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

2018 ◽  
Vol 30 (1) ◽  
pp. 187
Author(s):  
W. Huanca ◽  
F. Hilari ◽  
M. Ticona ◽  
B. Lira ◽  
J. C. Villanueva ◽  
...  

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.

2017 ◽  
Vol 29 (1) ◽  
pp. 164 ◽  
Author(s):  
W. Huanca ◽  
A. Castro ◽  
N. Gomez ◽  
A. Cordero

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.


Reproduction ◽  
2020 ◽  
Vol 159 (5) ◽  
pp. 585-599
Author(s):  
Zaramasina L Clark ◽  
Derek A Heath ◽  
Anne R O’Connell ◽  
Jennifer L Juengel ◽  
Kenneth P McNatty ◽  
...  

Ewes with single copy mutations in GDF9, BMP15 or BMPR1B have smaller preovulatory follicles containing fewer granulosa cells (GC), while developmental competency of the oocyte appears to be maintained. We hypothesised that similarities and/or differences in follicular maturation events between WT (++) ewes and mutant ewes with single copy mutations in BMP15 and BMPR1B (I+B+) are key to the attainment of oocyte developmental competency and for increasing ovulation rate (OR) without compromising oocyte quality. Developmental competency of oocytes from I+B+ animals was confirmed following embryo transfer to recipient ewes. The microenvironment of both growing and presumptive preovulatory (PPOV) follicles from ++ and I+B+ ewes was investigated. When grouped according to gonadotropin-responsiveness, PPOV follicles from I+B+ ewes had smaller mean diameters with fewer GC than equivalent follicles in ++ ewes (OR = 4.4 ± 0.7 and 1.7 ± 0.2, respectively; P < 0.001). Functional differences between these genotypes included differential gonadotropin-responsiveness of GC, follicular fluid composition and expression levels of cumulus cell-derived VCAN, PGR, EREG and BMPR2 genes. A unique microenvironment was characterised in I+B+ follicles as they underwent maturation. Our evidence suggests that GC were less metabolically active, resulting in increased follicular fluid concentrations of amino acids and metabolic substrates, potentially protecting the oocyte from ROS. Normal expression levels of key genes linked to oocyte quality and embryo survival in I+B+ follicles support the successful lambing percentage of transferred I+B+ oocytes. In conclusion, these I+B+ oocytes develop normally, despite radical changes in follicular size and GC number induced by these combined heterozygous mutations.


Author(s):  
Özgür BİGE ◽  
Bülent GÜLEKLİ ◽  
Ahmet DEMİR ◽  
Funda GÖDE ◽  
Semra KOÇTÜRK ◽  
...  

2021 ◽  
Vol 64 (1) ◽  
Author(s):  
Jianyu Li ◽  
Xin Zhao ◽  
Laura S. Bailey ◽  
Manasi N. Kamat ◽  
Kari B. Basso

AbstractThe biochemical composition of organic fertilizers largely determines their nutrient supply characteristics following soil application as well as their potential impact on soil microbial communities. Yet, limited information is available regarding the biochemical composition of organic fertilizers derived from different nutrient sources. Here, we qualitatively analyzed the presence and abundance of proteins, lipids, and metabolites in a liquid fish fertilizer (LFF) product and a type of granular organic fertilizer (GOF) commonly used in organic vegetable production, using liquid chromatography–tandem mass spectrometry (LC–MS/MS). Our results suggest that the presence and abundance of proteins, lipids, and metabolites differ greatly between GOF and LFF. The qualitative analysis shows LFF as a rich source of metabolites, while complex proteins and long-chain saturated fatty acids are dominant in GOF. The degree of biochemical composition complexity may help explain the varying impacts of different types of organic fertilizers on nutrient availability, soil health, and environmental quality.


2018 ◽  
Vol 22 (4) ◽  
pp. 188
Author(s):  
Simon P. Ginting ◽  
Kiston Simanihuruk ◽  
Antonius Antonius ◽  
Andi Tarigan

The aim of this study was to evaluate the growth of and feed utilization by Boer x Kacang crosses goats fed on total mixed ration differing in protein and energy levels. Four total mixed rations combination were formulated to contain 16 and 18% crude protein and 2650 and 2850 Kcal ME/ kg DM (dried matter) energy density. Twenty-eight male Boer x Kacang crosses goats (14.5 ± 1.14 kg) and of age ranging from 4 to 5 months were randomly allocated to one of these four TMRs (total mixed rations) (7 animals/TMR). The effects of dietary treatment were assessed using the general linear model and significance of the diet effects was detected using Duncan’s multiple range test. Dry matter intake increased as metabolizable energy density of diet increased from 2650 to 2850 Kcal/kg DM, but it is not affected by increasing crude protein level from 16 to 18%. The average daily gains were not improved (P&gt;0.05) as the crude protein levels and metabolizable energy density of diet increased. Crude protein levels and ME density did not affect (P&gt;0.05) the DM, OM and energy digestibility, but NDF digestibility was affected by the ME density of diets (P&lt;0.05). Daily N intakes were greater (P&lt;0.0%) in goats received diets higher in the crude protein and metabolizable energy levels. At this high rate of feed intake this type of goats are able to gain optimally when offered feed with crude protein level of 16% and metabolizable energy density of 2850 Kcal/kg DM.


2020 ◽  
Vol 7 ◽  
Author(s):  
Jennifer C. Lutz ◽  
Susan L. Johnson ◽  
Kimberly J. Duprey ◽  
Paul J. Taylor ◽  
Henry William Vivanco-Mackie ◽  
...  

The alpaca (Vicugna pacos) is an important species for the production of fiber and food. Genetic improvement programs for alpacas have been hindered, however, by the lack of field-practical techniques for artificial insemination and embryo transfer. In particular, successful techniques for the cryopreservation of alpaca preimplantation embryos have not been reported previously. The objective of this study was to develop a field-practical and efficacious technique for cryopreservation of alpaca preimplantation embryos using a modification of a vitrification protocol originally devised for horses and adapted for dromedary camels. Four naturally cycling non-superovulated Huacaya females serving as embryo donors were mated to males of proven fertility. Donors received 30 μg of gonadorelin at the time of breeding, and embryos were non-surgically recovered 7 days after mating. Recovered embryos (n = 4) were placed individually through a series of three vitrification solutions at 20°C (VS1: 1.4 M glycerol; VS2: 1.4 M glycerol + 3.6 M ethylene glycol; VS3: 3.4 M glycerol + 4.6 M ethylene glycol) before loading into an open-pulled straw (OPS) and plunging directly into liquid nitrogen for storage. At warming, each individual embryo was sequentially placed through warming solutions (WS1: 0.5 M galactose at 37°C; WS2: 0.25 M galactose at 20°C), and warmed embryos were incubated at 37°C in 5% CO2 in humidified air for 20–22 h in 1 ml Syngro® holding medium supplemented with 10% (v/v) alpaca serum to perform an initial in vitro assessment of post-warming viability. Embryos whose diameter increased during culture (n = 2) were transferred individually into synchronous recipients, whereas embryos that did not grow (n = 2) were transferred together into a single recipient to perform an in vivo assessment of post-warming viability. Initial pregnancy detection was performed ultrasonographically 29 days post-transfer when fetal heartbeat could be detected, and one of three recipients was pregnant (25% embryo survival rate). On November 13, 2019, the one pregnant recipient delivered what is believed to be the world's first cria produced from a vitrified-warmed alpaca embryo.


2016 ◽  
Vol 106 (3) ◽  
pp. e187
Author(s):  
M. Pavone ◽  
J.M. Kelsh ◽  
S. Malpani ◽  
R. Confino ◽  
S. Jasti ◽  
...  

2018 ◽  
Vol 30 (1) ◽  
pp. 18 ◽  
Author(s):  
Martyna Heiligentag ◽  
Ursula Eichenlaub-Ritter

The formation of high-quality oocytes depends on complex stage-specific interactions between the germ cell and the somatic compartment involving endocrine, paracrine, and autocrine regulation. Cooperativity in bidirectional signalling and metabolism in response to factors in the microenvironment drive growth, proliferation, cell cycle regulation, spindle formation and the establishment of epigenetic marks in oocytes. This is essential to ensure faithful chromosome segregation and to achieve high oocyte quality, with far-reaching consequences for embryo survival, development and the health of offspring. Oocytes reach developmental capacity throughout early meiotic stages up to full growth and acquisition of nuclear and cytoplasmic maturational competence during folliculogenesis. Improved preantral follicle culture in which ideally intimate contacts between oocyte and somatic cells are retained provides unique opportunities to assess the effects of microenvironment, growth factors, hormones, cryopreservation and environmental exposure on folliculogenesis and oocyte quality. An optimised follicle culture can contribute to the generation of high-quality oocytes for use in fertility preservation in cancer patients, animal breeding and the preservation of endangered species. The past decade has brought about major advances in follicle culture from different species. Recent advances in preantral follicle culture are discussed to assess the effects of environment, adverse exposure, cryopreservation and age on oocyte quality.


2014 ◽  
Vol 26 (1) ◽  
pp. 115 ◽  
Author(s):  
A. F. González-Serrano ◽  
C. R. Ferreira ◽  
V. Pirro ◽  
J. Heinzmann ◽  
K.-G. Hadeler ◽  
...  

Information on how supplementation of high-yield dairy cows with rumen-protected fat affects fertility in cattle herds is scarce. Here, Holstein-Friesian heifers (n = 84) received a supplement consisting of either rumen-protected conjugated linoleic acid (CLA; cis-9,trans-11-CLA and trans-10,cis-12-CLA) or stearic acid 18 : 0 (SA) on top of an isocaloric grass silage diet. Two supplementation doses were used (100 and 200 g d–1). Blood and follicular fluid were collected at the start and end of the supplementation period for analysis of cholesterol, insulin-like growth factor (IGF), and nonesterified fatty acids (NEFA), and for fatty acid profiling. Although cholesterol, IGF, and NEFA levels did not differ among experimental groups, lipid profiles in blood and follicular fluid were affected in a dose-dependent manner by both supplements. After 45 days of supplementation, oocytes were collected by ovum pick-up (OPU). The mRNA relative abundance of target genes (IGF1r, GJA1, FASN, SREBP1, and SCAP) was analysed in single in vitro- (24 h IVM) and in vivo-matured (collected by OPU 20 h after GnRH injection) oocytes and in vitro-produced blastocysts (Day 8) by qPCR (n = 6/group). Lipid profiling of individual oocytes from the CLA-supplemented (n = 37) and the SA-supplemented (n = 50) was performed by desorption electrospray ionization mass spectrometry (DESI-MS). Oocytes from the CLA-supplemented (n = 413) and the SA-supplemented (n = 350) groups were used for assessing maturation and blastocysts development rates. In immature oocytes, CLA supplementation led to an increase of triacylglycerol 52 : 3 [TAG (52 : 3)] and TAG (52 : 2), squalene, palmitic acid 16 : 0, and oleic acid 18 : 1, and decreased abundance of TAG (56 : 3), TAG (50 : 2) and TAG (48 : 1). In vitro-matured oocytes showed different lipid profiles, with increased abundances of TAG (52 : 3), and TAG (52 : 2) as well as phosphatidylinositol 34 : 1 [Plo (34 : 1)], whereas phosphatidylglycerol (34 : 1) [PG (34 : 1)] and palmitic acid 16 : 0 were less abundant in in vitro-matured oocytes. SCAP was significantly down-regulated in in vitro-matured oocytes from supplemented heifers compared with their in vivo-matured counterparts. Maturation (CLA = 74% v. SA = 67%) and blastocyst rates (CLA = 22.4% v. SA = 12.7%) were different among experimental groups. One-way ANOVA and the Tukey-Kramer test were applied for a multiple comparison of means (P-value ≤ 0.05 was considered as statistically significant). In conclusion, we demonstrate here that fatty acid monitoring along different compartments (i.e. blood system, follicular fluid, and intra-oocyte) after rumen-protected fat supplementation of dairy heifer diet reveals nutritional footprints on oocyte quality and embryo development. These results demonstrate the close relationship between nutrition and cattle herd's fertility and, at the same time, support the role of the bovine model for understanding nutritional-dependent fertility impairments.


2021 ◽  
Vol 33 (2) ◽  
pp. 126
Author(s):  
M. Ritz ◽  
A. Gonzalez ◽  
A.-S. Fries ◽  
T. Scheu ◽  
N. Blad-Stahl ◽  
...  

Supplementation of rumen-protected amino acids (RPAA) has proven to be an effective tool to supply limiting AA in dairy diets. Methionine and lysine are the two most limiting AA for lactating dairy cows. Recently, it has been shown that methionine supplementation seems to affect pre-implantation embryos collected from superovulated cows enhancing their developmental competence because there is strong evidence that endogenous lipid reserves serve as an energy substrate (Acosta et al. 2016 Theriogenology 85, 1669–1679). Moreover, higher concentrations of methionine were determined in the follicular fluid of the first dominant follicle postpartum in cows supplemented with rumen-protected methionine and rumen-protected choline from 21 days before calving to 30 days postpartum and it was assumed that higher methionine concentrations in the follicular fluid could affect oocyte quality (Acosta et al. 2017 Theriogenology 96, 1–9). There is no information available so far regarding the effect of a combined methionine and lysine supplementation (each rumen-protected) on oocyte quality. Therefore, the objective of this study was to evaluate the effect of a combined methionine and lysine supplementation during early to mid-lactation on the developmental competence of oocytes collected from lactating dairy cows (days 0 to 100 p.p.). Thirty pregnant multiparous German Holstein dairy cows were grouped 3 weeks before their expected calving date, receiving identical diets. After calving, they were randomly allocated to 2 groups fed a total mixed ration supplemented with (N=14 cows; RPAA) or without (N=16 cows; CON) LysiGEMTM (encapsulated lysine; Kemin Industries) and Metasmart DryTM (isopropyl ester of the hydroxylated analogue of methionine adsorbed onto a silicon dioxide carrier; Adisseo). Starting from 45 days p.p., animals from both groups were submitted to an ovum pickup (OPU) session once a week for at least 8 weeks. Collected cumulus–oocyte complexes (COC) were subjected to a standard invitro production (IVP) protocol (Stinshoff et al. 2014 Reprod. Fertil. Dev. 26, 502–10) including IVM, IVF, and invitro culture (IVC). Cleavage and developmental rates up to the morula/blastocyst stage were recorded on Days 3, 7, and 8. In total, 1211 follicles have been aspirated from RPAA animals compared with 1413 from CON animals, from which 742 and 885 COC were collected, respectively. The calculated recovery rate based on the number of aspirated follicles and collected COC was similar for both groups (61.3±29.4% vs. 62.6±33.5%). Cleavage and developmental rates based on 240 (RPAA group) and 299 (CON group) COC also showed similar results [RPAA: 84.1±5.9% (202/240), 18.3±4.4% (44/240), 18.8±4.7% (45/240); CON: 81.9±8.6% (245/299), 15.4±8.9% (46/299), 16.7±8.4% (50/299)]. In conclusion, supplementation of RPAA (methionine and lysine) had no beneficial effect on the developmental competence of COC obtained from these animals compared with those collected from cows fed the diet without RPAA supplementation.


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