ABSTRACT
Background
Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable ...test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality.
Results
Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics.
Conclusions
Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.
The effects of stress on processes in the body are becoming an increasingly relevant research subject. The reproductive ability of bovine animals largely depends on these effects, whilst embryo ...transfer is increasingly being used as a reproduction method. In this study, we established the differences in the implantation ability of heifers that were treated (N=17) with selenium (Se) and vitamins AD
E, and non-treated heifers. Upon transfer, we took blood samples from both groups and used the total antioxidant status (TAS) value to analyze the presence of reactive oxygen species (ROS), the levels of non-esterified fatty acids (NEFA) and the levels of vitamin E and Se in blood plasma. In the study, we were able to demonstrate that preventive measures in the form of supplementation of vitamin E and Se, mitigate the effects of oxidative stress, strengthen the ability of an organism to improve the dynamic relationship between free radicals and antioxidants, improve the energy status of cattle, positively impact reproductive parameters and increase the success rate of embryo transfer. The difference in the number of successful embryo implantations between the control and treated group was statistically significant, with 64.7% of treated heifers being pregnant after embryo transfer and giving birth to healthy calves. In the control group, the implantation success rate was 41.2%. The supplementation of antioxidants in the form of a combination of vitamin AD
E and Se, proved to be a good method for strengthening the defense of an organism and an effective mean of preventive clinical approach for improving fertility parameters.
Exposure to polychlorinated biphenyls (PCBs), which are persistent lipophilic environmental pollutants, has a variety of adverse effects on wildlife and human health, including bone mineralization, ...growth and mechanical strength. The present study evaluated the effects of lactational exposure to nondioxin-like PCB-155 and dioxin-like PCB-169, individually and in combination, on pubertal rat femur development and its biomechanics. After offspring delivery, Wistar rat mothers were divided into four groups, i.e., PCB-169, PCB-155, PCB-155+169 and control, and were administered PCBs intraperitoneally. Data on bone geometry, biomechanics and mineral composition were obtained by analysis of femurs from 42-day-old offspring by microCT scanning, three-point bending test and inductively coupled plasma mass spectrometry. Decreased somatic mass and femur size, i.e., mass, periosteal circumference and cross sectional area, were observed in the PCB-169 and PCB-155 groups. Additionally, lactational exposure to planar PCB-169 resulted in harder and more brittle bones containing higher amounts of minerals. Combined exposure to structurally and functionally different PCBs demonstrated only mild alterations in bone width and mineralization. To conclude, our results demonstrated that alterations, observed on postnatal day 42, were primarily induced by PCB-169, while toxicity from both of the individual congeners may have been reduced in the combined group.
•Effect of lactational exposure on pubertal rat femur was mainly induced by PCB-169.•Exposure to PCB-169 or PCB-155 reduced somatic and femur growth on PND 42.•PCB-169 induced stiffer and more brittle bones with higher amounts of minerals.•Combined PCB exposure induced milder alteration in studied bone parameters.
The effects of Beauvericin (BEA) produced by the fungusBeauveria bassianaandFusariumsp. on neuromuscular transmission and contractility were determined in an isolated neuromuscular mouse ...hemidiaphragm preparation. BEA (5 µM) significantly inhibits indirectly elicited twitch amplitude. At higher concentrations (7.5 and 10 µM), BEA produces a significant reduction of directly elicited, or complete block of indirectly evoked, muscle contraction. BEA also appears to be myotoxic, as indicated by a slowly developing muscle contracture. Development of neuromuscular blockade and contracture is concentration dependent. BEA acted by presynaptically depressing spontaneous acetylcholine release as indicated by the reduction in the frequency of spontaneous miniature endplate potentials (MEPPs), while the membrane potential of muscle fibers remained unchanged. At higher concentrations (7.5 and 10 µM), BEA progressively reduces or completely blocks MEPPs and EPPs amplitudes. Changes in MEPPs and EPPs are associated with substantial depolarization of muscle fibers when exposed to 7.5 and 10 µM of BEA. These results indicate that BEA has neurotoxic and myotoxic effects, which overlap in a narrow range of concentrations.
The aim of the study was to investigate the effect of a therapeutic renal diet on selected clinical, biochemical, and urinary parameters and on selected parameters of oxidative stress in cats with ...early stages of chronic kidney disease (CKD). A prospective study of a 3-month duration was conducted to evaluate the effect of renal diet on selected clinical and laboratory parameters in client-owned cats with early stages of CKD. Of a total of 29 enrolled client-owned cats, nineteen (19) cats completed the study, ten receiving renal diet and nine receiving a diet of the owner’s choice. A clinical examination was performed, and blood and urine samples were collected on the day of presentation and at regular check-ups after 3-4, 7-8, and 10-12 weeks. Serum creatinine and symmetric dimethylarginine (SDMA) concentrations and selected parameters of oxidative stress (plasma glutathione peroxidase (GPX) activity and plasma malondialdehyde (MDA) and serum selenium concentrations), were measured and electrophoresis of urinary proteins was performed. At inclusion, a significant positive correlation (p < 0.001) was found between serum selenium concentration and plasma GPX activity (Pearson correlation coefficient 0.83 (95% CI: 0.65 - 0.92) and a significant negative correlation (p < 0.001) between serum SDMA and urine specific gravity (Pearson correlation coefficient -0.70 (95% CI: -0.87 - (-0.38)). At the end of the 3-month feeding trial no significant difference was found in SDMA and creatinine concentrations.
UČINEK MONOTERAPEVTSKE LEDVIČNE DIETE NA KLINIČNE, BIOKEMIJSKE, URINSKE IN SERUMSKE PARAMETRE OKSIDATIVNEGA STRESA PRI MAČKAH S CDK STOPNJE 1 IN 2
Izvleček: Namen študije je bil raziskati učinek terapevtske ledvične diete na izbrane klinične, biokemijske in urinske parametre ter parametre oksidativnega stresa pri mačkah v začetnih stopnjah kronične ledvične bolezni (KLB). Raziskava je bila zasnovana kot prospektivna, tri mesece trajajoča klinična študija, v katero je bilo vključenih 29 lastniških mačk. Devetnajst mačk je zaključilo študijo, od teh jih je deset prejemalo ledvično dieto, devet pa vzdrževalno dieto po izboru lastnika. Pri vseh mačkah smo izvedli klinični pregled in odvzem krvnih ter urinskih vzorcev na dan vključitve v študijo in pri treh kontrolnih pregledih, ki so bili izvedeni 3–4, 7–8 in 10–12 tednov kasneje. Določili smo serumsko koncentracijo kreatinina, simetričnega dimetilarginina (SDMA) in izbrane parametre oksidativnega stresa (aktivnost plazemske glutation peroksidaze (GPX) in plazemska koncentracija malondialdehida (MDA) ter serumska koncentracija selena). Poleg tega smo izvedli elektroforezo urinskih proteinov. Ob vključitvi mačk v raziskavo smo ugotovili značilno pozitivno korelacijo (p < 0,001) med serumsko koncentracijo selena in aktivnostjo plazemske GPX (Pearsonov korelacijski koeficient 0,83, 95 % CI: 0,65–0,92) ter značilno negativno korealcijo (p < 0,001) med koncentracijo SDMA in specifično težo urina (Pearsonov korelacijski koeficient –0,70 (95 % CI: –0,87–(–0,38)). Trimesečno hranjenje s terapevtsko hrano ni privedlo do značilnih sprememb v serumski koncentraciji SDMA in kreatinina pri vključenih mačkah.
Ključne besede: klinični parametri; simetrični dimetilarginin; oksidativni stres; ledvična dieta; mačke; kronična ledvična bolezen; elektroforeza urinskih proteinov
In recent years, the less-studied
mycotoxins have attracted increasing interest due to the lack of survey data and their ability to cause toxic effects in animals and humans. To fill the gap, the aim ...of this three-year survey was to investigate the presence and co-occurrence of
and other mycotoxins in a total of 433 cereal grain samples from Slovenian farms and agricultural cooperatives from 2014 to 2016. Using the multi-mycotoxin method, 14 mycotoxins were determined. In 53% of 433 analysed samples, contamination with at least one mycotoxin was found. Deoxynivalenol (DON) and tenuazonic acid (TeA) were present in 32% and 26% of cereal grain samples, respectively, whereas alternariol (AOH), tentoxin (TEN), alternariol monomethyl ether (AME), 3- and 15-acetyldeoxynivalenol (3- and 15-AcDON), and zearalenone (ZEN) were present in fewer than 15% of the samples. Ochratoxin A (OTA) was found in one rye sample, while diacetoxyscirpenol (DAS), HT-2 and T-2 toxin, and fumonisins B
and B
(FB1 and FB2) were not detected. The highest maximum and median concentrations of
toxins were determined in spelt in 2016 (TeA, 2277 µg/kg and 203 µg/kg, respectively), and those of
toxins in wheat in 2015 (DON, 4082 µg/kg and 387 µg/kg, respectively). The co-occurrence of two or more mycotoxins was found in 43% of the positive samples. The correlations between
toxins were very weak but statistically significant (r: 0.15-0.17,
: 0.0042-0.0165). A well-known correlation between
toxins DON and ZEN was weak and highly significant (r = 0.28,
< 0.0001).
Before insects can be used widely as an alternative source of dietary protein, their allerginicity should be investigated. Therefore, the aim of our study was to assess the potential adverse ...reactions of the immune system of dogs against Tenebrio molitor proteins. Dogs sensitised to storage mites T. putrescentiae and A. siro were included. Clinically healthy and clinically allergic dogs were compared. Proteins were extracted from mealworm larvae and their digestibility determined by in vitro incubation with digestive proteases. Mealworm protein extracts and digests were analysed by SDS–PAGE. Canine sera tested for the presence of mite-specific IgEs were used for subsequent Western blotting. LC-MS/MS analysis was used to identify mealworm proteins and their allergenic potential was predicted with the AllermatchTM tool. The binding of canine sera IgEs to mealworm proteins was confirmed; however, the differences between the two groups of dogs were not significant. Moreover, no clear correlation was found between sensitisation to storage mites and clinical status of the dogs. Altogether, 17 different proteins were identified, including tropomyosin, α-amylase, and Tm-E1a cuticular protein that are known cross-reacting IgE-binding allergens. Our results suggest that dogs allergic to mites may clinically express also the cross-reactivity with mealworm proteins.
This four-year study reports the occurrence of ergot alkaloids (EAs) in cereals intended for animal feeding collected in Slovenia. A total of 517 samples of cereals were analysed using liquid ...chromatography-tandem mass spectrometry for the presence of EAs. The sample set included wheat, rye, triticale, oat, spelt and barley. The study revealed that 17% of the analysed cereal samples were contaminated with at least one ergot alkaloid. EAs have two epimeric forms: -ine and -inine. The incidence rates of the -ine and -inine forms in the analysed samples were 16% and 15%, respectively. The highest contamination rates were observed in rye (54%), oat (50%) and spelt (30%), where the highest mean concentrations of total EAs were also determined (502 µg/kg, 594 µg/kg and 715 µg/kg, respectively). However, the highest concentrations of total EAs were found in wheat and rye (4217 µg/kg and 4114 µg/kg, respectively). The predominant EAs were ergometrine, ergosine and ergocristinine. The occurrence of six or more ergot alkaloids was observed in 49% of the positive samples. A weak correlation (
= 0.284) in the positive samples was found between the mass of sclerotia and the total concentrations of EAs using the Spearman correlation coefficient.
ABSTRACTThis paper addresses yield loss due to red deer (Cervus elaphus) grazing on permanent grasslands on three organic farms in SE Slovenia over two consecutive years. Deer graze on permanent ...grasslands throughout the year, but the resulting crop loss varies during the growing season. Accounting for three mowings, the quantitative yield loss due to deer grazing totalled 48–52%, or 3.9–4.3 t/ha. Deer had the highest consumption capacity in the spring (first mowing: yield loss of 1.7–1.9 t of air-dried hay/ha), and towards the end of the growing season, consumption decreased (third mowing: loss of 0.9 t/ha). A floristic inventory indicated low species diversity and production in permanent grasslands in SE Slovenia. The crude protein content in unprotected plots was generally greater than that in protected plots, but due to a large quantitative yield loss, the yield of crude protein and net energy for lactation on heavily grazed land were much lower than those on protected land in all mowings. The average additional cost per unit of heavily grazed area ranged from 182 to 344 EUR/ha, and the differences are the result of the different intensities of game grazing in individual locations and grassland production capacities.
In a controlled field experiment (54±1 days), 20 primiparous sows (day 89±2 of gestation) were randomly divided into two equal groups. The sows were fed diets naturally containing 0.3 mg/kg of the ...mycotoxin deoxynivalenol (DON) for the control group and 5.1 mg/kgDON, 0.1 mg/kg zearalenone and 21.6 mg/kg fusaric acid for the experimental group. The sows from the experimental group consumed significantly less feed during gestation (P=0.002), during lactation (P=0.027) and in the weaning to oestrus interval (P<0.001) than control sows. Blood samples were taken four times during the experiment (day 0, 17, 42 and 52). There were no differences in total and differential blood leukocyte count, with the exception of neutrophils on day 52, which reached 42.22±9.02% in the experimental group and 32.10±10.65% in the control group (P=0.040). Flow cytometric analysis of peripheral blood T lymphocytes with monoclonal antibodies against CD3, CD4 and CD8 revealed the percent of both CD3+CD4+ and CD3+CD8+ cells. We calculated the absolute number of CD3+CD4+ andCD3+CD8+ cells (109/L), which showed a decreasing trend in the experimental group, with 2.40±0.63 and 3.83±1.15, respectively, at the beginning and 1.78±0.39 and 2.74±0.89, respectively, at the end of experiment. In the control group, these values were 2.19±0.70 and 3.41±0.87 and finally 2.12±0.69 and 3.11±1.12 x 109/L, respectively. The obtained results suggest that feed naturally contaminated with Fusarium toxins reduces the feed intake, influences the neutrophil count, and has immunomodulatory effect on T lymphocyte numbers.Key words: leukocytes; T lymphocytes; mycotoxins; deoxynivalenol; flow cytometry; sowUČINEK KRME, NARAVNO KONTAMINIRANE S TOKSINI PLESNI Fusarium sp., NA IZBRANE HEMATOLOŠKE PARAMETRE IN LIMFOCITNE PODVRSTE PRVIČ BREJIH SVINJPovzetek: V nadzorovanem poskusu, ki je na veliki farmi prašičev trajal 54±1 dan, smo 20 mladic (breje 89±2 dni) naključno razdelili v dve enaki skupini. Živali so zauživale krmo, ki je bila v kontrolni skupini naravno kontaminirana z mikotoksinom deoksinivalenolom v koncentraciji 0,3 mg/kg krme. Krma za poskusno skupino je vsebovala 5,1 mg/kg deoksinivalenola, 0,1 mg/kg zearalenona in 21,6 mg/kg fuzarne kisline. Mladice iz poskusne skupine so v obdobju brejosti (P = 0,002), laktacije (P = 0,027) in v času od odstavitve pujskov do ponovne osemenitve (P < 0.001) zaužile statistično značilno manjšo količino krme kot živali iz kontrolne skupine. Med poskusom smo mladicam štirikrat odvzeli kri (dan 0, 17, 42 in 52). V skupnem številu levkocitov in diferencialni krvni sliki med skupinama ni bilo statistično značilnih razlik, razen v številu nevtrofilcev 52. dan poskusa, ko je bila vrednost v poskusni skupini 42.22±9.02 % in v kontrolni skupini 32.10±10.65 % (P = 0.040). S pretočno citometrijo smo določili delež limfocitov T in njihovih podvrst, celic T pomagalk (CD3+ CD4+ ) in citotoksičnih celic T (CD3+ CD8+ ), ki smo jih označili z monoklonskimi protitelesi. Iz dobljenih podatkov smo izračunali absolutno število CD3+ CD4+ in CD3+ CD8+ (109 /L), ki se je v poskusni skupini zmanjševalo od 2.40±0.63 in 3.83±1.15 na začetku poskusa do 1.78±0.39 in 2.74±0.89 na koncu poskusa. Vrednosti v kontrolni skupini so bile na začetku poskusa 2.19±0.70 in 3.41±0.87 ter na koncu 2.12±0.69 in 3.11±1.12 x 109 /L. Rezultati kažejo, da zauživanje krme, naravno kontaminirane s toksini plesni vrste Fusarium, vpliva na ješčnost živali, število nevtrofilcev in ima imunomodulatorni učinek na število limfocitov T.Ključne besede: levkociti; limfociti T; mikotoksini; deoksinivalenol; pretočna citometrija; prašiči