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

Phosphorus content in celeriac on the market of Zagreb

Celer je povrće koje se odlikuje visokom nutritivnom i zdravstvenom vrijednosti. Fosfor je u ljudskom organizmu od iznimne važnosti za žuč, bubrege i probavu. Dnevna potreba u ljudskoj ishrani za fosforom kod odraslog čovjeka iznosi 700 mg. Cilj ovog istraživanja bio je utvrditi količine fosfora u korijenu celera korjenaša prikupljenom na tržištu grada Zagreba. Uzorkovanje je provedeno 2016. na dvije tržnice, u dva trgovačka lanca i u dvije trgovine ekološkim proizvodima. Uzorci su sušeni na 105 °C do konstantne mase nakon čega su mljeveni i homogenizirani. Nakon razgradnje s HNO3 i HClO4 fosfor je određen spektrofotometrijski. Rezultati istraživanja pokazali su znatno variranje u količini fosfora u korijenu celera, u suhoj tvari od 0,50 do 0,93% P, a u svježoj tvari od 67 do 90 mg P/100 g svježe tvari, ovisno o prodajnom kanalu. Konzumacijom 100 g celera iz ovog istraživanja može se podmiriti od 9,8 do čak 12,8%. dnevnih potreba za fosforom.Celeriac is a vegetable of high nutrititve and health value. In human body, phosphorus is very imoprtant for bile, kidneys and digestion. The daily requirement in human nutrition for an adult is 700 mg of phosphorus. The aim of this research was to determine the phosphorus content of celeriac sampled on the market of Zagreb. Celeriac sampling was carried out in 2016 on 2 markets, in 2 market chains and in 2 stores with organic products. The samples were dried at 105 °C to a constant mass, after which were ground and homogenized. After digestion with HNO3 and HClO4 phosphorus was determined spectrophotometrically. The results of the research showed a significant variation in celeriac phosphorous content, in the dry weight from 0.50 to 0.93% P, and in the fresh weight from 67 to 90 mg P/100 g. By consuming 100 g of celeriac from this study 9.8 to 12.8% daily phosphorus needs can be satisfied

Methods for studying the localization of mitochondrial complexes III and IV by immunofluorescent and immunogold microscopy

The localization of proteins within a cell is very important for studying protein colocalization and subsequently understanding protein-protein interactions at the subcellular level. Using mitochondrial protein localization as a model, we established methods to study the localization of electron transport chain complexes (ETCCs), specifically complexes III and IV, in brown adipose tissue (BAT) and mitochondria. Immunofluorescent and immunogold techniques were applied to BAT paraffin sections and thin Araldite sections of mitochondria-enriched fractions, respectively. Microscopic analysis clearly showed mitochondrial localization of complexes III and IV, as well their colocalization. In addition, 10 and 20 nm gold particles were capable of identifying the localization of complexes within mitochondrial cristae. The methods described in this study may be a beneficial addition to currently utilized methods for accurately identifying the localization of ETCCs, their colocalization with other proteins and their distribution inside the cell and cellular compartments. Lastly, this method can also be used to study the molecular architecture of BAT mitochondria by analyzing fixed and postfixed thin plastic sections with electron microscopy (EM). [Projekat Ministarstva nauke Republike Srbije, br. 173054 i br.173055

Partial Elimination of Viruses from Traditional Potato Cultivar ‘Brinjak’ by Chemotherapy and Its Impact on Physiology and Yield Components

Viruses are responsible for more than 50% of annual potato tuber yield losses and cause great economic damage. The traditional Croatian potato cultivar ‘Brinjak’ is important for local growers because of its economically profitable production and as a gene pool for future breeding programs. However, the full genetic potential of the cultivar cannot be exploited due to virus infection. In this study, we attempted to eliminate potato virus M (PVM) and potato virus S (PVS) from potato cultivar ‘Brinjak’ and to evaluate the effects on physiological parameters and yield. Shoot apices were isolated from PVM + PVS-infected sprouts and cultivated for six weeks on MS medium with the addition of 50 or 100 mg L−1 ribavirin. The surviving shoot apices were micropropagated. The in vitro post-eradication period lasted 200 days. DAS-ELISA and RT-PCR were performed on R0 and R1 plants 90 days after acclimatization to determine the sanitary status of the plants. Chlorophyll fluorescence and multispectral imaging were performed on the R0 plants at the same time. The success of PVS elimination was 33% at both ribavirin concentrations. However, neither concentration was successful in eliminating PVM. Plants with mixed infection (PVM + PVS) had more severe disease symptoms compared to PVM-infected plants, affecting photochemistry and multispectral parameters and, consequently, yield. PVM + PVS plants had significantly lower number and weight of tubers per plant and lower average tuber weight than plants with single PVM infection in most of the generations studied. The results indicate a strong negative impact of PVS in mixed infections with PVM and show the importance of its elimination from potato plants

Partial Elimination of Viruses from Traditional Potato Cultivar ‘Brinjak’ by Chemotherapy and Its Impact on Physiology and Yield Components

Viruses are responsible for more than 50% of annual potato tuber yield losses and cause great economic damage. The traditional Croatian potato cultivar ‘Brinjak’ is important for local growers because of its economically profitable production and as a gene pool for future breeding programs. However, the full genetic potential of the cultivar cannot be exploited due to virus infection. In this study, we attempted to eliminate potato virus M (PVM) and potato virus S (PVS) from potato cultivar ‘Brinjak’ and to evaluate the effects on physiological parameters and yield. Shoot apices were isolated from PVM + PVS-infected sprouts and cultivated for six weeks on MS medium with the addition of 50 or 100 mg L−1 ribavirin. The surviving shoot apices were micropropagated. The in vitro post-eradication period lasted 200 days. DAS-ELISA and RT-PCR were performed on R0 and R1 plants 90 days after acclimatization to determine the sanitary status of the plants. Chlorophyll fluorescence and multispectral imaging were performed on the R0 plants at the same time. The success of PVS elimination was 33% at both ribavirin concentrations. However, neither concentration was successful in eliminating PVM. Plants with mixed infection (PVM + PVS) had more severe disease symptoms compared to PVM-infected plants, affecting photochemistry and multispectral parameters and, consequently, yield. PVM + PVS plants had significantly lower number and weight of tubers per plant and lower average tuber weight than plants with single PVM infection in most of the generations studied. The results indicate a strong negative impact of PVS in mixed infections with PVM and show the importance of its elimination from potato plants

Analiza promena na hrskavici i subhondralnoj kosti kiĉiĉnog zgloba radnog konja - radiološki i histološki pristup

Radni konji se u Srbiji koriste kao tovarne životinje koje nose/vuku tovar, često teži od njihove telesne mase. Postavili smo hipotezu da opterećenje koje trpe utiče na pojavu lezija na koštano-zglobnom sistemu. Cilj ovog rada je bio da se uporedno prikaže prisustvo i stepen oštećenja zglobne hrskavice (ZH) i subhondralne kosti (SHK), distalnih kondilusa treće metatarzalne kosti kod starijeg radnog konja pomoću makroskopskog pregleda hrskavice, radioloških i histoloških metoda. Na lokalnoj klanici uzorkovani su kičični zglobovi ekstremiteta radnog pastuva, starog 10 godina. Uz pomoć kompjuterizovane tomografije (CT) utvrđene su promene samo na zglobu prednje leve noge. Nakon disekcije označenog zgloba, izvršena je biopsija ZH i SHK metakarpalne kosti u predelu oštećenja, a bioptati su snimljeni mikro CT-om nakon čega su deklacifikovani i ukalupljeni u parafin, i analizirani su preseci. Makroskopski je bilo vidljivo da je lokalizacija oštećenja ZH bila simetrična na medijalnim i lateralnim epikondilusima matakarpalnih i metatarzalnih kostiju svih ekstremiteta. Linije habanja i palmarne artroze bile su slabo do umereno izražene. Na nativnim rendgenogramima zglobova nisu uočene patološke promene. Analizom snimaka načinjenih CT-om uočena je promena u subhondralnoj zoni distalne epifize treće metakarpalne kosti u vidu pukotine, a oko nje je primetna zona skleroze SHK. Na mikro CT snimku uzorka koštanog tkiva uzetog u predelu lateralnog epikondilusa metakarpane kosti, takođe je uočljiva zona subhondralne skleroze. Najmarkatnije histološke promene na ZH su bile: na površini ZH undulacije i fisure, koje se pružaju i do duboke zone, mnogobrojne izogene grupe sa većim brojem hondrocita u lakuni, fokalni gubitak ćelija i mestimičnio prisustvo hipertrofičnih hondrocita. U SHK su primećene: neznatne lezije koštanog matriksa u vidu svetlijeg prebojavanja, blago remodeliranje kosti koje se karakteriše njenim napredovanjem ka kalcifikovanoj hrskavici. Dugotrajno opterećenje utiče na pojavu oštećenja SHK koje se može uočiti uz pomoć CT-a i histološkom analizom, dok se stepen oštećenja ZH može evaluirati makroskopski i histološki.Zbornik kratkih sadržaj

Garlic Ecotypes Utilise Different Morphological, Physiological and Biochemical Mechanisms to Cope with Drought Stress

Drought negatively affects plants by altering morphological, physiological and metabolic processes and ultimately reducing yields. Garlic (Allium sativum L.), an important member of the Alliaceae family, is also sensitive to drought and maximizing the yield of garlic bulbs is largely dependent on water availability. The objective of this study was to determine the effects of drought stress on morphological and physiological characteristics, as well as on phenolic, sugar, inulin and free amino acid content and antioxidant activity in two Croatian garlic ecotypes, ‘Istarski crveni’ (IC) and Istarski bijeli (IB). Drought was induced by using polyethylene glycol 8000 (PEG) solution (−0.6 MPa) starting 21 days after clove planting and lasted for 20 days. Drought reduced plant height, number of leaves and plant weight, but increased root length in both ecotypes compared to the control treatment. Among the physiological parameters, significant differences were observed between the two ecotypes studied in the spectral characteristics of the leaves, namely reflection in red, green and blue, VAL, values of the vegetation indices related to the chlorophyll content (CHI, GI), and the anthocyanin content (ARI). Ecotype IC showed higher antioxidant activity in the control treatment due to higher total phenolic content (TPC), but under drought conditions higher DPPH radical scavenging activity was determined in ecotype IB and higher values of FRAP in IC. Sucrose and glucose generally decreased under drought, while inulin increased in IB but decreased in IC. Total free amino acid content increased under drought in both ecotypes. In conclusion, drought tolerance of IB might be associated with increased accumulation of inulin and higher levels of amino acids, especially those shown to contribute to drought resistance. In IC, drought tolerance is associated with an increase in some amino acid compounds and better root growth in depth, probably due to a more efficient translocation of sucrose to the underground part of the plant

Age-related Changes in the Articular Cartilage of the Stifle Joint in Non-working and Working German Shepherd Dogs

The aims of this study were to define age-related histological changes in the articular cartilage of the stifle joint in non-chondrodystrophic dogs and to determine whether physical activity has a positive impact on preservation of cartilage structure during ageing. Twenty-eight German shepherd dogs were included in the study. These dogs had no evidence of joint inflammation as defined by clinical assessment, radiology and synovial fluid analysis (specifically absence of synovial fluid serum amyloid A). The dogs were grouped as young working (n = 4), young non-working (n = 5), aged working (n = 13) and aged non-working (n = 6) animals. Gross changes in the stifle joints were recorded and biopsy samples of femoral and tibial articular cartilage were evaluated for thickness; chondrocyte number, density, surface area and morphology; isogenous group morphology; tidemark integrity; subchondral bone structure; presence of proteoglycans/glycosaminoglycans; and expression of type I, II and X collagens. The major age-related changes, not related to type of physical activity, included elevated chondrocyte density and thinning of tibial cartilage and increased chondrocyte surface area in the superficial and intermediate zone of the femoral cartilage. There was also expression of type X collagen in the femoral and tibial calcified and non-calcified cartilage; however, type X collagen was not detected in the superficial zone of old working dogs. Therefore, ageing, with or without physical activity, leads to slight cartilage degeneration, while physical activity modulates the synthesis of type X collagen in the superficial cartilage zone, partially preserving the structure of hyaline cartilage

A European map of groundwater pH and calcium

Water resources and associated ecosystems are becoming highly endangered due to ongoing global environmental changes. Spatial ecological modelling is a promising toolbox for understanding the past, present and future distribution and diversity patterns in groundwater-dependent ecosystems, such as fens, springs, streams, reed beds or wet grasslands. Still, the lack of detailed water chemistry maps prevents the use of reasonable models to be applied on continental and global scales. Being major determinants of biological composition and diversity of groundwater-dependent ecosystems, groundwater pH and calcium are of utmost importance. Here we developed an up-to-date European map of groundwater pH and Ca, based on 7577 measurements of near-surface groundwater pH and calcium distributed across Europe. In comparison to the existing European groundwater maps, we included several times more sites, especially in the regions rich in spring and fen habitats, and filled the apparent gaps in eastern and southeastern Europe. We used random forest models and regression kriging to create continuous maps of water pH and calcium at the continental scale, which is freely available also as a raster map (Hájek et al., 2020b; https://doi.org/10.5281/zenodo.4139912). Lithology had a higher importance than climate for both pH and calcium. The previously recognised latitudinal and altitudinal gradients were rediscovered with much refined regional patterns, as associated with bedrock variation. For ecological models of distribution and diversity of many terrestrial ecosystems, our new map based on field groundwater measurements is more suitable than maps of soil pH, which mirror not only bedrock chemistry but also vegetation-dependent soil processes.publishedVersio

A European map of groundwater pH and calcium

Water resources and associated ecosystems are becoming highly endangered due to ongoing global environmental changes. Spatial ecological modelling is a promising toolbox for understanding the past, present and future distribution and diversity patterns in groundwater-dependent ecosystems, such as fens, springs, streams, reed beds or wet grasslands. Still, the lack of detailed water chemistry maps prevents the use of reasonable models to be applied on continental and global scales. Being major determinants of biological composition and diversity of groundwater-dependent ecosystems, groundwater pH and calcium are of utmost importance. Here we developed an up-to-date European map of groundwater pH and Ca, based on 7577 measurements of near-surface groundwater pH and calcium distributed across Europe. In comparison to the existing European groundwater maps, we included several times more sites, especially in the regions rich in spring and fen habitats, and filled the apparent gaps in eastern and southeastern Europe. We used random forest models and regression kriging to create continuous maps of water pH and calcium at the continental scale, which is freely available also as a raster map (Hájek et al., 2020b; https://doi.org/10.5281/zenodo.4139912). Lithology had a higher importance than climate for both pH and calcium. The previously recognised latitudinal and altitudinal gradients were rediscovered with much refined regional patterns, as associated with bedrock variation. For ecological models of distribution and diversity of many terrestrial ecosystems, our new map based on field groundwater measurements is more suitable than maps of soil pH, which mirror not only bedrock chemistry but also vegetation-dependent soil processes