The serum concentration of certain acute phase proteins significantly increases during various pathological conditions in cattle. The aim of this study was to determine the influence of claw disorders etiology on the concentrations of two major acute phase proteins in dairy cattle: haptoglobin (Hp) and serum amyloid protein A (SAA). Fifty dairy cows with claw pathology were included. Fourteen clinically healthy heifers served as controls. The animals were subdivided in 5 groups according to the pathological findings on their claws: 1. Heel horn erosion (HE), 2. Acute laminitis (AL), 3. Sole ulcer (SU), 4. Digital dermatitis (DD) and 5. White line separation (WLS). Hp and SAA concentrations were measured in serum samples using commercial ELISA kits. Higher concentrations of both Hp and SAA were found in the AL and SU groups (p<0.01) compared to the HE, DD and WLS and control groups. Dairy cows in the DD group had higher (p<0.05) Hp and SAA concentrations than the HE and WLS groups and the controls. The serum values between the HE, WLS and the control group did not differ significantly. The presented results indicate that the claw diseases are associated with a systemic acute phase response. Hp and SAA could be used as valuable biomarkers for early detection of claw diseases in dairy cows.
Postpartum anestrus is a physiological phenomenon in high-producing dairy cows. Static ovaries have been related as major contributors for its occurrence causing a significant reproductive problem to the dairy industry. Different treatment methods have been employed with inconsistent rate of success in initiation of cyclicity, requiring further investigations in order to achieve satisfactory results. The aim of the present study was to compare the ovarian response in cows diagnosed with static ovaries, more than 60 days postpartum using two different hormonal treatment (GnRH and eCG) methods. A total of 58 acyclic cows (no CL, follicles<8mm, P4<0.5ng/mL) were randomly divided into three groups: GnRH (Group 1, n=23), eCG (Group 2, n=23) and Controls (n=12), and allocated thereafter, into subgroups according to the applied doses of GnRH (100μg or 250μg); eCG (750 IU or 1000 IU) whilst control group cows were left untreated. Daily follicular growth rate and treatment respond interval were estimated based on repeated ultrasound examinations. Blood serum P4 sampling was done on d -7, d-0 (start of the experiment) and on d 7 after ovulation. Resumption of cyclic activity occurred in 55.17% (32/58) of the treated cows, 56.52% in Group 1; 60.86% in Group 2 and 41.66% in the control group. Overall, the follicular growth rate was similar between the trials group and significant with regard to the cows in the control group (p<0.05). eCG or GnRH treated cows responded significantly faster 6.85±0.2 and 7.84±0.2 days, respectively, in comparison to the control group cows (17±0.7 days, p<0.001). Treatment with a single dose of GnRH or eCG caused resumption of follicular growth and ovulation following luteogenesis (forming CL) without significant changes in P4 concentrations on day 7 after ovulation (p>0.05). Cows in Group 2 had significantly higher incidence of multiple ovulations than cows in Group 1 (p<0.05). The eCG treatment resulted in a faster response and higher ovulation rate compared to GnRH treatment. In conclusion, both treatments have shown acceptable results in resumption of cyclicity in dairy cows with static ovaries.
The bovine kappa-casein (κ-CN) is a phospho-protein with 169 amino acids encoded by the CSN3 gene. The two most common gene variants in the HF breed are CSN3*A and CSN3*B while CSN3*E has been found with lower frequency. The aim of this study was to optimize a laboratory method for genotyping of these three alleles as well as to determine their genotype and allele frequencies in the HF cattle population in the Republic of North Macedonia. Genomic DNA was extracted from full blood from 250 cows. The target DNA sequence was amplified with newly designed pair of primers and the products were subjected to enzymatic restriction with HindIII and HaeIII endonucleases. Genotype determination was achieved in all animals. The primers successfully amplified a fragment of 458 bp and the digestion of this fragment with both endonucleases enabled differentiation of five different genotypes with the following observed frequencies: AA (0.39), AB (0.29), BB (0.16), AE (0.10), and BE (0.06). The estimated allele frequencies were: CSN3*A (0.584), CSN3*B (0.336) and CSN3*E (0.08). The observed genotype frequencies differed significantly (P<0.01) from those that would be expected under HW equilibrium, while the fixation index (F=0.17) indicated moderate heterozygosity deficiency. Nevertheless, the CSN3*B allele was present with relatively high frequency which should be used to positively select for its carriers, since increasing its frequency could help to improve the rheological properties of the milk intended for cheese production.
Mammary tumours are the second most frequent neoplasia in dogs, mainly affecting older female patients. Approximately 50% of the mammary tumours are malignant with high percentage of mortality, if not treated in time. The aim of this study was to analyze the data of canine patients with mammary tumours, to evaluate the type of tumours, as well as the relationship between tumour incidence and dogs’ age, reproductive cycle and sterilization. The survey was used to retrieve the information in the period of two years from the patient data base of the University Veterinary Hospital at the Faculty of veterinary medicine in Skopje. Patients included in this survey were subjected to routine clinical investigation and additional laboratory tests (cytological examination, x-rays imaging, CBC and biochemical profile, histopathology of the tumor samples). Aged female patients (12 - 13 years) are the most susceptible category for development of mammary tumours. The reproductive history showed that five of the patients with malignant mammary tumours have never whelped and were not treated with any exogenous hormones. Malignant tumours (adenocarcinoma) were diagnosed in 90% of the patients. Three patients died due to lung metastasis. Late diagnosis is one of the major problems that results in lethal outcome due to lung metastases. Since ovarian steroids play an important role in the aetiology, the most effective prevention of mammary tumours is elective ovariectomy of the bitch at an early age.
Ehrlichia canis, the etiologic agent of canine monocytic ehrlichiosis (CME), is mainly transmitted by the brown dog tick Rhipicephalus sanguineus. Clinical signs of the disease can be various, depending on the stage of the disease. Typical changes in hematological and blood biochemical parameters are: severe thrombocytopenia, mild to marked non regenerative anaemia and hypoalbuminemia. In order to present the effects of the treatment protocol on several hematological and biochemistry parameters, 34 Ehrlichia canis positive dogs were compared before and after treatment with doxycycline 10mg/kg/day, in duration of four weeks. Besides the clinical sings and laboratory findings, diagnosis was confirmed by antibody tests (Bionote, Korea, AGROLABO S.p.A., Italy). The most common clinical sings were depression, lethargy, pyrexia, vomiting and anorexia. Hematological analyses were performed on the automatic hematology analyzer Exigo EosVet (Sweden), while biochemistry analyses (alanine aminotransferase, aspartate aminotransferase, urea, creatinine, albumin, total protein, globulin and alkaline phosphatase) were performed using the automatic analyzer ChemWell 2910 (Awareness Technology, INC, USA). Statistically significant difference (p<0.05) in hematology changes was present regarding the red blood cells count, platelet count, hematocrit and hemoglobin before and after treatment. Hypoalbuminaemia (Mean 19.21 ±4.96 g/l) was the only serum biochemistry parameter with significant change before and after treatment, as well. Treatment with doxycycline in patients with E.canis resulted in significant increase of hematology parameters (red blood cells, hemoglobin, haematocrit and platelets), as well as albumins in serum.
Reduced glutathione (GSH) and homologous ram seminal plasma (HSP), used as additives in cryopreserving (CP) media prior to freezing, showed conflicting results in retaining structural integrity and progressive motility in post-thawed ram spermatozoa. The aims of this research were: (1) to assess the effect of GSH and/or HSP supplementation via soybean-lecithin CP extender on cryopreserved ram spermatozoa viability, morphology and motility pattern; and (2) to assess the effect of incubation in the context of the previous aim. Quantitatively and qualitatively, homogenized and pooled ram ejaculates (N=10) were extended with one of the following extenders: control (C) – tris-based, GSH and HSP-free, experimental-1 (E1) – C + GSH 5 mM, experimental-2 (E2) – C + HSP 20 % and experimental-3 (E3) - GSH 5 mM + HSP 20 %. Following thawing, samples were taken at 0- and 3-hours from each group (n=10) and were assessed for spermatozoa viability, morphology, and motility pattern. C-0h samples yielded a spermatozoa population with low viability, altered head morphology and highly deviated motility pattern. E3-3h samples yielded spermatozoa with unaffected viability, head morphology and high progressive motility. In conclusion, E3 extender added to cryopreserved-thawed ram spermatozoa is most efficient in obtaining high viability, unaltered head morphology, and progressive motility.
The present study aimed to evaluate whether the induction and the formation of an accessory corpus luteum (CL) after AI might increase the pregnancy per AI (P/AI) in heat stressed dairy cows. Starting at d 50±3 post-partum, 113 lactating Holstein cows from one commercial herd during summer were scored for body condition, blood sampled and examined by ultrasound. Those bearing a CL>25mm and progesterone (P4) level>2ng/mL were synchronized using a double PGF2α injection given 12 h apart and AI-ed at detected estrus. In total 18 cows, there were not any signs of estrus (n=10) nor a P4 level <2ng/mL at the time of enrolment (n=8) and therefore they were excluded from the study, leading to 95 cows finally enrolled.. At d5 post-AI, cows were randomly allocated into 2 groups: control group (CON, n=45) without any additional treatment, and treatment group (GnRH, n=50), treated with 0.008 mg Buserelin – a GnRH agonist. Blood sampling and ultrasound examination were done at d5, d14 and at d21 after AI, whereas the pregnancy diagnosis was done at d21 and d30 after AI. Average daily temperature and relative humidity values were used to calculate the temperature-humidity index (THI). The average THI during the experiment was 79.5±0.6. At d5, no differences were observed neither between the number of the CL nor between the P4 level in both groups. At d14 and d21, 82% of the GnRH-treated cows had more than one CL versus 0% of the CON cows. Both at d14 and 21, GnRH-treated cows had higher P4 levels compared to the CON cows (p<0.05). In addition, P/AI were higher in the GnRH group than in the CON group (65% vs. 48.3%, p<0.05), whereas late embryonic losses were higher in CON in comparison to GnRH cows (10.6 vs. 4%, respectively). The BCS at the moment of insemination did not affect P/AI (p>0.05). In conclusion, the induction of an accessory CL at d 5 after AI might increase P/AI in heat stressed dairy cows.
We hypothesized that a single dose of PGF2α belatedly injected on day 8 after GnRH-1 in cows receiving a 7-day Ovsynch-56 protocol (GnRH – 7 days – PGF2α – 56h – GnRH – 16h – timed AI) will increase the proportion of cows with complete luteolysis. At day 35±3 postpartum, 70 lactating Holstein cows from one herd were scored for body condition and pre-synchronized with PGF2α and GnRH (3 days apart) and 7 days later submitted to an Ovsynch-56 protocol for first AI after random assignment to two treatments: (1) OV-7 (n=35) with an injection of PGF2α either on day 7; or (2) OV-8 (n=35) on day 8 after G1, respectively. Blood was collected before the first PGF2α, at day 7 and day 8 in OV-7 and OV-8, respectively, at AI and at 7 days after AI to assess progesterone concentration. Ten cows were classified as acyclic and were excluded from the analysis resulting in 60 cows (OV-8, n=27; OV-7, n=33). In total, more (P=0.01) OV-8 cows and more (P=0.04) primiparous OV-8 cows had complete luteolysis compared with their OV-7 herd mates. In addition, more (P=0.008) OV-8 cows with BCS<2.75 had complete luteolysis compared with their OV-7 herd mates, whereas no difference was observed between treatments among cows with BCS ≥2.75. In conclusion, delaying the application of PGF2α by 1 day reduced the percentage of primiparous cows and cows with poorer BCS having incomplete luteal regression at the time of AI.
Mammary tumors are among the most common neoplasms in intact female dogs. They have a complex morphology, usually affecting middle age and older bitches. Almost 50% of the mammary tumors in dogs are malignant neoplasms. Prognosis is based on several factors: stage, age, tumor size, metastasis, histopathology, ovariectomy status and hormone-receptor activity. Immunohistochemical (IHC) measurement has become increasingly an important diagnostic and prognostic parameter, with the development of monoclonal antibodies against nuclear estrogen and progestin receptors. The aim of this study was to detect the presence of ER receptors in malignant canine mammary tumors and to identify their association with the clinical course of the tumor. Mammary tumor samples have been obtained by mastectomy from dogs presented at our clinic. Detailed clinical examination, CBC and basic serum biochemical profile were performed in all patients. Surgery was the only treatment. Histopathological examination and immunohistochemical detection of estrogen α receptors (ERα) was performed on 8 formalin-fixed, paraffin-embedded tissue samples, using the PT LINK immunoperoxidase technique. Histopathological examination of the mammary tumor samples (n=11) revealed tubular adenocarcinoma (n=6,54.5%) and ductal adenocarcinoma (n=3, 27.3%), one patient with benign adenoma and one with mastitis. Patients with positive ER tumors are alive, without remission, while 3 of the patients that were ER negative died due to lung metastases. According to our results, it can be concluded that the appearance and development of canine mammary tumors is highly connected with ovarian steroid hormones and that immunostaining of the tumors may be used as a good prognostic parameter in these patients.