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ABSTRACT Blood samples from 200 female dogs divided into 6 groups (diestrus, mucometra, pyometra) were evaluated, with the pyometra group categorized according to the ASA classification (American Society of Anesthesiologists), that is, from ASA II to V, totaling six groups. Aiming to analyze the acute phase proteins (APP), hematological and serum biochemical profiles of the female dogs in the study, establishing the differential diagnosis and prognosis according to the group. The SDS-PAGE method was used for protein fractionation, complete blood count using an automated hematological analyzer and histogram, biochemical tests performed using a semi-automatic spectrophotometer and measurement of serum concentrations of sodium and ionic calcium using the selective ion method. The results showed that female dogs with pyometra categorized in the ASA II to V classification revealed biochemical alterations between the study groups, as well as the acute phase proteins, presented variations according to the systemic involvement, degree of inflammatory response and ASA classification of the female dogs with pyometra, in which ceruloplasmin (p=0.07) and transferrin (p=0.07) did not show statistical significance, but showed an increase in their concentration according to the inflammatory evolution, albumin (p

Definition the fertile period is one of the most important factors contributing to fertilization. Therefore, this analysis is important to solve the problem of infertility. The methods that available today do not provide the expected result, because the vast majority of unsuccessful mating result are related to the incorrect determination of the ovulation period. Therefore, it is important to assess the condition of the ovaries, and the prospect is ultrasound examination, which is an effective method of monitoring the reproductive organs. Ultrasound analysis is a non-invasive and effective method to establish the fertile period in bitches which makes it possible to track changes in the ovaries, detect follicles, their number, and determine the true (morphological) signs of ovulation. Clinical-visual and hormonal studies which are based on the indirect determination of the optimal period of female insemination have the advantages in compare with vaginal cytology. Particularly, the use of ultrasound in real time allows to visualize the morphological changes of the ovaries and uterus – the irregular shape of the ovaries with large (0.6-1.2 cm) anechoic structures of a rounded or oval shape with a thin capsule, thickening of the endometrium (up to 0.5-0.8 cm) with a hypoechoic structure of the uterine horns. However, in our research we encountered certain difficulties, which turned out to be the same as other authors. Echography of the ovaries can seem a bit complicated, because the follicles that persist in them have a picture similar to that before and after ovulation. Some ovulated follicles do not always completely collapse (fall) during ovulation and are gradually replaced by luteal tissue while maintaining an echogenic picture on the days of the next ovulation. Non-ovulated follicles can complicate sonography interpretation. To diagnose ovulation, an ultrasound analysis should be performed twice a day, which is inconvenient for the owners. But if the ultrasound test is carried out less often than aforementioned number, there is a risk to miss the moment of ovulation, because the follicles before and after ovulation are very similar. An even more significant problem is the location of the ovaries in bitches – costovertebral angle surrounded by fat and an ovarian sac, which can create technical difficulties. In addition, this diagnostic approach may be more difficult in fat or very large dogs, as well as in dogs with thick skin (Shar Pei, Newfoundland).

Five female dogs were presented at Veterinary Clinical Complex, College of Veterinary and Animal Sciences, Sardar Vallabh Bhai Patel University of Agriculture & Technology, Meerut, Uttar Pradesh with the history of protruded mass from the vagina. The animals were anorexic, dull with slightly elevated temperature. Confirmatory diagnosis was made by histopathological examination of these tissues. Two of the she dogs were diagnosed with vaginal myxoma and other three were diagnosed with fibroma. The treatment included surgical removal of protruded mass and follow-up treatment with antibiotics, NSAIDS, multi-mineral and multi-vitamin supplements. [ABSTRACT FROM AUTHOR]

Simple Summary: Breast cancer research is frequently performed using a variety of models, including animal models, in an attempt to provide information that can be translated into human clinical practice. This review aims to demonstrate the similarities and differences in the anatomy of the mammary glands of women, female dogs and female rats, as well as the epidemiology, risk factors, and histopathological features of breast/mammary cancer in each model. It also demonstrates the advantages and disadvantages of each model, as they can serve as sources for several in vitro models, further increasing the translational potential of these models. Breast cancer is one of the most common and well-known types of cancer among women worldwide and is the most frequent neoplasm in intact female dogs. Female dogs are considered attractive models or studying spontaneous breast cancer, whereas female rats are currently the most widely used animal models for breast cancer research in the laboratory context. Both female dogs and female rats have contributed to the advancement of scientific knowledge in this field, and, in a "One Health" approach, they have allowed broad understanding of specific biopathological pathways, influence of environmental factors and screening/discovery of candidate therapies. This review aims to clearly showcase the similarities and differences among woman, female dog and female rat concerning to anatomical, physiological and histological features of the mammary gland and breast/mammary cancer epidemiology, in order to better portray breast tumorigenesis, and to ensure appropriate conclusions and extrapolation of results among species. We also discuss the major aspects that stand out in these species. The mammary glands of female dogs and women share structural similarities, especially with respect to the lactiferous ducts and lymphatic drainage. In contrast, female rats have only one lactiferous duct per nipple. A comprehensive comparison between humans and dogs is given a special focus, as these species share several aspects in terms of breast/mammary cancer epidemiology, such as age of onset, hormonal etiology, risk factors, and the clinical course of the disease. Holistically, it is clear that each species has advantages and limitations that researchers must consider during the development of experimental designs and data analysis. [ABSTRACT FROM AUTHOR]

Abstract Background Mammary gland tumours are the most frequently diagnosed tumours in the female dogs but just a few studies have analysed their epidemiology. Therefore, we set out to describe the epidemiology of canine mammary cancer in the Canary Archipelago, Spain. We analysed a pathology tumour registry (PTR) and identified 7362 samples obtained from 5240 female dogs resident on the Canary Archipelago during an 18-year period (2003–2020). Using a case–control study design, we compared mammary tumour affected dogs with the Canarian canine population registry in order to elucidate the breed associations for these tumours. Results The frequency of a diagnosis of mammary tumours relative to all tumour diagnoses in female dogs decreased during the study period from 62.7% to 48.9%. Contemporaneously, the proportion of dogs diagnosed with mammary tumours who were also neutered increased from 13.6% to 26.9%. There was a negative correlation (R = -0.84) between these changes. Additional findings were that: the proportion of female dogs diagnosed with multiple tumours increased by 23.5% and that the proportion of malignant tumours 89.2% diagnosed has remained stable through the period. Benign mammary tumours were diagnosed at younger ages (9.2 years old) than carcinomas (9.7 years old) and sarcomas (10.4 years old). Epithelial mammary tumours were diagnosed at younger ages in entire female dogs. Samoyed, Schnauzer, Poodle, German Pinscher and Cocker Spaniel were the breeds with the highest odds-ratios (OR) in comparison with the reference (crossbreeds) while Miniature Pinscher, American Staffordshire Terrier, English Pointer as well as some local breeds such as the Canary Warren Hound and the Majorero had the lowest ORs. Conclusions This study provides a description of the changing epidemiology of canine mammary cancer in the Canary Archipelago over the last two decades. We found high rates of CMT with a significant predominance of malignant tumours. Exact risk factors are uncertain, but a combination of environmental, regional socioeconomic affecting human and their pets, and animal management factors are likely to play a part. Specifically, neutering was negatively associated with the proportion of epithelial mammary gland tumours and breeds native to the region were at lower risk of mammary tumours. A deeper analysis of all these factors will facilitate a deeper understanding of the epidemiology of mammary gland tumours in both the canine and the human population.

Background: Hematological analyses are seen as more preferred laboratory analyses in canine transmissible venereal tumor studies. There is no information about the availability of platelets and their indices in routine practice in canine transmissible venereal tumor cases. Taking this as a starting point, this study analyzed the usefulness of platelet indices in dogs with transmissible venereal tumor in clinical laboratory diagnosis as well as examined the relationship between white blood cells, red blood cells, platelets (PLT), main platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), and the ratio of main platelet volume to platelets (MPV/PLT). Materials, Methods & Results: In the study, a total of 42 bitches of various breeds were used. Nineteen healthy bitches were used as a control group, and the others 23 with cTVT as a study group. Metastasis was not observed in any of the bitches involved in the study. History, clinical findings, and cytological examinations were evaluated for the diagnosis of cTVT. In animals with hemorrhagic discharge and neoplastic lesions, a vaginal cytological examination was performed. Typical TVT cells with large nuclei and intracytoplasmic vacuoles were observed in the vaginal cytological examinations, and the diagnosis of TVT was made. Healthy bitches (19) and those with TVT (23) were 39.16 5.37 months and 47.61 5.14 months old, respectively. From all animals, 2 mL blood samples were collected from V. cephalica to evaluate PIs in the complete blood count (CBC). Collected blood samples were analyzed using an automated hematology analyzer. As a result of the analysis, WBC, RBC, HGB, HCT, MCV, MCHC, RDW, PLT, MPV, PDW, PCT, and MPV/PLT data were obtained. Mild leukocytosis, an increase in PLT, and a decrease in MCV and MPV/PLT were determined in the study group compared to the control group. Cut-off values in CBC of bitches with TVT were determined as WBC: 13.35 (sensitivity: 78%; specificity: 90%); MCV: 67 (sensitivity: 57%; specificity: 95%); PLT: 315.50 (sensitivity: 65%; specificity: 74%); and MPV/PLT: 0.028 (sensitivity: 78%; specificity: 58%). In CBC analyses, a strong negative correlation between PLT and MPV/PLT was detected in both groups. Discussion: Canine transmissible venereal tumors are common in both stray and pet dogs. It is naturally transferred from animal to animal during mating by live tumor cells. This tumor can commonly affect the external genitalia and internal organs in some cases. It generally has the look of cauliflower, and its surface is ulcerated, inflammatory, hemorrhagic, and infectious. More preferred laboratory analyses are complete blood count and blood chemistry analysis in cTVT for to evaluate the success of treatments. Platelet indices have been investigated in many diseases such as endotoxemia, chronic enteropathy, mammary tumor, parvoviral enteritis, septic peritonitis, lymphoma, pyometra, visceral leishmaniasis, and babesiosis in dogs. There is no information available for either diagnostic or prognostic use of the PIs in canine TVT cases. Ultimately, in light of the presented study's results, platelet indices, especially PLT and the MPV/PLT ratio, seem to be notable laboratory markers in terms of easy accessibility and low-cost assessment techniques in canine transmissible venereal tumor cases. New data, however, should be established by a thorough follow-up study using a larger sample size and addressing its usefulness as a diagnostic or prognostic marker in canine transmissible venereal tumors. [ABSTRACT FROM AUTHOR]

Background: Mammary gland tumours are the most frequently diagnosed tumours in the female dogs but just a few studies have analysed their epidemiology. Therefore, we set out to describe the epidemiology of canine mammary cancer in the Canary Archipelago, Spain. We analysed a pathology tumour registry (PTR) and identified 7362 samples obtained from 5240 female dogs resident on the Canary Archipelago during an 18-year period (2003–2020). Using a case–control study design, we compared mammary tumour affected dogs with the Canarian canine population registry in order to elucidate the breed associations for these tumours. Results: The frequency of a diagnosis of mammary tumours relative to all tumour diagnoses in female dogs decreased during the study period from 62.7% to 48.9%. Contemporaneously, the proportion of dogs diagnosed with mammary tumours who were also neutered increased from 13.6% to 26.9%. There was a negative correlation (R = -0.84) between these changes. Additional findings were that: the proportion of female dogs diagnosed with multiple tumours increased by 23.5% and that the proportion of malignant tumours 89.2% diagnosed has remained stable through the period. Benign mammary tumours were diagnosed at younger ages (9.2 years old) than carcinomas (9.7 years old) and sarcomas (10.4 years old). Epithelial mammary tumours were diagnosed at younger ages in entire female dogs. Samoyed, Schnauzer, Poodle, German Pinscher and Cocker Spaniel were the breeds with the highest odds-ratios (OR) in comparison with the reference (crossbreeds) while Miniature Pinscher, American Staffordshire Terrier, English Pointer as well as some local breeds such as the Canary Warren Hound and the Majorero had the lowest ORs. Conclusions: This study provides a description of the changing epidemiology of canine mammary cancer in the Canary Archipelago over the last two decades. We found high rates of CMT with a significant predominance of malignant tumours. Exact risk factors are uncertain, but a combination of environmental, regional socioeconomic affecting human and their pets, and animal management factors are likely to play a part. Specifically, neutering was negatively associated with the proportion of epithelial mammary gland tumours and breeds native to the region were at lower risk of mammary tumours. A deeper analysis of all these factors will facilitate a deeper understanding of the epidemiology of mammary gland tumours in both the canine and the human population. [ABSTRACT FROM AUTHOR]

Breast cancer is one of the most common and well-known types of cancer among women worldwide and is the most frequent neoplasm in intact female dogs. Female dogs are considered attractive models or studying spontaneous breast cancer, whereas female rats are currently the most widely used animal models for breast cancer research in the laboratory context. Both female dogs and female rats have contributed to the advancement of scientific knowledge in this field, and, in a “One Health” approach, they have allowed broad understanding of specific biopathological pathways, influence of environmental factors and screening/discovery of candidate therapies. This review aims to clearly showcase the similarities and differences among woman, female dog and female rat concerning to anatomical, physiological and histological features of the mammary gland and breast/mammary cancer epidemiology, in order to better portray breast tumorigenesis, and to ensure appropriate conclusions and extrapolation of results among species. We also discuss the major aspects that stand out in these species. The mammary glands of female dogs and women share structural similarities, especially with respect to the lactiferous ducts and lymphatic drainage. In contrast, female rats have only one lactiferous duct per nipple. A comprehensive comparison between humans and dogs is given a special focus, as these species share several aspects in terms of breast/mammary cancer epidemiology, such as age of onset, hormonal etiology, risk factors, and the clinical course of the disease. Holistically, it is clear that each species has advantages and limitations that researchers must consider during the development of experimental designs and data analysis.

Uterine prolapse is a rare condition in female dogs, defined by the eversion and protrusion of a portion of the uterus through the cervix into the vagina during or days after delivery, associated with excessive force and is considered an emergency. Treatment takes place by assessing the viability of the prolapsed uterus. Thus, this study aims to describe a case of uterine prolapse in a female dog, identified in the city of Imperatriz - MA, as well as the therapeutic-surgical approach adopted and its proficiency, since there is little information reported about this condition, where immediate care of uterine prolapse proved to be essential for a favorable prognosis, and the realization of ovariohysterectomy (OVH) presented itself as an efficient method for treatment. [ABSTRACT FROM AUTHOR]

Background: Aberrant right subclavian artery is only rarely observed in veterinary medicine. Some animals may present postprandial regurgitation and progressive weight loss, which is considered an incidental finding unrelated to clinical alterations. Advanced imaging techniques such as thoracic CT scan, magnetic resonance imaging (MRI) and contrast angiography are used for the accurate detection of lesions, anatomical changes and specific information about vascular rings. This paper describes the clinical changes, imaging exams and therapeutic approach in a female dog with megaesophagus induced by an aberrant right subclavian artery. Case: A 2-month-old female bull terrier, weighing 1.6 kg, with a history of regurgitation immediately or a few minutes after a meal, diarrhea, polyphagia, progressive emaciation and apathy for 45 days, was treated at a University Veterinary Hospital. The dog’s physical examination revealed 7% dehydration and body condition score 1 (scale 1 to 5), but no cardiac or pulmonary alterations upon auscultation. The hematological analysis and renal and hepatic serum enzymes were within the normal range for the species. In view of the presumptive clinical diagnosis of vascular anomaly, suggested by the contrast X-ray examination, a chest tomography was performed, which revealed altered aortic arch shape and contours, and a posterior aneurysm in the area of abnormal connection of the right subclavian artery. The patient was released with a prescription for conservative dietary management for megaesophagus. Within two weeks, the patient returned with a report of a good response to the prescribed therapy, absence of vomiting and diarrhea, and an increase in body weight. Surgical correction was recommended, but has not been performed so far, but conservative treatment for megaesophagus was continued. No further episodes of regurgitation were identified during the nine-month follow-up period. Discussion: In the case reported here, the right subclavian artery is considered anatomically atypical because it arises directly from the aortic arch. This vascular anomaly passes on to the right pectoral limb, dorsal to the esophagus, contracting it in its dorsal aspect. It tends to affect purebred dogs, occurring more frequently in Irish setters, German shepherds and Labrador retrievers, although it has been described in other breeds such as the bull terrier documented here and mixed breed dogs. Vascular ring anomalies may not cause clinical changes in animals and represent only incidental findings, or they may lead to gastrointestinal changes resulting from esophageal stricture, contributing to megaesophagus and clinical signs of esophageal obstruction, especially in recently weaned puppies. Such alterations were observed in this case, with the dog presenting regurgitation, immediately or a few minutes after a meal, megaesophagus, diarrhea and progressive weight loss. A CT scan was performed to confirm the type and location of the vascular anomaly and diagnostic accuracy, as recommended in the literature. Dietary therapy is one of the approaches adopted for patients presenting with regurgitation resulting from megaesophagus secondary to vascular anomalies. The dog in this report responded well to the medical therapy; nevertheless, the treatment of choice to correct the esophageal obstruction caused by this anomaly is surgical sectioning of the aberrant vessel by right intercostal thoracotomy, given that the degree of esophageal dilation and dysfunction tends to increase over time. However, at this time, the animal’s owner decided to suspend the recommended surgical procedure. [ABSTRACT FROM AUTHOR]

Background and Aim: Ovariohysterectomy (OHE) is a common procedure for sterilization of female dogs. However, knowledge of changes in pain stress, oxidative stress, and total antioxidant power status before, during, and after OHE is limited. The objective of this experiment was to study the effect of duration on pain stress, oxidative stress, and total antioxidant power status in female dogs undergoing OHE. Materials and Methods: Seven female dogs were sterilized using the OHE method. Pain scores, hematological changes, and biochemical markers were investigated during pre-operative, 3 h after starting OHE, and on days 3, 7, 10, and 14 of an experimental period. Data were analyzed using one-way analysis of variance. Results: At 3 days after OHE, pain score was higher than on days 7-14 of the experimental period; percentage of neutrophil, 3 h after starting OHE, was higher than during pre-operative and on days 3-14 of the experimental period; percentage of lymphocyte on days 10-14 was lower than during pre-operative, 3 h after starting OHE, and on days 3-7 of the experimental period; neutrophil/lymphocyte ratio, 3 h after starting OHE, was higher than during pre-operative and on days 3-14 of the experimental period; plasma malondialdehyde on day 3 was higher than during pre-operative, 3 h after starting OHE, and on days 3-14 of the experimental period; and total antioxidant power on day 14 was higher than during pre-operative, 3 h after starting OHE, and on days 3-10 of the experimental period, respectively. Conclusion: This experiment indicated that OHE caused pain stress, oxidative stress, and reduction of total antioxidant power in female dogs. Finally, female dogs needed antioxidant for 7 days after OHE.

Vaginal cytology is an important examination method in the context of gynecological disorders and cycle staging in the bitch. While collection and preparation of samples are easy, the evaluation appears to be challenging. Inconsistent definitions of cell attributes such as size, cornification and the appearance of the nucleus have been published. The aim of the project was to develop a tutorial for vaginal cell determination. To get a deeper insight into the use of cytology in practice, an online survey was distributed to veterinarians interested in small animal reproduction. Participants were asked to define eight cells and answer questions. The agreement of the 16 participants, working in eight different countries, determining the cells was poor (κ = 0.412). Eleven respondents stated that vaginal cytology has a low reliability. Nevertheless, 13 participants use this tool regularly. The tutorial was developed as a flowchart based on the survey results, scientific literature and own measurements. It guides the user systematically through the evaluation of specific cell characteristics. An evaluation of the results of five raters with difference experience levels led to a high agreement (κ = 0.858). Vaginal cytology is a useful diagnostic tool, but it seems helpful to standardize the determination of cell types.

This study evaluated the accuracy of mammary carcinoma diagnoses in female dogs through cytological exams (FNA) compared to histopathological diagnoses. The presence of neoplasia and the effectiveness of procedures at the Pathology Laboratory of the Veterinary Hospital of the FMVZ of Unesp Botucatu, were analyzed. Between 2015 and 2020, a total of 1100 mammary neoplasms were identified, of which 569 were mammary carcinomas. Fifty cytological samples were selected and analyzed to determine occurrence, age at presentation, and the most affected breeds, as well as to verify the obtained diagnoses. Mammary carcinoma constituted for 51.72% of the registered cases. A higher occurrence was observed in mixed-breed female dogs, at 40.42%, followed by Poodles at 17%. The most common age at diagnosis was 10 years, and in 65.55% of cases, the dogs had not been previously spayed. 9.31% of the animals had received contraceptives, while 14% had given birth and 14.58% had presented symptoms of pseudopregnancy at some point in their lives. In the test results, a 70% agreement between cytology and histology was observed, with a 30% disagreement between them. Statistically, a sensitivity of 79.32% and a specificity of 57.14% were reflected. Intact and older female dogs represent a significant risk of developing mammary carcinoma. Although the protocol for processing and interpreting cytological samples is well established, the results do not reach the level of excellence observed in previous studies., Competing Interests: Conflict of interests: PAMC, NSR, NFS and FCD – No conflict of interest.

Invasive mammary carcinomas with neuroendocrine differentiation are rare in women and were reported only once in female dogs. For the present study, ten cases of solid mammary carcinoma positive for chromogramin A in immunohistochemistry were selected. Histopathological characteristics of these tumors were described and immunohistochemical evaluation was performed with chromogranin A, synaptophysin, CD56, NSE, PGP 9.5, pancitokeratin, Ki67, estrogen receptor (ER), and progesterone receptor (PR). The average animal age was 13.2 years old and the average tumor size was 4.8 cm. In total, 70% of the neoplasms were classified as grade III and 30% as grade II by the Nottingham histological grade system. High mitotic index was observed with a mean of 27.5 mitoses in 10 high magnification fields. Only one case showed typical carcinoid tumor characteristics. In addition, vascular invasion was shown in 3 tumors. All carcinomas were positive for chromogran A, while only two cases were reactive to synaptophysin. For PGP 9.2, NSE and CD56, we observed positivity of 100, 90, and 70%, respectively, in the samples, being that no tumor was positive for all the neuroendocrine markers. All neoplasms showed ER and PR in at least 10% of neoplastic cells, while Ki67 varied from 29 to 95%, with mean mitotic index of 67%. Four of the ten animals died within 1 year of the tumor diagnosis. Neuroendocrine neoplasms occur in the canine mammary gland and are propably underdiagnosed. This is due to their non-specific morphological characteristics and the low use of neuroendocrine immunohistochemistric markers the diagnostic routine. More studies are necessary to determine the prognosis of this new histological type.

Urethral sphincter mechanism incompetence (USMI) is a common cause of urinary incontinence in dogs. Although estrogen is often prescribed for the medical therapy of USMI for spayed female dogs, they are known to have limited effectiveness and potential adverse effects. In castrated male dogs with USMI, testosterone reagents have been attempted besides estrogen. In this study, the effect of testosterone drugs, mainly methyltestosterone, on spayed female dogs with USMI was retrospectively evaluated. Ten spayed female dogs with USMI were included. Diagnosis of USMI was based on the results of the dogs' medical history, clinical signs, and no abnormalities in physical examinations, urinalysis, ultrasonography, X-ray imaging, and neurological examinations. Methyltestosterone was administered at doses of 0.32-1.27 mg/kg BW p.o. semel in die (sid.) to twice a week. Nine of the ten dogs had good or excellent responses 2 to 4 weeks after the start of treatment. The minimum effective dose was 0.32 mg/kg/day. Although no severe adverse symptoms occurred in any dog, a mild increase in alanine aminotransferase was temporally observed at doses of 1.0 and 1.1 mg/kg/day in the two dogs. After dose reduction or withdrawal, two of eight dogs had recurrence of urinary incontinence. Resumption of testosterone treatment clearly improved the symptoms in the two dogs. These results indicate that testosterone reagents might be an option for treating USMI in spayed female dogs as well. [ABSTRACT FROM AUTHOR]

Mammary tumors affect intact and elderly female dogs, and almost 50% of these cases are malignant. Cell culture offers a promising preclinical model to study this disease and creates the opportunity to deposit cell lines at a cell bank to allow greater assay reproducibility and more reliable validation of the results. Another important aspect is the possibility of establishing models and improving our understanding of tumor characteristics, such as vasculogenic mimicry. Because of the importance of cancer cell lines in preclinical models, the present study established and characterized primary cell lines from canine mammary gland tumors. Cell cultures were evaluated for morphology, phenotype, vasculogenic mimicry (VM), and tumorigenicity abilities. We collected 17 primary mammary carcinoma and three metastases and obtained satisfactory results from 10 samples. The cells were transplanted to a xenograft model. All cell lines exhibited a spindle-shaped or polygonal morphology and expressed concomitant pancytokeratin and cytokeratin 8/18. Four cell lines had vasculogenic mimicry ability in vitro, and two cell lines showed in vivo tumorigenicity and VM in the xenotransplanted tumor. Cellular characterization will help create a database to increase our knowledge of mammary carcinomas in dogs, including studies of tumor behavior and the identification of new therapeutic targets.

Background/Aim: Mammary neoplasms, like breast neoplasms in women, are one of the most common tumours in female dogs. Cancer-associated fibroblasts (CAFs) found in the tumour stroma play a role in angiogenesis and increase cell migration, contributing to tumour growth and progression, as well as metastasis. The aim of our work was to determine the level of periostin (POSTN) expression in CAFs in mammary tumours of female dogs. Materials and Methods: The research material consisted of 77 carcinomas and 24 adenomas of the mammary ridge in female dogs. Immunohistochemistry tests were performed using antibodies directed against the antigens POSTN, Ki-67, ERB-B2 receptor tyrosine kinase 2 (HER2), vimentin, and alpha smooth muscle actin (αSMA). Expression of POSTN at the mRNA level was determined using real-time polymerase chain reaction methods in 20 cases of mammary neoplasms. Results: Expression of POSTN in CAFs was observed in 92% of mammary cancer samples and in 25% of mammary adenoma samples in female dogs. A statistically significant increase in POSNT expression in CAFs was found in the carcinomas compared with mammary adenomas in female dogs. Expression of POSTN in CAFs in mammary carcinomas in female dogs positively correlated with the histological malignancy grade of tumours and the expression of Ki-67 proliferative antigen. Conclusion: Our results suggest a role of POSTN on the pathogenesis of mammary tumours in female dogs. Moreover, POSTN may prove to be a useful marker in the evaluation of cancerous stroma of mammary tumours in female dogs, and may have prognostic significance. [ABSTRACT FROM AUTHOR]

Background and Aim: Ovariohysterectomy (OHE) is a common procedure for sterilization of female dogs. However, knowledge of changes in pain stress, oxidative stress, and total antioxidant power status before, during, and after OHE is limited. The objective of this experiment was to study the effect of duration on pain stress, oxidative stress, and total antioxidant power status in female dogs undergoing OHE. Materials and Methods: Seven female dogs were sterilized using the OHE method. Pain scores, hematological changes, and biochemical markers were investigated during pre-operative, 3 h after starting OHE, and on days 3, 7, 10, and 14 of an experimental period. Data were analyzed using one-way analysis of variance. Results: At 3 days after OHE, pain score was higher than on days 7-14 of the experimental period; percentage of neutrophil, 3 h after starting OHE, was higher than during pre-operative and on days 3-14 of the experimental period; percentage of lymphocyte on days 10-14 was lower than during pre-operative, 3 h after starting OHE, and on days 3-7 of the experimental period; neutrophil/lymphocyte ratio, 3 h after starting OHE, was higher than during pre-operative and on days 3-14 of the experimental period; plasma malondialdehyde on day 3 was higher than during pre-operative, 3 h after starting OHE, and on days 3-14 of the experimental period; and total antioxidant power on day 14 was higher than during pre-operative, 3 h after starting OHE, and on days 3-10 of the experimental period, respectively. Conclusion: This experiment indicated that OHE caused pain stress, oxidative stress, and reduction of total antioxidant power in female dogs. Finally, female dogs needed antioxidant for 7 days after OHE. [ABSTRACT FROM AUTHOR]

Background/Aim: Mammary neoplasms are very common tumours in female dogs. Cancer-associated fibroblasts (CAFs) play an important role in the oncogenesis process. One of the useful proteins used in the diagnostics of CAFs cells is podoplanin (PDPN). The aim of our study was to assess the expression of PDPN in mammary cancer in female dogs. Materials and Methods: Our study cohort included 61 cancers and 21 adenomas of the mammary tumour in bitches. Expression of podoplanin, Ki-67 and HER2 was determined using the Immunohistochemical (IHC) method. PDPN expression at the mRNA level was determined using real-time PCR. Results: Expression of PDPN in CAFs was observed in 22.9% of cases of mammary cancers in bitches, with no PDPN expression in adenomas. A positive correlation was found between the expression of PDPN in CAFs and the grade of histological malignancy and expression of Ki-67. Conclusion: PDPN plays a significant role during the process of carcinogenesis of mammary tumours in female dogs. [ABSTRACT FROM AUTHOR]

The canine mammary tumor is the neoplasia that most commonly affects intact female dogs (not spayed) in routine veterinary practice. Canine mammary tumor therapy is a challenge because only few effective treatments have been described for high-grade tumors in veterinary medicine. One such therapeutic option that slows down tumor growth is metronomic chemotherapy (MC), a therapeutic modality that acts by decreasing tumor angiogenesis. Quantification of intratumoral microvessel density (MVD) has been proposed as a means to evaluate the effectiveness of MC in human and canine tumors. In this study, MVD is proposed as a predictive factor for the effectiveness of MC in canine mammary tumors. Twenty female dogs with mammary carcinoma were equally distributed into a mastectomy group treated only with mastectomy, and a MC group treated with MC (cyclophosphamide and piroxicam) orally and daily for 28 days followed by mastectomy. Mammary tumors were classified and graded histologically. MVD was ascertained by CD31 immunostaining. The analysis showed statistically significant difference in MVD scores between groups, corroborating a quantitative reduction in tumor microvasculature in the group treated with MC. Our findings suggest that MVD may be an important predictive factor for the selection of female dogs with malignant mammary tumors that may benefit from MC.

Rapamycin is an antifungal drug with antitumor activity and acts inhibiting the mTOR complex. Due to drug antitumor potential, the aim of this study was to evaluate its effect on a preclinical model of primary mammary gland tumors and their metastases from female dogs. Four cell lines from our cell bank, two from primary canine mammary tumors (UNESP-CM1, UNESP-CM60) and two metastases (UNESP-MM1, and UNESP-MM4) were cultured in vitro and investigated for rapamycin IC50. Then, cell lines were treated with rapamycin IC50 dose and mRNA and protein were extracted in treated and non-treated cells to perform AKT, mTOR, PTEN and 4EBP1 gene expression and global proteomics by mass spectrometry. MTT assay demonstrated rapamycin IC50 dose for all different tumor cells between 2 and 10 μM. RT-qPCR from cultured cells, control versus treated group and primary tumor cells versus metastatic tumor cells, did not shown statistical differences. In proteomics were found 273 proteins in all groups, and after data normalization 49 and 92 proteins were used for statistical analysis for comparisons between control versus rapamycin treatment groups, and metastasis versus primary tumor versus metastasis rapamycin versus primary tumor rapamycin, respectively. Considering the two statistical analysis, four proteins, phosphoglycerate mutase, malate dehydrogenase, l-lactate dehydrogenase and nucleolin were found in decreased abundance in the rapamycin group and they are related with cellular metabolic processes and enhanced tumor malignant behavior. Two proteins, dihydrolipoamide dehydrogenase and superoxide dismutase, also related with metabolic processes, were found in higher abundance in rapamycin group and are associated with apoptosis. The results suggested that rapamycin was able to inhibit cell growth of mammary gland tumor and metastatic tumors cells in vitro, however, concentrations needed to reach the IC50 were higher when compared to other studies.

Mammary hyperplasia is characterized by an increase in the whole mammary gland, considered benign and with a good prognosis. A two-year-old dog showed a volume increase in the entire mammary chain after hormone stimulation. To characterize the process prognosis, a immunohistochemical procedure was used to evaluate the Maspin staining, considered good prognosis marker, and the Ki-67, a proliferation marker, whose high expression indicates malignancy. As a result, we obtained a low expression of Ki-67 and a high expression of Maspin, suggesting an excellent prognosis for the animal.

Mammary gland tumours occupy a significant place in the pathology of this species. Female dogs that are not spayed after their first heat cycle have a higher predisposition of developing mammary tumours. Most tumours metastasize at distance via the lymphatic system. In these conditions, the sentinel lymph nodes of the mammary glands must be assessed prior to surgical treatment. Considering the insufficient usage of non-invasive investigative methods of the sentinel lymph nodes, the aim of this study is to describe the sonographic anatomy of the lymph nodes that drain the mammary gland tumours in female dog. Twelve dog females presenting tumours of the cranial and caudal abdominal mammary glands (A1 and A2), inguinal mammary gland (I) and cranial thoracic mammary gland (T1) were examined (group I). In addition, a control group composed of eight dog females was used (group II). The axillary and superficial inguinal lymph nodes were evaluated using an algorithm composed of gray-scale ultrasound, Doppler technique, contrast enhanced ultrasound (CEUS) and real time elastography. Surgical excision of the sentinel lymph nodes was performed and samples for histopathological examination were taken. The following ultrasonographic findings revealed on gray-scale examination were suspected for the metastatic infiltration: hypoechoic pattern, round shape, hillus absence and heterogenicity. Doppler technique showed an aberrant and mixed vascularisation of the lymph nodes, while the CEUS revealed incomplete enhancement of lymph nodes parenchyma. On real time elastography, the presence of blue areas in more than 50% from the lymph nodes parenchyma led us to conclude that the lymph node stiffness was caused by metastatic infiltration. Histopathological examination confirmed the presence of the metastatic infiltration in 97% of the examined lymph nodes. The algorithm composed of gray-scale ultrasound, Doppler technique, CEUS and real time elastography proved to be efficient in diagnosing the metastatic infiltration of sentinel lymph nodes of mammary gland. Further studies are needed to validate the proposed algorithm. [ABSTRACT FROM AUTHOR]

The objective of this work was to determine fertilization rates and embryonal recovery and to evaluate aspects of the early embryonal development. Eleven adult female dogs were used, with no defined breed. After the beginning of the natural estrus, these were mated and subjected to ovariohisterectomy on the 5th, 7th or 9th day postcoitus. Ovaries, tubes and uterine horns were separated. From the ovaries, the number of corpora lutea was determined. Tubes and uterine horns were washed separately in order to collect ovules and embryos, which were identified through the use of a stereomicroscope at 15x magnification. The fertilization rate was 94% and embryonal recovery 68.5%. On the fifth day, postcoitus embryos of up to eight cells were found in uterine tubes. Morulae or blastocysts were observed in uterine horns from the 9th day after recovery. On the basis of these results, it was concluded that the loss of structures can be due to the failure of their uptake by uterine tubes or during the execution of the technique. A high fertilization rate can be achieved even with the use of a single covering. It was also concluded that the embryonal development in uterine tubes had a duration of up to five days postcoitus with a stage of up to eight cells, embryos being found in stages of morulae in uterine horns nine days postcoitus.

The article discusses the efficacy of intratumoural injection of NS1 gene of canine parvo virus in mammary gland tumours (MGT) of female dogs. Topics discussed include the physical and clinical methods of observing the tumours pre and post treatment, and histopatological and electromicroscopic studies of the biopsy samples of tumuor tissues. Results of the study suggests that NS1 gene of canine parvovirus have apoptopic activity in regressing MGT.