Determination of cytogenetic markers for biological monitoring in coypu (Myocastor coypu)

Cytogenetic tests are used to assess the influence of physical and chemical factors with potential mutagenic and genotoxic properties on the animal organism. The test results make it possible to eliminate mutagens, as well as helping predict possible genetic consequences in animal cells and assess animal resistance. The aim of this study was to examine, using cytogenetic tests, the spontaneous chromosome and DNA damage in coypu lymphocytes. Four tests: fragile site (FS), bleomycin (BLM), micronucleus, (MN) and comet were used for the first time in coypu cells. The averages with standard deviations obtained in the research were as follows: 3.30 ± 0.80 fragile sites/cell; 0.63 ± 0.80 BLM damage/cell; 6.10 ± 0.53% binucleated cells with MN; and 3.24 ± 0.63% DNA in tail. The present analysis showed high interindividual variation in spontaneous chromosomal and DNA damage levels. In the case of micronucleus, fragile sites, and comet assays, the differences between animals were statistically significant. The data suggest that these assays are sensitive enough to detect some effects on an individual animal and can be proposed as tools for coypu biomonitoring.     

The immunohistochemical evaluation of selected markers in the left atrium of dogs with end-stage dilated cardiomyopathy and myxomatous mitral valve disease – a preliminary study

Background

Dilated cardiomyopathy (DCM) and myxomatous mitral valve disease (MMVD) are the most common diseases noted in dogs. Although their pathogenesis varies, both include a significant enlargement of the left atrium.The study was carried out on left atrial specimens obtained from 56 dogs, including those from 34 dogs with clinically diagnosed MMVD, 15 dogs with DCM and 7 dogs without heart disease (control group). Dogs in the MMVD and the DCM groups presented with left atrial enlargement and stage D heart failure. The specimens underwent immunohistochemical examination using desmin, vimentin, periostin and caspase-3 antibodies.

Results

There were alterations in the expression of the studied proteins in the study groups compared to the control group. The changes included: irregularity of desmin cross-striation and desmosomes, a higher amount of vimentin-positive cells, a change in the periostin expression pattern from cytoplasmic to extracellular, and a lower expression of caspase-3. The alterations were more pronounced in the DCM group than in the MMVD group.

Conclusions

During heart failure, the pattern of desmin, vimentin, periostin and caspase-3 expression alters in the left atrium, regardless of the cause. The changes are more pronounced in dogs with DCM than in dogs with MMVD and similar left atrial enlargement, suggesting that volume overload may not be the only cause of myocardial changes in DCM.

     

Histological and morphometric evaluation of the urethra and penis in male New Zealand White rabbits

This stereudy aimed at performing a histological and morphometric evaluation of the urethra and penis of male rabbits. Seven male New Zealand White rabbits weighing 2.1–3 kg were used in the study. The whole urethra, from the urinary bladder to the external urethral orifice, was dissected from the rabbits, and the tissue was sliced into sections at an interval of 2 mm. The sections were stained with haematoxylin–eosin, Masson–Goldner trichrome stain, Van Gieson's stain and Movat–Russell modified pentachrome stain. A detailed evaluation of the morphology and morphometry was performed. The parameters assessed were the type and height of epithelium, thickness and composition of connective tissue, and thickness and structure of muscularis. The histological structure of the rabbit urethra was found to be similar to humans. However, although the rabbits were found to have the same type of penis as the humans, the internal structure of the corpora cavernosa, the relative thickness of the tunica albuginea and the rudimentary glands of the penis were found to differ in these animals. The results of the present study may be useful in the designing of implants, drug testing or surgical procedures on the physiological and pathological urethra.     

Giblets and abdominal fat of pomegranate seed oil fed chickens as a source of bioactive fatty acids

The aim of present study was to determine whether chickens’ (broilers Ross 308, n = 180, sex ratio 1:1) diet modification with different doses of grape or pomegranate seed oil will favourable change fatty acids and cholesterol content in selected giblets (liver and heart) or wastes (adipose tissue). It was also verify whether generated changes would make the giblets and wastes more valuable as dietary components or by-products for food industry. From 22 to 42 day of life, five diets were administered to chickens. Control grower diet (CON) contained 5% of soy oil, whereas in the experimental grower diets part of soy oil (1.5% or 2%) was replaced with specific amount of grape or pomegranate seed oil (GRAP 1.5; GRAP 2.0; POM 1.5; POM 2.0 respectively). Fatty acids and cholesterol content were determined with gas chromatography with flame-ionization detection. Pomegranate seed oil improved fatty acids profile more favourably than grape seed oil, which makes it a valuable additive in chickens’ feeding. Abdominal fat of pomegranate seed oil supplemented chickens appeared to be the richest sources of rumenic acid and n3 polyunsaturated fatty acids, which allows to suggest its use in manufacturing of meat products to obtain foodstuffs rich in those essential nutrients. In principal component analysis (PCA), two principal components: PC1 and PC2, which were enough to explain 29.91% of variance of initial variables, allowed to a good separation of chickens fed with both doses of pomegranate seed oil from animals from control and grape seed oil fed groups. Because poultry addresses all nutritional, institutional and consumer requirements, enrichment of giblets in rumenic acid by pomegranate seed oil incorporation into chickens’ diet may provide a valuable dietary source of bioactive fatty acids for consumers, especially of low-income countries.     

Assessment of selenium bioavailability from high-selenium spirulina subfractions in selenium-deficient rats

It was previously found that the bioavailability of Se from Se-rich spirulina (SeSp) was lower than that from selenite or selenomethionine when fed to Se-deficient rats. The present study examined the bioavailability of Se from SeSp subfractions: a pellet (P) issuing from the centrifugation of a suspension of broken SeSp and a retentate (R) resulting from ultrafiltration of the supernatant through a 30 kDa exclusion membrane. Animals were fed a torula yeast based diet with no Se (deficients) or supplemented with 75 microg of Se/kg of diet as sodium selenite (controls) for 42 days. Se-deficient rats were then repleted for 56 days with Se (75 microg/kg of diet) supplied as sodium selenite, SeSp, P, or R. During this period, controls continued to receive sodium selenite. Speciation of Se in subfractions showed that the majority was present in the form of high molecular weight compounds; free selenomethionine was only a minor constituent. Gross absorption of Se from sodium selenite, P, and R was not different and was higher than from SeSp. Only retentate allowed full replenishment of Se concentration in liver and kidney (as did sodium selenite) and glutathione peroxidase (GSHPx) activity in liver, kidney, plasma, and erythrocytes. The bioavailabilities of Se in retentate, as assessed by slope ratio analysis using selenite as a reference Se, were 89 and 112% in the tissue Se content and 106-133% in the GSHPx activities. SeSp and P exhibited a gross bioavailability of <100%. These results indicate that Se in retentate is highly bioavailable and represents an interesting source of Se for food supplementation.     

Element Concentrations in Soils and Plant Bioindicators in Selected Habitats of the Holy Cross Mountains,Poland

The objectives of this investigation in the HolyCross Mountain region of Poland were (1) to determineconcentration ranges and (2) mobility of elements within fivedetailed soil profiles and their apparent impact on thechemistry of some plant bioindicators, i.e. lichen thalli ofthe species Hypogymnia physodes (L.) Nyl. and Scotspine (Pinus sylvestris L.) bark. The results ofelemental determinations performed on soils and vegetationfrom the Holy Cross Mts show that some concentrations ofelements are elevated primarily due to air pollution.However, since 1994 a steady decrease in the content of S andsome heavy metals has been recorded in all the mediaexamined. Concentrations of many elements in H.physodes thalli and P. sylvestris needles of the Holy Cross Mountains are generally similarto those in other areas of Europe.     

Quantitative analysis, occurrence, and stability of (E)-1-(2,3,6-Trimethylphenyl)buta-1,3-diene in wine

A stable isotope dilution assay has been developed for quantification of (E)-1-(2,3,6-trimethylphenyl)buta-1,3-diene (4) in wine using a [(2)H(6)]-analogue. Using this method, 4 was found in 96 out of 97 white wines, but in none of 12 red wines analyzed. 4 was found to be most prevalent in Semillon wines, followed by Chardonnay, with Riesling showing the least amount of 4 among these three varieties. 4, like 1,1,6-trimethyldihydronaphthalene (TDN, 3), appears to be formed during the aging process. 4 was found to be unstable in model wine, and in both white and red wine, with the order of stability being model > white > red. In a PVPP-stripped red wine, the rate of degradation of 4 was substantially lessened, with the final concentrations very close to those observed in model wine. When treated with either grape or wine tannin extracts in model wine, the concentration of 4 was found to decrease to levels very close to those observed with an untreated red wine. When white wine was heated at 45 degrees C, 4 was formed, indicating the presence of precursor forms. The amounts formed were much higher than those found in a commercial white wine. 4 was also observed in red wine heated to 45 degrees C, but the concentration produced was much less than that with white wine.     

Effect of Boiling in Water of Barley and Buckwheat Groats on the Antioxidant Properties and Dietary Fiber Composition

In recent years, there has been an ever-increasing interest in the research of polyphenols obtained from dietary sources, and their antioxidative properties. The purpose of this study was to determine the effect of boiling buckwheat and barley groats on the antioxidant properties and dietary fiber composition. Antioxidative properties were investigated using methyl linoleate model system, by assessing the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity and metal chelating activity. The results were compared with butylated hydroxytoluene (BHT). Raw barley and buckwheat groats extracts showed higher DPPH scavenging ability compared to boiled barley and buckwheat groats extracts. Raw barley groats extract exhibited higher antioxidant activity than boiled groats extract in the methyl linoleate emulsion. Higher chelating ability in relation to Fe (II) ions was observed for boiled groats extracts as compared to raw groats extracts. BHT showed small antiradical activity and metal chelating activity, while showing higher antioxidative activity in emulsion system. The analysis of groats extracts using HPLC method showed the presence of rutin, catechin, quercetin, gallic, p-hydroxybenzoic, p-coumaric, o-coumaric, vanillic, sinapic, and ferulic acids. Differences in the content of dietary fiber and its fractions were observed in the examined products. The highest total dietary fiber content was detected in boiled buckwheat groats, while the lowest - in boiled barley groats. The scientific achievements of this research could help consumers to choose those cereal products available on the market, such as barley and buckwheat groats, which are a rich source of antioxidative compounds and dietary fiber.     

Exposure to non‐ionizing electromagnetic radiation of public risk prevention instruments threatens the quality of spermatozoids

The use of artificial insemination in cattle breeding has evolved to global extent, and insemination doses are often shipped via air transport which requires strict radiation‐based examinations. For the determination of effect of non‐ionizing radiation (NIR), to which are beings frequently exposed due to protection of airport or cultural event security, freshly ejaculated and cryopreserved bovine spermatozoa were used as experimental model. Following radiation with hand‐held metal detector in various exposition times (0, 10 s, 15, 30 and 60 min—groups FR, FR10, FR15, FR30 and FR60) the spermatozoa underwent motility and DNA fragmentation analyses. Study on cryoconserved semen treated with NIR was performed in time intervals 0, 10 s, 1 and 5 min (insemination doses radiated before cryoconservation—CB, CB10, CB1, CB5; samples radiated after freezing—CA, CA10, CA1 and CA5). Fresh semen and insemination doses radiated after cryoconservation showed significantly lower total and progressive motility. No effect on motility parameters was detected in semen extended with cryopreservative medium and radiated prior to freezing. Surprisingly, NIR showed a potential to stimulate spermatozoa velocity; however, the effect was modulated throughout the post‐thawing incubation. Based on the DNA fragmentation assay, sperm DNA stayed intact. Present study underlines the potential harm of NIR, which is frequently used in everyday life, with overall adverse impact on human and animal reproduction. Current study also points out on interesting short‐term spermatozoa stimulation induced by NIR.