In an attempt to clarify the relationships between the digestive and metabolic effects of additional feeding of citrus pectin, the intestinal contents, tissues weights, nitrogen excretion and retention were studied in laboratory rats. Twenty-four growing male Wistar rats (98.8 g ± 5.3 g of body weight) were fed ad libitum for 20 days with balanced diets containing casein as the source of protein. In the experimental diet 96 g of wheat starch was replaced by 80 g of citrus pectin and 16 g of vegetable oil. Apparent digestibility and apparent protein biological value were calculated, the weights of digestive tissues and digestive organ content weights were also determined. All tissues of different parts of the digestive tract were heavier in the pectin fed group, and small intestine and caecum were statistically significant different as compared to control group. The contents of the small intestine and caecum were significant heavier in the pectin group. Pectin significantly lowered dry matter intake and growth rate and significantly increased faecal excretion of nitrogen and significantly decreased urinary nitrogen excretion. The consequence of alteration in the nitrogen excretion route was significantly lower apparent protein digestibility and apparent net protein utilisation, but apparent protein biological value was unchanged.

The article presents analysis of reasons for food dislikes among Slovenian primary schoolchildren aged 10 to 15, by gender, age and permanent residence. Altogether 628 primary school children from 16 randomly chosen schools in Slovenia filled the questionnaire. We found out that the most influential origins of food dislikes among schoolchildren were sensory characteristics, especially taste and smell. Among environmental factors were the most influential eating habits in families. Girls were more particular about their food then boys, except when it comes to vegetables. The study revealed that older schoolchildren disliked more organ meat than their younger colleagues. Differences among schoolchildren from western and eastern Slovenia were significant in dislikes toward meat from molluscs and amphibians. Schoolchildren from western Slovenia were less rejectable toward this kind of food.

Demanding transcriptomic studies in combination with challenging experiments in livestock animal species could be replaced by good in vitro models mimicking the function of ruminant mammary gland. The objective of our study was to establish epithelial cell line obtained from primary cell culture of lactating goat mammary gland. Mammary tissue from lactating Saanen goat (Capra hircus) was digested in collagenase and hyaluronidase solution and plated on plastic flasks. When growing on plastic, typical cobblestone morphology of epithelial cells and larger irregularly shaped cells, corresponding to myoepithelial cells were observed. When growth medium was supplemented with lactogenic hormones (insulin, hydrocortisone, and prolactin) and cells were cultured on plastic for extended period of time at high density, dome-like structures appeared as a result of cell to cell contact induced differentiation. Immunofluorescence staining using antibodies against smooth muscle a-actin, vimentin and various cytokeratins were used to distinguish between different cell types. Cell types of epithelial and myoepithelial cells were confirmed. Complete differentiation of cells was achieved when growing them on a commercial basal membrane matrix preparation which contains laminin, collagen IV, and various growth factors. Cells grown on basal membrane matrix in growth medium supplemented with lactogenic hormones differentiated morphologically and functionally. Spherical structures that resembled the alveoli of lactating mammary gland were observed. Reverse transcription PCR (RT-PCR) was performed on the total RNA extracted from the cultured cells in order to detect the potentially present milk protein mRNAs.

Cooperation between the ‘Panvita Group’ and the Biotechnical Faculty, University of Ljubljana, resulted in the construction of a pilot biogas reactor – a miniaturised version of the economical biogas reactor. The aim of construction was to support scientific research in the field of biogas generation, while, at the same time, optimising the processes conducted in economical reactors and testing the new substrates in the field of biogas generation. A 2500-litre reactor, containing a 500-litre gasholder, was built to this purpose. In the first operating period, biogas-generation potential of the raw and partially purified swine slurries was tested. The three repetitions allowed us to generate an average of 529 litres CH4 per 1 kilogram of organic dry matter.

A cryptic plasmid of approximately 3 kilobases named pTC18 was discovered in a ruminal Prevotella ruminicola TC18 strain and cloned into Escherichia coli. Based on pTC18, several shuttle vectors, containing Prevotella/Bacteroides tetQ selection marker and E. coli vector pUC19 inserted at two different positions in pTC18 were constructed. The shuttle vectors, protected with HaeIII methylase against the P. ruminicola 23 restriction were electroporated into P. ruminicola. Despite numerous attempts a tetracycline resistant recombinant strain 23 was not obtained. The possible causes for electroporation failure are discussed.

A survey of mountain pasture topsoil was undertaken first to set up field experiment in karst region on effects of applied P on minerals concentration in herbage. Content of SOM, C, N, CEC of soil and its base saturation are presented in the article. Great variability in depth, pH value and K level was found in soil. Low base saturation and high deficit of P was more common for all soil samples. Six different plant species presenting a great portion of herbage available for grazing and browsing at different occasions during grazing season were sampled and analysed for macro- and some microminerals. Concentration of P was very low in perennial grasses (1.1 g P kg-1 of DM). In leaves of common hazel (Corylus avellana L.) and common beech (Fagus silvatica L.) the concentration of P was identical as in white clover (Trifolium repens L.) and the level was high enough to cover animal needs when intake of herbage was sufficient. Leaves of woody plants were high in Mn concentration, but still below the levels that reduced growth rate in lambs. Application of P fertilizer had only small effect on increase of P in herbage, but large one on decrease on concentration of Ca in herbage. There was not clear effect of added P on concentration of Zn, Mn, Fe and Cu. Higher yield of DM induced with added fertilizer had not have any dilution effect on concentration of those minor elements in herbage.

Pedigree data from recorded flocks are of importance for the proper flock management as well as for the selection programmes and accurate performance prediction. In this study we examined if the use of eight molecular markers and computer analysis package ATLAS can be applied to verify known pedigrees of Jezersko-Solcava and Improved Jezersko-Solcava sheep breed from Slovenia. 90.9 % of pedigree data were in concordance with molecular genetic tests whereas in four pedigrees discordant parentage tests were obtained. In two cases, a different father or mother was assigned, whereas in the other two pedigrees both parents were discordant with molecular test results. In the second part of this study we aimed to determine the paternity for four lambs of Istrian Pramenka breed, in which a random mating scheme with 20 rams was used and hence the lambs had no father assigned. Using the same set of eight microsatellite markers, we were able to unequivocally determine paternity for 3 out of 4 lambs. In one case the analysis was not informative enough but with inclusion of 4 more microsatellite markers its sire could be determined. With the chosen set of microsatellite markers and data analysis programme ATLAS it is therefore possible to efficiently perform pedigree data validation as well as paternity prediction for lambs from flocks, where a large number of rams are used in a random mating system. Applying such molecular tests could help sheep breeders in flock management and improve efficiency of selection programmes.

We explored existing primer sequences targeting nitrous oxide reductase (nosZ) gene in order to explore their capability to recognize variant nosZ sequences. Published nosZ sequences longer than 380 AA residues were obtained from FunctionalGene Database /Repository (http://flyingcloud.cme.msu.edu/fungene/) and used for explorations with PrimerChart program. The numbers of sequences recovered using all possible forward and reverse primer combinations were determined and the stringency of primer site recognition was further varied by allowing 1, 2, or 3 primer mismatches to DNA binding site. We identified novel primer combinations resulting in satisfactory amplicon length (> 500 bp) and increased sequence recognition capabilities at comparable forward and reverse primer melting temperatures. Overall, this study indicates that current state of the art molecular methods can be and should frequently be further refined by the use of targeted bioinformatic approaches

This study aimed at evaluating the influence of strain and stocking density on welfare, haematological and serum biochemical indices of broilers in a 28-day trial. Two hundred and seven 4-week old birds each of Anak Titan and Arbor Acre genetic provenience were randomly allocated to three housing densities of 8.3, 11.1 and 14.3 birds/m2.These corresponded to 17, 22 and 30 birds per pen ( 2.01 × 1.00 m ) in a 2x3 factorial experiment. Each treatment group was replicated three times. The welfare parameters estimated were gait score, feather score, foot and hock burns, pecking, pushes, chases, fights and mortality. Blood samples were tested for packed cell volume (PCV), red blood cells (RBC), white blood cells (WBC), haemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC). Similarly, sera were utilized for the determination of total protein, albumin, globulin, glucose, cholesterol, and creatinine contents. There were no genotype-associated differences (P > 0.05) in the welfare indicators examined. However, placement density significantly (P0.05) in haematological profile. Strain and stocking density also exerted no influence (P > 0.05) on serum biochemical components. Strain × stocking density interaction effects were not observed in all the parameters. Consequently, the two strains could be reared at a density of 14.3 birds/m2 since density did not lead to a great degree of stress.

Only few plasmids and bacteriophages have been described to date in ruminal prevotella strains, therefore it appears plausible that the genetic exchange in these organisms must exploit other routes. Large conjugative transposons make possible the gene exchange process in bacteria from the genus Bacteroides, the phylogenetic relatives of ruminal prevotellas. The access to fully or partially finished genome sequences of Bacteroides and Prevotella representatives made possible the search for conserved regions within putative conjugative transposons. Multiple sequence alignment of known and putative conjugative transposon gene sequences of Bacteroides thetaiotaomicron, Prevotella intermedia, Bacteroides fragilis and Tanerella sp. was used to locate partially conserved regions within most preserved conjugative transposition genes, traG, and to construct appropriate degenerated oligonucleotide primers. These were used to amplify genome fragments from ruminal prevotella strains. Sequence analysis of the subcloned PCR products revealed the presence of a hypothetical gene in the genome of Prevotella bryantii B14, similar to the ORF BF2880 from B. fragilis YCH46, which is a part of a large conjugative transposon. Inverse PCRs were designed and performed to confirm the initial findings. A partial map of P. bryantii B14 putative conjugative transposon region was constructed, indicating an intergeneric horizontal gene transfer.

The time dependent formation of oxidative damage induced by polyunsaturated fat in the diet was investigated in an experiment with pigs as a model for humans. The role of vitamin E in the prevention of oxidative stress was also studied. Twenty-four growing pigs were penned individually and after an adaptation period divided into three groups. All groups received isocaloric daily rations composed of a basal diet isocalorically supplemented with: starch, linseed oil or linseed oil and vitamin E. Oxidative stress was evaluated by measuring the degree of lymphocyte and granulocyte nuclear DNA damage, concentration of malondialdehyde (MDA) in blood plasma, 24-hour urine MDA excretion rate and concentration of vitamin E isomers in the blood at the beginning, after 24 hours, after 6 days and at the end of the 22 day experimental period. The results confirmed that a high proportion of polyunsaturated fat in the diet increased lymphocyte and granulocyte DNA damage only after 6 days. The lymphocytes appear to be more sensitive to this type of oxidative stress than granulocytes. The MDA concentration in the blood and urinary MDA excretion after 24 hours of oxidative stress seem to be more accurate indicators than the rate of lymphocyte and especially granulocyte DNA damage. Vitamin E supplementation effectively protects the blood cells against increased DNA damage during the whole course of the experiment, but failed to reduce MDA formation significantly 24 hours and 6 days after the beginning of oxidative load. The study further suggests that supplementation of vitamin E is able to completely prevent DNA damage of both types of investigated blood cells at any time, but is only able to reduce the formation of lipid peroxidation products after prolonged treatment.

The ban on nutritive antibiotic use in Europe and the increased awareness of the consumers triggered a need for natural and safe feed additives to achieve better production results of farm animals. Plant extracts are used in animal nutrition as appetite and digestion stimulants, stimulants of physiological functions, for prevention and treatment of certain pathological conditions, as colorants and antioxidants. This article is a review of present literature data on the usage of plant extracts in poultry, pig and ruminant nutrition.