The aim of this study was to investigate the period of estrus cycle in aceh cattle, Indonesia, based on vaginal cytology techniques. Four healthy females of aceh cattle with average weight of 250–300 kg, age of 5–7 years, and body condition score of 3-4 were used. All cattle were subjected to ultrasonography analysis for the occurrence of corpus luteum before being synchronized using intramuscular injections of PGF2 alpha 25 mg. A vaginal swab was collected from aceh cattle, stained with Giemsa 10%, and observed microscopically. Period of estrus cycle was predicted from day 1 to day 24 after estrus synchronization was confirmed using ultrasonography analysis at the same day. The result showed that parabasal, intermediary, and superficial epithelium were found in the vaginal swabs collected from proestrus, metestrus, and diestrus aceh cattle. Proportions of these cells in the particular period of estrus cycle were 36.22, 32.62, and 31.16 (proestrus); 21.33, 32.58, and 46.09 (estrus); 40.75, 37.58, and 21.67 (metestrus); and 41.07, 37.38, and 21.67 (diestrus), respectively. In conclusion, dominant proportion of superficial cell that occurred in estrus period might be used as the base for determining optimal time for insemination.
This study was conducted to validate a commercial testosterone enzyme-linked immunosorbent assay (ELISA) kits (DRG EIA-1559) inanalytic and biological manner for measuring serum testosterone concentrations in kacang goats. This study used 18 healthy kacang goats, sixbucks (>2 years), six kids (<6 months), and six does (>2 years). Blood samples were collected from jugular vein and prepared as serum. Twovalidation tests were performed, an analytical validation comprises a parallelism, accuracy, precision and sensitivity and a biological validationby comparing testosterone concentration from bucks, kids, and does. Testosterone concentrations were measured using ELISA technique. Data ofanalytical validation were analyzed descriptively and test of equality of slope was performed to see the parallelism between samples and standardcurves. Analysis of variance (ANOVA) was used for biological validation data. Results of parallelism showed that sample curve was parallel tothe standard curve. Accuracy, precision (% CV of intra-and inter-assay) and sensitivity of the assay were 99.65±4.27%, <10%, <15% and 0.083ng/ml, respectively. Results of biological validation showed that the assay used were accurately measured testosterone which testosteroneconcentrations in bucks were significantly higher compared to kids and does (P<0.05). In conclusion, a commercial testosterone ELISA kits(DRG EIA-1559) is a reliable assay for measuring serum testosterone concentration in kacang goats. Key words: analytical and biological validations, ELISA, testosterone, kacang goat
Background: To obtain accurate measurements of cortisol (C) and testosterone (T) in Aceh cattle, commercial enzyme-linked immunosorbent assay (ELISA) kits need to be carefully validated. Moreover, repeated freeze-thaw cycles during the storage of the samples may affect the stability of the hormones in the serum. Here, we test the reliability of C and T concentration measurements in the serum of Aceh cattle, obtained using commercial C and T ELISA kits designed to measure human C and T concentrations. Further, we evaluate the effect of repeated freeze-thaw cycles on the stability of C and T concentrations in the serum. Methods: Commercial C (Cat. no. EIA-1887) and T (Cat. no. EIA-1559) ELISA kits from DRG Instruments GmbH were validated through an analytical validation test (i.e., parallelism, accuracy, and precision) and a biological validation test (for C: effect of transportation on the C excretion; for T: the concentrations of T between bulls and cows). To test the effects of freeze-thaw cycles, cattle serum was subjected to the following treatments: (i) remained frozen at -20OC (control group); (ii) exposed to freeze-thaw cycles for two, four, six, and eight times (test groups). Results: Parallelism, accuracy, and precision tests showed that both C and T ELISA kits adequately measured C and T in the serum of Aceh cattle. Concentrations of C post-transportation were significantly higher than pre-transportation (p<0.05). Concentrations of T in bulls were significantly higher than in cows (p<0.05). After four to eight freeze-thaw cycles, C concentrations were significantly lower compared to the control group (all p < 0.05). In contrast, T concentrations remained stable (all p>0.05). Conclusions: Commercial C (EIA-1887) and T (EIA-1559) ELISA kits are reliable assays for measuring serum C and T, respectively, in Aceh cattle. Repeated freeze-thaw cycles significantly affected the stability of serum C, but did not for T.
This study was conducted to examine testosterone concentrations its relationship with the scrotal circumference and physical characteristics of semen in aceh bulls. Semen samples were collected weekly from jugular vein of three aceh bulls aged 4-5 years old for 10 weeks. Testosterone concentration was measured by enzyme-linked immunosorbent assay (ELISA) method. Semens were collected by using artificial vagina and evaluated for physical characteristics namely ejaculatory volume, pH, and sperm motility, concentration, and abnormalities. Data were analyzed using correlation-regresion test. Testosterone concentrations showed a positive correlation with scrotal circumference (r = 0.799), number of sperm (r = 0.703), sperm motility (r = 0.857) and sperm abnormalities (r = -0,877). No correlation, however, was found between testosterone concentrations with semen volume (r = 0.038) and pH (r = 0.418). It can be concluded that testosterone concentrations correlated positively with scrotal circumference, numbers of sperm, sperm of motility and sperm of abnormality.
Background: To obtain accurate measurements of cortisol (C) and testosterone (T) in Aceh cattle, commercial enzyme-linked immunosorbent assay (ELISA) kits need to be carefully validated. Moreover, repeated freeze-thaw cycles during the storage of the samples may affect the stability of the hormones in the serum. Here, the reliability of C and T concentration measurements in the serum of Aceh cattle, was tested using commercial C and T ELISA kits designed to measure human C and T concentrations. Further, the effect of repeated freeze-thaw cycles on the stability of C and T concentrations in the serum was evaluated. Methods: Commercial C (Cat. no. EIA-1887) and T (Cat. no. EIA-1559) ELISA kits from DRG Instruments GmbH were validated through an analytical validation test (i.e., parallelism, accuracy, and precision) and a biological validation test (for C: effect of transportation on the C secretion; for T: the concentrations of T between bulls and cows). To test the effects of freeze-thaw cycles, cattle serum was subjected to the following treatments: (i) remained frozen at -20OC (control group); (ii) exposed to freeze-thaw cycles for two, four, six, and eight times (test groups). Results: Parallelism, accuracy, and precision tests showed that both C and T ELISA kits adequately measured C and T in the serum of Aceh cattle. Concentrations of C post-transportation were significantly higher than pre-transportation (p<0.01). Concentrations of T in bulls were significantly higher than in cows (p<0.01). After four to eight freeze-thaw cycles, C concentrations were significantly lower compared to the control group (all p < 0.05). In contrast, T concentrations remained stable (all p>0.05). Conclusions: Commercial C (EIA-1887) and T (EIA-1559) ELISA kits are reliable assays for measuring serum C and T, respectively, in Aceh cattle. Repeated freeze-thaw cycles significantly affected the stability of serum C, but did not for T.
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