Institute of Animal Physiology CBs SAS

International collaborations in 2017:

In the ongoing collaboration with the DNA Integrity Laboratory (Institute of Animal Physiology and Genetics of the ASCR, Liběchov, Czech republic) the early embryo response was studied at the subcellular level after DNA damage caused with exogenous inducer. Our results documented an increased degree of DNA damage defined by the number of double-strand breaks on the DNA molecule and increased histone H2AX phosphorylation. It has been shown that immediately after DNA damage, the first mitotic cycle is prolonged but subsequently damaged preimplantation embryo progressively stops its development. Analysis of the embryonic cell phenotype revealed a process of missegregation of mitotic chromosomes during cell division. At lower degree of DNA damage, it was found that signaling of the cell cycle control mechanisms was not activated but this embryo did not complete replication of the inherent genome during the first cycle followed with enter into second cleavage stage with decreased volume of DNA.
Within the COST FA 1302 METHAGENE project, Dr. Grešáková on days 19-21. June 2017 attended an internship in Sweden in Umea (Training school 2017 – Dr. Grešáková). The course entitled “Breeding for complex traits” focused on the study and evaluation of methane emissions in ruminant breeds. In October 2017, a final conference meeting was held in Caserta (Italy). From our Institute Dr. Váradyová, Dr. Kišidayová and Dr. Grešáková were attended. Dr. Grešáková had a lecture entitled “Effect of selected medicinal herbs from Central Europe on methane production from in vitro ruminal fermentations”. With the financial support of the METHAGENE-STMS project, Dr. Monika Holodová took part in a internship (9.10 – 9.11.2017) at the Department of Animal Nutrition and Feed Management, Poznan University of Life Sciences in the laboratory of Prof. Adam Cieslak. She was involved in work to study the nutritional factors affecting methane production in dairy cows with different feed sources and typical Polish diets (STSM 2017- Dr. M. Holodová). During the final meeting in Caserte, the results for the whole project duration from 2013 to 2017 were evaluated. Final meeting was focusing on new breeding programs aimed at reducing methane emissions from ruminants. Throughout the METHAGENE project, staff from our Institute actively participated in experiments related to the large-scale measurement of methane in ruminants and the factors that affect it. We participated in experiments at the Institute of Agricultural Sciences at the Department of Nutrition in Switzerland (ETH Zürich, Switzerland – STSM 2015 – Dr. J. Pisarcikova) at the National Institute for Agricultural Research in France (INRA, Clermont-Ferrand, France – STSM 2016 – L. Kolesar Fecskeová) We actively participated in the international practical course on methane emission measurement techniques, statistical comparisons, manipulation and analysis of acquired data in ruminants (Poznan University of Life Sciences, Department of Genetics and Animal Breeding and Department of Animal Nutrition and Feed Management, Poland – Training schol 2016 – Dr. M. Wencelová). Through its network, the METHAGENE project, created a platform for efforts to develop tools for the genetic reduction of methane emissions in ruminants (Dr. Váradyová).
Within the study stay in Poland (laboratory of Prof. Adam Cieslak, Department of Animal Nutrition and Feed Management, Poznan University of Life Sciences, Poland) with the financial support of project COST FA1302 METHAGENE (09/10/2017 – 09/11/201, STMS-M. Holodová). Dr. Monika Holodová, PhD student was participated on project concern on nutritional factors affecting methane production in dairy cows fed diets with different forage source with aim to examine the effect of two diets on methane production and emission in high-yielding dairy cows.
Magdalena Bryszak, PhD student at the Department of Animal Nutrition and Feed Management, Poznan University of Life Sciences in Poland was attended a 3-month internship (25 August – 25 November, 2017) at the Department of Digestive Tract Physiology (Laboratory of Digestion and Nutrition) of IAP-SAS. The topic of internship was concerned on plant secondary metabolites in ruminant nutrition. The stay was covered by EU funds “(The Project You study – You practice. Internship programme for students of the Faculty of Veterinary Medicine and Animal Sciences of Poznań University of Life Sciences in Poznań – Internship supervisor: Dr. Zora Váradyová)”.
In the framework of the project „European network on the factors affecting the gastro-intestinal microbial balance and the impact on the health status of pigs, COST-FA1401-PiGutNet (Short term missions), Dr. Ivana Kubašová had a study stay at the University of Life Sciences in As (Norway). She purified bacteriocin, antimicrobial substance-enterocin B produced by Enterococcus faecium strain IK25; this Ent can be allotted to class II enterocins; it means termo-stable small peptide. Purification of Ent B gives possibilities for its application in husbandry of food producing and social animals as well (Dr. Kubašová).
In the framework of Institutional agreement between our Institute and The Kielanowski Institute of Animal Physiology and Nutrition, Jablonna, Polish Academy of Sciences, sensitivity of Buttiauxiella spp. was tested (isolated from faeces of roe and red deers in forestry areas of Poland) to enterocins detected and characterized in our workplace. In wildlife animals exist only limited information regarding the microbiota. Faecal samples from 21 wildlife animals were screened (9 red deer and 12 roe deers). Representatives of the genus Buttiauxiella from the Family Enterobacteriacae were identified using Maldi-Tof mass spectrometry and phenotypization; among 21 colonies, the species Buttiauxiella gaviniae, B. ferragutiae, B. agrestis were detected. They were mostly sensitive to antibiotics; however, B. agrestis 2/109/1 strain was resistant to three antibiotics and sensitive to enterocins at inhibition activity from 100 to 25 600 AU/ml. This results contributed to basic research-microbiota of wildlife animals and also to inhibition spectra of enterocins. Buttiauxiella inhibition due to enterocins noone tested previously (Dr. Lauková a kol.).
Moreover, in the framework of this COST FA 1401 project (European network on the factors affecting the gastro-intestinal microbial balance and the impact on the health status of pigs, PiGutNet) was performed the experiment on swine together with the Kielanowski Institute of Physiology and Nutrition of Polish Academy of Sciences in Jablonna near Warsaw (Poland). The aim of the experiment was to check influence of different forms of zinc and fibre in diet on growth parameters, biochemestry, microbiota, mineral and antioxidant staus of swine. Results achieved were presented on the international conferences (Dr. Grešáková, Čobanová).
In the framework of bilateral co-operation with Faculty of Veterinary Medicine, Technical University in Lisbon (Portugal) continues our co-operation in plantaricin gene testing from Lactobacillus plantarum strains isolated from various animals. Nine plantaricin genes were controlled (A, B, J, M, R, C, D, N, K) in 8 L. plantarum strains originated from faeces of monkey, malpa, dog, duck, hen, pheasant, from ruminal content and ewes milk. In L. plantarum L17/1, S14B, L13Ba (ewes milk, hen, pheasant) were detected genes for all plantaricins tested. After in vitro tests of inhibition activity using the most number of indicator strains of various origin we will focus on plantaricin purification from the most active strains with a braod antimicrobial spectrum to obtain substances for application (Dr. Lauková, Dr. Kandričáková).

Some of previous cooperations:

An intensive collaboration continued within the DAAD-SAS exchange programm between Slovakia and Germany 2015-2016 (the project „Hormonal dysregulations in a diabetic pregnancy -Molecular mechanisms of metabolic and stress-related adaptation of mammalian embryos in a diabetic pregnancy“, principal investigator from Slovakia – Dr. Štefan Čikoš, Phd). Two stays were completed in 2016: Dr. Babeľová-Kubandová and Dr. Špirková (Halle, november 2016), and Dr. Navarrete Santos and Dr. Seeling (Košice, august 2016).
The one-week international training school entitled Contemporary feed production-technological and nutritional aspects was organised by the COST Action FA 1401 „European network on the factors affecting the gastro-intestinal microbial balance and the impact on the health status of pigs (PiGutNet)“ in February 2016. Dr. Ľubomíra Grešáková with other foreign young researchers attended the course that took place at the Institute of Food Technology, Research Centre Feed-to-Food, Novi Sad in Serbia. The workshop was focused on the getting the latest knowledge and practice in the field of modern feed technology and production, new trends in the animal nutrition as well as special processing needs for feeding concepts for young animals. The lectures program was enriched with two scientific lessons by guests from prominent universities in Belgium and Italy who have dealt with the problems in the animal nutrition. Theoretical lectures were combined with practical work and laboratory testing of feed materials. The stay at the course established good contacts and the international cooperation.
In the frame of the cooperation with the Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences in Jablonna, in Poland we carried out the common experiment in October 2016. The main objectives of the experiment are to compare and evaluate the effect or interaction of two different dietary sources of fibre and zinc on mineral and antioxidant status, and also the health and performance of the piglets after weaning. (Dr. Čobanová, Dr. Grešáková).
Regarding the collaboration of our Institute (Laboratory of Animal Microbiology) with Kielanowski Institute of Animal Physiology and Nutrition, Polish Academy of Sciences, Jablonna near Warsaw (Poland), microbial profile of horses derived from different regions of Poland was screened to select bacteriocin-producing strains. Moreover, enzymatic activity of the enterococcal strains isolated from different healthy dogs was evaluated. In horses, we were focused mainly on the bacteria from the genera Enterococcus and Staphylococcus; among enterococci, 3 taxonomical species were identified using Maldi-Tof analysis: Enterococcus mundtii, E. faecium and E. hirae. Among staphylococcal isolates, 10 taxonomical species were identified: Staphylococcus vitulinus, S. epidermidis, S. capitis, S. pasteuri, S. xylosus, S. cohnii subsp. urealyticus, S. cohnii subsp. cohnii, S. sciuri, S. warneri a S. haemolyticus. These species belong to 4 staphylococcal groups regarding the 16S rRNA analysis. In enterococcal strains from faeces of dogs the following enzymatic activity was tested: amylolytic, pektinolytic and inulolytic. The highest measured values were in pektinolytic activity (Dr. Lauková, Dr. Strompfová, Dr. Kubašová et al.).
In the framework of the collaboration of our Institute (Laboratory of Animal Microbiology) with the Faculty of Veterinary Medicine (Technic University in Lisbon-Portugal, genes for bacteriocins-plantaricin A, plantaricin B, plantaricin C, plantaricin D, plantaricin N, plantaricin K, plantaricin R and plantaricin L were detected in Lactobacillus plantarum LP13/Ba strain isolated from faeces of pheasant. Optimalization of plantaricin A purification is processed (Dr. Lauková, Dr. Kandričáková).
Based on the collaboration of our Institute (Laboratory of Animal Microbiology) with Istituto Zooprofilattico Sperimentale delle Venezie, Viale dell Universita, Legnaro, Italy (Risk Analysis and Public Health Department) we were focused on testing of sensitivity to antimicrobial proteinaceous substances-enterocins of Campylobacter strains isolated from different poltry in Italy. Enterocins tested were those characterized in our laboratory produced by our Enterococus faecium strains. Campylobacters isolated from the poultry in Italy (39 strains; 24 strains identified as Campylobacter jejuni and 15 strains belonged to the species C. coli) were screened for their sensitivity to antibiotics as well as to enterocins. Campylobacters were sensitive to streptomycin and chloramphenicol and they were resistant to at least 2 antibiotics. The important result is information that the majority of Campylobacters (except 4 nonhemolytic strains C. jejuni) including strains resistant to antibiotics were sensitive to 8 enterocins used with inhibition activity 100 AU/ml (from 39 tested strain it was 30). Results with scientific impact were obtained; not only regarding the spreading range of inhibition spectrum of our enterocins but also from the aspect of enterocins possible utilization in prevention or in elimination of Campylobacters; there are only limited information in this area associated with inhibition activity against Campylobacters up to now (Dr. Lauková, Dr. Ščerbová).
Dr. Oceľová within internship (SAIA-CEEPUS) continued with analyses of thymol in poultry gastrointestinal tract using solid phase micro-extraction and gass chromatography/mass spectrometry in the Institute of Animal Nutrition and Functional Plant Compounds, University of Veterinary Medicine, Vienna, Austria (April-June 2016). Moreover, she has presented results gained within her previous stay in this Institute at two international conferences.
METHAGENE network unites researchers from across Europe to work on new solutions in breeding for low-emitting ruminants. The network bringing together European experts working on strategies of quantify and mitigate methane emissions from ruminants. METHAGENE supports training of early stage researchers across Europe on short-term scientific missions (STSMs). From October 12 to 14, 2016, 49 researchers from across Europe came together during the METHAGENE workshop to connect their work related to measuring and analyzing large scale individual methane emissions. The meeting took place in Padua (Italy) and three researchers from our Institute participated (Váradyová, Kišidayová, Pisarčíková). Dr. Pisarčíková presented her results within the METHAGENE-STMS in Switzerland (ETH Zürich, Switzerland) with the title „Usefulness of rumen fluid collected from slaughtered cattle to estimate within- and between-breed variation in methane emission potential in vitro“. With the financial support of METHAGENE-STMS the next young researcher Dr. Livia Kolesar Fecskeová was able to spend a month at INRA in France in laboratory of Diego Morgavi. Her host was Dr. Milka Popova and Dr. Livia Kolesar Fecskeová was participated on experiment with the title „Transcriptomic study of the metabolism of rumen methanogen“ as a part of study of ruminal methanogens (16/05/2016 – 17/06/2016, INRA, Clermont Ferrand, France). (Dr. Váradyová).
Within the study stay (SAIA, 28, September 2015 – 31, January 2016) in the laboratory of prof. Angela Calderon (Drug Discovery and Development Department, Auburn University Harrison School of Pharmacy, USA), PhD. student of Institute – Dr. J. Pisarčíková, studied the stability, metabolism and bioavailability of bioactive natural products by LC-MS.
In collaboration with Wageningen University, Wageningen, Netherlands, two new species of rumen organic acid producing actinobacteria – Actinomyces succiniciruminis a A. glycerinitolerans were characterised. Both species ferment various sugars to mainly succinate and lactate and small amounts of acetate (http://dx.doi.org/10.1016/j.syapm.2016.08.001, Dr. Pristaš). In ongoing experiments the draft genome sequence of A. glycerinitolerans G10 was obtained and accepted for publication in Genome announcements Journal.
In collaboration with The National Veterinary Research Institute, Pulawy, Poland, we characterised animal isolates of gram-negative bacteria from exotic turtles and snakes imported to Poland. The isolates were identified using molecular methods (MALDI-TOF MS, 16S rRNA analysis) and their resistance to antibiotics and plasmid content was determined. The preliminary data indicate that exotic animals could be an important source of antibiotic resistant pathogenic bacteria.