Diagnosis and treatment of mastitis in mares

Infection and inflammation of the udder (mastitis) is a common condition affecting all domestic mammals, but it appears to be less prevalent in mares than in dairy cows and dairy goats. The seemingly reduced incidence of mastitis in mares can be partially explained by the smaller size and relatively concealed location of the mare’s udder, coupled with a smaller storage capacity than cows and goats. Mastitis can affect lactating, peripartum, dry mares, mares at dry-off or prepubertal foals. Common clinical signs include swollen mammary tissue, abnormal mammary gland secretion, fever and anorexia; less common signs are hindlimb lameness and a swollen mammary vein. On rare occasions, mastitis pathogens can severely affect the nursing foal and mares may develop fibrotic tissue and consequent agalactia in the side(s) or quarter(s) affected. Based on the clinical presentation, mastitis can be classified as acute or chronic, and clinical or subclinical. Diagnosis is based on the clinical signs aided with aerobic culture and cytological evaluation of the gland secretion. In addition, these ancillary tests can also be used to assess prognosis and duration of treatment. Mares suffering from mastitis may present neutrophilia and hyperfibrinogenaemia. Treatment for mastitis includes antimicrobial therapy (systemic and/or locally), nonsteroidal anti-inflammatory drugs, frequent milking and cold hosing with/without hot-packing applied on the gland. While the frequent monitoring of mares after weaning and reducing food intake should be part of common practices at weaning, cleaning of the udder, control of insect populations and frequent milking of mares with a foal unable to nurse can also aid in preventing mastitis.     

Carcinoma of the mammary gland in a mare

A 20‐year‐old Quarter Horse mare was presented with a firm, nonpainful swelling near the axial margin of the left mammary gland. Ultrasound examination of the mass revealed a 35 mm poorly encapsulated, homogeneous mass within the parenchyma of the left mammary gland. Using histopathology and immunohistochemistry, the mass was diagnosed as a mammary carcinoma and showed positivity for cytokeratin 18 (CK18), vimentin and α‐smooth muscle actin. Additionally, the mRNA expression level of the oncogene cMyc did not show a significant upregulation, whereas p53, a well‐known tumour suppressor gene in breast cancer, was significantly reduced in comparison with healthy equine mammary gland tissue. To the authors' knowledge, this is the first report of a significant downregulation of p53 expression in a mammary carcinoma of a mare.     

Secretory carcinoma of the mammary gland in an 8-year-old Holstein-Friesian dairy cow

This study was conducted to examine the oxidative stress biomarkers in a cow diagnosed with a follicular cyst in her left ovary. Progesterone (P4) and plasma oxidative stress status was measured in 13 Holstein cows after synchronization of oestrus with controlled internal drug release (CIDR) and prostaglandinF2α (PGF2α) protocol. The presence and size of ovarian structures were monitored by transrectal ultrasound at 4 hourly intervals. Of the 13 cows, 12 were monitored until ovulation was detected and recorded, whereas one cow failed to ovulate and developed a follicular cyst. Oxidative stress biomarkers; reactive oxygen metabolites (ROMs), biological antioxidant potential (BAP), oxidative stress index (OSI), glutathione (GSH), ceruloplasmin and advanced oxidation protein products (AOPP) were measured in the cystic cow and compared to those of the 12 ovulated cows and are referred to as higher or lower if they are outside the mean ± standard error of mean of those of ovulated cows. The cystic cow had lower ROMs and OSI between 36 and 84 h after PGF2α injection and at 9 h, from 36 to 60 h after PGF2α injection respectively. On the other hand, antioxidant (BAP and GSH) was higher in the cystic cow compared to her ovulated herd mates. The observed imbalance between oxidant and antioxidant might have disrupted the physiological events for ovulation to occur, leading to cystic ovarian disease.     

Identification of novel tumour‐associated antigens in canine mammary gland tumour

Canine mammary gland tumour (MGT) is the most common neoplasm in female dogs and has similar biological characteristics to human MGT. Spontaneous canine MGT is a more attractive clinical model in oncological research than that of the murine experimental model. Tumour‐associated antigens (TAAs), which are produced in tumour cells, are applied as tumour markers, tumour vaccine antigens and molecular targets of therapeutic drugs. In this study, we have primarily identified 13 different TAAs of canine MGT by serological immunoscreening of cDNA expression library. The results of serological mini‐arrays of identified antigens showed that CCDC41 antigen specially reacted with 35% of sera from MGT‐dogs and did not react with control sera. We also found that HSPH1 mRNA expression levels increased significantly in MGT tissues. These findings will contribute to the development of diagnostic technologies and translational target therapies for dogs. Clinical relevance : HSPH1, which is strongly expressed in the tumour tissue, will be a possible vaccine antigen of canine MGT.     

Expression and significance of PTEN in canine mammary gland tumours

To explore the expression and clinical importance of the anti-oncogene phosphatase and tensin homologue deleted on chromosome 10 (PTEN) in canine mammary gland tumours, PTEN expression was compared in 50 cases of canine mammary tumour and four examples of normal mammary tissue using real-time quantitative PCR. PTEN expression was similar in benign mammary tumours and normal mammary tissues (P>0.05), but was lower in malignant tumours than in normal mammary tissues or benign mammary tumours (P<0.001). PTEN expression was also low in the lymph node metastases of malignant mammary tumours. The expression profile of PTEN in malignant mammary tumours compared to those without lymph node metastasis varied significantly. Low-level PETN expression might play an important role in carcinogenesis and the progression of canine mammary tumours, and PTEN protein detection might be useful in evaluating tumour development and prognosis.     

Malignant fibrous histiocytoma of the mammary gland in a mare

Mammary tumours are uncommon in mares. Sarcomas of the breast account for less than 1% of all human primary malignancies and, among these types of tumours, malignant fibrous histiocytoma (MFH) is very rare. This report describes a 6‐year‐old uniparous Paint mare that was presented for investigation of right mammary gland enlargement of 3 weeks' duration. The clinical and histopathological findings in this mare with malignant fibrous histiocytoma of the mammary gland as well as complications of bilateral mastectomy are detailed. At one year follow‐up there were no clinical signs of recurrence.     

Immunolocalization of lingual antimicrobial peptide (LAP) in the bovine mammary gland

Lingual antimicrobial peptide (LAP), a member of the β-defensin family in cows, is involved in the innate immune system and plays a crucial role in killing a large variety of microorganisms. The aim of the present study was to demonstrate the immunolocalization of LAP in the mammary glands of cows. A LAP antibody was raised in a rabbit by immunity with a synthetic 11 amino acid sequence out of a 42-amino acid sequence of the mature form of LAP. The specificity of the LAP antibody was checked using a competitive immunoassay and Western blotting. Paraffin sections of the mammary gland were immunostained with LAP antibody. In the competitive immunoassay, an increase of synthetic LAP concentration suppressed the optical density. Western blotting analysis for LAP revealed the presence of the LAP peptide in mammary alveolar tissue. When the mammary gland was immunostained with LAP antibody, epithelial cells of both infected and non-infected alveoli were immunopositive. These results indicate that LAP is localized in the epithelium of non-infected as well as infected alveolus in the mammary gland in cows.     

Photodynamic detection of canine mammary gland tumours after oral administration of 5‐aminolevulinic acid

5‐Aminolevulinic acid (5‐ALA) is widely used in photodynamic detection (PDD) and therapy. We evaluated the pharmacokinetics of 5‐ALA‐induced porphyrins and its effectiveness in PDD in dogs with mammary gland tumours (MGTs) following oral administration. Healthy dogs and those with MGTs (nine each) were orally administered 4 mg kg^?1 5‐ALA. Protoporphyrin IX (PpIX) was not detected in the plasma of healthy dogs but it peaked in dogs with MGT at 2 h after 5‐ALA administration. In the PDD study, 16 dogs with MGT were orally administered 40 mg kg^?1 5‐ALA, and MGT but not normal tissue showed red fluorescence after 2–4 h. Photon counts were 6635–63 890 and 59–4011 (median, 19 943 and 919) for MGT and non‐tumour tissues, respectively. Cell density strongly correlated with PpIX photon counts of MGT tissue of the dogs (R = 0.743, P = 0.0009). We suggest that 5‐ALA‐PDD might be an effective diagnostic tool for MGTs.     

奶牛乳腺的防御机制与乳腺炎病理学

乳头管的结构和内衬的角质蛋白是乳腺抵御微生物侵入的物理性屏障,角蛋白中的低级脂肪酸、阳离子蛋白质对侵入的病原微生物具有抗性;环境性致病菌是引起机体免疫应答的主要因子,巨噬细胞、单核细胞、中性粒细胞和淋巴细胞之间的相互协调组成了自然的免疫系统,它们通过分泌细胞因子、抗体和吞噬作用等活动来灭杀、清除侵入机体的病原体;乳中的可溶性因子具有免疫调节作用,对细菌的结构具有破坏性;妊娠期及围产期奶牛由于内分泌的变化而导致免疫抑制,结果造成临床型和亚临床型乳腺炎的发病率相对较高;牛奶体细胞计数对亚临床型乳腺炎的监控具有参考性。     

A review of equine sarcoid

Equine sarcoid is the most common tumour of horses and accounts for over half of all equine skin tumours. Six types of sarcoid based on gross appearance and clinical behaviour have been described including occult, verrucous, nodular, fibroblastic, mixed and malevolent. Common locations for sarcoid development include the periocular region, ear pinnae, lips, neck, extremities and ventrum (including groin region). Bovine papillomavirus (BPV) is causally associated with equine sarcoid with genetic haplotype, fly vectors and skin trauma identified as potential risk factors for development of the disease. Histopathology is required for definitive diagnosis of equine sarcoid but incomplete excision is thought to activate latent BPV and stimulate growth. Although there are no uniformly effective treatment options, several modalities have been successful in eliminating or managing equine sarcoid. Surgical excision, intratumoural chemotherapy, cryotherapy, hyperthermia, radiotherapy, immunotherapy and immune modulators are used with degrees of success relative to the accessibility and invasiveness of the tumour. Prevention of equine sarcoid may be facilitated by future development of vaccines against bovine papillomavirus.     

Determination of carcinoembryonic antigen and cancer antigen (CA 15‐3) in bitches with tumours on mammary gland: preliminary report

The aim of this work was to determine levels of carcinoembryonic antigen (CEA) and cancer antigen (CA 15‐3) in the blood serum of 45 bitches. A modified procedure was used to determine the CEA and CA 15‐3 markers with the human kits using the radioimmunoassay method. Samples collected from extirpated tumour of mammary glands were histologically processed and classified as per WHO guidelines. The average age of animals with tumour was 10.00 ± 2.2 years; for healthy bitches average age was 4.2 ± 3.2 years. Values of CEA and CA 15‐3 were considered positive, if they exceeded 0.23 ng mL^?1 and 7 IU mL^?1, respectively. Average levels of CEA in the tumour group were 0.25 ± 0.06 versus 0.20 ± 0.03 in healthy bitches (P = 0.0001). The average CA 15‐3 value in bitches with tumour was 8.58 ± 1.27 versus 5.14 ± 1.34 in healthy animals (P < 0.0001).     

Expression of the glutamine metabolism‐related proteins glutaminase 1 and glutamate dehydrogenase in canine mammary tumours

Glutamine metabolism is an important metabolic pathway for cancer cell survival, and there is a critical connection between tumour growth and glutamine metabolism. Because of their similarities, canine mammary carcinomas are useful for studying human breast cancer. Accordingly, we investigated the correlations between the expression of glutamine metabolism‐related proteins and the pathological features of canine mammary tumours. We performed immunohistochemical and western blot analysis of 39 mammary tumour tissues. In immunohistochemical analysis, the expression of glutaminase 1 (GLS1) in the epithelial region increased according to the histological grade (P < .005). In the stromal region, complex‐type tumours displayed significantly higher GLS1 intensity than simple‐type tumours. However, glutamate dehydrogenase expression did not show the same tendencies as GLS1. The western blot results were consistent with the immunohistochemical findings. These results suggest that the expression of GLS1 is correlates with clinicopathological factors in canine mammary tumours and shows a similar pattern to human breast cancer.     

大鼠不同时期乳腺组织及血液中催乳素变化规律

为探讨催乳素在大鼠不同时期乳腺及血液中的变化规律,选择雌性SD大鼠42只,分别为处女鼠、妊娠鼠(6 d、12 d、18 d)、泌乳鼠(6 d、12 d、18 d),每个时期6只.在指定的时间点处死大鼠取乳腺组织及血清.用放免法对催乳素表达的变化进行定量研究,用统计学方法进行处理,观察催乳素的变化规律.结果显示,血清PRL水平以处女期最高,妊娠开始急剧下降,随着妊娠进行又逐渐增加,分娩后随着泌乳进行期渐渐下降.处女期与其余各期差异显著(P＜0.05),整个妊娠期间以妊娠18 d为最高,且妊娠期间差异不显著(P＞0.05),泌乳6 d明显高于泌乳12 d、18 d(P＜0.05),泌乳12 d、18 d明显低于妊娠各期水平(P＜0.05).乳腺PRL变化模式与血清的变化不一致,处女期最高,随妊娠下降,泌乳期相对稳定,变化不大.处女期与其余各期差异显著(P＜0.05),妊娠6 d水平仅明显低于处女期(P＜0.05),而显著高于其余各期(P＜0.05).表明催乳素不仅参与了乳腺细胞的发育,而且在维持和启动泌乳方面也起到重要的作用.     

Invasive micropapillary carcinoma of the mammary glands in a mare

Liver cell turnover is very slow, especially compared to intestines and stomach epithelium and hair cells. Since the liver is the main detoxifying organ in the body, it does not come as a surprise that the liver has an unmatched regenerative capacity. After 70% partial hepatectomy, the liver size returns to normal in about two weeks due to replication of differentiated hepatocytes and cholangiocytes. Despite this, liver diseases are regularly encountered in the veterinary clinic. Dogs primarily present with parenchymal pathologies such as hepatitis. The estimated frequency of canine hepatitis depends on the investigated population and accounts for 1%–2% of our university clinic referral population, and up to 12% in a general population. In chronic and severe acute liver disease, the regenerative and replicative capacity of the hepatocytes and/or cholangiocytes falls short and the liver is not restored. In this situation, proliferation of hepatic stem cells or hepatic progenitor cells (HPCs), on histology called the ductular reaction, comes into play to replace the damaged hepatocytes or cholangiocytes. For unknown reasons the ductular reaction is often too little and too late, or differentiation into fully differentiated hepatocytes or cholangiocytes is hampered. In this way, HPCs fail to fully regenerate the liver. The presence and potential of HPCs does, however, provide great prospectives for their use in regenerative strategies. This review highlights the regulation of, and the interaction between, HPCs and other liver cell types and discusses potential regenerative medicine-oriented strategies in canine hepatitis, making use of (liver) stem cells.     

Gene expression and hormonal regulation of adiponectin and its receptors in bovine mammary gland and mammary epithelial cells

Although the functions of adiponectin, a differentiated adipocyte‐derived hormone, in regulating glucose and fatty acid metabolism are regulated by two subtypes of adiponectin receptors (AdipoRs; AdipoR1 and AdipoR2), those in ruminants remain unclear. Therefore we examined the messenger RNA (mRNA) expression levels of adiponectin and its receptors in various bovine tissues and mammary glands among different lactation stages, and the effects of lactogenic hormones (insulin, dexamethasone and prolactin) and growth hormone (GH) on mRNA expression of the AdipoRs in cultured bovine mammary epithelial cells (BMEC). AdipoRs mRNAs were widely expressed in various bovine tissues, but adiponectin mRNA expression was significantly higher in adipose tissue than in other tissues. In the mammary gland, although adiponectin mRNA expression was significantly decreased at lactation, AdipoR1 mRNA expression was significantly higher at peak lactation than at the dry‐off stage. In BMEC, lactogenic hormones and GH upregulated AdipoR2 mRNA expression but did not change that of AdipoR1. In conclusion, adiponectin and its receptor mRNA were expressed in various bovine tissues and the adiponectin mRNA level was decreased during lactation. These results suggest that adiponectin and its receptors ware changed in mammary glands by lactation and that AdipoRs mRNA expression was regulated by different pathways in BMEC.     

The relationship between tumour size and expression of prognostic markers in benign and malignant canine mammary tumours

Tumour size is considered one of the most important determinants of clinical staging in cancer patients. The aim of this study was to assess the value of tumour size as an indicator of the differentiation of mammary neoplasias in female dogs. The tumour, nodes metastates (TNM) system, based on primary lesion size, the extent of its dissemination to regional lymph nodes and the presence or absence of distant metastases, was applied to 120 female dogs diagnosed with mammary neoplasias. Paraffin blocks from 38 cases were selected and studied by immunohistochemical staining for prognostic and predictive markers of breast cancer. The Kaplan–Meier survival curve was estimated for 110 female dogs. Larger tumours (T3) were mostly malignant and showed lower expression of progesterone receptor and higher expression of cellular proliferation markers. Global survival time was shorter in female dogs with large tumour masses. This study highlights the importance of tumour size as a prognostic indicator of mammary neoplasias in female dogs.     

A review of equine mucocutaneous squamous cell carcinoma

Squamous cell carcinoma (SCC) accounts for approximately 20% of all equine mucocutaneous (MC) tumours and continues to present a therapeutic challenge to practitioners. Most MC‐SCC are locally invasive and slow to metastasise, but metastasis to local lymph nodes is not uncommon. The most common location for MC‐SCC is the periorbital region, with the eyelid most commonly affected. Although only 13% of MC‐SCC involves the external genitalia, MC‐SCC is the most common neoplasm of male genitalia. Equine caballus papillomavirus‐2 has recently been linked to MC‐SCC and may prove to be necessary for tumour development. Risk factors may include chronic exposure to ultraviolet light and chronic skin irritation. Horses developing genital MC‐SCC tend to be older compared to those with periorbital MC‐SCC. Histopathology is required for definitive diagnosis of MC‐SCC, although horse phenotype and lesion location may suggest MC‐SCC. Several treatment modalities have been successful in eliminating or managing MC‐SCC, with surgical excision and intratumoural chemotherapy yielding the best results. Other treatment options including cryotherapy, hyperthermia, radiotherapy and photodynamic therapy are often used as adjunctive therapies. Early recognition of tumours and prompt intervention are associated with a positive outcome.     

Haemoglobin in normal and neoplastic canine mammary glands

Four types of globins for oxygen transport are known in vertebrates, and the haemoglobin is responsible for carrying oxygen in blood. In this study, we found that haemoglobin was also expressed in canine mammary glands. Samples were taken from 26 malignant mammary tumors, 16 normal mammary glands and 10 other normal tissues. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting and mass spectrometry were used to investigate haemoglobin in mammary tissues. The results indicated that normal canine mammary glands expressed high levels of haemoglobin protein as shown by Coomassie blue staining. The identity of haemoglobin was confirmed by immunoblotting and mass spectrometry, and the mass spectrometry data revealed that both alpha-haemoglobin and beta-haemoglobin were expressed. Relative to normal mammary glands, the levels of haemoglobin expression in mammary tumors were lower. Our results also indicated that the haemoglobin was endogenously produced in mammary gland tissues and was not derived from the erythroid cells.