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IntroductionThe aim of our study was to measure fecal glucocorticoid metabolite (FGM) concentrations in captive and free-ranging male and female mountain gazelles (Gazella gazella) during their circannual cycle. In addition, FGM concentrations were used to track the intensity of the adrenocortical response in mountain gazelles during the same period.MethodsFecal samples were collected from the ground in the Hatay Mountain Gazelle Wildlife Development Area in the Hatay Province of Türkiye (36°32’ N, 36°32′ E) in each season of the year (December, April, July, September). The sex of the animals was determined by detecting the SRY gene of the Y chromosome in DNA isolated from the fecal samples. FGM was extracted from dried fecal samples with methanol, and its concentration was measured using a previously partially validated ELISA.Results and discussionThe results indicate that season is the most important factor explaining the variability in FGM concentrations in mountain gazelles. In animals of both sexes, the highest concentrations of FGM were observed in September. The values were significantly higher in the captive population, perhaps due to unpredictable stress. In July, FGM concentrations were low in both populations. As a result of the overall analysis across seasons, the comparison of FGM concentrations between captive and free-ranging animals revealed higher concentrations in captive animals only in September but not in other seasons, although higher concentrations have been previously reported for several wild captive species. Due to predation risk, the presence of offspring can be considered a critical point in the biological cycle for the welfare of free-ranging mountain gazelles, as suggested by the higher FGM concentrations in the free-ranging population in July. The high number of visitors could be a challenge for mountain gazelles in captivity, as indicated by higher FGM concentrations during September. Sex had no effect on the FGM concentrations of either population.

BackgroundProlonged hyperglycemia causes diabetes-related micro- and macrovascular complications, which combined represent a significant burden for individuals living with diabetes. The growing scope of evidence indicates that hyperglycemia affects the development of vascular complications through DNA methylation.MethodsA genome-wide differential DNA methylation analysis was performed on pooled peripheral blood DNA samples from individuals with type 1 diabetes (T1D) with direct DNA sequencing. Strict selection criteria were used to ensure two age- and sex-matched groups with no clinical signs of chronic complications according to persistent mean glycated hemoglobin (HbA1c) values over 5 years: HbA1c8% (N=10).ResultsBetween the two groups, 8385 differentially methylated CpG sites, annotated to 1802 genes, were identified. Genes annotated to hypomethylated CpG sites were enriched in 48 signaling pathways. Further analysis of key CpG sites revealed four specific regions, two of which were hypermethylated and two hypomethylated, associated with long non-coding RNA and processed pseudogenes.ConclusionsProlonged hyperglycemia in individuals with T1D, who have no clinical manifestation of diabetes-related complications, is associated with multiple differentially methylated CpG sites in crucial genes and pathways known to be linked to chronic complications in T1D.

The Lipizzan horse breed was established in the 16th century in Lipica and later spread throughout Central and Eastern Europe. In addition to six classical stallion lines, 17 classical mare family lines were established in the old Lipica stud. Additional 44 mare family lines, including local lines, were established in other Lipizzan studs. The founders of the classical Lipizzan mare family lines date back to the period between the 18th and 20th century and came from various breeds, including the Karst, Spanish, Italian, Kladruber and Arabian horses. The history of the Lipizzan mare family lines is well described and supported by the availability of 44 nucleotide sequences of the mtDNA control region. In this study, the mtDNA control region of 23 Serbian Lipizzans, representing the established Lippizan mare family lines, as well as the local mare family line Zora, have been sequenced. The objective of this study was to test whether the Zora mare family line could be distinguished from other Lipizzan mare family lines according to the mtDNA sequence. Sequences were aligned to the entire set of mtDNA control region sequences representing 44 Lipizzan mtDNA haplotypes. Based on the alignment, a median-joining network was constructed. The individuals in this study formed a cluster with the existing Lipizzan mare family lines. However, a total of ten individuals belonging to the local Zora mare family line formed a compact cluster close to the mtDNA haplotypes, representing the classical mare family lines Wera, Betalka, and Monteaura. Additional pedigree data are needed to clarify the origin of the mtDNA haplotype variant identified in this study.

Abstract Background Our aim was to determine whether child attachment to parents, parent attachment style, and morning cortisol levels were related to diabetes outcomes measured by average glycated hemoglobin (HbA1c), HbA1c variability over 4 years and time in range (TIR) in children with type 1 diabetes (T1D). Research design and methods 101 children with T1D and one of their parents were assessed at baseline for child attachment (Child Attachment Interview; CAI) and parent attachment (Relationship Structures Questionnaire; ECR-RS). Serum samples were collected for cortisol measurements before the interviews. HbA1c levels were measured during a 4-year follow-up period at regular 3-monthly visits, and data for TIR were exported from blood glucose measuring devices. Multivariate linear regression models were constructed to identify independent predictors of glycemic outcomes. Results More girls than boys exhibited secure attachment to their mothers. The results of the regression models showed that securely attached girls (CAI) had higher average HbA1c than did insecurely attached girls (B = −0.64, p = 0.03). In boys, the more insecure the parent's attachment style, the worse the child's glycemic outcome: the higher the average Hb1Ac (B = 0.51, p = 0.005), the higher the HbA1c variability (B = 0.017, p = 0.011), and the lower the TIR (B = −8.543, p = 0.002). Conclusions Attachment in close relationships is associated with glycemic outcomes in children with T1D, and we observed significant differences between sexes. A sex- and attachment-specific approach is recommended when treating children with less favorable glycemic outcomes. Special attention and tailored support should be offered to securely attached girls in transferring responsibility for diabetes care and at least to male children of insecurely attached parents to prevent suboptimal glycemic control. Further studies in larger samples and more daily cortisol measurements may help us better understand the links between stress response, attachment and T1D.

The domestic cat (Felis silvestris catus) is a valued companion animal throughout the world. Over 60 different cat breeds are accepted for competition by the cat fancy registries in different countries. Genetic markers, including short tandem repeats and SNPs, are available to evaluate and manage levels of inbreeding and genetic diversity, population and breed structure relationships, and individual identification for forensic and registration purposes. The International Society of Animal Genetics (ISAG) hosts the Applied Genetics in Companion Animals Workshop, which supports the standardization of genetic marker panels and genotyping for the identification of cats via comparison testing. SNP panels have been in development for many species, including the domestic cat. An ISAG approved core panel of SNPs for use in cat identification and parentage analyses is presented. SNPs (n = 121) were evaluated by different university-based and commercial laboratories using 20 DNA samples as part of the ISAG comparison testing procedures. Different SNP genotyping technologies were examined, including DNA arrays, genotyping-by-sequencing and mass spectroscopy, to select a robust and efficient panel of 101 SNPs as the ISAG core panel for cats. The SNPs are distributed across all chromosomes including two on the X chromosome and an XY pseudo-autosomal sexing marker (zinc-finger XY; ZFXY). A population study demonstrated that the markers have an average polymorphic information content of 0.354 and a power of exclusion greater than 0.9999. The SNP panel should keep testing affordable while also allowing for the development of additional panels to monitor health, phenotypic traits, hybrid cats and highly inbred cats.

Adipogeneza je kompleksen proces, na katerega vpliva veliko število genetskih in okoljskih faktorjev. Ker je posledica prekomernega nalaganja maščob debelost, pogosto pa tudi pojav diabetesa tipa 2, postaja proučevanje molekulskih osnov adipogeneze vse bolj pomembno tudi z vidika zdravja človeka. V živalski proizvodnji proces tvorbe maščobnega tkiva pomembno vpliva na kakovost prirasta, preko porabe krme pa na ekonomičnost proizvodnje. Poleg leptina in leptinskega receptorja, ki odločilno uravnavata nalaganje maščob, je bil v zadnjem času odkrit tudi mehanizem uravnavanja tvorbe maščobnega tkiva, ki vključuje adaptivno termogenezo in biosintezo mitohondrijev. Pomembna regulatorja teh procesov sta PPARg in njegov koaktivator PGC-1a. V pričujočem prispevku opisujemo mehanizem uravnavanja adipogeneze in vlogo PPARg in PGC-1a v njem. Prikazana je tudi medvrstna primerjava aminokislinskih zaporedij PGC-1a in možnosti za razvoj farmacevtskih učinkovin, ki bi vplivale na izražanje PGC-1a.

Relative energy deficiency syndrome develops in children and adolescents as a result of a negative energy balance due to insufficient energy intake during intense physical exercise regimens. This energy imbalance leads to impaired physiological function of multiple organ systems. Moreover, in children and adolescents, it presents a risk for the development of long-term consequences in terms of negative effects on growth, pubertal development and bone health. Early recognition and management are therefore crucial in this age group. Due to poor awareness among young athletes, their parents, coaches, medical staff, and a non-specific clinical presentation the illness is often overlooked.

Sindrom relativnega energijskega primanjkljaja se razvije pri otrocih in mladostnikih športnikih ob negativni energijski bilanci, ki je posledica nezadostnega vnosa energije glede na velike potrebe ob intenzivni telesni vadbi. Energijsko neravnovesje privede do motenj v delovanju več organskih sistemov. Pri otrocih in mladostnikih so posledice lahko dolgoročne in se kažejo kot neugoden vpliv na rast, pubertetni razvoj in zdravje kosti. Zgodnji prepoznava in obravnava sta tako v tej starostni skupini ključni. Zaradi pomanjkljive ozaveščenosti med mladimi športniki, njihovimi starši, trenerji in zdravstvenim osebjem ter nespecifične klinične slike bolezen pogosto spregledamo.

Izhodišča: Test QuantiFERON TB (QFT TB) je prvenstveno namenjen za odkrivanje latentne (LTBI) okužbe z bacili tuberkuloze (TB). Članek obsega retrospektivno analizo rezultatov testa QFT TB v Sloveniji med letoma 2008 in 2019, opozarja na vplive predanalitskih dejavnikov na rezultate testa ter govori o razlogih za nastanek nejasnih rezultatov in o občutljivosti testa pri bolnikih z aktivno obliko TB. Metode: V obdobju 2008–2019 smo v Laboratoriju za mikobakterije Klinike Golnik s testom QFT TB testirali 29.352 vzorcev krvi bolnikov iz različnih zdravstvenih ustanov v Sloveniji. Na rezultatih testa QFT TB smo izvedli retrospektivno analizo. Rezultati: Delež pozitivnih rezultatov testa QFT TB se z leti postopno znižuje. V letu 2008 je znašal 20,8 %, v letu 2019 pa 10,7 %, kar pomeni, da je v zadnjih dvanajstih letih upadel za 48,6 %. Največjo težavo predstavljajo nejasni rezultati (letno povprečno 4,3 %), saj so za zdravnika nepovedni. Občutljivost testa QFT TB pri bolnikih z aktivno obliko TB je 82,9 %, kar je primerljivo s podatki iz tujine. Zaključek: Pomembno je, da pri odvzemu in pošiljanju krvi dosledno sledimo navodilom, saj je test QFT TB zelo občutljiv na predanalitske dejavnike in klinično stanje preiskovancev. Delež pozitivnih rezultatov testa QFT TB z leti pospešeno upada, kar je povezano z napori za omejitev širjenja TB in upadom primerov aktivne TB v naši državi.

Animal welfare is a multidimensional concept that includes several physical and psychological parameters of the animal. The aim of this study was to assess animal welfare during transportation and in Slovenian beef, pork, and poultry slaughterhouses. A questionnaire was used for this study. Several parameters of animal welfare were rated on a 5-point scale, such as health status, animal behavior, lairage or transport vehicle conditions, and driver regulation compliance. The scale was also used for the second part of the study. This consisted of two studies: (1) self-assessment by slaughterhouse employees and livestock transport drivers and (2) animal welfare observational assessment performed by two veterinarians. The results were compared with each other. Ten large slaughterhouses and nine livestock drivers took part in the survey. The results showed that slaughterhouse employees knew more about animal welfare than livestock truck drivers, but both groups were not sufficiently familiar with animal welfare laws and regulations. This study found that the experts’ assessments were generally lower than the self-assessments of employees and livestock drivers. Based on the research findings, it can be concluded that there is a need to improve the awareness and knowledge of slaughterhouse employees and livestock drivers regarding animal handling and animal welfare regulations.

Marek’s disease (MD), caused by Mardivirus gallidalpha 2 (GaAHV-2), also known as MD virus (MDV), is a lymphoproliferative disease that primarily affects chickens. Recently, MDV has been detected in lymphomatous tumors in turkeys in various countries. Between 2021 and 2023, three cases ranging from no to severe clinical disorders (depression, lameness, and increased mortality) occurred in commercial turkey flocks in Slovenia. In all cases, MDV was detected by PCR in DNA samples extracted from organs developing tumor infiltrations. Sequencing and phylogenetic analysis of the meq gene revealed that the GaAHV-2 detected has molecular features of a very virulent pathotype and genetic similarity with GaAHV-2 detected in chickens in Tunisia. This is the first report of MDV in commercial turkeys in Slovenia.

ABSTRACT: Objectives: Next-generation sequencing (NGS) can provide a comprehensive analysis of the genetic alterations most commonly linked with pyrazinamide (PZA) resistance. However, there are no studies reporting the molecular background of PZA resistance in Mycobacterium tuberculosis (TB) isolates from the Balkan Peninsula. We aimed to examine the feasibility of full-length analysis of a gene linked with PZA resistance (pncA) using Ion Torrent technology compared with phenotypic BACTEC MGIT 960 drug susceptibility testing (DST) in clinical TB isolates from two countries of the Balkan Peninsula. Methods: Between 1996 and 2017, we retrospectively selected 61 TB isolates. To identify gene variants related to drug resistance in genomic DNA extracted from TB isolates, AmpliSeq libraries were generated automatically using an AmpliSeq™ Kit for Chef DL8 and Ion AmpliSeq TB Research Panel. Results: Of the 61 TB isolates, 56 were phenotypically resistant to any antibiotic. Among them, 38/56 isolates (67.9%) were phenotypically resistant to pyrazinamide, and pncA mutations were detected in 34/38 cases (89.5%). A mutation in the pncA promoter region was the most prevalent genetic alteration, detected in eight TB isolates. Comparison of NGS data with conventional BACTEC MGIT 960 DST revealed very strong agreement (91.8%) between the two methods in identifying PZA resistance, with high sensitivity (89.5%) and specificity (95.7%) for NGS. Conclusion: Detection of PZA resistance using NGS appears to be a valuable tool for surveillance of TB drug resistance in the Balkan Peninsula, with great potential to provide useful information at least 1 weak earlier than is possible with phenotypic DST.

Abstract The contribution of domestic cattle in human societies is enormous, making cattle, along with other essential benefits, the economically most important domestic animal in the world today. To expand existing knowledge on cattle domestication and mitogenome diversity, we performed a comprehensive complete mitogenome analysis of the species (802 sequences, 114 breeds). A large sample was collected in South‐east Europe, an important agricultural gateway to Europe during Neolithization and a region rich in cattle biodiversity. We found 1725 polymorphic sites (810 singletons, 853 parsimony‐informative sites and 57 indels), 701 unique haplotypes, a haplotype diversity of 0.9995 and a nucleotide diversity of 0.0015. In addition to the dominant T3 and several rare haplogroups (Q, T5, T4, T2 and T1), we have identified maternal line in Austrian Murbodner cattle that possess surviving aurochs’ mitochondria haplotype P1 that diverged prior to the Neolithization process. This is convincing evidence for rare female‐mediated adaptive introgression of wild aurochs into domesticated cattle in Europe. We revalidated the existing haplogroup classification and provided Bayesian phylogenetic inference with a more precise estimated divergence time than previously available. Occasionally, classification based on partial mitogenomes was not reliable; for example, some individuals with haplogroups P and T5 were not recognized based on D‐loop information. Bayesian skyline plot estimates (median) show that the earliest population growth began before domestication in cattle with haplogroup T2, followed by Q (~10.0–9.5 kyBP), whereas cattle with T3 (~7.5 kyBP) and T1 (~3.0–2.5 kyBP) expanded later. Overall, our results support the existence of interactions between aurochs and cattle during domestication and dispersal of cattle in the past, contribute to the conservation of maternal cattle diversity and enable functional analyses of the surviving aurochs P1 mitogenome.

Abstract Background The importance of local breeds as genetic reservoirs of valuable genetic variation is well established. Pig breeding in Central and South-Eastern Europe has a long tradition that led to the formation of several local pig breeds. In the present study, genetic diversity parameters were analysed in six autochthonous pig breeds from Slovenia, Croatia and Serbia (Banija spotted, Black Slavonian, Turopolje pig, Swallow-bellied Mangalitsa, Moravka and Krskopolje pig). Animals from each of these breeds were genotyped using microsatellites and single nucleotide polymorphisms (SNPs). The results obtained with these two marker systems and those based on pedigree data were compared. In addition, we estimated inbreeding levels based on the distribution of runs of homozygosity (ROH) and identified genomic regions under selection pressure using ROH islands and the integrated haplotype score (iHS). Results The lowest heterozygosity values calculated from microsatellite and SNP data were observed in the Turopolje pig. The observed heterozygosity was higher than the expected heterozygosity in the Black Slavonian, Moravka and Turopolje pig. Both types of markers allowed us to distinguish clusters of individuals belonging to each breed. The analysis of admixture between breeds revealed potential gene flow between the Mangalitsa and Moravka, and between the Mangalitsa and Black Slavonian, but no introgression events were detected in the Banija spotted and Turopolje pig. The distribution of ROH across the genome was not uniform. Analysis of the ROH islands identified genomic regions with an extremely high frequency of shared ROH within the Swallow-bellied Mangalitsa, which harboured genes associated with cholesterol biosynthesis, fatty acid metabolism and daily weight gain. The iHS approach to detect signatures of selection revealed candidate regions containing genes with potential roles in reproduction traits and disease resistance. Conclusions Based on the estimation of population parameters obtained from three data sets, we showed the existence of relationships among the six pig breeds analysed here. Analysis of the distribution of ROH allowed us to estimate the level of inbreeding and the extent of homozygous regions in these breeds. The iHS analysis revealed genomic regions potentially associated with phenotypic traits and allowed the detection of genomic regions under selection pressure.

Hipokalcemija je pogosta motnja v presnovi kalcija in fosfata pri otrocih, zlasti pri novorojenčkih. Pogosto ne povzroča simptomov. Simptomi in znaki so odvisni od otrokove starosti. Na primarni ravni zdravstvene obravnave zdravnik diagnozo hipokalcemija potrdi z laboratorijsko analizo krvi in otroka klinično stabilizira. Nadaljnjo laboratorijsko in slikovno diagnostično obravnavo opravimo na sekundarni oz. največkrat na terciarni ravni. V prispevku predstavljamo primer deklice s klinično izraženo hipokalcemijo zaradi primarnega hipoparatiroidizma ob avtoimunskem poliglandularnem sindromu tipa 1.

Hypocalcaemia is a common disorder of calcium and phosphate metabolism in children, especially in newborns. It is often asymptomatic; clinical signs and symptoms are dependent on the age of the child. The primary care physician can confirm hypocalcaemia in the child with a suspicious clinical picture and clinically stabilise the child. Further laboratory and imaging diagnostics should be performed at the secondary and most often the tertiary level of medical care. In the article, the case of a girl with clinically manifested hypocalcaemia, caused by primary hypoparathyroidism as a consequence of autoimmune polyglandular syndrome type 1, is presented.

The usefulness of blood collection using venipuncture versus kissing bugs or medicinal leeches and the collection of saliva, faeces, hair, urine, and tears for measuring “immunoreactive” C (iC) concentration in Alpine ibexes was verified using commercial enzyme immunoassays. The mean value of serum C was highest in serum collected using venipuncture and lowest in serums collected using kissing bugs. Statistically significant differences were observed between venipuncture and kissing bugs and between leeches and kissing bugs. However, no statistically significant difference was found in C concentrations between samples collected with venipuncture and those collected with leeches. The highest mean value of C concentration was measured in serum (all three methods), followed by that in hair and faeces, and the lowest mean value was found in saliva. Statistically significant differences were found between saliva and faeces samples and between saliva and hair samples. The difference between the concentrations for faeces and hair was not statistically significant. A significant difference in C concentration between males and females was found in saliva. A significant difference in C concentration among different ages was measured in serum obtained using venipuncture in all three groups and in faeces between the groups older than ten years and younger than 10 months. Highly significant differences in C concentrations were also found between hair sampled in summer and hair sampled in autumn. Collecting tear and urine samples is a laborious procedure and is therefore less acceptable for C determination. Due to the small number of samples, statistical values are not given for these two matrices.

Animal welfare is a multiparameteral concept that encompasses the physical and mental health of animals and includes various aspects such as physical wellbeing, absence of hunger and thirst, and ability to express motivated behavior, to which farmers usually attach different importance. The objectives of this study were to evaluate animal welfare on Slovenian commercial pig farms, to determine whether farmers' perceived importance of animal welfare differ from actual animal welfare on farms and to determine, if farmer's age, gender, their level of education and participation in vocational training have an influence. For that purpose, we created an Animal Welfare Protocol/Questionnaire for Pig Farms (AWQ/P-P) that assessed several parameters of animal welfare: (1) general status, (2) animal behavior, (3) health status, (4) living conditions, and (5) environmental conditions. Each parameter included at least five observation points and was scored on a 5-point scale. The same observation points were used to measure farmers' perceived importance of animal welfare and for observational assessment. Consequently, we were able to compare both statistically. Farmers from 14 (N = 14) large Slovenian pig farms participated in the study. Results show that farmers rate all parameters of animal welfare very highly. For them, animal health status is the most important, and environmental conditions are the least important factors for animal welfare. Observational inspections yielded significantly lower scores for animal welfare conditions than those obtained from farmer ratings. The highest correlations between farmers' perceptions and observational inspections were found for the parameters of animal behavior and environmental conditions. The results of this study also suggest that vocational training is a significant variable in increasing levels of pig welfare. Age, gender, and education level are not significant variables, however, farms led by older male farmers with lower level of education but involved in vocational training from different sources had slightly better welfare on the farm. This should be further investigated before making conclusions, due to our small sample size. The significance of the study is to identify deficiencies in pig welfare as perceived by farmers and consequently improve pig welfare.

Abstract Background Local breeds retained unique genetic variability important for adaptive potential especially in light of challenges related to climate change. Our first objective was to perform, for the first time, a genome-wide diversity characterization using Illumina GoatSNP50 BeadChip of autochthonous Drežnica goat breed from Slovenia, and five and one local breeds from neighboring Austria and Italy, respectively. For optimal conservation and breeding programs of endangered local breeds, it is important to detect past admixture events and strive for preservation of purebred representatives of each breed with low or without admixture. In the second objective, we hence investigated the effect of inclusion or exclusion of outliers from datasets on genetic diversity and population structure parameters. Results Distinct genetic origin of the Drežnica goat was demonstrated as having closest nodes to Austrian and Italian breeds. A phylogenetic study of these breeds with other goat breeds having SNP data available in the DRYAD repository positioned them in the alpine, European and global context. Swiss breeds clustered with cosmopolitan alpine breeds and were closer to French and Spanish breeds. On the other hand, the Drežnica goat, Austrian and Italian breeds were closer to Turkish breeds. Datasets where outliers were excluded affected estimates of genetic diversity parameters within the breed and increased the pairwise genetic distances between most of the breeds. Alpine breeds, including Drežnica, Austrian and Italian goats analyzed here, still exhibit relatively high levels of genetic variability, homogeneous genetic structure and strong geographical partitioning. Conclusions Genetic diversity analyses revealed that the Slovenian Drežnica goat has a distinct genetic identity and is closely related to the neighboring Austrian and Italian alpine breeds. These results expand our knowledge on phylogeny of goat breeds from easternmost part of the European Alps. The here employed outlier test and datasets optimization approaches provided an objective and statistically powerful tool for removal of admixed outliers. Importance of this test in selecting the representatives of each breed is warranted to obtain more objective diversity parameters and phylogenetic analysis. Such parameters are often the basis of breeding and management programs and are therefore important for preserving genetic variability and uniqueness of local rare breeds.

Background: Haemophilic arthropathy is often associated with a loss of range of motion. Total knee arthroplasty is an effective treatment option for patients with end-stage haemophilic arthropathy of the knee. However, even after arthroplasty, the range of motion sometimes remains insufficient. Objective: To evaluate static progressive stretch as a treatment method for haemophilic patients with decreased range of motion after total knee arthroplasty. Methods: Static progressive stretch was used to improve range of motion in patients with a postoperative extension lag of more than 10° and flexion of less than 80°. A total of 7 knees were treated for a mean of 21.7 weeks. Results: Statistically significant increases in range of motion and in Knee Society Score were observed when comparing pre-treatment and post-treatment values. Conclusions: Static progressive stretch using an orthotic device could be a successful adjuvant method for treating joint stiffness in patients with haemophilia after total knee arthroplasty. LAY ABSTRACT Haemophilic arthropathy often results in loss of range of motion. Total knee arthroplasty is an effective treatment option for patients with end-stage haemophilic arthropathy of the knee. However, despite arthroplasty, the range of motion is sometimes insufficient. Static progressive stretch using special knee orthoses is an adjunct technique to traditional physiotherapy, intending to restore functional range of motion and decrease knee stiffness and pain. The aim of this study was to evaluate static progressive stretch as a treatment method for hemophilia patients with decreased range of motion after total knee arthroplasty.

Encephalitozoon cuniculi is a microsporidial parasite that primarily infects domestic rabbits (Oryctolagus cuniculus). It is the causative agent of encephalitozoonosis, a disease with an internationally recognized seroprevalence among rabbits. This study determines the presence, clinical manifestation, and serological status of encephalitozoonosis in pet rabbits in Slovenia using various diagnostic procedures. From 2017 to 2021, 224 pet rabbit sera were collected and tested for encephalitozoonosis with the indirect immunofluorescence assay. Immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies against E. cuniculi were confirmed in 160 (65.6%) cases. Most seropositive rabbits suffered from neurological clinical signs or signs of gastrointestinal disorders such as recurrent hypomotilities, chronic weight loss, cachexia, or anorexia, and fewer showed clinical signs related to the urinary system or phacoclastic uveitis. A quarter of the positively tested rabbits presented without clinical signs. Hematological and biochemical blood analysis confirmed that seropositive animals had elevated globulin and deviated albumin levels in comparison to the normal reference values of non-infected animals. Furthermore, rabbits with neurological clinical signs showed statistically significant higher levels of globulins and total protein. Sixty-eight whole-body radiographs and thirty-two abdominal ultrasound reports were analyzed, looking for changes in the shape or size of the urinary bladder, presence of urinary sludge or uroliths, and any abnormalities related to the kidneys (shape, size, or nephrolites). The results suggest that neurological defects in the urinary bladder caused by E. cuniculi lead to a distended urinary bladder and consequently dysuria, incontinence, urine scalding, and sludgy urine.

Type 1 diabetes (T1D) is one of the most common chronic diseases of the endocrine system, associated with several life-threatening comorbidities. While the etiopathogenesis of T1D remains elusive, a combination of genetic susceptibility and environmental factors, such as microbial infections, are thought to be involved in the development of the disease. The prime model for studying the genetic component of T1D predisposition encompasses polymorphisms within the HLA (human leukocyte antigen) region responsible for the specificity of antigen presentation to lymphocytes. Apart from polymorphisms, genomic reorganization caused by repeat elements and endogenous viral elements (EVEs) might be involved in T1D predisposition. Such elements are human endogenous retroviruses (HERVs) and non-long terminal repeat (non-LTR) retrotransposons, including long and short interspersed nuclear elements (LINEs and SINEs). In line with their parasitic origin and selfish behaviour, retrotransposon-imposed gene regulation is a major source of genetic variation and instability in the human genome, and may represent the missing link between genetic susceptibility and environmental factors long thought to contribute to T1D onset. Autoreactive immune cell subtypes with differentially expressed retrotransposons can be identified with single-cell transcriptomics, and personalized assembled genomes can be constructed, which can then serve as a reference for predicting retrotransposon integration/restriction sites. Here we review what is known to date about retrotransposons, we discuss the involvement of viruses and retrotransposons in T1D predisposition, and finally we consider challenges in retrotransposons analysis methods.

The aim of this study was to determine the percentage of hatched and fertilized eggs in female Hermann’s tortoises before and after the removal of males after breeding.

Ščitnica je ektopična, če ščitnično tkivo leži na netipičnem anatomskem mestu. Je posledica motnje v njenem embri- onalnem razvoju. Najpogosteje se nahaja na korenu jezi- ka, najdemo pa jo tudi drugje v poteku spusta ščitnice med embriogenezo in celo na oddaljenih mestih. Večinoma je enojna, redkeje pa opisujejo tudi primere dvojne ali celo trojne ektopije. Osebe z ektopično ščitnico navadno nimajo nikakršnih simptomov. Sicer so simptomi odvisni od lege in velikosti ektopičnega tkiva ter od njegove endokrine disfunkcije. Najpomembnejša preiskava za opredelitev mesta ektopične ščitnice je scintigrafija. »eprav pri brezsimptomnih bolnikih zdravljenje ni potrebno, moramo poskrbeti za redno slede- nje in ustrezno ukrepanje ob pojavu zapletov. Podobno kot v ortotropni ščitnici lahko tudi v ektopični ščitnici pride do bolezenskih sprememb. V prispevku predstavljamo redek primer dvojne ščitnične ektopije s subkliničnim hipotiroidizmom pri dve in pol letni deklici in podajamo pregled primerov ektopične ščitnice, ki smo jih v 5-letnem obdobju diagnosticirali na Pediatrični kliniki Ljubljana.

Ectopic thyroid gland is a rare clinical entity characterized by thyroid tissue in a location other than its normal anatom- ical position. It is a developmental abnormality. The most frequent location of ectopic thyroid is at the base of the tongue, although it can be found elsewhere along the path of thyroid embryogenic descent and even at distant body locations. Single ectopy is the most common, but dual and triple ectopy has also been described. Most people with ectopic thyroid are asymptomatic. If symptoms occur, they are related to the location and size of the ectopic tissue as well as its endocrine dysfunction. The most important diagnostic procedure for detection of ectopic thyroid tissue is scintigraphy. For patients with- out symptoms, no treatment is necessary, but they need regular follow-up and action if symptoms appear. Patho- logical changes can occur in the ectopic as in the ortho- topic gland. We present a rare case of dual ectopy with subclinical hypo- thyroidism in a two-and-a-half-year-old girl and a review of cases of ectopic thyroid diagnosed in a 5-year period at the University Children’s Hospital Ljubljana.

Drug resistant tuberculosis (TB), especially multidrug (MDR) and extensively drug-resistant (XDR) TB, is still a serious problem in global TB control. Slovenia and North Macedonia are low-incidence countries with TB incidence rates of 5.4 and 10.4 in 2017, respectively. In both countries, the percentage of drug resistant TB is very low with sporadic cases of MDR-TB. However, global burden of drug-resistant TB continues to increase imposing huge impact on public health systems and strongly stimulating the detection of gene variants related with drug resistance in TB. Next-generation sequencing (NGS) can provide comprehensive analysis of gene variants linked to drug resistance in Mycobacterium tuberculosis. Therefore, the aim of our study was to examine the feasibility of a full-length gene analysis for the drug resistance related genes (inhA, katG, rpoB, embB) using Ion Torrent technology and to compare the NGS results with those obtained from conventional phenotypic drug susceptibility testing (DST) in TB isolates. Between 1996 and 2017, we retrospectively selected 56 TB strains from our National mycobacterial culture collection. Of those, 33 TB isolates from Slovenian patients were isolated from various clinical samples and subjected to phenotypic DST testing in Laboratory for Mycobacteria (University Clinic Golnik, Slovenia). The remaining 23 TB isolates were isolated from Macedonian patients and sent to our laboratory for assistance in phenotypic DST testing. TB strains included were either mono-, poly- or multidrug resistant. For control purposes, we also randomly selected five TB strains susceptible to first-line anti-TB drugs. High concordance between genetic (Ion Torrent technology) and standard phenotypic DST testing for isoniazid, rifampicin and ethambutol was observed, with percent of agreement of 77%, 93.4% and 93.3%, sensitivities of 68.2%, 100% and 100%, and specificities of 100%, 80% and 88.2%, respectively. In conclusion, the genotypic DST using Ion Torrent semiconductor NGS successfully predicted drug resistance with significant shortening of time needed to obtain the resistance profiles from several weeks to just a few days.

Natančna opredelitev antigena (Ag) RhD (D) je pomembna pri nosečnicah, prejemnikih transfuzij in krvodajalcih. Glede na tip D se pri nosečnicah odločamo o pred- in poporodni zaščiti z imunoglobulinom anti-D (Ig anti-D). Glede na določitev Ag D lahko posameznike razvrstimo med D-pozitivne (D-poz) ali D-negativne (D-neg), obstajajo pa tudi številne druge različice Ag D (variante D, D-var). Variantni aleli gena RHD (D) lahko nosijo zapis za različne oblike proteina D, razdelimo jih lahko na šibke, parcialne in Del. Ag D določamo s serološkimi metodami, v primeru nejasnosti si pomagamo z molekularnobiološkimi metodami za določitev gena D. Dokončno opredelitev D-var podamo na podlagi molekularnobioloških metod. Nosečnicam smo iz vzorcev periferne venske krvi določili Ag D na ploščici (reagent Seraclone anti-D) in na gelu (testni sistem z gelskimi karticami DiaClon ABO/D + Reverse Grouping, monoclonal antibodies, Bio-Rad, Nemčija, s katerimi ne zaznamo krvne skupine D kategorija VI). 49 nosečnicam smo opravili razširjeno serološko testiranje KS D (komercialni set ID-Partial RhD Typing Set, Bio-Rad, Nemčija). Nadaljevali smo z molekularnobiološkimi metodami: osamitev DNA (uporaba aparata BioRobot EZ1 in komercialnega set EZ1 DNA Blood 350 μl Kit, Qiagen, Nemčija), določitev genotipa D z metodo PCR-SSP (komercialni seti RBC-Ready Gene CDE, RBC-Ready Gene D weak in RBC-Ready Gene D AddOn, Inno-Train; Nemčija), pomnoževanje (aparat Veriti, Apllied BioSystems, ZDA), ločevanje produktov (elektroforezni system, BioRad, ZDA) na agaroznem gelu (Sigma, Nemčija). Ugotovili smo, da so šibke oblike D predstavljale 41 primerov (83,7 %), parcialne oblike D pa 8 primerov (16,3 %) vseh D-var. Najpogostejša oblika D-var med nosečnicami je bila šibki D tip 1 s 17 primeri (34,7 %), sledil je šibki D tip 3 s 13 primeri (26,5 %) in šibki D tip 2 z 10 primeri (20,4 %). Najpogostejša parcialna oblika D je bila D kategorija VII s 5 primeri (10,2 %). Nosečnice in prejemnike transfuzij, ki so nosilci šibkih D tipa 1, 2 ali 3, lahko varno obravnavamo kot D-poz, nosilce vseh ostalih D-var pa kot D-neg. Krvodajalce z D-var obravnavamo kot D-poz. S tem algoritmom lahko preprečimo približno 172 nepotrebnih aplikacij Ig anti-D letno in ohranjamo tudi primerne zaloge D-neg eritrocitnih komponent krvi.

Introduction: A traumatic event is an extremely threatening and frightening experience in an individual's life. Children who are exposed to traumatic events are twice as likely to develop a mental disorder. Screening can provide insight into the traumatic experience of children, identifying those eligible for further evaluation, and support. With this aim, we evaluated the psychometric properties of the Lifetime Incidence of Traumatic Events questionnaire (LITE) in Slovene by calculating retest reliabilty, construct validity (cross-informant agreement) and external validity, where we calculated the correlation of the number of differenet traumatic events with psychopathological symptoms.Methods: 280 child-parent pairs (children aged 11.3 ± 2.2 years) from various Slovenian primary schools participated in the study. They were divided into two groups: 180 healthy primary school students and 100 children with Type 1 Diabetes (our study was a part of a larger study The Influence of Psychobiological Adversity on Children and Adolescents with Type 1 Diabetes Study). Two versions of the LITE questionnaire were used. Children completed the child report (LITE-S) and parents the parent report (LITE-P) version. After 4 weeks, 117 children, and 114 parents filled out the LITEs again. External validity was assessed using the Youth Self Report and Child Behaviour Checklist syndrome-oriented scales.Results: Retest reliability for individual scales was r = 0.469–0.639 (ρ = 0.443–0.636; p < 0.001), but higher for individual items (κ = 0.263–0.821; p < 0.001). Correlations between reports from parents and children were r = 0.313–0.345 (ρ = 0.317–0.348; p < 0.001). The number of different events experienced by children correlated significantly with the measured depressive—anxiety, and posttraumatic stress disorder symptoms.Conclusions: Based on our results, the LITE-S and LITE-P “All events” scale have acceptable psychometric properties for use in research and in clinical practise screening. We recommend looking at single items, taking into consideration the responses from both the child and the parent for more precise information. To improve the precision of the psychodiagnostic capacity of the questionnaire, further research on various populations should be performed.

Background/Objective: Type 1 diabetes (T1D) is among the most common chronic diseases in children/adolescents, and the incidence continues to rise worldwide. Different environmental factors have been evaluated in the etiology. In the present study, we investigated the role of attachment examining whether insecure attachment to carers or carers' own attachment insecurity was related to a higher risk of T1D in children.Methods: We included 101 children with T1D (mean age 11.8 years), 106 healthy controls (11.6 years), and one of their carers. We assessed children's attachment using the Child Attachment Interview and carers' attachment using the Relationship Structures Questionnaire. We constructed binary multinomial logistic regression models using attachment to mothers, carers' attachment representations, and stressful life-events as T1D predictors.Results: Higher carer attachment anxiety was associated with the child's T1D diagnosis (p < 0.05; R2 = 0.0613) while security of attachment to mothers showed no significant association. When mothers' education was included in the model, both attachment anxiety in higher educated mothers and stressful life events showed a significant association with the child's T1D (p < 0.001; R2 = 0.293).Conclusions: Our findings suggest that higher attachment-related anxiety in carers with high education and stressful life events are associated with T1D in children.

For the purpose of the morphometric characterization and differentiation of local sheep breeds that belong to the group of breeds called Pramenka or Zackel, two Slovenian (Bela Krajina and Istrian Pramenka) and four Montenegrin sheep breeds (Bardoka, Sjenička, Pivska Pramenka, and Zeta Žuja) were studied. The variation of morphometric measures and nine morphometric indices were analysed. Principal component analysis (PCA) was applied in order to provide an easier description of body size and shape. Regarding body size, the Sjenička breed was one of the largest breeds (body weight 76.4 kg, wither height 72.7 cm, chest circumference 100.3 cm), while Zeta Žuja had the smallest body size (37.1 kg, 64.8, and 81.9 cm). On the other hand, Slovenian Istrian Pramenka had the largest body length, chest depth, chest width, and rump width among all included breeds (79.4, 33.6, 22.7, and 21.2 cm). Bela Krajina, Istrian Pramenka, and the Sjenička breed, according to the index of body frame (IBF) value (107–114), have a rectangular body frame, Bardoka and Pivska Pramenka have a square body frame (99.3–100), and Zeta Žuja has a short body frame (91.8). The PCA of all morphometric parameters extracted three components accounting for 96.6 % of the cumulative variance. An unweighted pair–group method with arithmetic mean (UPGMA) cluster analysis by Euclidian distance shows diversity among the studied breeds, through it grouped Pivska Pramenka with Sjenička and Istrian with Bela Krajina Pramenka in two clusters, while Bardoka and Zeta Žuja were clustered separately.

Abstract. Mycobacteria belonging to Mycobacterium (M.) avium complex (MAC) and M. abscessus complex (MABSC) are the most frequent causes of mycobacteriosis in the world. In the last few years MAC and MABSC taxonomy was rapidly changing due to new molecular methods conveying the possibility to differentiate between species. New techniques are able to identify M. chimaera that was previously recognized as M. intracellulare and also differentiate subspecies of MABSC. Due to their natural habitat, non-tuberculous mycobacteria (NTM) are constantly exposed to various concentrations of antimicrobial drugs and other chemicals and consequently they had developed different mechanisms of resistance. Macrolides and aminoglycosides are frequently used drugs to treat MAC and MABSC infections. The aim of our nation-wide survey was to obtain information about MABSC subspecies prevalence in Slovenia and to assess the percentage of misidentifications of M. chimaera isolates as M. intracellulare in the past. Moreover, the purpose of our study was to reveal, which of the two species M. intracellulare or M. chimaera is clinically more relevant in Slovenia. Further, the aim of the study was to detect mutations in erm(41), rrl and rrs genes, which are known to convey macrolide resistance (erm(41) and rrl) and aminoglycoside resistance (rrs). One hundred and thirty-two Slovenian mycobacterial isolates obtained from the National Mycobacterial Collection that belong to MAC and MABSC were analysed. GenoType NTM-DR was used to differentiate M. intracellulare from M. chimaera and subspecies of MABSC. Our results showed that 48% of previously identified M. intracellulare isolates were actually M. chimaera isolates and that M. abscessus subsp. abscessus was the most frequent subspecies of MABSC. Most of the MABSC isolates carried the inducible macrolide resistance genes (erm(41) and rrl), however none of the isolates of MAC and MABSC had mutations in rrs genes for aminoglycoside resistance.

Here we present the first welfare assessment of commercial layers conducted in Slovenia. Hens were assessed in four systems at the beginning of the laying period at 22 to 24 weeks and at 50 to 55 weeks of age. These systems were an enriched battery cage system, an aviary, and a litter system with or without outdoor access. Clinical inspections of flocks were performed, and animal-based welfare indicators were scored (e.g., keel bone damage, feather condition, foot pad lesions, beak deformities, and comb and skin wounds). Hens’ fear level was scored using the novel object test and avoidance distance test. Among resource-based measures, selected micro-climate parameters were measured. The results showed no obvious clinical signs related to infectious diseases and suggest that the selected climate conditions were satisfying in all systems. Among animal-based welfare indicators, keel bone damage was shown to be the most serious problem connected with hens’ age and housing systems (p < 0.05). Enriched cages and aviary system were associated with significantly more keel deformities compared to the litter systems (p < 0.05). In addition, the least prevalence of foot pad dermatitis together with better feather condition was observed in the litter systems. In the family-owned aviary facility, hens were found to be the most motivated to approach a novel object or a human, and as such were recognized as the least fearful birds, with better human–animal interaction compared to other intensive housing systems. Key words: laying hens; welfare; health; housing system OCENA DOBROBITI V INTENZIVNIH REJAH KOKOŠI NESNIC V SLOVENIJI Povzetek: Opravili smo prvo celovito oceno dobrobiti kokoši nesnic v Sloveniji. V raziskavo smo vključili nesnice iz štirih različnih sistemov reje in raven dobrobiti ocenili v dveh starostnih obdobjih; na začetku nesnosti, v starosti od 22 do 24 tednov in pri 50 do 55 tednih. Nesnice so bile rejene v obogatenih kletkah, v voljerah, v talni reji brez možnosti izpusta in v talni reji z možnostjo izpusta. Ob vsakem ocenjevanju smo jate klinično pregledali in s pregledom posameznih živali ocenili specifične indikatorje dobrega počutja (poškodbe prsnice, operjenost, poškodbe podplatnih blazinic, deformacije kljuna in poškodbe grebena ter kože). Plašnost kot indikator socialnega obnašanja smo ocenili s testom novega predmeta in s testom odmika od človeka. Spremljali smo tudi mikro-klimatske pogoje reje. Ves čas spremljanja nismo ugotovili vidnih kliničnih znakov kužnih obolenj. Rezultati meritev mikro-klimatskih parametrov nakazujejo, da so bili pogoji v rejah dobri. Poškodba prsnice se je izmed specifičnih kazalnikov izkazala za najresnejši problem, na katerega vplivata tako starost kot sistem reje (p < 0,05). Poškodbe prsnice so bile značilno bolj izražene pri kokoših iz obogatenih kletk in voljer (p < 0,05) v primerjavi z nesnicami iz talnih rej. Kokoši iz talnih sistemov so bile tudi boljše operjene in so imele nižjo prevalenco poškodb na podplatnih blazinicah. Nesnice, ki so bile rejene v voljerah na družinski kmetiji, so kazale največ zanimanja za nove predmete in človeka. Te kokoši so bile ocenjene kot najmanj plašne in so izražale boljšo interakcijo človek–žival kot kokoši iz drugih primerjanih sistemov. Ključne besede: kokoši nesnice; dobrobit; zdravje; sistem reje

ABSTRACT A selection of commercially available antibodies, targeted against markers employed in studies of mammary gland biology, was tested to determine their reactivity in goat mammary tissue and the derived tissue cultures. Expression of the markers smooth muscle actin ( SMA ), selected keratins ( KRT ) 5, 14, 18 , and 19 , CD24 molecule ( CD24 ), epithelial cell adhesion molecule ( EPCAM ), mucin 1 ( MUC1 ), integrin subunit alpha 6 ( ITGA6; CD49F ), integrin subunit beta 1 ( ITGB1; CD29 ), cyclin dependent kinase inhibitor 1A ( CDKN1A; p21 ), membrane metalloendopeptidase ( MME; CD10 ), progesterone receptor ( PGR ), estrogen receptor 1 ( ESR1 ), and vimentin ( VIM ) was first assessed on mRNA level, using reverse transcription PCR (RT-PCR). The reactivity of the antibodies in the tissue sections and the derived tissue cultures was determined using immunofluorescence. The result of this study is a list of commercially available antibodies, raised mostly against human antigens, which also recognize orthologous goat antigens and are useful for characterization of different mammary cell types. Additionally, primers that are functional in detecting expression of mammary lineage markers in goat mammary mRNA isolates were validated. The suggested antibodies, PCR primers, and the described methods are of practical value for researchers interested in characterization and isolation of cell types comprising mammary tissue of goats and probably other ruminants.

Extracellular vesicles with their molecular cargo can modulate target cell response and may affect the pathogenesis of diseases. The extracellular vesicles containing micro-RNAs (miRNAs), which are often studied as disease biomarkers, but rarely as mediators of the disease development. The role of extracellular vesicles derived miRNAs in type 1 diabetes is currently not well established. We observed a fraction of blood plasma extracellular vesicles positive for membrane proteins potentially associated with insulin-producing beta-cells and identified differentially expressed extracellular vesicles derived miRNAs in individuals with type 1 diabetes. These differentially expressed extracellular vesicles derived human miRNAs in participants with type 1 diabetes and participants with Langerhans islets beta-cells destruction showed the ability to activate TLR7/8 signaling cascade and increase activation as well as cytotoxicity of the effector blood immune cells with cytokine and chemokine release. Our results illustrate extracellular vesicles derived human miRNAs as modulators of the immune system in type 1 diabetes autoimmunity, providing potentially new insight into the pathogenesis of the disease, and novel molecular targets for intervention and type 1 diabetes prevention.

Only very recently, has it been proposed that the hitherto existing Mycobacterium kansasii subtypes (I–VI) should be elevated, each, to a species rank. Consequently, the former M. kansasii subtypes have been denominated as Mycobacterium kansasii (former type I), Mycobacterium persicum (II), Mycobacterium pseudokansasii (III), Mycobacterium innocens (V), and Mycobacterium attenuatum (VI). The present work extends the recently published findings by using a three-pronged computational strategy, based on the alignment fraction-average nucleotide identity, genome-to-genome distance, and core-genome phylogeny, yet essentially independent and much larger sample, and thus delivers a more refined and complete picture of the M. kansasii complex. Furthermore, five canonical taxonomic markers were used, i.e., 16S rRNA, hsp65, rpoB, and tuf genes, as well as the 16S-23S rRNA intergenic spacer region (ITS). The three major methods produced highly concordant results, corroborating the view that each M. kansasii subtype does represent a distinct species. This work not only consolidates the position of five of the currently erected species, but also provides a description of the sixth one, i.e., Mycobacterium ostraviense sp. nov. to replace the former subtype IV. By showing a close genetic relatedness, a monophyletic origin, and overlapping phenotypes, our findings support the recognition of the M. kansasii complex (MKC), accommodating all M. kansasii-derived species and Mycobacterium gastri. None of the most commonly used taxonomic markers was shown to accurately distinguish all the MKC species. Likewise, no species-specific phenotypic characteristics were found allowing for species differentiation within the complex, except the non-photochromogenicity of M. gastri. To distinguish, most reliably, between the MKC species, and between M. kansasii and M. persicum in particular, whole-genome-based approaches should be applied. In the absence of clear differences in the distribution of the virulence-associated region of difference 1 genes among the M. kansasii-derived species, the pathogenic potential of each of these species can only be speculatively assessed based on their prevalence among the clinically relevant population. Large-scale molecular epidemiological studies are needed to provide a better understanding of the clinical significance and pathobiology of the MKC species. The results of the in vitro drug susceptibility profiling emphasize the priority of rifampicin administration in the treatment of MKC-induced infections, while undermining the use of ethambutol, due to a high resistance to this drug.

MicroRNAs (miRNA) are a class of non-coding RNAs important in posttranscriptional regulation of target genes. Regulation requires complementarity between the target mRNA and the miRNA region responsible for their recognition and binding, also called the seed region. Previous studies have proven that expression profiles and genetic variations of miRNA genes (miR-SNP; SNP – single nucleotide polymorphism) and their target sites (miR-TS-SNPs) have an impact on phenotypic variation and disease susceptibility in human, animal models, and livestock. MicroRNA-associated polymorphisms therefore represent biomarker potential for phenotypic traits in livestock. Effects of miRNA gene polymorphisms on phenotypic traits have been studied in several animal species but much less in cattle. The aim of the present study was therefore to analyze the genetic variability in the bta-mir-2313 gene and test associations with growth and carcass traits of the Slovenian Simmental cattle breed. Additionally, validated and predicted genomic information related to the miRNA gene bta-mir-2313 has been obtained and presented as an atlas of miRNA regulatory elements. Sanger sequencing has been used for biomarker development and genotyping of 145 animals of Slovenian dual-purpose Simmental cattle. Out of nine known polymorphisms located within pre-miRNA regions, one mature miRNA seed SNP was polymorphic in the Slovenian Simmental cattle breed. An additional three polymorphisms were identified within the flanking pri-miRNA regions. There was no significant effect of polymorphisms on 18 tested fattening and carcass traits; however, validated polymorphisms could now be tested in association with other traits in other cattle populations. The microRNA gene bta-mir-2313 warrants further genetic and functional analyses since it overlaps with a large number of quantitative trait loci (QTL), has over 3100 predicted targets and highly polymorphic mature seed regions, and is located within protein-coding gene GRAMD1B, previously associated with production traits in cattle. Mature miRNA seed SNPs present important genomic loci for functional studies because they could affect the gain/loss of downstream targets and should be systematically studied in cattle.

Mitochondrial transcription factor A plays, due to its role in replication and transcription of the mitochondrial DNA, an important role in mitochondrial biogenesis and regulation of its activity. In different mammalian species, mitochondrial transcription factor A is present in two different splicing forms. The aim of the study was to verify the presence of both splicing forms and to estimate their ratio in different tissues. The expression levels of both splicing forms in two skeletal muscles (m. semispinalis capitis and m. semimembranosus), heart muscle, brain, spleen, and liver in commercial hybrids and in the autochthonous Krsko polje pig breed are reported. In all analyzed tissues the expression of the long form of the mitochondrial transcription factor A was about one order of magnitude higher compared to the short form, lacking almost the entire exon 4 region. Taking into account the expression ratio of both splicing forms and biological function of the mitochondrial transcription factor A, the possible explanation for the persistence of both forms in different mammalian species is provided.

Istrian goat is an autochthonous Croatian breed which inhabited Istrian peninsula and was important in milk production and human nutrition, especially for poor people. For centuries Istrian goat was a recognizable heraldic symbol of Istria, but in her real form almost disappeared from the breeding area. The revitalization and reaffirmation of Istrian goat began with several dozen remaining breeding animals, after a decade-long breeding ban. Genetic characterization of the Istrian goat population is necessary for providing insight into the state of genetically preserved structure within population as well as positioning Istrian goats within phylogenetically related breeds. Microsatellite and mtDNA analysis of reproductive individuals of Istrian goat and related breeds, Croatian White goat and Saanen goat was carried out. In the population of Istrian goat, higher allelic variability (nA = 9.7; AR = 7.4) were found as well as significant genetic distance (FST = 0.068 - 0.086) in relation to other two breeds. Such results indicated that Istrian goat constitutes a separate genetic identity. The observed ten haplotype sequences of the D-loop mtDNA also confirm the significant genetic richness of the maternal hereditary component. The observed haplotypes in the population of Istrian goat belong to lineage A. A smaller number of haplotypes shows similarity to the group of “white” goats, indicating traces of earlier limited but targeted crossing of Istrian goats. The genetic profile analysis of Istrian goats indicates a high level of genetic variability and provides guidelines for a long-term conservation program. The preserved genetic and promising potential of milk production of Istrian goat makes a significant basis for her economic reaffirmation. Orientation of the breed towards milk production could be an efficient strategy for its effective preservation.

The study was conducted between March and September 2019 in six meat-type turkey flocks with similar management standard procedures using the transect walk method. The concept of the method is based on visual observation of the birds while slowly walking across the entire farm in predetermined transects. Each flock was evaluated at three different times during the fattening cycle: at 3 to 4, 12 to 13, and 19 to 20 weeks of age, and total number of males and females that were immobile or lame, had visible head, vent, or back wounds, were small, featherless, dirty, or sick, had pendulous crop, or showed aggression toward birds or humans were recorded. At each visit, NH3 and CO2 were measured within the facilities. In the first assessment, the most frequently observed welfare indicators were small size (0.87%) and immobility (0.08%). Males showed a significantly higher prevalence of small size (p < 0.01), sickness (p < 0.05), and dirtiness (p < 0.1) compared to females. In the second assessment, the most common findings in both sexes were dirtiness (1.65%) and poor feather condition (1.06%), followed by immobility (0.28%). Males were significantly dirtier (p < 0.001), had more immobile birds (p < 0.01) and birds with vent wounds (p < 0.1), but had fewer sick birds (p < 0.05). In the last assessment, an increase in immobile, lame, sick, and dead birds was recorded, indicating an increase in health problems. Higher CO2 (3000 and 4433 ppm) and NH3 (40 and 27.6 ppm) values were noted only at the first assessment in two facilities. Further analyses showed that slightly elevated NH3 and CO2 levels did not influence the occurrence of welfare indicators. This study is the first description of the welfare of commercial turkey flocks in Slovenia.