Corpus luteum (CL) is an endocrine structure that undergoes substantial changes over short intervals during the bovine reproductive (estrous) cycle. These changes are regulated by a vast array of signaling molecules, mainly proteins, that govern reproduction. We collected a comprehensive set of CLs from non-pregnant (days 1-2, 3-4, 5-7, 8-12, 13-17, and >18) and pregnant (months 1-2, 3-4, 5-6, and >7) cows, enabling a detailed characterization of temporal changes in CL biology. While previous studies have focused on a limited number of proteins, we employed a broader approach to exploring additional proteins potentially involved in CL regulation across different stages. Using high-resolution mass spectrometry (HR-LC-MS/MS), we identified approximately 3,783 individual proteins. These proteins may be useful for research in species where access to well-characterized CL tissue is limited. Our findings indicate diverse changes in protein expression across groups, highlighting proteomic patterns that may reflect key processes of bovine reproduction, particularly during the transition toward regression, a critical period that influences whether the CL is maintained to support pregnancy or undergoes regression.

The purpose of the study was to characterize the expression patterns of apelin and its receptor APJ (apelin/APJ) in superovulation induced follicles and corpus luteum (CL) tissue in the cow. Ovaries containing different classes of preovulatory follicles and early CL were timely defined during GnRH (gonadotropin hormone-releasing hormone) induced superovulation as follows: Before GnRH application, follicles group I: 0 h (control group) and after GnRH; group II: 4 h (corresponds to LH peak); group III:10 h; group IV: 20 h; group V: 25 h (corresponds close to ovulation) and group VI: 60 h (after ovulation - early CL). This experiment has made it possible for the direct comparison of the apelin/APJ system members in follicle tissue (theca interna plus granulosa cells) of periovulatory follicle groups with the subsequent CL tissue after ovulation (early CL, day 2-3). Relative gene expression levels were measured by RT-qPCR. The low level of apelin mRNA in the follicle group before the GnRH application (Group I), remained unchanged even in the follicle group II and III (4 h and 10 h after GnRH respectively). Apelin expression levels increased significantly by only 20 h after GnRH (group IV follicles: just before ovulation), remaining high during follicle ovulation (Group V: 20 h) and after ovulation (Group VI: early CL). In contrast, the low APJ mRNA level increased significantly in follicle group II (during LH peak) remaining high in all follicle groups before ovulation, as well as after ovulation (early CL). Our results indicate the possible involvement of apelin system (apelin/APJ) as a possible key regulator in the local mechanisms supporting final follicle maturation and ovulation, as well as the follicular-luteal transition and CL formation in cows.

The objective of the study was to characterize the mRNA expression patterns of specific steroid hormone receptors namely, estrogen receptors (ESRRA-estrogen related receptor alpha and ESRRB-estrogen related receptor beta) and progesterone receptors (PGR) in superovulation-induced bovine follicles during the periovulation and subsequent corpus luteum (CL) formation. The bovine ovaries (n = 5 cow / group), containing preovulatory follicles or early CL, were collected relative to injection of the gonadotropin-releasing hormone (GnRH) at (I) 0 h, (II) 4 h, (III) 10 h, (IV) 20 h, (V) 25 h (preovulatory follicles) and (VI) 60 h (CL, 2-3 days after induced ovulation). In this experiment, we analyzed the steroid receptor mRNA expression and their localization in the follicle and CL tissue. The high mRNA expression of ESRRA, ESRRB, and PGR analyzed in the follicles before ovulation is significantly reduced in the group of follicles during ovulation (25 h after GnRH), rising again significantly after ovulation in newly formed CL, only for ESRRA and PGR (P < 0.05). Immunohistochemically, the nuclei of antral follicles' granulosa cells showed a positive staining for ESRRA, followed by higher activity in the large luteal cells just after ovulation (early CL). In contrast, the lower PGR immunopresence in preovulatory follicles increased in both small and large luteal cell nuclei after follicle ovulation. Our results of steroid receptor mRNA expression in this experimentally induced gonadotropin surge provide insight into the molecular mechanisms of the effects of steroid hormones on follicular-luteal tissue in the period close to the ovulation and subsequent CL formation in the cow.

The ubiquitous presence and potent carcinogenic effects of aflatoxins, a group of mycotoxins, pose a significant threat to global food safety, especially within the dairy sector. This research aimed to evaluate the prevalence and distribution of Aflatoxin M1 in diverse cheese varieties within Kosovo’s markets in 2022 and discern potential correlations with cheese types, processing conditions, and nutritional parameters to enhance understanding and ensure the safety of dairy products. In the latter half of 2022, a total of 93 different types of manufactured cheese samples from 15 different countries were analysed for Aflatoxin M1 toxin content using the enzyme-linked immunosorbent assay technique. The results of the research indicate that several countries have exceeded the concentration of 0.05 ug/kg for Aflatoxin M1, set as the maximum tolerable limit for milk and milk products in many countries. Notable, cheese samples from Albania exhibited substantially (p=0.000) higher levels of Aflatoxin M1 compared to other nations. The results showed that the Aflatoxin M1 concentration did not exhibit statistically significant variations based on the regulated total fat content (p=0.902), carbohydrate content (p=0.741), protein concentration (p=0.298), or salt content (p=0.122) within the cheese. The results demonstrated that Aflatoxin M1 levels were not significantly (p=0.926) affected by cheese consistency. While different starter cultures were associated with varying Aflatoxin M1 concentrations, these differences were not statistically significant (p=0.472). Similarly, ageing time did not exert a significant (p=0.691) impact on AFM1 levels, as concentrations remained relatively consistent across different ageing periods. These findings provide valuable insights into the variations in Aflatoxin M1 levels in cheese samples among different countries. They highlight the importance of regulatory measures and continuous monitoring to ensure food safety and adherence to maximum allowable limits

The objective of the study was to characterise the expression patterns of the two key components of cortisol action namely HSD11B1 (11-beta-hydroxysteroid dehydrogenase type 1) and NR3C1 (nuclear receptor subfamily 3, group C, member 1, also known as the glucocorticoid receptor) in superovulation induced bovine follicles during the periovulation and subsequent corpus luteum (CL) formation. Bovine ovaries containing preovulatory follicles or CL were timely defined during induced ovulation as follows: 0 h before GnRH (Gonadotropin-releasing hormone) application, and 4, 10, 20, 25 (follicles) and 60 h (early CL) after GnRH. The low mRNA expression of HSD11B1 and NR3C1 in the follicle group before the GnRH application increased significantly in the follicle group 20 h after GnRH and remained high afterward also in the early CL group. In contrast, the high NR3C1 mRNA decreased in follicles 25 h after GnRH (close to ovulation) and significantly increased again after ovulation (early CL). Our results indicated the involvement of HSD11B1 and NR3C1 as the two key components of cortisol action in the local mechanisms coordinating final follicle maturation, ovulation, follicular-luteal transition and CL development in the cow.

The study aimed to assess the local gene expression of adipokine members, namely vaspin, adiponectin, visfatin, resistin and their associated receptors - heat shock 70 protein 5 (HSPA5), adiponectin receptor 1 (AdipoR1) and adiponectin receptor 2 (AdipoR2) - in bovine follicles during the preovulatory period and early corpus luteum development. Follicles were collected before gonadotropin-releasing hormone (GnRH) treatment (0 h) and at 4, 10, 20, 25 and 60 h after GnRH application through transvaginal ovariectomy (n = 5 samples/group). Relative mRNA expression levels were quantified using real-time reverse transcription polymerase chain reaction (RT-qPCR). Vaspin exhibited high mRNA levels immediately 4 h after GnRH application, followed by a significant decrease. Adiponectin mRNA levels were elevated at 25 h after GnRH treatment. AdipoR2 exhibited late-stage upregulation, displaying increased expression at 20, 25 and 60 h following GnRH application. Visfatin showed upregulation at 20 h post-GnRH application. In conclusion, the observed changes in adipokine family members within preovulatory follicles, following experimentally induced ovulation, may constitute crucial components of the local mechanisms regulating final follicle growth and development.

The study aimed to evaluate the mRNA expression levels of various local novel adipokines, including vaspin, adiponectin, visfatin, and resistin, along with their associated receptors, heat shock 70 protein 5, adiponectin receptor 1, and adiponectin receptor 2, in the corpus luteum (CL) during luteal regression, also known as luteolysis, in dairy cows. We selected Fleckvieh cows in the mid-luteal phase (days 8-12, control group) and administered cloprostenol (PGF analog) to experimentally induce luteolysis. We collected CL samples at different time points following PGF application: before treatment (days 8-12, control group) and at 0.5, 2, 4, 12, 24, 48, and 64 h post-treatment (n = 5) per group. The mRNA expression was measured via real-time reverse transcription polymerase chain reaction (RT-qPCR). Vaspin was characterized by high mRNA levels at the beginning of the regression stage, followed by a significant decrease 48 h and 64 h after PGF treatment. Adiponectin mRNA levels were elevated 48 h after PGF. Resistin showed upregulation 4 h post PGF application. In summary, the alterations observed in the adipokine family within experimentally induced regressing CL tissue potentially play an integral role in the local regulatory processes governing the sequence of events culminating in functional luteolysis and subsequent structural changes in the bovine ovary.

Aflatoxin M1 (AFM1), a toxic byproduct of aflatoxin B1 (AFB1) produced by Aspergillus fungi, is a carcinogenic mycotoxin that can contaminate various agricultural commodities. It can be transferred from AFB1-contaminated feed to milk and dairy products, including yogurt, posing a potential health risk to consumers. In spring 2023, a total of 74 yogurt samples were collected from the largest food suppliers in Kosovo for analysis, including samples produced in Kosovo and seven other countries: Albania, North Macedonia, Bosnia and Herzegovina, Slovenia, Greece, Italy, and Germany. A rapid and sensitive analytical method, Enzyme-linked immunosorbent assay (ELISA), was used for the analysis. The results of the study highlight discernible differences in the maximum tolerable levels of AFM1 between the countries. More specifically, yogurt samples from Slovenia and Germany had lower levels than those from other countries. Additionally, the median levels of AFM1 in samples from Slovenia and Germany were significantly lower. The mean concentrations of AFM1 in yogurt samples from Kosovo and other countries were 0.071 μg/kg and 0.080 μg/kg, respectively. Out of all samples, 66 (89%) exceeded the maximum tolerable limit of 0.05 μg/kg. Among the exporting countries, Albania had the highest median AFM1 level of 0.085 μg/kg and the highest maximum level of 0.195 μg/kg. Slovenia had the lowest median AFM1 level, while Germany had the lowest maximum AFM1 level. All samples from Albania, Greece, and Bosnia and Herzegovina exceeded the maximum tolerable limit. High prevalence was also observed in samples from Kosovo, North Macedonia, and Slovenia. Considering the average daily consumption of about 250 grams of yogurt, and the total median value of Aflatoxin M1 concentration (0.071 μg/kg), the estimated daily intake was calculated to be 0.017 μg. These findings highlight the importance of monitoring and enforcing regulatory limits to ensure yogurt safety and to protect public health. Efforts should be focused on mitigating AFM1 contamination and implementing measures to minimise its presence in dairy products, especially in regions where levels exceed the established limits.

Prostaglandins are synthesized from arachidonic acid through the catalytic activities of cyclooxygenase, while the production of different prostaglandin types, prostaglandin F2 alpha (PGF) and prostaglandin E2 (PGE), are regulated by specific prostaglandin synthases (PGFS and PGES). Prostaglandin ligands (PGF and PGE) bind to specific high-affinity receptors and initiate biologically distinct signalling pathways. In the ovaries, prostaglandins are known to be important endocrine regulators of female reproduction, in addition to maintaining local function through autocrine and/or paracrine effect. Many research groups in different animal species have already identified a variety of factors and molecular mechanisms that are responsible for the regulation of prostaglandin functions. In addition, prostaglandins stimulate their intrafollicular and intraluteal production via the pathway of prostaglandin self-regulation in the ovary. Therefore, the objective of the review article is to discuss recent findings about local regulation patterns of prostaglandin ligands PGF and PGE during different physiological stages of ovarian function in domestic ruminants, especially in bovine. In conclusion, the discussed local regulation mechanisms of prostaglandins in the ovary may stimulate further research activities in different methodological approaches, especially during final follicle maturation and ovulation, as well as corpus luteum formation and function.

This study aimed to determine the gene expression of different local novel adipokines, such as vaspin, adiponectin, visfatin, and resistin, and their known receptors, namely, heat shock 70 protein 5, adiponectin receptor 1, and adiponectin receptor 2, in the bovine corpus luteum (CL) during different phases of the estrous cycle (on days 1–2, 3–4, 5–7, 8–12, 13–18, >18) and pregnancy (at months 1–2, 3–4, 5–7, >7). The mRNA expression was measured by reverse transcription polymerase chain reaction (RT-qPCR). The mRNA expression levels were normalized to the geometric mean of all three constantly expressed reference genes (cyclophilin A, ubiquitin, ubiquitin C). Our findings suggest that adipokines are expressed and present in all investigated groups, and are specifically up- or downregulated during the estrus cycle and during pregnancy. Vaspin and adiponectin levels were upregulated in the middle and late cycle stages. Resistin was abundant during the CL regression stage and in the first months of pregnancy. The specific expression of adipokine receptors indicates their involvement in the local mechanisms that regulate CL function. Further investigations are required to elucidate the regulative mechanisms underlying the different local effects of adipokines on the ovarian physiology of cows.

The objective of the study was to evaluate the expression patterns of prostaglandin F2alpha (PGF), prostaglandin E2 (PGE), PGF receptor (FP), PGE receptors (EP2 and EP4), prostaglandin-endoperoxide synthase 2 (PTGS2) and prostaglandin synthases (PGFS and PGES) in corpora lutea (CL) during experimentally induced luteolysis in cow. The Fleckvieh cows in the mid-luteal phase (days 8-12, control group) were injected with cloprostenol (PGF analogue), and CL were collected by transvaginal ovariectomy before (days 8-12, control group) and at 0.5, 2, 4, 12, 24, 48 and 64 h after PGF application (n = 5 per group). The mRNA expression was determined by RT-qPCR, the hormone concentrations by enzyme immunoassay and localization by immunohistochemistry. PTGS2 gene expression increased significantly 2 h after PGF application, followed by continuous and significant downregulation afterwards. The PGF tissue concentration increased significantly just after PGF injection and again during structural luteolysis (after 12 h), whereas PGE concentration significantly decreased during structural luteolysis. The FP receptor mRNA decreased significantly at 2 h and again at 12 h after PGF. In contrast, EP4 receptor mRNA increased significantly just after the PGF application (0.5 h). The immunostaining of PGES and PTGS2 on day 15-17 shows numerous positive luteal cells, followed by lower activity afterwards on day 18 (luteolysis). In conclusion, the changes of examined prostaglandin family members in CL tissue after PGF application may be key components of the local mechanisms regulating the cascade of actions leading to functional and subsequent structural luteolysis in the bovine ovary.

Citrinin (CIT) is a mycotoxin responsible for the contamination of many agricultural products, like wheat, barley, corn, rice and their products, as also other foodstuffs and feedstuffs used in human and animal nutrition. It is essentially produced by Penicillium citrinum, although it can also be biosynthesised from Penicillium expansum and Penicillium verrucosum and some species of Aspergillus and Monascus. However, several studies have shown that CIT is known for its genotoxic, hepatotoxic, fetotoxic and teratogenic properties. The aim of this study is to investigate the occurrence of CIT in wheat grain cultivated in Kosovo andAlbania. Given the fact that wheat flour is the most consumed product in Kosovo and Albania, it is necessary to analyse the CIT in wheat in these two countries. In total, 60 wheat samples were tested from Fusha e Kosovës (Kosovo), Myzeqeja (Albania) and Fusha e Maliqit (Albania), as places withthe highest wheat production. The enzyme- linked immunosorbent assay (ELISA) method was used to determine CIT concentrations. To identify moulds representing potential producers of CIT, traditional macroscopic and microscopic methods and the molecular PCR method of identification were implemented. CIT was detected in 96.6% and 86.6% of wheat grain samples collected in Kosovo and Albania, respectively. The maximum amount of CIT detected in wheat grain was 53.12 μg/ kg in Kosovo, and 45.74 μg/kg in Albania. The amount of CIT found in wheat grain is not comparable with the maximal limits (MLs), as the European legislation does not provide limits for this mycotoxin. However, since there is generally a lack of data about CIT in cereals in Kosovo and Albania, the results can serve as an indicator of wheat grain contamination in this part of the Balkan Peninsula.

The objective of the study was to characterize expression patterns of hypoxia-inducible factor-1alpha and vascular endothelial growth factor isoforms during periovulation in the cow. Ovaries containing follicles or corpora lutea were collected by transvaginal ovariectomy (n =5 cows/group) and timely defined (after gonad otrop in-re leasi ng hormone application) as follows: (I) 0 h; (II) 4 h; (III) 10 h; (IV) 20 h; (V) 25 h (follicles); and (VI) 60 h (early corpora lutea). The mRNA expression of genes encoding hypoxia-inducible factor-1alpha and vascular endothelial growth factor isoforms was quantified by reverse transcriptase quantitative polymerase chain reaction. The high mRNA expression of hypoxia-inducible factor-1alpha in the control follicle group (before gonad otrop in-re leasi ng hormone) was followed by a conti-nuous and significant downregulation afterward with a minimum level in follicle group close to ovulation, and a significant increase just after ovulation. The mRNA of vascular endothelial growth factor isoforms was high during luteinizing hormone peak (4 h after gonad otrop in-re leasi ng hormone), decreased significantly thereafter to the lowest levels in follicle group 25 h after gonadotrop in-re leasi ng hormone, followed by a rapid and significant increase just after ovulation (newly corpora lutea). In conclusion, our results of hypoxia-inducible factor-1alpha and vascular endothelial growth factor regulation in follicle groups around ovulation indicate them to be necessary mediators of the luteinizing hormone-dependent ovulation and significant molecular mechanisms of angiogenesis during early corpora lutea formation.

Aflatoxin M-1 is a hydroxylated metabolite excreted in milk, which is considered as a potent health risk factor for consumers, therefore, the routine control of this toxin is essential. To monitor the concentration of this toxin, during this study are used and compared to each other for the correlation, two methods, the competitive method ELISA and UHPLC-ESI-MS/MS as a confirmative method. According to the results found using the two methods, from 192 of raw cow's milk samples analyzed, about 40% of samples resulted positive with AFM(1), among them, 6.5 % of the samples exceeded the maximum tolerable level according to ELISA method, and 5.5 % of the samples according to UHPLC-ESI-MS/MS. In conclusion, the results of this study suggest that there is a good correlation between the two methods used. The UHPLC-ESI-MS/MS method requires longer time of determination than ELISA method because there is the need of the extraction of milk samples for AFM(1) by the immunoaffinity columns ahead of quantitative analysis.

Food adulteration and fraudulent practices are widely observed in the food industry worldwide and are of great concern for Balkan countries. This study aims at investigating the level of undeclared pork meat in commercial beef and chicken meat products sold in Kosovo by implying one commercial enzyme-linked immunosorbent assay (ELISA) and two confirmatory real-time polymerase chain reaction (PCR) approaches [ready-to-use real-time PCR and real-time PCR with primers specific for pork mitochondrial deoxyribonucleic acid (DNA)]. In supermarkets in the capital city, Prishtina, 62 meat products were randomly sampled, and the three methods were applied. Additionally, these three approaches were evaluated for their practicability, reproducibility, and cost. The results showed that pork was present in 32% of beef- and 8% chicken-based products. ELISA and real-time PCR with pork specific primers showed 100% of reproducibility for beef- and chicken-based products. In contrast, the ready-to-use real-time PCR kit showed 100% reproducibility in chicken-, but only 75% in beef-based samples. ELISA was more rapid than both real-time PCR approaches, but it was more challenging when large numbers of samples were processed. The real-time PCR approach with pork specific primers was the cheapest, while the ready-to-use real-time PCR was the most practical method. Commercial ELISA, in combination with real-time PCR with pork specific primers, provides a reliable and affordable testing methodology that can be implemented for rapid detection and monitoring of pork adulteration in diverse commercial foods.

The objective of the study was to characterize expression patterns of hypoxia-inducible factor-1alpha (HIF1A), inducible nitric oxide synthase (iNOS) and endothelial (eNOS) isoforms in time-defined follicle classes before and after GnRH application in the cow. Ovaries containing preovulatory follicles or corpora lutea were collected by transvaginal ovariectomy (n=5 cows/group) as follow: (I) before GnRH administration; (II) 4h after GnRH; (III) 10h after GnRH; (IV) 20h after GnRH; (V) 25h after GnRH, and (VI) 60h after GnRH (early corpus luteum). The mRNA abundance of HIF1A in the follicle group before GnRH was high, followed by a significant down regulation afterwards with a minimum level 25h after GnRH (close to ovulation) and significant increase only after ovulation. The mRNA abundance of iNOS before GnRH was high, decreased significantly during LH surge, with minimum levels afterwards. In contrast, the mRNA of eNOS decreased in the follicle group 20h after GnRH, followed by a rapid and significant upregulation just after ovulation. Immunohistochemically, the granulosa cells of antral follicles and the eosinophils of the theca tissue as well of the early corpus luteum showed a strong staining for HIF1A. The location of the eosinophils could be clearly demonstrated by immunostaining with an eosinophil-specific antibody (EMBP) and transmission electron microscopy. In conclusion, the parallel and acute regulated expression patterns of HIF1A and NOS isoforms, specifically during the interval between the LH surge and ovulation, indicates that these paracrine factors are involved in the local mechanisms, regulating final follicle maturation, ovulation and early luteal angiogenesis.

Camaj, A., Arbneshi, T., Berisha, B., Haziri, A., & Camaj, A. (2019). Evaluation of afl atoxin M1 by ELISA in raw milk in Kosovo during 2016. Bulgarian Journal of Agricultural Science, 25(1), 124–128 Afl atoxins are a class of mycotoxins that are found in agricultural crops produced mainly by fungi Aspergillus fl avus and A. parasiticus. When afl atoxin B1 found in the food crops is consumed by cows, metabolizes to afl atoxin M1 (AFM1), which is excreted through milk. Afl atoxin M1 is a human toxin that causes several health problems. In 2016, during the winter and the summer season, 192 raw cow milk samples were collected from small farms in fi ve major regions of Kosovo (Prishtina, Gjilan, Mitrovica, Peja, and Prizren), and analyzed for the afl atoxin M1 level, using Enzyme-Linked Immunosorbent Assay (ELISA). The number of positive samples was 80 (41.7%), from which 43 samples were from the winter season and 37 samples from the summer season. Prizren was the region with the highest number of positive samples for the two seasons examined. The region with the highest percentage of samples that exceeded the maximum tolerable limit for AFM1, according to EU Maximum Tolerable Limit, was Prishtina, with 15.4% of the samples. The difference of AFM1 level between the seasons was statistically signifi cant (P ≤ 0.05 to P ≤ 0.001). The maximum mean concentrations of AFM1 recorded in the winter season were in the range of 0.004-0.109 ng/mL, and during the summer season they are in the range of 0.004-0.833 ng/mL, all coming from the region of Gjilan. The maximum range for this region resulted to be alarming, values that needed to be reevaluated by a confi rming method, like HPLC, in another study. During 2016, the occurrence of AFM1 in raw cow’s milk appears to be prevalent in Kosovo.

Total mesophilic bacteria, anaerobic bacteria, lactic acid bacteria, enterobacteria as well as yeast and moulds were investigated during the production of the traditional Rugova cheese without using any starter culture. This type of cheese has been produced and consumed by the local population in the Rugova region for centuries, and is important for their economy. Samples from different stages of production (raw milk, curd and cheese during maturation on day 1, 8 and 14) were collected from local farms in the Rugova region, Kosovo, during winter. After sample collection and their serial dilution, samples were plate on different media and incubated at the appropriate temperature. According to identical morphology and physiology the number of 52 isolates obtained from de Man, Rogosa, and Shape (MRS), and M 17 agar was reduced to ten Gram-positive and catalase negative strains. These strains were subjected to API 50 CHL fermentation profiling. There was a variety of microbial groups detected in all stages of cheese production, with the dominant microbial groups being Lactococcus (from 6.44 log cfu/ml in milk to 8.39 log cfu/gr in cheese D14) and Lactobacillus (from 5.79 log cfu/ml in milk to 7.58 log cfu/gr in cheese D14). Enterobacter and yeast and moulds were found in lower amounts compared to the other microbes in all stages. The level of Leuconostoc was approximately the same as Lactobacillus. Almost all microbial groups were low in curd and started to increase from day one of cheese manufacturing. Three of the ten isolates were identified as Lactococcus lactis ssp. lactis, four as Lactobacillus plantarum (2 isolates) and L. curvatus (2 isolates). One isolate of each of Lactobacillus paracasei ssp. paracasei, L. brevis and L. pentosus were identified. These results confirm the competitiveness of high numbers of LAB during fermentation of this cheese.

Aflatoxins are a class of mycotoxins that are found in agricultural crops produced mainly by fungi Aspergillus flavus and A. parasiticus. When aflatoxin B1 found in the food crops is consumed by cows, metabolizes to aflatoxin M1 (AFM1), which is excreted through milk. Aflatoxin M1 is a human toxin that causes several health problems. In 2016, during the winter and the summer season, 192 raw cow milk samples were collected from small farms in five major regions of Kosovo (Prishtina, Gjilan, Mitrovica, Peja, and Prizren), and analyzed for the aflatoxin M1 level, using Enzyme-Linked Immunosorbent Assay (ELISA). The number of positive samples was 80 (41.7%), from which 43 samples were from the winter season and 37 samples from the summer season. Prizren was the region with the highest number of positive samples for the two seasons examined. The region with the highest percentage of samples that exceeded the maximum tolerable limit for AFM1, according to EU Maximum Tolerable Limit, was Prishtina, with 15.4% of the samples. The difference of AFM1 level between the seasons was statistically significant (P <= 0.05 to P <= 0.001). The maximum mean concentrations of AFM1 recorded in the winter season were in the range of 0.004-0.109 ng/mL, and during the summer season they are in the range of 0.004-0.833 ng/mL, all coming from the region of Gjilan. The maximum range for this region resulted to be alarming, values that needed to be reevaluated by a confirming method, like HPLC, in another study. During 2016, the occurrence of AFM1 in raw cow's milk appears to be prevalent in Kosovo.