Meta-transcriptomic analysis of companion animal infectomes reveals their diversity and potential roles in animal and human disease.

Companion animals such as cats and dogs harbor diverse microbial communities that can potentially impact human health due to close and frequent contact. To better characterize their total infectomes and assess zoonotic risks, we characterized the overall infectomes of companion animals (cats and dogs) and evaluated their potential zoonotic risks. Meta-transcriptomic analyses were performed on 239 samples from cats and dogs collected across China, identifying 24 viral species, 270 bacterial genera, and two fungal genera. Differences in the overall microbiome and infectome composition were compared across different animal species (cats or dogs), sampling sites (rectal or oropharyngeal), and health status (healthy or diseased). Diversity analyses revealed that viral abundance was generally higher in diseased animals compared to healthy ones, while differences in microbial composition were mainly driven by sampling site, followed by animal species and health status. Disease association analyses validated the pathogenicity of known pathogens and suggested potential pathogenic roles of previously undescribed bacteria and newly discovered viruses. Cross-species transmission analyses identified seven pathogens shared between cats and dogs, such as alphacoronavirus 1, which was detected in both oropharyngeal and rectal swabs albeit with differential pathogenicity. Further analyses showed that some viruses, like alphacoronavirus 1, harbored multiple lineages exhibiting distinct pathogenicity, tissue, or host preferences. Ultimately, a systematic evolutionary screening identified 27 potential zoonotic pathogens in this sample set, with far more bacterial than viral species, implying potential health threats to humans. Overall, our meta-transcriptomic analysis reveals a landscape of actively transcribing microorganisms in major companion animals, highlighting key pathogens, those with the potential for cross-species transmission, and possible zoonotic threats. IMPORTANCE: This study provides a comprehensive characterization of the entire community of infectious microbes (viruses, bacteria, and fungi) in companion animals like cats and dogs, termed the "infectome." By analyzing hundreds of samples from across China, the researchers identified numerous known and novel pathogens, including 27 potential zoonotic agents that could pose health risks to both animals and humans. Notably, some of these zoonotic pathogens were detected even in apparently healthy pets, highlighting the importance of surveillance. The study also revealed key microbial factors associated with respiratory and gastrointestinal diseases in pets, as well as potential cross-species transmission events between cats and dogs. Overall, this work sheds light on the complex microbial landscapes of companion animals and their potential impacts on animal and human health, underscoring the need for monitoring and management of these infectious agents.

Penalization and Color Code-Technical Approaches for Methods' Greenness and Whiteness Appraisal in Veterinary Medication: Assay of Toltrazuril Suspension.

BACKGROUND: Intestinal coccidiosis is a debilitating disease in poultry and livestock, leading to economic impact worldwide. Coccidiosis is prevented and treated in broilers by the inclusion of anticoccidials in feed. Toltrazuril is administered in potable water to treat coccidiosis. OBJECTIVE: Three robust analytical methods for quantitation of toltrazuril in pure and pharmaceutical formulations are developed. Furthermore, ecological metrics; either penalization- or color-code-based techniques are applied for the appraisal of assays. METHODS: Firstly, Second-Derivative (Δλ; 5 nm) spectrophotometric method; Toltrazuril is measured from peak to peak at 244-260 nm within a linearity range of 5-25 µg/mL. The second one is a high-performance thin-layer chromatography (HPTLC) analysis performed on an aluminum sheet of silica gel using ethyl acetate, methanol, ammonium chloride buffer, and water (8:1:0.5:0.5) (%V/V) as the elution phase. Toltrazuril, at a retardation factor of 0.66 ± 0.01, is linearly determined in the range of 1-9 µg/spot at 243 nm. The third one is Reversed Phase-HPLC-diode array detection, using Agilent column C18 (5 µm, 4.6 x 150 mm) in isocratic elution mode with a mobile phase of acetonitrile and water in a ratio of 80:20 (v/v), respectively, at 1 mL/min flow rate. Toltrazuril elutes at a retention time of 2.58 ± 0.1 min and is linearly determined at 243 nm in the range of 0.25-25 µg/mL. RESULTS: Calculated 2D-values and peak areas are highly correlated to their corresponding drug concentrations at coefficients; r > 0.999. All methods were ICH validated and applied to dosage form with satisfactory % recoveries (97-103%). Statistical comparisons reported one using t-test and F-test disclose insignificant variation. Examining greenness and whiteness norms, proposed methods were evaluated and ranked alongside four different reported methods. CONCLUSION: The proposed methods are green, accurate, and can be applied in routine quality control for the determination of toltrazuril in pharmaceutical formulations.

Development of a novel scoring system to predict mortality in canine patients with infection.

OBJECTIVES: To develop a novel illness severity scoring system to identify canine patients with infection that are at higher risk of mortality. KEY FINDINGS: Mentation (Modified Glasgow Coma Scale and the Acute Patient Physiologic and Laboratory Evaluation mentation score), heart rate, pH, Pvco2, potassium, and total plasma protein were found to be associated with mortality on univariate analysis. Logistic regression found that mentation, heart rate, and Pvco2 combined to form the following score: [(100 × mentation score) + (3 × heart rate) + (10 × Pvco2], which was predictive of mortality (area under the receiver operating characteristic curve: 0.88). A cutoff of 900 provided a sensitivity of 80% and a specificity of 80%. If the cutoff was changed to 800, the sensitivity was 100% and the specificity was 50%. SIGNIFICANCE: A novel scoring system was developed to predict mortality in hospitalized dogs with confirmed or suspected infection. Further prospective evaluation in a larger patient cohort is necessary to validate this score.

A Rare Case of Streptococcus equi Subspecies Zooepidemicus Bacteremia.

Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) is a zoonotic pathogen that primarily infects horses, pigs, and dogs. Although rare, it has also been shown to infect humans who consume unpasteurized dairy food or have direct contact with horses. Here, we present a case of S. zooepidemicus bacteremia in a patient without a clear mode of transmission. An 86-year-old male with a past medical history of coronary artery disease, heart failure with reduced ejection fraction, complete heart block status post pacemaker, hypertension, hyperlipidemia, and type 2 diabetes mellitus presented to the Emergency Department with fever and chills. He had fevers and rigors for three days but denied weight loss, cough, sore throat, or rashes. In the Emergency Department, vital signs revealed a fever of 101.2 degrees Fahrenheit and a heart rate of 110 with other stable vital signs. The physical exam was unremarkable except for tachycardia, and laboratory work revealed no leukocytosis but elevated inflammatory markers and elevated lactate. Computed tomography of the chest, abdomen, and pelvis did not reveal any source of infection. Blood cultures grew S. zooepidemicus and the Infectious Diseases team was consulted, who started the patient on Penicillin G. Due to concern for pacer-lead infective endocarditis, transthoracic and transesophageal echocardiograms were performed, which did not show valvular vegetations. Repeat blood cultures showed clearance of the infection, and the patient was ultimately discharged on amoxicillin. While our patient denied consuming unpasteurized dairy products or having direct contact with horses, upon further questioning, he did endorse family members who occasionally interacted with horses. This case is valuable as it adds to the sparse literature on S. zooepidemicus infections specifically in humans. Extensive history taking is of utmost importance when a clear source of infection is not easily identifiable. Further research is also needed to better understand the various modes of transmission of this bacterium to better target and caution those at an increased risk of infection.

Ex vivo diffusion tensor imaging of the plantar nerves is feasible in the horse.

OBJECTIVE: The objective of this study was to optimize an MRI-based diffusion tensor imaging (DTI) protocol for imaging the plantar nerves at the level of the tarsus in normal equine limbs. SAMPLE: 12 pelvic cadaver limbs from horses without evidence of proximal suspensory pathology were imaged with a 3T MRI system. METHODS: For diffusion-weighted imaging, b values of 600, 800, and 1,000 s/mm2 were tested. Data were processed with DSI Studio. Cross-sectional areas of the medial and lateral plantar nerve along the plantar tarsus were recorded. The length and number of fiber tracts, signal-to-noise ratio, and DTI variables were recorded. RESULTS: At the level of interest, the mean cross-sectional areas of the plantar nerves ranged from 5.03 to 7.42 mm2. The DTI maps consistently generated tracts in the region of the lateral and medial plantar nerves with DTI values in the range of values reported for peripheral nerves in humans. Our findings demonstrate that DTI of the medial and lateral plantar nerves can be performed successfully and used to generate quantitative parameters including fractional anisotropy and mean, axial, and radial diffusivity. CLINICAL RELEVANCE: Quantitative data generated with this imaging technique can be used to noninvasively characterize the microstructural integrity of neural tissue with possible applications in the evaluation of pathologic changes to the plantar tarsal and metatarsal nerves of horses with proximal suspensory desmopathy.

Survey indicates addressing workplace environment, work-life balance, and flexibility are key to attracting and retaining veterinarians in academia.

OBJECTIVE: Referencing growing concerns over the recruitment and retention of faculty in academic veterinary medicine, the authors hypothesized that among surveyed veterinary residents and early-career faculty, work-life balance and workplace climate and culture are stronger motivators than financial considerations, regardless of demographic factors such as gender, race/ethnicity, and area of specialization. SAMPLE: 541 participants were included in data analysis. METHODS: A mixed methods approach was utilized, incorporating both quantitative data and qualitative, free-text responses to better understand veterinary career choices by contextualizing factors associated with academic medicine. RESULTS: Factors underpinning career-related decision-making were ranked by level of importance as (1) workplace environment/culture, (2) personal well-being/work-life balance, (3) salary and bonuses, (4) geographic location, (5) facilities and resources, (6) benefits, and (7) schedule flexibility. Desires for workload balance, schedule flexibility, support from leadership, and mentorship and collaboration were among the top themes of qualitative responses for both residents and early career faculty respondents. Factors influencing career decision-making for resident and early-career faculty are varied. Workplace environment, work-life balance, and schedule flexibility are areas that academic institutions can address and continue to improve and that are likely to positively impact entry into academia and the desire to stay. CLINICAL RELEVANCE: This study sought to understand factors related to career decision-making and interest in academic veterinary medicine among residents and early-career faculty. Understanding these factors can support efforts to recruit and retain faculty in academic veterinary medicine.

Clinical Presentation, Bacteriologic Findings and Possible Risk Factors for Ischemic Teat Necrosis in Cattle-A Case Series.

Ischemic teat necrosis (ITN) is a growing problem in the dairy industry characterized by teat lesions, necrosis, pruritus and automutilation. Despite the economic and welfare consequences, there is no treatment, and the etiology of the disease remains poorly understood. The aim of this study was to investigate ITN by analyzing its clinical presentation, potential risk factors and microbial involvement. Methods included collection of milk and swab samples from affected cows over a period of one-and-a-half years and completion of questionnaires by veterinarians and farmers. Microbial testing included PCR testing for Treponema spp. and cultural testing by anaerobic and aerobic incubation on blood agar. The results showed a high and significant prevalence of Treponema spp. and Staphylococcus aureus in affected teats compared to non-ITN-affected control teats, indicating their potential role in the development of ITN. Other factors such as edema and milking practices also appear to contribute to the tissue damage. First-lactation and early-lactation heifers are particularly at risk. In conclusion, ITN appears to have a multifactorial etiology with both infectious and non-infectious factors playing a role. Further research is needed to better understand the complex interplay of these factors and to develop effective prevention and management strategies.

Halicin: A New Horizon in Antibacterial Therapy against Veterinary Pathogens.

It is crucial to discover novel antimicrobial drugs to combat resistance. This study investigated the antibacterial properties of halicin (SU3327), an AI-identified anti-diabetic drug, against 13 kinds of common clinical pathogens of animal origin, including multidrug-resistant strains. Employing minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) assessments, halicin demonstrated a broad-spectrum antibacterial effect. Time-killing assays revealed its concentration-dependent bactericidal activity against Escherichia coli ATCC 25922 (E. coli ATCC 25922), Staphylococcus aureus ATCC 29213 (S. aureus ATCC 29213), and Actinobacillus pleuropneumoniae S6 (APP S6) after 4 h of treatment at concentrations above the MIC. Halicin exhibited longer post-antibiotic effects (PAEs) and sub-MIC effects (PA-SMEs) for E. coli 25922, S. aureus 29213, and APP S6 compared to ceftiofur and ciprofloxacin, the commonly used veterinary antimicrobial agents, indicating sustained antibacterial action. Additionally, the results of consecutive passaging experiments over 40 d at sub-inhibitory concentrations showed that bacteria exhibited difficulty in developing resistance to halicin. Toxicology studies confirmed that halicin exhibited low acute toxicity, being non-mutagenic, non-reproductive-toxic, and non-genotoxic. Blood biochemical results suggested that halicin has no significant impact on hematological parameters, liver function, and kidney function. Furthermore, halicin effectively treated respiratory A. pleuropneumoniae infections in murine models. These results underscore the potential of halicin as a new antibacterial agent with applications against clinically relevant pathogens in veterinary medicine.

Concentration-resistance relationship and PK/PD evaluation of danofloxacin against emergence of resistant Pasteurella multocida in an in vitro dynamic model.

AIMS: This study aimed to assess the pharmacokinetic/pharmacodynamic (PK/PD) targets of danofloxacin to minimize the risk of selecting resistant Pasteurella multocida mutants and to identify the mechanisms underlying their resistance in an in vitro dynamic model, attaining the optimum dosing regimen of danofloxacin to improve its clinical efficacy based on the mutant selection window (MSW) hypothesis. METHODS AND RESULTS: Danofloxacin at seven dosing regimens and 5 days of treatment were simulated to quantify the bactericidal kinetics and enrichment of resistant mutants upon continuous antibiotic exposure. The magnitudes of PK/PD targets associated with different efficacies were determined in the model. The 24 h area under the concentration-time curve (AUC) to minimum inhibitory concentration (MIC) ratios (AUC24h/MIC) of danofloxacin associated with bacteriostatic, bactericidal and eradication effects against P. multocida were 34, 52, and 64 h. This translates to average danofloxacin concentrations (Cav) over 24 h being 1.42, 2.17, and 2.67 times the MIC, respectively. An AUC/MIC-dependent antibacterial efficacy and AUC/mutant prevention concentration (MPC)-dependent enrichment of P. multocida mutants in which maximum losses in danofloxacin susceptibility occurred at a simulated AUC24h/MIC ratio of 72 h (i.e. Cav of three times the MIC). The overexpression of efflux pumps (acrAB-tolC) and their regulatory genes (marA, soxS, and ramA) was associated with reduced susceptibility in danofloxacin-exposed P. multocida. The AUC24h/MPC ratio of 19 h (i.e. Cav of 0.8 times the MPC) was determined to be the minimum mutant prevention target value for the selection of resistant P. multocida mutants. CONCLUSIONS: The emergence of P. multocida resistance to danofloxacin exhibited a concentration-dependent pattern and was consistent with the MSW hypothesis. The current clinical dosing regimen of danofloxacin (2.5 mg kg-1) may have a risk of treatment failure due to inducible fluoroquinolone resistance.

Becoming a matter of veterinary concern.

Horses in Great Britain are living into increasingly older age and are often regarded as friends or family members by their owner. The horse is reliant on their owner to meet their needs and this paper discusses how horse owners frame an issue that becomes a matter of veterinary concern within the context of the older horse. Qualitative methods were used to explore the experiences of owners and veterinarians. Data were collected and analysed using a grounded theory approach during the period 2019-2022. Analysis identified that owners undertook an ongoing and iterative process of assessment, monitoring and decision making in relation to the animal and any changes they observed. Matters that became a veterinary concern required the owner to formulate the issue as something that fell within the knowledge domain of the veterinarian. Veterinarians had a medicalised view of older horse health and their perspectives on socially acceptable care were shaped by their understanding of species-specific needs, and whether owners were providing appropriately for those needs. The formulation of a matter of veterinary concern was itself shaped by an owner's experiential knowledge of both veterinary matters and their horse. The extent to which owners felt like they and their individual horse mattered during interactions with veterinarians affected whether they adopted veterinary advice and the nature of future veterinary employment. Findings demonstrate how matters of health, disease, and the role of professionalised forms of medical knowledge, are not static but constantly changing and interacting over time. An issue that became a matter of veterinary concern was contextual, and rooted in individual relationships. The significance of veterinarian-owner interactions in shaping future consumption of veterinary health care may be underestimated.

Not just in man's best friend: A review of Staphylococcus pseudintermedius host range and human zoonosis.

Staphylococcus pseudintermedius is one species in the commensal staphylococcal population in dogs. While it is commonly carried on healthy companion dogs it is also an opportunistic pathogen associated with a range of skin, ear, wound and other infections. While adapted to dogs, it is not restricted to them, and we have reviewed its host range, including increasing reports of human colonisation and infections. Despite its association with pet dogs, S. pseudintermedius is found widely in animals, covering companion, livestock and free-living species of birds and mammals. Human infections, typically in immunocompromised individuals, are increasingly being recognised, in part due to improved diagnosis. Colonisation, infection, and antimicrobial resistance, including frequent multidrug resistance, among S. pseudintermedius isolates represent important One Health challenges.

Antimicrobial peptides in livestock: a review with a one health approach.

Antimicrobial peptides (AMPs), often referred to as nature's antibiotics, are ubiquitous in living organisms, spanning from bacteria to humans. Their potency, versatility, and unique mechanisms of action have garnered significant research attention. Unlike conventional antibiotics, peptides are biodegradable, adding to their appeal as potential candidates to address bacterial resistance in livestock farming-a challenge that has been under scrutiny for decades. This issue is complex and multifactorial, influenced by a variety of components. The World Health Organization (WHO) has proposed a comprehensive approach known as One Health, emphasizing the interconnectedness of human-animal-environment relationships in tackling such challenges. This review explores the application of AMPs in livestock farming and how they can mitigate the impact of this practice within the One Health framework.

Exotic Animal Practice in East Asia: A Retrospective Study of Species Distribution and Their Common Diseases at 2 Exotics-only Veterinary Clinics in the East Asia Region.

The objective of this clinical retrospective study was to analyze the prevalence and distribution of different avian and exotic animals presented to 2 exotics-only veterinary hospital in Hong Kong and Taiwan over a 1 year period. Exotic companion mammals, predominated by rabbits (Oryctolagus cuniculus) that were often diagnosed with fractures, were the most commonly presented group of patients in the hospital in Hong Kong while second most of that in Taiwan, with dental disease being commonly presented in the species. This study provided a general overview of avian and exotic patients presented to exotics-only practices in the East Asia region.

[Radiation therapy in veterinary medicine at the University of Liège]. / Radiothérapie en médecine vétérinaire à l'Université de Liège.

Radiation therapy has many indications in veterinary oncology and allows a multidisciplinary approach for the treatment of canine and feline patients. Radiation therapy can be recommended as a sole therapy in case of radiosensitive tumors or can be associated to surgery and/or chemotherapy after marginal excision for example. It can also be recommended as a palliative treatment for patients with an inoperable or painful tumor or disseminated disease. Radiation therapy significantly improves the quality of life and survival time of treated animals and should be part of the therapeutic modalities in veterinary medicine. The University of Liège developed the first veterinary center of radiation therapy in Belgium and can therefore participate in improving therapeutic management of cancerous animal patients.

La radiothérapie présente de nombreuses indications en oncologie vétérinaire et permet une approche multidisciplinaire pour le traitement de nos patients cancéreux canins et félins.La radiothérapie peut être recommandée seule pour le traitement de tumeurs radiosensibles, ou associée à la chirurgie et/ou la chimiothérapie notamment lors d'exérèse marginale. Elle a également sa place dans une prise en charge palliative de certains patients présentant une tumeur inopérable ou douloureuse, ou encore une maladie disséminée. La radiothérapie permet d'améliorer significativement la qualité et l'espérance de vie des animaux traités et à ce titre, doit faire partie de l'arsenal thérapeutique vétérinaire. L'Université de Liège possède, depuis peu, l'unique centre de radiothérapie vétérinaire en Belgique et peut ainsi participer à l'amélioration de la prise en charge des animaux cancéreux.

Corrigendum: How does chronic pain impact the lives of dogs: an investigation of factors that are associated with pain using the Animal Welfare Assessment Grid.

[This corrects the article DOI: 10.3389/fvets.2024.1374858.].

Bacteriophage-derived endolysins as innovative antimicrobials against bovine mastitis-causing streptococci and staphylococci: a state-of-the-art review.

Bacteriophage-encoded endolysins, peptidoglycan hydrolases breaking down the Gram-positive bacterial cell wall, represent a groundbreaking class of novel antimicrobials to revolutionize the veterinary medicine field. Wild-type endolysins exhibit a modular structure, consisting of enzymatically active and cell wall-binding domains, that enable genetic engineering strategies for the creation of chimeric fusion proteins or so-called 'engineered endolysins'. This biotechnological approach has yielded variants with modified lytic spectrums, introducing new possibilities in antimicrobial development. However, the discovery of highly similar endolysins by different groups has occasionally resulted in the assignment of different names that complicate a straightforward comparison. The aim of this review was to perform a homology-based comparison of the wild-type and engineered endolysins that have been characterized in the context of bovine mastitis-causing streptococci and staphylococci, grouping homologous endolysins with ≥ 95.0% protein sequence similarity. Literature is explored by homologous groups for the wild-type endolysins, followed by a chronological examination of engineered endolysins according to their year of publication. This review concludes that the wild-type endolysins encountered persistent challenges in raw milk and in vivo settings, causing a notable shift in the field towards the engineering of endolysins. Lead candidates that display robust lytic activity are nowadays selected from screening assays that are performed under these challenging conditions, often utilizing advanced high-throughput protein engineering methods. Overall, these recent advancements suggest that endolysins will integrate into the antibiotic arsenal over the next decade, thereby innovating antimicrobial treatment against bovine mastitis-causing streptococci and staphylococci.

A narrative review of the phenomenon of predatory journals to create awareness among researchers in veterinary medicine.

In recent years, especially since the COVID-19 pandemic, the number of predatory journals has increased significantly. Predatory journals exploit the "open-access model" by engaging in deceptive practices such as charging high publication fees without providing the expected quality and performing insufficient or no peer review. Such behaviors undermine the integrity of scientific research and can result in researchers having trouble identifying reputable publication opportunities, particularly early-career researchers who struggle to understand and establish the correct criteria for publication in reputable journals. Publishing in journals that do not fully cover the criteria for scientific publication is also an ethical issue. This review aimed to describe the characteristics of predatory journals, differentiate between reliable and predatory journals, investigate the reasons that lead researchers to publish in predatory journals, evaluate the negative impact of predatory publications on the scientific community, and explore future perspectives. The authors also provide some considerations for researchers (particularly early-career researchers) when selecting journals for publication, explaining the role of metrics, databases, and artificial intelligence in manuscript preparation, with a specific focus on and relevance to publication in veterinary medicine.

Dungeons and Dragons: Culture Differences in Attitudes Toward Exotic Animals.

The way exotic animals are viewed around the world differ. These views and attitudes are constantly evolving and may eventually converge in the way that cats and dogs are now widely accepted as part of most societies around the world. The choice and popularity of exotic animal species as pets is a complex interaction of many different factors, including legislation and availability. Media representation may be a factor too, but there has been very little evidence to demonstrate this. Regardless of the region, exotic animal practice appears to be an area of veterinary practice that is on the rise.

Salvia miltiorrhiza ameliorates endometritis in dairy cows by relieving inflammation, energy deficiency and blood stasis.

Introduction: According to traditional Chinese veterinary medicine, endometritis is caused by a combination of Qi deficiency, blood stasis, and external evil invasion. Salvia miltiorrhiza is a traditional Chinese medicine that counteracts blood stasis and has additional demonstrated effects in boosting energy and restraining inflammation. Salvia miltiorrhiza has been employed in many traditional Chinese prescriptions that have proven effective in healing clinical dairy cow endometritis. Methods: the in vivo effect of Salvia miltiorrhiza in treating endometritis was evaluated in dairy cows. In addition, bovine endometrial epithelium cell inflammation and rat blood stasis models were employed to demonstrate the crosstalk between energy, blood circulation and inflammation. Network analysis, western blotting, qRT-PCR and ELISA were performed to investigate the molecular mechanism of Salvia miltiorrhiza in endometritis treatment. Results: The results demonstrate that treatment with Salvia miltiorrhiza relieves uterine inflammation, increases blood ATP concentrations, and prolongs blood clotting times. Four of the six Salvia miltiorrhiza main components (SMMCs) (tanshinone IIA, cryptotanshinone, salvianolic acid A and salvianolic acid B) were effective in reversing decreased ATP and increased IL-1ß, IL-6, and IL-8 levels in an in vitro endometritis model, indicating their abilities to ameliorate the negative energy balance and external evil invasion effects of endometritis. Furthermore, in a blood stasis rat model, inflammatory responses were induced in the absence of external infection; and all six SMMCs inhibited thrombin-induced platelet aggregation. Network analysis of SMMC targets predicted that Salvia miltiorrhiza may mediate anti-inflammation via the Toll-like receptor signaling pathway; anti-aggregation via the Platelet activation pathway; and energy balance via the Thermogenesis and AMPK signaling pathways. Multiple molecular targets within these pathways were verified to be inhibited by SMMCs, including P38/ERK-AP1, a key molecular signal that may mediate the crosstalk between inflammation, energy deficiency and blood stasis. Conclusion: These results provide mechanistic understanding of the therapeutic effect of Salvia miltiorrhiza for endometritis achieved through Qi deficiency, blood stasis, and external evil invasion.

Characterization of the plasma proteome from healthy adult dogs.

Introduction: Bloodwork is a widely used diagnostic tool in veterinary medicine, as diagnosis and therapeutic interventions often rely on blood biomarkers. However, biomarkers available in veterinary medicine often lack sensitivity or specificity. Mass spectrometry-based proteomics technology has been extensively used in the analysis of biological fluids. It offers excellent potential for a more comprehensive characterization of the plasma proteome in veterinary medicine. Methods: In this study, we aimed to identify and quantify plasma proteins in a cohort of healthy dogs and compare two techniques for depleting high-abundance plasma proteins to enable the detection of lower-abundance proteins via label-free quantification liquid chromatography-mass spectrometry. We utilized surplus lithium-heparin plasma from 30 healthy dogs, subdivided into five groups of pooled plasma from 6 randomly selected individuals each. Firstly, we used a commercial kit to deplete high-abundance plasma proteins. Secondly, we employed an in-house method to remove albumin using Blue-Sepharose. Results and discussion: Among all the samples, some of the most abundant proteins identified were apolipoprotein A and B, albumin, alpha-2-macroglobulin, fibrinogen beta chain, fibronectin, complement C3, serotransferrin, and coagulation factor V. However, neither of the depletion techniques achieved significant depletion of highly abundant proteins. Despite this limitation, we could detect and quantify many clinically relevant proteins. Determining the healthy canine proteome is a crucial first step in establishing a reference proteome for canine plasma. After enrichment, this reference proteome can later be utilized to identify protein markers associated with different diseases, thereby contributing to the diagnosis and prognosis of various pathologies.