Mesenchymal stem cells: A promising antimicrobial therapy in veterinary medicine.

Growing antimicrobial resistance (AMR) is a threat to human and animal populations citing the limited available options. Alternative antimicrobial options or functional enhancement of currently available antimicrobials remains only options. One of the potential options seems stem cells especially the mesenchymal stem cells (MSCs) that show antimicrobial properties. These cells additionally have pro-healing effects that may plausibly improve healing outcomes. MSCs antimicrobial actions are mediated either through direct cell-cell contact or their secretome that enhances innate immune mediated antimicrobial activities. These cells synergistically enhance efficacy of currently available antimicrobials especially against the biofilms. Reciprocal action from antimicrobials on the MSCs functionality remains poorly understood. Currently, the main limitation with MSCs based therapy is their limited efficacy. This demands further understanding and can be enhanced through biotechnological interventions. One of the interventional options is the 'priming' to enhance MSCs resistance and specific expression potential. The available literature shows potential antimicrobial actions of MSCs both ex vivo as well as in vivo. The studies on veterinary species are very promising although limited by number and extensiveness in details for their utility as standard therapeutic agents. The current review aims to discuss the role of animals in AMR and the potential antimicrobial actions of MSCs in veterinary medicine. The review also discusses the limitations in their utilization as standard therapeutics.

Mesenchymal stem cell basic research and applications in dog medicine.

The stem cells, owing to their special characteristics like self-renewal, multiplication, homing, immunomodulation, anti-inflammatory, and dedifferentiation are considered to carry an "all-in-one-solution" for diverse clinical problems. However, the limited understanding of cellular physiology currently limits their definitive therapeutic use. Among various stem cell types, currently mesenchymal stem cells are extensively studied for dog clinical applications owing to their readily available sources, easy harvesting, and ability to differentiate both into mesodermal, as well as extramesodermal tissues. The isolated, culture expanded, and characterized cells have been applied both at preclinical as well as clinical settings in dogs with variable but mostly positive results. The results, though positive, are currently inconclusive and demands further intensive research on the properties and their dependence on the applications. Further, numerous clinical conditions of dog resemble to that of human counterparts and thus, if proved rewarding in the former may act as basis of therapy for the latter. The current review throws some light on dog mesenchymal stem cell properties and their potential therapeutic applications.

Mesenchymal stem cell: Basic research and potential applications in cattle and buffalo.

Characteristic features like self-renewal, multilineage differentiation potential, and immune-modulatory/anti-inflammatory properties, besides the ability to mobilize and home distant tissues make stem cells (SCs) a lifeline for an individual. Stem cells (SCs) if could be harvested and expanded without any abnormal change may be utilized as an all-in-one solution to numerous clinical ailments. However, slender understanding of their basic physiological properties, including expression potential, behavioral alternations during culture, and the effect of niche/microenvironment has currently restricted the clinical application of SCs. Among various types of SCs, mesenchymal stem cells (MSCs) are extensively studied due to their easy availability, straightforward harvesting, and culturing procedures, besides, their less likelihood to produce teratogens. Large ruminant MSCs have been harvested from various adult tissues and fetal membranes and are well characterized under in vitro conditions but unlike human or other domestic animals in vivo studies on cattle/buffalo MSCs have mostly been aimed at improving the animals' production potential. In this document, we focused on the status and potential application of MSCs in cattle and buffalo.

Adult Stem Cell Research in Light of the Bovine Mammary Gland Regenerative Medicine.

The bovine mammary gland has vital importance in the dairy sector, as it is considered a source of basic dairy product, milk. Mammary gland affections are widespread, which affect the dairy industry economically and pose a potential public health hazard. Current therapeutic options are ineffective in controlling the infection and regenerating the gland effectively. Antimicrobials commonly used against mastitis make their way into the milk . In order to find a solution to these problems, advanced therapeutic options, like the one for stem cells, are considered. Mammary gland stem cells (MaSCs) are considered to maintain tissue homeostasis. The characterization of these cells and their derived lineages (progenitor cells and mammary epithelial cells) may potentially provide the physiological status or production potential of the gland. However, current isolation methods are cumbersome and fall short to isolate a pure line of cattle MaSCs from progenitors or other differentiated epithelial cells. An alternative to the therapeutic application of MaSCs is the mesenchymal stem cell (MSC). These cells can potentially control microbial infection, show anti-inflammatory and other pro-healing effects, and furthermore enhance mammary epithelial cell secretory potential to ensure tissue regeneration. The current review focuses on MaSCs and MSCs properties in light of the bovine mammary gland regeneration.

Extracellular vesicles derived from mesenchymal stem cells - a novel therapeutic tool in infectious diseases.

Extracellular vesicles (EVs) are nano-sized lipid-bilayer encapsulated vesicles produced by the cells. These EVs are released into the surrounding space by almost all cell types. The EVs help in intercellular communication via their payloads which contain various proteins, lipids, and nucleic acids generated from the donor cells and allow for synergistic responses in surrounding cells. In recent years, EVs have been increasingly important in treating infectious diseases, including respiratory tract infections, urinary tract infections, wound infections, sepsis, and intestinal infections. Studies have confirmed the therapeutic value of mesenchymal stem cell-derived EVs (MSC-EVs) for treating infectious diseases to eliminate the pathogen, modulate the resistance, and restore tissue damage in infectious diseases. This can be achieved by producing antimicrobial substances, inhibiting pathogen multiplication, and activating macrophage phagocytic activity. Pathogen compounds can be diffused by inserting them into EVs produced and secreted by host cells or by secreting them as microbial cells producing EVs carrying signalling molecules and DNA shielding infected pathogens from immune attack. EVs play a key role in infectious pathogenesis and hold great promise for developing innovative treatments. In this review, we discuss the role of MSC-EVs in treating various infectious diseases.

Epithelial to mesenchymal transition in mammary gland tissue fibrosis and insights into drug therapeutics.

Background: The epithelial-mesenchymal transition (EMT) is a multi-step morphogenetic process in which epithelial cells lose their epithelial properties and gain mesenchymal characteristics. The process of EMT has been shown to mediate mammary gland fibrosis. Understanding how mesenchymal cells emerge from an epithelial default state will aid in unravelling the mechanisms that control fibrosis and, ultimately, in identifying therapeutic targets to alleviate fibrosis. Methods: The effects of EGF and high glucose (HG) on EMT in mammary epithelial cells, MCF10A and GMECs, as well as their pathogenic role, were studied. In-silico analysis was used to find interacting partners and protein-chemical/drug molecule interactions. Results: On treatment with EGF and/or HG, qPCR analysis showed a significant increase in the gene expression of EMT markers and downstream signalling genes. The expression of these genes was reduced on treatment with EGF+HG combination in both cell lines. The protein expression of COL1A1 increased as compared to the control in cells treated with EGF or HG alone, but when the cells were treated with EGF and HG together, the protein expression of COL1A1 decreased. ROS levels and cell death increased in cells treated with EGF and HG alone, whereas cells treated with EGF and HG together showed a decrease in ROS production and apoptosis. In-silico analysis of protein-protein interactions suggest the possible role of MAPK1, actin alpha 2 (ACTA2), COL1A1, and NFκB1 in regulating TGFß1, ubiquitin C (UBC), specificity protein 1 (SP1) and E1A binding protein P300 (EP300). Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment suggests advanced glycation end products-receptor for advanced glycation end products (AGE-RAGE) signalling pathway, relaxin signalling pathway and extra cellular matrix (ECM) receptor interactions underlying fibrosis mechanism. Conclusion: This study demonstrates that EGF and HG induce EMT in mammary epithelial cells and may also have a role in fibrosis.

Mesenchymal stem cells derived from bone marrow and adipose tissue for repairing acute sciatic nerve injury in a rabbit model.

Peripheral nerve injury is one of the common disabling clinical conditions and around 50% of the cases end up in permanent impairment. Due to the lack of effective treatment options regenerative medicine employing stem cells is being evaluated. The presented study evaluated and compared regeneration potential of mesenchymal stem cells (MSCs) derived from bone marrow (BM) and adipose tissue (AD) in acute rabbit sciatic nerve injury (axonotmesis) model. A total of n = 54 grey giant rabbits were made subject of the study and divided equally into 3 groups: Control, BM-MSCs in Collagen I and AD-MSCs in Collagen I as per the treatment given. Iliac crest BM and omental AD was harvested from the same donor for isolation and culture of MSCs. The repair of sciatic nerve injury was evaluated on days 60 and 90. The clinical and histopathological scores and SEM morphology was better in cell treated groups as compared to the control. Morphology and histological studies revealed injured nerve in different levels of regenerative process. Gene expression was more than double for N-Cadherin in cell treated groups as compared to the control, especially at day 60. Between cell treated groups, BM-MSCs group showed better response as compared to the AD-MSCs, although statistically non-significant (p > 0.05). Incomplete nerve regeneration observed under various diagnostic parameters was in compliance to the incomplete clinical recovery at day 90. It was concluded that MSCs may improve sciatic nerve healing but fall short of complete regeneration at day 90, although BM-MSCs may have an edge over AD-MSCs.

Cryopreserved allogeneic bone marrow mesenchymal stem cells show better osteochondral defect repair potential than adipose tissue mesenchymal stem cells.

Mesenchymal stem cells (MSCs) due to their characteristic properties have a potential to treat osteoarthritis, one of the major growing joint problems. MSCs show differential ex vivo chondrogenic potential on the basis of source that remains to be validated under in vivo environment. This study compared chondrogenic potential of MSCs derived from two common sources, adipose tissue (AD) and bone marrow (BM) under ex vivo and in vivo environments. The randomized placebo controlled osteochondral defect (OCD) study divided n = 72 rabbits equally into Control, AD-MSCs and BM-MSCs groups. Ex vivo chondrogenic induction resulted in an increased aggrecan fold expression in BM-MSCs and AD-MSCs. The former cell type had significantly (p<0.05) higher fold expression as compared to the latter. The cell treated OCDs had significantly reduced gene expression for inflammatory markers (IL-6, IL-8 and TNF-α) as compared to the control. In OCD study, radiography, MRI, gross observation, histopathology and SEM revealed that the cell treated defects were early filled by the tissue that had better surface architecture and matrices as compared to the control. BM-MSCs treated defects had better scores especially for gross and histopathology than the AD-MSCs. Gene expression for osteochondral regulation and cartilaginous matrices was higher in BM-MSCs group while only for matrices including the Col I in AD-MSCs as compared to the control. It was concluded that OCD in the cell treated groups are filled early with mostly a fibrocartilaginous to hyaline tissue. BM-MSCs may have an edge over AD-MSCs in OCD repair.

Clinical applications of adipose-derived stromal vascular fraction in veterinary practice.

Adipose tissue-derived stromal vascular fraction (AdSVF) comprises a heterogeneous cell population, including the multipotent mesenchymal stem cells, hematopoietic stem cells, immune cells, endothelial cells, fibroblasts, and pericytes. As such, multipotent adipose tissue-derived mesenchymal stem cells (AdMSCs), are one of the important components of AdSVF. Commonly used techniques to harvest AdSVF involve enzymatic or non-enzymatic methods. The enzymatic method is considered to be the gold standard technique due to its higher yield. The cellular components of AdSVF can be resuspended in normal saline, platelet-rich plasma, or phosphate-buffered saline to produce a ready-to-use solution. Freshly isolated AdSVF has exhibited promising osteogenic and vasculogenic capacity. AdSVF has already been proven to possess therapeutic potential for osteoarthritis management. It is also an attractive therapeutic option for enhancing wound healing. In addition, the combined use of AdSVF and platelet-rich plasma has an additive stimulatory effect in accelerating wound healing and can be considered an alternative to AdMSC treatment. It is also widely used for managing various orthopaedic conditions in clinical settings and has the potential for regenerating bone, cartilage, and tendons. Autologous AdSVF cells are used along with bone substitutes and other biological factors as an alternative to conventional bone grafting techniques owing to their promising osteogenic and vasculogenic capacity. It can also be used for treating osteonecrosis, meniscus tear, chondromalacia, and tendon injuries in veterinary practice. It has several advantages over in vitro expanded AdMSC, including precluding the need for culturing, reduced risk of cell contamination, and cost-effectiveness, making it ideal for clinical use.

Ultrasound, Dacryocystorhinography and Morphological Examination of Normal Eye and Lacrimal Apparatus of the Donkey (Equus asinus).

The study investigated normal macromorphological and ultrasonographic features of the eye and lacrimal gland, as well as normal dacryocystorhinography of the donkey (Equus asinus) in Egypt. A total of 36 donkeys of different ages, weights, and sexes were included in the study: 21 live animals for ultrasonography and dacryocystorhinography, and 15 cadaver skulls for morphological anatomy of the lacrimal apparatus. The ultrasound biometric values of the eye were 33.7 ± 1.7 mm for axial globe length (AGL), 39.8 ± 2.1 mm for globe diameter (GD), 10.8 ± 0.7 mm for lens thickness (LT), 3.2 ± 0.7 mm for anterior chamber depth (ACD), and 19.3 ± 1.6 mm for vitreous chamber depth (VCD). The lacrimal gland was recognized as a hypoechogenic structure with an anechoic core, located at the dorsolateral aspect of the orbit, and ovoid in shape. The mean NLD length was 193.0 ± 9.8 mm by radiography and 206.0 ± 20.4 mm by gross assessment. One NL orifice (NLO) was noticed on each side, with a diameter of 3.0 ± 0.1 mm and located 12.1 ± 2.1 mm from the dorsal commissure of the nostril. These results may act as the baseline for proper management of conditions of the eye and lacrimal apparatus in the donkey in the future.

Cryopreserved allogeneic mesenchymal stem cells enhance wound repair in full thickness skin wound model and cattle clinical teat injuries.

The study was aimed to evaluate and compare the healing potential of mesenchymal stem cells (MSCs) derived from two common sources (iliac crest derived bone marrow and omental fat) in a full thickness skin wound model. Bone marrow derived MSCs clinical efficacy in the repair of cattle teat fistulae (cutaneous and muco-cutaneous wounds) was also evaluated. In a completely randomized placebo controlled experimental full thickness skin wound model, n=36 were randomly divided into three equal groups: groups I, II and III receiving Phosphate buffered saline (PBS), BM-MSCs and adipose tissue MSCs (AD-MSCs), respectively. Grossly early reduction in inflammation and enhanced epithelialization in the cell-treated groups as compared to the control was seen. Microscopy, ultramicroscopy, gene expression analysis and mechanical testing revealed better and early matrix formation with a reduced scar formation and a higher tensile strength in the cell-treated groups as compared to the control. An overall comparable healing in the cell treated groups was observed, although BM-MSCs had led to the better matrix formation tending to scarless healing while the AD-MSCs had led to the early wound closure with a good tissue strength. In the case controlled bovine clinical teat injuries study (n=17) repaired surgically, BM-MSCs (n=13) or PBS (n=4) was injected locally. In surgico-MSCs treated cases, 84.6% non-recurrence rate was observed as compared to the 50% seen in the control. It was concluded that MSCs irrespective of the donor tissue have potential to improve healing of full thickness cutaneous wounds and/ fistulae.

Advances in therapeutic and managemental approaches of bovine mastitis: a comprehensive review.

Mastitis (intramammary inflammation) caused by infectious pathogens is still considered a devastating condition of dairy animals affecting animal welfare as well as economically incurring huge losses to the dairy industry by means of decreased production performance and increased culling rates. Bovine mastitis is the inflammation of the mammary glands/udder of bovines, caused by bacterial pathogens, in most cases. Routine diagnosis is based on clinical and subclinical forms of the disease. This underlines the significance of early and rapid identification/detection of etiological agents at the farm level, for which several diagnostic techniques have been developed. Therapeutic regimens such as antibiotics, immunotherapy, bacteriocins, bacteriophages, antimicrobial peptides, probiotics, stem cell therapy, native secretory factors, nutritional, dry cow and lactation therapy, genetic selection, herbs, and nanoparticle technology-based therapy have been evaluated for their efficacy in the treatment of mastitis. Even though several strategies have been developed over the years for the purpose of managing both clinical and subclinical forms of mastitis, all of them lacked the efficacy to eliminate the associated etiological agent when used as a monotherapy. Further, research has to be directed towards the development of new therapeutic agents/techniques that can both replace conventional techniques and also solve the problem of emerging antibiotic resistance. The objective of the present review is to describe the etiological agents, pathogenesis, and diagnosis in brief along with an extensive discussion on the advances in the treatment and management of mastitis, which would help safeguard the health of dairy animals.

Allogeneic mesenchymal stem cells and growth factors in gel scaffold repair osteochondral defect in rabbit.

Aim: An attempt was made to improve osteochondral healing with allogeneic mesenchymal stem cells (MSCs) along with certain growth factors. Materials & methods: Induced knee osteochondral defects were filled as: phosphate buffer saline (group A); MSCs in collagen gel (group B); group B plus insulin like growth factor-1 (group C); group C plus transforming growth factor ß-1 (group D). Results: Gross and scanning electron microscopy showed superior morphology and surface architecture of the healed tissue in groups D and C. Histologically, group D revealed hyaline cartilage characteristic features followed in order by group C and group B. In all treatment groups, chondrogenic matrix, collagen II2B (col II 2B) and aggrecan were secreted. Conclusion: Combined use of MSCs and growth factors could accelerate osteochondral healing.

Equine Mesenchymal Stem Cells: Properties, Sources, Characterization, and Potential Therapeutic Applications.

Properties like sustained multiplication and self-renewal, and homing and multilineage differentiation to undertake repair of the damaged tissues make stem cells the lifeline for any living system. Therefore, stem cell therapy is regarded to carry immense therapeutic potential. Though the dearth of understanding about the basic biological properties and pathways involved in therapeutic benefits currently limit the application of stem cells in humans as well as animals, there are innumerable reports that suggest clinical benefits of stem cell therapy in equine. Among various stem cell sources, currently adult mesenchymal stem cells (MSCs) are preferred for therapeutic application in horse owing to their easy availability, capacity to modulate inflammation, and promote healing. Also the cells carry very limited teratogenic risk compared to the pluripotent stem cells. Mesenchymal stem cells were earlier considered mainly for musculoskeletal tissues, but now may also be utilized in other diverse clinical problems in horse, and the results may be extrapolated even for human medicine. The current review highlights biological properties, sources, mechanisms, and potential therapeutic applications of stem cells in equine practice.

Animal mesenchymal stem cell research in cartilage regenerative medicine - a review.

Healing of articular cartilage is a major clinical challenge as it also lacks a direct vasculature and nerves, and carries a limited number of resident chondrocytes that do not proliferate easily. Damaged articular cartilages are usually replaced by fibrocartilages, which are mechanically and structurally weaker and less resilient. Regenerative medicine involving stem cells is considered to have a definitive potential to overcome the limitations associated with the currently available surgical methods of cartilage repair. Among various stem cell types, mesenchymal stem cells (MSCs) are preferred for clinical applications. These cells can be readily derived from various sources and have the ability to trans-differentiate into various tissue-specific cells, including those of the cartilage by the process of chondrogenesis. Compared to embryonic or induced pluripotent stem cells (iPSCs), no ethical or teratogenic issues are associated with MSCs. These stem cells are being extensively evaluated for the treatment of joint affections and the results appear promising. Unlike human medicine, in veterinary medicine, the literature on stem cell research for cartilage regeneration is limited. This review, therefore, aims to comprehensively discuss the available literature and pinpoint the achievements and limitations associated with the use of MSCs for articular cartilage repair in animal species.

Mesenchymal Stem Cell Research in Veterinary Medicine.

BACKGROUND: The researchers working in the field of medical, veterinary and other biological sciences have uniformly described unique characteristic features of stem cells including selfrenewal, prolonged multiplication, immuno-modulation and multi-lineage differentiation. These characteristics of stem cells have opened new horizons in cellular therapy for the management of numerous incurable diseases in human and veterinary patients. Though the mechanisms involved in reparative processes may be similar, the indication for the stem cell therapy may be disparate in human and veterinary subjects. OBJECTIVE: Among various stem cell types, currently mesenchymal stem cells (MSCs) are extensively studied in regenerative medicine owing to their readily available sources, easy harvesting and ability to differentiate both into mesodermal as well as non-mesodermal tissues under specific culture conditions with little associated ethical issues. The objective of the study was to analyze and summarize the studies and their results pertaining to the basic biology of stem cells, preclinical trials and their potential therapeutic application in veterinary medicine. RESULTS: MSCs have been variably isolated, cultured and characterized from almost all the body tissues and fetal membranes in domestic and pet animals. The cells have been studied in numerous pre-clinical and clinical studies, in addition, to lab animal models. The results although are promising but need further extensive research studies before the cellular application becomes a clinical reality in veterinary medicine. CONCLUSION: The current review throws some light on different aspects of mesenchymal stem cells like sources, isolation, characterization and their potential therapeutic applications in farm and pet animals.

Mesenchymal stem cells with IGF-1 and TGF- ß1 in laminin gel for osteochondral defects in rabbits.

OBJECTIVE: Healing of articular cartilage is still a challenge due to its limited potential to regenerate. In the present study, we evaluated allogenic bone marrow mesenchymal stem cells (BM-MSCs) alone or in combination with growth factors, insulin-like growth factor-1 (IGF-1) and transforming growth factor-ß1 (TGF-ß1) in laminin scaffolds for healing of osteochondral defects. DESIGN: Osteochondral defects of 4mm (diameter) x 5mm (depth) were induced in the rabbit knee joints and treated with phosphate-buffered saline (PBS; control), BM-MSCs, BM-MSCs in laminin, BM-MSCs in laminin with IGF-1, or BM-MSCs in laminin with IGF-1 and TGF-ß1 in 10 animals each. Gross, radiographic, scanning electron microscopic (SEM) and histologic examinations besides chondrocyte-specific genes expression by quantitative real time qPCR were carried out at 8 and 12 weeks. RESULTS: Gross and SEM examination revealed superior morphology and surface architecture of the healing site in animals that received MSCs with IGF-1 or IGF-1 and TGF-ß1. The application of laminin composites containing MSCs with IGF-1 and TGF-ß1 significantly enhanced hyaline cartilage formation with improved cellular arrangement, proteoglycan deposition, clear tidemark zone and subchondral bone formation. However, regenerated tissue in defects that received only MSCs had poor tidemark zone and proteoglycans deposition Aggrecan and Coll2 expression was significantly higher in case of MSCs with growth factors. CONCLUSION: The treatment with BM-MSCs combined with IGF-1/TGF-ß1 into laminin gel scaffold might enhance the restoration of hyaline cartilage in osteochondral defect.

Vertebral scale system to measure heart size in thoracic radiographs of Indian Spitz, Labrador retriever and Mongrel dogs.

AIM: To establish reference values of vertebral heart score (VHS) in Indian Spitz, Labrador retriever, and Mongrel dogs; to assess applicability of VHS in these three dog breeds; to determine if breed, recumbency side, gender, body weight, and thoracic depth (TD) to thoracic width (TW) ratio has an influence on the VHS measurement in these dog breeds. MATERIALS AND METHODS: A total of 60, client owned, clinically healthy Indian Spitz (n=20, mean age = 4.25±2.15 years, body weight = 11.87±2.7 kg), Labrador retriever (n=20, mean age = 4.75±1.91 years, body weight = 27.31±5.43 kg), and Mongrel dogs (n=20, mean age = 4.25±1.52 years, body weight = 16.25±3.99 kg), having no radiological and clinical signs of cardiovascular or pulmonary disease were included in the study. All dogs were restrained manually and left lateral (LL) and right lateral (RL) radiographic views were obtained. The size of heart in lateral radiographs was calculated using VHS method. Besides, the TD, TW and TD: TW were calculated to determine the type of thoracic conformation in the dog breeds. In addition to this, the effect of breed, side of recumbency, gender, body weight, and TD to TW ratio on the calculation of VHS was determined. RESULTS: VHS was calculated in all the animals of the breeds. VHS in Spitz and Labrador retriever was significantly (p<0.0001, p<0.0001, respectively) >9.7±0.5 v. RL and LL VHS in Mongrel dog was significantly (p<0.037) >9.7±0.5 v. Significant (p<0.05) differences in the VHS were observed among Spitz, Labrador retriever and Mongrel dogs, being higher for Labrador retriever followed by Spitz and Mongrel dogs. VHS in RL recumbency was significantly (p<0.001) greater than VHS in LL recumbency in all three breeds. LL and RL VHS correlated significantly with each other in Spitz (r=0.58; p=0.02), Labrador retriever (r=0.87; p<0.0001), and Mongrel dogs (r=0.93; p<0.0001). Significant (p<0.05) differences in the TD and TW were observed among Spitz, Labrador retriever, and Mongrel dogs. Non-significant effect of gender, body weight, and TD to TW ratio on the VHS measurement was observed in each dog breed. CONCLUSION: Breed-specific VHS reference ranges should be used for the objective measurement of heart size in dogs. Furthermore, the radiographic view should also be taken into consideration to avoid any erroneous interpretation of cardiac enlargement in dogs.

Reference values of six-limb-lead electrocardiogram in conscious Labrador retriever dogs.

Breed-wise standard electrocardiographic values in dogs have been reported as there is variation in body and chest conformation, which limits the application of data of one breed for other breed. Labrador retrievers being originated from hunting dogs, their electrocardiogram (ECG) values might be different from standard normal range of other dog breeds. So, the purpose of the present study was to determine the standard ECG of Labrador retrievers and to check effect of body weight, gender and breed upon different ECG parameters. Six-lead ECGs, three bipolar standard limb leads (I, II and III) and three augmented unipolar limb leads (aVR, aVL and aVF), were taken from 24 Labrador retrievers positioned in right lateral recumbency without any chemical restraint. Amplitude and duration of P wave and QRS complex, PR interval, QT interval and mean electrical axis and heart rate were measured in each recording. Non-significant effect of gender and body weight was seen on all the ECG waves. Deep Q waves in Leads I, II and aVF and variation in relation to QRS pattern were noted. It was concluded that retrievers had a specific shape of QRS complex which must be considered when evaluating a patient suspected of having cardiac disease. However, amplitude and durations of different ECG waves in all the six leads were statistically not affected by gender or body weight.