Synergistic effect of gibberellin and light on the induction of somatic embryogenesis from lateral roots of spinach (Spinacia oleracea L.) in vitro

U ovom radu analiziran je uticaj giberelina (GA) i svetlosti na indukciju somatske embriogeneze iz apikalnih odsečaka korenova spanaća sorte „Matador“. Izolovani organi kultivisani su u uslovima in vitro na medijumu za indukciju somatske embriogeneze, koji je sadržao 20 μM α-naftil-sirćetnu kiselinu (NAA) i 5 μM giberelnu kiselinu (GA3). Velika varijabilnost u embriogenom potencijalu spanaća, koja je posebno izražena kod sorte „Matador“, otežava proučavanje delovanja drugih faktora na proces somatske embriogeneze. Analizom embriogenog potencijala nasumično izabranih klijanaca dobijenih isklijavanjem semena sorte “Matador” devet različitih evropskih proizvođača semena, uočeno je značajno variranje frekvence regeneracije (0-100%) između populacija i individua u okviru populacije, kao i prosečnog broja somatskih embriona (SE) po eksplantatu (0-14.4 po populaciji, odnosno 0-36 po eksplantatu pojedinačnih klijanaca). Rezultati ukazuju na važnost odabira biljnog materijala za eksperiment, jer on može značajno uticati na interpretaciju rezultata. Stoga je u ovom radu razvijena i uspešno korišćena metoda uzorkovanja biljnog materijala, izlaganjem jednakog broja odsečaka korenova svakog klijanca svim tretmanima u okviru eksperimenta, čime je prevaziđen uticaj varijabilnosti biljnog materijala usled genetičkih i drugih uticaja. U cilju ispitivanja mehanizma kojim GA3 indukuje proces somatske embriogeneze, testiran je uticaj paklobutrazola (PAC), inhibitora biosinteze GA i trihostatina A (TSA), inhibitora deacetilaze histona, na ovaj proces. Suprotno očekivanju, PAC je delovao sinergistički sa GA3 i NAA u stimulaciji somatske embriogeneze. U kombinaciji sa NAA+GA3, 1–10 M PAC je uticao na povećanje embriogenog potencijala, sa najvećim efektom pri 2.5 μM PAC. U kombinaciji sa NAA, koji sam po sebi nije dovoljan za indukciju somatske embriogeneze, bila je potrebna nešto viša koncentracija PAC za dostizanje značajnog embriogenog odgovora—10 M PAC za eksplantate nasumično izabranih klijanaca i 5 M PAC za eksplantate genetički predisponiranih jedinki prema procesu somatske embriogeneze. TSA (0.1–5 μM primenjen u trajanju od 1, 7 ili 14 dana) nije mogao da zameni ni NAA ni GA3 u indukciji somatske embriogeneze, ali je u kombinaciji sa 10 μM NAA+5 μM GA3 ili 20 μM NAA+5 μM GA3 uticao na značajno povećanje embriogenog potencijala eksplantata. Pozitivan uticaj TSA na indukciju somatske embriogeneze je pre svega zavisio od dužine trajanja tretmana. Duži TSA treatman je bio potreban u kombinaciji sa nižom koncentracijom NAA i obrnuto. Uočen je i značajan uticaj dimetil sulfoksida, koji je korišćen kao rastvarač TSA, na povećanje embriogenog potencijala eksplantata. Kvalitet svetlosti je takođe značajno uticao na embriogeni potencijal eksplantata. Najveći embriogeni potencijal su imali eksplantati kultivisani pod plavom LED svetlošću. Eksplantati kultivisani pod belom LED i fluorescentnom svetlošću su imali značajno niži embriogeni potencijal, dok je regeneracija somatskih embriona bila potpuno inhibirana u eksplantatima kultivisanim pod crvenom LED svetlošću ili u mraku. Da bi se rasvetlio mehanizam delovanja GA i svetlosti na indukciju procesa somatske embriogeneze, sprovedena je detaljna analiza metabolizma GA u eksplantatima tokom indukcije somatske embriogeneze analizom ekspresije gena koji kodiraju ključne enzime za biosintezu (SoGA20-ox1 i SoGA3-ox1) i inaktivaciju (SoGA2-ox1, SoGA2-ox2 i SoGA2-ox3) GA, kao i sadržaja endogenih GA. Najveće razlike između embriogenih eksplantata (kultivisanih na medijumu sa NAA+GA3), i neembriogenih eksplantata (kultivisanih na medijumu bez biljnih regulatora rastenja i razvića ili na medijumu sa GA3 ili NAA) su detektovane u ekspresiji SoGA20-ox1 i SoGA2-ox2 gena...In this work, the impact of gibberellins (GA) and light, on somatic embryo initiation in vitro from root apices of spinach seedlings (cv. Matador) was studied. The explants were cultivated on induction medium supplemented with 20μM α-naphthaleneacetic acid (NAA) and 5 μM gibberellic acid (GA3). A high variability in embryogenic capacity of spinach, especially in the cultivar "Matador", impedes studying the effects of other factors on the induction of somatic embryogenesis. Analysis of embryogenic potential of randomly chosen seedlings, obtained from seeds of cv. “Matador” purchased from nine European seed companies, showed significant variation in the regeneration frequency (0-100%) among populations and individuals within the population, as well as in the mean SE number per explant (0-14.4 and 0-36 per population and per individual seedlings' explants, respectively). The results strongly evidenced the importance of the sampling of plant material, because it may significantly affect the interpretation of the data. With the aim to overcome such high variability of plant material due to genetic and other impacts, a sampling method was developed and successfully applied in the present work. The sampling method implies the exposure of equal number of root explants taken from each seedling to all treatments. In order to investigate the mechanism of GA3 action on somatic embryogenesis induction, the influence of paclobutrazole (PAC), an inhibitor of GA biosynthesis, and trichostatin A (TSA), a histone deacetylase inhibitor, on this process was studied. Unexpectedly, PAC acted synergistically with GA3 and NAA in promoting somatic embryogenesis. In combination with NAA+GA3, 1–10 μM PAC increased embryogenic capacity of the explants, with maximum effect at 2.5 μM PAC. In combination with NAA, which alone is not sufficient to induce somatic embryogenesis, higher concentrations of PAC were required for satisfactory embryogenic response—10 μM PAC for the explants taken from randomly chosen seedlings and 5 μM PAC for the explants derived from the seedlings genetically predisposed for somatic embryogenesis. TSA (0.1–5 μM applied over 1, 7 or 14 days) could not substitute for either NAA or GA3 in somatic embryogenesis induction, although it significantly increased embryogenic capacity of the explants in combination with 10 μM NAA+5 μM GA3 or 20 μM NAA+5 μM GA3. The positive impact of TSA on somatic embryogenesis induction mostly depended on the treatment duration. Longer TSA treatments were required with lower level of NAA and vice versa. In addition, a significant positive impact of dimethyl sulfoxide, used to dissolve TSA, on embryogenic capacity of the explants was observed. The quality of the light also significantly affected the embryogenic potential of the explants. Explants cultured under blue LED light exhibited the highest embryogenic potential. Explants cultured under white LED and fluorescent light had significantly lower embryogenic potential, while somatic embryo regeneration was completely inhibited in explants cultured under red LED light or in the dark. To elucidate the mechanism of action of GA and light on somatic embryogenesis induction, a detailed analysis of GA metabolism in the explants during induction of somatic embryogenesis was performed by analyzing the expression of genes encoding key enzymes in GA biosynthesis (SoGA20-ox1 and SoGA3-ox1) and inactivation (SoGA2-ox1, SoGA2-ox2 and SoGA2-ox3), as well as the endogenous GA content..

Relation between erythrocyte parameters and stillbirth in piglets

The aim of this study was to investigate the relation between erythrocyte parameters of piglets and their mortality during parturition. Erythrocyte count, haematocrit volume, blood haemoglobin concentration, mean cell volume (MCV), mean cell haemoglobin (MCH) and mean cell haemoglobin concentration (MCHC) were determined in 80 live-born and 109 stillborn piglets. In both live-born and stillborn piglets the investigated parameters were evaluated with respect to sex and birth mass. Blood samples of all piglets were collected immediately after birth. Stillborn piglets had lower erythrocyte values (P<0.01), haemoglobin (P<0.05) and haematocrit (P<0.01) than live-born piglets. The results suggest that the incidence of stillbirth in piglets is associated with reduced levels of erythrocytes and haemoglobin in their blood. There are no significant influences of sex and birth mass on any of the investigated erythrocyte parameters in either live-born or stillborn piglets

Histopatološke promjene u sirištima jelena lopatara na Brijunima - kratko priopćenje.

This study was performed to establish histopathological changes in the abomasa of fallow deer (Dama dama) from the Brijuni islands, in order to investigate the possible presence of Helicobacter species in this species, since literature data on abomasal histopathology are scarce, and data on the presence of Helicobacter species in red deer are absent. The study was performed on 23 abomasal samples taken from young adults (c. 1 year of age) shot in legal hunts. In all the samples inflammatory changes were visible. The histopathological findings from the samples examined were similar, indicating the epizootical nature of the disease, and they were similar to findings described in dogs with a gastric infection from Helicobacter-like organisms. However, in the abomasa of fallow deer Helicobacter organisms were not detected after the appropriate stains were applied, therefore the aetiological factor responsible for such histopathological changes has yet to be identified.Istraživanje je provedeno da bi se ustanovile patohistološke promjene u sirištima jelena lopatara (Dama dama) s brijunskog otočja i eventualna prisutnost Helicobacter vrsta u ove vrste, budući da je malo literaturnih podataka koji se tiču patohistoloških promjena u sirištu. Pregledom literature nismo pronašli niti jednu referencu o prisutnosti Helicobacter vrsta u jelena lopatara. Istraživanje je provedeno na 23 uzorka sirišta mladih odraslih jedinki (starosti oko godinu dana), žrtvovanih u redovitom odstrelu. U svim uzorcima ustanovljene su upalne promjene u sirištu. Patohistološki nalazi svih pregledanih uzoraka bili su slični, što ukazuje na epizootsku prirodu bolesti, te sličnost nalazima u želucima pasa inficiranih helikobakteru sličnim organizmima. Međutim, u sirištima jelena lopatara nakon obrade odgovarajućim tehnikama bojenja nismo ustanovili Helicobacter vrste, te zaključujemo da je etiološki čimbenik odgovoran za promjene nepoznat

Characterization of soil for production of cabbage

This paper reviews a part of analyses of soils used for production of cabbage in the production fields around the village of Futog near Novi Sad. The analyzed soils are heterogeneous but invariably of light mechanical composition. The topsoils have unfavorable water/air properties, low total porosity (from 36.82% to 42.70%) and medium water permeability (from 10-3 to 10-5 cm/s). Low humus content (from 1.62% to 2.17%), highly variable phosphorus content (from 6.5 to 107.0 mg 100-1 g of soil) and medium to high potassium content (from 15.5 to 70.0 mg 100-1 g of soil) are common properties of the surface soil layers of all soil profiles studied. Based on the proportion and biodiversity of the studied groups of microorganisms and dehydrogenase activity, it has been concluded that the analyzed soils have a high biological value

Reptile haematology

Hematologija gmazova je područje koje se u posljednjih nekoliko desetljeća počelo razvijati i istraživati zbog sve većeg broja gmazova koji se drže kao kućni ljubimci, a time su i češći pacijenti u veterinarskim ambulantama. Izrada krvne slike gmazova je postupak koji se za sada još ne radi u kliničkoj veterinarskoj praksi, ali upoznavanjem hematologije i načinima određivanja hematoloških parametara u gmazova otvara se mogućnost i veterinarima male prakse da rutinski odrede hemogram gmazova. Svi se hematološki parametri određuju ručno pa je za dobivanje kompletne krvne slike, osim stručnosti i iskustva osobe koja izrađuje hemogram, potrebna i odgovarajuća laboratorijska oprema i reagensi. Sve krvne stanice gmazova imaju jezgru pa je tijekom diferencijacije stanica potreban oprez kako ne bi došlo do zamjene pojedinih vrsta stanica. Na broj i morfologiju krvnih stanica gmazova utječu brojni vanjski i unutarnji čimbenici koje treba imati na umu tijekom interpretacije dobivenih nalaza. Promjena broja i morfologije krvnih stanica tijekom različitih bolesti i patoloških stanja javlja se ovisno o vrsti gmazova, težini i dužini trajanja bolesti.Reptile haematology has begun increase in the number of reptiles held as developing in recent decades with the household pets and presenting as veterinary patients. Although complete blood count is not presently a routine test in clinical veterinary practice, getting to know the haematology and the means of determining haematological parameters in reptiles will enable small animal practitioners to develop this test as a standard and routine laboratory test. All haematological parameters are to be performed manually, therefore adequate laboratory equipment, chemical reagents and expertise and experience of the veterinarian is mandatory. All reptile blood cells contain a nucleus, so caution is needed in the determination of blood cell types. The number and morphology of blood cells in reptiles is influenced by numerous external and internal factors, which need to be considered during the interpretation of results. Changes in the number and morphology of blood cells during disease and pathological conditions depend on species, body mass and duration of the disease

Reptile haematology

Hematologija gmazova je područje koje se u posljednjih nekoliko desetljeća počelo razvijati i istraživati zbog sve većeg broja gmazova koji se drže kao kućni ljubimci, a time su i češći pacijenti u veterinarskim ambulantama. Izrada krvne slike gmazova je postupak koji se za sada još ne radi u kliničkoj veterinarskoj praksi, ali upoznavanjem hematologije i načinima određivanja hematoloških parametara u gmazova otvara se mogućnost i veterinarima male prakse da rutinski odrede hemogram gmazova. Svi se hematološki parametri određuju ručno pa je za dobivanje kompletne krvne slike, osim stručnosti i iskustva osobe koja izrađuje hemogram, potrebna i odgovarajuća laboratorijska oprema i reagensi. Sve krvne stanice gmazova imaju jezgru pa je tijekom diferencijacije stanica potreban oprez kako ne bi došlo do zamjene pojedinih vrsta stanica. Na broj i morfologiju krvnih stanica gmazova utječu brojni vanjski i unutarnji čimbenici koje treba imati na umu tijekom interpretacije dobivenih nalaza. Promjena broja i morfologije krvnih stanica tijekom različitih bolesti i patoloških stanja javlja se ovisno o vrsti gmazova, težini i dužini trajanja bolesti.Reptile haematology has begun increase in the number of reptiles held as developing in recent decades with the household pets and presenting as veterinary patients. Although complete blood count is not presently a routine test in clinical veterinary practice, getting to know the haematology and the means of determining haematological parameters in reptiles will enable small animal practitioners to develop this test as a standard and routine laboratory test. All haematological parameters are to be performed manually, therefore adequate laboratory equipment, chemical reagents and expertise and experience of the veterinarian is mandatory. All reptile blood cells contain a nucleus, so caution is needed in the determination of blood cell types. The number and morphology of blood cells in reptiles is influenced by numerous external and internal factors, which need to be considered during the interpretation of results. Changes in the number and morphology of blood cells during disease and pathological conditions depend on species, body mass and duration of the disease

Koštana alkalna fosfataza, osteokalcin i C-terminalni telopeptid kao pokazatelji koštane pregradnje u kuja

In clinical veterinary practice the measurement of biochemical markers for assessment of bone metabolism is not yet fully established and is only being used for now as a scientifi c method in investigations. The purpose of this research is the better understanding of bone metabolism in dogs and assessing the possible utility of biochemical bone markers in routine clinical veterinary practice. The bone markers selected for this study were: bone alkaline phosphatase and osteocalcin as indicators of bone formation, and C-terminal telopeptide of type I collagen as marker of bone resorption. The research was conducted on 58 canine bitches of different breeds. The results indicate increased bone turnover in young animals and decreased turnover in old ones since the highest activities of the investigated bone markers were in the youngest group and they then declined with age. However, elevated activity of bone alkaline phosphatase was noted in older bitches, which has not been detected in other animal species or in healthy humans. Higher bone resorption and lower formation were noticed in heavier dogs.U veterinarskoj medicini saznanja o biokemijskim pokazateljima koštane pregradnje oskudna su i nedostatna, a njihova primjena je za sada isključivo istraživačka. Cilj ovog istraživanja bio je bolje poznavanje koštanoga metabolizma u pasa na temelju mjerenja biokemijskih pokazatelja koštane pregradnje u krvi te procjena moguće kliničke primjene tih pokazatelja u rutinskoj veterinarskoj kliničkoj praksi. Pokazatelji koštane pregradnje koji su upotrijebljeni u ovom istraživanju jesu: koštana alkalna fosfataza i osteokalcin, kao pokazatelji izgradnje i C-terminalni telopeptid kao pokazatelj razgradnje kosti. Istraživanje je provedeno na 58 kuja različitih pasmina. Rezultati pokazuju pojačanu pregradnju kostiju u mladih životinja i smanjenu u starih životinja budući da su najviše vrijednosti koštanih pokazatelja utvrđene u mladih životinja, te su opadale s dobi. Međutim, povećana aktivnost koštane alkalne fosfataze zabilježena je u starijih kuja, što nije zapaženo u ostalih životinjskih vrsta niti zdravih ljudi. Također je uočena povećana razgradnja i smanjena izgradnja kosti u pasa veće tjelesne mase

Koštana alkalna fosfataza, osteokalcin i C-terminalni telopeptid kao pokazatelji koštane pregradnje u kuja

In clinical veterinary practice the measurement of biochemical markers for assessment of bone metabolism is not yet fully established and is only being used for now as a scientifi c method in investigations. The purpose of this research is the better understanding of bone metabolism in dogs and assessing the possible utility of biochemical bone markers in routine clinical veterinary practice. The bone markers selected for this study were: bone alkaline phosphatase and osteocalcin as indicators of bone formation, and C-terminal telopeptide of type I collagen as marker of bone resorption. The research was conducted on 58 canine bitches of different breeds. The results indicate increased bone turnover in young animals and decreased turnover in old ones since the highest activities of the investigated bone markers were in the youngest group and they then declined with age. However, elevated activity of bone alkaline phosphatase was noted in older bitches, which has not been detected in other animal species or in healthy humans. Higher bone resorption and lower formation were noticed in heavier dogs.U veterinarskoj medicini saznanja o biokemijskim pokazateljima koštane pregradnje oskudna su i nedostatna, a njihova primjena je za sada isključivo istraživačka. Cilj ovog istraživanja bio je bolje poznavanje koštanoga metabolizma u pasa na temelju mjerenja biokemijskih pokazatelja koštane pregradnje u krvi te procjena moguće kliničke primjene tih pokazatelja u rutinskoj veterinarskoj kliničkoj praksi. Pokazatelji koštane pregradnje koji su upotrijebljeni u ovom istraživanju jesu: koštana alkalna fosfataza i osteokalcin, kao pokazatelji izgradnje i C-terminalni telopeptid kao pokazatelj razgradnje kosti. Istraživanje je provedeno na 58 kuja različitih pasmina. Rezultati pokazuju pojačanu pregradnju kostiju u mladih životinja i smanjenu u starih životinja budući da su najviše vrijednosti koštanih pokazatelja utvrđene u mladih životinja, te su opadale s dobi. Međutim, povećana aktivnost koštane alkalne fosfataze zabilježena je u starijih kuja, što nije zapaženo u ostalih životinjskih vrsta niti zdravih ljudi. Također je uočena povećana razgradnja i smanjena izgradnja kosti u pasa veće tjelesne mase

Utjecaj selena u krvnom serumu na zdravstvene karakteristike vimena krava

The aim of this study was to determine the effect of selenium concentration in early lactation on mammary gland health and histological characteristics of the udder. The experiment included 30 high-yielding Holstein-Friesian cows. Selenium concentrations in blood and milk serum, and the average somatic cell count in the first and sixth months of lactation were analyzed. After exclusion of the experimental cows from the herd, histological characteristics of the udder were examined (the degree of leukocyte infiltration and the number of granulomas in the parenchyma). The mean selenium concentration in the blood serum was 0.62±0.11 mmol/L and that in the milk serum was 0.12±0.07 mmol/L. Optimal blood levels of selenium were found in 19 cows and suboptimal levels in 11 cows. A significant negative correlation was observed between blood and milk selenium concentrations and somatic cell count in early and mid lactation. There was no relationship between blood selenium concentration, milk selenium concentration and the amount of milk produced. Selenium-deficient cows had a significantly higher milk somatic cell count in early and mid lactation and significantly lower levels of selenium in milk. Upon histological analysis, 120 samples of individual quarters of the udder were grouped according to the degree of leukocyte infiltration and number of granulomas. Results showed that an increase in the degree of leukocyte infiltration and number of granulomas leads to an increase in the proportion of quarters from selenium-deficient cows and a decline in the percentage of quarters from cows exhibiting normal blood selenium concentrations. Selenium has a significant impact on udder health. Changes caused by selenium deficiency occur due to marked inflammation process in the mammary gland.Cilj rada bio je istražiti utjecaj koncentracije selena u ranoj laktaciji na zdravlje mliječne žlijezde i histološke karakteristike vimena krava. U istraživanje je bilo uključeno 30 visokomliječnih krava holštajn-frizijske pasmine. Analizirana je koncentracija selena u krvnom i mliječnom serumu, kao i prosječan broj somatskih stanica u prvom i šestom mjesecu laktacije. Nakon isključenja krava iz proizvodnje istraživana su histološka svojstva vimena krava (stupanj leukocitne infiltracije i broj granuloma u parenhimu). Prosječna koncentracija selena u krvnom serumu krava iznosila je 0,62±0,11 µmol/L, a u mliječnom 0,12±0,07µmol/L. U ogledu je utvrđeno 19 krava s optimalnom koncentracijom selena u krvi i 11 sa suboptimalnom koncentracijom. Utvrđena je značajna negativna korelacija koncentracije selena u krvi i mlijeku s brojem somatskih stanica u ranoj i srednjoj laktaciji. Nije utvrđena veza između koncentracije selena u krvi s koncentracijom selena u mlijeku i količinom proizvedenog mlijeka. Krave koje su deficitarne u selenu imaju značajno veći broj somatskih stanica u mlijeku u ranoj i srednjoj laktaciji i značajno nižu koncentraciju selena u mlijeku. Kada se nakon histološke analize 120 uzoraka pojedinačnih četvrti vimena grupiraju prema stupnju leukocitne infiltracije, odnosno prema broju granuloma, može se zaključiti da s porastom stupnja leukocitne infiltracije, odnosno broja granuloma u tkivu, raste udio onih četvrti koje potječu od krava deficitarnih u selenu, dok opada udio četvrti koje potječu od krava s normalnom koncentracijom selena u krvi. Selen značajno utječe na zdravlje vimena, a promjene koje izaziva deficit selena nastaju kao posljedica izražene inflamacije i reparacije u tkivu mliječne žlijezde

Uticaj organske i konvencionalne proizvodnje na sadržaj organske materije u černozemu

In this paper, in on-farm research, the effects of organic and conventional production systems on content of organic matter in Chernozem were compared. Preliminary results indicate that there were significant differences between some locations. Significantly lower organic matter content was measured on location Ljutovo (2,49%), compared to Čenej (3,17%) and Bačko Gradište (3,14%). In organic production systems which include manure application content of organic matter is equal or higher compared to conventional production systems. The lowest organic matter content was measured in crop rotations which include summer barley, regardless production systems or location.U radu su istraživanjima na farmi upoređeni uticaji organske i konvencionalne proizvodnje na sadržaj organske materije (OM) u černozemu. Preliminarni rezultati ukazuju da postoje značajne razlike između pojedinih lokaliteta. Znatno niži sadržaj organske materije izmeren je na lokalitetu Ljutovo (2,49%) u odnosu na lokalitete Čenej (3,17%) i Bačko Gradište (3,14%). U sistemima organske proizvodnje koji uključuju stajnjak, sadržaj organske materije u zemljištu je jednak ili visi u odnosu na sisteme konvencionalne proizvodnje. Najniži sadržaj organske materije izmeren je na plodoredima sa jarim ječmom, bez obzira na sistem proizvodnje i lokalitet