The aim of the study was to develop new laparoscopic technique for repeated recovery of sheep oocytes. Oocytes were aspirated with specifically designed catheter. It allowed to recover oocytes without ovary damage and to preserve very good quality of recovered oocytes. Fifteen ewes were oocytes donors. Oocytes were collected: one time (group I, n=15), two times (group II, n=15), three times (group III, n=10), four times (group IV, n=5). The endoscope was inserted into the abdominal cavity. Two trockars for putting the manipulators were inserted 15 cm cranial from the udder. Oocytes were collected by aspiration of the follicular fluid from the ovarian follicles. The observed clinical complications were: ovary bleeding and cicatrix at place of needle insertion, the fragmentary adhesion of infundibulum and ovary, adhesions of omentum and peri- toneum near the place where the grasping forceps were inserted and adhesion of ovary and uterus. Ovarian follicles (n=204) were aspirated, 130 (63.8%) oocytes were obtained. Out of 130 obtained oocytes, 112 were qualified for in vitro maturation. The remaining 18 oocytes (13.8%) were rejected due to cytoplasmic changes. The proposed technique allows for the collecting oocytes of good quality that can be used for IMV/IVF techniques and cloning.
Polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite, dichlorodiphenyldichloroethylene (DDE) can disturb the secretory function of the ovary and both contractions and secretory function of the uterus during the estrus cycle and pregnancy. Additionally, PCBs can pass through the placental barrier into allantoic and amniotic fluid. The presence of PCBs in these fluids is associated with higher frequency of spontaneous abortions and premature births in humans and animals. Therefore, the effect of PCBs, DDT and DDE on the connexins (Cx26, Cx32 and Cx43) and keratin 8 (KRT8) expression in bovine placentomes was investigated. The placentome slices from the second trimester of pregnancy were incubated with PCB153, 126, 77, DDT and DDE (each at doses of 1, 10 or 100 ng/ml) for 48 h. Then, the slices were stained using immunohistochemistry. The density of Cxs staining was measured with Axio- Vision Rel. 4.8 software in fetal-maternal connections and binuclear cells (BNC). None of the tested xenobiotics (XBs) affected the localization of Cxs and KRT8 in the fetal-maternal connection area, but the XBs affected the density of Cxs in fetal-maternal connections and binuclear cells (BNCs). Depend on the doses, in fetal-maternal connections all used PCBs changed the protein expression of different Cxs, while in BNCs, all tested XBs except DDT increased the expression of Cxs. None of investigated XBs affected on KRT8 expression. In summary, used XBs affect the expression of Cxs and change the quantitative relationships between them. Therefore, XBs can unfavorably influence function of the utero-placental barrier in cows.
Although the Antarctic has avoided the worst effects of alien species, its future seems endangered due to increasing natural and man-made pressures. Rapid changes in three major environmental variables have occurred in the Antarctic region during the last decades. In the short term terrestrial biota are likely to benefit from reduced environmental stresses, but in the long run the colonization of the region by lower latitude species with greater competitive ability will become increasingly important and can lead to large-scale changes in biological composition and trophic complexity in some existing Antarctic terrestrial ecosystems. Moreover, the recent dynamic climate changes combined with human activities in the Antarctic region might modify the status of several alien species which have hitherto been considered transient or persistent and could, therefore, become naturalized and threaten the native communities on a larger scale than today, or influence the status of naturalized species.
There were tested microorganisms in differents soils at Admiralty Bay region. The physiological groups of microorganisms were restricted by the kind of organic matter. There were found in ornithogenic soils in higher number the following groups of microorganisms; proteolytic bacteria, uric acid and L-asparagine ammonifying bacteria, chitin degrading bacteria, lecithin degrading bacteria and calcium phosphate dissolving bacteria. The nitrifying bacteria were found in lower horizons of ornithogenic soils in higher number. The nitrogen fixing bacteria were found in mineral soils covered by plant associations, only. The spore-forming bacteria were detected in ornithogenic soils and in soil influenced by man.
Matrix metalloproteinases 2 and 9 (MMP2 and MMP9) are proteolytic enzymes involved with extracellular matrix degradation. They play a role in tumor invasion and metastases. Be- cause of their ability to degrade signaling molecules presented in extracellular matrix, MMPs contribute to tumor proliferation and apoptosis. The aim of this study was to evaluate expression of MMP2 (latent and both active and latent forms) and MMP9 (active, latent, active and latent forms) in different subtypes of canine lymphomas and their relationship with proliferative (mi- totic index and percentage of Ki67-positive cells) and apoptotic (apoptotic index) markers. Ex- pression of MMPs was assessed immunohistochemically using an immunoreactive score system. Expression of both MMPs was found in all 20 examined lymphomas belonging to six subtypes. Most cases showed a moderate level of all analyzed forms of MMP2 and MMP9. High expres- sion of MMPs was found in single cases. Except for a positive correlation between the active form of MMP9 and the mitotic index for all lymphoma cases, no other correlations between any remaining forms of MMPs and neither proliferative nor apoptotic markers were found, irrespec- tive of whether the analysis encompassed all cases or the most numerous lymphoma subtypes i.e. centroblastic and Burkitt-like. Our results were not able to clearly confirm the influence of MMPs on the proliferation and apoptotic activity of canine lymphoma cells. However, further studies examining MMPs activity by zymography, expression of their inhibitors and other factors in- volved in activation of cell proliferation and apoptosis inhibition are needed to clarify the role of MMPs, especially the active form of MMP9, in the behavior of canine lymphoma cells.
The high pressure die casting technology allows the production of complex casts with good mechanical properties, with high production repeatability within narrow tolerance limits. However, the casts are somewhat porous, which may reduce their mechanical properties. There are several recommendations for reducing the porosity of casts, which are aimed at setting the technological parameters of the casting cycle. One of the primary and important ways to reduce the porosity and air entrapment in the melt is a suitable gating system design. Submitted contribution is devoted to assessing the influence of the runner branching geometry on the air entrapment within the cast volume during the filling phase of the casting cycle. Four variants of the gating system for a particular cast are compared with different design of main runner branching. The initial design is based on a real gating system where the secondary runner is connected to the main runner at an angle of 90 °. The modified designs are provided with a continuous transition of the main runner into the secondary ones, with the change in the branching runner radius r1 = 15 mm, r2 = 25 mm and r3 = 35 mm. The air entrapment in the melt is assessed within the cast volume behind the cores, which have been evaluated as a critical points with respect to further mechanical treatment. When designing the structural modification of geometry it was assumed that by branch changing using the radius value r3 = 35 mm, the melt flows fluently, and thus the value of the entrapped air in the volume of the cast will be the lowest. This assumption was disproved. The lowest values of entrapped air in the melt were found in the casts with runner transition designed with radius r1 = 15 mm. The conclusion of the contribution explains the causes of this phenomenon and from a designing point of view it presents proposal for measures to reduce the entrapment of the air in casts.
When a certain Buddhist monk was once setting out on a long, difficult journey, he chose as his traveling companion a servant who was known for having a rebellious and quarrelsome nature. When asked why, he replied that he wanted to practice patience and humility. One might say that the three years when I headed the international CHIHE project were a similar lesson in virtues.
Aim: The aim of this study was to analyze the effect of bovine follicular fluid on the survival, morphology and kinetic parameters of bovine thawed spermatozoa under laboratory conditions. Materials and methods: The semen from 5 bulls of proven fertility was incubated in follicular and physiological fluid for 8 hours. During this time assessment using the CASA system was performed. At the beginning and the end of incubation process evaluation by flow cytometry was conducted. Results: The results of the sperm motility assessment showed a significant decrease in the analyzed parameters both in the follicular and physiological fluid. A significant reduction in all parameters characterizing movement properties in the semen incubated in the follicular fluid was found. In the physiological fluid, a similar trend was demonstrated only for the following proper- ties: VAP, VSL, VCL, ALH, BCF. A significant difference was found for both fluids in: VCL (p=0.026), ALH (p=0.038) and LIN (p<0.001) at the beginning of incubation. The results of the plasma membrane integrity assessment showed a statistically significant increase in the percent- age of dying sperm at the 8th hour of the incubation in the follicular fluid. In the case of semen incubation in physiological fluid, a statistically significant decrease in the percentage of live non-damaged cells was found with a simultaneous increase in the subpopulation of undamaged dead cells. Conclusions: Follicular fluid rapidly accelerates the capacitation process. The results of flow cytometry support the hypothesis concerning the ability of follicular fluid to prolong sperm sur- vival.
Despite many phytochemical and pharmacological investigations, to date, there are no reports concerning the antibabesial activity of extracts of A. millefolium against B. canis. This study was aimed at investigating the biological activities of A. millefolium against the Babesia canis parasite and to identify its chemical ingredients. The water (WE), ethanol (EE) and hexane/acetone (H/AE) extracts of plant aerial parts were screened for total phenolic content (TPC), total flavo- noid compound (TFC), DPPH free radical-scavenging activity and its antibabesial activity assay. In this study, imidocarb diproprionate was used as a positive control. The H/AE and EE extracts were analysed using gas chromatography–mass spectroscopy (GC–MS). In the EE extract, the main compounds were 17.64% methyl octadec-9-ynoate, 16.68% stigmast-5-en-3-ol(3α,24S) and 15.17% hexadecanoic acid. In the H/AE extract, the main com- pounds were 34.55% 11-decyldocosane, 14.31% N-tetratetracontane, 8.22% β-caryophyllene, and 7.69% N-nonacosane. Extract of EE contained the highest content of phenolics followed by H/AE and WE. The concentration of flavonoids in EE, H/AE and WE extracts showed that TFC was higher in the EE samples followed by H/AE and WE. The antioxidant activities were highest for AA, followed by EE, WE and H/AE. The antibabesial assay showed that the WE, EE and H/AE extracts of A. millefolium were antagonistic to B. canis. At a 2 mg/mL concentration, it showed 58.7% (± 4.7%), 62.3% (± 5.5%) and 49.3% (± 5.1%) inhibitory rate in an antibabesial assay, respectively. Considering these results, the present findings suggest that A. millefolium extracts may be a potential therapeutic agent and that additional studies including in vivo experiments are essential.
The Silurian Pelplin Formation is a part of a thick, mud-prone distal fill of the Caledonian foredeep, which stretches along the western margin of the East European Craton. The Pelplin Formation consists of organic carbon- rich mudstones that have recently been the target of intensive investigations, as they represent a potential source of shale gas. The Pelplin mudstones host numerous calcite concretions containing authigenic pyrite and barite. Mineralogical and petrographic examination (XRD, optical microscopy, cathodoluminoscopy, SEM-EDS) and stable isotope analyses (δ13Corg, δ13C and δ18O of carbonates, δ34S and δ18O of barite) were carried out in order to understand the diagenetic conditions that led to precipitation of this carbonate-sulfide-sulfate paragenesis and to see if the concretions can enhance the understanding of sedimentary settings in the Baltic and Lublin basins during the Silurian. Barite formed during early diagenesis before and during the concretionary growth due to a deceleration of sedimentation during increased primary productivity. The main stages of concretionary growth took place in yet uncompacted sediments shortly after their deposition in the sulfate reduction zone. This precompactional cementation led to preferential preservation of original sedimentary structures, faunal assemblages and early- diagenetic barite, which have been mostly lost in the surrounding mudstones during burial. These components allowed for the reconstruction of important paleoenvironmental conditions in the Baltic and Lublin basins, such as depth, proximity to the detrital orogenic source and marine primary productivity. Investigation of the concretions also enabled estimation of the magnitude of mechanical compaction of the mudstones and calculation of original sedimentation rates. Moreover, it showed that biogenic methane was produced at an early-diagenetic stage, whereas thermogenic hydrocarbons migrated through the Pelplin Formation during deep burial.
Placoid and polyodontode scales of stem chondrichthyans have been found in the early Lochkovian “Ditton Group” of the Brown Clee Hill district, Shropshire, England and at Talgarth, south Wales. One of the forms is assigned to a new species of Altholepis Karatajūtė-Talimaa, 1997, a genus already recognised from Lochkovian shallow marine deposits in Celtiberia, Spain and the Northwest Territories, Canada as well as the type locality in Podolia, Ukraine. Altholepis salopensis sp. nov. is based on small polyodontode scales with typically three to eight high odontodes; the scale form was previously considered to belong to acanthodian “Nostolepis” robusta (Brotzen, 1934). The structure of other scales formerly assigned to “Nostolepis” robusta has led us to erect a new genus Jolepis for this scale form, which differs from Altholepis in lacking an ordered layout of odontodes. Jolepis robusta (Brotzen, 1934), originally (and possibly still) considered to be an acanthodian, is also known from the Baltic countries, Russia, and northern Germany (ex erratic limestones). Scales of acanthodian Parexus recurvus Agassiz, 1845, and/or possibly from the stem chondrichthyan Seretolepis elegans Karatajūtė-Talimaa, 1968 (scales of these two taxa are barely distinguishable), and of stem chondrichthyan Polymerolepis whitei Karatajūtė-Talimaa, 1968 are also present. Altholepis, Jolepis gen. nov., Seretolepis Karatajūtė-Talimaa, 1968 and Polymerolepis Karatajūtė- Talimaa, 1968 are found in marine deposits elsewhere; the British occurrence of these taxa adds to the debate on the sedimentological origins of the Lower Old Red Sandstone deposits in the Welsh Borderland. The geographic range of several early sharks is now known to extend around the Old Red Sandstone continent and beyond.
The Lower Devonian ‘Placoderm Sandstone’ in the Holy Cross Mountains (HCM) is filled with abundant impressions of disarticulated vertebrate remains. The only acanthodian macroremains named to date are fin spines of Machaeracanthus polonicus Gürich. Fin spine impressions in slabs from the Winna Formation (Emsian) at Podłazie Hill (near Daleszyce) in the southern HCM, and also the Barcza Formation (?Lochkovian) at Barcza Quarry, Miedziana Góra Conglomerate (?Lochkovian), Gruchawka, and Zagórze Formation (middle–upper Emsian) at Bukowa Mountain in the northern HCM, reposited in the University of Warsaw, Polish Geological Institute-National Research Institute, Warsaw, and Natural History Museum, London collections, have been cast and studied in order to better document this poorly known taxon. As noted in other Machaeracanthus species, we have found that M. polonicus has two different morphotypes of spines, which abut lengthwise to form a pair of spines. Our investigations show that the fin spine assemblage includes Onchus overathensis as well as M. polonicus, and probably another undetermined acanthodian. The affinities of O. overathensis are reassessed. It is here considered to be a diplacanthiform, and reassigned to the genus Striacanthus, as S. overathensis. Acanthodian scapulocoracoids have also been identified, as well as tightly spiralled toothwhorls which could be from an acanthodian.
During thirty three expeditions to the Polish Arctowski Antarctic Station signifi− cant influences of human activity upon the environment have been recorded. Introductions of alien species, shifts of bird and seal breeding areas and decreases in both bird and seal populations, are the most obvious effects of human pressure. Though numbers of visits by tourists have increased during this period, impacts from expeditioners appear to be the main cause of changes. In particular, increasing numbers and mobility of summer groups at the station are the likely most influential factors.
Net of temperature, salinity and dissolved oxygen content measurements were made in situ at 24 stations in Admiralty Bay with the use of automatic analyzer Martek MK IV. Results proved a presence of two water masses located horizontally one over the other, with the boundary at depth of 15—35 m. The main, lower mass consists of homogenous waters inflowing from the Bransfield Strait. The upper, thin and much differentiated layer is formed through interaction of the main underlying water mass with meltwaters from glaciers and sea ice. It is probably formed along the whole archipelago and carried by surface currents into the bay where it is subjected to further modifications. Local salinity and temperature extremes are associated with glacier water runoff; local dissolved oxygen maxima seem to be connected with phytoplankton distribution. Strong currents occurring in te bay due to water circulation may cause local, short term and sometimes considerable fluctuations in values of parameters recorded at individual stations.
Blastocystis is a common enteric protozoan of humans and various species of animals. Culture and microscopic examination of fecal samples is the conventional method for identifying four major forms of Blastocystis (vacuolar, granular, non-vacuolar or cystic). In this article, we compared eight liquid media for cultivation of Blastocystis spp. Study material included fecal samples from clinically healthy pigs. Significant differences in the growth of Blastocystis on individual media were observed.
Faecal Enterococcus hirae from domestic ducks were studied for their bioactivity to select bioactive strain for more detailed study with its probable use in poultry and also to bring novelty in basic research. After defecation, faeces (n=23, faecal mixture of 40 ducks) were sampled from domestic ducks in eastern Slovakia; birds were aged from eight to 14 weeks. E. hirae strains were identified using Matrix-assisted laser desorption/ionization time-of flight mass spectrometry with a highly probable species identification score (2.300-3.000) or a secure genus identification/ /probable species identification score (2.000-2.299), confirmed by polymerase chain reaction and phenotypization in accordance with the properties for the type strain E. hirae ATCC 9790. Strains were hemolysis negative (γ-hemolysis), and did not have active enzyme stimulating disorders. Enterocin genes were detected in three strains out of seven. Three out of four Enterocin genes were detected in Kč1/b (Ent A, P, L50A); the most frequently detected was the Ent P gene. The strains inhibited indicator strains E. faecalis, listeriae, but also Escherichia coli and Buttiauxiella strains. Lactic-acid producing E. hirae were mostly susceptible to antibiotics. Based on parameter evaluation, E. hirae Kč1/b, Kč6 can be additionally studied to select the type of bioactive substance.
Brachiopods are reported for the first time from the Lower Miocene Cape Melville Formation of King George Island, South Shetland Islands, West Antarctica. Two genera, Liothyrella Thomson and Paraldingia Richardson have been identified. This is the first occurrence of Paraldingia in Antarctica.
A hybrid artificial boundary condition (HABC) that combines the volume-based acoustic damping layer (ADL) and the local face-based characteristic boundary condition (CBC) is presented to enhance the absorption of acoustic waves near the computational boundaries. This method is applied to the prediction of aerodynamic noise from a circular cylinder immersed in uniform compressible viscous flow. Different ADLs are designed to assess their effectiveness whereby the effect of the mesh-stretch direction on wave absorption in the ADL is analysed. Large eddy simulation (LES) and FW-H acoustic analogy method are implemented to predict the far-field noise, and the sensitivities of each approach to the HABC are compared. In the LES computed propagation field of the fluctuation pressure and the frequency-domain results, the spurious reflections at edges are found to be significantly eliminated by the HABC through the effective dissipation of incident waves along the wave-front direction in the ADL. Thereby, the LES results are found to be in a good agreement with the acoustic pressure predicted using FW-H method, which is observed to be just affected slightly by reflected waves.
Function of duck (Anas platyrhynchos) major histocompatibility complex class I (Anpl-MHC I) molecules in binding peptides is through the peptide binding groove (PBG), which is thought to be influenced by the high polymorphism of α1 and α2 domains. However, little is known about the polymorphism of Anpl-MHC I peptide binding domain (PBD), especially in the domestic duck. Here, we analyzed the polymorphism of forty-eight Anpl-MHC I α1 and α2 domains from domestic duck breeds previously reported. All sequences were analyzed through multiple sequence alignment and a phylogenetic tree was constructed. The coefficient of variance of the peptide binding domains (PBDs) from WS, CV, JD, and SX duck breeds was estimated based on the Wu-Kabat variability index, followed by the location of the highly variable sites (HVSs) on reported crystal structure models. Analysis of α1 and α2 domains showed common features of classical MHC class I and high polymorphism, especially in α1 domain. The constructed phylogenetic tree showed that PBDs of domestic ducks did not segregate based on breeds and had a close phylogenetic relationship, even with wild ducks. In each breed, HVSs were mostly located in the PBG, suggesting that they might determine peptide-binding characteristics and subsequently influence peptide presentation and recognition. The combined results of sequence data and crystal structure provide novel valuable insights into the polymorphism and diversity of Anpl-MHC I PBDs that will facilitate further studies on disease resistance differences between duck breeds and the development of cytotoxic T-lymphocyte (CTL) epitope vaccines suited for preventing diseases in domestic ducks.
In this study, we propose a possible way of obtaining reproductive and respiratory syndrome virus (PRRSV) free offspring from genetically valuable lines of Přeštice black-pied boars com- ming from PRRSV-positive pig breeding herds with the use of artificial insemination (AI). The ejaculates were collected from 4 different lines of boars. Samples of fresh semen were not detected with the virus and 12 sows were inseminated. Blood samples of sows and their offspring were repeatedly tested for the virus but the results were negative. We managed in this way to maintain the endangered population of this breed and obtain PRRSV-free offspring.
Several human studies have reported that capsaicin has anti-pruritic effects. Moreover, sever- al concentrations of topical capsaicin have been used to alleviate itch. The aim of this study was to investigate the anti-pruritic effect of capsaicin against histamine-induced pruritus compared with that of topical steroid or vehicle in 15 healthy beagles. Fifteen dogs were divided into three groups (n = 5 each), and treated topically with one of the following on the left side of the neck: capsaicin, positive control (steroid), or negative control (vehicle). Each treatment was performed twice daily for 8 days. All dogs were injected with histamine intradermally before treatment and on the 2nd, 4th, 6th, and 8th days of the treatment to evoke itch. Pruritus, wheal, and erythema intensity were assessed at each evaluation; cutaneous temperature was also recorded. On the final day, skin biopsy was conducted for histopathological evaluation for all dogs. The severity of pruritus was lesser in the capsaicin-treated group compared with the negative control group on day 8 (p<0.05). In the capsaicin and steroid groups, wheal size, erythema index, and cutaneous temperature also decreased compared with pretreatment. Histopathological evaluation showed that the capsaicin-treated group had a higher number of inflammatory cells in the dermis com- pared to the vehicle control group; however, the steroid-treated group showed less severe inflam- matory reactions than the vehicle control group. These results suggest that capsaicin cannot reduce inflammation but may play a helpful role in reducing pruritus in dogs.
Dendritic cells (DCs) due to their ability to present antigens are essential during the immune response to infections. The aim of the study was to evaluate the influence of bovine leukaemia virus (BLV) infection on DC properties. Cytokine profiles of myeloid, plasmacytoid and mono- cyte derived DCs from BLV infected cattle were analysed. Concentrations of IL-6, IL-10, IL-12, IFN-γ, and TNF-α in DC cultures were measured by flow cytometry. Obtained results indicated activation of pDCs population, where a significant increase in production of the IFN-γ was shown. Meanwhile, a decrease in production of IFN-γ and increase in production of IL-10 were shown in mDCs; the main population responsible for antigens presentation. This may indicate a contribu- tory role of the population during the process of persistent infection. In MoDCs population a significant elevation in secretion of proinflammatory cytokines – IL-6 and TNF-α was noted.
Meteorological conditions at Arctowski Station during 2013–2017 were presented against the background of regional climate changes, especially air temperature decline. Air temperature, relative air humidity, air pressure, solar radiation, wind speed and direction, snow cover and precipitation were collected with an automatic weather station and manual measurements and were further analysed. The obtained results were compared with data from previous years and with data from other stations located on King George Island. Our observations confirm that the vicinity of Arctowski Station experienced a decrease in air temperature during summer, which supports the hypothesis of regional cooling.