Complete genome of Microbacterium plantarum CoE-159-22, isolated from traditionally produced Montenegrin cheese.

Microbacterium plantarum (M. plantarum) was recently described as a new species isolated from copper globemallow (Sphaeralcea angustifolia). Here, we report the complete genome of M. plantarum CoE-159-22, which was obtained from traditionally produced Montenegrin cheese.

Cadmium and lead in fresh vegetables and vegetable products sold on the local market in the Republic of Serbia, 2018-2021.

Cadmium (Cd) and lead (Pb) concentrations were determined in samples of 31 species of vegetables (n = 719) and in vegetable products (n = 17), collected during the period January 2018 to September 2021. These originated from 33 countries, including Serbia. The samples were analysed by inductively coupled plasma - optical emission spectrometry (ICP-OES). Overall, Cd and Pb were found above the limit of detection in 123 (16.7%, n = 736) and 90 (12.2%; n = 736) samples, respectively. According to the former legislation, valid until the end of August 2021, the maximum levels (MLs) of Cd and Pb were exceeded in 1 and 2 samples of vegetables, respectively. Regarding the EU and Serbian legislation which is valid since September 2021 the MLs of Cd and Pb for vegetables were exceeded in 7 samples: Cd in 5 and Pb in 2 samples. In addition, 3 vegetable product samples exceeded the MLs for both cadmium and lead according to both former and current regulations.

Enterococcus montenegrensis sp. nov., isolated from artisanal Montenegrin dry sausage.

A novel, Gram-positive, facultative anaerobe, coccoid and non-motile bacterium, designated as CoE-012-22T was isolated from dried beef sausage (the original name in Montenegro is Govedji Kulen) manufactured in the municipality of Rozaje (Montenegro) in 2021. Cells of this strain were oxidase- and catalase-negative. Growth occurred at 4-50 °C, at pH 5.0-8.0 and with 0-6.5 % (w/v) NaCl in diverse growth media. MALDI-TOF analysis identified the strain as Enterococcus canintestini (log score 2). Phylogenetic analysis of the 16S rRNA gene and whole genome sequences assigned the strain to the genus Enterococcus. The closest relatives were E. canintestini DSM 21207T and E. dispar ATCC 51266T with 16S rRNA gene sequence pairwise similarities of 99.34 and 98.59 %, respectively. The average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) values between isolate CoE-012-22T and other enterococci species were below the thresholds for species delineation thresholds (95.0 % ANI; 70.0 % dDDH) with maximum identities of 84.13 % (ANIb), 86.43 % (ANIm) and 28.4 % (dDDH) to E. saigonensis JCM 31193T and 70.97 % (ANIb), 88.99 % (ANIm) and 32.4 % (dDDH) to E. malodoratus ATCC 43197T. Two unknown Enterococcus isolates, Enterococcus sp. MJM12 and Enterococcus SMC-9, showed identities of 99.87 and 99.94 % (16S rRNA), 98.57 and 98.65 % (ANIb), 98.93 and 99.02 % (ANIm), and 89.8 and 90.0 % (dDDH) to strain CoE-012-22T and can therefore be regarded as the same species. Based on the characterization results, strain CoE-012-22T was considered to represent a novel species, for which the name Enterococcus montenegrensis sp. nov. is proposed. The type strain is CoE-012-22T (=DSM 115843T=NCIMB 15468T).

Cadmium and lead in fresh fruits and fruit products sold on the local market in the Republic of Serbia, 2018-2021.

Cadmium (Cd) and lead (Pb) concentrations were determined in 737 samples of 39 species of fruits and of processed fruits (n = 13), collected during the period January 2018 to September 2021. Fruits and fruit products originated from 47 countries, including Serbia. Samples were analysed by inductively coupled plasma - optical emission spectrometry (ICP-OES). Overall, Cd and Pb were found above the limit of detection (LOD) in 97 samples (13.9%). Cd and Pb concentrations were below the LOD in all samples of fruit products. According to the valid regulations before September 2021, the maximum level (ML) of Cd for analysed fruits was exceeded in only 1 sample (0.14%; n = 737), while concentrations of Pb in each analysed fruit sample was below the ML. According to the legislative regulations as set after September 2021, the maximum levels of Cd and Pb for fruits were exceeded in 5, respectively 1, of the samples, so 6 in total (0.81%; n = 737).

The association of socio-demographic characteristics of university students and the levels of their digital entrepreneurial competences.

The aim of this article is to provide empirical evidence on the level of digital entrepreneurial competences (DEC) that higher education students acquire during their higher education and to investigate the relationship between their socio-demographic characteristics and the level of DEC. The first competence area (Identification of opportunities) of the DEC EmDigital competence framework was assessed together with its three sub-competence areas (Search for and analysis of information, Creativity and innovation, and Prospecting), and the relationship between the level of DEC and various socio-demographic characteristics of higher education students was investigated using logistic regression. The results show a relatively low DEC level of the participating students for the first EmDigital competence area. Location, field of study, level of study and employment status were found to be statistically significant for the acquisition of DEC, while the variables age and gender had little impact. The results of this study are a step towards the development of an DEC online assessment tool (using the Digital Competence Wheel as a role model) that universities and other stakeholders could use to assess the level of DEC their students acquire during university education. This research is the first study to assess the acquisition of DEC during university studies and the impact of different socio-demographic characteristics of students on their level of DEC.

Cadmium, lead, mercury and arsenic in fresh vegetables and vegetable products intended for human consumption in the Republic of Serbia, 2015-2017.

The concentrations of cadmium (Cd), lead (Pb), mercury (Hg) and arsenic (As) were determined in 455 samples of 27 species of vegetables and 28 different processed vegetables collected during the period from January 2015 to December 2017. Vegetables (n = 387) and vegetable products (n = 68) originated from 31 countries, including Serbia. The samples were analysed by inductively coupled plasma - optical emission spectrometry (ICP-OES). The concentrations of Cd, Pb, Hg and As in the vegetables and vegetable products were compared to the maximum levels set by the European Union and the Serbian legislation. The concentration of mercury was less than the limit of detection in each analysed sample. One or multiple measurable toxic metals (Cd, Pb and/or As) were found in 250 samples (54.9%; n = 455). According to the Regulations which were valid until the end of August 2021, the maximum levels of Cd, Pb and As were exceeded in 19 samples (4.2% of the samples of vegetable and vegetable products; n = 455), i.e. in 13 samples of vegetables: Cd in three, Pb in nine and As in one sample and in 6 samples of vegetables products: Cd in three, Pb in one and As in two samples. Regarding the new EU and Serbian legislation which is valid since September 2021 the maximum levels of Cd and Pb for vegetables and vegetable products were exceeded in 118 samples (25.9% of the samples of vegetable and vegetable products; n = 455), i.e. in 95 samples of vegetables: Cd in 67 and Pb in 28 samples and in 23 samples of vegetable products: Cd in 20 and Pb in 3 samples.

Draft Genome Sequence of Enterococcus dispar CoE-457-22, Isolated from Traditionally Produced Montenegrin Dry Sausage.

Enterococcus dispar was isolated for the first time from synovial fluid and stool cultures and described as a new species in 1991. Here, we report the genome of E. dispar CoE-457-22, which was obtained from traditionally produced Montenegrin dry sausage (sudzuk).

Cadmium, lead, mercury, and arsenic in fresh fruits and fruit products intended for human consumption in the Republic of Serbia, 2015-2017.

The European Commission has established legislation for toxic metals in certain foodstuffs in order to protect public health already in 1993. After several amendments, new maximum levels for cadmium and lead in certain foods were set as per 30 and 31 August 2021 for lead and cadmium, respectively. The new Serbian Regulation on "maximum levels of certain contaminants in food" is fully harmonised with this European legislation. The concentrations of cadmium (Cd), lead (Pb), mercury (Hg), and arsenic (As) were determined in 832 samples of 39 species of fruits and processed fruits, collected during the period January 2015 to December 2017. Fruits and fruit products originated from 45 countries, including Serbia. Samples were analysed by inductively coupled plasma - optical emission spectrometry (ICP-OES). Concentrations of cadmium, lead, mercury, and arsenic in fruits and fruit products were compared to these maximum levels (MLs). Overall, measurable Cd, Pb, Hg, and As were found in 377 samples (45.3%). According to the former regulations, the maximum levels of Cd and Pb for analysed fruits and fruit products were exceeded in only 10 samples (1.20%; n = 832): Cd in 6 and Pb in 4 samples, but according to the latest regulations, the maximum levels of Cd and Pb for fruits were exceeded in 196 of the samples (23.9%; n = 820): Cd in 191 and Pb in 5 samples. The increase of ML exceedances shows that attention needs to be paid for compliance of food to the recent MLs for these metals.

Genome Sequences of Lactococcus garvieae and Lactococcus petauri Strains Isolated from Traditional Montenegrin Brine Cheeses.

Lactococcus garvieae and Lactococcus petauri cause lactococcosis in fish. Both species have also been isolated from various food products and are considered emerging zoonotic pathogens. Here, we report the genomes of L. garvieae INF126 and L. petauri INF110, obtained from traditional Montenegrin brine cheeses.

Genetic Diversity of Leuconostoc mesenteroides Isolates from Traditional Montenegrin Brine Cheese.

In many dairy products, Leuconostoc spp. is a natural part of non-starter lactic acid bacteria (NSLAB) accounting for flavor development. However, data on the genomic diversity of Leuconostoc spp. isolates obtained from cheese are still scarce. The focus of this study was the genomic characterization of Leuconostoc spp. obtained from different traditional Montenegrin brine cheeses with the aim to explore their diversity and provide genetic information as a basis for the selection of strains for future cheese production. In 2019, sixteen Leuconostoc spp. isolates were obtained from white brine cheeses from nine different producers located in three municipalities in the northern region of Montenegro. All isolates were identified as Ln. mesenteroides. Classical multilocus sequence tying (MLST) and core genome (cg) MLST revealed a high diversity of the Montenegrin Ln. mesenteroides cheese isolates. All isolates carried genes of the bacteriocin biosynthetic gene clusters, eight out of 16 strains carried the citCDEFG operon, 14 carried butA, and all 16 isolates carried alsS and ilv, genes involved in forming important aromas and flavor compounds. Safety evaluation indicated that isolates carried no pathogenic factors and no virulence factors. In conclusion, Ln. mesenteroides isolates from Montenegrin traditional cheeses displayed a high genetic diversity and were unrelated to strains deposited in GenBank.

Effect of Tympanostomy Tube Placement on Intraoperative Auditory Brainstem Response.

BACKGROUND: Intraoperative auditory brainstem response (ioABR) testing under general anesthesia is commonly performed on children when sleep-deprived ABR and behavioral testing are not reliable or feasible. Several studies have reported potential confounding results when tube insertion is combined with ABR testing. PURPOSE: This article evaluates whether a temporary threshold shift (TTS) occurs following placement of tympanostomy tubes (TTs) in children who undergo ioABR testing. RESEARCH DESIGN: A case-control prospective study. Patients scheduled for combined TT and ioABR procedures were enrolled into this study. STUDY SAMPLE: Twenty children (38 ears), ranging in age from 6 months to 10.5 years, were enrolled. DATA COLLECTION AND ANALYSIS: ABR thresholds for tone bursts with center frequencies of 2 and 4 kHz were compared before and after tube insertion. The indication for surgery, comorbidities, and status of the middle ear were also recorded. A paired t-test was used to determine statistical significance. RESULTS: Data collection did not necessarily indicate suctioning and tube placement were causing a TTS; however, fluid present in the middle ear space prior to TT placement appeared to influence results. Although a paired t-test did not show statistically significant differences in ABR thresholds between groups of individuals with and without fluid and before and after tube placement, 50% of patients with mucoid fluid (two out of four patients) were noted to have 10 dB or greater worsening in ABR thresholds in two out of six ears. Twenty-nine percent of patients with serous fluid (two out of seven patients) were noted to have at least a 10 dB worsening in ABR threshold in 2 out of 10 ears. Further testing is needed to confirm these trends. CONCLUSION: This pilot study indicates that children with mucoid or serous fluid may experience worse ABR thresholds following TT insertion. ioABR testing immediately after TT tube placement and in the presence of middle ear fluid should be interpreted with caution. Additional studies with a larger sample size are needed to confirm these results.

New Formulation towards Healthier Meat Products: Juniperus communis L. Essential Oil as Alternative for Sodium Nitrite in Dry Fermented Sausages.

The effect of Juniperus communis L. essential oil (JEO) addition at concentrations of 0.01, 0.05 and 0.10 µL/g on pH, instrumental parameters of color, lipid oxidation (2-Thiobarbituric acid reactive substances (TBARS)), microbial growth, texture and sensory attributes of dry fermented sausages produced with different levels of fat (15 and 25%) and sodium nitrite (0, 75 and 150 mg/kg) was assessed. Reduced level of sodium nitrite (75 mg/kg) in combination with all three concentrations of JEO (0.01-0.10 µL/g) resulted in satisfying physico-chemical (color and texture) properties and improved oxidative stability (TBARS < 0.3 mg MDA/kg) of dry fermented sausages produced with 25% of fat. However, sausages produced with 0.10 µL/g of JEO had untypical flavor. No foodborne pathogens (Escherichia coli, Listeria monocytogenes, Salmonella spp. and sulfite-reducing clostridia) were detected in any sample throughout the storage period (225 days). The results of this study revealed significant antioxidative activity of JEO and consequently its high potential as effective partial replacement for sodium nitrite in dry fermented sausages.

Role of cochlear synaptopathy in cytomegalovirus infected mice and in children.

OBJECTIVES: Determine whether a murine model of cytomegalovirus (CMV) and CMV- infected children show evidence of synaptopathy. STUDY DESIGN: Murine model of CMV infection and case series. SUBJECTS AND METHODS: C57 BL/6 mice were inoculated with murine-CMV (mCMV). Auditory function was assessed using Auditory Brainstem Response (ABR) and distortion product otoacoustic emission (DPOAE) testing. Temporal bones from mCMV-infected mice were used for both ribbon synapse and hair cell quantification. Four groups of children (non-CMV normal hearing, non-CMV hearing impaired, CMV normal hearing and CMV hearing impaired) underwent ABRs between 2014 and 2018. The outcomes included raw amplitude, wave I:V amplitude ratio, absolute latency, and interpeak latency. RESULTS: Mice at 8 weeks post mCMV infection had higher ABR and DPOAE (P < 0.05) thresholds and increased outer hair cell loss compared to uninfected mice and mCMV-infected mice at 4 and 6 weeks post infection, indicating progressive hearing loss. A reduction in the wave I amplitude and synaptic counts were noted earlier at 4 weeks in CMV-infected mice (P < 0.05). The human data indicated that the wave I:V amplitude ratio was lower on average in CMV-infected groups when compared to the uninfected cohorts. The wave I:V amplitude ratio for the click and 4k stimuli were not significantly different between the congenital CMV-infected and uninfected children with normal or with hearing loss. CONCLUSION: This study suggests mCMV infection results in a synaptopathy before hair cell damage. Additional studies need to be performed to determine whether this effect is also observed in CMV-infected children. LEVEL OF EVIDENCE: Animal studies and basic science- NA; human studies: level 4.

Draft Genome Sequences of Five Enterococcus faecium Isolates from Traditional Montenegrin Brine Cheese.

Enterococcus faecium is a multifaceted bacterial species. It is part of the natural human microbiota, it grows in a variety of traditional foods, and emerging multiresistant clones are a leading cause of nosocomial infections. Here, we present draft genomes of five E. faecium isolates originating from traditional Montenegrin brine cheeses.

Lactococci of Local Origin as Potential Starter Culturesfor Traditional Montenegrin Cheese Production.

The aim of this study is to characterise and examine the biochemical properties of 40 Lactococcus lactis strains isolated from indigenous Montenegrin dairy products in order to explore their potential to be used as starter cultures for producing typical Montenegrin cheese, such as 'bijeli sir', 'masni sir' and 'njeguski sir'. Their safety regarding the production of biogenic amines, the presence of antimicrobial resistance and the antibacterial activity against relevant pathogens and spoilage microorganisms has also been tested. Based on the characterisation, all strains belong to L. lactis ssp. lactis. Out of these 40 strains, 23 displayed rapid acidification ability and proteolysis. However, none of the strains exhibited the ability of lipid degradation. Most of the strains were not associated with any health risk investigated. Summing up, a large percentage (27.5%) of the tested strains showed good properties. These strains should be further examined for their possible application as specific starter cultures in the production of indigenous cheese in Montenegro.

Cadmium in liver and kidneys of domestic Balkan and Alpine dairy goat breeds from Montenegro and Serbia.

Concentrations of cadmium (Cd) were determined in the samples of 144 animals around 1 and of 144 animals around 4 years old. Cd was analysed by inductively coupled plasma-optical emission spectrometry (ICP-OES), after microwave digestion. Cd concentrations were higher (p < 0.05) in kidney than in liver and higher (p < 0.05) in older animals than in young ones. In domestic Balkan goat which was raised in a free-ranged system Cd accumulation was lower (p < 0.05) than in Alpine goat raised in an intensive production system. Geographic region did influence Cd accumulation only in older animals. Higher Cd levels (p < 0.05) were determined in goats from Serbia. The highest obtained Cd concentrations in both tissues were lower than maximum levels set by European and national legislation for ruminants (cattle and sheep).

Cadmium and lead in female cattle livers and kidneys from Vojvodina, northern Serbia.

Concentrations of cadmium (Cd) and lead (Pb) were determined in livers (n = 52) and kidneys (n = 52) of female cattle (345-2717 days old) from dairy farms in the region Vojvodina. Cd and Pb were analysed by inductively coupled plasma-optical emission spectrometry, after microwave digestion. Cd and Pb concentrations did not exceed the Serbian and European maximum set limits in any sample. The Cd concentrations in the livers and kidneys ranged from 0.033 to 0.151 mg kg-1 wet weight and from 0.055 to 0.510 mg kg-1 wet weight, respectively. The corresponding Pb concentrations were 0.015-0.159 mg kg-1 wet weight and 0.021-0.196 mg kg-1 wet weight, respectively. Mean Cd and Pb concentrations were significantly lower (p < 0.001) in the liver (0.072 and 0.053 mg kg-1 wet weight) than in the kidney (0.190 and 0.075 mg kg-1 wet weight). There were good correlations between Cd in liver and Cd in kidney, Pb in liver and Pb in kidney, Cd level and age and Pb level and age in both tissues.