Biochemical Composition Of Follicular Fluid Research Articles

Evaluation of cortisol, thyroxine (T3) and triiodothyronine (T4) concentrations in preovulatory follicles in jennies.

Reference values of follicular fluid components at different stages of follicular development in jennies are unknown. It is important to estimate such values because the biochemical composition of follicular fluid provides a suitable microenvironment for oocyte development, growth and maturation. The experimental protocol was carried out in the Equine Reproduction Clinic of the FMVZ-UNAM and was approved by an Institutional Sub-committee for the Care and Use of Experimental Animals (SICUAE.MMVZ-2020/4-2). Seven healthy female donkeys “Mexican burro” with average body condition score of 3 on a scale of 1 to 5 (Svendsen et al. The professional Handbook of the Donkey. 2008), were monitored by transrectal ultrasonography through the Veterinary Digital Diagnostic Ultrasound System 5 MHz (Model, DUS 60 VET) with a multi-frequency linear transducer. The ovum pick up (OPU) by transvaginal aspiration (TVA) was performed in a total of 70 preovulatory follicles, 10 for each jenny using a real- time B- mode ultrasound vaginal 6.5 MHz (Sonovet 600) with curvilinear transducer and 12 gauge (12 G) needle. Follicular fluid components were measured and analyzed by Luminex Multiplex Assays at the Universidad Nacional Autónoma de México. The relationship between cortisol, thyroxine (T3) and triiodothyronine (T4) concentrations in preovulatory follicles and follicular size was assessed. The preovulatory follicles are classified in three groups accordingto the diameter: small (27-30 mm), medium (31-35 mm) and large (36-38 mm). Concentrations of cortisol were lower in the large follicles group (18.8 ng/ml) compared with small (721.44 ng/ml) and medium follicles (361 ng/ml). The concentrations of thyroxine were higher in the medium follicle group (49 ng/ml) than in the small (21ng/ml) and large follicle groups (15.9 ng/ml). Finally, the concentration of triiodothyronine was higher in the small follicles group (1043 ng/ml) followed by the medium (545 ng/ml) and the lowest concentration was in the large follicles group (22 ng/ml). It is concluded that thyroxine concentrations are lower than T4 concentrations in preovulatory follicles irrespective of size as reported for human follicular fluid (Wakim. et. al. Fertil Steril 1993; 59(6) 1187–1190). Furthermore, no relationship was found between preovulatory follicular size and thyroxine hormone concentrations. It has been suggested that cortisol and triiodothyronine concentrations are related to preovulatory follicle size. Cortisol concentrations are higher in preovulatory follicles that in subordinate follicles and cortisol is a mediator of apoptosis in granulosa cells, differentiation, development and maturation of tissues, specifically in the oocyte (Acosta & Miyamoto. Anim Reprod Sci 2004; 82:127-140, Scarlett. et al. Dom Anim Endocrinol 2016; 59:11-22). The role of cortisol within the different sizes of preovulatory follicles is unknown.

Comparative Analysis of Biochemical Constituents between Ovarian Follicular Fluid and Serum in Ganjam Goat (Capra hircus)

Background: Follicular fluid is in part exudates of serum and is also partially composed of locally produced substances, which are related to the metabolic activity of the follicular cells. The knowledge of the biochemical composition of follicular fluid can provide useful information about the requirements, growth and maturation of oocyte and which may be used as a provisional guide for formulating suitable culture media for in vitro cell culture and maturation in a particular species. Methods: Follicular fluid were aspirated from small follicles ( less than 3 mm; SF), medium follicles (3-5 mm; MF) and large follicles ( greater than 5 mm; LF). Blood samples were also collected for extraction of serum. Follicular fluid and serum samples were analyzed for biochemical constituents. Conclusion: It can be concluded that some biochemical constituents of follicular fluid such as glucose, cholesterol, total protein and globulin were higher in large follicles compared to small follicles. Moreover, serum levels of most of biochemical constituents were higher than their levels in different sizes of follicular fluid. This present investigation of comparative analysis of biochemical constituents between serum and ovarian follicles in Ganjam goat of Odisha will provide clues to researchers who work on reproductive physiology in different breeds of goat.

96 Follicular Fluid and Serum Biochemical Composition in Alpacas (Vicugna pacos) with Different Nutritional Planes

The importance of nutrition on composition of follicular fluid and oocyte quality is recognised in several species but limited information is available on alpacas and would explain the high rate of early embryo mortality. The aim of this study was to analyse the biochemical composition of serum and follicular fluid (FF) from alpacas (Vicugna pacos) with different nutritional planes. Twelve adult female alpacas between 6 and 8 years old were assigned randomly to either a high (n = 6) or low (n = 6) plane of nutrition. Nutritional planes were defined by the grazing condition of the natural pasture with (high) or without (low) supplementation with 200 g of concentrate/day (total digestible nutrients: 52%, protein: 16%, and metabolizable energy: 2.8 Mcal kg−1). The nutritional conditions were imposed 1 month before the start of the experiment.The concentrations of total glucose, cholesterol, triglycerides, and proteins were determined with a semi-automatic biochemical analyzer (SINOWA, China). When an ovulatory follicle ≥7 mm was detected by ultrasonography using a 5-MHz linear-array transducer (SSD-500, Aloka, Tokyo, Japan), a transvaginal transducer with an attached needle guide (UST-945BP-5, Aloka) was used to collect follicular fluid. Follicular puncture was performed using a disposable 19-gauge # 12 mm hypodermic needle connected to a 50-mL conical tube via a silicon tube. A caudal epidural anaesthesia was induced with 4 mL of 2% lidocaine. Blood samples were obtained by jugular venipuncture into serum vacutainers and kept at room temperature for 30 min. Follicular fluid and serum were centrifuged at 1500 × g for 20 min, decanted, and stored at –20°C until analysed. Results were analysed by Student’s t-test. The results are presented in Table 1. The results suggest differences in biochemical composition of glucose and cholesterol in follicular fluid, which could explain the oocyte quality and embryo survival in alpacas. Table 1.Biochemical composition of follicular fluid (FF) and serum from alpacas (Vicugna pacos) under 2 nutritional planes Research was supported by INNOVATE PERU Grant 405-PNICP-PIAP-2014.

111 BIOCHEMICAL COMPOSITION OF FOLLICULAR FLUID IN RELATION TO THE STIMULUS TO INDUCE OVULATION IN ALPACAS (VICUGNA PACOS)

Alpacas, like other camelids, are induced ovulators. A study was designed to determine the effect of the ovulation-inducing stimulus on the biochemical composition of follicular fluid. Adult female alpacas (n = 18) were examined daily for 3 days by transrectal ultrasonography using a 5-MHz linear-array transducer (Aloka SSD-500, Tokyo, Japan). When the largest growing ovarian follicle was ≥7 mm, alpacas were given 1.0 mL of seminal plasma intramuscularly (SP, n = 9) or 40 µg of busereline acetate intramuscularly (GnRH, n = 9). A transvaginal transducer with an attached needle guide (Aloka UST-945BP-5) was used for collection of follicular fluid 22 h post-induction. Follicular contents were then centrifuged at 800 × g for 20 min to separate the fluid from the cells. The follicular fluid was collected and stored at –20°C until analysis with a semi-automatic biochemical analyzer (SINOWA, China). The results were glucose 49.17 and 47.95 (mg/dL; P > 0.05), total protein 1.85 and 1.15 (g/dL; P < 0.05), albumin 1.11 and 1.13 (g/dL; P > 0.05), triglycerides 3.94 and 3.16 (mg/dL; P > 0.05), cholesterol 39.01 and 42.5 (mg/dL; P > 0.05), phosphatase 32.68 and 21.36 (IU/L; P < 0.05), alanine aminotransferase 3.66 and 5.07 (IU/L; P > 0.05), and lactate dehydrogenase 42.17 and 27.27 (IU/L; P > 0.05) for SP or GnRH treatments, respectively. Results suggest the need to continue research to explain the effect of possible differences in total protein, cholesterol, and phosphatase on oocyte-expressed genes and follicular development. Research was supported by the project no. 405-PNICP-PIAP-2014-UNMSM.

Follicular Fluid Concentrations of Metabolic Stressors in Normal, Obese, Metabolic Stressed and Emaciated Ewes

Somatic cell quality and oocyte quality may be influence by follicular fluid composition. The changes in metabolic composition in follicular fluid reflect the changes in biochemical composition of follicular fluid. The aim of this study was to examine the metabolic stressors composition of follicular fluid of sheep (obese, normal, metabolic stressed and emaciated). Follicular fluid samples were assayed for cholesterol, triglycerides, urea, ammonia, non-esterified fatty acids and β-hydroxybutryic acids. Metabolic stressors like total NEFA, urea and beta-hydroxy butyrate concentrations were significantly higher and cholesterol and triglycerides concentrations were significantly lowered in follicular fluid of metabolic stressed and emaciated ewes.

Changes in biochemical composition of follicular fluid during reproductive acyclicity in water buffalo ( Bubalus bubalis)

This study describes the changes in biochemical composition of follicular fluid during reproductive acyclicity in buffalo. A total of 73 pairs of ovaries collected from 26 reproductively acyclic and 47 reproductively cyclic buffaloes were used in the investigation. Ovarian follicles were classified into small (5.0–6.9 mm), medium (7.0–9.9 mm) and large (≥10.0 mm) sized categories depending upon their diameter. Follicular fluid was aspirated, processed and assayed for glucose, cholesterol, total protein, acid phosphatase and alkaline phosphatase. Glucose concentration was lesser in reproductively acyclic compared to cyclic buffaloes (19.3 ± 2.59 mg/dl compared to 32.6 ± 2.60 mg/dl; P < 0.05), mainly due to difference in concentration between small sized follicles (12.4 ± 2.59 mg/dl compared to 28.0 ± 3.32 mg/dl; P < 0.05). Cholesterol concentration was also lesser in reproductively acyclic compared to cyclic buffaloes (32.2 ± 2.14 mg/dl compared to 35.5 ± 2.16 mg/dl; P < 0.05) and this was related to the lesser concentration found in large follicles (13.8 ± 3.45 mg/dl compared to 37.2 ± 4.10 mg/dl; P < 0.001). Total protein and acid phosphatase levels were not affected by either the reproductive cyclicity status or the follicular size (4.9 ± 1.07 g/dl to 6.0 ± 0.28 g/dl and 1.2 ± 0.17 U/dl to 2.5 ± 1.22 U/dl, respectively). An increased alkaline phosphatase activity was, however, observed in reproductively acyclic compared to cyclic buffaloes (27.5 ± 3.08 U/dl compared to 14.0 ± 1.09 U/dl; P < 0.0001). In conclusion, results of the present study indicate an alteration in the biochemical composition of follicular fluid during reproductive acyclicity in buffalo. The findings provide further support to the notion that poor nutrition is an important factor triggering reproductive acyclicity in buffalo.

Biochemical composition of blood plasma and follicular fluid in relation to follicular size in buffalo

The aim of this study was to examine the biochemical composition of follicular fluid from different-sized follicles and its relationship with that of blood plasma in buffalo. Ovaries of adult and healthy buffaloes were collected after slaughter. Follicular fluid was aspirated from three size classes of follicles (4–5, 6–9, and 10–20 mm diameter). Blood samples were also collected from these buffaloes immediately before slaughter. The follicular fluid and blood plasma samples were analyzed for metabolites (glucose, total protein, albumin, globulin, cholesterol, triglycerides, urea, and creatinine), ions (calcium and phosphorus), and enzymes (alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase). The follicular fluid calcium, urea, creatinine, albumin, and ASAT and ALAT concentrations were not affected by the size of the ovarian follicles. Follicular fluid concentrations of phosphorus, cholesterol, triglyceride, ALK, and LDH decreased from small to large follicles. Phosphorus, albumin, and LDH concentrations in plasma were significantly lower than the levels in all follicle classes. The plasma concentrations of glucose, creatinin, cholesterol, triglyceride, protein, and glubolin were higher than in the small, medium, and large follicles. The concentration of glucose in the small follicles was significantly lower than in the medium and large follicles. Total protein concentration in fluid of small follicles was significantly higher than in the large follicles. The amount of globulin in medium follicles was higher than in the small and large follicles. The plasma concentration of ALK was significantly lower than in the small and medium follicles. ALK concentration difference between large follicles and plasma was not significant.

Metabolite and ionic composition of follicular fluid from different-sized follicles and their relationship to serum concentrations in dairy cows

Metabolic changes in blood serum may be reflected in the biochemical composition of follicular fluid and could indirectly influence oocyte quality. The purpose of this study was to examine the biochemical composition of follicular fluid harvested from different-sized follicles and its relationship with that of blood serum in dairy cattle. Following slaughter, blood samples were collected from dairy cows ( n=30) and follicular fluid aspirated from three size classes of non-atretic follicles (<4 mm, 6–8 mm and >10 mm diameter). Samples remained independent between cows and between size classes within cows. Serum and follicular fluid samples were assayed using commercial clinical and photometric chemistry assays for ions (sodium, potassium and chloride) and metabolites (glucose, β-hydroxybutyrate (β-OHB), lactate, urea, total protein, triglycerides, non-esterified fatty acids (NEFA) and total cholesterol). Results showed that follicular fluid concentrations of glucose, β-OHB and total cholesterol increased from small to large follicles and decreased for potassium, chloride, lactate, urea and triglycerides. There was a significant concentration gradient for all variables between their levels in serum and follicular fluid ( P<0.05). Significant correlations were observed for chloride ( r=0.40), glucose ( r=0.56), β-OHB ( r=0.85), urea ( r=0.95) and total protein ( r=0.60) for all three follicle size classes and for triglycerides ( r=0.43), NEFA ( r=0.50) and total cholesterol ( r=0.42) for large follicles ( P<0.05). The results from the present study suggest that the oocyte and the granulosa cells of dairy cows grow and mature in a biochemical environment that changes from small to large follicles. Furthermore, the significant correlation between the composition of serum and follicular fluid for the above-mentioned metabolites suggests that metabolic changes in serum levels will be reflected in the follicular fluid and, therefore, may affect the quality of both the oocyte and the granulosa cells.