Effect of t-2 mycotoxin on glut-2 expression in the broiler's gastrointestinal tract

T-2 toxin is a trichothecene mycotoxin. It is a naturally occurring mold by product of Fusarium spp. fungus which is toxic to humans and animals. Deleterious effects of these feed contaminants in animals are well documented, ranging from growth impairment, decreased resistance to pathogens, hepato- and nephrotoxicity to death. As the initial interaction of mycotoxins is with the gut epithelium the epithelium is repeatedly exposed to these toxins at higher concentrations than other tissues. Several methods for the determination of T-2 toxin based on traditional chromatographic, immunoassay, or mass spectroscopy techniques have been described. As up to now there is lack of knowledges about the effect of T-2 mycotoxin on the expression of glucose transporters in the proximal part of the gastrointestinal tract (GIT), the aim of our investigation was to study the experssion of glucose transporter-2 (GLUT-2) in the epithelial cells of glandular stomach and duodenal mucosa comparatively in normal and T-2 toxicated broilers. Material from the two parts of GIT was collected from six 7-day-old normal female broilers and six 7-day-old female broilers with T-2 toxicosis. The specimen were fixed with 10% formalin, embedded into paraffin, slices 7 μm thick were cut followed by immunohistochemical staining with polyclonal primary antibody Rabbit anti-GLUT-2 carried out according to the manufacturers quidelines (IHC kit, Abcam, UK). The results showed the moderate expression of GLUT-2 in epithelial cells both in the glandular as well as in the duodenal mucosa in the control group’s chicken and the weaker expression of GLUT-2 after T-2 toxin administration indicating to the reduced glucose transport in the gastrointestinal system during T-2 mycotoxicosis

T-2 mycotoxin induced apoptosis in broiler’s liver tissue

Apoptosis is a process of programmed cell death that occurs in multicellular organisms. As T-2 toxin is known to induce apoptosis in mammalian cells, the aim of the present experiment was to study the toxic effect of T-2 on chicken liver tissue using apoptosis-related antibodies p21 and p53 which are involved in the p53/p21-mediated apoptotic signalling pathway. The experiment was conducted on fourteen 40-day-old broilers (Gallus gallus domesticus) who were divided into control and T-2 toxin groups. For the T-2 toxin group, T-2 toxin (Sigma, Germany) was dissolved in water and given per os for three consecutive days. The material of the liver was taken 24 hours after the last application. The specimens were fixed with 10% formalin and embedded into paraffin; slices 5 μm in thickness were cut followed by immunohistochemical staining with polyclonal primary antibodies p21 and p53 (Abcam, UK) according to the manufacturer’s guidelines (IHC kit, Abcam, UK). Strong expression of p21 and p53 found in hepatocytes, endotheliocytes and around blood vessels together with large tissue destructions in T-2 toxin group birds’ liver indicates apoptosis and histopathological changes in liver tissue during T-2 mycotoxicosis

Morphofunctional characteristics of the adrenal cortex in tesetctomised rats acclimated on different ambient temperature

The activity of the adrenal gland is connected with the activity of other endocrine glands, such as the gonads. The experiments were conducted on testectomized adult, male rats, Wistar strain, acclimated on two ambient temperatures: room and moderately high temperature. Bilateral testectomy was performed on the rats of both temperatures and analyses were made 15-20 days after surgery. It was shown that the adrenal weight of testectomized heat acclimated rats was significantly increased. The widening of zona reticularis, shortening of zona fasciculata, as well as presence of supracortical nodules in the adrenal cortex of testectomized rats, regardless of the ambient temperature, was evidenced. Lipid content in the adrenocortical cells was lowered in both groups, but that was more prominent in testectomized rats from room temperature. The morphometric measurements of the nuclear area and volume were significantly elevated only in zona reticularis cells of testectomised animals from room temperature. Those parameters were significantly decreased in testectomized heat acclimated rats in comparison with the same values in the testectomized animals from room temperature. These results suggest that testectomy provokes changes towards increased activity of the adrenal cortex in both ambient temperatures, but those changes are less intense in heat acclimated animals

Influence of genetic selection on the myofibre type composition of porcine biceps femoris muscle: a comparative study of a purebred (Nero di Parma) and commercial hybrid pigs (Large White × Landrace × Duroc)

This study is aimed to evaluate histological features related to the different quality of meat cuts obtained from different pig breeds. We compared animals genetically selected to restore the original local purebred “Nero di Parma” pig, and highly selected fast-growing commercial hybrids. As weight is the main factor determining the time of slaughter, we compared equally weighing animals, sampled from slaughter for edible use, regardless of their age and sex, and immunohistochemically demonstrated the myofibre type composition of their biceps femoris muscle. In both groups we observed type I myofibres, situated as central islets, encircled by type IIA, IIX and IIB myofibres ordered in concentric rings according to the dynamic of their differentiation/maturation. However, the purebred pig muscles contained a smaller quantity of myofibres expressing the MyHC-IIb isoform, related to rapid postmortem glycolytic rate, meat toughness and poorer quality, in comparison to commercial hybrids. This proves that the latter are subjected to a more rapid transition in the expression of the different MyHC, probably as a consequence of genetic selection and breeding conditions, such as different feeding and housing reducing the possibility of continuous physical exercise. Further studies on the postnatal transitions timing of myosin heavy chain isoforms in functionally different muscles of various breeds are necessary to verify if they might be “artificially modulated”, with the aim to design breeding programs allowing a good balance between growth performance, muscularity and meat quality

Adverse effect of dexamethasone on development of the fetal rat ovary.

Dexamethasone (Dx) is often used in obstetric practice to promote fetal lung maturation and to prevent respiratory distress syndrome when the risk of preterm delivery persists. This therapy enables survival of the newborn, but also is associated with deleterious effects on the offspring, such as reproductive disorders. The aim of this study was to determine specifically whether prenatal exposure to Dx disturbs the physiological balance between proliferation and apoptosis of germinative cells (GC) in the ovary of 19 and 21 day old fetuses and thus induces developmental programing of the female reproductive system. Pregnant Wistar rats (n = 10/group), separated into control (vehicle) and Dx-treated (0.5 mg/kg body mass) groups, received injections on gestational days 16, 17 and 18. Exposure to Dx lowered the volume of the fetal ovary by 30% (p<0.05) in 21 day old fetuses, as well as the total number of GC in the ovary by 21% (p<0.05). When compared to the controls, in Dx-exposed fetuses the total number of PCNA positive GC was 27% lower at 19 days and 71% lower at 21 day old (p<0.05), while total numbers of caspase-3 positive GC were 2.3 fold and 34% higher respectively, (p<0.05). Our results demonstrate that prenatal exposure to Dx diminished proliferation but increased the rate of germinative cell apoptosis, with consequently reduced total germinative cell number and ovary volume. Impairment of fetal oogenesis and fewer GC in the fetal ovary compromise the oogonial stock and thus may constitute a risk for female fertility. This article is protected by copyright. All rights reserved.This is the peer reviewed version of the following article: Ristić N, Nestorović N, Manojlović-Stojanoski M, Trifunović S, Ajdžanović V, Filipović B, Pendovski L, Milošević V. Adverse effect of dexamethasone on development of the fetal rat ovary. Fundam Clin Pharmacol. 2018, which has been published in final form at [http://doi.org/10.1111/fcp.12415]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions

The Effect of Moderate Heat on Rat Pituitary ACTH Cells: Histomorphometric, Immunofluorescent and Hormonal Study

In areas with moderate continental climate, increased average ambient temperature during the summer represents a stressogenic factor that affects the hypothalamo-pituitaryadrenocortical axis in mammals. Therefore, we wanted to examine the effects of 4 days of constant exposure to moderately elevated ambient temperature (35 ± 1oC) on the histomorphometric and immunofl uorescent characteristics, as well as on the hormonal secretion of pituitary corticotropes (ACTH) cells in adult male rats. In comparison with the controls kept at 20 ± 2oC, a signifi cant increase (p<0.05) of the absolute and relative pituitary weight (23.1% and 36.1%, respectively) was registered after exposure to heat. The localization, as well as the shape of the ACTH cells in the heat exposed group was not signifi cantly altered, but their immunopositivity was weaker. After 4 days of heat exposure, a weaker signal confi rmed the relative fl uorescence intensity of the ACTH cells (15.3%, p<0.05). In heat exposed rats, an increase of the cellular and nuclear volumes of immunolabelled ACTH cells and decrease of their volume density (6.9%, 14.3% and 20.0%, respectively; p<0.05) was registered. Observed histomorphometric and immunofl uorescent features of the pituitary ACTH cells were in accordance with the increased (p<0.05) value of plasma adrenocorticotropic hormone (ACTH) by 23.7% compared to the control rats. It can be concluded that the 4-day exposure to moderately elevated ambient temperature intensifi es pituitary ACTH secretion in adult male rats

Histological and morphofunctional parameters of the hypothalamic-pituitary-adrenal system are sensitive to daidzein treatment in the adult rat.

The isoflavone, daidzein is a biologically active, plant-derived compound that interacts with estrogen receptors. Data from previous studies have suggested that daidzein exerts beneficial effects in many diseases; however, as an endocrine disrupter, it may also alter the functioning of the endocrine system. Data regarding the effect of daidzein on the morphofunctional and histological parameters of the hypothalamic-pituitary-adrenal (HPA) system is still lacking. Therefore, using the newCAST stereological software, we investigated the effects of chronic (21 days) daidzein treatment on corticotropin-releasing hormone (CRH) neurons within the hypothalamus and corticotropes (ACTH cells) in the pituitary, while image analysis was employed to-examine the intensity of fluorescence of CRH in the median eminence (ME) and adrenocorticotropin hormone in the pituitary in adult orchidectomized (Ovx) rats. Circulating ACTH and corticosterone levels were also analyzed. This study showed that daidzein treatment decreased the volume density of CRH neurons within the paraventricular nucleus as well as CRH immunofluorescence in the ME. The total number of ACTH cells was decreased, while ACTH cell volume and the intensity of ACTH fluorescence were increased following daidzein treatment. Both ACTH and corticosterone blood levels were increased after daidzein administration. The results of performed experiments clearly demonstrate that volume density of CRH neurons; total number and volume of ACTH cells, as well as stress hormones levels are vulnerable to the effects of daidzein

Anatomske varijacije vratnih mišića u pasa.

Muscle variations are common in humans, as well as in both wild and domestic animals. They are defined as additional muscle bundles or bellies, unusual insertions or even complete absence of the muscle. In this manuscript, we have described different variations of neck muscles in dogs. Fifty-seven medium sized dogs of different breeds, sex and ages were dissected between 2005 and 2011, within gross anatomy course. These variations were observed in four (7.02%) dogs and have included the sternohyoideus, stylohyoideus, sternocephalicus and scalenus dorsalis muscle. One dog presented three muscle variations (left sternohyoideus muscle, left and right stylohyoidues muscle), another two dogs showed two muscle variations (left and right sternocephalicus muscle) and one dog had one muscle variation (left scalenus dorsalis muscle). Muscle variations were observed bilaterally three times and once involved the stylohyoideus muscle and in the other two cases, the sternocephalicus muscle. The close phylogenetic relationship explains the observed muscle variations. Our description of neck muscle variations in dogs is a contribution to phylogenetic research and, in the terms of veterinary medicine, their presence has to be taken in account during surgical procedures of the neck region.Varijacije mišića učestale su u ljudi, kao i u divljih i domaćih životinja. Varijacije mišića mogu se definirati kao pojava dodatnoga mišićnog traka ili trbuha, neuobičajeno mjesto početka ili završetka mišića kao i potpuni nedostatak mišića. U ovom radu opisali smo različite varijacije mišića u pasa. U razdoblju između 2005. i 2011. godine na nastavi anatomije secirano je 57 pasa srednje veličine, različite pasmine, spola i dobi. Varijacije mišića vrata uočili smo u 4 (7,02%) psa na mišićima: m. sternohyoideus, m. stylohyoideus, m. sternocephalicus i m. scalenus dorsalis. Na jednom psu pronašli smo tri varijacije (lijevi m. sternohyoideus te lijevi i desni m. stylohyoidues), na druga dva psa dvije varijacije (lijevi i desni m. sternocephalicus) te na jednom psu jednu varijaciju (lijevi m. scalenus dorsalis). Bilateralne varijacije mišića uočene su triput i uključuju m. stylohyoideus u jednom i m. sternocephalicus u dva slučaja. Uočene varijacije mišića mogu se obrazložiti bliskim filogenetskim odnosom. Opis varijacija mišića u ovom radu naš je doprinos filogenetskim istraživanjima, a u okvirima veterinarske medicine, njihova se prisutnost mora uzeti u obzir tijekom kirurških zahvata u području vrata

Improved determination of macroscopic parasite preparations using S10 modified plastination procedure

Macroscopic preparations of parasites fixed in formaldehyde or alcohol don’t fulfill in complete the requests for education, as well as their determination, mainly because of the toxic fumes and not enough visible structure of fixed parasite. Using the modified С10 plastination method, parasites from three different phylum were prepared: Plathelminthes: Class Cestoda (Dipilidum caninum, Moniezia spp and larvae from T.Echinococcus granulosus - Echinococcus unilocularis, larvae from T. pisiformis - Cysticercus pisiformis, , larvae from T. hidatigena - Cysticercus tenuicollis), Phylum Nemathelminthes, Class Nematoda, (Ascaris suum, Macracanthorhynchus hirudinaceus, Diro filaria immitis), Phylum Arthropoda, Class Arachnida (tick from the Ixodidae family) and Class Insecta (Gasterophilus intestinalis, Hypoderma bovis). The aim of this study was conserving the parasites in native condition with plastination method and improved determination according to their visible morphologic structure. Parasites were previously kept in 10% formaldehyde. Prepared parasites were dry, chemical free, not toxic and safe for the environment, flexible and with detained form and structure. There was a variation in the natural colors in some of the parasites, as a result from long-time formalin fixation. Preparations made with this method are permanent educative material which enables improved study of parasite’s structure

Morphological peculiarity of the renal parenchyma on S10 thin plastinated pig kidneys

The aim of this study is to investigate the morphological structures on the renal parenchyma on the pig kidneys, prepared in thin slices by S10 sheet plastination method. A total number of 60 kidneys taken form two adult breeds are plastinated in 2mm sagital thin sections. The morphological structure on thin kidney slices is analyzed and their anatomic-topographical relationship is investigated. The prepared thin kidney slices are permanent, flexible, dry, and odorless with smooth surfaces anatomical models with clear distinction between renal medulla and renal cortex. In cross-bread landras/yorkshire, the number of renal pyramids is ranged between 8-14 (average 10.63) while in breed dalland the number is ranged between 8- 13(average 9.94)(p>0.05). Three morphological forms are found in pig kidneys based of the variation of adhesion of renal pyramids and derange of their renal papilla into renal pelvis. According the results can be concluded that the S10 sheet plastination method could be used for preparing of thin anatomical models that are suitable for education and research purposes enabling three-dimensional plan view of anatomical structures inside of kidneys