keeping were ensured. RESULTS: The SARS-CoV-2 detection dogs achieved a diagnostic specificity of 99.93% (95% CI 99.74% to 99.99%) and a sensitivity of 81.58% (95% CI 66.58% to 90.78%), respectively. The overall rate of concordant results was 99.68%. The majority of the study population was vaccinated with varying vaccines and vaccination schemes, while several participants had chronic diseases and were under chronic medication. This did not influence dogs' decisions. CONCLUSION: Our results demonstrate that SARS-CoV-2 scent detection dogs achieved high diagnostic accuracy in a real-life scenario. The vaccination status, previous SARS-CoV-2 infection, chronic disease and medication of the participants did not influence the performance of the dogs in detecting the acute infection. This indicates that dogs provide a fast and reliable screening option for public events in which high-throughput screening is required.

H7N9 avian influenza virus infection in men is associated with testosterone depletion.

Human infections with H7N9 avian influenza A virus that emerged in East China in 2013 and caused high morbidity rates were more frequently detected in men than in women over the last five epidemic waves. However, molecular markers associated with poor disease outcomes in men are still unknown. In this study, we systematically analysed sex hormone and cytokine levels in males and females with laboratory-confirmed H7N9 influenza in comparison to H7N9-negative control groups as well as laboratory-confirmed seasonal H1N1/H3N2 influenza cases (n = 369). Multivariable analyses reveal that H7N9-infected men present with considerably reduced testosterone levels associated with a poor outcome compared to non-infected controls. Regression analyses reveal that testosterone levels in H7N9-infected men are negatively associated with the levels of several pro-inflammatory cytokines, such as IL-6 and IL-15. To assess whether there is a causal relationship between low testosterone levels and avian H7N9 influenza infection, we used a mouse model. In male mice, we show that respiratory H7N9 infection leads to a high viral load and inflammatory cytokine response in the testes as well as a reduction in pre-infection plasma testosterone levels. Collectively, these findings suggest that monitoring sex hormone levels may support individualized management for patients with avian influenza infections.

Diagnosis and Prevalence of Chagas Disease in an Indigenous Population of Colombia.

Chagas disease (CD) is one of the leading neglected tropical diseases. In the Americas, CD is endemic in about 21 countries, but only less than 1% of the patients have access to medical treatment. Indigenous populations are particularly affected because they live in socio-economic and climate conditions that favor CD infections. In this study, diagnostic strategies and regional prevalence of the Chagas disease were assessed. In nine villages of the indigenous tribe Wiwa, 1134 persons were tested with a Chagas-antibody-specific rapid test (RT), two different Chagas-antibody-specific ELISAs and a Chagas-specific real-time polymerase chain reaction. The overall prevalence of CD in the villages was 35.4%, with a variation from 24.9% to 52.8% for the different communities. Rapid tests and ELISAs showed the same results in all cases. The proportion of replication-active infections, defined by positive PCR results, was 8.7%. In conclusion, the assessed indigenous population in Colombia was shown to be severely affected by CD. For a serological diagnosis, one rapid test was shown to be sufficient. Replacements of ELISAs by RT would decrease costs, increase feasibility and would relevantly help detect positive patients, especially if combined with the applied real-time PCR protocol. Real-time PCR can be considered for the detection of acute cases, outbreaks, chronic cases with re-infection/activation, as well as for therapy management and control.

Temporal Patterns of Phenotypic Antimicrobial Resistance and Coinfecting Pathogens in Glaesserella parasuis Strains Isolated from Diseased Swine in Germany from 2006 to 2021.

Glaesserella parasuis (Gps) causes high economic losses in pig farms worldwide. So far no vaccine provides cross-protection for different serotypes, so antibiotic treatment is widely used to cope with this pathogen. In this study, routine diagnostic data from 2046 pigs with Gps related diseases sent for necropsy to a German laboratory in the time period 2006-2021 were analysed retrospectively. In the time period 2018-2021, the most frequent serotypes (ST) detected were ST4 (30%) and ST13 (22%). A comparison of the reference period 2006-2013 prior to obligatory routine recording of antimicrobial usage in livestock with the period 2014-2021 resulted in a statistically significant decrease of frequencies of resistant Gps isolates for ceftiofur, enrofloxacin, erythromycin, spectinomycin, tiamulin and tilmicosin. While in 2006-2013 all isolates were resistant for tetracyclin and cephalothin, frequencies of resistant isolates decreased in the second time period to 28% and 62%, respectively. Parallel to the reduction of antimicrobial usage, during recent years a reduction in resistant Gps isolates has been observed, so only a low risk of treatment failure exists. Most frequently, pigs positive for Gps were also positive for S.suis (25.4%), PRRSV-EU (25.1%) and influenza virus (23%). The viral pathogens may act as potential trigger factors.

Prevalence of Common Diseases in Indigenous People in Colombia.

The Indigenous tribe called the Wiwa lives retracted in the Sierra Nevada de Santa Marta, Colombia. Little is known about their health status and whether the health care system in place covers their needs. In 2017 and 2018, a permanent physician was in charge for the Wiwa. Diseases and complaints were registered, ranked, and classified with the ICD-10 coding. Datasets from the Indigenous health care provider Dusakawi, collected from local health points and health brigades travelling sporadically into the fields for short visits, were compared. Furthermore, a list of provided medication was evaluated regarding the recorded needs. The most common complaints found were respiratory, infectious and parasitic, and digestive diseases. The top ten diagnoses collected in the health points and in the health brigade datasets were similar, although with a different ranking. The available medication showed a basic coverage only, with a critical lack of treatment for many severe, chronic, and life-threatening diseases. Most of the detected diseases in the Indigenous population are avoidable by an improvement in health care access, an expansion of the provided medication, and an increase in knowledge, hygiene, and life standards.

Evaluation of a Treadmill-Based Submaximal Fitness Test in Pugs, and Collecting Breed-Specific Information on Brachycephalic Obstructive Airway Syndrome.

Despite efforts of veterinarians and breeders, brachycephalic obstructive airway syndrome (BOAS) is still a common problem in pugs, underlining the need for objective tests to identify and prevent breeding with affected dogs. In the current study, a submaximal, treadmill-based fitness test was evaluated as a tool to identify signs of airway obstruction not recognisable under rest conditions. In addition to this, different body conformation and measurements were assessed regarding their association with BOAS. A total of 62 pugs and 10 mesocephalic dogs trotted with an individual comfort speed on a treadmill for 15 min. Before and during the examination, dogs were examined for signs of respiratory distress, and a functional BOAS grading was applied. The influence of body conformation on BOAS grading was tested in a univariable and multivariable logistic regression model. During exercise, more respiratory noises were observed, and existing respiratory noises became more apparent in comparison to when at rest. In the multivariable logistic regression model, no factor had a statistically significant influence on BOAS classification. Submaximal fitness testing helped to identify signs of respiratory distress not apparent under resting conditions, and could be a valuable addition for identifying dogs with BOAS. Performing testing on a treadmill facilitates continuous observation of the patients, and enables standardisation of the test regarding the test environment, as well as provides an uninterrupted, steady workload.