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

Short communication : Immunohistochemical study of sodium-dependent glucose co-transporters in ostriches kidneys

Saabunud / Received 02.10.2020 ; Aktsepteeritud / Accepted 20.11.2020 ; Avaldatud veebis / Published online 21.11.2020 ; Vastutav autor / Corresponding author: Piret Hussar e-mail: [email protected] of the two families of glucose transporters identified, the sodium-dependent glucose co-transporters contribute to renal glucose reabsorption. Due to the lack of knowledges of the localization of SGLTs in bird's kidneys, the present study aimed to immunolocalize Na+-glucose co-transporters SGLT1 and SGLT2 in ostrich's kidneys. In the study kidney material derived from five 14 days old female ostriches. Material 0.5–1.0 cm in diameter was fixed in 10% formalin, dehydrated, embedded into paraffin; thereafter slices 7 μm in thickness were cut and deparaffi-nized, followed by immunohistochemical staining with polyclonal primary antibodies Rabbit anti-SGLT1 and Rabbit anti-SGLT2 (Abcam, UK) according to the manufacturers' guidelines (IHC kit, Abcam, UK). Our study revealed the immunohistochemical localization of SGLT1 and SGLT2 in the proximal tubules of the renal cortex. The immunohisto-chemical locations of sodium-dependent glucose transporters resembled those in mammals

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

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

Morphometrical evaluation of some anatomical features in pig kidneys: Are they different from human kidneys?

In this study we present morphometrical evaluation of some kidney’s anatomical characteristics in two different pig breeds reared in Macedonian farms in order to determinate whether the investigated pig kidneys could be suitable as an experimental model in human medicine.The material was consisted of 156 pig kidneys taken from two adult pig breeds (96 kidneys of Landrace/Yorkshire crossbreed and 60 kidneys from breed Dalland), slaughtered at age of 5.5 months and weighting of 95 kg of mean. The kidneys were evaluated morphmetrically, considering weight, length, cranial and caudal pole width and cranial and caudal pole thickness. The data was statistically evaluated and compared with previous published findings for human kidneys.The results for the pig kidney morphometric measurements for Landrace/Yorkshire crossbreed were the following: weight 158.95g, length 12.73cm, cranial pole width and thickness: 6.39cm and 3.00cm respectively, caudal pole width and thickness 5.75cm and 2.83cm, respectively. In breed Dalland, the renal morphometric measurements showed: weight 182.98g, length 12.61cm, cranial pole width: 6.59cm and cranial pole thickness: 3.15cm, caudal pole width: 5.91cm and caudal pole thickness: 3.04cm. There were positive correlations between several variables considering the renal morphometry. Except for the length, the results of other morphmetrical measurements for Landrace/Yorkshire crossbreedwere smaller than those obtained for breed Dalland.From anatomical view despite the differences among these two breeds founded in this study, there are many similarities between the pig and human kidney morphometry. Considering that the best anatomical region for kidney implantation in humans depends on kidney’s anatomical features (length of blood vessels and urinary loop as well as the number of blood vessels) and renal morphological characteristics (weight, length, width and thickness) we can conclude that the pig kidneys from both investigated breeds in selected category could be suitable as an experimental models for endourological research

Laparoscopy-promising tool for improvement of reproductive efficiency of small ruminants

Assisted reproductive technologies are used to accelerate genetic gain and improve reproductive performances in farm animals, including small ruminants. This technologies include estrous synchronization, artifi cial insemination (AI) using fresh, frozen or sexed semen, embryo transfer (ET) using in vivo or in vitro produced embryos, and more advanced - cloning and production of transgenic animals. Diagnostic procedures, such as ultrasonography and laparoscopy, have been used as additional tools for monitoring the ovarian response to superovulatory treatment in donor animals as well as for AI and collection and transfer of embryos. The use of laparoscopy for assisted reproduction techniques in Macedonia commenced in the early 90’s, with the acquisition of a set of ,,Karl Storz” equipment. After the adoption of the required routine, our group has completed several scientifi c projects where laparoscopy was used for intrauterine inseminations as well as for recovery and transfer of embryos in both sheep andgoats. In the following period our group endeavored into introduction of laparoscopic insemination in the routine farm practice. Ovine intrauterine/intracornual insemination by frozen-thawed semen resulted with pregnancy rates of 45% and 60%, when AI was performed out of season and during the breeding season, respectively. In goats, this percentage occasionally peaked at 85%. The aim of this article is to review the status of implementation of laparoscopy in Assisted Reproduction Technologies (ART) of small ruminants and to present our experience in this field

Micronuclei and nuclear abnormalities in erythrocytes from barbel Barbus peloponnesius revealing genotoxic pollution of the river Bregalnica

The strong negative anthropogenic pressure on the River Bregalnica resulted in the increase of pollution in the river ecosystem. The aim of this study was to evaluate if aquatic pollutants generate genotoxic effects in barbel (Barbus peloponnesius). Thirty individuals were collected from three localities along the river: one control site and two locations in the middle and lower course of the river with the presence of different pollution sources. Blood smears were stained with May Grünwald/Giemsa method and the frequency of micronuclei, vacuolated nuclei, binuclei, blebbed nuclei and notched nuclei expressed in per mill was recorded. In fish collected from the control site, the investigated erythrocyte nuclear abnormalities revealed lowest values compared to polluted sites. In general, significantly higher values for erythrocyte deformations, including micronuclei, were noted in the middle and lower part of the river. Significantly higher values of vacuolated and blebbed nuclei were detected in fish from the middle course of the river, while at the lower course the frequencies of occurrence of micronuclei and notched nuclei were significant compared with control site. The middle course of the River Bregalnica has more diverse pollutants compared to the lower course, and consequently the highest frequency of all nuclear deformations was found in barbel from the middle course of the river. Obtained results suggest that pollutants draining into the Bregalnica river ecosystem display a genotoxic effect on the local barbel population

Comparative study of glucose transporters GLUT-2 and GLUT-5 in ostriches gastrointestinal tract

The knowledge about transport of sugars in animals and birds gastrointestinal tract is very important for science as carbohydrates are the main energy source of food. Since until now there is few information about the localization of glucose transporters - integral membrane proteins that mediate the transport of glucose and related substances across the cellular membranes - in birds gastrointestinal tract, the aim of the present study was to localize glucose transporters-2 and -5 (GLUT-2 and -5) in three parts of the ostriches gastrointestinal tract – proventriculus, duodenum and ileum - comparatively in ostrich chicken in their early ontogenesis period. Material from the superficial gland zone of the proventriculus, duodenum and terminal zone of the ileum were collected from eight female ostriches (Struthio camelus var. Domesticus): two chickens after hatching, three 7 and three 30-days old ostriches. The material was fixed with 10% formalin, embedded into paraffin, slices 7 μm thick were cut followed by immunohistochemical staining with polyclonal primary antibodies Rabbit anti-GLUT-2 and Rabbit anti-GLUT-5, carried out according to the manufacturers guidelines (IHC kit, Abcam, UK). The results showed that the staining for both antibodies was weaker in all parts of the gastrointestinal tract of ostriches after hatching compared to 7 and 30 days old ostriches showing that the gastrointestinal tract of ostriches immediately after hatching is not entirely capable of transportation of carbohydrates. The results of our study may indicate the possibility of a close relationship between feeding and the ability to transport sugars in the gastrointestinal tract

Pelvi-caliceal system on pig kidney: a model for urologyc procedures in human medicine

The aim of the study was to establish a valid experimental model of pig kidney that can be used in experimental works in human medicine. The pig kidney collecting system was morphologically investigated on total 72 three dimensional silicones S10 corrosion endocast. Morphometrical evaluation was performed on caudal pole collecting system anatomy considering the caudal infidibular length, caudal infidibular width and caudal infidibulopelvic angle. The morphology on pelvi-caliceal system showed two different morphological forms witch in 52.77% of cases were bilateral symmetric. The caudal infidibular length was 3.09sm and the infidibular width was measured on 0.66mm. The extent of caudal infidibulopelvic angle measured between infidibular and pelvi-uretral axis was 50.14 o degrees. According the results in this study, there are many similarities in collecting system morphology between pig and human kidney. The resemblances in collecting system anatomy conduce to conclusion that the pig kidney is good animal model that could be used for urological procedures in the field of experimental human medicine