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.
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.
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.
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.
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.
The application of immune serum is one of the most efficient method used formerly in the protection of raised piglets’/weaners’ health . The objective of the study was to determine specific antibody response during hyperimmunization of fatteners with a self-prepared subunit vaccine, and to propose production method of immune serum against Gram-negative bacteria antigens. The vaccine was administered every two weeks, 4 times. Individual and pooled serum samples were assayed for IgM, IgG and IgA antibodies against Histophilus somni recombinant Hsp60, H.somni rOMP40 and Pasteurella multocida LPS. Additionally total serum IgG and haptoglobin concentrations were measured. Two weeks after the first vaccination IgM antibody raised significantly against H.s. rOMP40 and LPS, whereas after 4 weeks it increased against rHsp60 antigens. Anti-LPS IgM antibody raised up stepwise till the end of the observation, but IgM antibody against H.s. rHsp60 and H.s. rOMP40 decreased in further samplings. A significant raise in IgG class H.s. rHsp60- -antibody was found 4 weeks after the first immunization and a similar raise against two remain- ing antigens after 6 weeks. The intensity of the reaction increased till the end of the experiment. The raise in IgA antibody level was observed only for H.s. rHsp60 antigen. Clinically observed, proper animal health and welfare were confirmed by haptoglobin concentration, which remained in physiological range. At least 4 booster doses were necessary to obtain hyperimmune serum containing a high level of antibodies against examined antigens. The number of immunizations influenced response profiles for specific IgM, IgG, IgA antibodies.
The lithostratigraphy, biostratigraphy, sequence stratigraphy, ammonite and inoceramid faunas of the Upper Albian, Cenomanian, and Lower Turonian Karai Formation, the highest unit of the Uttatur Group in the Pondicherry Sub-Basin of the Cauvery Basin in Tamil Nadu, south India, are documented. Detailed logs and descriptions of sections between Karai and Kulakkalnattam, Odiyam and Kunnam, and north-west of Garudamangalam are presented. They provide the evidence for an ammonite zonal scheme that can be correlated in detail with sequences developed in Europe, with successive Upper Albian zones of Pervinquieria (Subschloenbachia) rostrata and P. (S.) perinflata (the latter on slight evidence), Cenomanian zones of Mantelliceras mantelli, Cunningtoniceras cunningtoni, Calycoceras (Newboldiceras) asiaticum, Pseudo calycoceras harpax, Euomphaloceras septemseriatum and Pseudspidoceras footeanum. The Lower Turonian is represented by a Neoptychites cephalotus–Mytiloides borkari fauna. Over 120 ammonite species are described, of which Puzosia (Bhimaites) falx, Protacanthoceras parva, Watinoceras elegans, Euomphaloceras varicostatum, Kamerunoceras multinodosum, and Carthaginites multituberculatus are new. The new genus Kunnamiceras, with Ammonites tropicus Kossmat, 1865 as type species, is interpreted as a paedomorphic dwarf derivative of Pseudocalycoceras harpax (Stoliczka, 1864). Ammonite faunas from shales are dominated by feebly-ornamented taxa: leiostraca; those from sandstones by strongly ornamented taxa: trachyostraca, differences interpreted as reflecting the preferred habits of adults in life. 15 species of inoceramid bivalves, including a newly described species Inoceramus chiplonkari, are recognised, with a mixed East African–Euramerican–North Pacific affinity. On the basis of the stratigraphic framework developed, a sequence stratigraphic interpretation of the Karai Formation is proposed, and correlated with those recognised in Europe, Morocco, and the United States Gulf Coast and Western Interior.
In the current study, twenty lambs, aged 4 months, half male and half female, were classified into four groups, with five in each group. The experimental three groups of lambs were given intravenous (IV), intramuscular (IM) and subcutaneous (SC) administrations of recombinant ovine interferon-τ (roIFN-τ). The fourth group (normal control) of lambs was given normal saline injections in the same way. After administrations, blood samples were collected from the tested animals at different time points post injection, and the serum titers of roIFN-τ were measured using cytopathic effect (CPE) inhibition bioassay. The results of calculating pharmacokinetic (PK) parameters using DAS software showed that the PK characteristics of roIFN-τ through IV injection conformed to the two-compartment open model, whose half-life of distribution phases (T1/2α) was 0.33±0.034 h and the elimination half-life(T1/2β) was 5.01±0.24 h. However, the PK features of IM injection and SC injection of roIFN-τ conformed to the one compartment open model, whose Tmax were 3.11±0.26 h and 4.83±0.43 h, respectively, together with an elimination half life(T1/2β) of 9.11±0.76 h and 7. 43±0.58 h, and an absorption half-life (T1/2k(a)) of 1.13±0.31 h and 1.85±0.40 h, respectively. The bioavailability of roIFN-τ after IM administration reaches 73.57%, which is greater than that of SC administration (53.43%). These results indicate that the drug administration effect can be preferably obtained following a single dose IM administration of the roIFN-τ aqueous preparation. This study will facilitate the clinical application of roIFN-τ as a potential antiviral agent in future work.
Animals as a source of organs and tissues for xenotransplantation could become a backup solution for the growing shortage of human donors. The presence of human xenoreactive anti- bodies directed against Galα1,3Gal antigens on the cell surface of a pig donor triggers the activa- tion of the complement leading to a hyperacute reaction. The development of genetic engineer- ing techniques has enabled the modification of genomes by knocking in and/or knocking out genes. In this paper, we report the generation of modified pigs with ZFN mediated disruption of the GGTA1 gene encoding the enzyme responsible for synthesis of Galα1,3Gal antigens. ZFN plasmids designed to target the exon 9 region of the pig GGTA1 gene encoding the catalytic domain were injected into the pronuclei of fertilized egg cells. Among 107 piglets of the F0 gene- ration analyzed, one female with 9-nt deletion in exon 9 of the GGTA1 gene was found. 13 of 33 piglets of the F1 generation represented the +/- GGTA1 genotype and 2 of 13 F2 piglets repre- sented the -/- GGTA1 genotype. No changes in the animals’ behavior, phenotype or karyotype were observed. Analysis confirmed heredity of the trait in all animals. A complex functional analysis of the modified animals, including flow cytometry, human serum cytotoxicity test and immunohistochemical detection, was performed to estimate the phenotype effect of genetic modification and this indicated an efficient GGTA1 knock-out in modified pigs.
Introduction: Platelet-rich plasma (PRP) preparations can be used in bone tissue healing but there are numerous doubts among clinical orthopedists about effectiveness of this method. Materials and methods: The studies were carried out in 12 rabbits of white termond breed. In operating room we operationally generated cylindrical, unicortical defects of the diameter of 4 mm in the middle of the shafts of both femurs. The defects in the left bones were left without filling and served as controls, and 0.7 ml of the ready-to-use PRP was administered to the defects in the right bones (experimental group). We evaluated the usefulness of the diagnostic methods applied: biomechanical tests, micro-CT tests, densitometry, typical radiology, macroscopic measurements, histopathological examinations. Results: The macroscopic measurements showed a statistically significant increase in the dimension in the area of the right defect filled with PRP in relation to the control group. In experimented group, the assessment of the X-ray images showed the formation of a callus cuff around the defects. Densitometric examinations showed no statistically significant differences between defects in the experimental and control group. The analysis of the micro-CT examina- tions showed an increase in the total volume of the tissue examined (Vb) and the low density tissue fraction (Vb2) in the experimental group. The biomechanical examinations revealed signi- ficant decrease in the maximum breaking force (F max) necessary to break the bone in the experi- mental group in relation to the control group. Conclusions: Platelet-rich plasma (PRP) stimulates bone formation in the area of bone defects and may accelerate bone regeneration.
Sperm-mediated gene transfer (SMGT) is based on the ability of spermatozoa to bind exoge- nous DNA and transfer it into oocytes by fertilization. However, SMGT is still undergoing opti- mization to improve its efficiency to produce transgenic animals. The acrosome reaction is neces- sary for spermatozoa to carry the exogenous DNA into oocytes. In this study, the effect of the acrosome reaction on the efficiency of spermatozoa carrying exogenous DNA was evalua- ted. The results showed that the efficiency of the acrosome reaction was significantly higher (p<0.05) after incubation with 50 μmol/L progesterone compared to incubation without proges- terone. It was significantly higher (p<0.05) in the 20, 40, and 60 min of progesterone treatment groups than in the 0 min treatment group. The spermatozoa were further incubated with cyanine dye Cy5 labeled DNA (Cy5-DNA) for 30 min at 37°C, and positive fluorescence signals were detected after the acrosome reaction was induced by progesterone at concentrations of 0 and 50 μmol/L for 40 min. The percentage of positive Cy5-DNA signals in spermatozoa was 96.61±2.06% and 97.51±2.03% following exposure to 0 and 50 μmol/L progesterone, respective- ly. The percentage of partial spermatozoa heads observed following combination with Cy5-DNA was 39.73±3.03% and 56.88±3.12% following exposure to 0 and 50 μmol/L progesterone, respec- tively. The ratio of positively stained spermatozoa combined with exogenous DNA showed no reduction after the acrosome reaction. These results suggest that the acrosome reaction might not be the key factor affecting the efficiency of SMGT.
Mechanical ventilation (MV) is a supportive and life-saving therapy, however, it can cause ventilator-induced lung injury as a common complication. Thus, recruitment manoeuvres (RM) are applied to open the collapsed alveoli to ensure sufficient alveolar surface area for gas exchange. In the light of the fact that positive pressure ventilation is currently the standard treat- ment for improving pulmonary function, extrathoracic negative pressure is considered as an alter- native form of respiratory support. The aim of this study was to estimate the proinflammatory and oxidative response during MV and lung injury as well as the response after RM. All studied parameters were assessed at the following time points: T1-spontaneous breathing, T2- MV, T3- lung injury, T4 –RM. During MV (T2) elastase, MPO, ALP release, nitrite and superoxide generation significantly increased, whereas in later measurements a decrease in these values was noted. The MDA plasma concentration significantly (p<0.05) increased at T2, reaching a level of 13.30±0.87 nmol/ml; at other time points the values obtained were similar to the baseline value of 9.94±0.94 nmol/ml, whereas a gradual decrease in SOD activity at time T2-T4 points in comparison with the baseline value was found. During the study both neutrophil activity and oxi- dative stress indicate exacerbated response after MV and lung injury by bronchoalveolar lavage; however, extrathoracic negative pressure system as the MR ameliorates damaging changes which could further lead to serious lung injury.
Concentrations of four trace elements, copper (Cu), zinc (Zn), manganese (Mn) and seleni- um (Se), have thus far proven to be affected by lentiviral infections in people and rhesus monkeys. As small ruminant lentivirus (SRLV) infection is responsible for one of the most important goat diseases, caprine arthritis-encephalitis (CAE), we evaluated serum and liver concentrations of Cu, Zn, Mn, Se in goats severely affected by symptomatic CAE and compared them with litera- ture reference intervals. Serum and liver samples of dairy goats euthanized due to severe clinical form of CAE were collected and screened for the concentration of Cu, Zn, Mn (54 serum sam- ples, 22 liver samples), and Se (36 serum samples, 22 liver samples) using flame atomic absorption spectrometry for Cu, Zn, Mn and graphite furnace atomic absorption spectroscopy for Se. In both serum and liver samples concentration of Zn was the highest, followed by Cu concentration, and then by Mn and Se. There was no relationship between serum and liver concentrations of trace elements. Liver concentrations of all four trace elements and serum Cu concentration fell within literature reference intervals, although liver Se concentration was mainly in the lower marginal range (between 0.4 and 1.0 mg/L). Serum Zn concentration was elevated (>1.2 mg/L) in all goats, serum Mn concentration was elevated (>0.04 mg/L) in 42 (78%) goats and serum Se concentra- tion was elevated (>1.6 mg/L) in 13 (36%) goats. Concluding, severe symptomatic CAE does not appear to be associated with the level of any of the four trace elements.