Visit to Austria

On 23-24th June Gergő and Ákos went to Austria to visit Dr. Franz Lahnsteiner and Florian A. Kunz at the Bundesamt für Wasserwirtschaft in Scharfling and its fish farm in Kreuzstein. In addition to being partners in the COST Action FA1205 AQUAGAMETE, we have a long history of collaboration with Franz and hist colleagues and we share a common interest in the culture of various fish species including salmonids, percids, tench and burbot. We discussed the possiblities of a joint Austrian-Hungarian project on various aspects of fish sperm cryopreservation.

Dr. Franz Lahnsteiner, Florian A. Kunz and Ákos discussing project possibilites in Scharfling, Austria. The beer is alcohol-free of course 😉. Photo by Gergő

6th COST AQUAGAMETE Training School in Rennes

Ágnes was participated as a selected candidate in 6th COST AQUAGAMETE training school in Rennes, France. It was focused on the Molecular Basis of Gamete Quality and Reproduction, with a strong orientation towards genomic tools. The course was organized by the Fish Physiology and Genomics Department of INRA between June 6th and 10th 2016, and coordinated by Julien Bobe and and Catherine Labbé.

The course included conferences on:

• Sperm genetic damage and repair in Fish (Paz Herraez, Spain)
• Overview of sequencing technologies and strategies available to analyze fish reproduction (Yann Guiguen, France)
• Epigenetics and inheritance (Francesc Piferrer, Spain)
• Transgenesis approaches in fish reproduction  
• CrispR/Cas9 technology for research on fish reproduction (Amaury Herpin, France)
• microRNA and female fecundity (Amine Bouchareb, France)

The practical sessions included experiments on:

• RNA extraction and meausing of quality and quantity
• Microarray analysis of fish samples in reproduction researc
• Sequence searching including tools presentation, primer design, phylogeny/syntheny search

 

Cover the microarray for analysis

Everybody paid attention to the analysis (except Carina and Effrossyni:P)

The group

Graduation ceremony

On Friday, June 17th, Gergő has officially received his PhD degree and Ákos his title as Dr. habil. at the gaduation ceremony of Szent István University.

A very official photo of the awardees

PhD defence of Gergő

Today Gergely Bernáth from our group has succesfully defended his PhD entitled "Development of fish sperm qualification systems for economic purposes". 

The committee evaluated Gergő's work with outstanding credit points.

Another new paper accepted

A new paper by our group with the first authorship of Gergely Bernáth was accepted for publication in Animal Reproduction Science. For more information, please consult this link: http://www.sciencedirect.com/science/article/pii/S0378432016302226

Commercial-scale out-of-season cryopreservation of Eurasian perch (Perca fluviatilis) sperm and its application for fertilization

Abstract
The quality and fertilizing capacity of perch (Perca fluviatilis) sperm collected outside of the spawning season (off-season) and cryopreserved at a commercial scale, were tested. Basic parameters (equilibration time, dilution ratio, sperm concentration, post-thaw motility duration) which can have a significant effect on cryopreservation success were systematically investigated for effects on sperm quality using computer assisted sperm analysis (CASA). No significant decrease in progressive motility (pMOT) and straightness (STR) of fresh-diluted sperm was recorded among groups equilibrated for 0, 30 or 60 minutes in an extender with cryoprotectants. Curvilinear velocity (VCL) was reduced significantly after 30 minutes (30 min: 146 ± 15 μm/s, 60 min: 124 ± 18 μm/s) of equilibration compared to the control (174 ± 9 μm/s). After thawing, no decrease in pMOT or VCL was observed at different equilibration times in any of the analyzed groups. No correlation was observed among progressive motility, dilution ratios (p = 0.7) and cell concentrations (p = 0.1). The use of different activating solutions resulted in similar pMOT and VCL in the first 120 seconds post-thaw. Nevertheless, post-thaw sperm motility was reduced after 30 seconds using all activators. Motility parameters with low variation were recorded after thawing of 57 straws (pMOT: 37 ± 7%, VCL: 92 ± 10 μm/s, STR: 89 ± 3%). Ten randomly selected straws from commercial-scale cryopreservation resulted in a high fertilization rate (cryopreserved sperm: 72 ± 14%, fresh control: 94 ± 2%). An optimized commercial-scale cryopreservation protocol was successfully developed for Eurasian perch. The applicability of the off-season collected perch sperm for cryopreservation and fertilization was demonstrated.

New paper accepted in General and Comparative Endocrinology

A new paper by our group was accepted for publication in General and Comparative Endocrinology as a part of the proceedings of last year's 5th International Workshop on the Biology of Fish Gametes held in Ancona, Italy on September 7-11, 2015. For more information please consult this link: http://www.sciencedirect.com/science/article/pii/S0016648016301319

DEVELOPMENT OF SPERM VITRIFICATION PROTOCOLS FOR FRESHWATER FISH (EURASIAN PERCH, Perca fluviatilis) AND MARINE FISH (EUROPEAN EEL, Anguilla anguilla)

Abstract
Vitrification was successfully applied to the sperm of two fish species, the freshwater Eurasian perch (Perca fluviatilis) and marine European eel (Anguilla anguilla). Sperm was collected, diluted in species-specific non-activating media and cryoprotectants and vitrified by plunging directly into liquid nitrogen without pre-cooling in its vapor. Progressive motility of fresh and vitrified-thawed sperm was evaluated with computer-assisted sperm analysis (CASA). Additional sperm quality parameters such as sperm head morphometry parameters (in case of European eel) and fertilizing capacity (in case of Eurasian perch) were carried out to test the effectiveness of vitrification. The vitrification method for Eurasian perch sperm resulting the highest post-thaw motility (14 ± 1.6%) was as follows: 1:5 dilution ratio, Tanaka extender, 30% cryoprotectant (15% methanol + 15% propylene-glycol), cooling device: Cryotop, 2 μl droplets, and for European eel sperm: dilution ratio 1:1, with 40% cryoprotectant (20% MeOH and 20% PG), and 10% FBS, cooling device: Cryotop, with 2 μl of sperm suspension. Viable embryos were produced by fertilization with vitrified Eurasian perch sperm (neurulation: 2.54±1.67%). According to the ASMA analysis, no significant decrease in head area and perimeter of vitrified European eel spermatozoa were found when compared to fresh spermatozoa.

Summer Course in Aquaculture

The Department of Aquaculture of Szent István University is organizing a CEEPUS Summer Course in Aquaculture on July 07-09th, 2016. 

For more information, please consult this link: http://mkk.szie.hu/dep/halt/ceepus_summer_course_2016/

II Training School on Epigenetics in Reproductive Biology

Jelena and Zoran participated in this training school which was organized by COST Action EPICONCEPT (FA1201) at Faculty of Veterinary Sciences, University of Murcia, Murcia, Spain.

Some of the lecturers were renowned scientists in this field from UK, Belgium, Netherlands and Spain and among participants were young scientists from 20 countries. 

Faculty of Veterinary Sciences and Veterinary hospital

part of the schedule

lab work

lab work, data analysis

between sessions

between sessions

with our new friends and colleagues from Netherlands, Poland, Croatia and Macedonia

Field work in Slovenia

Five of us were in Slovenia for sperm cryopreservation work from 11-04-2016 to 15-04-2016.

our team: Gergő, Ákos, Kinga, Eszti and Zoli

Our team was complemented with the company of a Portuguese collegue, Carina Caldeira, who is an IMPRESS student at the company Proiser. She is currently on a Short-term Scientific Mission of the COST Action FA-1205 AQUAGAMETE carrying out sperm analysis of various fish and shellfish species.

Carina and the Slovenian hills

Sperm samples from grayling (Thymallus thymallus) individuals were taken in the area of the Angling Association of Tolmin, with the help of Dušan Jesenšek and his team.

fish collection

grayling and marble trout collected from the river

ready for sampling

sperm cryopreservation in the field

sperm samples of wild-caught grayling were stored for future fertilizations

 We also worked with the grayling of the fish farm:

sperm stripping

  

Carina was working on the improvement of the PROISER CASA system, Eszti was doing vitrification experiments, and the rest of the team did cryopreservatin trials.

fertilization tests were also carried out with cryopreserved and vitrified sperm samples

Transplantation work in Slovenia part 2

Jelena and Zoran were in Slovenia at the Department for Animal Science and with Slovenian colleagues did germ cell transplantation experiments on trout species. 

 

trout larvae

happy face with trout larvae

preparing larvae for microinjection

transplantation by Zoran

transplantation by Jelena

post-transplantation face

post-transplantation face

bye, bye babies, see you soon

Habilitation lecture of Ákos

On Thursday 24th March Ákos had his habilitation lecture: he made presentations both in English and Hungarian languages. Ákos received the maximal number of points from the scientific commitee.

during the English presentation

questions and comments

Transplantation of spermatogonia in Japan

Two members of our group (Jelena and Zoran) spent one month in Japan working in the labs of prof. dr Goro Yoshizaki (Tokyo University of Marine Science and Technology). The topic of their stay was training in transplantation of spermatogonia on trout and zebrafish.

One week they spent at the trout farm in Oizumi region working on transplantation of sprematogonia.  

With part of the Oizumi team

Preparing the tissue

The needle is ready

Preparing the larve

And start transplantation

Small sightseeing of Oizumi region

Three weeks they spent in the labs Tokyo working on transplantation of spermatogonia on zebrafish.

We were honored to be there

Zoran and Li san

Happy face in the lab

Preparing the tissue

Transplantation

Transplantation