In vivo ultrasound guided transvaginal oocyte collection in the cow

This paper describes the results of the in vivo ultrasound guided follicular aspiration for ovum pick υρ (OPU) in the cow. Twelve non pregnant dry cows aged 4-6 years were used in this experiment. Eight cows underwent OPU during three successive oestrous cycles and another four cows were used as controls having only transvaginal ultrasound scanning of their ovaries. Oocyte collection took place three times during the luteal phase of each natural oestrous cycle (days 3-4,9-11 and 14-17). The content of 326 follicles with a diameter of 4-15mm was aspirated and 104 oocytes were collected (recovery rate 31.9% or 1.55 oocytes per cow and session). The oocyte recovery rate increased after the first three sessions (from 13.04% to 35.0%) and reached levels of υρ to 52.6%. More follicles were aspirated on days 9-11 (133 follicles 40.8%) compared to 111 (34%) follicles on days 14-17 and 82 (25%) on days 3-4) (P<0.05). The evaluation of the collected oocytes revealed that 60 oocytes (57.7%) were suitable for further in vitro manipulation. Neither the origin of the oocyte (left or right ovary) nor the stage of the oestrous cycle affected the recovery rate or the quality of the collected oocytes. There was no significant difference either in the length of the oestrous cycle between the experimental animals and the controls (21.6± 1.4 vs. 22.37±1.0 respectively), or in plasma progesterone concentration in daily collected blood samples from the animals of the two groups. The results of this study are compared to those from the international literature and to the results from endoscopical methods for oocyte recovery. The feasibility of application of this technique to projects designed to improve the genetic merit of cows is discussed.

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In vitro embryo production from bovine oocyte donors aspirated at different frequencies or following treatment with FSH

Proceedings of the British Society of Animal Science ◽

10.1017/s1752756200592680 ◽

1996 ◽

Vol 1996 ◽

pp. 68-68

Author(s):

K.L. Goodhand ◽

R.G. Watt ◽

M.E. Staines ◽

L.C. Higgins ◽

P.J. Broadbent ◽

...

Keyword(s):

Transvaginal Ultrasound ◽

Genetic Change ◽

Bovine Oocyte ◽

Ultrasound Guided ◽

Embryo Production ◽

Follicular Aspiration ◽

Oocyte Donors ◽

Pre Treatment

The combination of in vivo recovery of oocytes using transvaginal ultrasound guided aspiration and subsequent in vitro embryo production can be used to increase the rate of genetic change for efficiency of beef production by increasing selection intensity and reducing generation interval. The total number of oocytes recovered by aspiration and embryos produced is directly proportional to the number of aspiration sessions whether recovery takes place once or twice weekly. Pre-treatment of oocyte donors with FSH has been shown to improve the number of follicles available for aspiration but effects on embryo production have been conflicting (Bungartz et al., 1995; Goodhand et al., in press). The objective of this experiment was to compare the effect on embryo production of frequency of follicular aspiration and pre-treatment of donor cattle with FSH.

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In vitro embryo production from bovine oocyte donors aspirated at different frequencies or following treatment with FSH

Proceedings of the British Society of Animal Science ◽

10.1017/s0308229600030397 ◽

1996 ◽

Vol 1996 ◽

pp. 68-68

Author(s):

K.L. Goodhand ◽

R.G. Watt ◽

M.E. Staines ◽

L.C. Higgins ◽

P.J. Broadbent ◽

...

Keyword(s):

Transvaginal Ultrasound ◽

Genetic Change ◽

Bovine Oocyte ◽

Ultrasound Guided ◽

Embryo Production ◽

Follicular Aspiration ◽

Oocyte Donors ◽

Pre Treatment

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Effect of administering a crude equine gonadotrophin preparation to mares on follicular development, oocyte recovery rate and oocyte maturation in vivo

Reproduction ◽

10.1530/reprod/118.2.351 ◽

2000 ◽

pp. 351-360 ◽

Cited By ~ 15

Author(s):

I Bruck ◽

J Bezard ◽

M Baltsen ◽

B Synnestvedt ◽

I Couty ◽

...

Keyword(s):

Oocyte Maturation ◽

Recovery Rate ◽

In Vitro Maturation ◽

Follicular Development ◽

Vitro Fertilization ◽

Nuclear Maturation ◽

Oocyte Yield ◽

Oocyte Recovery

In mares, the shortage of oocytes and the variability in nuclear maturation at a certain time of the oestrous cycle hinders the optimization of methods for in vitro maturation and in vitro fertilization. Increasing the number of small-to-medium-sized follicles available for aspiration in vivo may increase the overall oocyte yield. The aims of the present study were to investigate whether administration of crude equine gonadotrophins affects follicular development, oocyte recovery rate, in vivo oocyte maturation and follicular concentrations of meiosis-activating sterols. During oestrus, all follicles >/= 4 mm were aspirated from 19 pony mares (first aspiration: A1). Over the next 8 days, the mares were treated daily with either 25 mg crude equine gonadotrophins (n = 10) or physiological saline (n = 9). Between day 1 and day 8, follicular growth was monitored by ultrasonography. On day 8, all follicles >/= 4 mm were evacuated (second aspiration: A2) and nuclear maturation of the recovered oocytes was assessed after orcein staining. Follicular growth between A1 and A2, as well as the number and size of follicles at A2 were similar for control mares and mares treated with crude equine gonadotrophins. The oocyte recovery rates at A1 and A2 were similar. At A2, the oocyte recovery rate and oocyte maturation in vivo were not affected by treatment with crude equine gonadotrophins. The number of expanded cumulus oophorus complexes recovered from follicles </= 29 mm was significantly higher at A1 than at A2. The number of oocytes at the germinal vesicle stage was significantly higher at A2 (41.5%) than at A1 (17.8%). Meiosis-activating sterols (FF-MAS and T-MAS) were identified in follicular fluid recovered at A2. Follicular concentrations of FF-MAS and T-MAS were unaffected by treatment with crude equine gonadotrophins. The present study demonstrates that follicular aspiration during oestrus allowed a new follicular population to develop and resulted in a higher degree of synchronization of oocyte development with respect to cumulus expansion and nuclear maturation. The availability of a more homogeneous population of oocytes might facilitate a better optimization of in vitro maturation and in vitro fertilization techniques in mares. Administration of crude equine gonadotrophins during early dioestrus did not affect the growth of small follicles, the oocyte yield after aspiration or oocyte maturation in vivo.

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263 IN VITRO FERTILIZATION AND EMBRYO DEVELOPMENT OF CUMULUS - OOCYTE COMPLEXES COLLECTED BY ULTRASOUND-GUIDED FOLLICULAR ASPIRATION IN LLAMAS TREATED WITH GONADOTROPIN

Reproduction Fertility and Development ◽

10.1071/rdv22n1ab263 ◽

2010 ◽

Vol 22 (1) ◽

pp. 288 ◽

Cited By ~ 4

Author(s):

M. A. Berland ◽

A. von Baer ◽

V. Parraguez ◽

P. Morales ◽

G. P. Adams ◽

...

Keyword(s):

In Vitro Fertilization ◽

Embryo Development ◽

Animal Health ◽

Ovarian Response ◽

Ultrasound Guided ◽

Vitro Fertilization ◽

Follicular Aspiration ◽

Oocyte Collection ◽

Health Canada

We have previously documented that both FSH and eCG are equally effective in inducing ovarian superstimulation in llamas, resulting in the recovery of a high number of expanded COC suitable for in vitro fertilization (Ratto et al. 2005 Theriogenology 63, 2445-2457). The objective of the study was to evaluate the ovarian response, morphology, and competence of COC collected by ultrasound-guided follicular aspiration in llamas treated with FSH or eCG. Llamas were assigned randomly into 2 groups (n = 16 per group) and treated for 48 h after follicle ablation with (1)25 mg of FSH (Folltropin, Bioniche Animal Health Canada Inc., Belleville, Canada) i.m. twice daily for 4 d; or (2) 1000IU of eCG (Novormon, Bioniche Animal Health Canada) as a single i.m. dose. The starting of gonadotropin treatment was considered Day 0. Both groups were given an i.m. dose of 5 mg of Armour Standard LH (Lutropin, Bioniche Animal Health Canada) on Day 6, and COC were collected by transvaginal ultrasound follicle aspiration of all follicles ≥7 mm on Day 7. The ovarian response was assessed by transrectal ultrasonography using a 7.5-MHz linear-array transducer (Aloka SSD-500, Clinics, Santiago, Chile) immediately before oocyte collection at 24 to 26 h after LH treatment in both groups. The COC were classified as expanded, compact, denuded, or degenerated. Expanded COC collected from FSH- (n = 147) and eCG-treated llamas (n = 141) were fertilized in vitro using epididymal sperm as previously described (Ratto et al. 2006 Anim. Reprod. Sci. 97, 246-257). Gametes were co-incubated at 38.5°C in air with 5% CO2 and high humidity for 18 h. After in vitro fertilization, presumptive zygotes were co-culture in SOF medium supplemented with 0.6% of BSA with llama granulosa cells at 39°C, 5% CO2, 5% O2, and 90% N2 for 7 days. Embryo development was evaluated on Days 2, 5, and 7 of in vitro culture (Day 0 = IVF). Data were analyzed by Student’s t-test or Fisher’s exact test and presented as mean ± SEM. The FSH and eCG treatment groups did not differ with respect to the number of follicles ≥7 mm at the time of COC collection (16.0 ± 2.7 v. 14.0 ± 1.9; P = 0.5), the number of COC collected (11.5 ± 1.9 v. 9.7 ± 1.2; P = 0.4), or the collection rate per follicle aspirated (77.0 v. 71.5%; P = 0.2). No difference was detected between FSH and eCG-treated llamas in the number of expanded COCs (9.8 ± 1.4 v. 9.4 ± 1.2; P = 0.8). The percentage of presumptive zygotes to develop into 2 to 8 cells on Day 2 (65.3 v. 63.1), morulas on Day 5 (46.2 v. 42.5), and blastocyst stage on Day 7 (23.1 v. 20.5) did not differ (P > 0.05) between FSH and eCG-treated llamas, respectively. In conclusion, FSH and eCG treatments were equally effective for ovarian superstimulation and oocyte collection. The recovery of a high number of expanded COC can be used directly for in vitro fertilization and their competence is not affected by gonadotropin treatment. The study was supported by Convenio Desempeño en Investigacion (2007-DGI-CDA-04), Universidad Catolica de Temuco.

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158 The Effect of Follicular Wave Phase at Time of Ovum Pick-Up on Bovine Oocyte Cytoplasmic Maturation and Developmental Competence

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab158 ◽

2018 ◽

Vol 30 (1) ◽

pp. 218

Author(s):

B. A. Foster ◽

F. A. Diaz ◽

E. J. Gutierrez ◽

K. R. Bondioli

Keyword(s):

Embryo Development ◽

Transvaginal Ultrasound ◽

Oocyte Quality ◽

Developmental Competence ◽

Developmental Potential ◽

Follicular Wave ◽

Wave Phase ◽

Oocyte Collection ◽

Significant Difference

During oocyte collection, follicular wave phase is unknown, although differences in follicle environment may have dramatic effects on oocyte quality. This project was performed to determine whether oocyte collection during different phases of the follicular wave affects oocyte competence. Oocytes were collected via transvaginal ultrasound guided oocyte aspiration from 18 cows, at 4, 8, and 12 days following dominant follicle removal, representing follicle wave emergence, peak, and atresia, respectively (160, 314, and 273 oocytes, respectively). Once recovered, oocytes were graded and assigned to either being held as immature or matured in vitro for 24 h. Oocytes were then stained in Mitotracker deep red, fixed and stained with an anti-IP3R1 primary antibody and an Alexa Fluor 488-conjugated secondary antibody, before being stained with DAPI, to identify mitochondria, inositol triphosphate receptor 1 (IP3R1), and chromatin respectively. Mitochondria were analysed based on cytoplasmic distribution and classified as peripheral (immature), diffuse, central (mature), or sparse. Expression of IP3R1 was measured as corrected total cell fluorescence in Image J (National Institutes of Health, Bethesda, MD, USA). Staining patterns were analysed using ANOVA. A subset of the matured oocytes was stained with Fluo-3 to measure cytoplasmic calcium levels. These oocytes were then parthenogenetically activated before being imaged again to view changes in calcium levels, and presumptive embryos were cultured for 4 days. Fluo staining was measured as intensity levels (none, slight, moderate, high) and differences in development and stain levels were analysed using the Kruskal-Wallis test. Although mitochondria location was unaffected by collection day, it was significantly affected by maturation status (P = 0.0036). However, oocytes showed incomplete mitochondrial maturation, with mitochondria residing in the diffuse orientation in the majority of oocytes. Expression of IP3R1 appeared to be more sensitive to treatment. Expression significantly increased as meiosis proceeded (P = 0.0081) and there was a significant difference in expression between oocyte collection days (P = 0.0026). The interaction between collection day and maturation status also had a significant effect (P = 0.048), with mature oocytes showing an increase in IP3R1 expression, most notable in those collected on Day 4. Oocyte quality had a notable effect on the ability of oocytes to progress through meiosis (P = 0.054) and on mitochondrial location (P = 0.053), with AB oocytes showing better maturation parameters in both respects. Although the day of collection did not affect embryo development, Fluo stain intensity was an indicator of embryo developmental potential (P = 0.053), with oocytes having decreased potential to develop if the initial calcium levels were moderate to high. Results suggest that oocyte collection during wave emergence yields a slight advantage in oocyte quality. Although IP3R1, necessary for Ca2+ spikes during fertilization, indicates competence, high levels of cytoplasmic Ca2+ at the time of activation appear to be detrimental to embryo development.

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In Vitro Embryo Production and Transfer of Bubaline Embryos using Oocytes Derived from Transvaginal Ultrasound-Guided Follicular Aspiration (TUFA)

International Journal of Applied Sciences and Biotechnology ◽

10.3126/ijasbt.v2i2.10369 ◽

2014 ◽

Vol 2 (2) ◽

pp. 180-184

Author(s):

FP Aquino ◽

Eufrocina P. Atabay

Keyword(s):

Transvaginal Ultrasound ◽

Blastocyst Stage ◽

Ultrasound Guided ◽

Blastocyst Development ◽

Embryo Production ◽

Follicular Aspiration ◽

Group 2 ◽

Group 1 ◽

Donor Cows

Transvaginal ultrasound-guided follicular aspiration (TUFA) has become a popular tool for embryo production in vitro due to its high degree of repeatability in terms of recovering oocytes from live animals. In Study 1, the quantity and quality of oocytes from Bulgarian Murrah buffalo cows (n=10) of varying ages (Group 1, 8-12; and Group 2, 13-17 years) were assessed. Group 1 buffalo donor cows yielded significantly higher (P<0.05) number of oocytes vs Group 2 buffalo donor cows (71 vs 29 oocytes, respectively), though in terms of oocyte quality, no difference was observed. In Study 2, oocytes collected (n=100) in Study 1 were matured, fertilized in vitro and the resulting zygotes were cultured which developed to blastocyst stage embryos. The maturation, fertilization and blastocyst development rates obtained were 53.0%, 40.0% and 32.5%, respectively. In Study 3, the viability of resulting blastocyst stage embryos was determined by transferring to recipient cows. Of 10 recipients 1 got pregnant and delivered a 35 kg male calf after 310 days gestation period. Overall, the results of the studies conducted demonstrated the potential of TUFA technology in the in vitro production of embryos which eventually could be used in the production of live offspring.DOI: http://dx.doi.org/10.3126/ijasbt.v2i2.10369Int J Appl Sci Biotechnol, Vol. 2(2): 180-184

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The Effectiveness of Candidate Probiotic Bacteria to Control Vibriosis Disease

Aquatic Science and Technology ◽

10.5296/ast.v5i2.11594 ◽

2017 ◽

Vol 5 (2) ◽

pp. 1

Author(s):

Mulyati Mulyati ◽

Suryati Suryati ◽

Irfani Baga

Keyword(s):

Survival Rate ◽

Sea Water ◽

Challenge Test ◽

Probiotic Bacteria ◽

Pathogenicity Test ◽

Inhibitory Ability ◽

Significant Difference ◽

Portunid Crab

The study aims to isolate, characterize, and examine probiotic bacteria's inhibitory ability against Vibrio harveyi bacteria, both in-vitro and in vivo. Methods used in the study consist of 1) An Isolation of Candidate Probiotic Bacteria, 2) An Antagonistic Test of Candidate Probiotic Bacteria in vitro, 3) An Identification of Bacteria, 4) A Pathogenicity Test of Candidate Probiotic Bacteria, 5) An Antagonistic Test of Candidate Probiotic Bacteria against V. harveyi in vivo. According to the isolation of candidate probiotic bacteria, there are 18 isolated candidate probiotic. After being tested for its inhibitory ability in vitro, there are 8 isolates with zone of inhibition as follows: isolate MM 7 from intestine (22 mm), isolate MM 6 from intestine (12 mm), isolate MM 10 from sea water (10 mm), isolate MM 5 from intestine (9 mm), isolate MM 4 from intestine (8 mm), isolate MM 3 from intestine (7 mm), isolate MM 2.2 from intestine (7 mm), isolate MM 2.1 from intestine (7 mm). Eight genera of the candidate probiotic bacteria is derived from Portunid crab, they are Staphylococcus, Streptococcus, bacillus, vibrio, Alcaligenes, Lactobacillus, micrococcus. Before proceeding the V. harveyi bacterial challenge test in vivo, three potential isolates consisting of MM6, MM7 and MM10 as the probiotic bacteria are pathogenicity-tested against V. harveyi. The survival rate of Portunid crab on pathogenicity test using MM6, MM7 and MM10 generates 91.11-100%, while the control generates 100% survival rate. Variance analysis result through post-hoc Tukey's Honest Significant Difference (HSD) test at 95% confidence interval indicates that isolate MM7 and MM10 are significantly able to increase hatchling Portunid crab's survival rate.

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The macaque model for in vitro fertilization: Superovulation techniques and ultrasound‐guided follicular aspiration

Journal of Medical Primatology ◽

10.1111/j.1600-0684.1991.tb00501.x ◽

1991 ◽

Vol 20 (3) ◽

pp. 110-116 ◽

Cited By ~ 1

Author(s):

Catherine A. VandeVoort ◽

Alice F. Tarantal

Keyword(s):

In Vitro Fertilization ◽

Ultrasound Guided ◽

Vitro Fertilization ◽

Macaque Model ◽

Follicular Aspiration

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Multimodal Diagnostics of Microrheologic Alterations in Blood of Coronary Heart Disease and Diabetic Patients

Diagnostics ◽

10.3390/diagnostics11010076 ◽

2021 ◽

Vol 11 (1) ◽

pp. 76

Author(s):

Anastasia Maslianitsyna ◽

Petr Ermolinskiy ◽

Andrei Lugovtsov ◽

Alexandra Pigurenko ◽

Maria Sasonko ◽

...

Keyword(s):

Coronary Heart Disease ◽

Heart Disease ◽

Red Blood Cells ◽

Blood Cells ◽

Optical Methods ◽

Diabetic Patients ◽

Aggregation Index ◽

Significant Difference

Coronary heart disease (CHD) has serious implications for human health and needs to be diagnosed as early as possible. In this article in vivo and in vitro optical methods are used to study blood properties related to the aggregation of red blood cells in patients with CHD and comorbidities such as type 2 diabetes mellitus (T2DM). The results show not only a significant difference of the aggregation in patients compared to healthy people, but also a correspondence between in vivo and in vitro parameters. Red blood cells aggregate in CHD patients faster and more numerously; in particular the aggregation index increases by 20 ± 7%. The presence of T2DM also significantly elevates aggregation in CHD patients. This work demonstrates multimodal diagnostics and monitoring of patients with socially significant pathologies.

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Effects of Surface Protein Adsorption on the Distribution and Retention of Intratumorally Administered Gold Nanoparticles

Pharmaceutics ◽

10.3390/pharmaceutics13020216 ◽

2021 ◽

Vol 13 (2) ◽

pp. 216

Author(s):

Rossana Terracciano ◽

Aobo Zhang ◽

E. Brian Butler ◽

Danilo Demarchi ◽

Jason H. Hafner ◽

...

Keyword(s):

Treatment Modalities ◽

Emission Spectrometry ◽

Limiting Factor ◽

High Resolution Computed Tomography ◽

Quantitative Ct ◽

Heterogeneous Distribution ◽

Significant Difference ◽

Intratumoral Distribution

The heterogeneous distribution of delivery or treatment modalities within the tumor mass is a crucial limiting factor for a vast range of theranostic applications. Understanding the interactions between a nanomaterial and the tumor microenvironment will help to overcome challenges associated with tumor heterogeneity, as well as the clinical translation of nanotheranostic materials. This study aims to evaluate the influence of protein surface adsorption on gold nanoparticle (GNP) biodistribution using high-resolution computed tomography (CT) preclinical imaging in C57BL/6 mice harboring Lewis lung carcinoma (LLC) tumors. LLC provides a valuable model for study due to its highly heterogenous nature, which makes drug delivery to the tumor challenging. By controlling the adsorption of proteins on the GNP surface, we hypothesize that we can influence the intratumoral distribution pattern and particle retention. We performed an in vitro study to evaluate the uptake of GNPs by LLC cells and an in vivo study to assess and quantify the GNP biodistribution by injecting concentrated GNPs citrate-stabilized or passivated with bovine serum albumin (BSA) intratumorally into LLC solid tumors. Quantitative CT and inductively coupled plasma optical emission spectrometry (ICP-OES) results both confirm the presence of particles in the tumor 9 days post-injection (n = 8 mice/group). A significant difference is highlighted between citrate-GNP and BSA-GNP groups (** p < 0.005, Tukey’s multiple comparisons test), confirming that the protein corona of GNPs modifies intratumoral distribution and retention of the particles. In conclusion, our investigations show that the surface passivation of GNPs influences the mechanism of cellular uptake and intratumoral distribution in vivo, highlighting the spatial heterogeneity of the solid tumor.

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