Sero-epidemiological investigation and cross-neutralization activity against SARS-CoV-2 variants in cats and dogs, Thailand.

Epidemiological data on SARS-CoV-2 infection in companion animals have been thoroughly investigated in many countries. However, information on the neutralizing cross-reactivity against SARS-CoV-2 variants in companion animals is still limited. Here, we explored the neutralizing antibodies against SARS-CoV-2 in cats and dogs between May 2020 and December 2021 during the first wave (a Wuhan-Hu-1-dominant period) and the fourth wave (a Delta-dominant period) of the Thailand COVID-19 outbreak. Archival plasma samples of 1,304 cats and 1,795 dogs (total = 3,099) submitted for diagnosis and health checks were collected at the Prasu-Arthorn Veterinary Teaching Hospital, Faculty of Veterinary Science, Mahidol University, Nakhon Pathom. A microneutralization test was used to detect neutralizing antibodies against the ancestral Wuhan-Hu-1 and the Delta variants. A plasma sample with neutralizing titers ≥10 was considered positive. Our results showed relatively low seroprevalence with seropositive samples detected in 8 out of 3,099 individuals (0.26, 95% CI 0.11-0.51%). Among these cases, SARS-CoV-2 neutralizing antibodies from both the ancestral Wuhan-Hu-1 and the Delta variants were found in three out of eight cases in two cats (n = 2) and one dog (n = 1). Furthermore, neutralizing antibodies specific to only the ancestral Wuhan-Hu-1 variant were exclusively found in one cat (n = 1), while antibodies against only the Delta variant were detected in four dogs (n = 4). Additionally, the neutralizing cross-activities against SARS-CoV-2 variants (Alpha, Beta, and Omicron BA.2) were observed in the seropositive cats with limited capacity to neutralize the Omicron BA.2 variant. In summary, the seropositivity among cats and dogs in households with an unknown COVID-19 status was relatively low in Thailand. Moreover, the neutralizing antibodies against SARS-CoV-2 found in the seropositive cats and dogs had limited or no ability to neutralize the Omicron BA.2 variant. Thus, monitoring SARS-CoV-2 infection and sero-surveillance, particularly in cats, is imperative for tracking virus susceptibility to the emergence of new SARS-CoV-2 variants.

Prevalence and characterization of antimicrobial-resistant Escherichia coli isolated from veterinary staff, pets, and pet owners in Thailand.

BACKGROUND: Companion animals may act as antimicrobial resistance (AMR) reservoirs. This study investigated the prevalence and AMR patterns of Escherichia coli in pets and people in close contact with pets. METHODS: A total of 955 samples were collected from veterinary clinics across Thailand by rectal and skin or ear swabs from dogs and cats and fecal swabs from veterinarians, veterinary assistants, and pet owners. The minimum inhibitory concentrations (MICs) of the obtained isolates were investigated using Sensititre™ MIC plates against 21 different antimicrobial drugs. RESULTS: Escherichia coli from pets was frequently resistant to ampicillin (100%) and amoxicillin-clavulanic acid (100%), whereas E. coli from pet owners, veterinarians, and veterinary assistants was mostly resistant to tetracycline. The multiple antibiotic resistance index revealed that multidrug-resistant E. coli isolates were frequently found in dogs (34.92%), cats (62.12%), veterinarians (61.11%), veterinarian assistants (36.36%), and pet owners (47.62%). The most common AMR genes identified in this study were blaCTX-M, blaTEM, tetA, and tetB, which were associated with the antimicrobial susceptibility results. Additionally, extended-spectrum beta-lactamase (ESBL)-associated genes (i.e., blaCTX-M, blaTEM, and blaSHV) were found in 21.69%, 71.97%, 27.78%, and 21.43% of E. coli isolated from dogs, cats, veterinarians, and pet owners, respectively. CONCLUSIONS: Our findings demonstrated the presence of AMR genes, particularly ESBL-associated genes, in E. coli isolated from healthy pets and veterinarians. This implies that these sources of E. coli could potentially be reservoirs for antibiotic resistance, thereby increasing the risk of harm to both humans and animals. These findings highlight the importance of implementing effective AMR control measures in veterinary practices, as bacteria resistant to commonly used antimicrobials are present in humans and animals.

Stakeholder attitudes and perspectives on wildlife disease surveillance as a component of a One Health approach in Thailand.

Coordinated wildlife disease surveillance (WDS) can help professionals across disciplines effectively safeguard human, animal, and environmental health. The aims of this study were to understand how WDS in Thailand is utilized, valued, and can be improved within a One Health framework. An online questionnaire was distributed to 183 professionals (55.7% response rate) across Thailand working in wildlife, marine animal, livestock, domestic animal, zoo animal, environmental, and public health sectors. Twelve semi-structured interviews with key professionals were then performed. Three-quarters of survey respondents reported using WDS data and information. Sectors agreed upon ranking disease control (76.5% of respondents) as the most beneficial outcome of WDS, while fostering new ideas through collaboration was valued by few participants (2.0%). Accessing data collected by one's own sector was identified as the most challenging (50%) yet least difficult to improve (88.3%). Having legal authority to conduct WDS was the second most frequently identified challenge. Interviewees explained that legal documentation required for cross-institutional collaborations posed a barrier to efficient communication and use of human resources. Survey respondents identified allocation of human resources (75.5%), adequate budget (71.6%), and having a clear communication system between sectors (71.6%) as highest priority areas for improvement to WDS in Thailand. Authorization from administrative officials and support from local community members were identified as challenges during in-person interviews. Future outreach may be directed toward these groups. As 42.9% of marine health professionals had difficulty knowing whom to contact in other sectors and 28.4% of survey respondents indicated that communication with marine health professionals was not applicable to their work, connecting the marine sector with other sectors may be prioritized. This study identifies priorities for addressing current challenges in the establishment of a general WDS system and information management system in Thailand while presenting a model for such evaluation in other regions.

The Agreement between Radiography and Fluoroscopy as Diagnostic Tools for Tracheal Collapse in Dogs.

Tracheal collapse is a common disease in small, middle-aged dogs, and imaging tools are essential for its diagnosis. Radiography and fluoroscopy are the main diagnostic modalities used, but their agreement in different regions is not well documented. In this study, the agreement between thoracic radiography and fluoroscopy in tracheal collapse was investigated in 29 dogs. The results showed that radiography detected a lower degree of collapsing trachea compared to fluoroscopy at the carina region (p < 0.001). However, there was no significant difference observed between the degree of collapsing trachea detected by radiography and fluoroscopy at the cervical, thoracic inlet, and intra-thoracic regions (p = 0.780, 0.537, and 0.213, respectively). The kappa statistic indicated a slight agreement at the cervical region at a 16.4% cut-off (κ = 0.20), while the other regions showed a non-agreement. In conclusion, although radiography is useful for screening, fluoroscopy was able to detect the degree of the collapsing trachea greater than radiography in the carina region. Additionally, if a collapse in the cervical region is detected by radiography, it is prone to have a positive relationship with fluoroscopy as well.

Antidiabetic effects of Andrographis paniculata supplementation on biochemical parameters, inflammatory responses, and oxidative stress in canine diabetes.

Introduction: Diabetes mellitus is a common endocrine disorder that causes hyperglycemia in dogs. Persistent hyperglycemia can induce inflammation and oxidative stress. This study aimed to investigate the effects of A. paniculata (Burm.f.) Nees (Acanthaceae) (A. paniculata) on blood glucose, inflammation, and oxidative stress in canine diabetes. A total of 41 client-owned dogs (23 diabetic and 18 clinically healthy) were included in this double-blind, placebo-controlled trial. Methods: The diabetic dogs were further divided into two treatments protocols: group 1 received A. paniculata extract capsules (50 mg/kg/day; n = 6) or received placebo for 90 days (n = 7); and group 2 received A. paniculata extract capsules (100 mg/kg/day; n = 6) or received a placebo for 180 days (n = 4). Blood and urine samples were collected every month. No significant differences in fasting blood glucose, fructosamine, interleukin-6, tumor necrosis factor-alpha, superoxide dismutase, and malondialdehyde levels were observed between the treatment and placebo groups (p > 0.05). Results and Discussion: The levels of alanine aminotransferase, alkaline phosphatase, blood urea nitrogen, and creatinine were stable in the treatment groups. The blood glucose levels and concentrations of inflammatory and oxidative stress markers in the client-owned diabetic dogs were not altered by A. paniculata supplementation. Furthermore, treatment with this extract did not have any adverse effects on the animals. Non-etheless, the effects of A. paniculata on canine diabetes must be appropriately evaluated using a proteomic approach and involving a wider variety of protein markers.

Utilizing MALDI-TOF MS and LC-MS/MS to access serum peptidome-based biomarkers in canine oral tumors.

Tumors frequently found in dogs include canine oral tumors, either cancerous or noncancerous. The bloodstream is an important route for tumor metastasis, particularly for late-stage oral melanoma (LOM) and late-stage oral squamous cell carcinoma (LOSCC). The present study aimed to investigate serum peptidome-based biomarkers of dogs with early-stage oral melanoma, LOM, LOSCC, benign oral tumors, chronic periodontitis and healthy controls, using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and liquid chromatography tandem mass spectrometry. A principal component analysis plot showed distinct clusters among all groups. Four peptides were identified, including peptidyl-prolyl cis-trans isomerase FKBP4 isoform X2 (FKBP4), steroid hormone receptor ERR1 (ESRRA or ERRA), immunoglobulin superfamily member 10 (IGSF10) and ATP-binding cassette subfamily B member 5 (ABCB5). FKBP4, ESRRA and ABCB5 were found to be overexpressed in both LOM and LOSCC, whereas IGSF10 expression was markedly increased in LOSCC only. These four proteins also played a crucial role in numerous pathways of cancer metastasis and showed a strong relationship with chemotherapy drugs. In conclusion, this study showed rapid screening of canine oral tumors using serum and MALDI-TOF MS. In addition, potential serum peptidome-based biomarker candidates for LOM and LOSCC were identified.

Use of proposed systolic and myocardial performance indices derived from simultaneous ECG and PCG recordings to assess cardiac function in healthy Beagles.

Background and Aim: Cardiac time intervals (CTIs) can provide important information on the electrical and mechanical properties of the heart. We hypothesized that cardiac function can be described using the combined power of electrocardiography (ECG) and phonocardiography (PCG) signals. This study aimed to (1) validate a novel custom device in measuring CTI parameters; (2) compare CTI parameters with a commercially available device and standard transthoracic echocardiography (STE); and (3) compare calculated systolic performance index (SPI) and myocardial performance index (MPI) with Tei index from the STE. Materials and Methods: This study determined CTIs based on simultaneous ECG and PCG recordings in 14 healthy Beagle dogs using the custom-built device. These CTI parameters were compared with a commercially available device (Eko DUO ECG + Digital Stethoscope; Eko DUO) and the STE. Agreement of CTI parameters between the custom device and the commercially available device or STE was evaluated. Calculated SPI and MPI based on Wigger's diagram were proposed, compared with SPI and Tei index, and correlated with STE parameters. Results: We found that the ECG and PCG parameters measured from the custom-built device did not differ from the commercially available device and the STE. By combining ECG and PCG signals, we established CTI parameters in healthy dogs including indices for systolic function (SPI: QS1/S1S2) and global cardiac function {F1 ([QS1+S2]/S1S2), F2 ([RS1+S2]/S1S2), and F3 (RS1 + [QS2-QT]/S1S2)}. The SPI, F2, and F3 were comparable with echocardiographic parameters describing systolic (Pre-ejection period/left ventricular ejection time [LVET]) and Tei index ([MCOdur-LVET]/LVET), respectively. Only SPI and F3 were correlated significantly with MCOdur and heart rate, respectively. Conclusion: We have validated the use of the custom-built device to describe CTIs that are comparable to the commercially available device and STE in healthy Beagles. The proposed SPI and MPI derived from CTI parameters can be useful in clinical practice to describe the cardiac function, especially in areas where access to STE is constrained.

Curcuminoid supplementation in canine diabetic mellitus and its complications using proteomic analysis.

Introduction: Inflammation and oxidative stress contribute to diabetes pathogenesis and consequences. Therapeutic approaches for canine diabetes remain a challenge. Curcumin has anti-inflammatory and anti-oxidative effects and is beneficial for humans with diabetes mellitus (DM); however, data on its impact on canine diabetes is limited. This study aimed to evaluate the potential for causing adverse effects, anti-inflammatory effects, anti-oxidative effects and proteomic patterns of curcuminoid supplementation on canine DM. Methods: Altogether, 18 dogs were divided into two groups: DM (n = 6) and healthy (n = 12). Curcuminoid 250 mg was given to the DM group orally daily for 180 days. Blood and urine sample collection for hematological parameters, blood biochemistry, urinalysis, oxidative stress parameters, inflammatory markers and proteomics were performed every 6 weeks. Results and discussion: Curcuminoid supplementation with standard therapy significantly decreased oxidative stress with the increased glutathione/oxidized glutathione ratio, but cytokine levels were unaffected. According to the proteomic analysis, curcuminoid altered the expression of alpha-2-HS-glycoprotein, transthyretin, apolipoprotein A-I and apolipoprotein A-IV, suggesting that curcuminoid improves insulin sensitivity and reduces cardiovascular complications. No negative impact on clinical symptoms, kidneys or liver markers was identified. This study proposed that curcuminoids might be used as a targeted antioxidant strategy as an adjunctive treatment to minimize diabetes complications in dogs.

Peptide barcodes in dogs affected by mitral valve disease with and without pulmonary hypertension using MALDI-TOF MS and LC-MS/MS.

Mitral valve disease (MVD) is an important and most frequently acquired heart disease found in dogs. MVD is classified into different stages according to its severity. There is a challenge in differentiation between asymptomatic and symptomatic stages of the MVD. Moreover, pulmonary hypertension (PH) is a common complication in dogs affected by MVD. In clinical practice, there are also some limitations to identify PH. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a technique that can characterize specific patterns of peptide mass called peptide barcodes from various samples. Besides, in combination with liquid chromatography-tandem mass spectrometry (LC-MS/MS), potential peptide sequences associated with specific conditions could be identified. The present study aimed to use MALDI-TOF coupled with LC-MS/MS to characterize specific peptide barcodes and potential peptide candidates in serum samples from healthy dogs, dogs with MVD stage B (MVD B, asymptomatic stage), MVD stage C (MVD C, symptomatic stage), MVD stage B with PH (MVD B PH), and MVD stage C with PH (MVD C PH). Discrete clusters of the 5 sample groups were identified by 3D plot analysis. Peptide barcodes also revealed differences in peptide patterns among the 5 groups. Six amino acid sequences of peptide candidates at 1,225.60, 1,363.85, 1,688.71, 1789.52, 2020.21, and 2156.42 Da were identified as part of the proteins CLCN1, CLUL1, EDNRA, PTEN, SLC39A7, and CLN6, respectively. The network interactions between these discovered proteins and common cardiovascular drugs were also investigated. These results demonstrate that MALDI-TOF MS has promise as an optional technique for diagnosing dogs affected by asymptomatic and symptomatic stages of MVD with and without PH. Further studies are required to identify peptide barcodes in dogs with other diseases to create peptide barcode databases in veterinary medicine before using this method as a novel diagnostic tool in the future.

One Health drivers of antibacterial resistance: Quantifying the relative impacts of human, animal and environmental use and transmission.

OBJECTIVES: Antibacterial resistance (ABR) is a major global health security threat, with a disproportionate burden on lower-and middle-income countries (LMICs). It is not understood how 'One Health', where human health is co-dependent on animal health and the environment, might impact the burden of ABR in LMICs. Thailand's 2017 "National Strategic Plan on Antimicrobial Resistance" (NSP-AMR) aims to reduce AMR morbidity by 50% through 20% reductions in human and 30% in animal antibacterial use (ABU). There is a need to understand the implications of such a plan within a One Health perspective. METHODS: A model of ABU, gut colonisation with extended-spectrum beta-lactamase (ESBL)-producing bacteria and transmission was calibrated using estimates of the prevalence of ESBL-producing bacteria in Thailand. This model was used to project the reduction in human ABR over 20 years (2020-2040) for each One Health driver, including individual transmission rates between humans, animals and the environment, and to estimate the long-term impact of the NSP-AMR intervention. RESULTS: The model predicts that human ABU was the most important factor in reducing the colonisation of humans with resistant bacteria (maximum 65.7-99.7% reduction). The NSP-AMR is projected to reduce human colonisation by 6.0-18.8%, with more ambitious targets (30% reductions in human ABU) increasing this to 8.5-24.9%. CONCLUSIONS: Our model provides a simple framework to explain the mechanisms underpinning ABR, suggesting that future interventions targeting the simultaneous reduction of transmission and ABU would help to control ABR more effectively in Thailand.