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Background Tick-borne pathogens are understudied among domestic animals in sub-Saharan Africa but represent significant threats to the health of domestic animals and humans. Specifically, additional data are needed on tick-borne pathogens in Chad, Africa. Surveillance was conducted among domestic dogs in Chad for selected tick-borne pathogens to measure (1) the prevalence of antibodies against Anaplasma spp., Borrelia burgdorferi , and Ehrlichia spp.; (2) the prevalence of infections caused by Hepatozoon spp., Ehrlichia canis , Anaplasma platys , and Babesia spp.; and (3) associations of pathogens with demographic, spatial, and temporal factors. Blood samples were collected from domestic dogs at three time points (May 2019, November 2019, June 2020) across 23 villages in southern Chad. Results Of the 428 dogs tested with the IDEXX SNAP 4Dx test in May 2019, 86% ( n  = 370, 95% CI = 83–90%) were positive for antibodies to Ehrlichia spp., 21% ( n  = 88, 95% CI = 17–25%) were positive for antibodies to Anaplasma spp., and 0.7% ( n  = 3, 95% CI = 0.1–2%) were positive for antibodies to Borrelia burgdorferi . Four different pathogens were detected via PCR. Hepatozoon spp. were most commonly detected (67.2–93.4%, depending on the time point of sampling), followed by E. canis (7.0-27.8%), A. platys (10.1–22.0%), and Babesia vogeli (0.4–1.9%). Dogs were coinfected with up to three pathogens at a single time point, and coinfections were most common in May 2019 compared to November 2019 and May 2020. Conclusions Overall, this study provides new data about the epidemiology of tick-borne pathogens in domestic dogs in Chad, with potential implications for dog and human health.

Background Disorders of the locomotor system in dogs, such as amputations or malformations, can be not only physically but also emotionally distressing. In this context, advances in medical and technological sciences offer tools and options with the aim of improving the quality of life of animals with locomotor problems. This case report aims to describe the custom development of a 3D exoprosthesis for a dog with an amputated hind limb. Case presentation A female dog, mixed breed, approximately 5 years old, was admitted to the Veterinary Clinic of the Centro Universitário de João Pessoa—Brazil, with locomotor problems due to low amputation of the left hind limb, without pain or sensitivity to touch on the amputated stump. Measurements of the amputated limb were collected to create a virtual model of the 3D exoprosthesis in a socket model. After simulations and tests, the prosthesis was materialized by 3D printing in collaboration with the Brazilian company 3D Medicine©, using polylactic acid (PLA) as the main material, an organic, lightweight, and resistant synthetic thermoplastic. The exoprosthesis was covered with protective material and fixed to the animal with a compressive bandage. Immediately after fixation, the animal demonstrated support of the limb on the prosthesis while standing, better distribution of body weight and relief of load on the contralateral limb. The increase in the time of use of the prosthesis was gradual and under supervision, after four weeks the dog did not present major difficulties in walking, running, and eating, in addition, no injuries to the amputated stump were observed. Veterinary physiotherapeutic follow-up was recommended. Conclusion This case report describes the development of a 3D exoprosthesis for dogs as a cost-effective option to reduce the locomotor impacts of limb amputation and improve quality of life. Techniques using additive manufacturing and 3D technology have immense potential for medical application, especially in veterinary medicine due to the difference in anatomical and body structure between domestic and wild animals.

Background Small extracellular vesicles (sEVs) derived from mesenchymal stem cells (MSCs) are recognized for their therapeutic potential in immune modulation and tissue repair, especially in veterinary medicine. This study introduces an innovative sequential stimulation (IVES) technique, involving low-oxygen gas mixture preconditioning using in vitro fertilization gas (IVFG) and direct current electrical stimulation (ES20), to enhance the anti-inflammatory properties of sEVs from canine adipose-derived MSCs (cAD-MSCs). Initial steps involved isolation and comprehensive characterization of cAD-MSCs, including morphology, gene expression, and differentiation potentials, alongside validation of the electrical stimulation protocol. IVFG, ES20, and IVES were applied simultaneously with a control condition. Stimulated cAD-MSCs were evaluated for morphological changes, cell viability, and gene expressions. Conditioned media were collected and purified for sEV isolation on Day1, Day2, and Day3. To validate the efficacy of IVES for sEV production, various analyses were conducted, including microscopic examination, surface marker assessment, zeta-potential measurement, protein quantification, nanoparticle tracking analysis, and determination of anti-inflammatory activity. Results We found that IVES demonstrated non-cytotoxicity and induced crucial genotypic changes associated with sEV production in cAD-MSCs. Interestingly, IVFG influenced cellular adaptation, while ES20 induced hypoxia activation. By merging these stimulations, IVES enhanced sEV stability and quality profiles. The cAD-MSC-derived sEVs exhibited anti-inflammatory activity in lipopolysaccharide-induced RAW264.7 macrophages, emphasizing their improved effectiveness without cytotoxicity or immunogenicity. These effects were consistent across day 3 collection, indicating the establishment of an effective protocol for sEV production. Conclusions This research established an innovative sequential stimulation method with positive impact on sEV characteristics including stability, quality, and anti-inflammatory activity. This study not only contributes to the enhancement of sEV production but also sheds light on their functional aspects for therapeutic interventions.

Canine mammary gland tumors (MGT) have a poor prognosis in intact female canines, posing a clinical challenge. This study aimed to establish novel canine mammary cancer cell lines from primary tumors and characterize their cellular and molecular features to find potential therapeutic drugs. The MGT cell lines demonstrated rapid cell proliferation and colony formation in an anchorage-independent manner. Vimentin and α-SMA levels were significantly elevated in MGT cell lines compared to normal canine kidney (MDCK) cells, while CDH1 expression was either significantly lower or not detected at all, based on quantitative real-time PCR (qRT-PCR) analysis. Functional annotation and enrichment analysis revealed that epithelial-mesenchymal transition (EMT) phenotypes and tumor-associated pathways, particularly the PI3K/Akt signaling pathway, were upregulated in MGT cells. BYL719 (Alpelisib), a PI3K inhibitor, was also examined for cytotoxicity on the MGT cell lines. The results show that BYL719 can significantly inhibit the proliferation of MGT cell lines in vitro. Overall, our findings suggest that the MGT cell lines may be valuable for future studies on the development, progression, metastasis, and management of tumors.

Background Microbial colonization during early life has a pivotal impact on the host health, shaping immune and metabolic functions, but little is known about timing and features of this process in dogs. The objectives of this study were to characterize the first step of intestinal microbiota development in naturally delivered canine puppies and to investigate its relationship with the maternal bacterial flora, using traditional culture and molecular analyses. Sixty puppies of two breeds, Appenzeller Cattle Dog ( n  = 3 dams) and Lagotto Romagnolo ( n  = 6), housed in the same breeding kennel, were included in the study. Swabs were collected in duplicate (for culture and for molecular analysis) from the dams’ vagina and rectum at the end of parturition, from puppies’ rectum, before maternal care, and from the environment (floor of the nursery and parturition box). Results 93.3% meconium samples showed bacterial growth, limited to a few colonies in 57.0% of cases. High growth was detected for Enterococcus faecalis , which was the most frequently isolated bacterium. The genus Enterococcus was one of the most represented in the dams’ rectum and vagina (88.9% and 55.6%, respectively). The genera Staphylococcus , Enterococcus , Escherichia and Proteus were also often isolated in meconium but were usually present in maternal samples as well, together with ubiquitous bacteria ( Acinetobacter , Psychrobacter ). In the environmental samples, just a few bacterial species were found, all with low microbial load. Additionally, bacteria of the phyla Proteobacteria, Firmicutes, and Actinobacteria were identified in meconium through molecular analysis, confirming the culture results and the early colonization of the newborn gut. Maternal, meconium and environmental samples had similar alpha diversity, while beta-diversity showed differences among families (i.e. a dam and her litter), and association indexes revealed a significant correlation between family members and between sample origin, suggesting a strong contribution of the maternal flora to the initial seeding of the canine neonatal gut and a strong individual dam imprint. Conclusion This study showed that the meconium of vaginally delivered puppies has its own microbiota immediately after birth, and that it is shaped by the dam, which gives a specific imprint to her litter.

Background Iron-deficiency is associated with increased morbidity and mortality in non-communicable diseases. However, iron parameters are rarely assessed in dogs. Here, we aimed to assess and correlate iron parameters in dogs suffering from Canine Atopic Dermatitis (CAD) compared to non-atopic, healthy dogs. Results For this retrospective study, blood values and sera of 34 dogs with confirmed CAD were compared with 94 healthy non-atopic dogs. In our cohort, dogs with CAD had significantly lower mean corpuscular volume (MCV, ) mean corpuscular hemoglobin (MCH) but higher white blood cell counts due to increased levels of circulating neutrophils and monocytes. CAD patients also had elevated total protein and c-reactive protein (CRP), but lower albumin levels compared to our healthy control dogs, indicated low-grade inflammation in the CAD cohort. Spearman correlations associated negatively clinical symptom (CADESI-4/PVAS) with MCV; ceruloplasmin and hepcidin, but positively with serum iron. Only in the CAD-cohort, MCV, CRP and albumin-levels negatively affected serum iron-levels and were positively associated with ceruloplasmin. Linear regression analysis revealed that serum iron-levels in CAD subjects, were positively dependent on hematocrit (packed cell volume, PCV) and albumin, and negatively dependent with white blood cells and neutrophils numbers. In contrast, in the healthy cohort, hepcidin was the sole factor associated with serum iron. Conclusions A decreased iron status was associated with a higher symptom burden. Iron homeostasis differed markedly in healthy and atopic dermatitis dogs. CAD patients had depleted iron-stores and presented themselves with subclinical inflammation.

Background Periodontal disease, an inflammatory condition initiated by the build-up of plaque on the tooth surface, is a common problem seen within veterinary practices. There are an increasing number of studies which indicate distinctive microbial profiles associated with healthy gingiva and periodontal disease. Most have been based on analysis of samples from populations of mixed breed dogs collected at a specific point in time. A study investigating the development of periodontal disease in Yorkshire terriers provided an opportunity to investigate the concurrent changes in the oral microbiota in this specific breed. Results Analysis of 42 subgingival plaque samples from 22 dogs (1 to 4 samples per dog) by 454 pyrosequencing of the V1-V3 region of the 16S rRNA gene resulted in 796,091 filtered sequence reads which were assigned to 286 operational taxonomic units (excluding those deemed noise). Statistical analysis showed that health and mild gingivitis were associated with higher relative abundance of taxa belonging to the phylum Proteobacteria (e.g., Moraxella and Pasteurellaceae). In moderate gingivitis there was increased representation of taxa belonging to the phylum Firmicutes (e.g., Peptostreptococcaceae, Lachnospiraceae, Erysipelotrichaceae and Frigovirgula ) and Bacteroidetes ( Porphyromonas canoris ). Periodontitis was also associated with an increased representation of some taxa belonging to the phylum Firmicutes (e.g., Peptosteptococcaceae), Spirochaetea (e.g., Treponema) and Synergistetes (e.g., Synergistales). Conclusions This study further advances our understanding of the bacterial changes associated with early periodontal disease. These can be leveraged to improve disease diagnosis, drive awareness and support recommendations for effective preventative and management strategies.

Background While the urogenital microbiota has recently been characterized in healthy male and female dogs, the influence of sex hormones on the urogenital microbiome of bitches is still unknown. A deeper understanding of the cyclic changes in urinary and vaginal microbiota would allow us to compare the bacterial populations in healthy dogs and assess the impact of the microbiome on various urogenital diseases. Therefore, the aim of this study was to characterize and compare the urogenital microbiota during different phases of the estrous cycle in healthy female dogs. DNA extraction, 16 S rDNA library preparation, sequencing and informatic analysis were performed to determine the vaginal and urinary microbiota in 10 healthy beagle dogs at each phase of the estrous cycle. Results There were no significant differences in alpha and beta diversity of the urinary microbiota across the different cycle phases. Similarly, alpha diversity, richness and evenness of vaginal bacterial populations were not significantly different across the cycle phases. However, there were significant differences in vaginal beta diversity between the different cycle phases, except for between anestrus and diestrus. Conclusion This study strongly suggests that estrogen influences the abundance of the vaginal microbiota in healthy female dogs, but does not appear to affect the urinary microbiome. Furthermore, our data facilitate a deeper understanding of the native urinary and vaginal microbiota in healthy female dogs.

Background Colorectal proliferative lesions are not common in dogs. However, recently we have observed an increase in the number of diagnosed cases and a lack of publications providing current epidemiological data on lesions of the large intestine in dogs. The aim of this study was a retrospective analysis of 217 canine colorectal non-neoplastic and neoplastic nodular lesions, and assessment of the frequency of occurrence of individual lesions and whether there is a risk of their occurrence depending on age, sex, or dog breed. Results Half of the cases (52.5%) were malignant tumours with a significant predominance of adenocarcinoma (42.9%). In the group of malignant non-epithelial lesions, lymphoma and sarcomas predominated (4.1% and 4.1%, respectively) followed by three cases of plasmacytoma. Benign neoplastic tumours constituted almost one-third of all cases (26.7%) with obvious dominance of adenoma (24.0%). Benign mesenchymal tumours were represented only by leiomyoma (2.8%). The non-neoplastic lesions were represented by a heterogeneous group of polyps (20.3%) with a slight advantage of hyperplastic type (9.7%) and less numerous inflammatory, fibroblastic, lymphoid, and hamartomatous polyps. One case of ganglioneuromatosis in hamartomatous polyp was diagnosed. The vast majority of lesions, both non-neoplastic and neoplastic, were found in the rectum. French Bulldogs were the most numerous breeds in our study, especially among adenomas. Furthermore, benign tumours were diagnosed in younger animals than malignant tumours. Conclusions The results of our research provided new data expanding knowledge about the epidemiology of colorectal neoplastic and non-neoplastic proliferative lesions in dogs. Our results indicate that the majority of colorectal proliferative lesions in dogs are malignant, which is alarming. French Bulldogs could possibly be predisposed to proliferative lesions of the large intestine, and this predisposition was statistically confirmed in adenomas. Moreover, benign tumours may occur in animals as young as 1–2 years old.

Background Quality of life is an essential component of decision-making in veterinary oncology. Poor management of adverse events during chemotherapy can impair dogs’ quality of life and be life-threatening. Consequently, client-reported outcome measures (CROMs) are being proposed to help assess both clinical signs and quality of life. The purpose of this rapid review was to: (1) identify existing CROMs that have been used to assess dogs’ clinical signs and quality of life during chemotherapy; and (2) evaluate their methodological development to inform adaptation or development of a CROM for use in routine clinical practice, including remote monitoring. Databases (Scopus, Web of Science, PUBMED/MEDLINE) were searched for CROMs (questionnaires) completed by a non-expert family member about their companion dog. CROM content (domains measured) and scale quality were appraised. Results Ten CROMs were identified and three were variations of the same tool. Content of the CROMs varied considerably (range 3–17 domains) with gastrointestinal being the most frequently measured clinical sign cluster (9/10 studies), followed by mobility/ambulatory activity (7/10) and global quality of life (6/10). No CROMs adhered to quality standards for the development of questionnaires and most failed to include qualitative design methods and basic psychometric assessment to ensure reliability and validity (such as internal consistency, test-retest reliability or factor analysis). Conclusion The validity and reliability of existing chemotherapy CROMs for dogs remains under-tested. Although CROMs combined with remote digital monitoring have the potential to enhance patient care, as has been demonstrated with physician-based oncology, there is a need to apply quality standards to ensure optimal validation. Interdisciplinary collaborations would likely improve CROM quality and clinical utility thereby allowing veterinary healthcare professionals to better support their patients.