In homeostasis, which plays an important role in the proper functioning and maintenance of the internal functioning of the body, kidneys play a key role in being responsible for the proper homeostasis of glucose. Among glucose transporters, sodium-dependent glucose co-transporters (SGLTs) have a major role in the kidney's ability to reabsorb glucose. Although the localization of these transporters has been extensively studied in mammals, there are still gaps in knowledge of the localization of SGLTs in birds of different age groups. The aim of this study was to immunolocalize in kidneys of hen chickens of different ages the sodium-dependent glucose co-transporters SGLT1 and SGLT2, comparing the localization between different age groups. The kidneys derived from 32 hen chickens ( Gallus gallus domesticus) ) were devided equally into four age groups: 3, 7, 14, and 20 day old broilers, 8 indivuals in each group. The polyclonal primary antibodies Rabbit anti- SGLT1 and Rabbit anti-SGLT2 (Abcam, UK) were used together with the corresponding IHC kit (Abcam, UK). The results were visualized photographically using an AxioCam HRc camera (Germany) connected to a Zeiss Axioplan-2 Imaging microscope (Germany). The study revealed similar immunolocalization of SGLT1 and SGLT2 in the apical part of cells of proximal renal tubules in hen chickens' kidneys in all age groups. Strong staining of SGLT2 was noted also in the cytoplasm of epithelial cells of the proximal straight and convoluted tubules. Based on our study, the kidney tissue of newly hatched chickens is ready immediately after hatching for glucose reabsorption and transport, similarily to that of three-week-old chicks.

The kidney plays an essential role in the proper homeostasis of glucose. In the kidney, glucose transport is carried out across cell membranes by two families of glucose transporters—facilitated diffusion glucose transporters (GLUTs) and Na(+)-dependent glucose co-transporters (SGLT family). Among the transporters, sodium-dependent glucose co-transporters play a major role in the kidney‘s ability to reabsorb glucose. Although the localization of glucose transporters has been extensively studied in mammals, there are still knowledge gaps regarding the localization of SGLTs in birds. The aim of this research was to conduct a comparative study of the immunolocalization of the sodium-dependent glucose co-transporters SGLT1 and SGLT2 in the kidneys of healthy and T-2-mycotoxicated chickens. Immunohistochemical staining was carried out using the polyclonal primary antibodies SGLT1 and SGLT2 (Abcam, UK) in kidney tissue derived from seven healthy and seven T-2-mycotoxicated 7-day-old female layer-type Ross chickens (Gallus gallus domesticus). The sections were stained using an immunohistochemistry kit (Abcam, UK). In the kidneys of the healthy birds, strong staining of SGLT1 and SGLT2 was observed in the cytoplasm of the epithelial cells of the proximal straight and convoluted tubules. In the kidneys of the birds of the T-2 toxin group, weak expression of SGLT1 and SGLT2 with morphological changes occurred, indicating reduced glucose transport in the urinary system during T-2 mycotoxicosis.

In the changing internal and external conditions, maintenance of a constant internal environment – homeostasis – plays a significant role in the proper functioning of the organism. Kidneys play a key role in the homeostasis of glucose, in which the sodium-dependent glucose co-transporters contribute to renal glucose reabsorption. Although the localisation of Na+-glucose co-transporters has been extensively covered in animals’ kidneys, the localisation of the transporters in birds’ kidneys is still understudied. The purpose of this study was to immunolocalise the sodium-dependent co-transporters SGLT1 and SGLT2 in kidneys of ostrich chickens of different ages. In the study, kidney material derived from fifteen ostriches was divided equally into three age groups – 1-, 7-, and 14-days-old ostrich chickens. The polyclonal antibodies Rabbit antiSGLT1 and Rabbit anti-SGLT2 (Abcam, UK) served as primary antibodies and were used together with the IHC kit (Abcam, UK). With the AxioCam HRc camera (Germany) connected to the microscope Zeiss Axioplan-2 Imaging (Germany), the photos were taken and saved to the computer. As the result of the study on ostrich chickens of different ages, SGLT1 was noted to be localised in the renal straight proximal tubules and SGLT2 in the proximal convoluted tubules of nephron. The immunohistochemical locations of sodium-dependent glucose co-transporters revealed to be similar in ostriches’ kidneys of all age groups. The staining for SGLT2 was noted to be more intensive compared to the staining for SGLT1. As avian kidneys have unique morphological and functional features compared to animals, it is recommended that further studies would be performed on the renal tissue of different avian species

As traumatic brain injury (TBI) is one of the major causes of permanent disability, there is increasing interest in the long-term outcome of TBI. While motor deficits, cognitive impairment and longer-term risks of neurodegenerative disease are well-established consequences in animal models of TBI, pain is discussed less often despite its high prevalence. The current study addresses the need to characterize the extent of chronic pain and long-term behavioral impairments induced by moderate lateral fluid percussion injury (latFPI) in mice up to 12 months post-TBI and evaluates the validity of the model. Adult male BALB/c mice were subjected to latFPI, and the results were compared with outcomes in sham-operated mice. Mouse behavior was assessed at 1 and 7 days and 1, 3, 6, 9, and 12 months post-injury using sensory-motor (neurological severity score, NSS), cold (acetone) and mechanical sensitivity (von Frey), depressive-like behavior (tail suspension), locomotor (open field), motor coordination (rotarod) and cognitive (Morris water maze, y-maze, passive avoidance) tests. Animals with TBI demonstrated significantly higher NSS than the sham-operated group for up to 9 months after the injury. Cold sensitization was significantly increased in the contralateral hind paw in the TBI group compared to that of the sham group at 3, 6, and 9 months after TBI. In the von Frey test, the withdrawal threshold of the contralateral and ipsilateral hind paws was reduced at 6 months after TBI and lasted for up to 12 months post-injury. latFPI induced progressive depressive-like behavior starting at 6 months post-injury. No significant deficits were observed in memory, motor coordination or locomotion over the 12-month assessment period. The present study demonstrates that moderate TBI in mice elicits long-lasting impairment of sensory-motor function, results in progressive depression and potentiates peripheral pain. Hence, the latFPI model provides a relevant preclinical setting for the study of the link between brain injury and chronic sequelae such as depression and peripheral pain.

Studies on localization and distribution of enteroendocrine cells (EECs) are important for better understanding of their role in the ontogenetic development of intestines. Information about the distribution of the most important endocrine cells in the digestive tract of the ostrich is very limited; therefore, the aim of the present study was to identify gastrin and somatostatin EECs in the small intestine of the ostrich (Struthio camelus var. domesticus) chicks at different ages. Six embryos along with 42 ostriches of both sexes from hatching up to 60 days post-hatching, including six embryos, were obtained from an ostrich farm in Latvia. Duodenum, jejunum and ileum were investigated using routine histology and immunohistochemistry methods. Gastrin and somatostatin EECs were examined in 10 microscopic fields of the intestinal mucosa in each tissue sample. The cells were detected in all age groups as well as the embryos. The number of both types of EEC in the mucosa of the ileum was significantly lower (p < .01-.05) than in the duodenum. The present study suggested that the EEC may have a role in the development of the mucosa of the intestinal tract of ostriches with possible involvement in the development of the digestive functions.

Although patterns of glucose transporter expression and notes about diseases leading to adaptive changes in intestinal fructose transport have been well-characterized, the connection between infection and fructose transportation has been lightly investigated. Up to now only few studies on GLUT-5 expression and function under pathological conditions in bird intestines have been carried out. The aim of our current research was to immunolocalize GLUT-5 in chicken duodenal epithelium in norm and during T-2 mycotoxicosis. Material from chicken (Gallus gallus domesticus) duodenum was collected from twelve seven-day-old female broilers, divided into control group and broilers with T-2 mycotoxicosis. The material was fixed with 10% formalin and thereafter embedded into paraffin; slices 7 μm in thickness were cut, followed by immunohistochemical staining, according to the manufacturers guidelines (IHC kit, Abcam, UK) using polyclonal primary antibody Rabbit anti-GLUT-5. Our study revealed the strong expression of GLUT-5 in the apical parts of the duodenal epithelial cells in the control group chickens and weak staining for GLUT-5 in the intestinal epithelium in the T-2 mycotoxicosis group. Our results confirmed decreased the expression of GLUT-5 in the duodenal epithelium during T-2 mycotoxicosis.

Out 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 deparaffi­nized, 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 immunohisto­chemical locations of sodium-dependent glucose transporters resembled those in mammals.

High-yielding cow rations under intensive production conditions contribute to the development of subacute rumen acidosis (SARA), leading to pathologies such as rumenitis, laminitis, reproductive disorders, loss of productivity and reduced longevity. The aim of this work is to develop a reticulo-ruminal long-acting cyber-physical system for monitoring rumen parameters. Three scientific institutions in cooperation with two farmers conduct the research in order to create a prototype of a low-power wireless sensor network system for early diagnostic of subacute rumen acidosis of dairy cows. The new system architecture includes, reticulo-ruminal bolus with pH and temperature sensors, a microcontroller, a radio transmitter and a power supply module. The system includes a base station for data collection from boluses, an MQTT broker, a web server and a database. Data communication solution has been developed and tested in the laboratory, and micro-controllers have been selected and adapted for data processing. In addition, research is under way to create an autonomous long-term power supply system. Work shall be conducted in two directions: (a) stand-alone battery-powered electricity supply system; (b) an autonomous power supply system based on the generation of an electrostatic generator. The results of the initial stage of the research are discussed in this paper.

Although hexoses glucose and fructose serve as important energy sources of food, up to now, there is little information about hexose transporters in birds’ intestinal epithelium during their first postnatal week. The aim of the investigation was to carry out an immunohistochemical study of integral membrane proteins glucose transporter-2 and -5 (GLUT-2 and GLUT-5) on intestinal epithelial cells of ostrich chicks during their first postnatal week. The material from duodenum and ileum was collected from 9 female ostriches (Struthio camelus var. Domesticus) divided into three age groups, three birds in each group: chicks immediately after hatching, 3-day-old ostriches and 7-day-old chicks. The material was fixed in 10% formalin, embedded into paraffin, slices 7μm 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 manufacturer’s guidelines (IHC kit, Abcam, UK). Immunohistochemical localization of GLUT-2 and -5 in the intestinal epithelial cells in ostriches of different age groups was determined. In the groups of chicks after hatching and 3-day-old ostriches, enterocytes in duodenal epithelium were mostly unstained and goblet cells stained weakly for both antibodies. Weak staining of enterocytes and goblet cells was also noted in the ileal epithelium of the chick after hatching. Moderate staining of goblet cells was noted in the 3-day-old chicks’ ileal epithelium. In 7-day-old ostriches, the expression of both antibodies was weak in duodenal but moderate in ileal epithelial cells. The pattern of immunohistochemical expression of GLUT-2 and GLUT-5 in ostriches’ intestinal epithelial cells confirms our hypothesis that the intestinal tract of ostriches after hatching is not yet entirely capable of transportation of hexoses and showed that it is completing gradually during the first postnatal week.

Abstract The study characterises canine adipose-derived stem cells (cASCs) in comparison to human ASCs (hASCs) and tests their safety in a canine model after intravenous administration. cASCs from two dogs were cultured under hypoxic conditions in a medium supplemented with autologous serum. They were plastic adherent, spindle-shaped cells that expressed CD73, CD90, and CD44 but lacked CD45, CD14, HLA-DR, and CD34. cASCs differentiated toward adipogenic, osteogenic, and chondrogenic lineages, although adipogenic differentiation capacity was low. Blast transformation reaction demonstrated that these cells significantly suppress T-cell proliferation, and this ability is dose-dependent. Intravenous administration of a cell freezing medium, therapeutic dose of cASCs (2 × 106 live cells/kg), and five times higher dose of cASCs showed no significant side effects in two dogs. Microscopic tissue lesions were limited to only mild, non-specific changes. There were no signs of malignancy. The results of the study indicate that cASCs are similar to hASCs and are safe for therapeutic applications in a canine model. The proposed methodology for ASC preparation on a non-routine basis, which includes individually optimised cell culture conditions and offers risk-adapted treatment, could be used for future personalised off-the-shelf therapies, for example, in myocardial infarction or stroke.

The temporospatial patterns in the localization of hexose transporters as well as in the quantitative and qualitative differences of glycoprotein mucin produced by the goblet cells of broiler chicken (Gallus gallus domesticus) small intestine during their first postnatal month were studied. The integral membrane proteins glucose transporter-2 and -5 (GLUT-2 and GLUT-5) that facilitate the transport of hexoses across epithelial cell layers that separate distinct compartments in organism were detected in the chicken intestinal epithelial cells using immunohistochemical labeling with polyclonal primary antibodies Rabbit anti-GLUT-2 and Rabbit anti-GLUT-5 (IHC kit, Abcam, UK). The chemical composition of mucin (neutral, acid) was carried out by applying the histochemical reactions by Alcian-Blue and periodic acid-Schiff methods. The results revealed presence of the hexose transporters GLUT-2 and -5, immunolocalized in the enterocytes of broiler's small intestine and the temporospatial pattern of the density of goblet cells of intestinal mucosa as well as the chemical composition of mucin produced by the goblet cells in chicken immediately after hatching and in 30-days-old chicken's. Simultanously, when goblet cells remained unstained with both antibodies in intestinal epithelium in chicken of both ages or some moderate staining was noticed in 30-days-old chickens' ileal epithelium, the increase of neutral and acid mucin- containing cells per area unit in both segments of the small intestine was detected from the first day after hatching to 30 day of life and the densilty of goblet cells was found to be higher in ileal than in duodenal region.

Abstract 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.

The role of goblet cell secretion, containing mucopolysaccharides, in the formation of a protective barrier of intestinal mucosa and transportation of the intestinal content has been described quite extensively. However, information on the quality composition of mucopolysaccharides and its changes in the intestinal tract of ostrich chicks, especially in the large intestinal segments, is unavailable. In the current study, ostrich embryos/chicks (n = 6/36) of both sexes were used shortly before hatching and during the first months of the post-hatch period. Tissues for histology were taken from the large intestine: the medium segments of the caecum, proximal and distal parts of colon. By using histochemical reactions, the differentiation of goblet cells as well as chemical composition of mucopolysaccharides was carried out. The cells contained acid (AB+), neutral (PAS+) and mixed (AB/PAS+) mucopolysaccharides. The number of goblet cells in the large intestine per unit area of mucosa increased towards the cloaca, and it was the highest in the distal part of the colon. The qualitative goblet cell composition in different large intestinal parts was different in all ages. In the caecum, goblet cells containing acid and mixed mucopolysaccharides dominate post-hatch, whereas in the colon, goblet cells containing acid mucopolysaccharides predominated. The most rapid changes in the qualitative goblet cell composition occur during the first week post-hatch when in all the intestinal segments the proportion of cells containing acid mucopolysaccharides continuously increased.

The hexoses glucose, galactose and fructose serve as important dietary energy sources in animals and glucose plays a central role in energy homeostasis within eucaryotic cells. As relatively little is known about patterns of hexose transporters expression in birds gastrointestinal tract, the aim of the study was to examine glucose transporters 2 (GLUT-2) and glucose transporters 5 (GLUT-5) expression in stomach and duodenal epithelium of two different species—broilers (Gallus gallus domesticus) and ostriches (Struthio camelus var. domesticus) chicken. Materials from the two parts of gastrointestinal tract were collected from six 7-day-old female broilers and six 7-day-old female ostriches. Specimens were fixed with 10% formalin, embedded into paraffin, cut into 7 μm thick slices, followed by immunohistochemical staining with polyclonal primary antibodies rabbit anti-GLUT-2 and rabbit anti-GLUT-5; the staining was carried out according to the manufacturers guidelines (IHC kit, Abcam, UK). The investigation showed that the staining for both antibodies was more intensive in the epithelial cells of stomach and duodenum of the 7-day-old broilers. In duodenal epithelium, goblet cells and brush border membranes were stained in both species, however the signal was stronger for GLUT-5 than GLUT-2. Staining for GLUT-2 and GLUT-5 occurred in different parts of gastrointestinal tract of 7-day-old ostriches, but was weaker compared to 7-day-old broilers, which showed that the gastrointestinal tract of 7-day-old female broilers was more developed for transportation of hexoses than 7-day-old female ostriches.

The biopsy of live animal testes is an important clinical manipulation to control spermatogenesis and reproductive system pathologies. The aim was to develop a method of boar testes biopsy using a biopsy gun with ultrasound guidance and to investigate the influence of this procedure on the boar testes parenchyma and quality of ejaculate. The biopsy was carried out in six 8-month-old boars. Fourteen days prior to and 21 days after biopsy, the quality of ejaculate was examined (weight of ejaculate; concentration and motility of spermatozoa) with a seven-day intervals. Ultrasound images of the testes parenchyma were recorded three times: directly before and 15 minutes after the biopsy, then 21 days after the procedure. The testes biopsies of generally anesthetized boars were performed with the biopsy gun for needle biopsy with a 12cm long, disposable 16-gauge needle 1.8mm in diameter (Vitesse) through 1cm skin incision in the depth of 1.2-1.6cm of parenchyma. Fifteen minutes after the biopsy, macroscopic injures of the parenchyma of all the boar testes were not detected in the ultrasound image. Twenty one days after biopsy, the hyperechogenic line 0.1-0.2cm in diameter was seen in the testes parenchyma of six boars in the depth of 1.2-1.6cm. The biopsy of boar testes did not influence the quality of boars ejaculate. The ultrasonographic examination of boar testicles before the biopsy reduced possibilities to traumatize large blood vessels of the testes. A perfect boar testicular biopsy was easy to perform using ultrasonographic examination in the pigsty conditions.

Peptides of the gastrointestinal tract play a significant role in the digestive processes and the development of the body; therefore, it is important to have an understanding of location and distribution of gastrin, somatostatin and glucagon immunoreactive (IR) cells in the stomach mucosa of growing birds. For this purpose, 6 embryos and 37 chicks from an ostrich farm in Latvia were used. Tissue samples were collected from the proventriculus - superficial and deep glandular region and from the ventriculus - side wall and pyloric region. The number of cells was determined in 10 mucosal fields of each tissue sample. For statistical analysis, the one-way anova method was used. Gastrin IR cells regarding the stomach mucosa were found only in the pyloric region. Somatostatin IR cells were most densely located in the pyloric region too, but some cells were also discovered in the mucosa of proventriculus and ventriculus. Glucagon IR cells were found in the epithelium of the deep glands of the proventriculus and only some cells of the superficial glands of the proventriculus, and the ventriculus side wall mucosa. Gastrin and somatostatin IR cells were present in a comparatively large quantity in the ostrich chicks' ventriculus - pyloric region yet not long before hatching. They were located deep in the mucosa of pyloric glands, and their number tended to increase with birds advancing in age.

The distribution of the goblet cells has been widely studied both in mammals and in birds, mainly hens; however, the functional studies of cells, especially in growing birds of different species are still topical. The aim of this research was to determine the density of the goblet cells in the mucosa of the ostrich small intestine pre hatch and during the first months of life as well as to differentiate the goblet cells by the chemical composition of mucopolysaccharides. In the research, 42 ostriches of both sexes raised in Latvia were used, including six embryos obtained on the 38th incubation day and 36 chicks of age 1, 3, 7, 14, 30 and 60 days post hatch, distributed in groups of 6 birds in each group. The length of each small intestine segment was measured (mm). Histological samples of tissue (0.5-1x1cm) were taken from the small intestine: the medium segments of the duodenum, jejunum and ileum. For overall histological assessment, the tissue samples were deparaffinized, hydrated and stained with haematoxylin and eosin stain complying the standard methods. By applying the histochemical reactions, the the goblet cells were differentiated by the qualitative composition of mucopolysaccharides into cells containing acid (AB+), neutral (PAS+) and mixed (AB/PAS+) mucopolysaccharides. The density of the goblet cells was determined in 10 villi of each preparation, each segment of the small intestine for each individual. The density of the obtained cells was calculated per 1mm2 of the median longitudinal section of a villus. The data obtained in the study were statistically processed by SPSS 17.5 software programme. The density (number) of the goblet cells of the ostrich small intestine per one area unit of mucosa from the 38th day of embryonic development until the age of 30 days tended to decrease in all segments of the small intestine. On day of hatch, the largest density of the goblet cells per 1 mm2 of mucosa was observed in the duodenum, in turn at 60 days of age in the ileum. Differences of the density of goblet cells and the proportional division were observed depending on the chemical composition of mucopolysaccharides in different segments of the small intestine of ostrich chicks. The obtained results characterize both the quantitative and qualitative differences of mucopolysaccharides (mucus) secretion that is possibly connected with various specific roles of the small intestine segments in the processes of nutrients absorption.

The dynamics of changes of an increase in the ostrich intestinal weight and length in postnatal ontogenesis provides a conception of the feed processing possibilities in some intestinal segments, which, in turn, influences the bird‘s general health condition and production yield. The aim of the research was to find out changes of the small and large intestinal weight and length growth in ostriches from day 120 to day 360 of life. This investigation included 120, 180, 240, and 360 days old 18 ostriches of both sexes, raised in Latvia. The absolute weight and length of the small and large intestine were established. The absolute total weight and length of intestines, the relative weight and length of small and large intestine, and the ratio of small and large intestinal weight to their length were calculated. The obtained data were statistically processed using SPSS 20.0 program. In each age group, the mean arithmetic value and standard error were calculated, and a one-way variance analysis with LSD was used for comparison of data agreement of age groups. The total intestinal weight of ostriches as well as the small and large intestinal absolute weight increased significantly from day 120 to day 240 of life (p<0.05), while further to day 360 there were no marked changes. However, the relative weight of the small and large intestine tended to decrease with ostriches advancing in age. The ratio of the large intestinal weight to that of the small intestine from day 120 to day 360 of life increased linearly from 2.8 to 3.7. The absolute length of ostrich small and large intestine increased significantly from day 120 to day 240 of life (p<0.05). The length of small intestine had a tendency to reduce from 41.8% on day 120 to 36.4% on day 360 of life; whereas the relative length of large intestine at this stage of age tended to grow from 58.2% to 63.6%. The ratio of the large intestinal length to that of the small intestine increased linearly from 1.4 on day 120 to 1.75 on day 360 of life. The total length of intestines increased two times over the research period, but a significant growth was observed from day 120 to day 240 of life (p<0.05).