Oxytetracycline Persistence in Uterine Secretion after Intrauterine Administration in Cows with Metritis.

Puerperal metritis in cows is often treated with antibacterial drugs. The prudent use of antibiotics in farm animals requires state-of-the-art knowledge of their pharmacokinetics and data from sensitivity tests of pathogenic bacteria. Changes in oxytetracycline levels in the uterine secretion over time after intrauterine administration in cows with metritis were evaluated in relation to the sensitivity of pathogenic bacterial isolates. Oxytetracycline levels in plasma, milk and uterine secretion were determined via LC-MS/MS analysis. Pathogenic bacteria were isolated and their sensitivity to oxytetracycline was determined. The concentrations of oxytetracycline in the uterine secretion were 433.79 (39.17-1668.76) µg·mL-1 six hours after the third application at a dose of 8 mg/kg and 84.33 (1.58-467.55) µg·mL-1 96 h after the last treatment. These levels were higher than the minimum inhibitory concentrations-namely, between 4 and 64 µg·mL-1-against pathogenic isolates Trueperella pyogenes, Streptococcus intermedius, Escherichia coli and Bacillus pumilis. Higher concentrations over time were measured in milk rather than in plasma, indicating the need to monitor the withdrawal time for milk. The intrauterine infusion of oxytetracycline for three consecutive days resulted in the successful treatment of metritis in terms alleviating inflammation and restoring the estrus cycle in cows.

Comparison between Some Phenotypic and Genotypic Methods for Assessment of Antimicrobial Resistance Trend of Bovine Mastitis Staphylococcus aureus Isolates from Bulgaria.

The aim of this study was to assess the resistance of bovine mastitis S. aureus isolates from farms in Bulgaria to different classes of chemotherapeutic drugs by comparison of some phenotypic and genotypic methods by means of Cohen's kappa statistics. The study comprised 546 milk samples from subclinical and clinical mastitis at 14 farms from 9 districts in the country. A total of 92 Staphylococcus aureus strains were isolated from tested samples and identified by nuc PCR. The results demonstrated high levels of resistance to sulfadimethoxine (87%), followed by resistance to penicillin (33.7%), erythromycin (13%), streptomycin (8.7%), tetracycline (6.5%) and gentamicin (1.1%). The comparison of both phenotypic tests with respect to 9 antimicrobials revealed strong agreement with kappa coefficient 0.836. An almost complete agreement was evidenced between phenotypic resistance to penicillin and blaZ gene presence, to methicillin with mecA gene, to tetracycline with tet genes, but the agreement between erythromycin resistance and erm genes presence was moderate. This study was the first to demonstrate discrepancy between the behaviour to cefoxitin in the disk diffusion test and oxacillin in the MIC test for an isolate shown to carry the mecA gene in the subsequent genetic analysis. Considering the detected discrepancies for some of isolates, an integral evaluation through phenotypic and molecular methods for monitoring of antimicrobial resistance of Staphylococcus aureus is recommended.

The use of ultrasonographic measurement of the heart size and fetal heart rate variation for gestational age determination in local Bulgarian goats.

BACKGROUND: Determination of gestational age in goats was performed using ultrasound measurements of different fetal biometric parameters. There are no data showing quadratic and exponential dependence between fetal heart parameters and gestational age. OBJECTIVES: The objective of the present study was to test the significance of the defined indicators (fetal heart rate, longitudinal and transverse fetal heart axes) for determining gestational age in local Bulgarian goats. METHODS: A total of 24 pregnant local Bulgarian goats, aged between 2 and 5 years, body weight 42-50 kg were included in the study. Ultrasonographic examinations were performed weekly on gestational days 21, 28, 35, 42 and 49. After gestational day 49 until parturition, scans were performed biweekly. The data from the measurements were subjected to linear (y = a + bx), quadratic (y = aх2 + bх + c) and exponential (y = axn ) correlation. RESULTS: Average fetal heart rate values decreased with pregnancy progression. The coefficient of determination (R2 ) and standard error of estimate (SEE) of the heart rate indicator were 0.72, 0.75, 0.58 and 15.1, 14.4, 19.2 days, respectively, for the three correlations. Longitudinal diameter (long axis) had the following values for R2 = 0.94, 0.96, 0.96 and for SEE = 7.0, 5.5, 5.5 days, for the linear, quadratic and exponential correlations, respectively, while the values of the fetal heart transverse diameter (short axis) were higher than those of the external one (R2 = 0.95, 0.97, 0.97). Simultaneously, SEE is lower (SEE = 6.1, 4.9, 5.0 days) compared to that found for the long heart fetal axis. CONCLUSIONS: For precise estimation of gestational age, use of longitudinal (long) and transverse (short) axes is recommended. Fetal heart rate is not an exact indicator but can be used as a reference for gestational age along with changes in the heart size and echogenicity.

Doppler ultrasound measurements of the blood flow velocity in the fetal heart and aorta in Bulgarian White milk goats.

The aim of the present study was to determine the parameters characterising the blood flow velocity in the fetal heart and aorta in goats by Doppler ultrasonography. Twenty-four Bulgarian White milk goats, aged 4-6 years, weighing 45-51 kg were used in the study. Doppler ultrasound measurements of the blood flow velocity in fetal heart were done at the end of first, second and third trimesters of pregnancy, and in the fetal aorta - at the end of the second and third trimesters of pregnancy. Ultrasound investigations were carried out by transabdominal approach with linear and convex transducers, with frequency 8.0 MHz. Blood flow parameters that included maximum and minimum systolic velocity, end-diastolic velocity, pulsatility index, resistance index and systolic/diastolic ratio were measured by spectral Doppler ultrasonography. The results showed that the highest maximum systolic velocity in the fetal heart was observed at the end of the first pregnancy trimester. Statistically significantly (p < 0.05) lower values of this parameter were registered in the second and third pregnancy trimesters. A similar tendency was observed for minimum systolic velocity, end-diastolic velocity and systolic/diastolic ratio. The pulsatility index and resistance index had similar values for the three studied periods. Examination of the fetal aorta showed statistically significant differences (p < 0.05) of minimum systolic velocity, end-diastolic velocity, pulsatility and resistance index in the third, compared with the second trimester of pregnancy. The analysis of the results suggested that Doppler ultrasonography can be used to monitor fetal blood flow changes during the various stages of pregnancy.

Resumption of Cyclic Ovarian Activity by Herbal Preparation AyuFertin in Bulgarian Murrah Buffaloes at Early Postpartum.

This study evaluates the phytochemical composition and ability of herbal preparation AyuFertin, supplemented shortly after calving, to resume the cyclic ovarian activity in postpartum Bulgarian Murrah buffaloes. In total, 13 healthy Bulgarian Murrah buffaloes at the age of 4-10 years, calved in March-April 2019 were involved in the experiment. Seven experimental buffaloes were treated individually per os with AyuFertin according to producer instruction. All animals were subjected to regular ultrasound examination of ovaries. GC-MS analysis of fatty acids composition and HPLC-DAD analysis of carotenoid and tocopherol content in AyuFertin were conducted. The determination of estrus occurrence and natural mating were done by a fertile bull. The development of the large follicles (12.5-14.3 mm) in 85.7% of treated animals was observed on day 24 postpartum compared to 0% in controls. Clinical signs of estrus were recorded in 71.4% of the animals, followed by an 80% pregnancy rate versus 16% in controls within 70 days after calving. In conclusion, the supplementation of the bioactive herbal product AyuFertin from day 21 postpartum can stimulate the follicular growth in the buffalo's ovary, which leads to the development of follicles with ovulatory capacity, followed by successful ovulation. The above-mentioned treatment resulted in a better pregnancy rate in the treated multiparous compared to the control buffaloes bred by natural service.

Oxytetracycline Pharmacokinetics After Intramuscular Administration in Cows with Clinical Metritis Associated with Trueperella Pyogenes Infection.

Systemic therapy with oxytetracycline is often used for treatment of clinical metritis although data about its penetration into the uterus and uterine secretion are lacking. Uterine secretions and milk from six cows with clinical metritis were collected for microbiological assay. The animals were treated intramuscularly with long-acting oxytetracycline (20 mg/kg) and samples of plasma, milk and uterine secretions were collected for determination of the antibiotic concentrations by HPLC-PDA analysis. Pharmacokinetics of the antibiotic and in silico prediction of its penetration into the uterus were described. Trueperella pyogenes with MIC values of 16-64 µg mL-1 was isolated (n of cows = 4) from uterine secretions. Oxytetracycline showed fast absorption and penetration in the uterine secretions and milk. No change of withdrawal time for milk was necessitated in cows with clinical metritis. Maximum levels in uterine secretions and predicted concentrations of oxytetracycline in the uterus were lower than MIC values. Systemic administration of long-acting oxytetracycline did not guarantee clinical cure and was not a suitable choice for treatment of clinical metritis associated with Trueperella pyogenes. The appropriate approach to antibiotic treatment of uterine infections of cows requires knowledge on penetration of the antibiotics at the site of infection and sensitivity of pathogens.

Determination of the gestational age through ultrasound measurements of some uterine and foetal parameters in Bulgarian local goats.

The aim of the current study was to determine the gestational age in goats through the measurement of some foetal and uterine structures. Twenty-four pregnant Bulgarian local goats were submitted to ultrasound foetometry by transrectal and transabdominal approaches. Ultrasound measurement of studied parameters was done at 7-day intervals during the first trimester of gestation (days 21-49) and at 14-day intervals between gestation days 49 and 133. Associations of foetometry data with foetal age were investigated by linear (y = a + bx), quadratic (y = ax2  + bx + c) and exponential (y = axn ) equations. According to the results, some biometric parameters (uterine lumen diameter, crown-rump length, trunk diameter, biparietal diameter, foetal aortic diameter) exhibited a stronger correlation to gestational age (R2  ≥ 0.90) and lower standard error of the estimate (SEE ≤ 8) as compared to foetal eye orbit diameter, and outer and inner placentome diameters (R2  ≤ 0.90; SEE ≥ 8). Each of these biometric parameters could be measured in different periods of gestation, thus allowing for monitoring and determination of foetal age at almost all stages of pregnancy.