Mastitis is one of the most frequently encountered diseases in dairy cattle, negatively affecting animal welfare and milk production. For this reason, contributions to understanding its genomic architecture are of great interest. Genome-wide association studies (GWAS) have identified multiple loci associated with somatic cell score (SCS) and mastitis in cattle. However, most of the studies have been conducted in different parts of the world on various breeds, and none of the investigations have studied the genetic architecture of mastitis in Romanian dairy cattle breeds up to this point in time. In this study, we report the first GWAS for SCS in dairy cattle breeds from Romania. For GWAS, we used an Axiom Bovine v3 SNP-chip (>63,000 Single Nucleotide Polymorphism -SNPs) and 33,330 records from 690 cows belonging to Romanian Spotted (RS) and Romanian Brown (RB) cattle. The results found one SNP significantly associated with SCS in the RS breed and 40 suggestive SNPs with −log₁₀ (p) from 4 to 4.9 for RS and from 4 to 5.4 in RB. From these, 14 markers were located near 12 known genes (AKAP8, CLHC1, MEGF10, SATB2, GATA6, SPATA6, COL12A1, EPS8, LUZP2, RAMAC, IL12A and ANKRD55) in RB cattle, 3 markers were close to ZDHHC19, DAPK1 and MMP7 genes, while one SNP overlapped the HERC3 gene in RS cattle. Four genes (HERC3, LUZP2, AKAP8 and MEGF10) associated with SCS in this study were previously reported in different studies. The most significant SNP (rs110749552) associated with SCS was located within the HERC3 gene. In both breeds, the SNPs and position of association signals were distinct among the three parities, denoting that mastitis is controlled by different genes that are dependent according to parity. The current results contribute to an expansion in the body of knowledge regarding the proportion of genetic variability explained by SNPs for SCS in dairy cattle.

Considering the major limitations of the latest studies conducted in Romania on the knowledge, attitudes and practices (KAPs) of antibiotic use and antibiotic resistance, we conducted this study to assess this major public health threat. A cross-sectional survey based on a validated questionnaire was conducted among the general population of Romania for a period of 5 months, i.e., September 2021–January 2022. The questionnaire was distributed using Google Form and it covered demographic characteristics and KAP assessments consisting of 12 items on knowledge, 10 items on attitudes and 3 items on practices. Latent class analyses (LCAs) were conducted to group respondents based on their responses. The response rate was 77%, of which females responded in a greater number (n = 1251) compared to males (n = 674). For most of the respondents (67.32%, n = 1296), the education level was high school, while 23.58% (n = 454) of respondents were college graduates. One in three Romanians (33.3%) know the WHO predictions related to this topic. Overall, the Romanian population is less disciplined when it comes to completing antibiotic treatments, as 29.19% of the respondents stop the course of antibiotic administration if their symptoms improve. The key findings from the present study may help policy makers in designing targeted interventions to decrease confusion, ambiguity or misconceptions about antibiotic use.

Yeast infections are often connected with formation of biofilms that are extremely difficult to eradicate. An excellent model system for deciphering multifactorial determinants of yeast biofilm development is the colony biofilm, composed of surface (“aerial”) and invasive (“root”) cells. While surface cells have been partially analyzed before, we know little about invasive root cells. In particular, information on the metabolic, chemical and morphogenetic properties of invasive versus surface cells is lacking. In this study, we used a new strategy to isolate invasive cells from agar and extracellular matrix, and employed it to perform genome wide expression profiling and biochemical analyses of surface and invasive cells. RNA sequencing revealed expression differences in 1245 genes with high statistical significance, indicating large genetically regulated metabolic differences between surface and invasive cells. Functional annotation analyses implicated genes involved in stress defense, peroxisomal fatty acid β-oxidation, autophagy, protein degradation, storage compound metabolism and meiosis as being important in surface cells. In contrast, numerous genes with functions in nutrient transport and diverse synthetic metabolic reactions, including genes involved in ribosome biogenesis, biosynthesis and translation, were found to be important in invasive cells. Variation in gene expression correlated significantly with cell-type specific processes such as autophagy and storage compound accumulation as identified by microscopic and biochemical analyses. Expression profiling also provided indications of cell-specific regulations. Subsequent knockout strain analyses identified Gip2p, a regulatory subunit of type 1 protein phosphatase Glc7p, to be essential for glycogen accumulation in surface cells. This is the first study reporting genome wide differences between surface and invasive cells of yeast colony biofilms. New findings show that surface and invasive cells display very different physiology, adapting to different conditions in different colony areas and contributing to development and survival of the colony biofilm as a whole. Notably, surface and invasive cells of colony biofilms differ significantly from upper and lower cells of smooth colonies adapted to plentiful laboratory conditions.

In the present study, we investigated one polymorphism of the PRL gene (rs211032652 SNP) and assessed its influence on milk production and chemical composition in two Romanian cattle breeds. A total of 119 cattle from two breeds reared in Western Romania (64 Romanian Spotted and 55 Romanian Brown) were included in the research herd. A PCR-RFLP genotyping assay was used for the identification of the rs211032652 SNP variants. Shapiro’s test and Levene’s test were used to verify ANOVA assumptions and ANOVA and Tukey’s test were employed to test the associations between PRL genotypes and five milk traits. Among the studied breeds, our results showed that PRL genotypes were significantly associated (p < 0.05) with fat and protein percentage in the milk of Romanian Brown cattle. The AA genotype was associated with a higher fat percentage in milk (4.76 ± 0.28) compared to the GG genotype (4.04 ± 0.22, p = 0.048), as well as a higher protein percentage (3.96 ± 0.32% vs. 3.43 ± 0.15%, p = 0.027) in Romanian Brown cattle. Moreover, the PRL locus favored a significantly higher fat (p = 0.021) and protein (p = 0.028) percentage in the milk of Romanian Brown cattle compared to the Romanian Spotted breed, with a difference of 0.263% and 0.170%, respectively.

Mastitis is the most common production disease in the dairy sector worldwide, its incidence being associated with both cows’ exposure to bacteria and the cows’ genetic make-up for resistance to pathogens. The objective of our study was to analyse 89 missense SNPs belonging to six genes (CXCR2, CXCL8, TLR4, BRCA1, LTF, BOLA-DRB3), which were found to be associated with genetic resistance or susceptibility to mastitis. A total of 298 cattle (250 Romanian Spotted and 48 Romanian Brown) were genotyped by Kompetitive Allele Specific PCR (KASP) and a chi-squared test was used for genetic association studies with clinical mastitis. A total of 35 SNPs (39.3%) among the selected 89 SNPs were successfully genotyped, of which 31 markers were monomorphic. The polymorphic markers were found in two genes: TLR4 (rs460053411) and BOLA-DRB3 (rs42309897, rs208816121, rs110124025). The polymorphic SNPs with MAF > 5% and call rates > 95% were used for the association study. The results showed that rs110124025 in the BOLA-DRB3 gene was significantly associated with mastitis prevalence (p ≤ 0.05) in both investigated breeds. Current results show that the SNP rs110124025 in the BOLA-DRB3 gene can be used as a candidate genetic marker in selection for mastitis resistance in Romanian dairy cattle.

Milk protein content represents a key economic trait in dairy production, yet the genetic architecture underlying this trait remains unexplored in Romanian dual-purpose cattle breeds. This study conducted a genome-wide association analysis for milk protein content in 313 Romanian Simmental (n = 271) and Romanian Brown (n = 42) cows belonging to the Research and Development Station for Bovine Arad, Romania. Following quality control, 33,531 SNPs were tested for association with protein percentage adjusted for other effects (breed, days in milk, season, year, parity) using linear regression with the first five principal components as covariates to control population stratification. Although no SNP reached genome-wide significance (p < 5 × 10⁻⁸), one SNP achieved significance (p < 2.98 × 10⁻⁵) and seven additional SNPs exceeded the nominal threshold (p < 1 × 10⁻⁴) across six chromosomes. The strongest association (p = 9.56 × 10⁻⁶) mapped to chromosome 25 near C7orf61. Biologically relevant candidate genes included KLF6 on chromosome 13, previously associated with milk traits in Chinese Holstein, and AHCYL1 on chromosome 3, involved in calcium homeostasis. These findings provide initial insights into genomic regions influencing milk protein content in Romanian dual-purpose cattle, though validation in larger cohorts needs to be carried out.