Miniaturized PVC Sensors for Determination of Fe2+, Mn2+ and Zn2+ in Buffalo-Cows’ Cervical Mucus Samples

Three polyvinyl chloride membrane sensors were developed for the electrochemical evaluation of ferrous, manganese and zinc ions. The sensors were used for assaying metal ions in cervical mucus (CM) of Egyptian river buffalo-cows (Bubalus bubalis) as their levels vary dependent on cyclical hormone variation during different phases of estrus cycle. The presented sensors are based on using ionophores, β-cyclodextrin (β-CD), hydroxypropyl β-cyclodextrin (HP-β-CD) and sulfocalix-4-arene (SCAL) for sensors 1, 2 and 3 for Fe2+, Mn2+ and Zn2+, respectively. Dioctyl phthalate (DOP) was used as the plasticizer in a polymeric matrix of polyvinylchloride (PVC). For increasing the selectivity and sensitivity of the sensors, each sensor was enriched with a suitable complexing agent, which enhanced the sensor’s response. For sensor 1, β-CD was mixed with bathophenanthroline; for sensor 2, porphyrin was incorporated with HP-β-CD; while for sensor 3, oxine was the used complexing agent with SCAL. Linear responses of 10-7-10-2 M with cationic slopes of 53.46, 45.01 and 50.96 over pH range 4-8 were obtained using coated graphite sensors for ferrous, manganese and zinc ionic solutions, respectively. The three sensors were validated, according to the IUPAC guidelines. The obtained results by the presented potentiometric procedures were statistically analyzed and compared with those obtained by atomic absorption spectrophotometric method (AAS). No significant differences for either accuracy or precision were observed between the two techniques. Successful application for the determination of the three studied cations in CM, for the purpose to determine the proper time for artificial insemination (AI) was achieved. The results were compared with those obtained upon analyzing the samples by AAS. Proper detection of estrus and correct time of AI was necessary to maximize the production of buffaloes. In this experiment, 30 multi-parous buffalo-cows were in second to third lactation and weighting 415-530 kg, and were synchronized with OVSynch protocol. Samples were taken in three times around ovulation, on day 8 of OVSynch protocol, on day 9 (20 h before AI) and on day 10 (1 h before AI). Beside analysis of trace elements (Fe2+, Mn2+ and Zn2+) in CM using the three sensors, the samples were analyzed for the three cations and also Cu2+ by AAS in the CM samples and blood samples. The results obtained were correlated with hormonal analysis of serum samples and ultrasonography for the purpose of determining of the optimum time of AI. The results showed significant differences and powerful correlation with Zn2+ composition of CM during heat phase and the ovulation time, indicating that the parameter could be used as a tool to decide optimal time of AI in buffalo-cows.

Enzymes Activity in Bovine Cervical Mucus Related to the Time of Ovulation And Insemination

Forty-five dairy cows were used to compare the enzyme activity of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), α -amylase in the cervical mucus of cows during spontaneous and induced estrus using progestagen or PGF2 α and to determine whether these enzymes affect the fertility in cows with induced estrus, at the time of Al. The animals were assigned to 3 groups (no treatment, a Crestar® for 12 days, a double im injection of PGF2 α). The cows were artificially inseminated (AI). Cervical mucus samples were collected from all cows 3 to 5 min before the AI. The results are summarized as follows: ALP and α -amylase activity for spontaneous estrus were similar to those for induced estrus (P>0.05) . LDH activity levels during spontaneous and PGF2 α induced estrus was significantly lower (P < 0.001) than that in progestagene induced estrus groups. While no difference was found between the first and the third groups. Our result showed a significant difference in LDH activity levels between cows conceived with 2 or more AI and those conceived with 1 AI. The result of this study showed that the enzyme activity in cervical mucus is helpful for detection of ovulation and time of AI.