Effects of induced maternal hypothyroidism on the ovarian development of offspring rats

The effects of propylthyouracil (PTU) induced hypothyroidism of rats during pregnancy and lactation on offspring ovarian development and maturation were studied. Thyroid hormones and thyroid stimulating hormone (TSH) concentrations were determined using the radioimmunoassay method in order to verify the hypothyroid status of treated mothers and their two months old pups. The ovaries of the offspring were processed for light microscopy analysis on the day of the first estrus after the 60th day of age. Histological analysis including follicle count was performed on serial sections stained with haematoxyline/eosin and on semithin sections stained with methylene blue. A significant increase of serum TSH and decrease in T3 and T4 levels was observed in treated mothers compared to controls. The levels of measured hormones in the control and PTU-treated two months old rats were not significantly different. Ten percent of 60-dayold treated females did not reach estrus and they were sacrificed in diestrus. The secondary interstitial cells were the dominant structures in the ovaries. The number of healthy growing and early antral follicles was markedly decreased. Ovaries of treated rats contained relatively few antral follicles, significantly more atretic antral follicles and a decreased number of corpora lutea, compared to controls. These results indicate that lack of thyroid hormones during prenatal and early postnatal development impair ovarian development in rats. [Projekat Ministarstva nauke Republike Srbije, br. 175061

Histological and Immunological Changes in Uterus During the Different Reproductive Stages at Californian Rabbit (Oryctolagus cuniculus)

Rabbit is the third most commonly used animal model in different fields of scientific research, such as reproductive biology, fertility and embryo transfer studies, and immunology. This animal species, often used in antibodies production, has minority of scientific records about the immunological status of its reproductive organs. The aim of this study was to find histological and immunological changes in rabbit female reproductive tract during different reproductive stages. The study was carried out on female rabbits, divided in three groups, according to the following stages of reproductive cycle: Estrous, ovulation and pregnancy. Histological and immunohistochemical stains for T- and B-cells were performed on tissue samples of cornu uteri and cervix. T lymphocytes were predominant in all anatomical parts of the uterus, in all stages of the cycle. The highest number of those cells was recorded at estrous, while the lowest was recorded at pregnancy. Cervix expressed more immunological activity than cornu uteri. The distribution and the number of immune positive cells in the rabbit female reproductive tract depend on its hormonal status

The effects of continual consumption of origanum vulgare on liver transcriptomics

Pathogen control is re-emerging as a significant challenge to the health of both humans and animals. The livestock industry is in the process of massively replacing in-feed antibiotics with organic production friendly plant-based products. Nutrigenomics as a science of the effects of food constituents on gene expression is shedding more light on both benefits and detrimental side-effects of feed additive prolonged consumption on the host, indicating the need to understand the feed– host interactions and their influence on the host disease profile. In this study, we investigated the effects of 2% oregano powder supplementation on the liver gene expression in healthy male broilers from the hatch to 6 weeks of age. Deep RNAseq was performed on average 113.3 million paired and quality trimmed sequences per sample and four samples for the control and treatment each. The results demonstrate the severity of oregano effect on liver gene expression with substantial modifications in steroid hormone regulation, fat and carbohydrate metabolism alterations and strong influence on the host disease and function profile. Oregano supplementation was able to interfere with the transcriptional effects of a range of registered drugs and to significantly transcriptionally inhibit a range of cancer disease categories including liver cancer, and to modify fat and carbohydrate metabolism

Apoptoza kao način prirodnog odumiranja ćelija jajnika

Different hormones, cytokines, the absence of growth factors, and others, are some of the signals for initiating apoptosis in ovarian cells. Each of them in its own way, trigger apoptosis as a form of death in which the cell actively participates by precisely implementing a genetically programmed sequence of biochemical and morphological changes which lead to selfdestruction. Apoptosis is a physiological form of death, which helps establish a dynamic balance among proiliferation, differenciation, and death of ovarian cells. It has been confirmed so far that follicular cells oocytes, cells of the germinal epithelium, theca cells, and corpus luteum cells die through apoptosis. The physiological deaths of these cells are an integral part of normal ovarian function, both during intrauterine and postnatal life. Namely, during intrauterine ovarian development, about half the total number of germinative cells (future oocytes) die through apoptosis and their population is gradually reduced after birth by so-called selection of follicles which will continue further growth (folliculogenesis) and the apoptosis of cells of those follicles which will be subjected to atresion. Most ovarian cells die by apoptosis continuously until the end of the reproductive life period of healthy females, and some can continue dieing in this way until the death of the given individual (e.g. germinal epithelium cells).Među signale za pokretanje apoptoze u ćelijama jajnika ubrajaju se razni hormoni, citokini, odsustvo faktora rasta i drugi. Svaki od njih, na svoj način, pokreće apoptozu kao oblik smrti u kome ćelija aktivno učestvuje tako što precizno sprovodi genetski programiran sled biohemijskih i morfoloških promena koje je vode u autodestrukciju. Apoptoza je fiziološki oblik smrti pomoću kojeg se obezbeđuje uspostavljanje dinamičke ravnoteže između proliferacije, diferencijacije i odumiranja ćelija jajnika. Do sada je potvrđeno da u jajnicima apoptozom odumiru folikularne ćelije, ovociti ćelije klicinog epitela, ćelije teke i žutog tela. Fiziološka smrt ovih ćelija je sastavni deo normalne funkcije jajnika, kako tokom intrauterinog tako i za vreme postnatalnog života. Naime, tokom intrauterinog razvoja jajnika apoptozom odumre oko polovina od ukupnog broja germinativnih ćelija (budućih ovocita), a posle rođenja njihova populacija postepeno se smanjuje "odabirom folikula" koji će nastaviti dalji razvoj (folikulogeneza) i apoptotskim odumiranjem ćelija onih folikula koji će podleći atreziji. Većina ćelija jajnika odumire apoptozom kontinuirano do kraja reproduktivnog perioda života zdravih ženki, a neke mogu da odumiru, na ovaj način, sve do smrti jedinke (na primer ćelije klicinog epitela)

Efekat kratkotrajnog gladovanja na koncentraciju lipida i lipoproteina kod zdravih pasa normalne telesne kondicije

Introduction. Analysis of canine lipoprotein fractions after agarose gel electrophoresis (agEF) separation could be an important diagnostic tool in primary and secondary dyslipidemia diagnosis. The aim of this study was to measure concentrations of triglycerides and cholesterol and to analyze lipoprotein fractions in dogs after basal (12 hours) and short-term (24 and 36 hours) fasting, i.e., frequent conditions in clinical practice. Materials and Methods. Blood samples were collected from six lean dogs of both sexes and different breeds, after 12, 24 and 36 hours of fasting. Concentrations of glucose, triglycerides and cholesterol were determined on an automated wet biochemistry analyzer, lipoprotein fractions were separated by agEF and leukocyte numbers were assessed on an automated hematology analyzer. Results and Conclusions. Results showed there was no significant change in glucose, triglyceride and cholesterol concentrations nor in leukocyte numbers during dog fasting. Moreover, there was no change in α1- and α2-fractions, but there was a significant decrease in preβ- and β-fraction of lipoproteins. It is know that high density lipoproteins (HDL) have α-mobility and very low density (VLDL) and low density lipoproteins (LDL) have preβ- and β-mobility. Thus, it is possible that reverse cholesterol transport maintained by HDL is not affected during short-term fasting. On the contrary, synthesis of VLDL and formation of LDL are probably decreased because endogenous synthesis of triglycerides is decreased or their clearance is increased.Uvod. Poznato je iz literature da se odnosi lipoproteinskih frakcija seruma pasa koje se dobijaju posle elektroforeze na agaroznom gelu (EFag) menjaju kod oboljenja praćenih dislipidemijama, što može biti jedan od značajnih elemenata dijagnostike pojedinih patoloških stanja. Stoga je cilj ovih ispitivanja bio da se utvrdi koncentracija triglicerida, holesterola i glukoze u serumu pasa kao i odnos lipoproteinskih frakcija dobijenih pomoću EFag, u uslovima koji se često sreću u kliničkoj praksi: posle bazalnog (12 časova) i kratkotrajnog gladovanja od 24 i 36 časova. Materijal i metode. Uzorci krvi su sakupljeni od šest pasa različitih rasa, normalne telesne kondicije, oba pola, posle 12, 24 i 36 časova gladovanja. Koncentracija glukoze, triglicerida i holesterola je određena na automatskom biohemijskom analizatoru. Lipoproteinske frakcije su razdvojene korišćenjem EFag. Broj leukocita je utvrđen na hematološkom analizatoru. Rezultati i zaključak. Rezultati ovog ispitivanja su pokazali da se tokom kratkotrajnog gladovanja kod pasa koncentracija glukoze, triglicerida i holesterola kao i broj leukocita ne menjaju. Osim toga, nije bilo promena vezanih za α1-i α2-elektroforetske zone, ali se javio značajan pad koncentracije lipoproteina preβ-i β-zone. Poznato je da se lipoproteini velike gustine (HDL) nalaze u α-zoni, dok lipoproteini veoma male gustine (VLDL) i lipoproteini male gustine (LDL) pripadaju preβ-i β-zoni. Na osnovu naših nalaza moguće je zaključiti da reverzni transport holesterola koji se odvija posredstvom HDL-a nije poremećen tokom kratkotrajnog gladovanja. Nasuprot tome, kratkotrajno gladovanje verovatno utiče na smanjen obim sinteze triglicerida u jetri, odnosno na smanjenu sintezu VLDL-a i LDL-a ili na njihovo efikasnije uklanjanje iz plazme

Stereological and immunohistochemical study of the spleen in hypothyroid juvenile rats

The purpose of this work was to investigate the influence of hypothyroidism on spleen tissue morphology and immune cell density in fourteen-day-old juvenile rats. Hypothyroidism in pups (n=10) was induced by administration of propylthiouracil (PTU) in drinking water (1.5 mg/L) to their mothers during pregnancy and period of lactation. Fourteen-day-old pups were sacrificed and the thyroid-stimulating hormone (TSH) serum concentration and thyroid activation index (Ia) were determined. Increased serum level of TSH and increased Ia showed that pups from PTU treated mothers were hypothyroid. White and red spleen pulp, marginal zone and connective tissue volume density has been assessed by using the stereological method. Using immunohistochemistry, the present CD3+ T lymphocytes, CD45RA+ B lymphocytes and CD68+ macrophages were quantified. A significant reduction of volume density of the periarteriolar lymphocyte sheath (Vv(PALS)) and lymphatic follicles (Vvf) due to depletion of T and B lymphocytes respectively, was observed in the spleens of hypothyroid pups compared to controls. The volume density of the red pulp (Vvrp), marginal zone (Vvmz) and connective tissue (Vvct) was increased, as well as the number of CD68+ macrophages in the spleens of hypothyroid pups compared to controls. These results indicate that thyroid hormones might be important for normal development of both, specific and innate immune cells in the spleen during prenatal and early postnatal period

The effects of immunocastration on male pig yield parameters and meat quality

The aim of this study was to evaluate the effects of immunocastration on pork meat and carcass quality, compared to meat from surgically castrated males and entire males. Ninety (Duroc x Pietrain) x (Landrace x Yorkshire) crossbred pigs were assigned to three experimental groups: surgically castrated males (barrows; castrated up to the seventh day of age), entire males (males), and vaccinated males (immunocastrates). Carcass and meat quality characteristics such as weight of hot and chilled carcass, meat yield, chilling loss and chemical parameters were examined. Surgically castrated pigs had significantly lower (p lt 0.01; p lt 0.05) weight before slaughter, than males and immunocastrates, and also lower (p lt 0.01) warm carcass weight than males. The average carcass meatiness of castrates was significantly lower (p lt 0.01) than the average meatiness of males and immunocastrates. Chilling loss of barrows was significantly lower (p lt 0.01) than chilling loss of males or immunocastrates. It was also found that the chilling loss of immunocastrates was significantly lower (p lt 0.05) than chilling loss of males. According to the results obtained, it can be concluded that immunocastration could be a good alternative to surgical castration considering meat and carcass quality characteristics

Površinski epitel jajnika hipotireoidnih novorođenih i neonatalnih pacova - iz perspektive PCNA i kaspaze-3

Introduction. The ovarian surface epithelium (OSE) undergoes intensive regeneration and remodelling after each ovulation during the whole reproductive period. This process increases the risk of one of the most common ovarian tumors in women and the female dog. Considering the fact that maternal hypothyroidism highly impacts cell proliferation and cell death during folliculogenesis in the early neonatal period, we aimed to analyze its effect on OSE morphology and dynamics. Materials and Methods. The study was performed on newborn (24-h-old) and neonatal (4-day-old) female rats, a randomized trial between the control and hypothyroid groups, born under controlled circumstances and hypothyroid mothers, respectively. Their ovaries were analyzed histologically and processed to determine the OSE cell height as an average value of four measurement points. Also, the immunopositivity of the proliferating cell nuclear antigen (PCNA) and caspase-3 were assessed semiquantitatively. Results and Conclusions. No major structural differences of OSE were found between groups within the given ages except for a slight increment of OSE cell height and incompleteness of apical cell membrane with cytoplasmic projections in hypothyroid animals. PCNA immunopositivity of the OSE cells was higher in ovaries of hypothyroid animals of both ages in comparison to the controls. Moreover, only scarce OSE cells were caspase-3 positive in both groups and ages, with no difference in immunopositivity. Our study confirms the impact of hypothyroidism in the early postnatal period on morphology and proliferation rate of OSE cells, with no effect on caspase-3 dependent cell removal, which may serve as a premise for future investigation of potential carcinogenesis, in terms of prevention and treatment of ovarian cancer.Uvod. Tokom reproduktivnog perioda, nakon svake ovulacije, površinski epitel (PE) jajnika se intenzivno regeneriše i remodelira. Ovaj proces povećava rizik od nastajanja jednog od najčešćih tumora jajnika kod žena i kuja. Imajući u vidu činjenicu da hipotireoidizam majki u velikoj meri utiče na proliferaciju i ćelijsku smrt u toku folikulogeneze u ranom postnatalnom periodu, cilj rada je bio da analiziramo efekat hipotireoidizma majki na morfologiju i dinamiku PE jajnika potomaka. Materijal i metode. U eksperimentu su korišćeni tek rođeni (24 časa stari) i neonatalni (4 dana stari) mladunci koji potiču od kontrolnih i hipotireoidnih majki. Morfometrijskom analizom određivana je prosečna visina ćelija PE, izmerena na četiri mesta na svakom ispitivanom preseku jajnika. Takođe, imunopozitivnost proliferativnivnog ćelijskog nuklearnog antigena (PCNA) i kaspaze 3 je određivana semikvantitativno. Rezultati i zaključci. Veće promene u strukturi PE jajnika nisu zapažene između grupa, osim blagog povećanja visine ćelija i nepotpune apikalne membrane sa projekcijama citoplazme kod hipotireoidnih životinja. Imunopozitivnost PCNA je značajno povećana u jajnicima hipotireoidnih životinja obe starosne grupe u odnosu na kontrole, dok je imunopozitivnost na kaspazu 3 slabo izražena kod svih životinja. Naše istraživanje je pokazalo da hipotireoidizam u ranom postnatalnom periodu dovodi do promena u morfologiji i proliferaciji ćelija PE, bez efekta na uklanjanje ćelija delovanjem kaspaze 3. Ovaj model može da posluži u budućim istraživanjima potencijalne karcinogeneze u cilju prevencije i tretmana kancera jajnika

Z-cells and oogonia/oocytes in the advanced process of autophagy are the dominant altered cells in the ovaries of hypothyroid newborn rats

Induced prenatal hypothyroidism in rat pups leads to accelerated primordial follicle assembly and premature follicular atresia with ovary failure. This work investigates the influence of maternal hypothyroidism induced with 6-n-propyl-2-thyouracil (PTU) on the number and morphology of oogonia/oocytes in newborn rat pups with light and transmission electron microscopy. Expression of apoptosis and autophagy markers in oogonia/oocytes were examined using immunohistochemistry. Hypothyroid newborn pups had a decreased number of mitotic and resting oogonia, while the number of altered oogonia/oocytes was increased. Ultrastructural observations revealed the increased presence of degenerated pachytene oocytes (Z-cells) and oogonia/oocytes undergoing autophagy, apoptosis and combined apoptosis and autophagy, in this group. The most abundant altered oogonia/oocytes in the hypothyroid group were those with morphological features of advanced autophagy and Z-cells. The percentage of TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) positive oogonia/ oocytes was significantly lower in the hypothyroid group. No significant difference was recorded in the expression of caspase-3, ATG7 and LC3 possibly reflecting that these proteins were not involved in the oogonia/ oocyte alteration process during prenatal rat hypothyroidism. The obtained results indicate that developmental hypothyroidism in the offspring enhances the number of Z-cells and oogonia/ oocytes altered with the advanced process of autophagy

Uticaj energetskog dodatka u ishrani na koncentraciju tireoidnih hormona, insulinu sličnog faktora rasta-i i njegovih vezujućih proteina u krvi krava tokom rane laktacije

The objective of this study was to examine the effect of dietary energy supplementation on hormones that are considered to be the main signals of a shift in energy balance around parturition. Sixty dry cows, 15 days before calving, were chosen and divided into two eaqual groups: control and experimental (GLY). Both groups were fed a standard ration balanced in accordance to the stage of the productive reproductive cycle. Additionally, each cow in the GLY group was given glycerol based dietary energy supplementation (250 mL daily during the dry and 300 mL daily during the lactation period), which provided additional 9.30 MJ NEL during the dry and 13.95 MJ NEL during the early lactation period. Milk production was measured on days 30 and 60 of lactation and milk production was significantly higher in GLY compared to control group at day 60 of lactation (p lt 0.05). Service period and insemination index were used as reproductive outcome parameters. Average service period in the control group was significantly longer than in the GLY group (p lt 0.05). Average insemination index in the control group was not significantly different than the index obtained for the GLY group. Blood samples were taken before the begining of the experiment (15 days before parturition), and at days 7, 30 and 60 of lactation. Concentrations of thyroid hormones, IGF-I, relative abundance of IGFBP-2, IGFBP-3 and IGFBP-4, concentrations of total protein and albumin in the blood were measured. Results showed that at days 7 and 30 after parturition, T4 concentrations were significantly higher (p lt 0.001, respectively) in GLY than in the control group, while T3 concentrations were significantly higher in GLY group only at day 7 after parturition (p lt 0.001). IGF-I concentrations and IGFBP-3 abundance were significantly higher in the GLY compared to the control group in all three examined postpartum periods. IGFBP-2 and IGFBP-4 concentrations were higher in GLY compared to the control group in all three examined postpartum periods, but the difference was significant only on day 60 after parturition (p lt 0.01, respectively). Concentrations of total protein and albumin were significantly higher in GLY compared to the control group in all three examined periods after parturition. Based on these results it can be concluded that peripartum dietary energy supplementation prevent cows' exposure to severe negative energy balance, preserves synthetic activity of hepatocytes and consequently has a positive impact on milk production and reproductive performances in dairy cows.Cilj ovog rada je bio da se ispita uticaj energetskog dodatka u ishrani krava na koncentraciju hormona u krvi koji su u periodu oko teljenja glavni pokazatelji promena u energetskom statusu. U tu svrhu, petnaest dana pre teljenja odabrano je 60 krava koje su podeljene u dve jednake grupe: kontrolnu i oglednu (GLY). Obe grupe krava su dobijale identičan obrok usklađen sa proizvodno reproduktivnim ciklusom. Dodatno, kravama GLY grupe je tokom poslednje dve nedelje zasušenja i do 60. dana laktacije dodavan energetski dodatak na bazi glicerola (250 mL dnevno tokom zasušenja odnosno 300 mL nakon teljenja), obezbeđujući dodatnih 9,30 MJ NEL tokom zasušenja, odnosno 13,95 MJ NEL tokom rane laktacije. Kod svih krava je izmerena prosečna dnevna proizvodnja mleka 30. i 60. dana lakatcije, koja je kod GLY grupe bila značajno viša 60. dana (p lt 0,05). Kao pokazatelji reproduktivnog statusa koriščeni su servis period i indeks osemenjavanja. Servis period je bio značajno duži kod kontrolne u odnosu na GLY grupu (p lt 0,05), a vrednost indeksa osemenjavanja se nije značajno razlikovala između grupa. Uzorci krvi krava uzeti su neposredno pre početka ogleda, odnosno 15 dana pre teljenja, kao i 7, 30. i 60. dana laktacije i u njima je određivana koncentracija tireoidnih hormona, IGF-I, relativna zastupljenost IGFBP-2, IGFBP-3 i IGFBP-4, koncentracija ukupnih proteina i albumina. Rezultati su ukazali da je 7. i 30. dana nakon teljenja GLY grupa imala značajno višu koncentraciju T4 (p lt 0,001, pojedinačno) u odnosu na kontrolu, dok je koncentarcija T3 bila značajno viša kod GLY grupe 7. dana nakon teljenja (p lt 0,001). Koncentracija IGF-I i zastupljenost IGFBP-3 je bila značajno viša u krvi krava GLY grupe u odnosu na kontrolu u sva tri ispitivana perioda posle teljenja. Zastupljenost IGFBP-2 i IGFBP-4 je bila viša u krvi krava GLY u odnosu na kontrolnu grupu u sva tri ispitivana perioda postapartalno ali je ova razlika bila značajna jedino 60. dana lakatcije (p lt 0.01, pojedinačno). Koncentracija ukupnih proteina i albumina je, takođe, bila značajno viša u krvi krava GLY grupe u odnosu na kontrolnu grupu u sva tri ispitivana perioda posle teljenja. Na osnovu dobijenih rezultata može se zaključiti da energetski dodatak u ishrani sprečava izloženost krava izrazitom negativnom energetskom bilansu, održava sintetsku funkciju hepatocita i posledično ima pozitivan uticaj na mlečnost i reproduktivne pokazatelje